We reported previously that the methanolic root extract of the Indian medicinal plant Pluchea indica Less.(Asteraceae) could neutralize viper venom-induced action [Alam, M.I., Auddy, B., Gomes, A., 1996. Viper venom neutralization by Indian medicinal plant (Hemidesmus indicus and P. indica) root extracts. Phytother. Res. 10, 58-61]. The present study reports the neutralization of viper and cobra venom by beta-sitosterol and stigmasterol isolated from the root extract of P. indica Less.(Asteraceae). The active fraction (containing the major compound beta-sitosterol and the minor compound stigmasterol) was isolated and purified by silica gel column chromatography and the structure was determined using spectroscopic analysis (EIMS,(1)H NMR,(13)C NMR). Anti-snake venom activity was studied in experimental animals. The active fraction was found to significantly neutralize viper venom-induced lethal, hemorrhagic, defibrinogenation, edema and PLA(2) activity. Cobra venom-induced lethality, cardiotoxicity, neurotoxicity, respiratory changes and PLA(2) activity were also antagonized by the active component. It potentiated commercial snake venom antiserum action against venom-induced lethality in male albino mice. The active fraction could antagonize venom-induced changes in lipid peroxidation and superoxide dismutase activity. This study suggests that beta-sitosterol and stigmasterol may play an important role, along with antiserum, in neutralizing snake venom-induced actions.

The present study reports the isolation and purification of lupeol acetate from the methanolic root extract of Indian medicinal plant Hemidesmus indicus (L.) R.Br.(family: Asclepiadaceae) which could neutralize venom induced action of Daboia russellii and Naja kaouthia on experimental animals. Lupeol acetate could significantly neutralize lethality, haemorrhage, defibrinogenation, edema, PLA(2) activity induced by Daboia russellii venom. It also neutralized Naja kaouthia venom induced lethality, cardiotoxicity, neurotoxicity and respiratory changes in experimental animals. Lupeol acetate potentiated the protection by snake venom antiserum action against Daboia russellii venom induced lethality in male albino mice. Venom induced changes in lipid peroxidation and super oxide dismutase activity was antagonized by lupeol acetate. Snake venom neutralization by lupeol acetate and its possible mechanism of action has been discussed.

Pathophysiology due to snakebite is a combined effect of various actions of the complex venom constituents. Importance of protein toxins in snake envenomation is well known. The present investigation reports the existence of nonprotein/nonpetide low molecular weight toxin in Indian King Cobra venom, which plays an important role in envenomation consequences in experimental animal models. A group of non-peptidic toxins (OH-NPT1) was isolated from Indian King Cobra Ophiophagus hannah by thin layer chromatography and silica gel column chromatography. UV, IR, NMR and (ESI) TOF-MS studies characterized the OH-NPT1 as a mixture of aliphatic acids having molecular weights 256, 326 and 340Da. The minimum lethal dose of OH-NPT1 was found to be 2.5 microg/20g (iv) and 4microg/20g (ip) in male albino mice. The cardiotoxic property of OH-NPT1 was established through studies on isolated guinea pig heart and auricle preparations, ECG studies in albino rat and estimation of LDH1/LDH and CPK-MB/CPK ratio in Swiss albino mice. Commercial antiserum failed to neutralize the lethality and cardiotoxicity of the toxin. However, calcium and magnesium effectively neutralized the lethal action.

In the present study, King Cobra (Ophiophagus hannah) venom was subjected to TLC followed by column chromatography/HPLC to isolate and purify a non-protein toxin designated as KC-MMTx.(1)H NMR, IR and EIMS studies showed KC-MMTx likely to be a 282 D unsaturated aliphatic acid having molecular formula C(18)H(34)O(2). The minimum lethal dose of KC-MMTx was 200mug/kg (i.v.) and 350mug/kg (i.p.) in Swiss albino male mice. It significantly increased pentobarbitone induced sleeping time and significantly decreased the body temperature of male albino mice. It provided protection against amphetamine aggregate toxicity in mice but failed to protect amphetamine stereotypy in male albino rats. KC-MMTx provided significant protection against drug (strychnine, pentylenetetrazole, yohimbine) induced convulsions in male albino mice. It increased serum Na(+) and decreased serum Ca(2+) significantly in male mice. MAO activity and brain neurotransmitter levels in male mice were altered significantly. Further detailed study is warranted on the CNS, anticonvulsant potential of KC-MMTx, which may lead to the development of newer therapeutic tools in the near future.

