Characterization of Major Zinc Containing Myonecrotic and Procoagulant Metalloprotease `Malabarin' from Non Lethal Trimeresurus malabaricus Snake Venom with Thrombin Like Activity: Its Neutralization by Chelating Agents

A major myonecrotic zinc containing metalloprotease `malabarin' with thrombin like activity was purified by the combination of gel permeation and anion exchange chromatography from T. malabaricus snake venom. MALDI-TOF analysis of malabarin indicated a molecular mass of 45.76 kDa and its N-terminal sequence was found to be Ile-Ile-Leu-Pro(Leu)-Ile-Gly-Val-Ile-Leu(Glu)-Thr-Thr. Atomic absorption spectral analysis of malabarin raveled the association of zinc metal ion. Malabarin is not lethal when injected i.p. or i.m. but causes extensive hemorrhage and degradation of muscle tissue within 24 hours. Sections of muscle tissue under light microscope revealed hemorrhage and congestion of blood vessel during initial stage followed by extensive muscle fiber necrosis with elevated levels of serum creatine kinase and lactate dehydrogenase activity. Malabarin also exhibited strong procoagulant action and its procoagulant action is due to thrombin like activity; it hydrolyzes fibrinogen to form fibrin clot. The enzyme preferentially hydrolyzes A alpha followed by B beta subunits of fibrinogen from the N-terminal region and the released products were identified as fibrinopeptide A and fibrinopeptide B by MALDI. The myonecrotic, fibrinogenolytic and subsequent procoagulant activities of malabarin was neutralized by specific metalloprotease inhibitors such as EDTA, EGTA and 1, 10-phenanthroline but not by PMSF a specific serine protease inhibitor. Since there is no antivenom available to neutralize local toxicity caused by T. malabaricus snakebite, EDTA chelation therapy may have more clinical relevance over conventional treatment.

The sequence and structure of snake gourd (Trichosanthes anguina) seed lectin, a three-chain nontoxic homologue of type II RIPs

The sequence and structure of snake gourd seed lectin (SGSL), a nontoxic homologue of type II ribosome-inactivating proteins (RIPs), have been determined by mass spectrometry and X-ray crystallography, respectively. As in type II RIPs, the molecule consists of a lectin chain made up of two beta-trefoil domains. The catalytic chain, which is connected through a disulfide bridge to the lectin chain in type II RIPs, is cleaved into two in SGSL. However, the integrity of the three-dimensional structure of the catalytic component of the molecule is preserved. This is the first time that a three-chain RIP or RIP homologue has been observed. A thorough examination of the sequence and structure of the protein and of its interactions with the bound methyl-alpha-galactose indicate that the nontoxicity of SGSL results from a combination of changes in the catalytic and the carbohydrate-binding sites. Detailed analyses of the sequences of type II RIPs of known structure and their homologues with unknown structure provide valuable insights into the evolution of this class of proteins. They also indicate some variability in carbohydrate-binding sites, which appears to contribute to the different levels of toxicity exhibited by lectins from various sources.

FURTHER RECORDS OF THE SINDH AWL-HEADED SNAKE, Lytorhynchus paradoxus (GUNTHER, 1875), FROM INDIA WITH NOTES ON ITS HABITAT AND NATURAL HISTORY

This paper presents additional distributional records of the Sindh awl-headed snake Lytorhynchus paradoxus from India, along with scale counts, measurements and natural history observations of this poorly known species.

ELISA for the detection of venoms from four medically important snakes of India

A double antibody sandwich enzyme linked immunosorbent assay (ELISA) was developed to detect Echis carinatus venom in various organs (brain, heart, lungs, liver, spleen and kidneys) as well as tissue at the site of injection of mice, at various time intervals (1, 6, 12, 18, 24 h and 12 h intervals up to 72 h) after death. The assay could detect E. carinatus venom levels up to 2.5 ng/ml of tissue homogenate and the venom was detected up to 72 h after death. A highly sensitive and species-specific avidin-biotin microtitre ELISA was also developed to detect venoms of four medically important Indian snakes (Bungarus caeruleus, Naja naja, E. carinatus and Daboia russelli russelli) in autopsy specimens of human victims of snake bite. The assay could detect venom levels as low as 100 pg/ml of tissue homogenate. Venoms were detected in brain, heart, lungs, liver, spleen, kidneys, tissue at the bite area and postmortem blood. In all 12 human victim cadavers tested the culprit species were identified. As observed in mice, tissue at the site of bite area showed the highest concentration of venom and the brain showed the least. Moderate amounts of venoms were found in liver, spleen, kidneys, heart and lungs. Development of a simple, rapid and species-specific diagnostic kit based on this ELISA technique useful to clinicians is discussed.

