Cobra Research Articles

JENIS SATWA LIAR YANG DIPERDAGANGKAN SEBAGAI BAHAN PANGAN DI KOTA PONTIANAK

This research aims to analyze animal species which are in trade as livestock. In purpose of investigating the conservation status of animals, it is to review the animal species. Method of this research was descriptive with qualitative approach. Data analysis to be applied was survey. The samples of analysis were the ministerial regulation on environmental and forestry number 92 of 2018, CITES, and Constitution number 5 of 1990. Result of the research showed that there were nine sorts of animal that were in trade as livestock. The best sold sorts of animal are cobra snakes, monitor lizards, and squirrels. The supplies were from Ketapang, Pontianak, Purun, Rasau. From Kalimantan Tengah, the supplies were specialized in wild boars and cobra snakes. Based on the price, wild boars and turtles had been the most expensive which are Rp. 70.000/kg. Four variants of foods in menu included sauced, soy sauced, rica, and fried. There were five species of animals which are conserved based on the ministerial regulation on environmental and forestry number 92 of 2018 such as monitor lizards and turtles and the other species such as cobra snakes, bats, and squirrels which are on appendix CITES II.Keywords : Ethnozoology, Livestock, Wild Animals.

Changes and clinical significance of serum secretory phospholipase A2 in cobra bite patients

Objective To investigate the relationship between serum secreted phospholipase A2 (sPLA2) and the severity of the disease in cobra bite patients. Methods Fifty-two cobra bite patients admitted to our hospital from January 2015 to December 2015 were selected according to the inclusion and exclusion criteria. According to the degree of disease, patients were divided into the light and heavy risk groups. According to the degree of swelling of the affected limbs, patients were divided into the mild swelling group and moderate-severe swelling group. According to the time difference between visits, patients were divided into: visit time <8 h group and visit time≥ 8 h group. Twenty healthy adult volunteers served as the control group. The levels of serum sPLA2, C reactive protein (CRP), D-dimer (DD), lactic acid (LAC) and peripheral blood leukocyte count (WBC) were measured and compared in different groups. Quantitative data were analyzed using t test and rank sum test. Results The levels of serum sPLA2 in the light and heavy risk group were statistically different from those in the control group (P<0.05), and were statistically significant between the light and heavy risk groups (P<0.01) and between the mild swelling group and moderate-severe swelling group (P<0.05). The serum sPLA2 levels in group A and group B were also significantly different from those in the control group (P<0.05). The level of sPLA2 was positively correlated with the level of LAC, DD and WBC, and the correlation coefficients were 0.314 2, 0.275 2 and 0.653 4, respectively. Conclusions The higher the level of serum sPLA2, the more serious the patient's poisoning symptoms. It is of certain clinical value to evaluate the condition of cobra. Key words: Secretory phospholipase A2; Cobra bite; Lactic acid; D-dimer; C-reactive protein

In vitro activity of isolated fractions from Iranian cobra snake venom against Plasmodium falciparum

Background: Malaria is one of the most important parasitic diseases worldwide with high morbidity and mortality in tropical and subtropical areas. Available drugs for the treatment of Malaria are expensive, have side effects and/or there may be some drug-resistant species. Resistance to most antimalarial drugs has encouraged the development of new drugs. An replacement source for discovering such drugs is natural products. Venom is a complex mixture of active pharmaceutical ingredients. The purpose of this study was investigation of antimalarial activity of the fractions obtained from the venom of Iranian cobra, Naja naja oxiana. Methods & Materials: Lyophilized Persian Kobra snake venom of was obtained from the Toxicology department of Pasteur Institute of Iran. It was purified by gel filtration from a Sephacryl S-200 HR column. It was then followed by FPLC on a Mono Q ion exchange column. After the first column chromatography the fractions were lyophilized. Based on results five fractions were collected. The concentration of fractions determined and their anti-malaria activity estimated in vitro and the effective fraction was purified by the second column. SDS PAGE was carried out to test for purity of the effective fraction. Plasmodium falciparum 3D7 were grown in vitro by the method of Trager and Jensen. When the parasites reached 10%, the fractions were tested by the checker board method for 50% inhibitory concentration of the parasites as checked by counting method using giemsa stain. Results: The Third fraction did not have hemolytic effect at different concentrations. One of the venom fractions (0.02 mg/ml) with half maximal inhibitory concentration (IC50) value of 0/002 mg/ml after 48 hr with significance of p < 0.001was seen to be the most effective fraction against plasmodium falciparum (ring stage). The Third fraction was purified and was seen to be 63 KDa on SDS-PAGE. It was observed that this fraction reduced the viability of parasite and strongly inhibited of growth it. Conclusion: The Active fraction with higher efficacy and lower toxicity of Naja naja oxiana had an inhibitory effect on Plasmodium falciparum. The further studies are recommended to evaluate in vivo effects in animal models.

