Friday, February 13, 2009
Saturday, February 7, 2009
SHATAVARI
SHATAVARI
Shatavari,
In India, Shatavari is considered the women's equivalent to Ashwagandha. The name translates to "she who possesses 100 husbands", referring to the herbs rejuvanitive effect upon the female reproductive organs. In Australia the herb is more often used to treat gastrointestinal disorders and as an external wash for wounds.
The healing qualities of Shatavari are useful to a wide array of ailments. It is well known for it effects on the female reproductive system. It is also effective in a number of other systems of the body and is therefore of use to both men and women.
Botanical Name : Asparagus racemosus
Sanskrit -- Shatavari
Hindi -- Shatavari
English -- Indian Asparagus, Hundred Roots , Asparagus roots
Chinese - Tian men dong
Family : Asparagaceae
Effect on the Doshas: Vata: - Pitta: - Kapha: +
Rasa (Taste) : Madhura (sweet) ,Tikta (bitter)
Virya (Energy): shita (cold)
Vipak (Post-Digestive Action): madhura
Parts Used : Roots (Rhizomes) and leaves.
Properties :
o Nutritive tonic, rejuvenative,
o aphrodisiac,
o galactogogue
o laxative
o antispasmodic,
o Antacid
o diuretic
o antitumor
o demulcent
Indications:
Shatavari is perhaps best known as a female rejuvanitive. It is useful for infertility, decreased libido, threatened miscarriage, menopause, leucorrhea and has the ability to balance pH in the cervical area. Dry membranes, such as those on the vaginal wall, are also brought into balance through the herbs demulcent action.
Men may benefit from the herb as well in the treatment of impotence and general sexual debility. In addition to it's applications for reproductive organs, Shatavari is also quite effective for stomach ulcers, hyperacidity and diarrhea. Dry and irritated membranes in the upper respiratory tract are soothed by this herb making it useful in cases of bronchitis and chronic fevers. It is believed to bring into balance all of the body's fluids.
Medicinal Uses:
The most important herb in Ayurvedic medicine for women
Used internally for infertility, loss of libido, threatened miscarriage, menopausal problems. It both nourishes and cleanses the blood and the female reproductive organs. It is a good food for menopause or for those who have had hysterectomies, as it supplies many female hormones. It nourishes the ovum and increases fertility. This herb is known to increase Sattva, or positivity and healing power. It also enhances the feelings of spiritual love, and increases Ojas.
The male reproductive system will also benefit from Shatavari. It may be used in cases of sexual debility, impotence, spermatorrhea, and inflammation of sexual organs.
Useful for hyperacidity, stomach ulcers, dysentery, and bronchial infections.
Contraindications (Cautions): massive fibrocystic breasts, estrogen induced problems, estrogen induced fibrocystic changes or other problems
Chemical constituents :
Shatavari is rich in active constituents such as galactose, arabinose, steroidal glycosides and saponins.
arabinose
Shatavari,
Asparagus racemosus, is a climbing plant which grows in low jungles areas throughout India. This sweet and bitter herb is particularly balancing to Pitta Dosha.
In India, Shatavari is considered the women's equivalent to Ashwagandha. The name translates to "she who possesses 100 husbands", referring to the herbs rejuvanitive effect upon the female reproductive organs. In Australia the herb is more often used to treat gastrointestinal disorders and as an external wash for wounds.
The healing qualities of Shatavari are useful to a wide array of ailments. It is well known for it effects on the female reproductive system. It is also effective in a number of other systems of the body and is therefore of use to both men and women.
Botanical Name : Asparagus racemosus
Sanskrit -- Shatavari
Hindi -- Shatavari
English -- Indian Asparagus, Hundred Roots , Asparagus roots
Chinese - Tian men dong
Family : Asparagaceae
Effect on the Doshas: Vata: - Pitta: - Kapha: +
Rasa (Taste) : Madhura (sweet) ,Tikta (bitter)
Virya (Energy): shita (cold)
Vipak (Post-Digestive Action): madhura
Parts Used : Roots (Rhizomes) and leaves.
Properties :
o Nutritive tonic, rejuvenative,
o aphrodisiac,
o galactogogue
o laxative
o antispasmodic,
o Antacid
o diuretic
o antitumor
o demulcent
Indications:
Shatavari is perhaps best known as a female rejuvanitive. It is useful for infertility, decreased libido, threatened miscarriage, menopause, leucorrhea and has the ability to balance pH in the cervical area. Dry membranes, such as those on the vaginal wall, are also brought into balance through the herbs demulcent action.
Men may benefit from the herb as well in the treatment of impotence and general sexual debility. In addition to it's applications for reproductive organs, Shatavari is also quite effective for stomach ulcers, hyperacidity and diarrhea. Dry and irritated membranes in the upper respiratory tract are soothed by this herb making it useful in cases of bronchitis and chronic fevers. It is believed to bring into balance all of the body's fluids.
Medicinal Uses:
The most important herb in Ayurvedic medicine for women
Used internally for infertility, loss of libido, threatened miscarriage, menopausal problems. It both nourishes and cleanses the blood and the female reproductive organs. It is a good food for menopause or for those who have had hysterectomies, as it supplies many female hormones. It nourishes the ovum and increases fertility. This herb is known to increase Sattva, or positivity and healing power. It also enhances the feelings of spiritual love, and increases Ojas.
The male reproductive system will also benefit from Shatavari. It may be used in cases of sexual debility, impotence, spermatorrhea, and inflammation of sexual organs.
Useful for hyperacidity, stomach ulcers, dysentery, and bronchial infections.
Contraindications (Cautions): massive fibrocystic breasts, estrogen induced problems, estrogen induced fibrocystic changes or other problems
Chemical constituents :
Shatavari is rich in active constituents such as galactose, arabinose, steroidal glycosides and saponins.
arabinose
Photochemical Screening of Crude Drugs
Photochemical Screening of Crude Drugs
S.NO TEST NAME TEST OBSERVATION
1. ALKOLOIDS
A .Mayer’s Test Sample + Pot. Mercuric Iodide sol Cream colour ppt
B . Dragondorff’s Sampl + Pot. Bismuth Iodide sol Reddish brown ppt
C . Wagner’s Sample + I2 in KI “
D. Hager’s Sample + saturated picric acid Yellow colour ppt
2. CARBOHYDRATES
A . Molisch’s Test Sample + α- napthol + conc. H2SO4 Purple colour
B. Fehling’s test Sample + [Fehling A+ B] , heat Brick red colour
3. GLYCOSUDES
A . Legal’s Test Extract + pyridine + sod.nitro preside + alkali Pink or red colour
B. Bortranger’s Drug is powered and further extracted with ether or any solvent. The filtered ethereal extract is made alkaline either with caustic soda or ammonia by which aq. Layer shows after shaking Pink or red or violet colour occurs
C. libermann’s CHCL3sol of sample + conc. H2SO4 Green colour
4. PHYTO STEROLS
A . libermann Chloroform sol of sterol + conc. H2SO4 +
Acetic anhydride Green colour
5. PROTEINS & A.A
A . MIllon’s Test MIllon’s Reagent (mercuric nitrate in HNO3)
Millon’s reagent + protein sol White ppt which turns red on heating
B. Biuret Test Alk. Sol of protein + dil. Sol of CuSO4. On heating Red or violet colour
C. Ninhydrin Test Protein sol + aq . ninhydrin. On heating Purple colour
6. GUMS & MUCILAGE
A . Molisch’s Test Sample + α- napthol + conc. H2SO4 Purple colour
B. Precipitation Test 1- lead acetate + sample Ppt is formed
2- sample + CaCl2 Copious ppt
S.NO TEST NAME TEST OBSERVATION
1. ALKOLOIDS
A .Mayer’s Test Sample + Pot. Mercuric Iodide sol Cream colour ppt
B . Dragondorff’s Sampl + Pot. Bismuth Iodide sol Reddish brown ppt
C . Wagner’s Sample + I2 in KI “
D. Hager’s Sample + saturated picric acid Yellow colour ppt
2. CARBOHYDRATES
A . Molisch’s Test Sample + α- napthol + conc. H2SO4 Purple colour
B. Fehling’s test Sample + [Fehling A+ B] , heat Brick red colour
3. GLYCOSUDES
A . Legal’s Test Extract + pyridine + sod.nitro preside + alkali Pink or red colour
B. Bortranger’s Drug is powered and further extracted with ether or any solvent. The filtered ethereal extract is made alkaline either with caustic soda or ammonia by which aq. Layer shows after shaking Pink or red or violet colour occurs
C. libermann’s CHCL3sol of sample + conc. H2SO4 Green colour
4. PHYTO STEROLS
A . libermann Chloroform sol of sterol + conc. H2SO4 +
Acetic anhydride Green colour
5. PROTEINS & A.A
A . MIllon’s Test MIllon’s Reagent (mercuric nitrate in HNO3)
Millon’s reagent + protein sol White ppt which turns red on heating
B. Biuret Test Alk. Sol of protein + dil. Sol of CuSO4. On heating Red or violet colour
C. Ninhydrin Test Protein sol + aq . ninhydrin. On heating Purple colour
6. GUMS & MUCILAGE
A . Molisch’s Test Sample + α- napthol + conc. H2SO4 Purple colour
B. Precipitation Test 1- lead acetate + sample Ppt is formed
2- sample + CaCl2 Copious ppt
Tylophora Indica
Tylophora Indica ( T. Asthmatica)
• Botanical Name : Tylophora Indica ( T. Asthmatica)
• Family Name : Asclepiadaceae
• Common Name : Emetic Swallow-wory, Indian Ipecac
• Part Used : Leaves, Roots
• Habitat : Throughout india, found in the plains, forests, hilly slopes and outskirts of the forest upto 900m.
• Product offered : Roots
Uses : The leaves and roots have emetic, cathartic, laxative, expectorant, diaphoretic and purgative properties. It has also been used for the treatment of allergies, cold, dysentery, hay fever and arthritis. It has reputation as an alterative and as a blood purifier, often used in rheumatism and syphilitic rheumatism. Root or leaf powder is used in diarrhoea, dysentery and intermittent fever. Dried leaves are emetic diaphoretic and expectorant. It is regarded as one of the best indigenous substitute for ipecacuanha. The roots are suggested to be a good natural preservative of food.
It is traditionally used as a folk remedy in certain regions of India for the treatment of bronchial asthma , inflammation , bronchitis, allergies, rheumatism and dermatitis. It also seems to be a good remedy in traditional medicine as anti-psoriasis, seborrheic, anaphylactic, leucopenia and as an inhibitor of the Schultz-Dale reaction. The roots are suggested to be a good natural preservative of food.
Active constituents
The major constituent in Tylophora is the alkaloid tylophorine. Laboratory research has shown this isolated plant extract exerts a strong anti-inflammatory action. Test tube studies suggest that tylophorine is able to interfere with the action of mast cells, which are key components in the process of inflammation. These actions seem to support Tylophoras traditional use as an anti - asthmatic and anti -allergenic medication by Ayurvedic
• Botanical Name : Tylophora Indica ( T. Asthmatica)
• Family Name : Asclepiadaceae
• Common Name : Emetic Swallow-wory, Indian Ipecac
• Part Used : Leaves, Roots
• Habitat : Throughout india, found in the plains, forests, hilly slopes and outskirts of the forest upto 900m.
• Product offered : Roots
Uses : The leaves and roots have emetic, cathartic, laxative, expectorant, diaphoretic and purgative properties. It has also been used for the treatment of allergies, cold, dysentery, hay fever and arthritis. It has reputation as an alterative and as a blood purifier, often used in rheumatism and syphilitic rheumatism. Root or leaf powder is used in diarrhoea, dysentery and intermittent fever. Dried leaves are emetic diaphoretic and expectorant. It is regarded as one of the best indigenous substitute for ipecacuanha. The roots are suggested to be a good natural preservative of food.
It is traditionally used as a folk remedy in certain regions of India for the treatment of bronchial asthma , inflammation , bronchitis, allergies, rheumatism and dermatitis. It also seems to be a good remedy in traditional medicine as anti-psoriasis, seborrheic, anaphylactic, leucopenia and as an inhibitor of the Schultz-Dale reaction. The roots are suggested to be a good natural preservative of food.
Active constituents
The major constituent in Tylophora is the alkaloid tylophorine. Laboratory research has shown this isolated plant extract exerts a strong anti-inflammatory action. Test tube studies suggest that tylophorine is able to interfere with the action of mast cells, which are key components in the process of inflammation. These actions seem to support Tylophoras traditional use as an anti - asthmatic and anti -allergenic medication by Ayurvedic
Kali musli
Kali musli
Botanical name: Curculigo orchioides
Family: Liliaceae (Lily family) (Hypoxidaceae)
Common name: Orchid palm grass • Hindi: Kali musli • Oriya: Tala-muli • Kannada: Nela tengu • M General Description: Kali Musli grows as Forest herb. Since generations, it is in use as folk medicine. In many parts of India, due to its over exploitation, Kali Musli is becoming rare in occurrence.
Botanical Description: Kali Musli is a herbaceous tuberous perennial with a short or elongate root stock bearing several fleshy lateral roots; Leaves sessile or petiolate 15-45x1.3-2.5 cm, linear or linear lanceolate, tips sometimes rooting, scape very short, clavate; Flowers in racemes, distichious, yellow, lowest in the racemes 2 sexual, perianth segments elliptic, oblong, hairy on the back; Fruits capsules, derived from inferior tricarpellary syncarpous ovary, 1-4 seeded; Seeds black, oblong, deeply grooved in wavy lines.
Useful Parts: All parts.
Habitat
It is found in places that are up to 6000 feet above sea level. It is more commonly seen in temperate zonal climate. It is seen in countries like Africa, Middle East, Arabia, Pakistan and southern Asian Island. In India it is found every where but predominantly found in Deccan plateau area of Chota Nagpur. It is also commonly seen in south India.
Morphology
It is a perennial shrub having short or elongated fleshy roots. Leaves are 6 to 18 inch in length and ½ to 1 ½ inch broad. It is speared shaped and bears stripes on it. The apex of the leaves is rooting. Flowers present the racemes presentation that is 1 inch in length. Flowers are ½ to 2/3 inch in diameter, shiny and are just above the ground. . Fruit is ½ inch in length, capsulated, ovate in shape, sharp at the apex and contains 1 to 4 seeds in it. Seeds are shiny, oblong in shape, 1/8 inch long, striped and sharp at the apex and base. Rhizome is 1 feet in length and is pulpy. The plant flowers in summers
Chemical Constituents
It contains starch 43.48 %, tannins 4.15 %, enzymes 14.18 % and ash 8.6 %. Besides these it also contains glycoside, orcinol-1-O-beta-D-apiofuranosyl-(1-->6)-beta-D-glucopyranoside, curculigoside, syringic acid, curculigoside and curculigoside C
Medicinal uses:
The rhizomes of the plants are used for the treatment of decline in strength, jaundice and asthma. According to Ayurveda, root is heating, aphrodisiac, alternative, appetizer, fattening and useful in treatment of piles, biliousness, fatigue, blood related disorders etc. According to Unani system of medicine, root is carminative, tonic, aphrodisiac, antipyretic and useful in bronchitis, ophthalmia, indigestion, vomiting, diarrhea, lumbago, gonorrhea, gleet, hydrophobia, joint pains etc
Indication
1. Itching , Skin disorders , Jaundice , Piles , Decreased sperms count , Frigidity , Dysurea
2. Gonorrhea , General body weakness
Uses
1. Paste – it is used in applying on the skin disorders and where ever there is itching on the skin. . It is used in applying on the pile mass to give instant relief from the pain and burning sensation.
1. Medical smoke – there is a good result of inhaling the smoke of kali musli in respiratory disease like cough, cold and asthma.
1. Powder – its powder is recommended in various ailments like decreased sperm count, impotence general body weakness, loss of stamina and vigor. Besides this it is also used in digestive disorders like indigestion, constipation and loss of appetite. It is very much effective in liver ailments especially in jaundice.
Botanical name: Curculigo orchioides
Family: Liliaceae (Lily family) (Hypoxidaceae)
Common name: Orchid palm grass • Hindi: Kali musli • Oriya: Tala-muli • Kannada: Nela tengu • M General Description: Kali Musli grows as Forest herb. Since generations, it is in use as folk medicine. In many parts of India, due to its over exploitation, Kali Musli is becoming rare in occurrence.
Botanical Description: Kali Musli is a herbaceous tuberous perennial with a short or elongate root stock bearing several fleshy lateral roots; Leaves sessile or petiolate 15-45x1.3-2.5 cm, linear or linear lanceolate, tips sometimes rooting, scape very short, clavate; Flowers in racemes, distichious, yellow, lowest in the racemes 2 sexual, perianth segments elliptic, oblong, hairy on the back; Fruits capsules, derived from inferior tricarpellary syncarpous ovary, 1-4 seeded; Seeds black, oblong, deeply grooved in wavy lines.
Useful Parts: All parts.
Habitat
It is found in places that are up to 6000 feet above sea level. It is more commonly seen in temperate zonal climate. It is seen in countries like Africa, Middle East, Arabia, Pakistan and southern Asian Island. In India it is found every where but predominantly found in Deccan plateau area of Chota Nagpur. It is also commonly seen in south India.
Morphology
It is a perennial shrub having short or elongated fleshy roots. Leaves are 6 to 18 inch in length and ½ to 1 ½ inch broad. It is speared shaped and bears stripes on it. The apex of the leaves is rooting. Flowers present the racemes presentation that is 1 inch in length. Flowers are ½ to 2/3 inch in diameter, shiny and are just above the ground. . Fruit is ½ inch in length, capsulated, ovate in shape, sharp at the apex and contains 1 to 4 seeds in it. Seeds are shiny, oblong in shape, 1/8 inch long, striped and sharp at the apex and base. Rhizome is 1 feet in length and is pulpy. The plant flowers in summers
Chemical Constituents
It contains starch 43.48 %, tannins 4.15 %, enzymes 14.18 % and ash 8.6 %. Besides these it also contains glycoside, orcinol-1-O-beta-D-apiofuranosyl-(1-->6)-beta-D-glucopyranoside, curculigoside, syringic acid, curculigoside and curculigoside C
Medicinal uses:
The rhizomes of the plants are used for the treatment of decline in strength, jaundice and asthma. According to Ayurveda, root is heating, aphrodisiac, alternative, appetizer, fattening and useful in treatment of piles, biliousness, fatigue, blood related disorders etc. According to Unani system of medicine, root is carminative, tonic, aphrodisiac, antipyretic and useful in bronchitis, ophthalmia, indigestion, vomiting, diarrhea, lumbago, gonorrhea, gleet, hydrophobia, joint pains etc
Indication
1. Itching , Skin disorders , Jaundice , Piles , Decreased sperms count , Frigidity , Dysurea
2. Gonorrhea , General body weakness
Uses
1. Paste – it is used in applying on the skin disorders and where ever there is itching on the skin. . It is used in applying on the pile mass to give instant relief from the pain and burning sensation.
1. Medical smoke – there is a good result of inhaling the smoke of kali musli in respiratory disease like cough, cold and asthma.
1. Powder – its powder is recommended in various ailments like decreased sperm count, impotence general body weakness, loss of stamina and vigor. Besides this it is also used in digestive disorders like indigestion, constipation and loss of appetite. It is very much effective in liver ailments especially in jaundice.
Kantakari
Kantakari
Kantakari is a prickly, branches perennial herb with yellow, shining prickles of about 1.5 cm in size. It has very prickly, sparsely hairy, egg shaped leaves; purple flowers, round fruits, yellow in color with green veins and numerous smooth seeds. The branches are densely covered with minute star shaped hairs.
Kantakari is also known as Indian solanum. The fruit of the plant constitutes the drug. The drug is bitter in taste and a mild purgative. Experiments have shown that the fruits and shoots of the plant possess antibacterial properties.
Botanical Name: Solanum xanthocarpum
Family : Solanaceae
Indian Name: Kateli
Chemical constituents
The fruit of the plant yield carpersteral, glucoside-alkaloids and solanocarpine. It also yields glucoside-alkaloids, solamine-S. On hydrolysis it yield alkaloid solanidine-S.
Health Benefits of Kantakari
- Kantakari is beneficial in clearing catarrh and phlegm from the bronchial tubes. It is therefore used in the treatment of respiratory diseases like asthma, bronchitis and cough.