A real-time algorithm for simulation of flexible objects and hyper-redundant manipulators

Flexible objects such as a rope or snake move in a way such that their axial length remains almost constant. To simulate the motion of such an object, one strategy is to discretize the object into large number of small rigid links connected by joints. However, the resulting discretised system is highly redundant and the joint rotations for a desired Cartesian motion of any point on the object cannot be solved uniquely. In this paper, we revisit an algorithm, based on the classical tractrix curve, to resolve the redundancy in such hyper-redundant systems. For a desired motion of the `head' of a link, the `tail' is moved along a tractrix, and recursively all links of the discretised objects are moved along different tractrix curves. The algorithm is illustrated by simulations of a moving snake, tying of knots with a rope and a solution of the inverse kinematics of a planar hyper-redundant manipulator. The simulations show that the tractrix based algorithm leads to a more `natural' motion since the motion is distributed uniformly along the entire object with the displacements diminishing from the `head' to the `tail'.

Lipid composition of the silkworm Bombyx mori L

The constituents of silkworm fat were studied in detail. An unsaturated fat with a high concentration of phospholipid was generally observed. Its iodine value increased during metamorphosis. The free fatty acid concentration likewise increased from the spinning larvae to the moth stage. Analyses of silkworm organs revealed that the fat body had the most fat and the least free fatty acids, whereas haemolymph contained the least fat. Silk glands contained the maximum phospholipid percentage. Stearic acid predominated in those tissues that had a high percentage of phospholipid. Stearic acid was the predominant saturated fatty acid in both the phospholipids and lecithin, and it accounted for 35–50 per cent of the free fatty acids of all the tissues. Q10 was the ubiquinone present; also found were ubichromenol and tocopherol. Results show that silkworm sterol may be cholesterol. Intestines contained the maximum quantities of sterol, ubiquinone, ubichromenol, and tocopherol. The composition of silkworm phospholipids varies considerably from those of other insects, but lecithin is comparable in its composition with lecithins of other animals. The phospholipids had with them a highly complexed protein along with a polysaccharide. In experiments with snake venoms unsaturated fatty acids were found to be predominantly released from silkworm lecithin.

Multiple contour extraction from graylevel images using an artificial neural network

For active contour modeling (ACM), we propose a novel self-organizing map (SOM)-based approach, called the batch-SOM (BSOM), that attempts to integrate the advantages of SOM- and snake-based ACMs in order to extract the desired contours from images. We employ feature points, in the form of ail edge-map (as obtained from a standard edge-detection operation), to guide the contour (as in the case of SOM-based ACMs) along with the gradient and intensity variations in a local region to ensure that the contour does not "leak" into the object boundary in case of faulty feature points (weak or broken edges). In contrast with the snake-based ACMs, however, we do not use an explicit energy functional (based on gradient or intensity) for controlling the contour movement. We extend the BSOM to handle extraction of contours of multiple objects, by splitting a single contour into as many subcontours as the objects in the image. The BSOM and its extended version are tested on synthetic binary and gray-level images with both single and multiple objects. We also demonstrate the efficacy of the BSOM on images of objects having both convex and nonconvex boundaries. The results demonstrate the superiority of the BSOM over others. Finally, we analyze the limitations of the BSOM.

