Analgesic and anti-inflammatory activities of Piper nigrum L.

ObjectiveTo evaluate and compare the analgesic and anti-inflammatory activity of pure compound, piperine along with hexane and ethanol extracts of Piper nigrum L. fruit in mice and rats.MethodsThe analgesic activity was determined by tail immersion method, analgesy-meter, hot plate and acetic acid induced writhing test. While the anti-inflammatory activity was evaluated by carrageenan-induced paw inflammation in rats.ResultsPiperine at a dose of 5 mg/kg and ethanol extract at a dose of 15 mg/kg after 120 min and hexane extract at a dose of 10 mg/kg after 60 min exhibited significant (P<0.05) analgesic activity by tail immersion method, in comparison to ethanol extract at a dose of 10 mg/kg using analgesy-meter in rats. However, with hotplate method, piperine produced significant (P<0.05) analgesic activity at lower doses (5 and 10 mg/kg) after 120 min. A similar analgesic activity was noted with hexane extract at 15 mg/kg. However, in writhing test, ethanol extract significantly (P<0.05) stopped the number of writhes at a dose of 15 mg/kg, while piperine at a dose of 10 mg/kg completely terminated the writhes in mice. In the evaluation of anti-inflammatory effect using plethysmometer, piperine at doses of 10 and 15 mg/kg started producing anti-inflammatory effect after 30 min, which lasted till 60 min, whereas hexane and ethanol extracts also produced a similar activity at a slightly low dose (10 mg/kg) but lasted for 120 min.ConclusionsIt is concluded from the present study that Piper nigrum L possesses potent analgesic and anti-inflammatory activities.     

Phytochemical and pharmacological evaluation of prop roots of Pandanus fascicularis Lam

ObjectiveTo evaluate the anti-inflammatory and analgesic activities of the ethanol and aqueous extracts of prop roots of Pandanus fascicularis (P. fascicularis) Lam (pandanaceae). And provide experimental evidence for its traditional use such as rheumatoid arthritis and spasmodic.MethodsThe anti-inflammatory activity was observed by carrageenan-induced edema of the hind paw of rats. Analgesic activities of prop roots of P. fascicularis were determined using acetic acid induced writhing model and tail clip method in mice and rat, respectively. The ethanol fraction was then subjected to chromatographic analysis and a compound has been isolated and characterized by IR, ¹H-NMR and mass spectroscopy.ResultsEdema suppressant effect of ethanol extract was found to be 37.03% inhibition whereas aqueous extract was found to be 63.22% inhibition after 3 h which was nearly equivalent to that of 10 mg/kg of indomethacin (67.81%). Percentage inhibition of writhing compared to control were 63.15%, 54.38%, 14.90% for aspirin, aqueous extract and ethanolic extract, respectively. Both ethanol and aqueous extracts show significant activity against appropriate controls after 60 min of treatment on tail clip method. The structure of the isolated compound is may be characterized as Hepta deca-5-ene-1-ol by analysis it's IR, ¹H-NMR and mass spectroscopy data.ConclusionsThe extracts of prop roots of P. fascicularis produce significant analgesic and anti-inflammatory activities, supporting the traditional application of this herb in treating various diseases associated with inflammation and pain.     

Pentacyclic triterpenoic acids: new chemoprotective compounds. Minireview

Ursolic acid and oleanolic acid are pentacyclic triterpenoic acids having a similar chemical structure and are the major components of some oriental and traditional medicine herbs wildly distributed all over the world. There is a growing interest in the elucidation of the biological roles of both these triterpenoid compounds. This review summarizes the biological activities of presented triterpenoid acids (anti-inflammatory, hepatoprotective, gastroprotective, anti-ulcer, anti-HIV, cardiovascular, hypolipidemic, antiatherosclerotic and immunoregulatory effects). Our interest has been focussed especially on their anti-tumor and chemopreventive activity. Both compounds have been shown to act at various stages of tumor development, including inhibition of tumorigenesis, inhibition of tumor promotion, and induction of tumor cell differentiation. They effectively inhibit angiogenesis, invasion of tumor cells and metastasis. However, the mechanisms by which they act are poorly understood. Ursolic acid and oleanolic acid are relatively non-toxic and could be use as chemopreventive/chemoprotective agents in clinical praxis.     