- Kantakari is a valuable remedy in the treatment of dropsy, a disease marked by an excessive collection of fluids in the tissues and cavities or natural hollows of the body. The drug helps increase the secretion and discharge of urine.
- It is also effective in throat disorders like sore throat and tonsillitis. An extract of the plant should be used as a gargle in such cases. This is prepared by continuously boiling the plant in about 2 liters of water after washing it thoroughly, till it reduces to half its volume and it should be filtered.
- It is useful in the treatment of constipation and flatulence. It strengthens the stomach and promotes its action. It corrects disordered processes of nutrition by which the organization ingests, digests, absorbs, utilizes and excretes food substances and restores normal function of the system. The drug also possesses anthelminitic (worm destroying) property and is useful in eliminating intestinal worms.
- The extract of kantakari, prepared as for throat disorders, is also very beneficial in gum diseases. For better results, black mustard should be boiled along with the plant.
- The drug is also effective in treating several other diseases like heart disease, chest pain, certain types of fever, gonorrhea, dysuria, enlargement of the liver, muscular pains and spleen and stone in the urinary bladder. The fruit of the plant is also considered useful in treating sore throat, bronchitis, muscular pains and fevers.
- Kantakari root has been traditionally used in snake and scorpion bites. A paste of the root can be prepared by grinding it on a stone with lemon juice and applying to the affected part. The patient should be taken to the doctor immediately.
Medicinal uses:
It is bitter, pungent, hot, digestant, carminative, diuretic, expectorant and used in cough, asthma, dyspnoea, fever, pleurisy, heart diseases, hoarseness of voice, calculus. Its seeds are analgesic in property..
Uses of Kantakari
Tender leaves and fruits of kantakari are eaten as a v
Kantakari is a prickly, branches perennial herb with yellow, shining prickles of about 1.5 cm in size. It has very prickly, sparsely hairy, egg shaped leaves; purple flowers, round fruits, yellow in color with green veins and numerous smooth seeds. The branches are densely covered with minute star shaped hairs.
Kantakari is also known as Indian solanum. The fruit of the plant constitutes the drug. The drug is bitter in taste and a mild purgative. Experiments have shown that the fruits and shoots of the plant possess antibacterial properties.
Botanical Name: Solanum xanthocarpum
Family : Solanaceae
Indian Name: Kateli
Chemical constituents
The fruit of the plant yield carpersteral, glucoside-alkaloids and solanocarpine. It also yields glucoside-alkaloids, solamine-S. On hydrolysis it yield alkaloid solanidine-S.
Health Benefits of Kantakari
- Kantakari is beneficial in clearing catarrh and phlegm from the bronchial tubes. It is therefore used in the treatment of respiratory diseases like asthma, bronchitis and cough.
- Kantakari is a valuable remedy in the treatment of dropsy, a disease marked by an excessive collection of fluids in the tissues and cavities or natural hollows of the body. The drug helps increase the secretion and discharge of urine.
- It is also effective in throat disorders like sore throat and tonsillitis. An extract of the plant should be used as a gargle in such cases. This is prepared by continuously boiling the plant in about 2 liters of water after washing it thoroughly, till it reduces to half its volume and it should be filtered.
- It is useful in the treatment of constipation and flatulence. It strengthens the stomach and promotes its action. It corrects disordered processes of nutrition by which the organization ingests, digests, absorbs, utilizes and excretes food substances and restores normal function of the system. The drug also possesses anthelminitic (worm destroying) property and is useful in eliminating intestinal worms.
- The extract of kantakari, prepared as for throat disorders, is also very beneficial in gum diseases. For better results, black mustard should be boiled along with the plant.
- The drug is also effective in treating several other diseases like heart disease, chest pain, certain types of fever, gonorrhea, dysuria, enlargement of the liver, muscular pains and spleen and stone in the urinary bladder. The fruit of the plant is also considered useful in treating sore throat, bronchitis, muscular pains and fevers.
- Kantakari root has been traditionally used in snake and scorpion bites. A paste of the root can be prepared by grinding it on a stone with lemon juice and applying to the affected part. The patient should be taken to the doctor immediately.
Medicinal uses:
It is bitter, pungent, hot, digestant, carminative, diuretic, expectorant and used in cough, asthma, dyspnoea, fever, pleurisy, heart diseases, hoarseness of voice, calculus. Its seeds are analgesic in property..
Uses of Kantakari
Tender leaves and fruits of kantakari are eaten as a v
Streptokinase
Streptokinase
Streptokinase is an extracellular metallo-enzyme produced by beta-haemolytic streptococcus and is used as an effective and cheap clot-dissolving medication in some cases of myocardial infarction (heart attack) and pulmonary embolism.
It belongs to a group of medications known as fibrinolytics, and works by activating plasminogen through cleavage to produce plasmin.
The half life of streptokinase is approximately 20 minutes (quoted in SPC).
Mechanism of action
Plasmin is produced in the blood to break down the major constituent of blood clots fibrin, therefore dissolving clots once they have fulfilled their purpose in stopping bleeding. Extra production of plasmin caused by streptokinase breaks down unwanted blood clots, for example, in the lungs (pulmonary embolism). Streptokinase forms a complex in the plasma with plasminogen to form an activator complex. This complex then forms plasmin from unbound plasminogen.[1]
Administration
It is given intravenously as soon as possible after the onset of a heart attack (acute phase - myocardial infarction) to dissolve clots in the arteries of the heart wall. This reduces the amount of damage to the heart muscle. Streptokinase is a bacterial product so the body will build up an immunity to it. It is recommended that this medication should not be used again after four days from the first administration, as it may not be as effective and can also cause an allergic reaction. For this reason, it is usually given only for a person's first heart attack. Further thrombotic events could be treated with tPA. Overdose of Streptokinase or tPA can be treated with Aminocaproic acid.
Streptokinase is an extracellular metallo-enzyme produced by beta-haemolytic streptococcus and is used as an effective and cheap clot-dissolving medication in some cases of myocardial infarction (heart attack) and pulmonary embolism.
It belongs to a group of medications known as fibrinolytics, and works by activating plasminogen through cleavage to produce plasmin.
The half life of streptokinase is approximately 20 minutes (quoted in SPC).
Mechanism of action
Plasmin is produced in the blood to break down the major constituent of blood clots fibrin, therefore dissolving clots once they have fulfilled their purpose in stopping bleeding. Extra production of plasmin caused by streptokinase breaks down unwanted blood clots, for example, in the lungs (pulmonary embolism). Streptokinase forms a complex in the plasma with plasminogen to form an activator complex. This complex then forms plasmin from unbound plasminogen.[1]
Administration
It is given intravenously as soon as possible after the onset of a heart attack (acute phase - myocardial infarction) to dissolve clots in the arteries of the heart wall. This reduces the amount of damage to the heart muscle. Streptokinase is a bacterial product so the body will build up an immunity to it. It is recommended that this medication should not be used again after four days from the first administration, as it may not be as effective and can also cause an allergic reaction. For this reason, it is usually given only for a person's first heart attack. Further thrombotic events could be treated with tPA. Overdose of Streptokinase or tPA can be treated with Aminocaproic acid.
Hybridoma Techniques Antibodies
Introduction to Hybridoma Techniques
Antibodies
highly specific proteins - bind to "antigens" specificity is a very important property of antibodies some uses of antibodies
Antibodies and therapy New Developments in Monoclonal Antibodies Are Hopeful
differentiate between very similar molecular structures detect presence of various molecular structures diagnose and treat certain cancers neutralize biological toxins (e.g., tetanus toxin) hundreds of clinical lab tests purify mixtures of molecular substances (affinity chromotography) identify microorganisms detect infected cells selectively eliminate cells enhance phagocytosis inhibit cellular interactions interfere with virus infections
antibodies are produced by stimulated B-lymphocytes
one B-lymphocyte secretes antibodies of only one specificity
desire large amounts of a single specific antibody
poly-clonal antibodies (produced in vivo)
mixture of many different antibodies of many different specificities many different B-lymphocytes producing antibodies limited time for antibody production (animal will eventually die) low levels of antibodies (animal normally produces only what is needed by that animal)
expensive to maintain animals
dependent on existing genetics of animal
cannot easily select antibodies with different biological properties
mono-clonal antibodies
eliminate mixture of antibodies by isolating B-lymphocytes in tissue culture
grow one isolated B-lymphocyte ---->one " clone" of identical B-cells
problems encountered
isolated B-lymphocyte will often not remain viable in culture don't divide very rapidly very little antibody is produced
need to "add"
viability in tissue culture rapid cell division increased antibody production
tumor cells
many types remain very viable in tissue culture most divide very rapidly but, DON'T produce any antibodies
Combine properties of
B-lymphocyte
+
Tumor Cell
produces antibodies
tissue culture viability rapid cell division
"fuse" B-lymphocyte + tumor cell into one cell
---> “hybrid" cell
culture hybrid cell
continue to grow indefinitely in culture rapid cell division (produces one "clone") antibody production (by one[mono] clone) large amounts of antibodies secreted can manipulate genetics of isolated lymphocytes can select antibodies with certain biological properties much less expensive than maintaining animals
Antibodies
highly specific proteins - bind to "antigens" specificity is a very important property of antibodies some uses of antibodies
Antibodies and therapy New Developments in Monoclonal Antibodies Are Hopeful
differentiate between very similar molecular structures detect presence of various molecular structures diagnose and treat certain cancers neutralize biological toxins (e.g., tetanus toxin) hundreds of clinical lab tests purify mixtures of molecular substances (affinity chromotography) identify microorganisms detect infected cells selectively eliminate cells enhance phagocytosis inhibit cellular interactions interfere with virus infections
antibodies are produced by stimulated B-lymphocytes
one B-lymphocyte secretes antibodies of only one specificity
desire large amounts of a single specific antibody
poly-clonal antibodies (produced in vivo)
mixture of many different antibodies of many different specificities many different B-lymphocytes producing antibodies limited time for antibody production (animal will eventually die) low levels of antibodies (animal normally produces only what is needed by that animal)
expensive to maintain animals
dependent on existing genetics of animal
cannot easily select antibodies with different biological properties
mono-clonal antibodies
eliminate mixture of antibodies by isolating B-lymphocytes in tissue culture
grow one isolated B-lymphocyte ---->one " clone" of identical B-cells
problems encountered
isolated B-lymphocyte will often not remain viable in culture don't divide very rapidly very little antibody is produced
need to "add"
viability in tissue culture rapid cell division increased antibody production
tumor cells
many types remain very viable in tissue culture most divide very rapidly but, DON'T produce any antibodies
Combine properties of
B-lymphocyte
+
Tumor Cell
produces antibodies
tissue culture viability rapid cell division
"fuse" B-lymphocyte + tumor cell into one cell
---> “hybrid" cell
culture hybrid cell
continue to grow indefinitely in culture rapid cell division (produces one "clone") antibody production (by one[mono] clone) large amounts of antibodies secreted can manipulate genetics of isolated lymphocytes can select antibodies with certain biological properties much less expensive than maintaining animals
PRODUCTION OF VACCINES
PRODUCTION OF VACCINES
3.1 Process of Vaccine Production
Materials for vaccines are generally pathogens, bacteria and virus,
use infectious diseases. The quality control of vaccines is most important and regulated by the Minimum Requirements for Biological Products. Each vaccine is controlled by the standards for biological substance production and specific processes and techniques are requested. Vaccines being used in Japan should clear these regulations and pass the tests by the National Institute of health (N.I.H.). Vaccines are classified into live vaccines, inactivated vaccines, and toxoids, as shown in Table 3.1. Recently, recombinant vaccines have been developed using the DNA recombination technique. Processes of the vaccine production are shown in Figures 3.1, 3.2, and 3.3.
3.1.1 Materials for vaccines
a. Vaccine strain: A specified strain of pathogen is nominated as the vaccine strain by the Standard. Influenza virus, however, mutate easily, so the vaccine strain is decided every year by N.I.H. according to the prevalence strain isolated from patients, antigenic analysis, and antibodies found in the patients' blood. The vaccine strain of the year is distributed by N.I.H. Vaccine strains for the live vaccines are kept as the seed lots in N.I.H. The number of transfers of the culture from the seed lot is limited 5 at the maximum.
Media for Vaccine Production
Cultured cells have long been known to serve as excellent hosts for propagation of many types of viruses. The ability of cell culture systems to produce large quantities of attenuated viral particles has served as the basis for the production of both human and veterinary vaccines. Traditional methods have relied on the production of viral agents in cells cultured in medium supplemented with serum, most commonly fetal bovine serum. The animal serum in cell cultures used in production processes can cause a number of problems for manufacturers. The increased costs of raw materials and post-production processing associated with serum have prompted interest in the development of serum-free media for vaccine production. More recently, the potential for contamination by adventitious agents present in serum has heightened regulatory concerns regarding the use of animal-derived components in media used for pharmaceutical manufacturing. With these factors in mind, Sigma has developed new media for vaccine production with reduced levels or completely devoid of animal-derived components. At the same time, we will continue our commitment to traditional manufacturing methods by maintaining and expanding the range of basal media for use with serum supplementation and serve as the basis for customization into serum-free formulations.
The ability to genetically engineer viral particles for use as therapeutic agents to treat genetic diseases (gene therapy) represents one exciting new avenue that science and technology are bringing to modern medicine. Cultured cells have long been known to serve as excellent hosts for the propagation of many types of viruses. Traditional methods have relied on the growth of viral agents in cells cultured in serum-supplemented media. A great concern is the potential for contamination by adventitious agents introduced into the manufacturing process through the use of animal-derived materials. This has heightened regulatory concerns regarding the use of such components in media employed in pharmaceutical manufacturing, particularly in the case of therapeutic injectables. These concerns have led to recommendations that all animal-derived components be avoided when therapeutic agents are manufactured. It is likely that these recommendations will become rigid requirements in the near future. With these factors in mind, Sigma has developed new media tailored to the needs of two of the more popular cell lines used for the propagation of viral particles employed in gene therapy. These media are formulated without the use of animal-derived components.
Vaccine Production in Cells
For decades, vaccines have provided effective protection from influenza for Americans. While they have traditionally been produced in chicken eggs, a new technology-cell-based vaccine production-could save hundreds of thousands of lives in the event of an outbreak of pandemic influenza, or some other infectious disease.
The new approach would use mammalian cells (kidney cells are often used) to grow the influenza viruses. Cell-based vaccine production could more easily meet "surge capacity needs" because cells could be frozen and stored in advance of an epidemic or developed rapidly in response to an epidemic. Cell-based vaccine production dramatically reduces the possibility for contamination and promises to be more reliable, flexible, and expandable than egg-based methods.
In place of eggs, cell-based vaccine production utilizes laboratory-grown cell lines that are capable of hosting a growing virus. The virus is injected into the cells where it multiplies. The cells' outer walls are removed, harvested, purified, and inactivated. A vaccine can be produced in a matter of weeks. Polio vaccine is currently produced using the cell-based method.
While both methods could produce an equally effective vaccine against a virus such as H5N1, egg-based production is physically limited by the availability of specialized eggs and alone may not be able to meet the accelerated demands of a global influenza pandemic. Cell-based vaccines offer the potential to increase production surge capacity and save lives:
• In order to produce 300 million doses of vaccine, egg-based production would require some 900 million eggs. In the case of an avian flu pandemic, egg-producing flocks could decline, jeopardizing vaccine production capabilities.
• While eggs are perishable, cell lines can be safely kept frozen indefinitely, increasing the capability to rapidly produce vaccines if an influenza pandemic were to occur.
• Vaccine manufacturers are able to bypass the steps needed to adapt the virus strains to grow in eggs
• People allergic to eggs cannot receive vaccines produced from chicken eggs, but can be immunized with a cell-based vaccine.
In March 2005, the Department of Health and Human Services issued a five-year contract to Sanofi-Pasteur for $97.1 million to develop cell-based influenza vaccine technology and conduct clinical trials, with the goal of obtaining an FDA license for this vaccine. Under this advanced development contract, the company has committed to develop a plan to establish a U.S. cell-based influenza vaccine manufacturing facility, capable of producing at least 300 million doses of a pandemic influenza vaccine over a one year period.
In May 2006, HHS awarded five contracts totaling more than $1 billion to accelerate development and production of new technologies for influenza vaccines within the U.S. These five contracts support the advanced development of cell-based production technologies for influenza vaccines and will help to modernize and strengthen the nation's influenza vaccine production by creating an alternative to producing influenza vaccines in eggs. The funds are part of $3.3 billion proposed by the President and appropriated by Congress to HHS for fiscal year 2006 to help the nation prepare for a pandemic. (News release).
There is no H5N1 pandemic so it has not been possible to develop an H5N1 pandemic vaccine; however, "pre-pandemic vaccines" have been created, are being refined and tested, and do have some promise both in furthering research and preparedness for a possible pandemic.
3.1 Process of Vaccine Production
Materials for vaccines are generally pathogens, bacteria and virus,
use infectious diseases. The quality control of vaccines is most important and regulated by the Minimum Requirements for Biological Products. Each vaccine is controlled by the standards for biological substance production and specific processes and techniques are requested. Vaccines being used in Japan should clear these regulations and pass the tests by the National Institute of health (N.I.H.). Vaccines are classified into live vaccines, inactivated vaccines, and toxoids, as shown in Table 3.1. Recently, recombinant vaccines have been developed using the DNA recombination technique. Processes of the vaccine production are shown in Figures 3.1, 3.2, and 3.3.
3.1.1 Materials for vaccines
a. Vaccine strain: A specified strain of pathogen is nominated as the vaccine strain by the Standard. Influenza virus, however, mutate easily, so the vaccine strain is decided every year by N.I.H. according to the prevalence strain isolated from patients, antigenic analysis, and antibodies found in the patients' blood. The vaccine strain of the year is distributed by N.I.H. Vaccine strains for the live vaccines are kept as the seed lots in N.I.H. The number of transfers of the culture from the seed lot is limited 5 at the maximum.
Media for Vaccine Production
Cultured cells have long been known to serve as excellent hosts for propagation of many types of viruses. The ability of cell culture systems to produce large quantities of attenuated viral particles has served as the basis for the production of both human and veterinary vaccines. Traditional methods have relied on the production of viral agents in cells cultured in medium supplemented with serum, most commonly fetal bovine serum. The animal serum in cell cultures used in production processes can cause a number of problems for manufacturers. The increased costs of raw materials and post-production processing associated with serum have prompted interest in the development of serum-free media for vaccine production. More recently, the potential for contamination by adventitious agents present in serum has heightened regulatory concerns regarding the use of animal-derived components in media used for pharmaceutical manufacturing. With these factors in mind, Sigma has developed new media for vaccine production with reduced levels or completely devoid of animal-derived components. At the same time, we will continue our commitment to traditional manufacturing methods by maintaining and expanding the range of basal media for use with serum supplementation and serve as the basis for customization into serum-free formulations.
The ability to genetically engineer viral particles for use as therapeutic agents to treat genetic diseases (gene therapy) represents one exciting new avenue that science and technology are bringing to modern medicine. Cultured cells have long been known to serve as excellent hosts for the propagation of many types of viruses. Traditional methods have relied on the growth of viral agents in cells cultured in serum-supplemented media. A great concern is the potential for contamination by adventitious agents introduced into the manufacturing process through the use of animal-derived materials. This has heightened regulatory concerns regarding the use of such components in media employed in pharmaceutical manufacturing, particularly in the case of therapeutic injectables. These concerns have led to recommendations that all animal-derived components be avoided when therapeutic agents are manufactured. It is likely that these recommendations will become rigid requirements in the near future. With these factors in mind, Sigma has developed new media tailored to the needs of two of the more popular cell lines used for the propagation of viral particles employed in gene therapy. These media are formulated without the use of animal-derived components.
Vaccine Production in Cells
For decades, vaccines have provided effective protection from influenza for Americans. While they have traditionally been produced in chicken eggs, a new technology-cell-based vaccine production-could save hundreds of thousands of lives in the event of an outbreak of pandemic influenza, or some other infectious disease.
The new approach would use mammalian cells (kidney cells are often used) to grow the influenza viruses. Cell-based vaccine production could more easily meet "surge capacity needs" because cells could be frozen and stored in advance of an epidemic or developed rapidly in response to an epidemic. Cell-based vaccine production dramatically reduces the possibility for contamination and promises to be more reliable, flexible, and expandable than egg-based methods.
