Neurotoxic activity and ultrastructural changes in muscles caused by the brown widow spider Latrodectus geometricus venom

Brown widow spider (Latrodectus geometricus) venom (BrWSV) produces few local lesions and intense systemic reactions such as cramps, harsh muscle pains, nausea, vomiting and hypertension. Approximately 16 protein bands under reducing conditions and ~ 14 bands under non-reducing conditions on a 12.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis were observed. Neurotoxic clinical manifestations were confirmed in vivo, while proteolytic activity was demonstrated on gelatine film. Severe ultrastructural damages in mice skeletal muscles were observed at 3, 6, 12 and 24 h postinjection with at total of 45 µg of venom protein. Infiltration of eosinophils and ruptures of the cellular membranes were observed in the muscles along with swelling of the nuclear cover and interruption of the collagen periodicity. Altered mitochondrias and autophage vacuoles, nuclear indentation and mitochondria without cristae, slight increment of intermyofibrillar and subsarcolemic spaces and myelinic figures formation were also observed. In the capillary, endothelial membrane unfolding into the lumen was noticed; along with myelinic figures compatible with a toxic myopathy. Swollen sarcotubular systems with lysis of membrane, intense mitochondria autophagia and areas without pinocytic vesicles were observed. Swollen mitochondria surrounded by necrotic areas, myofibrillar disorganization and big vacuolas of the sarcotubular system, degenerated mitochondrium with formation of myelinic figure was seen. Glycogenosomes with small particulate, muscle type glycogen was noticed. Autophagic vacuole (autophagolysosomes) and necrotic areas were also noticed. These damages may be due to interactive effects of the multifactorial action of venom components. However, Latrodectus geometricus venom molecules may also be utilized as neuro therapeutic tools, as they affect neuronal activities with high affinity and selectivity. To our knowledge, the present study is the first ultrastructural report in the literature of muscle injuries and neurological and proteolytic activities caused by BrWSV.

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Activities against hemostatic proteins and adrenal gland ultrastructural changes caused by the brown widow spider Latrodectus geometricus (Araneae: Theridiidae) venom

Comparative Biochemistry and Physiology Part C Toxicology & Pharmacology ◽

10.1016/j.cbpc.2009.09.005 ◽

2010 ◽

Vol 151 (1) ◽

pp. 113-121 ◽

Cited By ~ 5

Author(s):

Belsy Guerrero ◽

Hector J. Finol ◽

Matias Reyes-Lugo ◽

Ana M. Salazar ◽

Elda E. Sánchez ◽

...

Keyword(s):

Adrenal Gland ◽

Ultrastructural Changes ◽

Latrodectus Geometricus ◽

Brown Widow Spider ◽

Widow Spider

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Partial proteomic analysis of brown widow spider (Latrodectus geometricus) venom to determine the biological activities

Toxicon X ◽

10.1016/j.toxcx.2020.100062 ◽

2020 ◽

Vol 8 ◽

pp. 100062

Author(s):

Pornsawan Khamtorn ◽

Prapenpuksiri Rungsa ◽

Nisachon Jangpromma ◽

Sompong Klaynongsruang ◽

Jureerut Daduang ◽

...

Keyword(s):

Proteomic Analysis ◽

Biological Activities ◽

Latrodectus Geometricus ◽

Brown Widow Spider ◽

Widow Spider

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Atrial natriuretic peptide receptors in renal papilla of DOCA-salt hypertensive rats

AJP Renal Physiology ◽

10.1152/ajprenal.1990.259.1.f130 ◽

1990 ◽

Vol 259 (1) ◽

pp. F130-F137

Author(s):

E. Nuglozeh ◽

G. Gauquelin ◽

R. Garcia ◽

J. Tremblay ◽

E. L. Schiffrin

Keyword(s):

Atrial Natriuretic Peptide ◽

Natriuretic Peptide ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Plasma Renin ◽