Pharmacological properties and related constituents of stem bark of Pterocarpus erinaceus Poir. (Fabaceae)

ObjectiveTo screen methanol and dichloromethane extracts of stem bark of Pterocarpus erinaceus for anti-inflammatory, analgesic, in vitro antioxidant activities and phytochemical analysis.MethodsAnti-inflammatory activity was determined by using carrageenan induced-edema of mice paw and croton oil-induced edema of mice ear; analgesic effect was evaluated using acetic acid-induced writhing. Phytochemical screening of extracts was performed by thin layer chromatography. The chromatographic fractionation led to the isolation of main active components as friedelin, lupeol and epicathechin. The structures were established by TLC and nuclear magnetic resonance studies.ResultsBoth methanol and dichloromethane extracts, friedelin, lupeol and epicatechin showed a significant anti-inflammatory effect using croton oil induced-ear edema. Furthermore, the action of dichloromethane extract was more important. At the doses of 100 and 200 mg/kg, the methanol extract was able to reduce the carrageenan induced-hind paw edema, while at the doses of 100, 200 and 400 mg/kg, it showed an important analgesic effect against writhing induced by acetic acid injection of 38.8%, 68.0% and 74.3%, respectively. Antioxidative properties of methanol extract and its dichloromethane and ethyl acetate fractions were assessed by using the 1,1-diphenyl-2-picrylhydrazyl method. The methanol extract showed the stronger radical scavenging activity than dichloromethane and ethyl acetate fractions, with an antiradical power of 5, 3.5 and 2 respectively. The main components isolated from these extracts as friedelin, lupeol and epicathechin were responsible of these activities.ConclusionsThe results suggest that the stem bark extracts of Pterocarpus erinaceus possessed important anti-inflammatory, analgesic activities and strong antioxidant properties, therefore, they could be used as natural potential ingredients for pharma ceutical industry.     

Analgesic and anti-inflammatory activities of Passiflora foetida L

ObjectiveTo investigate the analgesic and anti-inflammatory activities of ethanol extract of Passiflora foetida (P. foetida) leaves.MethodsEthanol extract of P. foetida leaf was evaluated for analgesic action by acetic acid-induced writhing and hot plate method in albino mice. The anti-inflammatory property of ethanolic leaf extract was tested by carrageenan induced acute paw edema and histamine induced acute paw edema in rats.ResultsThe dose 200 mg/kg of P. foetida leaf extract exhibited highest significant analgesic activity [(13.50±0.43) min] at a reaction time of 20 min in hot plate method in mice. The ethanol extract of leaf dose 100 mg/kg produced a highly significant anti inflammatory effect [(1.302±0.079) mL] in rats.ConclusionsIt is very clear that P. foetida also has analgesic and anti-inflammatory activities for the pharmaceuticals.     

Development of Large-Scale Fractionation Methods

Abstract. A method is described for large-scale isolation of prothrombin complex from Cohn fraction III by means of polyethylene glycol (PEG 4000) and DEAE-cellulose. Residual thrombin and active factor X were irreversibly inactivated in the prothrombin complex preparation by the addition of heparin and cofactor, which in turn was isolated from Cohn fraction IV by means of Al(OH)₃ and ammonium sulfate.
By this method a prothrombin complex preparation was obtained which was concentrated 29-fold over plasma in factor IX activity, with a 59-fold purification. The other three clotting factors were concentrated 17–51-fold with 36–106-fold purification. These concentrates met all the DBS requirements for human factor IX complex.
The presence of activated factor IX and its inactivation during the stabilization step are postulated from the activity diagrams obtained by preparative acrylamide disc electrophoresis of prothrombin complex before and after addition of cofactor and heparin.
Partial separation of antithrombin III and heparin cofactor activities was observed following fractionation of our cofactor by preparative disc electrophoresis.
Isoelectric focusing failed to separate individual clotting factors of the prothrombin complex.     