In place of eggs, cell-based vaccine production utilizes laboratory-grown cell lines that are capable of hosting a growing virus. The virus is injected into the cells where it multiplies. The cells' outer walls are removed, harvested, purified, and inactivated. A vaccine can be produced in a matter of weeks. Polio vaccine is currently produced using the cell-based method.
While both methods could produce an equally effective vaccine against a virus such as H5N1, egg-based production is physically limited by the availability of specialized eggs and alone may not be able to meet the accelerated demands of a global influenza pandemic. Cell-based vaccines offer the potential to increase production surge capacity and save lives:
• In order to produce 300 million doses of vaccine, egg-based production would require some 900 million eggs. In the case of an avian flu pandemic, egg-producing flocks could decline, jeopardizing vaccine production capabilities.
• While eggs are perishable, cell lines can be safely kept frozen indefinitely, increasing the capability to rapidly produce vaccines if an influenza pandemic were to occur.
• Vaccine manufacturers are able to bypass the steps needed to adapt the virus strains to grow in eggs
• People allergic to eggs cannot receive vaccines produced from chicken eggs, but can be immunized with a cell-based vaccine.
In March 2005, the Department of Health and Human Services issued a five-year contract to Sanofi-Pasteur for $97.1 million to develop cell-based influenza vaccine technology and conduct clinical trials, with the goal of obtaining an FDA license for this vaccine. Under this advanced development contract, the company has committed to develop a plan to establish a U.S. cell-based influenza vaccine manufacturing facility, capable of producing at least 300 million doses of a pandemic influenza vaccine over a one year period.
In May 2006, HHS awarded five contracts totaling more than $1 billion to accelerate development and production of new technologies for influenza vaccines within the U.S. These five contracts support the advanced development of cell-based production technologies for influenza vaccines and will help to modernize and strengthen the nation's influenza vaccine production by creating an alternative to producing influenza vaccines in eggs. The funds are part of $3.3 billion proposed by the President and appropriated by Congress to HHS for fiscal year 2006 to help the nation prepare for a pandemic. (News release).
There is no H5N1 pandemic so it has not been possible to develop an H5N1 pandemic vaccine; however, "pre-pandemic vaccines" have been created, are being refined and tested, and do have some promise both in furthering research and preparedness for a possible pandemic.
Protease
Protease
A protease is any enzyme that conducts proteolysis, that is, begins protein catabolism by hydrolysis of the peptide bonds that link amino acids together in the polypeptide chain.
Classification
Proteases are currently classified into six groups:
• Serine proteases
• Threonine proteases
• Cysteine proteases
• Aspartic acid proteases
• Metalloproteases
• Glutamic acid proteases
The threonine and glutamic acid proteases were not described until 1995 and 2004, respectively. The mechanism used to cleave a peptide bond involves making an amino acid residue that has the cysteine and threonine (peptidases) or a water molecule (aspartic acid, metallo- and glutamic acid peptidases) nucleophilic so that it can attack the peptide carbonyl group. One way to make a nucleophile is by a catalytic triad, where a histidine residue is used to activate serine, cysteine, or threonine as a nucleophile.
Occurrence
Proteases occur naturally in all organisms. These enzymes are involved in a multitude of physiological reactions from simple digestion of food proteins to highly-regulated cascades (e.g., the blood-clotting cascade, the complement system, apoptosis pathways, and the invertebrate prophenoloxidase-activating cascade). Peptidases can either break specific peptide bonds (limited proteolysis), depending on the amino acid sequence of a protein, or break down a complete peptide to amino acids (unlimited proteolysis). The activity can be a destructive change, abolishing a protein's function or digesting it to its principal components; it can be an activation of a function, or it can be a signal in a signaling pathway.
Bacteria also secrete proteases to hydrolyse the peptide bonds in proteins and therefore break the proteins down into their constituent monomers.
Proteases are also a type of exotoxin, which is a virulence factor in bacteria pathogenesis. Bacteria exotoxic proteases destroy extracellular structures. Protease enzymes are also found used extensively in the bread industry in Bread improver.
Proteases, also known as proteinases or proteolytic enzymes, are a large group of enzymes. Proteases belong to the class of enzymes known as hydrolases, which catalyse the reaction of hydrolysis of various bonds with the participation of a water molecule.
Proteases are involved in digesting long protein chains into short fragments, splitting the peptide bonds that link amino acid residues. Some of them can detach the terminal amino acids from the protein chain (exopeptidases, such as aminopeptidases, carboxypeptidase www A); the others attack internal peptide bonds of a protein (endopeptidases, such as trypsin, chymotrypsin, pepsin, papain, elastase).
Proteases are divided into four major groups according to the character of their catalytic active site and conditions of action: serine proteinases, cysteine (thiol) proteinases, aspartic proteinases, and metalloproteinases. Attachment of a protease to a certain group depends on the structure of catalytic site and the amino acid (as one of the constituents) essential for its activity.
Proteases are used throughout an organism for various metabolic processes. Acid proteases secreted into the stomach (such as pepsin) and serine proteases present in duodenum (trypsin and chymotrypsin) enable us to digest the protein in food; proteases present in blood serum (thrombin, plasmin, Hageman factor, etc.) play important role in blood-clotting, as well as lysis of the clots, and the correct action of the immune system. Other proteases are present in leukocytes (elastase, cathepsin G) and play several different roles in metabolic control. Proteases determine the lifetime of other proteins playing important physiological role like hormones, antibodies, or other enzymes -- this is one of the fastest "switching on" and "switching off" regulatory mechanisms in the physiology of an organism. By complex cooperative action the proteases may proceed as cascade reactions, which result in rapid and efficient amplification of an organism's response to a physiological signal.
Inhibitors
The function of peptidases is inhibited by protease inhibitor enzymes. Examples of protease inhibitors are the class of serpins (serine protease or peptidase inhibitors), incorporating alpha 1-antitrypsin. Other serpins are complement 1-inhibitor, antithrombin, alpha 1-antichymotrypsin, plasminogen activator inhibitor 1 (coagulation, fibrinolysis) and the recently discovered neuroserpin.
Natural protease inhibitors include the family of lipocalin proteins, which play a role in cell regulation and differentiation. Lipophilic ligands, attached to lipocalin proteins, have been found to possess tumor protease inhibiting properties. The natural protease inhibitors are not to be confused with the protease inhibitors used in antiretroviral therapy. Some viruses, with HIV among them, depend on proteases in their reproductive cycle. Thus, protease inhibitors are developed as antiviral means.
Degradation
Proteases, being themselves proteins, are known to be cleaved by other protease molecules, sometimes of the same variety. This may be an important method of regulation of peptidase activity.
Serine protease
Crystal structure of Trypsin, a typical serine protease.
Serine proteases or serine endopeptidases (newer name) are proteases (enzymes that cut peptide bonds in proteins) in which one of the amino acids at the active site is serine.
They are found in both single-cell and complex organisms, in both cells with nuclei (eukaryotes) and without nuclei (prokaryotes).
Serine proteases are grouped into clans that share structural similarities (homology) and are then further subgrouped into families with simiar sequences.
The major clans found in humans include the chymotrypsin-like, the subtilisin-like, the alpha/beta hydrolase, and signal peptidase clans.
In evolutionary history, serine proteases were originally digestive enzymes. In mammals, they evolved by gene duplication to serve functions in blood clotting, the immune system, and inflammation.
Serine proteases are paired with serine protease inhibitors, which turn off their activity when they are no longer needed.[1]
Digestive serine proteases
Members
Chymotrypsin-clan
The three serine proteases of the chymotrypsin-like clan that have been studied in greatest detail are chymotrypsin, trypsin, and elastase. All three enzymes are synthesized by the pancreatic acinar cells, secreted in the small intestine, and are responsible for catalyzing the hydrolysis of peptide bonds. All three of these enzymes are similar in structure, as shown through their X-ray structures. The differing aspect lies in the peptide bond that is being cleaved; this is called the scissile bond. The different enzymes, like most enzymes, are highly specific in the reactions they catalyze. Each of these digestive serine proteases targets different regions of a polypeptide chain, based upon the side chains of the amino acid residues surrounding the site of cleavage:
• Chymotrypsin is responsible for cleaving peptide bonds following a bulky hydrophobic amino acid residue. Preferred residues include phenylalanine, tryptophan, and tyrosine, which fit into a snug hydrophobic pocket.
• Trypsin is responsible for cleaving peptide bonds following a positively-charged amino acid residue. Instead of having the hydrophobic pocket of the chymotrypsin, there exists an aspartic acid residue at the base of the pocket. This can then interact with positively-charged residues such as arginine and lysine on the substrate peptide to be cleaved.
• Elastase is responsible for cleaving peptide bonds following a small neutral amino acid residue, such as Alanine, glycine, and valine. (These amino acid residues form much of the connective tissues in meat). The pocket that is in "trypsin" and "chymotrypsin" is now partially filled with valine and threonine, rendering it a mere depression, which can accommodate these smaller amino acid residues.
The combination of these three enzymes make an incredibly effective digestive team and are primarily responsible for the digestion of proteins.
Subtilisin
Subtilisin is a serine protease in prokaryotes. Subtilisin is evolutionary unrelated to the chymotrypsin-clan, but shares the same catalytic mechanism utilising a catalytic triad, to create a nucleophilic serine. This is the classic example used to illustrate convergent evolution, since the same mechanism evolved twice independently during evolution.
Catalytic mechanism
The main player in the catalytic mechanism in the chymotrypsin and subtillisin clan enzymes mentioned above is the catalytic triad. The triad is located in the active site of the enzyme, where catalysis occurs, and is preserved in all serine protease enzymes. The triad is a coordinated structure consisting of three essential amino acids: histidine (His 57), serine (Ser 195) (hence the name "serine protease") and aspartic acid (Asp 102). Located very near one another near the heart of the enzyme, these three key amino acids each play an essential role in the cleaving ability of the proteases.
In the event of catalysis, an ordered mechanism occurs in which several intermediates are generated. The catalysis of the peptide cleavage can be seen as a ping-pong catalysis, in which a substrate binds (in this case, the polypeptide being cleaved), a product is released (the N-terminus "half" of the peptide), another substrate binds (in this case, water), and another product is released (the C-terminus "half" of the peptide).
Each amino acid in the triad performs a specific task in this process:
*The serine has an -OH group that is able to act as a nucleophile, attacking the carbonyl carbon of the scissile peptide bond of the substrate.
• A pair of electrons on the histidine nitrogen has the ability to accept the hydrogen from the serine -OH group, thus coordinating the attack of the peptide bond.
• The carboxyl group on the aspartic acid in turn hydrogen bonds with the histidine, making the pair of electrons mentioned above much more electronegative.
The whole reaction can be summarized as follows:
• The polypeptide substrate binds to the surface of the serine protease enzyme such that scissile bond is inserted into the active site of the enzyme, with the carbonyl carbon of this bond positioned near the nucleophilic serine.
• The serine -OH attacks the carbonyl carbon, and the nitrogen of the histidine accepts the hydrogen from the -OH of the [serine] and a pair of electrons from the double bond of the carbonyl oxygen moves to the oxygen. As a result, a tetrahedral intermediate is generated.
• The bond joining the nitrogen and the carbon in the peptide bond is now broken. The covalent electrons creating this bond move to attack the hydrogen of the histidine, breaking the connection. The electrons that previously moved from the carbonyl oxygen double bond move back from the negative oxygen to recreate the bond, generating an acyl-enzyme intermediate.
• Now, water comes in to the reaction. Water replaces the N-terminus of the cleaved peptide, and attacks the carbonyl carbon. Once again, the electrons from the double bond move to the oxygen making it negative, as the bond between the oxygen of the water and the carbon is formed. This is coordinated by the nitrogen of the histidine. which accepts a proton from the water. Overall, this generates another tetrahedral intermediate.
• In a final reaction, the bond formed in the first step between the serine and the carbonyl carbon moves to attack the hydrogen that the histidine just acquired. The now electron-deficient carbonyl carbon re-forms the double bond with the oxygen. As a result, the C-terminus of the peptide is now ejected.
Additional stabilizing effects
It was discovered that additional amino acids of the protease, Gly 193 and Ser 195, are involved in creating what is called an oxyanion hole. Both Gly 193 and Ser 195 can donate backbone hydrogens for hydrogen bonding. When the tetrahedral intermediate of step 1 and step 3 are generated, the negative oxygen ion, having accepted the electrons from the carbonyl double bond fits perfectly into the oxyanion hole. In effect, serine proteases preferentially bind the transition state and the overall structure is favored, lowering the activation energy of the reaction. This "preferential binding" is responsible for much of the catalytic efficiency of the enzyme.
Zymogens
There are certain inhibitors which resemble the tetrahedral intermediate, and thus fill up the active site, preventing the enzyme from working properly. Trypsin, a powerful digestive enzyme, is generated in the pancreas. Inhibitors prevent self-digestion of the pancreas itself.
Zymogens are the usually inactive precursors of an enzyme. If the digestive enzymes were active when synthesized, they would immediately start chewing up the synthesizing organs and tissues. Acute pancreatitis is such a condition, in which there is premature activation of the digestive enzymes in the pancreas, resulting in self-digestion (autolysis). It also complicates postmortem investigations, as the pancreas often digests itself before it can be assessed visually.
Zymogens are large, inactive structures, which have the ability to break apart or change into the smaller activated enzymes. The difference between zymogens and the activated enzymes lies in the fact that the active site for catalysis of the zymogens is distorted. As a result, the substrate polypeptide cannot bind effectively, and proteolysis does not occur. Only after activation, during which the conformation and structure of the zymogen change and the active site is opened, can proteolysis occur.
Zymogen Enzyme Notes
Trypsinogen trypsin
When trypsinogen enters the small intestine from the pancreas, secretions from the duodenal mucosa cleaves the lysine 15 - isoleucine 16 peptide bond of the zymogen. As a result, the zymogen trypsinogen breaks down into trypsin. Recall that trypsin is also responsible for cleaving lysine peptide bonds, and thus, once a small amount of trypsin is generated, it participates in cleavage of its own zymogen, generating even more trypsin. The process of trypsin activation can thus be called autocatalytic.
Chymotrypsinogen chymotrypsin
After the Arg 15 - Ile 16 bond in the chymotrypsinogen zymogen is cleaved by trypsin, the newly generated structure called a pi-chymotrypsin undergoes autolysis (self digestion), yielding active chymotrypsin.
Proelastase elastase
It is activated by cleavage through trypsin.
As can be seen, trypsinogen activation to trypsin is essential, because it activates its own reaction, as well as the reaction of both chymotrypsin and elastase. It is therefore essential that this activation doesn't occur prematurely. There are several protective measures taken by the organism to prevent self-digestion:
• The activation of trypsinogen by trypsin is relatively slow
• The zymogens are stored in zymogen granules, capsules that have walls that are thought to be resistant to proteolysis.
Inhibition
Serine proteases are inhibited by a diverse group of inhibitors, including synthetic chemical inhibitors for research or therapeutic purposes, and also natural proteinaceous inhibitors. One family of natural inhibitors called "serpins" (abbreviated from serine protease inhibitors) can form a covalent bond with the serine protease, inhibiting its function. The best-studied serpins are antithrombin and alpha 1-antitrypsin, studied for their role in coagulation/thrombosis and emphysema/A1AT respectively. Artificial irreversible small molecule inhibitors include AEBSF and PMSF.
Role in disease
Mutations may lead to decreased or increased activity of enzymes. This may have different consequences, depending on the normal function of the serine protease. For example, mutations in protein C, when leading to insufficient protein levels or activity, predispose to thrombosis.
Diagnostic use
Determination of serine protease levels may be useful in the context of particular diseases.
• Coagulation factor levels may be required in the diagnosis of hemorrhagic or thrombotic conditions.
• Fecal elastase is employed to determine the exocrine activity of the pancreas, e.g. in cystic fibrosis or chronic pancreatitis.
• Prostate specific antigen is used to determine prostate cancer risk
Aspartate protease
Aspartic proteases are a family of eukaryotic protease enzymes that utilize an aspartate residue for catalysis of their peptide substrates. In general, they have two highly-conserved aspartates in the active site and are optimally active at acidic pH. Nearly all known aspartyl proteases are inhibited by pepstatin.
Eukaryotic aspartic proteases include pepsins, cathepsins, and renins. They have a two-domain structure, probably arising from ancestral duplication. Retroviral and retrotransposon proteases (Pfam PF00077) are much smaller and appear to be homologous to a single domain of the eukaryotic aspartyl proteases.
Examples
• HIV-1 protease - a major drug-target for treatment of HIV
• Chymosin (or "rennin", with two "n"s)
• Renin (with one "n")
• Cathepsin D
• Pepsin
• Plasmepsin
Mechanism
Proposed mechanism of peptide cleavage by aspartyl proteases[1]
While a number of different mechanisms for aspartyl proteases have been proposed, the most widely accepted is a general acid-base mechanism involving coordination of a water molecule between the two highly-conserved aspartate residues.[1][2] One aspartate activates the water by abstracting a proton, enabling the water to attack the carbonyl carbon of the substrate scissile bond, generating a tetrahedral oxyanion intermediate. Rearrangement of this intermediate leads to protonation of the scissile amide.
A protease is any enzyme that conducts proteolysis, that is, begins protein catabolism by hydrolysis of the peptide bonds that link amino acids together in the polypeptide chain.
Classification
Proteases are currently classified into six groups:
• Serine proteases
• Threonine proteases
• Cysteine proteases
• Aspartic acid proteases
• Metalloproteases
• Glutamic acid proteases
The threonine and glutamic acid proteases were not described until 1995 and 2004, respectively. The mechanism used to cleave a peptide bond involves making an amino acid residue that has the cysteine and threonine (peptidases) or a water molecule (aspartic acid, metallo- and glutamic acid peptidases) nucleophilic so that it can attack the peptide carbonyl group. One way to make a nucleophile is by a catalytic triad, where a histidine residue is used to activate serine, cysteine, or threonine as a nucleophile.
Occurrence
Proteases occur naturally in all organisms. These enzymes are involved in a multitude of physiological reactions from simple digestion of food proteins to highly-regulated cascades (e.g., the blood-clotting cascade, the complement system, apoptosis pathways, and the invertebrate prophenoloxidase-activating cascade). Peptidases can either break specific peptide bonds (limited proteolysis), depending on the amino acid sequence of a protein, or break down a complete peptide to amino acids (unlimited proteolysis). The activity can be a destructive change, abolishing a protein's function or digesting it to its principal components; it can be an activation of a function, or it can be a signal in a signaling pathway.
Bacteria also secrete proteases to hydrolyse the peptide bonds in proteins and therefore break the proteins down into their constituent monomers.
Proteases are also a type of exotoxin, which is a virulence factor in bacteria pathogenesis. Bacteria exotoxic proteases destroy extracellular structures. Protease enzymes are also found used extensively in the bread industry in Bread improver.
Proteases, also known as proteinases or proteolytic enzymes, are a large group of enzymes. Proteases belong to the class of enzymes known as hydrolases, which catalyse the reaction of hydrolysis of various bonds with the participation of a water molecule.
Proteases are involved in digesting long protein chains into short fragments, splitting the peptide bonds that link amino acid residues. Some of them can detach the terminal amino acids from the protein chain (exopeptidases, such as aminopeptidases, carboxypeptidase www A); the others attack internal peptide bonds of a protein (endopeptidases, such as trypsin, chymotrypsin, pepsin, papain, elastase).
Proteases are divided into four major groups according to the character of their catalytic active site and conditions of action: serine proteinases, cysteine (thiol) proteinases, aspartic proteinases, and metalloproteinases. Attachment of a protease to a certain group depends on the structure of catalytic site and the amino acid (as one of the constituents) essential for its activity.
Proteases are used throughout an organism for various metabolic processes. Acid proteases secreted into the stomach (such as pepsin) and serine proteases present in duodenum (trypsin and chymotrypsin) enable us to digest the protein in food; proteases present in blood serum (thrombin, plasmin, Hageman factor, etc.) play important role in blood-clotting, as well as lysis of the clots, and the correct action of the immune system. Other proteases are present in leukocytes (elastase, cathepsin G) and play several different roles in metabolic control. Proteases determine the lifetime of other proteins playing important physiological role like hormones, antibodies, or other enzymes -- this is one of the fastest "switching on" and "switching off" regulatory mechanisms in the physiology of an organism. By complex cooperative action the proteases may proceed as cascade reactions, which result in rapid and efficient amplification of an organism's response to a physiological signal.
