Extraction, purification and analysis of conotoxin of Conus textile captured from Persian Gulf and the investigation of analgesic effects of conotoxins in an animal model

Identification of venomous species of Persian Gulf cone snails and characterization of venom composition and their features is so important from the point of medical importance. Marine cone snails from the genus Conus are estimated to consist of up to 700 species. The venom of cone snails has yielded a rich source of novel neuroactive peptides or conotoxins. The present study was aimed to study the analgesic effect of Persian Gulf Conus textile and its comparison with morphine in mouse model. The specimens of Conus textile were collected of Larak Island from depth of 7 m. The collected samples were transferred to laboratory alive and were stored at -700 c. he veno s ducts were separated and ho ogenized with deionized water he ixture centrifuged at rp for inutes upernatant was considered as extracted veno and stored at - C after lyophylization. The protein profile of venom determined by using SDS-PAGE and HPLC used to investigate the extracted venom and to evaluate the analgesic activity, formalin test was carried out. SDS-PAGE indicated several bands ranged between 6 and 250 kDa. Chromatogram of the venom demonstrated more than 44 large and small fractions. The amount of 10 ng of Conus crude venom and analgesic peptide showed the best anti-pain activity in formalin test. No death observed up to 100 mg/kg, which is 250,000 times higher than the effective dose.Venom characterization of Persian Gulf Conus textile may be of medical importance and potential for new pharmaceutical drugs as well.

Venomics and biological analysis of Scatophagus argus argus (Spotted scat) venom isolated in Persian Gulf, extraction and purification of hemolytic protein

Green scat namely as Scatophagus argus argus is a venomous aquarium fish belonging to Scatophagidae family. It can induce painful wounds in injured hand with partial paralysis to whom that touch the spines. Dorsal and ventral rough spines contain cells that produce venom with toxic activities. According to unpublished data collected from local hospitals in southern coastal region of Iran, S. argus is reported as a venomous fish. Envenomation induces clinical symptoms such as local pain, partial paralysis, erythema and itching. In the present study green scat (spotted scat) was collected from Persian Gulf coastal waters. SDS-PAGE indicated 12 distinct bands in the venom ranged between 10-250 KDa. The crude venom had hemolytic activity on human erythrocytes (1%) with an LC100 (Lytic Concentration) of about 1.7 μg. The crude venom can release 813 μg proteins from 0.5% casein. Phospholipase C activity was recorded at 3.125 μg of total venom. Our findings showed that the edematic activity remained over 48 h after injection. The purification of the venom was done by HPLC and 30 peaks were obtained within 80 min but only one peak in 68 min retention time showed hemolytic activity at 90% acetonitril was isolated. The area percentage of the hemolytic protein showed that this hemolytic protein consist of 32 percent of total proteins and its molecular weight was 72 KDa in SDS_PAGE. The results demonstrated that crude venom extracted from Iranian coastal border has different toxic and enzymatic activities.

Report of the sub-regional workshop to validate the scoping study of nutrient pollution on the coastal and marine systems of South Asia, Colombo, Sri Lanka, 21-22 May, 2014

The workshop agenda included: presentations from collaborative institutions, national governments and resource persons; a draft scoping study on nutrient loading; and an ecosystem approach to pollution management was tested.

Understanding nutrient loading and sources in the Bay of Bengal Large Marine Ecosystem

Inputs of nitrogen, phosphorous and dissolved silica from watersheds draining into the Bay of Bengal Large Marine Ecosystem are calculated for the present day and predictions made for 2030 and 2050 are presented. The major sources are identified and the Indicator of Coastal Eutrophication (ICEP) is calculated.

Liver cytochrome oxidase P4501A1 purification of Acipenser persicus and designing ELISA for measuring