Melatonin protects against gastric ulceration and increases the efficacy of ranitidine and omeprazole in reducing gastric damage

The antiulcer effect of melatonin on gastric lesions caused by restraint-cold stress was studied with the intent of determining the mechanism of action of this agent. Melatonin dose-dependently prevented restraint-cold stress-induced gastric damage with around 90% inhibition at a dose of 60 mg/kg BW. When compared with already marketed antiulcer drugs such as ranitidine and omeprazole, melatonin was found to be more effective than ranitidine but less effective than omeprazole in preventing stress ulcer. As stress-induced gastric lesions are mainly caused by oxidative damage because of hydroxyl radicals (*OH), the effect of melatonin in scavenging the.OH generated during stress conditions in vivo as well as in an in vitro model system were studied. The results indicate that melatonin caused an 88% reduction of endogenous *OH during stress in vivo, an observation confirmed in an established in vitro system. Furthermore, a decrease in the activity of gastric peroxidase (GPO) and an increase in the gastric mitochondrial superoxide dismutase (Mn-SOD) activity because of restraint-cold stress was attenuated by melatonin pretreatment indicating that the indole possibly exerts its gastroprotective effects through its direct as well as indirect antioxidant activities. Moreover, in separate experiments, cotreatment of rats with melatonin and ranitidine or omeprazole was found to protect against stress ulceration in doses at which either of these alone could not protect the stomach. The findings raise the possibility of melatonin being considered as an effective gastroprotective agent individually or as a cotreatment with either ranitidine and omeprazole.     

Evaluation of in vitro antioxidant and apoptotic activities of Cyperus rotundus

Objective:To evaluate in vitro antioxidant and apoptotic activities of Crperus rotundas(C.rotundus).Methods:The phytochemical study and the antioxidant activities of both methanol and aqueous extracts from C.rotundus aerial part were determined.In addition,these extracts were also investigated for their cytotoxic and apoptotic activities.The major compound of the methanol extract was isolated.Both metlianol and aqueous extracts(300,150,and 50μg/mL)were evaluated for their antioxidant activity by the xanthine/xanthine oxidase assay system.However,16,8,and 4 mg/mL of each extract were tested to investigate their OH.formation scavenging potential.Aqueous extract(800,400.and 200μg/mL)and melhunol extract(350,175,and 88μg/mL)were tested against lipid peroxidation,induced by 75μM H_2O_2,The cytotoxicity(by MTT assay)and cell DNA fragmentation of both extracts were evaluated Inwards K562 and L1210 cell lines.The major compound was obtained from the butanol fraction of methanol extract and its structure was determined by KMN spectroscopic analysis.Results:The methanol and aqueous extracts showed respectively,88%and 19%inhibition of xanthine oxidase activity.Vet.the same extracts inhibited lipid peroxidation by 61.5%and 42.0%.respectively.Roth extracts inhibited OH.formation by 27.1%and 25.3%,respectively.Only methanol cxtract induced DNA degradation.Orientin was determined as the major compound isolated from the butanol fraction of metlianol extract.Conclusions:It appears that C.rotundus extracts exhibit a potential use as a natural antioxidant and an apoptosis inducer.     

Antinociceptive and anti-inflammatory activities of crude extracts of Ipomoea involucrata leaves in mice and rats

ObjectiveTo investigate antinociceptive and anti-inflammatory activities of crude extract from Ipomoea involucrata leaves (Convolvulaceae) in mice and rats.MethodsThe antinociceptive activity was tested using acetic acid-induced abdominal writhing test in mice. The anti-inflammatory activity was evaluated using egg albumin induced oedema of rat paw.ResultsPhytochemical screening showed the presence of alkaloids, flavonoids, saponins, terpenoids and tannin. At the doses of 25–100 mg/kg, Ipomoea involucrata exhibited dose-dependent and significant increase in pain threshold in acetic acid –induced writhing test of mice (P<0.05, student t-test) The administration of Ipomoea involucrata leaf extract (25–100 mg/kg) showed dose-dependent decreases in paw volume of egg albumin induced oedema in rats and a significant higher anti-inflammatory activity compared to the standard control (Aspirin).ConclusionsThese results support the claims on the traditional use of the of Ipomoea involucrata leaves in the treatment of toothache, rheumatic pains and other inflammatory conditions. Studies on the isolation and structural elucidation of the active principle are still needed being carried out.     