Inhibitors
The function of peptidases is inhibited by protease inhibitor enzymes. Examples of protease inhibitors are the class of serpins (serine protease or peptidase inhibitors), incorporating alpha 1-antitrypsin. Other serpins are complement 1-inhibitor, antithrombin, alpha 1-antichymotrypsin, plasminogen activator inhibitor 1 (coagulation, fibrinolysis) and the recently discovered neuroserpin.
Natural protease inhibitors include the family of lipocalin proteins, which play a role in cell regulation and differentiation. Lipophilic ligands, attached to lipocalin proteins, have been found to possess tumor protease inhibiting properties. The natural protease inhibitors are not to be confused with the protease inhibitors used in antiretroviral therapy. Some viruses, with HIV among them, depend on proteases in their reproductive cycle. Thus, protease inhibitors are developed as antiviral means.
Degradation
Proteases, being themselves proteins, are known to be cleaved by other protease molecules, sometimes of the same variety. This may be an important method of regulation of peptidase activity.
Serine protease
Crystal structure of Trypsin, a typical serine protease.
Serine proteases or serine endopeptidases (newer name) are proteases (enzymes that cut peptide bonds in proteins) in which one of the amino acids at the active site is serine.
They are found in both single-cell and complex organisms, in both cells with nuclei (eukaryotes) and without nuclei (prokaryotes).
Serine proteases are grouped into clans that share structural similarities (homology) and are then further subgrouped into families with simiar sequences.
The major clans found in humans include the chymotrypsin-like, the subtilisin-like, the alpha/beta hydrolase, and signal peptidase clans.
In evolutionary history, serine proteases were originally digestive enzymes. In mammals, they evolved by gene duplication to serve functions in blood clotting, the immune system, and inflammation.
Serine proteases are paired with serine protease inhibitors, which turn off their activity when they are no longer needed.[1]
Digestive serine proteases
Members
Chymotrypsin-clan
The three serine proteases of the chymotrypsin-like clan that have been studied in greatest detail are chymotrypsin, trypsin, and elastase. All three enzymes are synthesized by the pancreatic acinar cells, secreted in the small intestine, and are responsible for catalyzing the hydrolysis of peptide bonds. All three of these enzymes are similar in structure, as shown through their X-ray structures. The differing aspect lies in the peptide bond that is being cleaved; this is called the scissile bond. The different enzymes, like most enzymes, are highly specific in the reactions they catalyze. Each of these digestive serine proteases targets different regions of a polypeptide chain, based upon the side chains of the amino acid residues surrounding the site of cleavage:
• Chymotrypsin is responsible for cleaving peptide bonds following a bulky hydrophobic amino acid residue. Preferred residues include phenylalanine, tryptophan, and tyrosine, which fit into a snug hydrophobic pocket.
• Trypsin is responsible for cleaving peptide bonds following a positively-charged amino acid residue. Instead of having the hydrophobic pocket of the chymotrypsin, there exists an aspartic acid residue at the base of the pocket. This can then interact with positively-charged residues such as arginine and lysine on the substrate peptide to be cleaved.
• Elastase is responsible for cleaving peptide bonds following a small neutral amino acid residue, such as Alanine, glycine, and valine. (These amino acid residues form much of the connective tissues in meat). The pocket that is in "trypsin" and "chymotrypsin" is now partially filled with valine and threonine, rendering it a mere depression, which can accommodate these smaller amino acid residues.
The combination of these three enzymes make an incredibly effective digestive team and are primarily responsible for the digestion of proteins.
Subtilisin
Subtilisin is a serine protease in prokaryotes. Subtilisin is evolutionary unrelated to the chymotrypsin-clan, but shares the same catalytic mechanism utilising a catalytic triad, to create a nucleophilic serine. This is the classic example used to illustrate convergent evolution, since the same mechanism evolved twice independently during evolution.
Catalytic mechanism
The main player in the catalytic mechanism in the chymotrypsin and subtillisin clan enzymes mentioned above is the catalytic triad. The triad is located in the active site of the enzyme, where catalysis occurs, and is preserved in all serine protease enzymes. The triad is a coordinated structure consisting of three essential amino acids: histidine (His 57), serine (Ser 195) (hence the name "serine protease") and aspartic acid (Asp 102). Located very near one another near the heart of the enzyme, these three key amino acids each play an essential role in the cleaving ability of the proteases.
In the event of catalysis, an ordered mechanism occurs in which several intermediates are generated. The catalysis of the peptide cleavage can be seen as a ping-pong catalysis, in which a substrate binds (in this case, the polypeptide being cleaved), a product is released (the N-terminus "half" of the peptide), another substrate binds (in this case, water), and another product is released (the C-terminus "half" of the peptide).
Each amino acid in the triad performs a specific task in this process:
*The serine has an -OH group that is able to act as a nucleophile, attacking the carbonyl carbon of the scissile peptide bond of the substrate.
• A pair of electrons on the histidine nitrogen has the ability to accept the hydrogen from the serine -OH group, thus coordinating the attack of the peptide bond.
• The carboxyl group on the aspartic acid in turn hydrogen bonds with the histidine, making the pair of electrons mentioned above much more electronegative.
The whole reaction can be summarized as follows:
• The polypeptide substrate binds to the surface of the serine protease enzyme such that scissile bond is inserted into the active site of the enzyme, with the carbonyl carbon of this bond positioned near the nucleophilic serine.
• The serine -OH attacks the carbonyl carbon, and the nitrogen of the histidine accepts the hydrogen from the -OH of the [serine] and a pair of electrons from the double bond of the carbonyl oxygen moves to the oxygen. As a result, a tetrahedral intermediate is generated.
• The bond joining the nitrogen and the carbon in the peptide bond is now broken. The covalent electrons creating this bond move to attack the hydrogen of the histidine, breaking the connection. The electrons that previously moved from the carbonyl oxygen double bond move back from the negative oxygen to recreate the bond, generating an acyl-enzyme intermediate.
• Now, water comes in to the reaction. Water replaces the N-terminus of the cleaved peptide, and attacks the carbonyl carbon. Once again, the electrons from the double bond move to the oxygen making it negative, as the bond between the oxygen of the water and the carbon is formed. This is coordinated by the nitrogen of the histidine. which accepts a proton from the water. Overall, this generates another tetrahedral intermediate.
• In a final reaction, the bond formed in the first step between the serine and the carbonyl carbon moves to attack the hydrogen that the histidine just acquired. The now electron-deficient carbonyl carbon re-forms the double bond with the oxygen. As a result, the C-terminus of the peptide is now ejected.
Additional stabilizing effects
It was discovered that additional amino acids of the protease, Gly 193 and Ser 195, are involved in creating what is called an oxyanion hole. Both Gly 193 and Ser 195 can donate backbone hydrogens for hydrogen bonding. When the tetrahedral intermediate of step 1 and step 3 are generated, the negative oxygen ion, having accepted the electrons from the carbonyl double bond fits perfectly into the oxyanion hole. In effect, serine proteases preferentially bind the transition state and the overall structure is favored, lowering the activation energy of the reaction. This "preferential binding" is responsible for much of the catalytic efficiency of the enzyme.
Zymogens
There are certain inhibitors which resemble the tetrahedral intermediate, and thus fill up the active site, preventing the enzyme from working properly. Trypsin, a powerful digestive enzyme, is generated in the pancreas. Inhibitors prevent self-digestion of the pancreas itself.
Zymogens are the usually inactive precursors of an enzyme. If the digestive enzymes were active when synthesized, they would immediately start chewing up the synthesizing organs and tissues. Acute pancreatitis is such a condition, in which there is premature activation of the digestive enzymes in the pancreas, resulting in self-digestion (autolysis). It also complicates postmortem investigations, as the pancreas often digests itself before it can be assessed visually.
Zymogens are large, inactive structures, which have the ability to break apart or change into the smaller activated enzymes. The difference between zymogens and the activated enzymes lies in the fact that the active site for catalysis of the zymogens is distorted. As a result, the substrate polypeptide cannot bind effectively, and proteolysis does not occur. Only after activation, during which the conformation and structure of the zymogen change and the active site is opened, can proteolysis occur.
Zymogen Enzyme Notes
Trypsinogen trypsin
When trypsinogen enters the small intestine from the pancreas, secretions from the duodenal mucosa cleaves the lysine 15 - isoleucine 16 peptide bond of the zymogen. As a result, the zymogen trypsinogen breaks down into trypsin. Recall that trypsin is also responsible for cleaving lysine peptide bonds, and thus, once a small amount of trypsin is generated, it participates in cleavage of its own zymogen, generating even more trypsin. The process of trypsin activation can thus be called autocatalytic.
Chymotrypsinogen chymotrypsin
After the Arg 15 - Ile 16 bond in the chymotrypsinogen zymogen is cleaved by trypsin, the newly generated structure called a pi-chymotrypsin undergoes autolysis (self digestion), yielding active chymotrypsin.
Proelastase elastase
It is activated by cleavage through trypsin.
As can be seen, trypsinogen activation to trypsin is essential, because it activates its own reaction, as well as the reaction of both chymotrypsin and elastase. It is therefore essential that this activation doesn't occur prematurely. There are several protective measures taken by the organism to prevent self-digestion:
• The activation of trypsinogen by trypsin is relatively slow
• The zymogens are stored in zymogen granules, capsules that have walls that are thought to be resistant to proteolysis.
Inhibition
Serine proteases are inhibited by a diverse group of inhibitors, including synthetic chemical inhibitors for research or therapeutic purposes, and also natural proteinaceous inhibitors. One family of natural inhibitors called "serpins" (abbreviated from serine protease inhibitors) can form a covalent bond with the serine protease, inhibiting its function. The best-studied serpins are antithrombin and alpha 1-antitrypsin, studied for their role in coagulation/thrombosis and emphysema/A1AT respectively. Artificial irreversible small molecule inhibitors include AEBSF and PMSF.
Role in disease
Mutations may lead to decreased or increased activity of enzymes. This may have different consequences, depending on the normal function of the serine protease. For example, mutations in protein C, when leading to insufficient protein levels or activity, predispose to thrombosis.
Diagnostic use
Determination of serine protease levels may be useful in the context of particular diseases.
• Coagulation factor levels may be required in the diagnosis of hemorrhagic or thrombotic conditions.
• Fecal elastase is employed to determine the exocrine activity of the pancreas, e.g. in cystic fibrosis or chronic pancreatitis.
• Prostate specific antigen is used to determine prostate cancer risk
Aspartate protease
Aspartic proteases are a family of eukaryotic protease enzymes that utilize an aspartate residue for catalysis of their peptide substrates. In general, they have two highly-conserved aspartates in the active site and are optimally active at acidic pH. Nearly all known aspartyl proteases are inhibited by pepstatin.
Eukaryotic aspartic proteases include pepsins, cathepsins, and renins. They have a two-domain structure, probably arising from ancestral duplication. Retroviral and retrotransposon proteases (Pfam PF00077) are much smaller and appear to be homologous to a single domain of the eukaryotic aspartyl proteases.
Examples
• HIV-1 protease - a major drug-target for treatment of HIV
• Chymosin (or "rennin", with two "n"s)
• Renin (with one "n")
• Cathepsin D
• Pepsin
• Plasmepsin
Mechanism
Proposed mechanism of peptide cleavage by aspartyl proteases[1]
While a number of different mechanisms for aspartyl proteases have been proposed, the most widely accepted is a general acid-base mechanism involving coordination of a water molecule between the two highly-conserved aspartate residues.[1][2] One aspartate activates the water by abstracting a proton, enabling the water to attack the carbonyl carbon of the substrate scissile bond, generating a tetrahedral oxyanion intermediate. Rearrangement of this intermediate leads to protonation of the scissile amide.
General Principles of Toxicology
General Principles of Toxicology
Know the following definitions:
Toxicology: relation of hazardous effects of chemicals, including drugs, to biological systems
Acute toxicity: adverse effect resulting from a single, usually large, exposure to a toxin
Subacute toxicity: Somewhat acute, between acute and chronic (online med. dictionary)
Chronic toxicity: harmful effect from repeated exposures to a toxin for 3+ months
Therapeutic index: LD50/ED50
Risk: probability that injury will result from exposure to a substance for given conditions, dose, and route
Threshold limit values (TLV): maximum safe ambient air concentrations of chemicals during a workweek,a 15 minute interval, and instantaneously
Know the following general mechanisms by which drugs or chemicals can cause toxicity:
Alterations in receptor-ligand interactions
Nicotine, most drugs
Alterations in membrane function
Local anesthetics, hydrocarbons
Interference with cellular energy mechanisms
Cyanide, Pentachlorophenol
Covalent binding to biomolecules
Organophosphates, alkylating agents
Interference with calcium homeostasis
Oxalates
By causing non-lethal alterations in somatic cells
Carcinogens, e.g. aflatoxin
Alterations in ligand-activated transcription factors
Dioxins
By inducing programmed cell death (apoptosis)
Acetaminophen
· Understand the primary determinants of toxicity
Dose and dose rate
Duration of exposure: long duration: bad.
(Principle) routes of exposure: inhalation, transdermal, and oral
· Understand how the following factors modify toxicity
Biotransformation
Parathion à Paraxon
Methanol à Formaldehyde à Formic Acid (MeOH shares pathway with EtOH)
Immune function
Review the hypersensitivities, e.g. Mosby 1-9 or perhaps even your long term memory
Phototoxicity: drug intermediates accumulate in skin, exposed to UV à toxic compounds
Examples: tetracycline, sulfonamides
Age: pharmodynamics, pharmokinetics vary over a lifetime
Gender: no specifics were discussed
· Understand how to manage a poisoned patient - know the ABCD's
Airway: should be cleared of vomitus or other obstruction and an airway or ET tube inserted
Breathing:Assesed by observation and measurements of arterial gases
Intubate and mechanically ventilate if necessary
Circulation
Monitor pulse rate, blood pressure, and urinary output
Start IV and draw blood for glucose and other labs
Dextrose: to every patient with altered mental status (thinking about hypoglycemic problems)
100mg thiamine to alcoholic and malnourished to prevent Wernicke-Korsakoff’s
· Understand the importance of the history and physical examination in treating a poisoned patient
Oral history may be unreliable for a number of reasons
Be on the lookout for classical OD effects of common meds and poisons
Treat the patient, not the poison
· Understand the importance of the following laboratory analysis and procedures
Arterial blood gases
CO2: increased with hypoventilation
PO2: low with aspiration pneumonia or drug-induced pulmonary edema
Reduced with poor tissue oxygenation due to hypoxia, hypotension, or Cn poisoning
May appear normal in CO because dissolved O2 is measured, not oxyhemoglobin
Electrolyte analysis (an ion gap)
(Na+ + K+) + (HCO3
- + Cl-) (this is just major cations minus anions)
normal = 12 plus/minus 4
elevated by:
renal failure
diabetic ketoacidosis
shock-induced lactic adicosis
Drug-induced metabolic acidosis (salicylates, methanol, ethylene glycol, isoniazid, Fe)
Renal and liver function tests
Renal tests: In the UA look for:
BUN, creatinine (nitrogen load and glomerular filtration checks, respectively)
CK and myoglobin (muscle insults)
oxlate crystals: suggest ethylene glycol poisoning
Liver tests: transaminases, look at the PT
Osmolar gap: useful in alcohols poisonings; alcohols will increase the osmolar gap (K. table 59-4)
Calculated serum osmolality: [ 2Na+ + glucose/18 + BUN/3]
Osmolar gap: measured osmolality – calculated osmolality
EKG examples:
Wide QRS: tricyclics, quinidine
Long QT: quinidine, phenothiazine, tricyclics
Variable AV block, screwy rhythms: dig. overdose
Ischemic changes: hypoxemia due to CO
· Understand how toxins can be removed or elimination can be enhanced
Gastric lavage
> 30 minutes have passed since the ingestion of a corrosive material
ingestion of hydrocarbons
coma, stupor, delerium, unconsciousness, convulsions
Induced emesis (know contraindications)
Syrup of Ipecac is often used; same contraindications as above
Increased rate of excretion
Catharsis: increased clearance of intestinal contents
Sorbitol is the preferred agent (hellooooo, apple juice), MgSO4 can be used if kidneys are ok
osmotic diuretics: e.g. mannitol, urea, etc.
Altered urinary pH
Alkalination: useful for salicylate or phenobarbitol overdose
Acidification: not recommended b/c worsens renal effects of rhabdomyolysis
Peritoneal dialysis: simple and available, but inefficient for most drugs
Hemodialysis: especially useful in cases where electrolyte and fluid imbalances are present
Hemoperfusion: blood pumped from pt’s vein through a cartridge filled with adsorbent material
Especially effective for high molecular weight compounds
· Understand the importance of the following specific drugs in managing the poisoned patient
Activated charcoal: large surface area, suggested dose is 10:1 charcoal to est. weight of toxin
Good for: adsorption of many drugs and poisons
No good for: Fe, Li, K, Cn, alcohols, corrosive acids and alkali, methylcarbmate, tolbutamide
Ipecac syrup: emetic agent; use 30 ml for adults, 10-15 ml for kids, repeated q 15 minutes if necessary
Emetic contraindications still apply: not for corrosives, hydrocarbons, rapidly acting convulsants
Ammonium chloride: used to acidify the urine
Not used much clinically because of side effects, e.g. indirect kidney damage
urinary excretion of weak organic bases
Sodium bicarbonate: urine alkalinization
urinary excretion of weak organic acids
Magnesium sulfate: cathartic, contraindicated in renal compromise
Mannitol: used to renal clearance of toxins (also to intraocular and intracranial pressure)
Deferoxamine: use IV or IM; chelator of choice for Fe poisoning
Dimercaprol: single-agent therapy for arsenic and mercury, use for lead with EDTA
Edetate, calcium disodium (EDTA): efficient chelator of many di/trivalent metal ions, esp. lead
Penicillamine: chelator of Cu and Pb
used in Wilson’s cystinuria, resistant cases of rheumatoid arthritis
Succimer (Dimercaptosuccinic Acid, DMSA), analog of dimercaprol,
prevents and reverses metal-induced inhibition of sulfhydryl-containing enzymes
urinary Pb excretion, protects against lethal effects of As, kidney [Hg]
Acetylcysteine: acetaminophen OD antidote, give within 8-10 hours of insult
Digoxin-specific FAb antibody: binds Dig
Atropine: used for cholinesterase poisonings to block ACh binding
Pralidoxime (2-PAM): cholinesterase reactivator, given only for organophosphates
Physostigmine: suggested for antimuscarinic, anticholinergic agents, but not for TCA’s
Flumazenil: used for benzodiazepine overdoses
Cyanide antidote package: sodium nitrate, sodium thoisulfate, amyl nitrate
Glucagon: antidote for Beta-adrenoreceptor blockers, may reverse low BP, bradycardia
Ethanol: used in ethylene glycol poisoning to decrease kidney damage, in MeOH poisoning as well
Fomepizole: an alcohol dehydrogenase inhibitor used to treat MeOH and ethylene glycol accidents
Diazepam: used for chemical-induced convulsions
Pyridoxine (Vitamin B6): used for isoniazid OD
Methylene blue: used to convert methemoglobin to hemoglobin (nitrate poisoning)
Know the following definitions:
Toxicology: relation of hazardous effects of chemicals, including drugs, to biological systems
Acute toxicity: adverse effect resulting from a single, usually large, exposure to a toxin
Subacute toxicity: Somewhat acute, between acute and chronic (online med. dictionary)
Chronic toxicity: harmful effect from repeated exposures to a toxin for 3+ months
Therapeutic index: LD50/ED50
Risk: probability that injury will result from exposure to a substance for given conditions, dose, and route
Threshold limit values (TLV): maximum safe ambient air concentrations of chemicals during a workweek,a 15 minute interval, and instantaneously
Know the following general mechanisms by which drugs or chemicals can cause toxicity:
Alterations in receptor-ligand interactions
Nicotine, most drugs
Alterations in membrane function
Local anesthetics, hydrocarbons
Interference with cellular energy mechanisms
Cyanide, Pentachlorophenol
Covalent binding to biomolecules
Organophosphates, alkylating agents
Interference with calcium homeostasis
Oxalates
By causing non-lethal alterations in somatic cells
Carcinogens, e.g. aflatoxin
Alterations in ligand-activated transcription factors
Dioxins
By inducing programmed cell death (apoptosis)
Acetaminophen
· Understand the primary determinants of toxicity
Dose and dose rate
Duration of exposure: long duration: bad.