Moon oxygenases related to cytochrome P450s are the molecular Biomarkers which have important role in Biotransformation of endogenous and exogenous compounds and catalazyin of many biological reactions. One of the important isoenzyme is cytochrome P4501A. This isoenzyme involved in metabolism of environment pollutnts such as PAHs. Because of its inducibility, it has a key tool for impact assesment of contaminants in aquatic environment. In this study, at first, that fractions containing Acipenser persicus and Huso huso isoenzyme were purified, and after that Antibodies against them were prepared. For isolation of isoenzyme fraction, Microsomes were prepared from fish liver using differential centrifugation at high speeds. microsomes were solubiized by cholat sodium and Emulgen. Extraction of this isoenzyme was done with the combinatuion of ionexchange chromatography and gelfiltration or chromatofocusing chromatography. Ion exchange chromatography and gel filtration were applied in DEAE sepharose fast flow and sephacryl S200 respectively and chromatofocusing was done at poly buffer 74 and 94 exchanger. The results of SDS-PAGE Showed that the molecular weight of isoenzyme was about 58±1 KDa. Furthermore the inmunoblotting results confirmed this subject. Isoenzyme activigy based on EROD (Ethoxyresorofin o-deethylase) reaction showed about 20-26 fold increase in enzyme activity of treated fish than control fish. The results of Elisa, Using monoclonal anti cod P4501A demonstrated the inducibility and highly elevated of its activity in treated sample more than the control fish. Mean while, the fish sample were showed the strong reaction to polyclonal antibody against beluga P4501A1 prepared in our Lab compared to monoclonal anti body.

Nutrient status and thermal stratification in Lake Victoria

Worldwide, human activity in the watershed has been found to induce lake responses at various levels, including at population and ecosystem scale. Recently, Carignan and Steedman (2000) reported on disruptions of biogeochemical cycles in temperate lakes following watershed deforestation and lor wildfire and Carignan et al., (2000 a, b) concluded that water quality and aquatic biota are strongly influenced by disturbances in the watershed. Similarly, Lake Victoria is no exception as people in its catchment have exploited it for the last hundred years or more, but have now begun to understand the extent to which they have thrown the lake into disorder and how their increasing activity in the watershed have driven some environmental changes within and around the lake.

Responses to seasonal changes in nutrient quality and patchiness of food in a multigroup community of Tibetan macaques at Mt. Emei

Provisioning along pedestrian trails by tourists much increased the nutrient quality and patchiness of food (NqPF)for Tibetan macaques (Macaca thibetana) at Mt Emei in spring and summer. In the habitat at a temperate-subtropical transition zone, the mncaque's NqPF could be ordered in a decreasing rank from spring summer to autumn to winter With the aid of a radio-tracking system, I collected ranging data on a multigroup community in three 70-day periods representing the different seasons in 1991-92, Rank-order correlation on the data show that with the decline of NqPF; the groups tended to increase days away from the trail, their effective range size (ERS) their exclusive area (EA) and the number of days spent in the EA, and reduced their group/community density and the ratio of the overlapped range to the seasonal range (ROR). In icy/snowy winter; the macaques searched for mature leaves slowly and carefully in the largest seasonal range with a considerable portion that was nor used in other seasons. Of the responses, the ROR decreased with the reduction in group/community density; and the ERS was the function of both group size (+) and intergroup rank (-) when favorite food was highly clumped. All above responses were clearly bound to maximize foraging effectiveness and minimize energy expenditure, and their integration in term of changes in time and space leads to better understanding macaque ecological adaptability. Based on this study and previous work on behavioral and physiological factors, I suggest a unifying theory of intergroup interactions. Ir! addition, as the rate of behavioral interactions,was also related to the group density, I Waser's (1976) gas model probably applies to behavioral, as well as spatial, data on intergroup interactions.

Purification, characterization and primary structure of a chymotrypsin inhibitor from Naja atra venom

A chymotrypsin inhibitor, designated NA-CI, was isolated from the venom of the Chinese cobra Naja atra by three-step chromatography. It inhibited bovine (x-chymotrypsin with a K-i of 25 nM. The molecular mass of NA-CI was determined to be 6403.8 Da by matrix-assisted laser-desorption ionization time-of-flight (MALDI-TOF) analysis. The complete amino acid sequence was determined after digestion of S-carboxymethylated inhibitor with Staphylococcus aureus V8 protease and porcine trypsin. NA-CI was a single polypeptide chain composed of 57 amino acid residues. The main contact site with the protease (PI) has a Phe, showing the specificity of the inhibitor. NA-CI shared great similarity with the chymotrypsin inhibitor from Naja naja venom (identities = 89.5%) and other snake venom protease inhibitors. (C) 2003 Elsevier Inc. All rights reserved.

