Abu Reza

Unusual accelerated rate of deletions and insertions in toxin genes in the venom glands of the pygmy copperhead (Austrelaps labialis) from kangaroo island

BackgroundToxin profiling helps in cataloguing the toxin present in the venom as well as in searching for novel toxins. The former helps in understanding potential pharmacological profile of the venom and evolution of toxins, while the latter contributes to understanding of novel mechanisms of toxicity and provide new research tools or prototypes of therapeutic agents.ResultsThe pygmy copperhead (Austrelaps labialis) is one of the less studied species. In this present study, an attempt has been made to describe the toxin profile of A. labialis from Kangaroo Island using the cDNA library of its venom glands. We sequenced 658 clones which represent the common families of toxin genes present in snake venom. They include (a) putative long-chain and short-chain neurotoxins, (b) phospholipase A2, (c) Kunitz-type protease inhibitor, (d) CRISPs, (e) C-type lectins and (f) Metalloproteases. In addition, we have also identified a novel protein with two Kunitz-type domains in tandem similar to bikunin.ConclusionInterestingly, the cDNA library reveals that most of the toxin families (17 out of 43 toxin genes; ~40%) have truncated transcripts due to insertion or deletion of nucleotides. These truncated products might not be functionally active proteins. However, cellular trancripts from the same venom glands are not affected. This unusual higher rate of deletion and insertion of nucleotide in toxin genes may be responsible for the lower toxicity of A. labialis venom of Kangroo Island and have significant effect on evolution of toxin genes.

Gene duplication of coagulation factor V and origin of venom prothrombin activator in Pseudonaja textilis snake

The origin and evolution of venom toxins is a mystery that has evoked much interest. We have recently shown that pseutarin C, a prothrombin activator from Pseudonaja textilis venom, is structurally and functionally similar to mammalian coagulation factor Xa-factor Va complex. Its catalytic subunit is homologous to factor Xa while the nonenzymatic subunit is homologous to factor Va. P. textilis therefore has two parallel prothrombin activator systems: one expressed in its venom gland as a toxin and the other expressed in its liver and released into its plasma as a haemostatic factor. Here we report the complete amino acid sequence of factor V (FV) from its liver determined by cDNA cloning and sequencing. The liver FV shows 96% identity to pseutarin C nonenzymatic subunit. Most of the functional sites involved in its interaction with factor Xa and prothrombin are conserved. However, many potential sites of post-translational modifications and one critical cleavage site for activated protein C are different. The absence of the latter cleavage site makes pseutarin C nonenzymatic subunit resistant to inactivation and enhances its potential as an excellent toxin. By PCR and real-time quantitative analysis, we show that pseutarin C nonenzymatic subunit gene is expressed specifically in the venom gland at approximately 280 fold higher than that of FV gene in liver. These two are thus encoded by two separate genes that express in a highly tissue-specific manner. Our results imply that the gene encoding pseutarin C nonenzymatic subunit was derived by the duplication of plasma FV gene and they have evolved to perform distinct functions.

Interrogating the Venom of the Viperid Snake Sistrurus catenatus edwardsii by a Combined Approach of Electrospray and MALDI Mass Spectrometry.

The complete sequence characterization of snake venom proteins by mass spectrometry is rather challenging due to the presence of multiple isoforms from different protein families. In the present study, we investigated the tryptic digest of the venom of the viperid snake Sistrurus catenatus edwardsii by a combined approach of liquid chromatography coupled to either electrospray (online) or MALDI (offline) mass spectrometry. These different ionization techniques proved to be complementary allowing the identification a great variety of isoforms of diverse snake venom protein families, as evidenced by the detection of the corresponding unique peptides. For example, ten out of eleven predicted isoforms of serine proteinases of the venom of S. c. edwardsii were distinguished using this approach. Moreover, snake venom protein families not encountered in a previous transcriptome study of the venom gland of this snake were identified. In essence, our results support the notion that complementary ionization techniques of mass spectrometry allow for the detection of even subtle sequence differences of snake venom proteins, which is fundamental for future structure-function relationship and possible drug design studies.

The Antioxidative Fraction of White Mulberry Induces Apoptosis through Regulation of p53 and NFκB in EAC Cells.