(Principle) routes of exposure: inhalation, transdermal, and oral
· Understand how the following factors modify toxicity
Biotransformation
Parathion à Paraxon
Methanol à Formaldehyde à Formic Acid (MeOH shares pathway with EtOH)
Immune function
Review the hypersensitivities, e.g. Mosby 1-9 or perhaps even your long term memory
Phototoxicity: drug intermediates accumulate in skin, exposed to UV à toxic compounds
Examples: tetracycline, sulfonamides
Age: pharmodynamics, pharmokinetics vary over a lifetime
Gender: no specifics were discussed
· Understand how to manage a poisoned patient - know the ABCD's
Airway: should be cleared of vomitus or other obstruction and an airway or ET tube inserted
Breathing:Assesed by observation and measurements of arterial gases
Intubate and mechanically ventilate if necessary
Circulation
Monitor pulse rate, blood pressure, and urinary output
Start IV and draw blood for glucose and other labs
Dextrose: to every patient with altered mental status (thinking about hypoglycemic problems)
100mg thiamine to alcoholic and malnourished to prevent Wernicke-Korsakoff’s
· Understand the importance of the history and physical examination in treating a poisoned patient
Oral history may be unreliable for a number of reasons
Be on the lookout for classical OD effects of common meds and poisons
Treat the patient, not the poison
· Understand the importance of the following laboratory analysis and procedures
Arterial blood gases
CO2: increased with hypoventilation
PO2: low with aspiration pneumonia or drug-induced pulmonary edema
Reduced with poor tissue oxygenation due to hypoxia, hypotension, or Cn poisoning
May appear normal in CO because dissolved O2 is measured, not oxyhemoglobin
Electrolyte analysis (an ion gap)
(Na+ + K+) + (HCO3
- + Cl-) (this is just major cations minus anions)
normal = 12 plus/minus 4
elevated by:
renal failure
diabetic ketoacidosis
shock-induced lactic adicosis
Drug-induced metabolic acidosis (salicylates, methanol, ethylene glycol, isoniazid, Fe)
Renal and liver function tests
Renal tests: In the UA look for:
BUN, creatinine (nitrogen load and glomerular filtration checks, respectively)
CK and myoglobin (muscle insults)
oxlate crystals: suggest ethylene glycol poisoning
Liver tests: transaminases, look at the PT
Osmolar gap: useful in alcohols poisonings; alcohols will increase the osmolar gap (K. table 59-4)
Calculated serum osmolality: [ 2Na+ + glucose/18 + BUN/3]
Osmolar gap: measured osmolality – calculated osmolality
EKG examples:
Wide QRS: tricyclics, quinidine
Long QT: quinidine, phenothiazine, tricyclics
Variable AV block, screwy rhythms: dig. overdose
Ischemic changes: hypoxemia due to CO
· Understand how toxins can be removed or elimination can be enhanced
Gastric lavage
> 30 minutes have passed since the ingestion of a corrosive material
ingestion of hydrocarbons
coma, stupor, delerium, unconsciousness, convulsions
Induced emesis (know contraindications)
Syrup of Ipecac is often used; same contraindications as above
Increased rate of excretion
Catharsis: increased clearance of intestinal contents
Sorbitol is the preferred agent (hellooooo, apple juice), MgSO4 can be used if kidneys are ok
osmotic diuretics: e.g. mannitol, urea, etc.
Altered urinary pH
Alkalination: useful for salicylate or phenobarbitol overdose
Acidification: not recommended b/c worsens renal effects of rhabdomyolysis
Peritoneal dialysis: simple and available, but inefficient for most drugs
Hemodialysis: especially useful in cases where electrolyte and fluid imbalances are present
Hemoperfusion: blood pumped from pt’s vein through a cartridge filled with adsorbent material
Especially effective for high molecular weight compounds
· Understand the importance of the following specific drugs in managing the poisoned patient
Activated charcoal: large surface area, suggested dose is 10:1 charcoal to est. weight of toxin
Good for: adsorption of many drugs and poisons
No good for: Fe, Li, K, Cn, alcohols, corrosive acids and alkali, methylcarbmate, tolbutamide
Ipecac syrup: emetic agent; use 30 ml for adults, 10-15 ml for kids, repeated q 15 minutes if necessary
Emetic contraindications still apply: not for corrosives, hydrocarbons, rapidly acting convulsants
Ammonium chloride: used to acidify the urine
Not used much clinically because of side effects, e.g. indirect kidney damage
urinary excretion of weak organic bases
Sodium bicarbonate: urine alkalinization
urinary excretion of weak organic acids
Magnesium sulfate: cathartic, contraindicated in renal compromise
Mannitol: used to renal clearance of toxins (also to intraocular and intracranial pressure)
Deferoxamine: use IV or IM; chelator of choice for Fe poisoning
Dimercaprol: single-agent therapy for arsenic and mercury, use for lead with EDTA
Edetate, calcium disodium (EDTA): efficient chelator of many di/trivalent metal ions, esp. lead
Penicillamine: chelator of Cu and Pb
used in Wilson’s cystinuria, resistant cases of rheumatoid arthritis
Succimer (Dimercaptosuccinic Acid, DMSA), analog of dimercaprol,
prevents and reverses metal-induced inhibition of sulfhydryl-containing enzymes
urinary Pb excretion, protects against lethal effects of As, kidney [Hg]
Acetylcysteine: acetaminophen OD antidote, give within 8-10 hours of insult
Digoxin-specific FAb antibody: binds Dig
Atropine: used for cholinesterase poisonings to block ACh binding
Pralidoxime (2-PAM): cholinesterase reactivator, given only for organophosphates
Physostigmine: suggested for antimuscarinic, anticholinergic agents, but not for TCA’s
Flumazenil: used for benzodiazepine overdoses
Cyanide antidote package: sodium nitrate, sodium thoisulfate, amyl nitrate
Glucagon: antidote for Beta-adrenoreceptor blockers, may reverse low BP, bradycardia
Ethanol: used in ethylene glycol poisoning to decrease kidney damage, in MeOH poisoning as well
Fomepizole: an alcohol dehydrogenase inhibitor used to treat MeOH and ethylene glycol accidents
Diazepam: used for chemical-induced convulsions
Pyridoxine (Vitamin B6): used for isoniazid OD
Methylene blue: used to convert methemoglobin to hemoglobin (nitrate poisoning)
CHOLECYSTOKININ & Substance P
CHOLECYSTOKININ
-Cholecystokinin is produced in duodenum causing contraction of gall bladder.
-Popularly known as pancreozymin.
-When administered by IV injection causes increase in secretion of
Pancreatic enzymes and also stimulates gall bladder contraction.
-It is used as diagnostic aid for the purpose of testing pancreatic
function along with SECRETIN.
-It is also used as adjunct in cholecystography.
-Vasomotor reactions, abdominal discomfort and hypersensitivity has
been reported.
-Cholecystokinin is a Neuropeptide (octapeptide)
-cholecystokinin receptors are a group of G-protein coupled receptors.
-Antagonists to neuropeptide CCK proved to possess anxiolytic activity.
-*Loxiglumide is CCK antagonist.
SUBSTANCE.P
Substance .P is a neuropeptide .
It is a short chain with 11 amino acid polypeptide.
It functions as Neurotransmitter and neuromodulator.
It belongs to Tachykinin Family, Neuropeptide family.
In body it is majorily synthesized in CNS, bloodvessels, GIT, always and skin.
In CNS substance.P has been associated in regulation of mood
disorders, anxiety, stress, nausea, emesis, pain, respiratory rhythm,
neurogenesis and neurotoxicity.
Tachykinin Family includes NKA,NKB(NK-neurokinin) and Substance. P
*Pharmacological actions-
Action on Blood vessels-
It is a potent vasodilator.
This cause release of Nitric oxide from endothelium causing hypotension.
Action on Neurons-
Substance.P involved in transmission of pain impulses from peripheral
receptors to CNS.
Action on Vomiting center-
Vomiting center in brainstem (medulla oblongata).
Action on smooth muscle-
Causes contraction of smooth muscle and constriction of blood vessel.
*Substance P as a Pain Mediator-
It is a protein found in brain and spinal cord associated with
inflammatory process in joints.
-low back pain
- fibromyalgia
-Arthritis
Physiological Activities-
Vomiting reflex
Defensive behavior
Change in cardiovascular tone
Stimulation of salivary secret
Smooth contraction
Vasodilatation.
Receptors for Substance.P
The endogenous receptor for substance.p is NK-1, NK-1R.
These are G-protein receptors.
Treatment-
Capsaicin obtained from capsicum has been shown to reduce the levels
of substance p in nerve endings and decrease in pain by depletion of
c-fibre nerves.
Antagonist- Aprepitant
Structure- Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2.
-Cholecystokinin is produced in duodenum causing contraction of gall bladder.
-Popularly known as pancreozymin.
-When administered by IV injection causes increase in secretion of
Pancreatic enzymes and also stimulates gall bladder contraction.
-It is used as diagnostic aid for the purpose of testing pancreatic
function along with SECRETIN.
-It is also used as adjunct in cholecystography.
-Vasomotor reactions, abdominal discomfort and hypersensitivity has
been reported.
-Cholecystokinin is a Neuropeptide (octapeptide)
-cholecystokinin receptors are a group of G-protein coupled receptors.
-Antagonists to neuropeptide CCK proved to possess anxiolytic activity.
-*Loxiglumide is CCK antagonist.
SUBSTANCE.P
Substance .P is a neuropeptide .
It is a short chain with 11 amino acid polypeptide.
It functions as Neurotransmitter and neuromodulator.
It belongs to Tachykinin Family, Neuropeptide family.
In body it is majorily synthesized in CNS, bloodvessels, GIT, always and skin.
In CNS substance.P has been associated in regulation of mood
disorders, anxiety, stress, nausea, emesis, pain, respiratory rhythm,
neurogenesis and neurotoxicity.
Tachykinin Family includes NKA,NKB(NK-neurokinin) and Substance. P
*Pharmacological actions-
Action on Blood vessels-
It is a potent vasodilator.
This cause release of Nitric oxide from endothelium causing hypotension.
Action on Neurons-
Substance.P involved in transmission of pain impulses from peripheral
receptors to CNS.
Action on Vomiting center-
Vomiting center in brainstem (medulla oblongata).
Action on smooth muscle-
Causes contraction of smooth muscle and constriction of blood vessel.
*Substance P as a Pain Mediator-
It is a protein found in brain and spinal cord associated with
inflammatory process in joints.
-low back pain
- fibromyalgia
-Arthritis
Physiological Activities-
Vomiting reflex
Defensive behavior
Change in cardiovascular tone
Stimulation of salivary secret
Smooth contraction
Vasodilatation.
Receptors for Substance.P
The endogenous receptor for substance.p is NK-1, NK-1R.
These are G-protein receptors.
Treatment-
Capsaicin obtained from capsicum has been shown to reduce the levels
of substance p in nerve endings and decrease in pain by depletion of
c-fibre nerves.
Antagonist- Aprepitant
Structure- Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2.
PENTAGASTRIN
PENTAGASTRIN
Drug Category :
• Diagnostic aid
• gastric function
Indication: Used as a diagnostic aid for evaluation of gastric acid secretory function
Pharmacology: Pentagastrin is indicated as a diagnostic aid for evaluation of gastric acid secretory function. It is effective in testing for anacidity (achlorhydria) in patients with suspected pernicious anemia, atrophic gastritis, or gastric carcinoma. It is also effective in determining the reduction in acid output after operations for peptic ulcer, such as vagotomy or gastric resection.
Mechanism of Action: The exact mechanism by which pentagastrin stimulates gastric acid, pepsin, and intrinsic factor secretion is unknown; however, since pentagastrin is an analogue of natural gastrin, it is believed that it excites the oxyntic cells of the stomach to secrete to their maximum capacity. Pentagastrin stimulates pancreatic secretion, especially when administered in large intramuscular doses. Pentagastrin also increases gastrointestinal motility by a direct effect on the intestinal smooth muscle. However, it delays gastric emptying time probably by stimulation of terminal antral contractions, which enhance retropulsion.
Precautions-
It should be given with care to patients with
Acute peptic ulceration
Hepatic or biliary tract disease.
Adverse effects-
Pentgastrin may cause a no GIT effects. It causes Nausea and abdominal cramps.
Tachycardia
Arrhythmia
Dizziness, allergic reactions
Incidence less frequent or rare:- fast heartbeat ,usually transient chills, dizziness, faintness, or lightheadedness, drowsiness, feeling of heaviness of arms and legs, headache, increased sweating, numbness, tingling, pain, or weakness in hands or feet, shortness of breath, unusual tiredness
DOSAGE-
6 micrograms/kg by SC injection, IM or Nasal Inhalation.
Drug Category :
• Diagnostic aid
• gastric function
Indication: Used as a diagnostic aid for evaluation of gastric acid secretory function
Pharmacology: Pentagastrin is indicated as a diagnostic aid for evaluation of gastric acid secretory function. It is effective in testing for anacidity (achlorhydria) in patients with suspected pernicious anemia, atrophic gastritis, or gastric carcinoma. It is also effective in determining the reduction in acid output after operations for peptic ulcer, such as vagotomy or gastric resection.
Mechanism of Action: The exact mechanism by which pentagastrin stimulates gastric acid, pepsin, and intrinsic factor secretion is unknown; however, since pentagastrin is an analogue of natural gastrin, it is believed that it excites the oxyntic cells of the stomach to secrete to their maximum capacity. Pentagastrin stimulates pancreatic secretion, especially when administered in large intramuscular doses. Pentagastrin also increases gastrointestinal motility by a direct effect on the intestinal smooth muscle. However, it delays gastric emptying time probably by stimulation of terminal antral contractions, which enhance retropulsion.
Precautions-
It should be given with care to patients with
Acute peptic ulceration
Hepatic or biliary tract disease.
Adverse effects-
Pentgastrin may cause a no GIT effects. It causes Nausea and abdominal cramps.
Tachycardia
Arrhythmia
Dizziness, allergic reactions
Incidence less frequent or rare:- fast heartbeat ,usually transient chills, dizziness, faintness, or lightheadedness, drowsiness, feeling of heaviness of arms and legs, headache, increased sweating, numbness, tingling, pain, or weakness in hands or feet, shortness of breath, unusual tiredness
DOSAGE-
6 micrograms/kg by SC injection, IM or Nasal Inhalation.
ESTROGENS
ESTROGENS
Functions
While estrogens are present in both men and women, they are usually present at significantly higher levels in women of reproductive age. They promote the development of female secondary sex characteristics, such as breasts, and are also involved in the thickening of the endometrium and other aspects of regulating the menstrual cycle. In males, estrogen regulates certain functions of the reproductive system important to the maturation of sperm and may be necessary for a healthy libido. Furthermore, there are several other structural changes induced by estrogen, in addition to other functions.
• Structural
o promote formation of female secondary sex characteristics
o decelerate height growth[8]
o accelerate metabolism (burn fat)
o reduce muscle mass
o stimulate endometrial growth
o increase uterine growth
o maintenance of vessel and skin
o reduce bone resorption, increase bone formation
o morphic change (endomorphic -> mesomorphic -> ectomorphic)
• protein synthesis
o increase hepatic production of binding proteins
• coagulation
o increase circulating level of factors 2, 7, 9, 10, antithrombin III, plasminogen
o increase platelet adhesiveness
• Lipid
o increase HDL, triglyceride, height growth
o decrease LDL, fat depositition
• Fluid balance
o salt (sodium) and water retention
o increase growth hormone
o increase cortisol, SHBG
• gastrointestinal tract
o reduce bowel motility
o increase cholesterol in bile
• Melanin
o increase pheomelanin, reduce eumelanin
• Cancer
o support hormone-sensitive breast cancers (see section below)
• Lung function
o promotes lung function by supporting alveoli (in rodents but probably in humans).
Sexual desire is dependent on androgen levels rather than estrogen levels.
Role in cancer
About 80% of breast cancers, once established, rely on supplies of the hormone estrogen to grow: they are known as hormone-sensitive or hormone-receptor-positive cancers.Suppression of production in the body of estrogen is a treatment for these cancers.
Medical applications
Since estrogen circulating in the blood can negatively feed-back to reduce circulating levels of FSH and LH, most oral contraceptives contain a synthetic estrogen, along with a synthetic progestin. Even in men, the major hormone involved in LH feedback is estradiol, not testosterone.
As more fully discussed in the article on Hormone replacement therapy, estrogen and other hormones are given to postmenopausal women in order to prevent osteoporosis as well as treat the symptoms of menopause such as hot flashes, vaginal dryness, urinary stress incontinence, chilly sensations, dizziness, fatigue, irritability, and sweating. Fractures of the spine, wrist, and hips decrease by 50-70% and spinal bone density increases by ~5% in those women treated with estrogen within 3 years of the onset of menopause and for 5-10 years thereafter.
Before the specific dangers of conjugated equine estrogens were well understood, standard therapy was 0.625 mg/day of conjugated equine estrogens (such as Premarin). There are, however, risks associated with conjugated equine estrogen therapy. Among the older postmenopausal women studied as part of the Women's Health Initiative (WHI), an orally-administered conjugated equine estrogen supplement was found to be associated with an increased risk of dangerous blood clotting. The WHI studies used one type of estrogen supplement, a high oral dose of conjugated equine estrogens (Premarin alone and with medroxyprogesterone acetate as PremPro).
In a study by the NIH, esterified estrogens were not proven to pose the same risks to health as conjugated equine estrogens. Hormone replacement therapy has favorable effects on serum cholesterol levels, and when initiated immediately upon menopause reduces the incidence of cardiovascular disease. Estrogen has a protector effect on atherosclerosis : it lowers LDL and triglycerides, it raises HDL levels and has endothelial vasodilatation properties plus an anti-inflammatory component.
Research is underway to determine if risks of estrogen supplement use are the same for all methods of delivery. In particular, estrogen applied topically may have a different spectrum of side-effects than when administered orally, and transdermal oestrogens do not affect clotting as they are absorbed directly into the systemic circulation, avoiding first-pass metabolism in the liver. This route of administration is thus preferred in women with a history of thrombo-embolic disease.
Estrogen is also used in the therapy of vaginal atrophy, hypoestrogenism (as a result of hypogonadism, castration, or primary ovarian failure), amenorrhea, dysmenorrhea, and oligomenorrhea. Estrogens can also be used to suppress lactation after child birth.
Hormone-receptor-positive breast cancers are treated with drugs which suppress production in the body of estrogen. This technique, in the context of treatment of breast cancer, is known variously as hormonal therapy, hormone therapy, or anti-estrogen therapy (not to be confused with hormone replacement therapy). Certain foods such as soy may also suppress the proliferative effects of estrogen and are used as an alternative to hormone therapy.
In humans and mice, estrogen promotes wound healing.
At one time, estrogen was used to induce growth attenuation in tall girls.
Recently, estrogen-induced growth attenuation was used as part of the controversial Ashley Treatment to keep a developmentally disabled girl from growing to adult size.
Under certain circumstances, estrogen may also be used in males for treatment of prostate cancer.
Most recently, estrogen has been used in experimental research as a way to treat patients suffering from bulimia nervosa, in addition to Cognitive Behavioral Therapy, which is the established standard for treatment in bulimia cases. The estrogen research hypothesizes that the disease may be linked to a hormonal imbalance in the brain.
Estrogen has also been used in studies which indicate that it may be an effective drug for use in the treatment of traumatic liver injury.[19
Functions
While estrogens are present in both men and women, they are usually present at significantly higher levels in women of reproductive age. They promote the development of female secondary sex characteristics, such as breasts, and are also involved in the thickening of the endometrium and other aspects of regulating the menstrual cycle. In males, estrogen regulates certain functions of the reproductive system important to the maturation of sperm and may be necessary for a healthy libido. Furthermore, there are several other structural changes induced by estrogen, in addition to other functions.