Application of modified negative pressure closed drainage technique in early treatment of cobra bite ulcer

Objective To investigate the application of modified negative pressure closure drainage in early stage of snakebite. Methods 60 patients with early bite ulcer were selected as study objects, and they were randomly divided into two groups according to the digital table.30 patients in the observation group received vacuum sealing drainage(VSD) combined with chymotrypsin continuous rinsing treatment.30 patients in the control group were given conventional dressing bandaging treatment.The two stage skin grafting time, wound healing time, complications, disability rate, hospitalization time and C reactive protein(CRP) levels of the two groups were observed.And the treatment effect was analyzed. Results The incidence rates of complications and disability in the observation group were 0.00% and 3.33%, respectively, which were significantly lower than 16.67% and 26.67% in the control group, and the differences between the two groups were statistically significant(χ2=4.356, 6.405, all P<0.05). The CRP levels of 3 d and 7 d in the observation group were (15.8±1.7)mg/L, (6.9±1.6)mg/L, respectively, which in the control group were (18.6±2.3)mg/L, (13.8±1.9)mg/L, respectively, there were statistically significant differences between two groups(t=5.362, 15.214, all P<0.01). The two stage skin grafting, wound healing, hospital stay of the observation group were (9.5±1.8)d, (24±1.8)d, (25.3±1.9)d, respectively, which were significantly shorter than those of the control group [(16.2±2.6)d, (32.0±3.2)d, (33.2±3.7)d], the differences were statistically significant(t=11.604, 11.934 10.403, all P<0.01). Conclusion For patients with injury of cobra bite to early ulcer, using VSD technology combined with chymotrypsin continuous irrigation, can significantly reduce the level of CRP and the occurrence of complications and disability rate, shorten the time of two stage skin grafting, wound healing time, hospitalization time, reduce the pain of patients, it is the best treatment to accelerate recovery. Key words: Snake bites; Ulcers; Drainage; Chymotrypsin

Analysis of correlation between RASSF2A gene methylation and colorectal cancer

Objective To investigate the methylation status of RASSF2A gene promoter in colorectal cancer tissues and its correlation with the occurrence of colorectal. Methods From February 2007 to March 2013, a total of 111 cases of sporadic colorectal cancer patients were selected. The methylation status of RASSF2A gene in tumor tissue and normal mucosa were detected by using combined bisulfite restriction endonuclease method (COBRA). The correlations between the methylation of RASSF2A gene and the clinicopathologic features of colorectal cancer were analyzed. Results Methylation of RASSF2A gene was found in 83.78% (93/111) and 7.21% (8/111) of colorectal cancer tissues and normal tissues, with statistically significant difference (χ2=131.245, P<0.001). The methylation of RASFF2A gene was not correlated with histology type (χ2=3.554, P=0.314), Duke staging (χ2=3.217, P=0.359) and location of tumor (χ2=1.060, P=0.303). Conclusion The presence of RASSF2A hypermethylation in colorectal cancer may be associated with the occurrence of colorectal cancer. Key words: Colorectal neoplasms; DNA methylation; RASSF2A gene

Novel Naja atra cardiotoxin 1 (CTX-1) derived antimicrobial peptides with broad spectrum activity.