In recent years, mesenchymal stem cells (MSC) have been attracting the greatest interest in the regeneration of injured tissues, autoimmune diseases, and transplantation of hematopoietic progenitor cells. Bone marrow (BM) represents the major source of MSC; however, umbilical cord blood (UCB) MSC has some advantages over BM, such as the higher differentiation capability and noninvasive collection methods. We sought to establish a 7-color, single-tube flow cytometric assay to quantify MSC in fresh tissues, namely BM and UCB, based on phenotypic markers of these cells. Moreover, we evaluated the differential expression of these markers in BM and UCB MSC. We used 5 UCB samples and 5 BM samples obtained from individuals without hematologic disease. To characterize MSC we used the following combination of monoclonal antibodies: CD71-FITC; CD105-PE; CD184-PE-Cy5; CD34-PE-Cy7; CD133-APC; CD45-APC-H7; CD44-Pacific blue, acquiring at least 1 million nucleated cells. We observed a greater number of BM MSC when compared with UCB MSC as well as some differences in the expression of some MSC antigens, particularly CD105 and CD44. Based on our preliminary results, phenotypic identification of MSC by flow cytometry is possible using a 7-color, single-tube assay. However, culture assays after sorting of cells characterized in this study are required to prove that they correspond to MSC.

Considering that short bouts of stretching, as recommended in rehabilitation and sports activities, induce skeletal muscle and connective tissue adaptation, the hypothesis of this study was that MMP-2 activity is regulated by muscle stretch. The level of MMP-2 activity was, thereby, assessed after stretching in rat soleus muscle. Animals received a single session of stretching (10 stretches lasting 1 min each with 30 s of rest in between) and were evaluated immediately and after 8, 24, 48, 72 and 168 h. To evaluate the effect of repetitive sessions of stretching, three groups of animals were evaluated - one group after 2 sessions, another after 3, and a third after 7. MMP-2 activity was evaluated by zymography and MMP-2 mRNA was assessed by real-time polymerase chain reaction. None of the groups presented MMP-9 activity. MMP-2 activity and mRNA expression did not change after either single or repetitive sessions of stretching. In conclusion, the results of this study indicate that MMP-2 is not involved in the muscle stretch-induced remodeling.

The growing incidence of microbial infections and the increasing ability of such organisms to acquire resistance to antimicrobial treatment lead the requirement of fast bacteria and fungi identification methods. In this work we explored optical spectroscopic techniques on fungal identification. We show that some fungal infections can be identified by ultraviolet optical excitation of fungi fluorescence followed by the spectral analysis of the emitted light. Moreover, we demonstrate that ultraviolet LED and LASER could be applied in fungal identification and a new device for fungal diagnosis is proposed.

BACKGROUND: The potential relationships between sleep-wake behaviors and emotional/disruptive problems in otherwise healthy school-aged children are unclear. METHODS: A parental questionnaire was developed for the epidemiologic survey of children's sleep and wake behavioral patterns. The questions covered a wide range of features including sleep length (school days, weekends), time to fall asleep, night awakenings, bedtime and nighttime sleep-related behaviors, daytime sleepiness, irritability, and tiredness. To assess psychiatric symptomatology, the Rutter Scale B2 was completed by teachers. In addition to the total score, sub-scores of emotional, hyperactivity, and conduct problems were obtained. The representative population sample comprised 779 children (403 girls), with an age range of 6-11 years. RESULTS: Hyperactivity and conduct problems at school in boys were both associated with parental reports of bedtime resistance. Hyperactivity was also associated with longer sleep duration during weekends. Conduct and emotional problems in girls were associated with earlier bedtime during school days. Emotional problems in girls were also associated with longer sleep durations in school days and weekends. CONCLUSION: Bedtime resistance was the only sleep behavior associated with either hyperactivity or conduct problems in children, and longer sleep durations appear to occur more frequently in children with both hyperactive or emotional problems. Information about good sleep hygiene at bedtime may help parents setting sleep limits.