• Structural
o promote formation of female secondary sex characteristics
o decelerate height growth[8]
o accelerate metabolism (burn fat)
o reduce muscle mass
o stimulate endometrial growth
o increase uterine growth
o maintenance of vessel and skin
o reduce bone resorption, increase bone formation
o morphic change (endomorphic -> mesomorphic -> ectomorphic)
• protein synthesis
o increase hepatic production of binding proteins
• coagulation
o increase circulating level of factors 2, 7, 9, 10, antithrombin III, plasminogen
o increase platelet adhesiveness
• Lipid
o increase HDL, triglyceride, height growth
o decrease LDL, fat depositition
• Fluid balance
o salt (sodium) and water retention
o increase growth hormone
o increase cortisol, SHBG
• gastrointestinal tract
o reduce bowel motility
o increase cholesterol in bile
• Melanin
o increase pheomelanin, reduce eumelanin
• Cancer
o support hormone-sensitive breast cancers (see section below)
• Lung function
o promotes lung function by supporting alveoli (in rodents but probably in humans).
Sexual desire is dependent on androgen levels rather than estrogen levels.
Role in cancer
About 80% of breast cancers, once established, rely on supplies of the hormone estrogen to grow: they are known as hormone-sensitive or hormone-receptor-positive cancers.Suppression of production in the body of estrogen is a treatment for these cancers.
Medical applications
Since estrogen circulating in the blood can negatively feed-back to reduce circulating levels of FSH and LH, most oral contraceptives contain a synthetic estrogen, along with a synthetic progestin. Even in men, the major hormone involved in LH feedback is estradiol, not testosterone.
As more fully discussed in the article on Hormone replacement therapy, estrogen and other hormones are given to postmenopausal women in order to prevent osteoporosis as well as treat the symptoms of menopause such as hot flashes, vaginal dryness, urinary stress incontinence, chilly sensations, dizziness, fatigue, irritability, and sweating. Fractures of the spine, wrist, and hips decrease by 50-70% and spinal bone density increases by ~5% in those women treated with estrogen within 3 years of the onset of menopause and for 5-10 years thereafter.
Before the specific dangers of conjugated equine estrogens were well understood, standard therapy was 0.625 mg/day of conjugated equine estrogens (such as Premarin). There are, however, risks associated with conjugated equine estrogen therapy. Among the older postmenopausal women studied as part of the Women's Health Initiative (WHI), an orally-administered conjugated equine estrogen supplement was found to be associated with an increased risk of dangerous blood clotting. The WHI studies used one type of estrogen supplement, a high oral dose of conjugated equine estrogens (Premarin alone and with medroxyprogesterone acetate as PremPro).
In a study by the NIH, esterified estrogens were not proven to pose the same risks to health as conjugated equine estrogens. Hormone replacement therapy has favorable effects on serum cholesterol levels, and when initiated immediately upon menopause reduces the incidence of cardiovascular disease. Estrogen has a protector effect on atherosclerosis : it lowers LDL and triglycerides, it raises HDL levels and has endothelial vasodilatation properties plus an anti-inflammatory component.
Research is underway to determine if risks of estrogen supplement use are the same for all methods of delivery. In particular, estrogen applied topically may have a different spectrum of side-effects than when administered orally, and transdermal oestrogens do not affect clotting as they are absorbed directly into the systemic circulation, avoiding first-pass metabolism in the liver. This route of administration is thus preferred in women with a history of thrombo-embolic disease.
Estrogen is also used in the therapy of vaginal atrophy, hypoestrogenism (as a result of hypogonadism, castration, or primary ovarian failure), amenorrhea, dysmenorrhea, and oligomenorrhea. Estrogens can also be used to suppress lactation after child birth.
Hormone-receptor-positive breast cancers are treated with drugs which suppress production in the body of estrogen. This technique, in the context of treatment of breast cancer, is known variously as hormonal therapy, hormone therapy, or anti-estrogen therapy (not to be confused with hormone replacement therapy). Certain foods such as soy may also suppress the proliferative effects of estrogen and are used as an alternative to hormone therapy.
In humans and mice, estrogen promotes wound healing.
At one time, estrogen was used to induce growth attenuation in tall girls.
Recently, estrogen-induced growth attenuation was used as part of the controversial Ashley Treatment to keep a developmentally disabled girl from growing to adult size.
Under certain circumstances, estrogen may also be used in males for treatment of prostate cancer.
Most recently, estrogen has been used in experimental research as a way to treat patients suffering from bulimia nervosa, in addition to Cognitive Behavioral Therapy, which is the established standard for treatment in bulimia cases. The estrogen research hypothesizes that the disease may be linked to a hormonal imbalance in the brain.
Estrogen has also been used in studies which indicate that it may be an effective drug for use in the treatment of traumatic liver injury.[19
Principles of Chemotherapy
CHEMOTHERAPY
Principles
Cancer is the uncontrolled growth of cells coupled with malignant behavior: invasion and metastasis. Cancer is thought to be caused by the interaction between genetic susceptibility and environmental toxins.
Broadly, most chemotherapeutic drugs work by impairing mitosis (cell division), effectively targeting fast-dividing cells. As these drugs cause damage to cells they are termed cytotoxic. Some drugs cause cells to undergo apoptosis (so-called "programmed cell death").
Unfortunately, scientists have yet to identify specific features of malignant and immune cells that would make them uniquely targetable (barring some recent examples, such as the Philadelphia chromosome as targeted by imatinib). This means that other fast dividing cells such as those responsible for hair growth and for replacement of the intestinal epithelium (lining) are also often affected. However, some drugs have a better side-effect profile than others, enabling doctors to adjust treatment regimens to the advantage of patients in certain situations.
As chemotherapy affects cell division, tumors with high growth fractions (such as acute myelogenous leukemia and the aggressive lymphomas, including Hodgkin's disease) are more sensitive to chemotherapy, as a larger proportion of the targeted cells are undergoing cell division at any time. Malignancies with slower growth rates, such as indolent lymphomas, tend to respond to chemotherapy much more modestly.
Drugs affect "younger" tumors (i.e. more differentiated) more effectively, because mechanisms regulating cell growth are usually still preserved. With succeeding generations of tumor cells, differentiation is typically lost, growth becomes less regulated, and tumors become less responsive to most chemotherapeutic agents. Near the center of some solid tumors, cell division has effectively ceased, making them insensitive to chemotherapy. Another problem with solid tumors is the fact that the chemotherapeutic agent often does not reach the core of the tumor. Solutions to this problem include radiation therapy (both brachytherapy and teletherapy) and surgery.
Over time, cancer cells become more resistant to chemotherapy treatments. Recently, scientists have identified small pumps on the surface of cancer cells that actively move chemotherapy from inside the cell to the outside. Research on p-glycoprotein and other such chemotherapy efflux pumps, is currently ongoing. Medications to inhibit the function of p-glycoprotein are undergoing testing as of June, 2007 to enhance the efficacy of chemotherapy.
When chemotherapy drugs attack reproducing cells, they cannot tell the difference between reproducing cells of normal tissues (that are replacing worn-out normal cells) and cancer cells. The damage to normal cells can cause side effects. Each time chemotherapy is given, it involves trying to find a balance between destroying the cancer cells (in order to cure or control the disease) and sparing the normal cells (to lessen unwanted side effects).
Treatment schemes
There are a number of strategies in the administration of chemotherapeutic drugs used today. Chemotherapy may be given with a curative intent or it may aim to prolong life or to palliate symptoms.
Combined modality chemotherapy is the use of drugs with other cancer treatments, such as radiation therapy or surgery. Most cancers are now treated in this way.
Combination chemotherapy is a similar practice which involves treating a patient with a number of different drugs simultaneously. The drugs differ in their mechanism and side effects. The biggest advantage is minimizing the chances of resistance developing to any one agent.
In
All chemotherapy regimens require that the patient be capable of undergoing the treatment. Performance status is often used as a measure to determine whether a patient can receive chemotherapy, or whether dose reduction is required.
Principles
Cancer is the uncontrolled growth of cells coupled with malignant behavior: invasion and metastasis. Cancer is thought to be caused by the interaction between genetic susceptibility and environmental toxins.
Broadly, most chemotherapeutic drugs work by impairing mitosis (cell division), effectively targeting fast-dividing cells. As these drugs cause damage to cells they are termed cytotoxic. Some drugs cause cells to undergo apoptosis (so-called "programmed cell death").
Unfortunately, scientists have yet to identify specific features of malignant and immune cells that would make them uniquely targetable (barring some recent examples, such as the Philadelphia chromosome as targeted by imatinib). This means that other fast dividing cells such as those responsible for hair growth and for replacement of the intestinal epithelium (lining) are also often affected. However, some drugs have a better side-effect profile than others, enabling doctors to adjust treatment regimens to the advantage of patients in certain situations.
As chemotherapy affects cell division, tumors with high growth fractions (such as acute myelogenous leukemia and the aggressive lymphomas, including Hodgkin's disease) are more sensitive to chemotherapy, as a larger proportion of the targeted cells are undergoing cell division at any time. Malignancies with slower growth rates, such as indolent lymphomas, tend to respond to chemotherapy much more modestly.
Drugs affect "younger" tumors (i.e. more differentiated) more effectively, because mechanisms regulating cell growth are usually still preserved. With succeeding generations of tumor cells, differentiation is typically lost, growth becomes less regulated, and tumors become less responsive to most chemotherapeutic agents. Near the center of some solid tumors, cell division has effectively ceased, making them insensitive to chemotherapy. Another problem with solid tumors is the fact that the chemotherapeutic agent often does not reach the core of the tumor. Solutions to this problem include radiation therapy (both brachytherapy and teletherapy) and surgery.
Over time, cancer cells become more resistant to chemotherapy treatments. Recently, scientists have identified small pumps on the surface of cancer cells that actively move chemotherapy from inside the cell to the outside. Research on p-glycoprotein and other such chemotherapy efflux pumps, is currently ongoing. Medications to inhibit the function of p-glycoprotein are undergoing testing as of June, 2007 to enhance the efficacy of chemotherapy.
When chemotherapy drugs attack reproducing cells, they cannot tell the difference between reproducing cells of normal tissues (that are replacing worn-out normal cells) and cancer cells. The damage to normal cells can cause side effects. Each time chemotherapy is given, it involves trying to find a balance between destroying the cancer cells (in order to cure or control the disease) and sparing the normal cells (to lessen unwanted side effects).
Treatment schemes
There are a number of strategies in the administration of chemotherapeutic drugs used today. Chemotherapy may be given with a curative intent or it may aim to prolong life or to palliate symptoms.
Combined modality chemotherapy is the use of drugs with other cancer treatments, such as radiation therapy or surgery. Most cancers are now treated in this way.
Combination chemotherapy is a similar practice which involves treating a patient with a number of different drugs simultaneously. The drugs differ in their mechanism and side effects. The biggest advantage is minimizing the chances of resistance developing to any one agent.
In
neoadjuvant chemotherapy(preoperative treatment) initial chemotherapy is aimed for shrinking the primary tumour, thereby rendering local therapy (surgery or radiotherapy) less destructive or more effective.
Adjuvant chemotherapy(postoperative treatment) can be used when there is little evidence of cancer present, but there is risk of recurrence. This can help reduce chances of resistance developing if the tumour does develop. It is also useful in killing any cancerous cells which have spread to other parts of the body. This is often effective as the newly growing tumours are fast-dividing, and therefore very susceptible.
Palliative chemotherapyis given without curative intent, but simply to decrease tumor load and increase life expectancy. For these regimens, a better toxicity profile is generally expected.
All chemotherapy regimens require that the patient be capable of undergoing the treatment. Performance status is often used as a measure to determine whether a patient can receive chemotherapy, or whether dose reduction is required.
Sunday, February 1, 2009
Quality audit
Quality Audit
Quality audit is the process of systematic examination of a quality system carried out by an internal or external quality auditor or an audit team [1]. It is an important part of organization's quality management system and is a key element in the ISO quality system standard, ISO 9001.
Quality audits are typically performed at predefined time intervals and ensure that the institution has clearly-defined internal quality monitoring procedures linked to effective action. This can help determine if the organization complies with the defined quality system processes and can involve procedural or results-based assessment criteria.
With the upgrade of the ISO9000 series of standards from the 1994 to 2000 series, the focus of the audits has shifted from purely procedural adherence towards measurement of the actual effectiveness of the Quality Management System (QMS) and the results that have been achieved through the implementation of a QMS.
Audits are an essential management tool to be used for verifying objective evidence of processes, to assess how successfully processes have been implemented, for judging the effectiveness of achieving any defined target levels, to provide evidence concerning reduction and elimination of problem areas. For the benefit of the organisation, quality auditing should not only report non-conformances and corrective actions, but also highlight areas of good practice. In this way other departments may share information and amend their working practices as a result, also contributing to continual improvement.
Quality audits can be an integral part of compliance or regulatory requirements.
Definition
Periodic, independent, and documented examination and verification of activities, records, processes, and other elements of a quality system to determine their conformity with the requirements of a quality standard such as ISO 9000. Any failure in their proper implementation may be published publicly and may lead to a revocation of quality certification. Also called conformity assessment or quality system audit.
Audits can also be used for safety purposes. Evans & Parker (2008) describe auditing as one of the most powerful safety monitoring techniques and 'an effective way to avoid complacency and highlight slowly deteriorating conditions', especially when the auditing focuses not just on compliance but effectiveness. [4]
The processes and tasks that a quality audit involves can be managed using a wide variety of software and self-assessment tools. Some of these relate specifically to quality in terms of fitness for purpose and conformance to standards, while others relate to Quality costs or, more accurately, to the Cost of poor quality. In analyzing quality costs, a cost of quality audit can be applied across any organization rather than just to conventional production or assembly processes
The principle behind Quality Audit
The principles of Quality Audit, in the sense we mean it here, are based on the style of quality standards used in several formal national and international standards such as the ISO-900x international quality standards. These standards do not in themselves create quality. The logic is as follows.
Every organisation should define comprehensive procedures by which their products or services can be delivered consistently to the desired level of quality. As was discussed in the section on Quality Management, maximum quality is rarely the desired objective since it can cost too much and take too long. The average product or service provides a sensible compromise between quality and cost. There is also a legitimate market for products that are low cost and low quality.
Standards authorities do not seek to make that business judgment and enforce it upon businesses, except where certain minimum standards must be met (eg all cars must have seat belts that meet minimum safety standards, but there is no attempt to define how elegant or comfortable they are).
The principle is that each organisation should create thorough, controlled procedures for each of its processes. Those procedures should deliver the quality that is sought. The Quality Audit, therefore, only needs to ensure that procedures have been defined, controlled, communicated and used. Processes will be put in place to deal with corrective actions when deviations occur. This principle can be applied to continuous business process operations or recurring project work. It would not be normal to establish a set of quality controlled procedures for a one-off situation since the emphasis is consistency.
This principle may be applied whether or not the organisation seeks to establish or maintain an externally recognised quality certification such as ISO-900x. To achieve a certification, the procedures will be subjected to internal and external scrutiny.
Preparing for Quality Audit
Thorough procedures need to be defined, controlled, communicated and used.
Thorough Procedures should cover all aspects of work where conformity and standards are required to achieved desired quality levels. For example, one might decide to control formal program testing, but leave the preliminary testing of a prototype to the programmer's discretion.
Procedures Any recurring aspect of work could merit regulation. The style and depth of the description will vary according to needs and preferences, provided it is sufficiently clear to be followed.
Defined A major tenet is that the defined procedures are good and will lead to the desired levels of quality. Considerable thought, consultation and trialing should be applied in order to define appropriate procedures. Procedures will often also require defined forms or software tools.
Controlled As with any good quality management, the procedures should be properly controlled in terms of accessibility, version control, update authorities etc.
Communicated All participants need to know about the defined procedures - that they exist, where to find them, what they cover. Quality reviewers are likely to check that team members understand about the procedures.
Used The defined procedures should be followed. Checks will be made to ensure this is the case. A corrective action procedure will be applied to deal with shortcomings. Typically the corrective action would either be to learn the lesson for next time, or to re-work the item if it is sufficiently important.
There is no reason why these Quality Audit techniques should conflict with the project's Quality Management processes. Where project work is recurring, the aim should be for the Quality Methods and other procedures to be defined once for both purposes.
Problems may occur where the current project has significant differences from earlier ones. Quality standards may have been set in stone as part of a quality certification. In extreme situations this can lead to wholly inappropriate procedures being forced upon the team, for example, using traditional structured analysis and design in a waterfall style approach for what would be handled best using iterative prototyping. The Project Manager may need to re-negotiate quality standards with the organization’s Quality Manager.
Operating Quality Audit
A Quality Audit approach affects the entire work lifecycle:
• Pre-defined standards will impact the way the project is planned
• Quality requirements for specific work packages and deliverables will be identified in advance
• Specific procedures will be followed at all stages
• Quality Methods must be defined and followed
• Completed work and deliverables should be reviewed for compliance.
This should be seen as an underlying framework and set of rules to apply in the project's Quality Management processes.
Quality audit is the process of systematic examination of a quality system carried out by an internal or external quality auditor or an audit team [1]. It is an important part of organization's quality management system and is a key element in the ISO quality system standard, ISO 9001.
Quality audits are typically performed at predefined time intervals and ensure that the institution has clearly-defined internal quality monitoring procedures linked to effective action. This can help determine if the organization complies with the defined quality system processes and can involve procedural or results-based assessment criteria.
With the upgrade of the ISO9000 series of standards from the 1994 to 2000 series, the focus of the audits has shifted from purely procedural adherence towards measurement of the actual effectiveness of the Quality Management System (QMS) and the results that have been achieved through the implementation of a QMS.
Audits are an essential management tool to be used for verifying objective evidence of processes, to assess how successfully processes have been implemented, for judging the effectiveness of achieving any defined target levels, to provide evidence concerning reduction and elimination of problem areas. For the benefit of the organisation, quality auditing should not only report non-conformances and corrective actions, but also highlight areas of good practice. In this way other departments may share information and amend their working practices as a result, also contributing to continual improvement.
Quality audits can be an integral part of compliance or regulatory requirements.
Definition
Periodic, independent, and documented examination and verification of activities, records, processes, and other elements of a quality system to determine their conformity with the requirements of a quality standard such as ISO 9000. Any failure in their proper implementation may be published publicly and may lead to a revocation of quality certification. Also called conformity assessment or quality system audit.
Audits can also be used for safety purposes. Evans & Parker (2008) describe auditing as one of the most powerful safety monitoring techniques and 'an effective way to avoid complacency and highlight slowly deteriorating conditions', especially when the auditing focuses not just on compliance but effectiveness. [4]
The processes and tasks that a quality audit involves can be managed using a wide variety of software and self-assessment tools. Some of these relate specifically to quality in terms of fitness for purpose and conformance to standards, while others relate to Quality costs or, more accurately, to the Cost of poor quality. In analyzing quality costs, a cost of quality audit can be applied across any organization rather than just to conventional production or assembly processes
The principle behind Quality Audit
The principles of Quality Audit, in the sense we mean it here, are based on the style of quality standards used in several formal national and international standards such as the ISO-900x international quality standards. These standards do not in themselves create quality. The logic is as follows.
Every organisation should define comprehensive procedures by which their products or services can be delivered consistently to the desired level of quality. As was discussed in the section on Quality Management, maximum quality is rarely the desired objective since it can cost too much and take too long. The average product or service provides a sensible compromise between quality and cost. There is also a legitimate market for products that are low cost and low quality.
Standards authorities do not seek to make that business judgment and enforce it upon businesses, except where certain minimum standards must be met (eg all cars must have seat belts that meet minimum safety standards, but there is no attempt to define how elegant or comfortable they are).
The principle is that each organisation should create thorough, controlled procedures for each of its processes. Those procedures should deliver the quality that is sought. The Quality Audit, therefore, only needs to ensure that procedures have been defined, controlled, communicated and used. Processes will be put in place to deal with corrective actions when deviations occur. This principle can be applied to continuous business process operations or recurring project work. It would not be normal to establish a set of quality controlled procedures for a one-off situation since the emphasis is consistency.