Renal Papilla ◽

Hypertensive Rats ◽

Reducing Conditions ◽

Anp Receptors ◽

Atrial Natriuretic

The receptor for atrial natriuretic peptide (ANP) in the rat renal papilla was characterized pharmacologically. After solubilization and irreversible binding with disuccinimidylsuberate, it was shown on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) to be made of a single peptide of 125 kDa. The regulation of the renal papillary ANP receptor was studied in deoxycorticosterone acetate (DOCA)-salt hypertensive rats. DOCA-salt rats had suppressed plasma renin activity and increased plasma ANP concentrations (408 +/- 35 vs. 133 +/- 12 pg/ml in uninephrectomized controls, P less than 0.01). The renal papilla was hypertrophied in DOCA-salt hypertensive rats (93 +/- 1 vs. 52 +/- 1 mg, P less than 0.01). The density of ANP sites in the papilla was significantly higher in DOCA-salt rats (141 +/- 31 fmol/papilla) than in controls (34 +/- 8 fmol/papilla, P less than 0.01). Affinity of sites in DOCA-salt rats and controls was similar. The production of guanosine 3',5'-cyclic monophosphate (cGMP) in renal papilla in response to ANP was significantly higher in DOCA-salt rats. In contrast to the renal papillary ANP receptor, acid-washed vascular and glomerular ANP sites were significantly decreased in density in DOCA-salt hypertensive rats. In blood vessels and glomeruli, both the high- and low-molecular mass receptor (as detected on SDS-PAGE under reducing conditions) was proportionately decreased in density in DOCA-salt hypertensive rats. The present results suggest that an increased number of ANP receptors and exaggerated cGMP response to ANP in the renal papilla may underlie the increased natriuretic responsiveness of the kidney to ANP in DOCA-salt hypertensive rats.

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Glutathione-s-transferase is an important antigen in the eel nematode Anguillicola crassus

Journal of Helminthology ◽

10.1017/s0022149x00016138 ◽

1997 ◽

Vol 71 (4) ◽

pp. 319-324 ◽

Cited By ~ 10

Author(s):

M.E. Nielsen ◽

K. Buchmann

Keyword(s):

Specific Antibody ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Anguilla Anguilla ◽

European Eel ◽

Glutathione S Transferase ◽

Elisa System ◽

Humoral Immune ◽

Anguillicola Crassus ◽

Reducing Conditions

AbstractDifferent organs and secretions/excretions of the swimbladder parasite, Anguillicola crassus (Nematoda), were tested for the presence of antigens to the humoral immune response previously detected in the European eel, Anguilla anguilla. Proteins from different fractions of Anguillicola crassus were separated using SDS–PAGE (sodium-dodecyl-sulphate polyacrylamide-gel electrophoresis) under reducing conditions and electroblotted onto nitrocellulose membranes. Infected eels showed a specific antibody response to a 43 kDa antigen in the cuticle and towards two gonad antigens around 34 and 43 kDa. In protein released from the worms, two secretory/excretory antigens of approximately 28 kDa were found. The secretion/excretion rate of protein from the parasite to the surroundings was determined. Subsequently, an ELISA system was established applying these antigens as the first layer of coating. Furthermore, antigens from Anguillicola crassus were examined for the presence of glutathione-s-transferase (GST) using a specific antibody against GST. The antigens were found to be subunits of GST.

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Immunoprecipitation of the lutropin receptor. Loss of receptor molecules during down-regulation

Biochemical Journal ◽

10.1042/bj2240467 ◽

1984 ◽

Vol 224 (2) ◽

pp. 467-471 ◽

Cited By ~ 4

Author(s):

M K Metsikkö ◽

H J Rajaniemi

Keyword(s):

Gamma Globulin ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Down Regulation ◽