Naja atra subsp. atra cardiotoxin 1 (CTX-1), produced by Chinese cobra snakes, belonging to Elapidae family, is included in the three-finger toxin family and exerts high cytotoxicity and antimicrobial activity too. Using as template mainly the tip and the subsequent β-strand of the first “finger” of this toxin, different sequences of 20 amino acids linear peptides have been designed in order to avoid toxic effects but to maintain or even strengthen the partial antimicrobial activity already seen for the complete toxin. As a result, the sequence NCP-0 (Naja Cardiotoxin Peptide-0) was designed as ancestor and subsequently 4 other variant sequences of NCP-0 were developed. These synthesized variant sequences have shown microbicidal activity towards a panel of reference and field strains of Gram-positive and Gram-negative bacteria. The sequence named NCP-3, and its variants NCP-3a and NCP-3b, have shown the best antimicrobial activity, together with low cytotoxicity against eukaryotic cells and low hemolytic activity. Bactericidal activity has been demonstrated by minimum bactericidal concentration (MBC) assay at values below 10 μg/ml for most of the tested bacterial strains. This potent antimicrobial activity was confirmed even for unicellular fungi Candida albicans, Candida glabrata and Malassezia pachydermatis (MBC 50–6.3 μg/ml), and against the fast-growing mycobacteria Mycobacterium smegmatis and Mycobacterium fortuitum. Moreover, NCP-3 has shown virucidal activity on Bovine Herpesvirus 1 (BoHV1) belonging to Herpesviridae family. The bactericidal activity is maintained even in a high salt concentration medium (125 and 250 mM NaCl) and phosphate buffer with 20% Mueller Hinton (MH) medium against E. coli, methicillin resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa reference strains. Considering these in vitro obtained data, the search for active sequences within proteins presenting an intrinsic microbicidal activity could provide a new way for discovering a large number of novel and promising antimicrobial peptides families.

Dynamic changes of serum protein in rats with acute intoxication of Chinese cobra snake venom by proteomic analysis

To elucidate the toxic mechanism of snake venom at the protein level, proteomics technology was applied to investigate the effect of venom on circulation in the mammalian body. Temporal proteomic analysis was performed to profile the dynamic changes in the sera of Sprague–Dawley rats administered with Chinese cobra venom or saline. Using 8-plex iTRAQ analysis, 392 and 636 serum proteins were identified to be linearly upregulated or downregulated over time in the low-dose group and high-dose group, respectively. These proteins were mainly associated with the acute phase response pathway, complement system, and liver X receptor (LXR)/retinoid X receptor (RXR) and farnesoid X receptor (FXR)/RXR activation pathways. Compared with the low-dose group, the immune response and integrin pathways were inhibited in the high-dose group, although no obvious effect was observed. With consistently higher or lower expression in the high-dose group compared to the low-dose group throughout the whole process of venom poisoning, two proteins, Kininogen-1 (KNG1) and orosomucoid 1 (ORM1), which are involved in metabolism and immune response, occupied a core position in the pathway network and are considered venom dose-dependent biomarker candidates.

Study on the correlation between Ras related sequence family 2A gene methylation and colorectal cancer in feces

Objective To investigate the value of methylation of Ras related sequence family 2A (RASSF2A) gene methylation in the stool of patients with colorectal cancer in the clinical diagnosis of RASSF2A. Methods A modified COBRA method was used to analyze the methylation status of 102 RASSF2A gene promoter regions (region 1 and region 2) in two cases of colorectal cancer. 51 cases of healthy subjects as control group negative colonoscopy. At the same time, the relationship between the methylation of RASSF2A gene and clinical pathological factors. Results In the stools of patients with colorectal cancer extensive methylation, methylation and methyl of were 87% and 63% (64/102) 89/102 and 13% (13/102) and 6% (3/51) and 22% (11/51) and 78% (40/51) in the control group respectively. The detection rate of methylation in stool of colorectal cancer patients was significantly higher than that of healthy persons, and the rate of non methylation was significantly lower than that of the control group, the difference was statistically significant (P=0.000). There was no significant correlation between the methylation of RASSF2 gene and the gender, tumor stage, and the degree of invasion of colorectal cancer patients. Conclusion Methylation of RASSF2A gene is an early event in the progression of colorectal cancer. Methylation of RASSF2 gene in feces is expected to be a new approach to the early diagnosis of colorectal cancer. Key words: Ras related sequence family 2A gene; Methylation; Colorectal cancer

Effect of cobra venom factor on the immune function of the pig to monkey liver xeno-transplantation