This principle may be applied whether or not the organisation seeks to establish or maintain an externally recognised quality certification such as ISO-900x. To achieve a certification, the procedures will be subjected to internal and external scrutiny.
Preparing for Quality Audit
Thorough procedures need to be defined, controlled, communicated and used.
Thorough Procedures should cover all aspects of work where conformity and standards are required to achieved desired quality levels. For example, one might decide to control formal program testing, but leave the preliminary testing of a prototype to the programmer's discretion.
Procedures Any recurring aspect of work could merit regulation. The style and depth of the description will vary according to needs and preferences, provided it is sufficiently clear to be followed.
Defined A major tenet is that the defined procedures are good and will lead to the desired levels of quality. Considerable thought, consultation and trialing should be applied in order to define appropriate procedures. Procedures will often also require defined forms or software tools.
Controlled As with any good quality management, the procedures should be properly controlled in terms of accessibility, version control, update authorities etc.
Communicated All participants need to know about the defined procedures - that they exist, where to find them, what they cover. Quality reviewers are likely to check that team members understand about the procedures.
Used The defined procedures should be followed. Checks will be made to ensure this is the case. A corrective action procedure will be applied to deal with shortcomings. Typically the corrective action would either be to learn the lesson for next time, or to re-work the item if it is sufficiently important.
There is no reason why these Quality Audit techniques should conflict with the project's Quality Management processes. Where project work is recurring, the aim should be for the Quality Methods and other procedures to be defined once for both purposes.
Problems may occur where the current project has significant differences from earlier ones. Quality standards may have been set in stone as part of a quality certification. In extreme situations this can lead to wholly inappropriate procedures being forced upon the team, for example, using traditional structured analysis and design in a waterfall style approach for what would be handled best using iterative prototyping. The Project Manager may need to re-negotiate quality standards with the organization’s Quality Manager.
Operating Quality Audit
A Quality Audit approach affects the entire work lifecycle:
• Pre-defined standards will impact the way the project is planned
• Quality requirements for specific work packages and deliverables will be identified in advance
• Specific procedures will be followed at all stages
• Quality Methods must be defined and followed
• Completed work and deliverables should be reviewed for compliance.
This should be seen as an underlying framework and set of rules to apply in the project's Quality Management processes.
Intellectual Property Rights
Intellectual Property Rights
Introduction
The protection of intellectual property rights in India continues to be strengthened. There is a well-established statutory, administrative and judicial framework to safeguard rights, whether they relate to patents, trademarks, copyright or industrial designs. Well-known international trademarks have been protected in India even when they were not registered in India. The Indian Trademarks Law has been extended through court decisions to service marks in addition to trade marks for goods. Computer software companies have successfully curtailed piracy through court orders. Computer databases have been protected. The courts, under the doctrine of breach of confidentiality, accorded an extensive protection of trade secrets. Right to privacy, which is not protected even in all developed countries, has been recognized in India. At Annexure 1 is a note on the steps being taken by the Government of India to strengthen and modernize the intellectual property administration system, including patent information services, trademarks registration and patent offices in India.
With specific reference to the obligations under Article 70.8 and 70.9 of the TRIPS Agreement, where India was required to provide a means for receipt of applications for product patents for pharmaceuticals and agricultural chemicals and to grant exclusive marketing rights on fulfillment of certain conditions, the Embassy would like to state the following:
1. India has established a mailbox system through administrative instructions. More than 2200 applications have been filed in this mailbox. About 600 of these applications have been filed by US companies.
2. No Exclusive Marketing Rights (EMR) application has so far been filed.
3. In respect of Article 70.8, the dispute raised by US related only to the legal security of the patent applications filed in the mail box system established through administrative instructions;
4. In respect of Article 70.9, the US argues that India should have had the necessary legislation in place for grant of EMR with effect from 1.1.95. India's stand was that the obligation to grant EMR arises only after an application for EMR is filed after fulfilling all the prescribed conditions and that, therefore, India had flexibility with regard to the timing of legislation;
5. In view of (3) and (4) above, the points raised by the US relate to only how and when India should fulfill these obligations. Even the US has not claimed that India has not accepted these obligations.
1. Comments on the Submission of Pharmaceutical Research and Manufacturers Association
PhRMA has mentioned that India should adopt a patent law which offers immediate product patent protection for pharmaceuticals in line with the highest international standards, and offer protection for all products not yet available in the Indian market. It is submitted that this is a demand that goes beyond India's obligations under the TRIPS Agreement as India is availing the full transition period therein. The ten year transition period available for providing product patents to pharmaceutical products is within WTO rules and a unilateral examination on the part of USA should not be allowed to overrule this multilateral understanding. Regarding the provision of the mailbox and EMR facility, PhRMA is aware that the issue has been taken up in the WTO and India has committed itself to implementation of the recommendations of the panel/appellate body.
Regarding the lack of intellectual property protection in the pharmaceutical sector, it may be mentioned that India does provide for patents in the pharmaceutical sector. However, in terms of Section 5 of the Patents Act, the patents are restricted to the methods of process of manufacture and not to the substances themselves. In terms of the TRIPS Agreement, India has time till January 1, 2005 to extend the product patent protection to this area. Therefore, there is no reason why the US pharmaceutical industry should have reservations on this score.
As regards the drug pricing policy of India, PhRMA has alleged that the pharmaceutical industry is unable to attract investment and the research based pharmaceutical industry is either withdrawing from India or not expanding operations. It is submitted that after the commencement of the liberalization process in India i.e. from August 1991 to November 1997 a total of 249 approvals for foreign direct investment have been obtained in the drug and pharmaceutical sector. These investment approvals envisage a total investment of Rs.6220.27 million. It may be noted that the number of approvals has doubled in 1994-96 as compared to 1991-93. In respect of the PhRMA averment that drug prices are being fixed on outdated cost data, the Department of Chemicals and Petrochemicals, Government of India has stated that the pricing is done based on actual cost data at manufacturing plants. It may be mentioned that the Government of India has not accepted the offer in 'good faith' for abolition of drug price controls. As regards the interests of Indian patients mentioned by PhRMA, under the present socio-economic conditions prevailing in India, drug price controls are required, particularly when no social security or an effective insurance system is yet in place.
PhRMA has mentioned that foreign companies experience arbitrary pricing norms of the Bureau of Industrial Costs & Prices, arbitrary local FDA decisions, high (42%) import duties and complex import procedures. It is submitted that these have no real basis and that specific cases need to be pointed out. The BICP pricing norms are based on accepted principles of costing viz. Actual cost plus a reasonable return of 14% on net worth and 22% on capital employed. 4% is added if the production is undertaken from basic stage. Further, FDA decisions are based on provisions of the Drugs and Cosmetics Act and the rules made thereunder. In fact, the US FDA has stricter norms than those prevailing in India. With regard to import duties, for pharmaceutical products, they have been brought down from 40% (plus 2%) to 30% (plus 5%) in the 1997-98 budget. These tariff rates have declined and are within India's WTO bindings. As regards import procedures, very few pharmaceutical items require import licenses. Only Penicillin and its derivatives, 6APA, Tetracycline, Oxytetracycline, Vitamin B 1, B2, Rifampicin and its intermediates and Streptomycin are placed in the negative list, requiring import licenses. Nevertheless, anomalies, if any, in the price control order and measures for free pricing are continuously under consideration of the Government of India.
PhRMA has stated that Government regulations regarding equity holdings are stringent and non-transparent. This is incorrect. In fact, the thrust of the modifications effected in Drug Policy is on attracting more investment so as to ensure the availability of quality medicines at reasonable prices in the country. The delicensing, the simplified price control mechanism and allowing higher return on investments made for basic stage manufacture would ensure a strong base for indigenous manufacture, which in turn will enhance availability. Automatic approvals to foreign equity participation up to 5 1 % and treating such companies at par with wholly Indian companies would help in bringing in newer technologies and making available newer products. Thus, modifications have made the Drug Policy attractive for domestic as well as foreign investment so as to ensure availability of quality medicines at reasonable prices.
On the question of drug price liability there is now a three-member committee, headed by a retired Judge of High Court, which looks into the Drug Price Equalization Account (DPEA) cases. The committee makes recommendations after hearing all affected parties in the matter. The Government while taking decisions in respect to DPEA considers these recommendations.
Copyright Law and Related Issues
The IIPA has been appreciative of the Copyright Act of 1957 as amended in 1994. It has observed that overall, the new law is Trade Related Aspects of Intellectual Property (TRIPS) compatible from the stand point of substantive rights, except that the term of protection for performers should be increased from 25 to 50 years. IIPA has also observed that the criminal provisions in the Act are among the toughest in the world. It, however, suggested that the level of fines should be increased. At present the minimum fine for the first offence is Rs.50, 000 and the maximum Rs. 200,000. In dollar terms this comes to 1,400 and 5,500. The amount of fines, decided in 1994, on the basis of the per capita income in India are satisfactory, even if they should appear to be low when one considers them in US dollars and in the context of the US per capita income. For making the Copyright Act TRIPS-compatible, India has time till January 1, 2000, as per Article 65.2 of the TRIPS Agreement.
IIPA has also suggested that the lack of presumptions with respect to ownership and subsistence of rights, and the need to prove actual knowledge must be remedied in amendments to the Copyright Act to make the law fully TRIPs-compatible, and bring it into conformity with the WIPO Copyright Treaty and the Performances and Phonograms Treaty.
In regard to the presumption of ownership of copyright issue, it is observed that in 1996 the Supreme Court of India, in a major ruling aimed at curbing piracy in audio and video cassettes in the country, said that ownership evidence of the original work was not required to prove the charge of copyright violation. Thus, even if there is no explicit provision in the Act, the case law makes the presumption of ownership of copyright. In fact the IIPA has also referred to the Supreme Court ruling.
Justifying suggested amendments to the Copyright Act in regard to the need to prove actual knowledge for raising/confiscating equipment for piracy, the IIPA has stated, "while raids on cable systems have been successful to date, it has been necessary to catch the pirate in the act of broadcasting a particular film without a licence in order for the police to be able to seize the pirate's broadcasting equipment." In this connection it may be mentioned that, in Section 64(l) of the Copyright Act, it has been clearly provided as under:
"Any police officer not below the rank of a sub-inspector, may, if he is satisfied that an offence under Section 63 in respect of the infringement of copyright in any work has been, is being, or is likely to be committed, seize without warrant, all copies of the work, and all plates used for the purpose of making infringing copies of the work, wherever found, and all copies and plates so seized shall, as soon as practicable, be produced before a Magistrate"
It will thus be seen that law does not require that the police witness the act of infringement in order to seize the goods involved. The law only requires that the police officer be satisfied that an offence "in respect of the infringement of copyright in any work has been, is being, or is likely to be committed" for the purpose of seizing, without warrant, all copies of the work and the plates used for making the infringing copies. This can further be adduced from the fact that he has the power to seize an infringing work even when he is satisfied that an offence is likely to be committed, thereby ruling out the essentiality of witnessing use of the goods to be seized in an act of infringement. Thus, the powers vested with the police take care of not only the past and present cases of infringement but also future cases where there is a likelihood of infringement of copyright provisions.
The WIPO Copyright Treaty, 1996 and the WIPO Performances and Phonograms Treaty, 1996 have not yet come into force as the requisite number of countries have not yet ratified or acceded to the treaties. India has also not yet acceded to the treaties. Since the two treaties have not yet come into force, it is premature to discuss the compatibility of the Copyright Act with these treaties.
The IIPA has also observed that as cable-showing of pirated products has begun to diminish, there has been an increase in unauthorized showing of US films in student unions in universities. The Copyright Act provides an exception "to the performance, in the course of the activities of an education institution, of a cinematography film,...if the audience is limited to ... staff and students, the parents and guardians of the students and persons directly connected with the activities of the institution..." (Section 52 (1)). IIPA has suggested that his provision should be interpreted narrowly to make such showings an infringement. If not, the provision must be amended and narrowed to be Berne and TRIPS-compatible. It may be observed that the Indian Copyright Act is fully compatible with the Berne and the Berne provisions included in the TRIPS. As such, no amendment is needed in the Act. However, the enforcement agencies may ensure that the exemption provisions for educational purposes are not misused for copyright infringement.
The IIPA in their submissions have also stated, "on the recording side, RIAA is concerned about an apparently over board provision in section 33 which may limit the ability of authors, and composers, as well as producers, to licence their works individually, rather than through a registered collection society." IIPA has suggested that this should be clarified in favor of full rights of all right holders to licence their rights individually. Section 33 of the Copyright Act, which provides for copyright societies, does not take away the rights of individual copyright owners. As per proviso to Section 33 "an owner of copyright shall, in his individual capacity, continue to have the right to grant licences in respect of his own works consistent with his obligations as a member of the registered. copyright society."
IIPA have also demanded a narrower interpretation of provisions relating to educational use of copyrighted work. First of all, no such instance has been brought to the notice of the Department of Education, Ministry of Human Resource Development, Government of India, which is the nodal Ministry for copyright. Further, it is quite likely that instances, if any, do not involve any commercial losses.
Modernization of Intellectual Property Administration System
(Annexure 1)
Government of India has taken several measures to streamline and strengthen the intellectual property administration systems in the country. Two projects were recently implemented with the help of WIPO/UNDP for the modernization of patent information services and trademarks registry, details of which are given below:
Modernization of Patent Information Services
I . The Office of Patent Information System (PIS) was set up in 1980 with a view to provide scientific and technological information contained in patents documents to the users, namely scientists, researchers, universities, Indian Institutes of Technology and others for taking up further research and development. It is estimated that about 80% of the world's knowledge of viable and adaptable technology are contained in about 30 million patent documents. One million documents are being added every year, covering 400,000 inventions. If the information contained in these documents is made available to industry, it can provide information for further research, facilitate evaluation of technology to be purchased under licensing arrangements, and identify worldwide developments in specific fields.
2. It was, therefore, felt necessary to establish a modernized patent information service centre through upgrading and mechanization of procedures relating to collection, retrieval and dissemination of patent information, using state-of-the-art technology and trained personnel.
3. Accordingly, a project for modernization of PIS was taken up with the financial assistance of UPDP and Government of India (at a cost of US$1.22 million of which $0.69 million provided by LTNDP) and technical assistance of World Intellectual Property Organisation (WIPO) in January, 1992. This project has been completed in June 1996.
4. As a part of activities, several awareness/training programs were organized in India for the benefit of scientific and industrial communities. Additional manpower to PIS was provided along with modem office equipment such as computers, CD-ROMS and on-line access to external and internal databases. Computerized Indian patent databases have also been established. All these activities are expected to result in better and improved services to the users.
Modernization of Trade Marks Registry
1. The use and promotion of trademarks helps inform consumers about goods and services available in the market. The use of trademarks facilitates identifying those responsible for putting those goods in the market. An effective trademarks system would help protect consumers against misleading practices. Therefore, it was recognized that in the interest of consumers, the Government of India should promote the use and provide effective protection for trademarks.
2. Accordingly, a project for modernization of trademarks registry was taken up in April 1993 at a cost of US$1.24 million, out of which US$ 0.58 million were provided by the UNDP. The project has been implemented in May 1996 with the technical assistance of WIPO.
3. Under the project, the capability of the Registry has been enhanced by modernizing and streamlining trade marks registration procedures; posts of electronic data processing personnel have been created; awareness programs organized; Registry has been strengthened in terms of modem office equipment; networking established between Head and Branch Offices, etc. It is felt that it would now be possible to issue registration certificate in a shorter period than before.
4. Further strengthening of the capabilities of the trademark registry is also being taken up in the Ninth Plan and a provision has also been incorporated in the Annual Plan 1997-98.
Modernization of Patent Offices
1. A proposal is now under consideration for taking up modernization of Patent Offices.
2. The Patent Offices, apart from grant of patent, perform other functions like public information and guidance, maintenance of public library and search room, production of journals, publication of official gazette, etc.
3. In view of the economic initiatives and liberalization measures taken by the Government since mid-1991, the number of applications for patents are expected to increase significantly. From a level of 3552 in 1991-92, it has increased to 8562 in 1996-97 and is expected to rise further in coming years. Accordingly, a proposal is under consideration for modernization of patent offices incorporating the following elements:-
• Human Resource Development
• Infrastructure Support and Strengthening
• Computerization and Modernization
• Elimination of backlog of patent applications
4. The project will be implemented in a period of 5 years. A detailed project report has already been prepared and the project has been included in Ninth Five-Year Plan and Annual Plan 1998-99.
Introduction
The protection of intellectual property rights in India continues to be strengthened. There is a well-established statutory, administrative and judicial framework to safeguard rights, whether they relate to patents, trademarks, copyright or industrial designs. Well-known international trademarks have been protected in India even when they were not registered in India. The Indian Trademarks Law has been extended through court decisions to service marks in addition to trade marks for goods. Computer software companies have successfully curtailed piracy through court orders. Computer databases have been protected. The courts, under the doctrine of breach of confidentiality, accorded an extensive protection of trade secrets. Right to privacy, which is not protected even in all developed countries, has been recognized in India. At Annexure 1 is a note on the steps being taken by the Government of India to strengthen and modernize the intellectual property administration system, including patent information services, trademarks registration and patent offices in India.
With specific reference to the obligations under Article 70.8 and 70.9 of the TRIPS Agreement, where India was required to provide a means for receipt of applications for product patents for pharmaceuticals and agricultural chemicals and to grant exclusive marketing rights on fulfillment of certain conditions, the Embassy would like to state the following:
1. India has established a mailbox system through administrative instructions. More than 2200 applications have been filed in this mailbox. About 600 of these applications have been filed by US companies.
2. No Exclusive Marketing Rights (EMR) application has so far been filed.
3. In respect of Article 70.8, the dispute raised by US related only to the legal security of the patent applications filed in the mail box system established through administrative instructions;
4. In respect of Article 70.9, the US argues that India should have had the necessary legislation in place for grant of EMR with effect from 1.1.95. India's stand was that the obligation to grant EMR arises only after an application for EMR is filed after fulfilling all the prescribed conditions and that, therefore, India had flexibility with regard to the timing of legislation;
5. In view of (3) and (4) above, the points raised by the US relate to only how and when India should fulfill these obligations. Even the US has not claimed that India has not accepted these obligations.
1. Comments on the Submission of Pharmaceutical Research and Manufacturers Association
PhRMA has mentioned that India should adopt a patent law which offers immediate product patent protection for pharmaceuticals in line with the highest international standards, and offer protection for all products not yet available in the Indian market. It is submitted that this is a demand that goes beyond India's obligations under the TRIPS Agreement as India is availing the full transition period therein. The ten year transition period available for providing product patents to pharmaceutical products is within WTO rules and a unilateral examination on the part of USA should not be allowed to overrule this multilateral understanding. Regarding the provision of the mailbox and EMR facility, PhRMA is aware that the issue has been taken up in the WTO and India has committed itself to implementation of the recommendations of the panel/appellate body.
Regarding the lack of intellectual property protection in the pharmaceutical sector, it may be mentioned that India does provide for patents in the pharmaceutical sector. However, in terms of Section 5 of the Patents Act, the patents are restricted to the methods of process of manufacture and not to the substances themselves. In terms of the TRIPS Agreement, India has time till January 1, 2005 to extend the product patent protection to this area. Therefore, there is no reason why the US pharmaceutical industry should have reservations on this score.
As regards the drug pricing policy of India, PhRMA has alleged that the pharmaceutical industry is unable to attract investment and the research based pharmaceutical industry is either withdrawing from India or not expanding operations. It is submitted that after the commencement of the liberalization process in India i.e. from August 1991 to November 1997 a total of 249 approvals for foreign direct investment have been obtained in the drug and pharmaceutical sector. These investment approvals envisage a total investment of Rs.6220.27 million. It may be noted that the number of approvals has doubled in 1994-96 as compared to 1991-93. In respect of the PhRMA averment that drug prices are being fixed on outdated cost data, the Department of Chemicals and Petrochemicals, Government of India has stated that the pricing is done based on actual cost data at manufacturing plants. It may be mentioned that the Government of India has not accepted the offer in 'good faith' for abolition of drug price controls. As regards the interests of Indian patients mentioned by PhRMA, under the present socio-economic conditions prevailing in India, drug price controls are required, particularly when no social security or an effective insurance system is yet in place.