In this study, the antioxidative fraction of white mulberry (Morus alba) was found to have an apotogenic effect on Ehrlich’s ascites carcinoma cell-induced mice (EAC mice) that correlate with upregulated p53 and downregulated NFκB signaling. The antioxidant activities and polyphenolic contents of various mulberry fractions were evaluated by spectrophotometry and the ethyl acetate fraction (EAF) was selected for further analysis. Strikingly, the EAF caused 70.20% tumor growth inhibition with S-phase cell cycle arrest, normalized blood parameters including red/white blood cell counts and suppressed the tumor weight of EAC mice compared with untreated controls. Fluorescence microscopy analysis of EAF-treated EAC cells revealed DNA fragmentation, cell shrinkage, and plasma membrane blebbing. These characteristic morphological features of apoptosis influenced us to further investigate pro- and anti-apoptotic signals in EAF-treated EAC mice. Interestingly, apoptosis correlated with the upregulation of p53 and its target genes PARP-1 and Bax, and also with the down-regulation of NFκB and its target genes Bcl-2 and Bcl-xL. Our results suggest that the tumor- suppressive effect of the antioxidative fraction of white mulberry is likely due to apoptosis mediated by p53 and NFκB signaling.

ISOB: A Database of Indigenous Snake Species of Bangladesh with respective known venom composition

At present there is no well structured database available for the venomous snakes and venom composition of snakes in the world although venom has immense importance in biomedical research. Searching for a specific venom component from NCBI, PDB or public databases is troublesome, because they contain huge amount of data entries. Therefore, we created a database named “ISOB” which is a web accessible unique secondary database that represents the first online available bioinformatics resource showing venom composition of snakes. This database provides a comprehensive overview of seventy-eight indigenous snake species covering description of snakes supplemented with structural information of the relevant individual available venom proteins. We strongly believe that this database will contribute significantly in the field of bioinformatics, environmental research, proteomics, drug development and rationale drug designing. Availability The database is freely available at http://www.snakebd.com/

Extraction and optimisation of red pigment production as secondary metabolites from Talaromyces verruculosus and its potential use in textile industries

ABSTRACT Textile dyes and effluents are considered as one of the worst polluters of our priceless water sources and soils. New sources of natural pigments are getting particular research interests due to the toxicity produced by synthetic colouring agents. Plant sources are being explored extensively for natural pigments but inadequate yield of those sources hampered the progression. Apart from the enormous antibacterial applications, fungi may provide a readily available alternative source of natural pigments. Here, we isolated a fungal strain from spoiled mango which is capable of producing pigments suitable for textile dyeing. The spoiled mangoes were selected as a source of different fungi. Among them one particular fungal isolate was selected for its visible production of secondary metabolites. Molecular identification using internal transcribed spacer sequencing revealed the fungi as Talaromyces verruculosus strain. The growth and pigment production of the fungi was optimised to obtain highest yield. Extracted pigment was applied to cotton fabric following a standard dyeing procedure for natural pigment. Adequate colour yield and negative cytotoxicity result suggested that the fungi source of pigment could be a potential replacement for hazardous synthetic dyes.

Remediation of Chromium Toxicity Through Exogenous Salicylic Acid in Rice ( Oryza sativa L.)

This work investigates whether and how salicylic acid (SA) alleviates chromium (Cr) toxicity in rice. Addition of SA under Cr stress markedly increased growth parameters, total protein content, and membrane stability but reduced the concentration and translocation of Cr in shoots but not in roots, suggesting that SA does have critical roles in Cr detoxification associated with Cr sequestration in roots. Further, Fe along with the expression of two Fe transporters (OsIRT1, OsNRAMP1) showed no significant changes in roots due to SA supplementation under Cr stress, indicating that regulation of Fe uptake is not involved in Cr reduction in rice plants through SA. At molecular level, OsPCS1 (phytochelatin synthase) and OsMT1 (metallothionein) and OsHMA3 (P-type ATPase 3) transcripts significantly upregulated following SA supplementation under Cr stress, suggesting that these chelating agents may bind to Cr leading to elevated Cr retention in roots. Furthermore, increased CAT, POD, SOD, and GR leading to decreased H2O2 along with elevated metabolites (cysteine, methionine, glutathione, proline, ascorbic acid) in roots implies active involvement of ROS scavenging and plays partial role in SA-mediated alleviation of Cr toxicity in rice plants. These findings will be useful for bioremediation of Cr toxicity in rice and other crops.

PROTHROMBIN ACTIVATORS FROM AUSTRALIAN SNAKES

Blood coagulation is a highly synchronized event of sequential activation reactions of several coagulation factors, and prothrombin activation is in the center of this cascade. In the physiological system, activated coagulation factor X (FXa) forms a complex (prothrombinase complex) with factor Va, Ca2+, and phospholipids and converts prothrombin to thrombin. Several exogenous prothrombin activators, particularly groups C and D from the Australian snake venoms, also activate prothrombin in a similar fashion. Recent studies have shown that they are structurally and functionally similar to the blood coagulation factors. Group C prothrombin activators resemble FVa-FXa complex, whereas group D prothrombin activators are similar to FXa. Thus, these snakes possess two parallel prothrombin activating systems, one in their venom and the other in their blood. These closely related proteins have distinctly different physiological roles. In this review, we will discuss the structural characteristics and evolution of the venom prothrombin activators and blood coagulation factors from Australian elapids.