Objective To explore the effect of high purity venom factor (CVF) and other factors on immune function after pig to monkey liver xeno-transplantation. Methods To establish the pig to monkey heterotopic auxiliary liver transplantation model, the occurrence of super acute rejection (HAR) and delayed rejection (DXR) was observed after liver transplantation while interfered by high purity CVF. The expression of IgG, IgM and C3 protein in liver tissues was detected by Western blotting at different time points. Results The donor liver used for the adsorption all occured HAR, respectively of 10, 12, 11, 9, 13 minutes, the average time was (11.00±1.41) minutes. The pathological findings were ischemic necrosis and thrombosis. After CVF and other multiple factors, all the recipient monkey liver did not observe HAR and DXR until death, the IgG, IgM and C3 protein deposition in liver tissue after the death of the receptor were lower than the donor liver used for the adsorption, which is statistically significant difference (P=0.026, 0.047, 0.032). Conclusion CVF and other multiple factors can effectively overcome the occurrence of HAR in pig to monkey liver xeno-transplantation, and has the potential to prevent or delay the DXR. Key words: Xenograft; Snake venom factor; Rejection

Impedimetric Biosensor for the Determination of Phospholipase A2 Activity in Snake Venom

ABSTRACTAn impedimetric biosensor to determine the activity of phospholipase A2 has been developed for the first time. This biosensor is based on the immobilization of enzyme phospholipase A2 (PLA2) from snake venom on the surface of a gold electrode. The enzymatic reaction takes place in the presence of L-α-phosphatidylcholine (lecithin) substrate, in 10 mM pH 7.4 phosphate buffer. The impedance of enzymatically modified electrode/electrolyte interface increased as the concentration of lecithin increases. A detection limit of 10−11 M and a linear range of 10−10 to 10−5 M were obtained. At a fixed concentration of lecithin (10−3 mM), when the PLA2 enzyme with a certain level of activity is injected, the impedance of the enzymatically modified electrode/electrolyte interface changed in the inverse direction, due to the competitive effect of injected PLA2 enzyme. A linear range of 10 to 400 ng/L (corresponding to 16 to 640 U/L) was obtained for commercial naja mossambica mossambica snake venom. The activity of PLA2 from naja cobra snake venom (Origin Thailand) was determined to be 75% of that of naja mossambica mossambica snake venom.

Relationship between hippocampal CREB/BDNF signaling pathway and cognitive dysfunction in rats with chronic pathological pain

Objective To evaluate the relationship between hippocampal cyclic adenosine monophosphate response element-binding protein(CREB)/brain-derived neurotrophic factor(BDNF)signaling pathway and cognitive dysfunction in rats with chronic pathological pain. Methods Thirty-two healthy adult male Sprague-Dawley rats, weighing 220-250 g, were divided into 3 groups using a random number table: control group(group C, n=8), sham operation group(group S, n=8)and chronic pathological pain group(group CP, n=16). Chronic pathological pain model was established by injecting cobra venom 0.4 mg(4 μl)into the sheath of the infraorbital nerve.The mechanical pain threshold was measured at 3 days before establishment of the model(baseline)and 4 days and 1, 2, 3, 4 and 8 weeks after establishment of the model.Morris water maze test was performed to evaluate the spatial learning and memory abilities at 5 and 9 weeks after establishment of the model.Eight rats were sacrificed at 5 and 9 weeks after establishment of the model in CP group, and rats were sacrificed after the end of Morris water maze test at 9 weeks after establishment of the model in C and S groups.The hippocampi were isolated for determination of the expression of phosphorylated CREB and BDNF in the hippocampal tissues using Western blot. Results Compared with group C, the mechanical pain threshold was significantly decreased at each time point after establishment of the model, the escape latency was prolonged at 5 and 9 weeks after establishment of the model, the rate of time of staying at the target quadrant was decreased, the frequency of crossing the original platform was decreased, and the expression of phosphorylated CREB and BDNF was down-regulated at 9 weeks after establishment of the model in group CP(P 0.05). Conclusion The mechanism underlying cognitive dysfunction may be related to inhibited activation of CREB/BDNF signaling pathway in the hippocampus of rats with chronic pathological pain. Key words: Pain; Cognition disorders; Cyclic AMP response element-binding protein; Brain-derived neurotrophic factor