PhRMA has mentioned that foreign companies experience arbitrary pricing norms of the Bureau of Industrial Costs & Prices, arbitrary local FDA decisions, high (42%) import duties and complex import procedures. It is submitted that these have no real basis and that specific cases need to be pointed out. The BICP pricing norms are based on accepted principles of costing viz. Actual cost plus a reasonable return of 14% on net worth and 22% on capital employed. 4% is added if the production is undertaken from basic stage. Further, FDA decisions are based on provisions of the Drugs and Cosmetics Act and the rules made thereunder. In fact, the US FDA has stricter norms than those prevailing in India. With regard to import duties, for pharmaceutical products, they have been brought down from 40% (plus 2%) to 30% (plus 5%) in the 1997-98 budget. These tariff rates have declined and are within India's WTO bindings. As regards import procedures, very few pharmaceutical items require import licenses. Only Penicillin and its derivatives, 6APA, Tetracycline, Oxytetracycline, Vitamin B 1, B2, Rifampicin and its intermediates and Streptomycin are placed in the negative list, requiring import licenses. Nevertheless, anomalies, if any, in the price control order and measures for free pricing are continuously under consideration of the Government of India.
PhRMA has stated that Government regulations regarding equity holdings are stringent and non-transparent. This is incorrect. In fact, the thrust of the modifications effected in Drug Policy is on attracting more investment so as to ensure the availability of quality medicines at reasonable prices in the country. The delicensing, the simplified price control mechanism and allowing higher return on investments made for basic stage manufacture would ensure a strong base for indigenous manufacture, which in turn will enhance availability. Automatic approvals to foreign equity participation up to 5 1 % and treating such companies at par with wholly Indian companies would help in bringing in newer technologies and making available newer products. Thus, modifications have made the Drug Policy attractive for domestic as well as foreign investment so as to ensure availability of quality medicines at reasonable prices.
On the question of drug price liability there is now a three-member committee, headed by a retired Judge of High Court, which looks into the Drug Price Equalization Account (DPEA) cases. The committee makes recommendations after hearing all affected parties in the matter. The Government while taking decisions in respect to DPEA considers these recommendations.
Copyright Law and Related Issues
The IIPA has been appreciative of the Copyright Act of 1957 as amended in 1994. It has observed that overall, the new law is Trade Related Aspects of Intellectual Property (TRIPS) compatible from the stand point of substantive rights, except that the term of protection for performers should be increased from 25 to 50 years. IIPA has also observed that the criminal provisions in the Act are among the toughest in the world. It, however, suggested that the level of fines should be increased. At present the minimum fine for the first offence is Rs.50, 000 and the maximum Rs. 200,000. In dollar terms this comes to 1,400 and 5,500. The amount of fines, decided in 1994, on the basis of the per capita income in India are satisfactory, even if they should appear to be low when one considers them in US dollars and in the context of the US per capita income. For making the Copyright Act TRIPS-compatible, India has time till January 1, 2000, as per Article 65.2 of the TRIPS Agreement.
IIPA has also suggested that the lack of presumptions with respect to ownership and subsistence of rights, and the need to prove actual knowledge must be remedied in amendments to the Copyright Act to make the law fully TRIPs-compatible, and bring it into conformity with the WIPO Copyright Treaty and the Performances and Phonograms Treaty.
In regard to the presumption of ownership of copyright issue, it is observed that in 1996 the Supreme Court of India, in a major ruling aimed at curbing piracy in audio and video cassettes in the country, said that ownership evidence of the original work was not required to prove the charge of copyright violation. Thus, even if there is no explicit provision in the Act, the case law makes the presumption of ownership of copyright. In fact the IIPA has also referred to the Supreme Court ruling.
Justifying suggested amendments to the Copyright Act in regard to the need to prove actual knowledge for raising/confiscating equipment for piracy, the IIPA has stated, "while raids on cable systems have been successful to date, it has been necessary to catch the pirate in the act of broadcasting a particular film without a licence in order for the police to be able to seize the pirate's broadcasting equipment." In this connection it may be mentioned that, in Section 64(l) of the Copyright Act, it has been clearly provided as under:
"Any police officer not below the rank of a sub-inspector, may, if he is satisfied that an offence under Section 63 in respect of the infringement of copyright in any work has been, is being, or is likely to be committed, seize without warrant, all copies of the work, and all plates used for the purpose of making infringing copies of the work, wherever found, and all copies and plates so seized shall, as soon as practicable, be produced before a Magistrate"
It will thus be seen that law does not require that the police witness the act of infringement in order to seize the goods involved. The law only requires that the police officer be satisfied that an offence "in respect of the infringement of copyright in any work has been, is being, or is likely to be committed" for the purpose of seizing, without warrant, all copies of the work and the plates used for making the infringing copies. This can further be adduced from the fact that he has the power to seize an infringing work even when he is satisfied that an offence is likely to be committed, thereby ruling out the essentiality of witnessing use of the goods to be seized in an act of infringement. Thus, the powers vested with the police take care of not only the past and present cases of infringement but also future cases where there is a likelihood of infringement of copyright provisions.
The WIPO Copyright Treaty, 1996 and the WIPO Performances and Phonograms Treaty, 1996 have not yet come into force as the requisite number of countries have not yet ratified or acceded to the treaties. India has also not yet acceded to the treaties. Since the two treaties have not yet come into force, it is premature to discuss the compatibility of the Copyright Act with these treaties.
The IIPA has also observed that as cable-showing of pirated products has begun to diminish, there has been an increase in unauthorized showing of US films in student unions in universities. The Copyright Act provides an exception "to the performance, in the course of the activities of an education institution, of a cinematography film,...if the audience is limited to ... staff and students, the parents and guardians of the students and persons directly connected with the activities of the institution..." (Section 52 (1)). IIPA has suggested that his provision should be interpreted narrowly to make such showings an infringement. If not, the provision must be amended and narrowed to be Berne and TRIPS-compatible. It may be observed that the Indian Copyright Act is fully compatible with the Berne and the Berne provisions included in the TRIPS. As such, no amendment is needed in the Act. However, the enforcement agencies may ensure that the exemption provisions for educational purposes are not misused for copyright infringement.
The IIPA in their submissions have also stated, "on the recording side, RIAA is concerned about an apparently over board provision in section 33 which may limit the ability of authors, and composers, as well as producers, to licence their works individually, rather than through a registered collection society." IIPA has suggested that this should be clarified in favor of full rights of all right holders to licence their rights individually. Section 33 of the Copyright Act, which provides for copyright societies, does not take away the rights of individual copyright owners. As per proviso to Section 33 "an owner of copyright shall, in his individual capacity, continue to have the right to grant licences in respect of his own works consistent with his obligations as a member of the registered. copyright society."
IIPA have also demanded a narrower interpretation of provisions relating to educational use of copyrighted work. First of all, no such instance has been brought to the notice of the Department of Education, Ministry of Human Resource Development, Government of India, which is the nodal Ministry for copyright. Further, it is quite likely that instances, if any, do not involve any commercial losses.
Modernization of Intellectual Property Administration System
(Annexure 1)
Government of India has taken several measures to streamline and strengthen the intellectual property administration systems in the country. Two projects were recently implemented with the help of WIPO/UNDP for the modernization of patent information services and trademarks registry, details of which are given below:
Modernization of Patent Information Services
I . The Office of Patent Information System (PIS) was set up in 1980 with a view to provide scientific and technological information contained in patents documents to the users, namely scientists, researchers, universities, Indian Institutes of Technology and others for taking up further research and development. It is estimated that about 80% of the world's knowledge of viable and adaptable technology are contained in about 30 million patent documents. One million documents are being added every year, covering 400,000 inventions. If the information contained in these documents is made available to industry, it can provide information for further research, facilitate evaluation of technology to be purchased under licensing arrangements, and identify worldwide developments in specific fields.
2. It was, therefore, felt necessary to establish a modernized patent information service centre through upgrading and mechanization of procedures relating to collection, retrieval and dissemination of patent information, using state-of-the-art technology and trained personnel.
3. Accordingly, a project for modernization of PIS was taken up with the financial assistance of UPDP and Government of India (at a cost of US$1.22 million of which $0.69 million provided by LTNDP) and technical assistance of World Intellectual Property Organisation (WIPO) in January, 1992. This project has been completed in June 1996.
4. As a part of activities, several awareness/training programs were organized in India for the benefit of scientific and industrial communities. Additional manpower to PIS was provided along with modem office equipment such as computers, CD-ROMS and on-line access to external and internal databases. Computerized Indian patent databases have also been established. All these activities are expected to result in better and improved services to the users.
Modernization of Trade Marks Registry
1. The use and promotion of trademarks helps inform consumers about goods and services available in the market. The use of trademarks facilitates identifying those responsible for putting those goods in the market. An effective trademarks system would help protect consumers against misleading practices. Therefore, it was recognized that in the interest of consumers, the Government of India should promote the use and provide effective protection for trademarks.
2. Accordingly, a project for modernization of trademarks registry was taken up in April 1993 at a cost of US$1.24 million, out of which US$ 0.58 million were provided by the UNDP. The project has been implemented in May 1996 with the technical assistance of WIPO.
3. Under the project, the capability of the Registry has been enhanced by modernizing and streamlining trade marks registration procedures; posts of electronic data processing personnel have been created; awareness programs organized; Registry has been strengthened in terms of modem office equipment; networking established between Head and Branch Offices, etc. It is felt that it would now be possible to issue registration certificate in a shorter period than before.
4. Further strengthening of the capabilities of the trademark registry is also being taken up in the Ninth Plan and a provision has also been incorporated in the Annual Plan 1997-98.
Modernization of Patent Offices
1. A proposal is now under consideration for taking up modernization of Patent Offices.
2. The Patent Offices, apart from grant of patent, perform other functions like public information and guidance, maintenance of public library and search room, production of journals, publication of official gazette, etc.
3. In view of the economic initiatives and liberalization measures taken by the Government since mid-1991, the number of applications for patents are expected to increase significantly. From a level of 3552 in 1991-92, it has increased to 8562 in 1996-97 and is expected to rise further in coming years. Accordingly, a proposal is under consideration for modernization of patent offices incorporating the following elements:-
• Human Resource Development
• Infrastructure Support and Strengthening
• Computerization and Modernization
• Elimination of backlog of patent applications
4. The project will be implemented in a period of 5 years. A detailed project report has already been prepared and the project has been included in Ninth Five-Year Plan and Annual Plan 1998-99.
Types of process validation
Types of process validation
Depending on when it is performed in relation to production, validation can be prospective, concurrent, retrospective or revalidation (repeated validation).
Prospective validation
Itis carried out during the development stage by means of a risk analysis of the production process, which is broken down into individual steps: these are then evaluated on the basis of past experience to determine whether they might lead to critical situations.
Where possible critical situations are identified, the risk is evaluated, the potential causes are investigated and assessed for probability and extent, the trial plans are drawn up, and the priorities set. The trials are then performed and evaluated, and an overall assessment is made. If, at the end, the results are acceptable, the process is satisfactory. Unsatisfactory processes must be modified and improved until a validation exercise proves them to be satisfactory. This form of validation is essential in order to limit the risk of errors occurring on the production scale, e.g. in the preparation of injectable products.
Concurrent validation
is carried out during normal production. This method is effective only if the development stage has resulted in a proper understanding of the fundamentals of the process. The first three production-scale batches must be monitored as comprehensively as possible.1The nature and specifications of subsequent in-process and final tests are based on the evaluation of the results of such monitoring.
1 This careful monitoring of the first three production batches is sometimes regarded as prospective validation.
Concurrent validation together with a trend analysis including stability should be carried out to an appropriate extent throughout the life of the product.
Retrospective validation
involves the examination of past experience of production on the assumption that composition, procedures, and equipment remain unchanged; such experience and the results of in-process and final control tests are then evaluated. Recorded difficulties and failures in production are analysed to determine the limits of process parameters. A trend analysis may be conducted to determine the extent to which the process parameters are within the permissible range.
Retrospective validation is obviously not a quality assurance measure in itself, and should never be applied to new processes or products. It may be considered in special circumstances only, e.g. when validation requirements are first introduced in a company. Retrospective validation may then be useful in establishing the priorities for the validation programme. If the results of a retrospective validation are positive, this indicates that the process is not in need of immediate attention and may be validated in accordance with the normal schedule. For tablets which have been compressed under individual pressure-sensitive cells, and with qualified equipment, retrospective validation is the most comprehensive test of the overall manufacturing process of this dosage form. On the other hand, it should not be applied in the manufacture of sterile products.
Revalidation
is needed to ensure that changes in the process and/or in the process environment, whether intentional or unintentional, do not adversely affect process characteristics and product quality.
Revalidation may be divided into two broad categories:
• Revalidation after any change having a bearing on product quality.
• Periodic revalidation carried out at scheduled intervals.
Revalidation after changes. Revalidation must be performed on introduction of any changes affecting a manufacturing and/or standard procedure having a bearing on the established product performance characteristics. Such changes may include those in starting material, packaging material, manufacturing processes, equipment, in-process controls, manufacturing areas, or support systems (water, steam, etc.). Every such change requested should be reviewed by a qualified validation group, which will decide whether it is significant enough to justify revalidation and, if so, its extent.
Revalidation after changes may be based on the performance of the same tests and activities as those used during the original validation, including tests on subprocesses and on the equipment concerned. Some typical changes which require revalidation include the following:
• Changes in the starting material(s). Changes in the physical properties, such as density, viscosity, particle size distribution, and crystal type and modification, of the active ingredients or excipients may affect the mechanical properties of the material; as a consequence, they may adversely affect the process or the product.
• Changes in the packaging material, e.g. replacing plastics by glass, may require changes in the packaging procedure and therefore affect product stability.
• Changes in the process, e.g. changes in mixing time, drying temperature and cooling regime, may affect subsequent process steps and product quality.
• Changes in equipment, including measuring instruments, may affect both the process and the product; repair and maintenance work, such as the replacement of major equipment components, may affect the process.
• Changes in the production area and support system, e.g. the rearrangement of manufacturing areas and/or support systems, may result in changes in the process. The repair and maintenance of support systems, such as ventilation, may change the environmental conditions and, as a consequence, revalidation/requalification may be necessary, mainly in the manufacture of sterile products.
• Unexpected changes and deviations may be observed during self-inspection or audit, or during the continuous trend analysis of process data.
Periodic revalidation. It is well known that process changes may occur gradually even if experienced operators work correctly according to established methods. Similarly, equipment wear may also cause gradual changes. Consequently, revalidation at scheduled times is advisable even if no changes have been deliberately made.
The decision to introduce periodic revalidation should be based essentially on a review of historical data, i.e. data generated during in-process and finished product testing after the latest validation, aimed at verifying that the process is under control. During the review of such historical data, any trend in the data collected should be evaluated.
In some processes, such as sterilization, additional process testing is required to complement the historical data. The degree of testing required will be apparent from the original validation.
Additionally, the following points should be checked at the time of a scheduled revalidation:
• Have any changes in master formula and methods, batch size, etc., occurred? If so, has their impact on the product been assessed?
• Have calibrations been made in accordance with the established programme and time schedule?
• Has preventive maintenance been performed in accordance with the programme and time schedule?
• Have the standard operating procedures (SOPs) been properly updated?
• Have the SOPs been implemented?
• Have the cleaning and hygiene programmes been carried out?
• Have any changes been made in the analytical control methods?
Depending on when it is performed in relation to production, validation can be prospective, concurrent, retrospective or revalidation (repeated validation).
Prospective validation
Itis carried out during the development stage by means of a risk analysis of the production process, which is broken down into individual steps: these are then evaluated on the basis of past experience to determine whether they might lead to critical situations.
Where possible critical situations are identified, the risk is evaluated, the potential causes are investigated and assessed for probability and extent, the trial plans are drawn up, and the priorities set. The trials are then performed and evaluated, and an overall assessment is made. If, at the end, the results are acceptable, the process is satisfactory. Unsatisfactory processes must be modified and improved until a validation exercise proves them to be satisfactory. This form of validation is essential in order to limit the risk of errors occurring on the production scale, e.g. in the preparation of injectable products.
Concurrent validation
is carried out during normal production. This method is effective only if the development stage has resulted in a proper understanding of the fundamentals of the process. The first three production-scale batches must be monitored as comprehensively as possible.1The nature and specifications of subsequent in-process and final tests are based on the evaluation of the results of such monitoring.
1 This careful monitoring of the first three production batches is sometimes regarded as prospective validation.
Concurrent validation together with a trend analysis including stability should be carried out to an appropriate extent throughout the life of the product.
Retrospective validation
involves the examination of past experience of production on the assumption that composition, procedures, and equipment remain unchanged; such experience and the results of in-process and final control tests are then evaluated. Recorded difficulties and failures in production are analysed to determine the limits of process parameters. A trend analysis may be conducted to determine the extent to which the process parameters are within the permissible range.
Retrospective validation is obviously not a quality assurance measure in itself, and should never be applied to new processes or products. It may be considered in special circumstances only, e.g. when validation requirements are first introduced in a company. Retrospective validation may then be useful in establishing the priorities for the validation programme. If the results of a retrospective validation are positive, this indicates that the process is not in need of immediate attention and may be validated in accordance with the normal schedule. For tablets which have been compressed under individual pressure-sensitive cells, and with qualified equipment, retrospective validation is the most comprehensive test of the overall manufacturing process of this dosage form. On the other hand, it should not be applied in the manufacture of sterile products.
Revalidation
is needed to ensure that changes in the process and/or in the process environment, whether intentional or unintentional, do not adversely affect process characteristics and product quality.
Revalidation may be divided into two broad categories:
• Revalidation after any change having a bearing on product quality.
• Periodic revalidation carried out at scheduled intervals.
Revalidation after changes. Revalidation must be performed on introduction of any changes affecting a manufacturing and/or standard procedure having a bearing on the established product performance characteristics. Such changes may include those in starting material, packaging material, manufacturing processes, equipment, in-process controls, manufacturing areas, or support systems (water, steam, etc.). Every such change requested should be reviewed by a qualified validation group, which will decide whether it is significant enough to justify revalidation and, if so, its extent.
Revalidation after changes may be based on the performance of the same tests and activities as those used during the original validation, including tests on subprocesses and on the equipment concerned. Some typical changes which require revalidation include the following:
• Changes in the starting material(s). Changes in the physical properties, such as density, viscosity, particle size distribution, and crystal type and modification, of the active ingredients or excipients may affect the mechanical properties of the material; as a consequence, they may adversely affect the process or the product.
• Changes in the packaging material, e.g. replacing plastics by glass, may require changes in the packaging procedure and therefore affect product stability.
• Changes in the process, e.g. changes in mixing time, drying temperature and cooling regime, may affect subsequent process steps and product quality.
• Changes in equipment, including measuring instruments, may affect both the process and the product; repair and maintenance work, such as the replacement of major equipment components, may affect the process.
• Changes in the production area and support system, e.g. the rearrangement of manufacturing areas and/or support systems, may result in changes in the process. The repair and maintenance of support systems, such as ventilation, may change the environmental conditions and, as a consequence, revalidation/requalification may be necessary, mainly in the manufacture of sterile products.
• Unexpected changes and deviations may be observed during self-inspection or audit, or during the continuous trend analysis of process data.
Periodic revalidation. It is well known that process changes may occur gradually even if experienced operators work correctly according to established methods. Similarly, equipment wear may also cause gradual changes. Consequently, revalidation at scheduled times is advisable even if no changes have been deliberately made.
The decision to introduce periodic revalidation should be based essentially on a review of historical data, i.e. data generated during in-process and finished product testing after the latest validation, aimed at verifying that the process is under control. During the review of such historical data, any trend in the data collected should be evaluated.
In some processes, such as sterilization, additional process testing is required to complement the historical data. The degree of testing required will be apparent from the original validation.
Additionally, the following points should be checked at the time of a scheduled revalidation:
• Have any changes in master formula and methods, batch size, etc., occurred? If so, has their impact on the product been assessed?
• Have calibrations been made in accordance with the established programme and time schedule?
• Has preventive maintenance been performed in accordance with the programme and time schedule?
• Have the standard operating procedures (SOPs) been properly updated?
• Have the SOPs been implemented?
• Have the cleaning and hygiene programmes been carried out?
• Have any changes been made in the analytical control methods?
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