Evaluation of anti-inflammatory property of the leaves of Sansevieria liberica ger. and labr. (fam: Dracaenaceae)

ObjectiveTo evaluate the anti-inflammatory property of leaves of Sansevieria liberica Ger. and Labr. and to ascertain the toxicity and phytochemical profiles of the extract of the leaves.MethodsThe juice from the fresh leaves was expressed manually and lyophilized. The crude extract (CE) was then fractionated into n-hexane fraction (HF), chloroform fraction (CF), ethylacetate fraction (EF) and methanol fraction (MF). The crude extract (CF) and the fractions were screened for anti-inflammatory activity using egg albumen-induced paw (systemic) edema in rats as a measure of acute inflammation. The toxicity test and phytochemical screening were done using standard procedures.ResultsThe CE and the fractions significantly (P<0.05) inhibited the development of paw edema induced by egg albumen in rats. The potency/activity of the CE and the fractions increased in the order HF>CE>MF>CF>EF, with the CE and HF at 400 mg/kg exhibiting inhibition comparable to that obtained with 5 mg/kg diclofenac sodium. Acute toxicity test on CE established an oral and intraperitoneal LD₅₀ of > 5 000 mg/kg in mice. Phytochemical screening of the CE and the fractions showed the presence of various bioactive substances such as alkaloids, saponins, flavonoids, terpenoids, steroids, glycosides, reducing sugars, tannins, resins, carbohydrates, proteins, acidic compounds, fats and oils.ConclusionsThe results of the study showed that the leaves of Sansevieria liberica Ger and Labr. possess anti-inflammatory effects which may be due to its bioactive constituents. Further purification on these bioactive constituents may result in the development of potent anti-inflammatory agent with low toxicity and better therapeutic index.     

Aspirin esterase activity - Evidence for skewed distribution in healthy volunteers

BACKGROUND: Aspirin, with its analgesic, anti-inflammatory, antipyretic, and anti-platelet actions, is one of the most frequently used drugs. Although its use as prophylaxis against thromboembolism is well established, an optimal dose, conferring maximal anti-platelet action without increased risk of bleeding, remains elusive. METHOD: We assessed the possible pharmacokinetic contribution to this problem in 107 healthy, non-medicated volunteers. Serum aspirin esterase activity was evaluated at 37 degrees C with 1 mM aspirin as substrate. On the basis of the report that most of aspirin esterase activity is accounted for by pseudocholinesterase, we additionally quantified the activity of this enzyme, with and without dibucaine as an inhibitor, using Ellman's reaction, in 41 of our volunteers. RESULTS: Aspirin esterase activities in all of our volunteers (33.90 nmol/ml/min to 222.65 nmol/ml/min, median 103.45 nmol/ml/min) showed a continuous and skewed distribution with eight outliers. In the 41 subjects so studied, aspirin esterase activities correlated positively with both pseudocholinesterase activities (Spearman's rho=0.593, p<0.001) and dibucaine numbers (Spearman's rho=0.422, p<0.01). CONCLUSIONS: Our results support previous observations that the rate of aspirin hydrolysis is not determined by aspirin esterase alone and that other factors are probably involved. Additionally, the skewed distribution of aspirin esterase activities makes a case for its possible contribution to the phenomenon of aspirin resistance.     

Gastroprotection by an aluminium- and magnesium hydroxide-containing antacid in rats. Role of endogenous prostanoids

This study was designed to determine the gastroprotective actions of an antacid (Maalox 70) and its components, Al(OH)3 and Mg(OH)2, against acute gastric lesions induced by absolute ethanol, acidified aspirin (ASA), and water immersion and restraint stress in rats. Given orally, the antacid prevented dose-dependently the formation of gastric lesions by all three ulcerogens, and these effects were similar to those obtained with a methylated prostaglandin E2 (PGE2) analog. Active Al(OH)3 gel was equipotent with Maalox, whereas Mg(OH)2 was significantly less effective in gastroprotection than Maalox 70. Chemically inactive Al(OH)3 wet gel showed only small and insignificant protective properties. Since the gastroprotective activities of Maalox 70 against ethanol lesions cannot be reversed by pretreatment with indomethacin, and since neither Maalox 70 nor its active components affected the mucosal generation of PGE2 and leukotriene C4, we postulate that mucosal prostanoids are not the primary mediators in the mechanism of their protective action on the gastric mucosa.     