Hannah Arendt and the Art of Secrecy; Or, the Fog of Cobra Mist

Hannah Arendt and the Art of Secrecy; Or, the Fog of Cobra Mist

Analgesic effect of curcumin on trigeminal neuralgia in rats

Objective To evaluate the analgesic effect of curcumin on trigeminal neuralgia in rats. Methods Thirty healthy adult male Sprague-Dawley rats, weighing 200-250 g, aged 7-8 weeks, were divided into 3 groups (n=10 each) using a random number table: sham operation group (group S), trigeminal neuralgia group (group TN) and trigeminal neuralgia + curcumin group (group Cur). Trigeminal neuralgia model was established by injecting cobra venom solution into the sheath of the infraorbital nerve (ION). Starting from 15 days after establishment of the model, curcumin 45 mg/kg was intragastrically administered twice a day for 28 consecutive days in group Cur, while the equal volume of peanut oil was administered in group TN.Before establishment of the model (baseline), and on 4, 7, 14, 21, 28, 35 and 42 days after establishment of the model, the mechanical pain threshold was measured, the free behaviors were observed, and the time and frequency of face-grooming and exploratory behaviors were recorded.After observation of the free behaviors, the ION and medulla oblongata on the affected side were removed for examination of the ultrastructure using transmission electron microscopy. Results Compared with group S, the mechanical pain threshold was significantly decreased, the time of face-grooming behaviors was significantly prolonged, the frequency of face-grooming behaviors was significantly increased, the time of exploratory behaviors was significantly shortened, and the frequency of exploratory behaviors was significantly decreased on 4-42 days after establishment of the model in group TN (P<0.05 or 0.01). Compared with group TN, the mechanical pain threshold was significantly increased, and the time of face-grooming behaviors was significantly shortened on 28-42 days after establishment of the model, and the frequency of face-grooming behaviors was significantly decreased, the time of exploratory behaviors was significantly prolonged, and the frequency of exploratory behaviors was significantly increased on 21-42 days after establishment of the model in group Cur (P<0.05 or 0.01). Microscopic examination revealed that the changes in demyelination of the ION and medulla oblongata were significantly attenuated in group Cur as compared with group TN. Conclusion Intragastrically administered curcumin 45 mg/kg (twice a day for 28 consecutive days) can attenuate trigeminal neuralgia in rats, and the mechanism is related to the attenuated changes in demyelination of the ION and medulla oblongata. Key words: Curcumin; Trigeminal neuralgia; Analgesic

Effect of curcumin on cognitive dysfunction in a rat model of trigeminal neuralgia

Objective To evaluate the effect of curcumin on cognitive dysfunction in a rat model of trigeminal neuralgia. Methods Thirty healthy adult male Sprague-Dawley rats, weighing 200-250 g, were randomly divided into 3 groups (n=10 each) using a random number table: sham operation group (Sham group), trigeminal neuralgia group (TN group), and trigeminal neuralgia+ curcumin group (TN+ Cur group). Trigeminal neuralgia was produced by injecting cobra venom solution 4 μl into the sheath of the infraorbital nerve in TN and TN+ Cur groups.Starting from 15 days after establishment of the model, 1.5 ml peanut oil was injected through a gastric tube into stomach twice a day (in the morning and at night) for 28 consecutive days in TN group, and curcumin 45 mg/kg (dissolved in 1.5 ml peanut oil) was injected through a gastric tube into stomach twice a day (in the morning and at night) for 28 consecutive days in TN+ Cur group.The cognitive function was assessed using Morris water maze test after the end of treatment.The escape latency, swimming speed, ratio of time of staying at the target quadrant, and the number of times the animals crossing the platform were recorded.The pathological changes in the hippocampal CA1 region were observed with an electron microscope.The ultrastructure of neurons, organelles and synapses in the hippocampal CA1 region was examined with a transmission electron microscope. Results There was no significant difference in the swimming speed between the three groups (P>0.05). Compared with Sham group, the escape latency on 1st-4th days was significantly prolonged, the ratio of time of staying at the target quadrant was decreased, and the number of times the animals crossing the platform was decreased in TN group, and the escape latency was significantly prolonged on 3rd and 4th days in TN+ Cur group (P<0.01). Compared with TN group, the escape latency on 2nd-4th days was significantly shortened, the ratio of time of staying at the target quadrant was increased, the number of times the animals crossing the platform was increased (P<0.01), and the pathological changes of hippocampal tissues were attenuated in TN+ Cur group. Conclusion Curcumin can improve cognitive dysfunction in a rat model of trigeminal neuralgia. Key words: CURCUMIN; Trigeminal neuralgia; Cognition disorders