Gonadotropin Receptor ◽

Reducing Conditions ◽

Lutropin Receptor ◽

Rat Ovary ◽

Triton X ◽

Receptor Antibodies

Specific anti-(lutropin receptor) antibodies were produced by immunizing rabbits with lutropin receptor purified from pseudopregnant rat ovary. The anti-receptor serum at 1:100 dilution together with anti-(rabbit gamma-globulin) serum immunoprecipitated 70% of 3H-labelled, purified lutropin receptor and 42% of 125I-chorio-gonadotropin-receptor complex. The antiserum inhibited hormone binding to rat ovarian particles. Pseudopregnant rat ovarian particles were labelled with periodate/NaB3H4 and solubilized with Triton X-100. The Triton X-100 extract was subjected to immunoprecipitation using the anti-receptor serum. When the immunoprecipitate was dissolved and analysed by polyacrylamide-gel electrophoresis in sodium dodecyl sulphate under reducing conditions followed by fluorography, a receptor polypeptide with an apparent Mr 95000 was detected. A receptor down-regulating dose of choriogonadotropin was injected into pseudopregnant rats and their ovaries were removed and homogenized 4 days later, and analysed for immunoprecipitable receptors as above. No receptor molecules were found. Accordingly, the lutropin receptor molecules actually disappear rather than merely become masked from hormone during homologous down-regulation.

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Albumin associated with purified pig lymphocyte plasma membrane

Biochemical Journal ◽

10.1042/bj1920049 ◽

1980 ◽

Vol 192 (1) ◽

pp. 49-57 ◽

Cited By ~ 23

Author(s):

M J Owen ◽

B H Barber ◽

R A Faulkes ◽

M J Crumpton

Keyword(s):

Plasma Membrane ◽

Serum Albumin ◽

Sodium Dodecyl Sulphate ◽

Gel Electrophoresis ◽

Membrane Fraction ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Reducing Conditions ◽

Isoelectric Points ◽

Pig Serum

Plasma-membrane preparations purified from pig lymphocytes contained a major polypeptide component of mol.wt. about 68 000. This component was identified as pig albumin by the following comparisons with authentic pig serum albumin: (a) co-migration when analysed by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis under reducing and non-reducing conditions; (b) identical isoelectric points; (c) similar “fingerprints” of arginine-containing tryptic peptides; (d) reactivity with anti-(pig albumin) serum. The albumin was bound tightly to the plasma membrane. Biosynthetic labelling of pig lymphocytes under a variety of conditions failed to provide evidence that albumin was synthesized by lymphocytes, suggesting that the plasma-membrane-associated albumin was of extraneous origin. Radiolabelled pig serum albumin, however, failed to bind to the plasma-membrane fraction when added before cell disruption. Although lymphocyte plasma membrane preparations from other species possessed a polypeptide of about 68 000 mol.wt., this was judged not to be albumin on the basis of electrophoretic mobility under non-reducing conditions; also, no polypeptide was precipitated by anti-albumin sera. It is concluded that pig lymphocyte plasma-membrane preparations possess albumin which, although firmly attached, was probably of extraneous origin. This association appeared not to be common to lymphocytes from other species.

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The brown widow spider Latrodectus geometricus C. L. Koch, 1841, in southern California

The Pan-Pacific Entomologist ◽

10.3956/2008-07.1 ◽

2008 ◽

Vol 84 (4) ◽

pp. 344-349 ◽

Cited By ~ 17

Author(s):

Leonard S. Vincent ◽

Richard S. Vetter ◽

William J. Wrenn ◽

Janet K. Kempf ◽

James E. Berrian

Keyword(s):

Southern California ◽

Latrodectus Geometricus ◽

Brown Widow Spider ◽

Widow Spider

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Isolation and Preliminary Characterisation of Active B-Chain of Recombinant Tissue-Type Plasminogen Activator

Thrombosis and Haemostasis ◽

10.1055/s-0038-1661455 ◽

1986 ◽

Vol 55 (01) ◽

pp. 094-097 ◽

Cited By ~ 27

Author(s):

I Dodd ◽

R Fears ◽

J H Robinson

Keyword(s):

Plasminogen Activator ◽

Specific Activity ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Tissue Type ◽