Analgesic and anti-inflammatory effects of honey: the involvement of autonomic receptors

The use of honey for therapeutic purposes is on the increase and many studies have shown that honey has the ability to influence biological systems including pain transmission. Therefore, this study was designed to investigate the analgesic and anti-inflammatory effects of honey and the effects of concurrent administration of autonomic nervous system blocking drugs. Studies on analgesic activities was carried out using hotplate and formalin-induced paw licking models while the anti-inflammatory activity was by the carrageenan paw oedema method. Animals were distributed into six groups consisting of five animals each. They were administered saline, honey (600 mg/kg), indomethacin (5 mg/kg), autonomic blockers (3 μg/kg of tamsulosin, 20 mg/kg (intraperitoneally) of propranolol, 2 ml/kg of atropine or 10 mg/kg (intra muscularly) of hexamethonium) or honey (200 and 600 mg/kg) with one of the blockers. The results showed that honey reduced pain perception especially inflammatory pain and the administration of tamsulosin and propranolol spared the effect of honey. Hexamethonium also spared the effects of honey at the early and late phases of the test while atropine only inhibited the early phase of the test. However, atropine and hexamethonium spared the anti-inflammatory effects of honey but tamsulosin abolished the effects while propranolol only abolished the anti-inflammatory effects at the peak of the inflammation. The results suggest the involvement of autonomic receptors in the anti-nociceptive and anti-inflammatory effects of honey although the level of involvement depends on the different types of the receptors.     

Melatonin protects against piroxicam-induced gastric ulceration

The antiulcer effect of melatonin on gastric lesions caused by piroxicam was studied with the intent of determining the mechanism of action of this agent. Melatonin dose-dependently lowered piroxicam and indomethacin-induced gastric damage with more than 90% inhibition at a dose of 60 mg/kg BW. Increased lipid peroxidation, augmented protein oxidation and decreased glutathione content of the gastric tissue following piroxicam treatment indicated a possible involvement of oxidative stress in this nonsteroidal anti-inflammatory drug (NSAID)-induced gastropathy. Pretreatment of rats with melatonin prevented these changes. Oral administration of piroxicam to rats caused a threefold increase in the tissue levels of hydroxyl radical generation, a change significantly attenuated by melatonin. Furthermore, a decrease in the activity of gastric peroxidase and an increase in the activity of gastric superoxide dismutase(s) (SOD) because of piroxicam treatment was attenuated by melatonin pretreatment indicating that the indole possibly exerts its gastroprotective effects through its direct as well as indirect antioxidant activities. The results of the present studies also reveal that melatonin may influence the expression of Cu-Zn SOD, catalase, cyclooxygenase as well as alpha-actinin whose levels were found to be altered, following piroxicam treatment. The current studies, therefore, document melatonin's gastroprotective ability against piroxicam-induced gastric damage and the findings raise the possibility of melatonin being considered as a co-therapy with piroxicam or other NSAIDs in reducing the gastropathy when long-term use of these nonsteroidal agents are unavoidable.     

Antidiabetic principles of Loranthus micranthus Linn. parasitic on Persea americana

ObjectiveTo explore antidiabetic principles of the Eastern Nigeria mistletoe, Loranthus micranthus Linn. parasitic on Persea Americana.MethodsThe weakly acidic fraction of the aqueous methanol extract of the leaves of Loranthus micranthus (Linn.) was isolated and tested for its antidiabetic activities. The isolation of the weakly acidic fraction was carried out following established physico-chemical based procedures. Furthermore, alloxan-induced diabetic rats were treated intraperitoneally (ip) with 250 mg/kg and 400 mg/kg of the weakly acidic fraction, glibenclamide 10 mg/kg (positive control) and 2 mg/kg of 3 % v/v tween 20 (negative control). The sugar levels of the treated and untreated animals were determined by withdrawing the blood at regular intervals and testing them with an automated glucometer. The phytochemical analysis of the acidic fraction was carried out using standard procedures. Chromatographic techniques were employed in the subsequent isolation and purification of the constituents of the weakly acidic fraction.ResultsIt was shown that the maximum effect of the weakly acidic fraction was obtained at 24 hours after administration and was found to be statistically comparable with that of the positive control. The phytochemical analysis revealed the presence of steroids, terpenoids, alkaloids, flavonoids, glycosides, carbohydrates, saponins, and acidic compounds in the crude extract and carbohydrates, flavonoids, terpenoids and oil in the weakly acidic fraction. Further purification of the weakly acidic fraction of the methanol extract using thin layer chromatography shows that toluene: methanol: diethyl amine (3:1:1) and chloroform: methanol: diethyl amine (9:1:1) are the best solvent system for the isolation of the various components of the weakly acidic fraction of the crude methanol extract of Loranthus micranthus.ConclusionsThe present study has led to the conclusion that the weakly acidic fraction of the plant under study has the potent antidiabetic activity and that the various components can be isolated using basic chromatographic techniques.     