Projeto MagLev Cobra - Levitação Supercondutora para Transporte Urbano

Neste trabalho apresentam-se as principais características do Projeto MagLev Cobra desenvolvido atualmente na Universidade Federal do Rio de Janeiro. Faz-se uma retrospectiva das técnicas de levitação magnética, destacando-se as vantagens da proposta aqui apresentada, comparativamente ao sistema roda-trilho e os VLT's (Veículos Leves sobre Trilhos). Apresenta-se a história do desenvolvimento do projeto. Detalhes técnicos, como a linha de imãs permanentes, os criostatos com supercondutores, o motor linear de tração, são discutidos. Finaliza-se destacando a sustentabilidade do projeto, tendo em vista seu impacto econômico, social e ambiental.

Hierarchical structure and mechanical properties of snake (Naja atra) and turtle (Ocadia sinensis) eggshells

After hundreds of million years of evolution, natural armors have evolved in various organisms, and has manifested in diverse forms such as eggshells, abalone shells, alligator osteoderms, turtle shells, and fish scales. Eggshells serve as multifunctional shields for successful embryogenesis, such as protection, moisture control and thermal regulation. Unlike calcareous avian eggshells which are brittle and hard, reptilians have leathery eggshells that are tough and flexible. Reptilian eggshells can withstand collision damages when laid in holes and dropped onto each other, and reduce abrasion caused by buried sand. In this study, we investigate structure and mechanical properties of eggshells of Taiwan cobra snake (Naja atra) and Chinese striped-neck turtle (Ocadia sinensis). From Acid Fuchsin Orange G (AFOG) staining and ATR-FTIR examination, we found that both eggshells are mainly composed of keratin. The mechanical properties of demineralized snake and turtle eggshells were evaluated by tensile and fracture tests and show distinctly difference. Turtle eggshells are relatively stiff and rigid, while snake eggshells behave as elastomers, which are highly extensible and reversible. The exceptional deformability (110–230% tensile strain) and toughness of snake eggshells are contributed by the wavy and random arrangement of keratin fibers as well as collagen layers. Multi-scale toughening mechanisms of snake eggshells were observed and elucidated, including crack deflection and twisting, fibers reorientation, sliding and bridging, inter-laminar shear effect, as well as the α–β phase transition of keratin. Inspirations from the structural and mechanical designs of reptilian eggshells may lead to the synthesis of tough, extensible, lightweight composites which could be further applied in the flexible devices, packaging and bio-medical fields. Statement of significanceAmniotic eggshells serve as multifunctional shields for successful embryogenesis. The avian eggshells have been extensively studied while there are very few studies on reptilian eggshells and most of them focused on mineralization and embryotic development. For the first time, the hierarchical structure and mechanical properties of snake and turtle eggshells are comprehensively and comparatively studied. Both snake and turtle eggshells are multilayer, hierarchically-structured composites consisting mainly of keratin yet their mechanical behaviors are distinctly different. Turtle eggshells are stiff and rigid, while snake eggshells are highly extensible (>200%) and reversible due to multiple deformation stages, phase transition of keratin and various toughening mechanisms. We believe that this study will make positive scientific impact and interest the broad and multidisciplinary readership.

Quantitation of Cytokine mRNA Expression in Cobra Snake Venom Stimulated PBMC of Horses Using Real-Time RT-PCR

Quantitation of Cytokine mRNA Expression in Cobra Snake Venom Stimulated PBMC of Horses Using Real-Time RT-PCR

Cobra venom proteome and glycome determined from individual snakes of Naja atra reveal medically important dynamic range and systematic geographic variation