Smoking is an important cause of pulmonary pathology and this addiction can be regarded as a chronic, recurrent disea- se. The benefits of smoking cessation are unquestionable and all physicians should become more active and assertive in recommending it. Aim: to characterize population that seek medical support for smoking cessation and understand why some are suc- cessful and others are not. Material and methods: retrospective analysis of medi- cal records of outpatients in follow-up between January 2003 and June 2006. Age, gender, age of smoking initia- tion, smoking burden (number of packs-years), associa- ted diseases, degree of dependence (Fargerstrom test for nicotine dependence), previous attempts of and motiva- tion for smoking cessation, therapy, rate of smoking ces- sation, need for behavioural support and abandonment were evaluated. Results: five hundred and twenty six patients studied, 50% male with an average age of 45,5 +/- 11,4 years. Al- most half (43,1 %- n=227) of the patients started smoking before the age of 15. Average smoking burden was 35,8 +/- 20 pack-years although 21,4%(n=113) smoked more than 50 pack-years. Respiratory disease was present in 52,1%(COPD- 39,9% and others - 12,2%) and car- diovascular disease in 14,6% of the patients. In 46% of patients (n=242), a relevant psychiatric disorder was iden- tified; depression (21,4%), anxiety disorder (19,4%), other dependencies (2,1%) bipolar disorder (1,5%) and schizo- phrenia (0,6%). The evaluation of degree of dependence revealed maximum level in 69,7% of the patients (n=380). Many patients (72,2%- n=380) referred earlier quitting attempts. The strongest motivation factors for giving up smoking were concern for health (83,5%), financial issues (8,2%) and search for better quality of life (5,7%). Most patients (81,7%- n=430) were submitted to nicotine re- placement therapy; transdermal patches (53,3%), chewing gum (1,1%) or both (45,6%). Psycopharmacological tre- atment included ansiolytic therapy (86,5%), bupropion hydrochloride (2,3%) and antidepressant (0,6%). Seventy six patients (14,4%) benefited from behavioural support. Two hundred and twenty three patients (42,4%) were af- ter one year successful while 219 (41,6%) abandoned fo- llow up, most after the first time. Most patients that aban- doned follow up referred lack of motivation and price of therapy. Conclusions: The population in study had a high rate of psychiatric disorders, high level of dependence and lack of motivation which might justify the rate of abandonment. Success in therapy was associated with close follow-up, beha- vioural support and pharmacological therapy. Key-words: Smoking cessation, outpatient smokers.

Snakebite envenomations cause severe local tissue necrosis and the venom metalloproteinases are thought to be the key toxins involved. In this study, the ethanolic extract from seed kernels of Thai mango (Mangifera indica L. cv.'Fahlun')(Anacardiaceae) and its major phenolic principle (pentagalloylglucopyranose) exhibited potent and dose-dependent inhibitory effects on the caseinolytic and fibrinogenolytic activities of Malayan pit viper and Thai cobra venoms in in vitro tests. molecular docking studies revealed that the binding orientations of the phenolic principles were in the binding pockets of snake venom metalloproteinases (SVMPs). The phenolic principles could form hydrogen bonds with the three histidine residues in the conserved zinc-binding motif and could chelate the Zn(2+) atom of the SVMPs, which could potentially result in inhibition of the venom enzymatic activities and thereby inhibit tissue necrosis.

Seed of Mucuna pruriens (Velvet beans) has been prescribed by traditional medicine practitioners in Nigeria as a prophylactic oral antisnake remedy. In the present studies, we investigated the protective effects of M. pruriens seed extract (MPE) against histopathological changes induced by intravenous injection of Naja sputatrix (Malayan cobra) venom in rats pretreated with the seed extract. Examination by light microscope revealed that the venom induced histopathological changes in heart and blood vessels in liver, but no effect on brain, lung, kidney and spleen. The induced changes were prevented by pretreatment of the rats with MPE. Our results suggest that MPE pretreatment protects rat heart and liver blood vessels against cobra venom-induced damages.