Separation and purification of ovulation-inducing factors in the seminal plasma of the Bactrian camel (Camelus bactrianus)

Ovulation in the Bactrian camel depends upon ovulation-inducing factors in the seminal plasma. The present study was conducted to isolate and purify the bioactive fractions from the seminal plasma of these camels. The seminal plasma was fractionated by anion-exchange chromatography, and six fractions were obtained. The bioactive potential of each fraction was estimated from its effect on rat pituitary tissue cultured in vitro and by the effect of an intramuscular injection of the fraction into female camels in vivo. Both the third fraction (F3) and the fifth fraction (F5) stimulated the release of LH in vitro and in vivo. In addition, female camels ovulated within 48 h after intramuscular injection of F3. However, neither F3 nor F5 had any significant effect on the secretion of FSH, either in vitro or in vivo. When F3 was further fractionated into four subfractions, the third subfraction (F3-3) still stimulated the in vitro release of LH, but not of FSH. An attempt to further purify the ovulation-inducing factors in F3-3 failed owing to the similarity of the molecular characters.

Purification, characterization and biological activities of the L-amino acid oxidase from Bungarus fasciatus snake venom

L-Amino acid oxidases (LAAOs) are widely distributed in snake venoms, which contribute to the toxicity of venoms. However, LAAO from Bungarus fasciatus (B. fasciatus) snake venom has not been isolated previously. In the present study, LAAO from B. fasciat

Expression, purification and characterization of recombinant Jerdonitin, a P-II class snake venom metalloproteinase comprising metalloproteinase and disintegrin domains

Jerdonitin is a P-II class snake venom metalloproteinase comprising metalloproteinase and disintegrin domains. In this study, we established a high-level expression system in Pichia pastoris and developed a purification strategy for the recombinant Jerdonitin. This recombinant Jerdonitin degraded fibrinogen at a level of activity comparable with its wild type. The effects of recombinant Jerdonitin on inhibiting ADP-induced human platelet aggregation were in a dose-dependent manner with an IC50 of 248 nM. In addition, we reported here that Jerdonitin can significantly inhibit the growth of several cell lines, including human liver cancer cells (Bel7402), human leukemia cells (K562) and human gastric carcinoma cells (BGC823). This study offers recombinant Jerdonitin that will be valuable for further functional and structural studies of Jerdonitin. (C) 2009 Elsevier Ltd. All rights reserved.

Ventilation effectiveness measures based on heat removal: Part 2. Application to natural ventilation flows

The three effectiveness measures based on the ability of a flow to flush buoyancy from a ventilated space proposed by Coffey and Hunt [Ventilation effectiveness measures based on heat removal-part 1. Definitions. Building and Environment, in press, doi:10.1016/j.buildenv.2006.03.016.] are applied to assess and compare two fundamental natural ventilation flows. We focus on the limiting cases of passive displacement and passive mixing ventilation flows during transient conditions. These transient flows occur when, for example, heat is purged from a building at night. Whilst it is widely recognised that mixing flows are less efficient at purging heat than displacement flows, our results indicate that, when a particular zone of a room is considered, displacement ventilation can result in lower effectiveness than mixing ventilation. When a room is considered as a whole, displacement ventilation yields higher effectiveness than mixing ventilation and we quantify these differences in terms of the geometry of the space and opening area. The proposed theoretical predictions are compared with effectiveness deduced from measurements made during laboratory experiments and show good agreement. © 2006 Elsevier Ltd. All rights reserved.

Ventilation effectiveness measures based on heat removal: Part 1. Definitions

The effectiveness of ventilation flows is considered from the perspective of buoyancy (or heat) removal from a space. This perspective is distinct from the standard in which the effectiveness is based on the concentrations of a neutrally buoyant contaminant/passive tracer. Three new measures of effectiveness are proposed based on the ability of a flow to flush buoyancy from a ventilated space. These measures provide estimates of instantaneous and time-averaged effectiveness for the entire space, and local effectiveness at any height of interest. From a generalisation of the latter, a vertical profile of effectiveness is defined. These measures enable quantitative comparisons to be made between different flows and they are applicable when there is a difference in density (as is typical due to temperature differences) between the interior environment and the replacement air. Applications, therefore, include natural ventilation, hybrid ventilation and a range of forced ventilation flows. Finally, we demonstrate how the ventilation effectiveness of a room may be assessed from simple traces of temperature versus time. © 2006 Elsevier Ltd. All rights reserved.