Tissue Type Plasminogen Activator ◽

Reducing Conditions ◽

Type Plasminogen Activator ◽

Urokinase Type Plasminogen Activator ◽

Single Polypeptide

SummaryPurified 2-chain recombinant tissue-type plasminogen activator (t-PA) was reduced under mild conditions - 10 mM dithiothreitol/ 5° C/1.5 h - and the two chains were separated by chromatography on lysine Sepharose. The t-PA B chain was fully active as determined by its activity towards the chromogenic substrate S-2288 (H-D-ile-pro-arg p-nitroanilide). Analysis by sodium dodecyl sulphate polyacrylamide gel electrophoresis under reducing or non-reducing conditions revealed a single polypeptide at Mr = 35,000 or 29,000 respectively. In addition, under non-reducing conditions a fibrinolytic band at apparent Mr = 29,000 was present after fibrin zymography. The N-terminal sequence was confirmed as ile-lys-gly. The t-PA B chain had a specific amidolytic activity, using S-2288, of 170,000 to 210,000 SU/mg protein. (This compares to a specific activity of the native 2-chain t-PA of 170,000 SU/mg). It resembles urokinase-type plasminogen activator in its inability to be stimulated by fibrin and its dose response on human fibrin plates. However, t-PA B-chain was stimulated to almost the same extent as t-PA by poly-D-lysine. The isoelectric points, at pH 5.6 and 5.7, fall outside the range generally quoted for t-PA preparations (pH 7.8-8.8).

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Persistent Bacteremia Due to Coagulase-Negative Staphylococci in Low Birth Weight Neonates

PEDIATRICS ◽

10.1542/peds.84.6.977 ◽

1989 ◽

Vol 84 (6) ◽

pp. 977-985

Author(s):

Christian C. Patrick ◽

Sheldon L. Kaplan ◽

Carol J. Baker ◽

Joseph T. Parisi ◽

Edward O. Mason

Keyword(s):

Birth Weight ◽

Low Birth Weight ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Clinical Manifestations ◽

Blood Cultures ◽

Coagulase Negative Staphylococci ◽

Low Birth Weight Infants ◽

Persistent Bacteremia ◽

Intravascular Catheters

During a 6-month period in 1987, 13 low birth weight neonates without indwelling central intravascular catheters had persistent (positive blood cultures for ≥6 days) coagulase-negative staphylococcal bacteremia despite adequate antibiotic therapy. Daily blood cultures remained persistently positive for a mean of 13 days (range 6 to 25 days). This group of infants was compared with other low birth weight infants with similar birth weights and nonpersistent coagulase-negative staphylococcal bacteremia, defined as two or more positive blood cultures accompanied by supporting clinical manifestations of sepsis. During this period, coagulase-negative staphylococcal represented 29% of all bacteremias, and 33% of coagulase-negative staphylococcal bacteremias were persistent. Other than soft tissue abscesses, none of the infants with persistent coagulase-negative staphylococcal bacteremia had a defined focus of infection. Abdominal distention (P= .001) and thrombocytopenia (P<.03) occurred significantly more frequently in the patients with persistent coagulase-negative staphylococcal bacteremia than in those with nonpersistent bacteremia. Of the 13 patients with persistent coagulase-negative staphylococcal bacteremia, 2 received methicillin and 11 received vancomycin. No antibiotic tolerance to either antibiotic could be demonstrated. Serum concentrations of vancomycin far exceeded the minimum bactericidal concentration in all cases in which vancomycin was prescribed. No in vitro differences could be demonstrated between persistent and nonpersistent coagulase-negative staphylococcal strains for slime production, biotype, proteins from modified whole cell lysates developed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and opsonophagocytosis by adult neutrophils in the presence of pooled human sera. Additionally, plasmid profile analysis and phage typing revealed no common strain causing the persistent bacteremia. The pathogenesis of persistent coagulase-negative staphylococcal bacteremia in low birth weight neonates without intravascular catheters remains obscure.

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