Although anti-venom therapy is available for the treatment of fatal bite by snakes, it offers less or no protection against the local effects such as dermo- and myonecrosis, edema, hemorrhage and inflammation at the bitten region. The viper species are known for their violent local effects and such effects have been commonly treated with plant extracts without any scientific validation in rural India. In this investigation, the methanolic extract of grapes (Vitis vinifera L.) seed was studied against the Indian Daboia/Vipera russelli venom-induced local effects. The extract abolished the proteolytic and hyaluronidase activities and also efficiently neutralized the hemorrhage, edema-inducing and myonecrotic properties of the venom. In addition, the extract also inhibited partially the pro-coagulant activity of the venom and abolished the degradation of Aalpha and Bbeta chains of human fibrinogen. Thus, the extract possesses potent anti-snake venom property, especially against the local effects of viper bites.

ETHNOPHARMACOLOGICAL RELEVANCE: The seed, leaf and root of Mucuna pruriens have been used in traditional medicine for treatments of various diseases. In Nigeria, the seed is used as oral prophylactics for snakebite. AIM OF THE STUDY: To study the protective effects of Mucuna pruriens seed extract against the lethalities of various snake venoms. MATERIALS AND METHODS: Rats were pre-treated with Mucuna pruriens seed extract and challenged with various snake venoms. The effectiveness of anti-Mucuna pruriens (anti-MPE) antibody to neutralize the lethalities of snake venoms was investigated by in vitro neutralization. RESULTS: In rats, MPE pre-treatment conferred effective protection against lethality of Naja sputatrix venom and moderate protection against Calloselasma rhodostoma venom. Indirect ELISA and immunoblotting studies showed that there were extensive cross-reactions between anti-MPE IgG and venoms from many different genera of poisonous snakes, suggesting the involvement of immunological neutralization in the protective effect of MPE pre-treatment against snake venom poisoning. In vitro neutralization experiments showed that the anti-MPE antibodies effectively neutralized the lethalities of Asiatic cobra (Naja) venoms, but were not very effective against other venoms tested. CONCLUSIONS: The anti-MPE antibodies could be used in the antiserum therapy of Asiatic cobra (Naja) bites.

The ethanolic extract from seed kernels of Thai mango (MSKE)(Mangifera indica L. cv.'Fahlun')(Anacardiaceae) and its major phenolic principle (pentagalloyl glucopyranose) exhibited dose-dependent inhibitory effects on enzymatic activities of phospholipase A(2)(PLA(2)), hyaluronidase and L-amino acid oxidase (LAAO) of Calloselasma rhodostoma (CR) and Naja naja kaouthia (NK)venoms by in vitro tests. The anti-hemorrhagic and anti-dermonecrotic activities of MSKE against both venoms were clearly supported by in vivo tests. Molecular docking studies indicated that the phenolic molecules of the MSKE could selectively bind to the active sites or their proximity, or modify conserved residues that are critical for the catalysis of PLA(2), and selectively bind to the LAAO binding pocket of both CR and NK venoms and thereby inhibit their enzymatic activities. The results imply a potential use of MSKE against snake venoms.

Ethnobotanical surveys were conducted in four different indigenous groups in Southern parts of Tamilnadu, India, using a questionnaire. The herbal practitioners in the study area were interviewed, and information on medicinal plants was collected from the traditional healers called "Vaidyars". This survey covers 72 medicinal plants belonging to 53 families that are used for the treatment of snakebite in a traditional way. Traditional approach was evaluated scientifically with some selected plant extracts (7.2mg/kgbw) and partially purified fractions (2.4mg/kgbw) were orally administered to mice experimentally envenomed with rattlesnake venom s.c. injection (2.5-15mug/kgbw). Tested fractions (Aristolochia indica, Hemidesmus indicus, Gloriosa superba, Strychnos nux-vomica, Eclipta prostrata, and Andrographis paniculata) showed potent neutralizing effect against the venom. Compared to the extracts, administration of purified fractions was more effective in increasing the body weight. Control mice injected with the venom alone showed weight loss and severe toxicity at 15mug/kgbw. The purified fractions (2.4mg/kgbw) produced significant protection against venom induced changes in serum SOD and LPx levels. The isolated fractions effectively inhibited the toxic effect of snake venoms in vitro than in vivo. The above observations confirmed the protective activity of plants-Aristolochia indica, Hemidesmus indicus, Gloriosa superba, Strychnos nux-vomica, Eclipta prostrata, and Andrographis paniculata against the lethal action of snake venom and need further investigation.