Phytochemical,antioxidant and hepatoprotective effects of different fractions of Moringa oleifera leaves methanol extract against liver injury in animal model

Objective: To evaluate the potential antioxidant and hepatoprotective effects of n-hexane, dichloromethane(DCM), ethyl acetate(EtOAc), n-butanol and aqueous fractions of Moringa oleifera(M. oleifera) leaves methanol extract against carbon tetrachloride(CCl_4)-induced liver injury in rats. Methods: These fractions were prepared from the M. oleifera leaves methanol extract by solubilization in water and partitioning in n-hexane, EtOAc, DCM and n-butanol. Their phyto-components were identified by GC-MS analysis. The in vitro antioxidant effect of these fractions was carried out by assessment of 1,1-diphenyl-2-picrylhydrazyl scavenging activity. A total of 40 Sprague Dawley rats were allocated into 8 equal groups: group 1 given olive oil(1 m L/kg b.wt.), group 2 injected with CCl_4, group 3 to 7 administered with n-hexane, DCM, EtOAc, n-butanol and aqueous fractions, respectively after CCl_4, group 8 administered with silymarin after CCl_4. The activities of aspartate aminotransferase, alanine aminotransferase, and the levels of total cholesterol, triglycerides, glucose, total proteins and albumin in serum were determined spectrophotometrically. Glutathione reduced, lipid peroxide by-products levels, glutathione-s-transferase and catalase enzyme activities in the liver homogenate were determined by spectrophotometer. Liver specimens were also examined for histopathological alterations under light microscope. Results: The GCMS analysis of different fractions of the M. oleifera leaves methanol extract revealed that n-hexane, DCM, EtOAc, n-butanol, and aqueous fractions contained 17, 22, 23, 19 and 32 compounds, respectively. The percent and the molecular structure of each component in each fraction were identified. The n-butanol and EtOAc fractions exhibited the strongest in vitro antioxidant activity against 1,1-diphenyl-2-picrylhydrazyl. CCl_4 significantly decreased glutathione reduced and total proteins concentration and glutathione-s-transferase and catalase activities but increased lipid peroxide by-products and total cholesterol levels. The n-hexane followed by aqueous and DCM fractions were the most potent to regulate serum enzyme activities and lipid peroxide by-products levels in the liver homogenate. Conclusions: n-hexane, DCM, and aqueous fractions have the highest effectiveness against CCl_4-induced hepatotoxicity. Isolation and purification of the active constituents require further experiments.     

Characteristics of the Factor VIII Protein and Factor XIII in Various Factor VIII Concentrates

The in vitro properties of 5 factor VIII preparations (AHF-Kabi, Hemofil Hyland, AHF-Profilate Abbott, Kryobulin Immuno and Factorate High Purity Armour) and an ordinary cryoprecipitate were studied with reference to factor VIII clotting activity (VIII:C), factor VIII clotting antigen (VIILCAg), factor VIII related antigen (VIIIR:Ag) (EI, IRMA, CIE), ristocetin cofactor activity (VIIIR:RCF), fibrinogen and factor XIII. All the preparations with the exception of Factorate had higher levels of VIII:CAg than VIII:C indicating inactivation of the biological activity of VIII:C during the procedure. AHF-Kabi (fraction I-0) and the cryoprecipitate, the only preparations capable of normalising the defect in patients with von Willebrand's disease, showed the same level of VIIIR:Ag determined by EI and by IRMA, while all the other preparations (i.e. cryoprecipitates purified further in different ways) had considerably lower levels of VIIIR:Ag determined by IRMA than by EL Based on these in vitro techniques it seems to be possible to predict which preparations can be used successfully in patients with von Willebrand's disease, while no such conclusions can be made from VIIIR:RCF determinations. EI yielded similar concentrations of factor XIII a subunit in all the preparations tested. 3 functional assays showed high factor XIII activities in AHF-Kabi but low or no activities in the others. Thus, considerable differences were found of the in vitro properties of the proteins in 5 factor VIII concentrates and a cryoprecipitate. The action of proteases and the techniques used in the purification procedure are probably of crucial importance for the properties of the various factors.     