Recent progress in snake venomics has shed much light on the intra-species variation among the toxins from different geographical regions and has provided important information for better snakebite management. Most previous reports on snake venomics were based on venoms pooled from different snakes. In this study, we present the proteomic and glycomic profiles of venoms from individual Naja atra snakes. The results reveal wide dynamic range of three-finger toxins. Systematic classification based on cardiotoxin (CTX-) profiles of A2/A4 and A6, respectively, allowed the identification of two putative subspecies of Taiwan cobra from the eastern and western regions. We also identified four major N-glycan moieties on cobra snake venom metalloproteinase on the bi-antennary glycan core. ELISA showed that these glycoproteins (<3%) could elicit much higher antibody response in antiserum when compared to other high-abundance cobra venom toxins such as small molecular weight CTXs (~60%). By removing these high-molecular weight glycoproteins from the immunogen, we demonstrated better protection than that achieved with conventional crude venom immunization in mice challenged by crude venom. We conclude that both intra-species and inter-individual variations of proteomic and glycomic profiles of snake venomics should be considered to provide better antivenomic approach for snakebite management. Biological significanceBased on the proteomic and glycomic profiles of venoms obtained from individual snakes, we demonstrated a surprisingly wide dynamic range and geographical variation of three-finger toxins in cobra venomics. This provides a reasonable explanation for the variable neutralization effects of antivenom treatment on victims suffering from cobra snakebite and suggests a simple and economic method to produce potent antivenom with better efficacy. Since two major venomic profiles with distinct dynamic ranges were observed for Taiwan cobra venoms isolated from the eastern and western regions, the current venomic profile should be used as a quality control for future production of antivenom in clinical applications.

Absence of inflammatory lung injury in rabbits challenged intravascularly with complement-derived chemotactic factors.

Systemic activation of the complement system results in the generation of chemotactic factors that have been suggested to play a role in the pathogenesis of inflammatory pulmonary diseases such as the adult respiratory distress syndrome. This led us to ask whether systemic complement activation by cobra venom factor (CVF) or intravascularly administered zymosan-activated rabbit plasma (ZAP) or rabbit C5a would result in lung injury. As had been described previously for CVF and ZAP, intravenously administered rabbit C5a also caused an acute neutropenia along with sequestration of neutrophils within the pulmonary vasculature. However, no significant lung inflammation as measured by neutrophil emigration or increased vascular permeability occurred with any of the three stimuli. Only when these agents were combined with anesthesia, surgical manipulation, and intubation did significant neutrophil emigration into alveoli occur, but again without any change in vascular permeability. After administration of ZAP, a decrease in dynamic compliance and an increase in pulmonary resistance as well as a transient period of hypoxemia occurred that was not observed after CVF or rabbit C5a treatment. Thus, our studies suggest that changes in lung function after ZAP instillation may not represent changes from complement activation alone in that they are not reproduced with CVF or rabbit C5a. We conclude that complement activation, as an isolated event, may be an insufficient insult in the lung to produce significant lung injury.

Substrate zymography of snake venom components depicting the activity of matrix metalloprotease

Introduction: Matrix metalloproteases (MMPs) are a group of endopeptidases belonging to a superfamily of zinc protease enzymes that have the potential to degrade the extracellular matrix (ECM). The loss of its activity could lead to very serious alterations in the human body, causing several conditions such as arthritis and aneurisms. Previous studies indicate that snake venom contains snake venom metalloprotease (SVMP), which might possess similar activity to MMPs. In these experiments, it was shown that Malaysian cobra SVMP is capable of gelatinolytic activity, which was demonstrated using gelatine as substrate in sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) (zymography). Objectives: To investigate the activity of Malaysian cobra SVMP using zymography. Materials and methods: Protein was extracted from cobra snake venom using fast protein liquid chromatography then, after purification, the fractions were collected and run on SDS-PAGE to visualize the protein of interest, which was then desalted and concentrated using dialysis and ready to be used for zymography to demonstrate the activity of SVMP. Results: In the detection of the SVMP on SDS-PAGE, the molecular weight was observed around 55kDa. A possible dimer was also seen at 110kDa. Gelatinase activity of SVMP was clearly revealed on the gelatine gel that was used in zymography. Conclusions: Having found the gelatinolytic activity of SVMP the future direction of this protein can be tested against the tumour environment to see the potential use of SVMP’ s. Acknowledgements: Al-Imam Muhammad ibn Saud Islamic University for their support, International Medical University for using their laboratories, and Dr Fabian Davamani for supervising this project.