Redundancy resolution using a tractrix and its application to real-time simulations of hyper-redundant manipulators, snakes and tying of knots

To realistically simulate the motion of flexible objects such as ropes, strings, snakes, or human hair,one strategy is to discretise the object into a large number of small rigid links connected by rotary or spherical joints. The discretised system is highly redundant and the rotations at the joints (or the motion of the other links) for a desired Cartesian motion of the end of a link cannot be solved uniquely. In this paper, we propose a novel strategy to resolve the redundancy in such hyper-redundant systems.We make use of the classical tractrix curve and its attractive features. For a desired Cartesian motion of the `head'of a link, the `tail' of the link is moved according to a tractrix,and recursively all links of the discretised objects are moved along different tractrix curves. We show that the use of a tractrix curve leads to a more `natural' motion of the entire object since the motion is distributed uniformly along the entire object with the displacements tending to diminish from the `head' to the `tail'. We also show that the computation of the motion of the links can be done in real time since it involves evaluation of simple algebraic, trigonometric and hyperbolic functions. The strategy is illustrated by simulations of a snake, tying of knots with a rope and a solution of the inverse kinematics of a planar hyper-redundant manipulator.

Snakes with Ellipse-Reproducing Property

We present a new class of continuously defined parametric snakes using a special kind of exponential splines as basis functions. We have enforced our bases to have the shortestpossible support subject to some design constraints to maximize efficiency. While the resulting snakes are versatile enough to provide a good approximation of any closed curve in the plane, their most important feature is the fact that they admit ellipses within their span. Thus, they can perfectly generate circular and elliptical shapes. These features are appropriate to delineate cross sections of cylindrical-like conduits and to outline blob-like objects. We address the implementation details and illustrate the capabilities of our snake with synthetic and real data.

Snakes With an Ellipse-Reproducing Property

We present a new class of continuously defined parametric snakes using a special kind of exponential splines as basis functions. We have enforced our bases to have the shortest possible support subject to some design constraints to maximize efficiency. While the resulting snakes are versatile enough to provide a good approximation of any closed curve in the plane, their most important feature is the fact that they admit ellipses within their span. Thus, they can perfectly generate circular and elliptical shapes. These features are appropriate to delineate cross sections of cylindrical-like conduits and to outline bloblike objects. We address the implementation details and illustrate the capabilities of our snake with synthetic and real data.

Active-contour-based automated image quantitation techniques for Western Blot analysis

Western Blot analysis is an analytical technique used in Molecular Biology, Biochemistry, Immunogenetics and other Molecular Biology studies to separate proteins by electrophoresis. The procedure results in images containing nearly rectangular-shaped blots. In this paper, we address the problem of quantitation of the blots using automated image processing techniques. We formulate a special active contour (or snake) called Oblong, which locks on to rectangular shaped objects. Oblongs depend on five free parameters, which is also the minimum number of parameters required for a unique characterization. Unlike many snake formulations, Oblongs do not require explicit gradient computations and therefore the optimization is carried out fast. The performance of Oblongs is assessed on synthesized data and Western Blot Analysis images.

Forgotten records of Chrysopelea taprobanica Smith, 1943 (Squamata: Colubridae) from India

The colubrid snake Chrysopelea taprobanica Smith, 1943 was described from a holotype from Kanthali (= Kantalai) and paratypes from Kurunegala, both localities in Sri Lanka (formerly Ceylon) (Smith 1943). Since its description, literature pertaining to Sri Lankan snake fauna considered this taxon to be endemic to the island (Taylor 1950, Deraniyagala 1955, de Silva 1980, de Silva 1990, Somaweera 2004, Somaweera 2006, de Silva 2009, Pyron et al. 2013). In addition, earlier efforts on the Indian peninsula (e.g. Das 1994, 1997, Das 2003, Whitaker & Captain 2004, Aengals et al. 2012) and global data compilations (e.g. Wallach et al. 2014, Uetz & Hošek 2015) did not identify any record from mainland India until Guptha et al. (2015) recorded a specimen (voucher BLT 076 housed at Bio-Lab of Seshachalam Hills, Tirupathi, India) in the dry deciduous forest of Chamala, Seshachalam Biosphere Reserve in Andhra Pradesh, India in November 2013. Guptha et al. (2015) further mentioned an individual previously photographed in 2000 at Rishi Valley, Andhra Pradesh, but with no voucher specimen collected. Guptha’s record, assumed to be the first confirmed record of C. taprobanica in India, is noteworthy as it results in a large range extension, from northern Sri Lanka to eastern India with an Euclidean distance of over 400 km, as well as a change of status, i.e., species not endemic to Sri Lanka. However, at least three little-known previous records of this species from India evaded most literature and were overlooked by the researchers including ourselves.