The effect of purification procedures on the determination of serum level of 1,25-dihydroxyvitamin D

The advance in knowledge of clinical disorders involving calcium and bone diseases has resulted in part from the development of assay procedures for the major vitamin D metabolites, but the values of the same samples measured in different laboratories vary considerably. They suggest that the chromatographic purification involved in these assays seems to be a critical step. In this study, we compared the results of 1,25-dihydroxyvitamin D (1,25(OH)2D) using different chromatographic purification methods: Sephadex LH-20 column chromatographic purification (non-HPLC method) described by Mallon et al. and High Performance Liquid Chromatographic purification (HPLC method) modified by Yamaoka et al. Serum 1,25(OH)2D levels of healthy volunteers before and after oral load of 4 micrograms of 1 alpha-hydroxyvitamin D3 (1 alpha OHD3) were measured by the two different methods mentioned above. A good correlation (r = 0.951, p less than 0.001) was noted in the samples before loading. The mean values of serum 1,25(OH)2D before loading did not differ significantly between HPLC method and non-HPLC method. After loading, however, the mean value of serum 1,25(OH)2D was significantly higher in non-HPLC method (p less than 0.01). The values of 1,25(OH)2D were determined in the serum added 1,25-dihydroxy-24-oxo-vitamin D3 (24-oxo-D3) (0 pg/ml, 200 pg/ml or 2000 pg/ml). In the samples added 2000 pg/ml of 24-oxo-D3, the mean value of 1,25(OH)2D was significantly higher in non-HPLC method (p less than 0.001). The affinity of 24-oxo-D3 for 1,25(OH)2D3 cytosolic receptor was evaluated in chick embryo intestinal mucosa and rachitic chick intestinal mucosa.(ABSTRACT TRUNCATED AT 250 WORDS)     

Antipyretic, anti-inflammatory and analgesic properties of nilavembu kudineer choornam: a classical preparation used in the treatment of chikungunya fever

ObjectiveTo investigate the efficacy of ethanolic extract of nilavembu kudineer choornam (EENKC) in inflammation, pain and fever using animal models to support its actions.MethodsAcute toxicity study of EENKC was performed in mice to fix the effective dose. The antipyretic, anti-inflammatory and analgesic activity of EENKC was evaluated in brewer's yeast induced pyrexia in rats, carrageenan-induced inflammation in rats and acetic-acid induced writhing in mice model.ResultsAcute toxicity revealed that EENKC didn't show death and toxic signs up to 2 000 mg/kg. In brewer's yeast induced pyrexia and carrageenan-induced inflammation EENKC at the doses of 200 and 400 mg/kg inhibited fever and inflammation significantly (P<0.01 and <0.05) compared to control animals. In mice, the number of writhing induced by acetic-acid was significantly (P<0.01) reduced after treatment with both the dose of EENKC than control animals. EENKC 200 mg/kg inhibits inflammation higher level in carrageenan-induced paw edema, but there is no significant difference when compared to indomethacin 10 mg/kg.ConclusionsThe present findings revealed that EENKC possesses antipyretic, anti-inflammatory and analgesic activity which supports nilavembu kudineer choornam efficacy in chikungunya fever.     

Cathepsin B: association with plasma membrane in metastatic tumors.

The subcellular localization of cathepsin B activity (EC 3.4.22.1) in three murine melanomas of increasing metastatic potential (Cloudman less than B16-F1 less than B16 amelanotic) was determined. Cathepsin B activity was localized in the heavy mitochondrial fraction of normal murine liver but in the light mitochondrial fraction of the metastatic melanomas; the localization of three other lysosomal hydrolases did not shift. Further purification of the light mitochondrial fraction into L-1 (density = 1.045 g/ml) and L-2 (density = 1.07 g/ml) fractions was achieved on a 30% iso-osmotic Percoll gradient. The L-1 fraction of liver and melanomas contained Na+, K+-ATPase activity; the L-2 fraction of liver contained four lysosomal hydrolase (cathepsins B and H, N-acetyl-beta-glucosaminidase, and beta-glucuronidase) and glucose-6-phosphatase activities. Ultrastructural examination revealed that the L-1 fraction consisted of membrane vesicles and the L-2 fraction of secondary lysosomes. In the B16 melanomas cathepsin B and N-acetyl-beta-glucosaminidase activities were found in both L-1 and L-2 fractions. Specific activities of the two enzymes in the plasma membrane (L-1) fractions increased in correspondence with metastatic potential. Cathepsin H and beta-glucuronidase activities were not localized in the plasma membrane fractions of the B16 melanomas. Localization of hydrolytic enzymes in the plasma membrane of metastatic tumor cells could result in focal dissolution of the extracellular matrix and thereby invasion and metastasis.