scholarly journals Toxicological Characterization and Phospholipase D Activity of the Venom of the Spider Sicarius thomisoides

Toxins ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 702
Author(s):  
Tomás Arán-Sekul ◽  
Ivanka Perčić-Sarmiento ◽  
Verónica Valencia ◽  
Nelly Olivero ◽  
José M. Rojas ◽  
...  
Keyword(s):  
Phospholipase D ◽  
Protein Concentration ◽  
Protein Pattern ◽  
Atacama Desert ◽  
Potential Toxicity ◽  
Cytotoxic Effects ◽  
Dermonecrotic Toxin ◽  
Intraspecific Variations ◽  
Loxosceles Laeta ◽  
Do So

Envenomation by Loxosceles spiders (Sicariidae family) has been thoroughly documented. However, little is known about the potential toxicity of members from the Sicarius genus. Only the venom of the Brazilian Sicarius ornatus spider has been toxicologically characterized. In Chile, the Sicarius thomisoides species is widely distributed in desert and semidesert environments, and it is not considered a dangerous spider for humans. This study aimed to characterize the potential toxicity of the Chilean S. thomisoides spider. To do so, specimens of S. thomisoides were captured in the Atacama Desert, the venom was extracted, and the protein concentration was determined. Additionally, the venoms were analyzed by electrophoresis and Western blotting using anti-recombinant L. laeta PLD1 serum. Phospholipase D enzymatic activity was assessed, and the hemolytic and cytotoxic effects were evaluated and compared with those of the L. laeta venom. The S. thomisoides venom was able to hydrolyze sphingomyelin as well as induce complement-dependent hemolysis and the loss of viability of skin fibroblasts with a dermonecrotic effect of the venom in rabbits. The venom of S. thomisoides showed intraspecific variations, with a similar protein pattern as that of L. laeta venom at 32–35 kDa, recognized by serum anti-LlPLD1. In this context, we can conclude that the venom of Sicarius thomisoides is similar to Loxosceles laeta in many aspects, and the dermonecrotic toxin present in their venom could cause severe harm to humans; thus, precautions are necessary to avoid exposure to their bite.

scholarly journals Novel Mutant Phospholipase D from Hemiscorpius lepturus Acts as A Highly Immunogen in BALB/c Mice Against the Lethality of Scorpion Venom

Molecules ◽  
2020 ◽  
Vol 25 (7) ◽  
pp. 1673 ◽  
Author(s):  
Abouzar Soleimani Moez ◽  
Reza H. Sajedi ◽  
Kamran Pooshang Bagheri ◽  
Jean-Marc Sabatier ◽  
Delavar Shahbazzadeh
Keyword(s):  
Phospholipase D ◽  
Vaccine Candidate ◽  
Clinical Symptoms ◽  
Scorpion Venom ◽  
Effective Vaccine ◽  
Cytotoxic Effects ◽  
In Vivo Tests ◽  
Neurotoxic Effects ◽  
Hemiscorpius Lepturus

Hemiscorpius lepturus (H. lepturus) which belongs to the Scorpionidae family, is the deadliest scorpion in Iran. It causes pathological manifestations like dermonecrosis, hemolysis, renal failure, necrotic ulcers, and in some cases, even death. The venom of this scorpion is well-known for its cytotoxic effects in comparison with the other venomous scorpions which show significant neurotoxic effects. Due to the painless nature of the sting of this scorpion, the clinical symptoms occur in victims 24 to 72 h post-sting. In our previous studies during the last decade, we demonstrated that the medical complications are attributable to the presence of phospholipase D (PLD) as a major toxin in the venom. With the purpose of designing and constructing a vaccine against H. lepturus for humans, animal model experiments were performed. To achieve this goal, non-toxic PLD was developed by mutation of two critical catalytic residues—His12 and His48—into alanines and the product was then denominated mut-rPLD1. The in-vivo tests showed that the mice immunized with interval doses of 10 µg of mut-rPLD1, were completely protected against 10× the LD100 of the venom. In conclusion, this mutant may be an effective vaccine candidate against scorpion envenomation by H. lepturus in future clinical studies.

Tryptophan and aspartic acid residues present in the glycerophosphoryl diester phosphodiesterase (GDPD) domain of the Loxosceles laeta phospholipase D are essential for substrate recognition

Toxicon ◽  
2014 ◽  
Vol 81 ◽  
pp. 43-47 ◽  
Author(s):  
Alejandro Catalán ◽  
William Cortés ◽  
Christian Muñoz ◽  
Jorge E. Araya
Keyword(s):  
Aspartic Acid ◽  
Phospholipase D ◽  
Substrate Recognition ◽  
Loxosceles Laeta

scholarly journals Comparison of two methods of tear sampling for protein quantification by Bradford method

2013 ◽  
Vol 33 (2) ◽  
pp. 261-264 ◽  
Author(s):  
Eliana Farias ◽  
Katia L. Yasunaga ◽  
Romulo V.R. Peixoto ◽  
Micaella P. Fonseca ◽  
Wagner Fontes ◽  
...  
Keyword(s):  
Standard Deviation ◽  
Protein Concentration ◽  
Protein Pattern ◽  
Protein Quantification ◽  
Tear Protein ◽  
Schirmer Tear Test ◽  
Healthy Dogs ◽  
Bradford Method ◽  
Student’S T ◽  
Student’S T Test

The aim of this study was to compare two methods of tear sampling for protein quantification. Tear samples were collected from 29 healthy dogs (58 eyes) using Schirmer tear test (STT) strip and microcapillary tubes. The samples were frozen at -80ºC and analyzed by the Bradford method. Results were analyzed by Student's t test. The average protein concentration and standard deviation from tears collected with microcapillary tube were 4.45mg/mL ±0.35 and 4,52mg/mL ±0.29 for right and left eyes respectively. The average protein concentration and standard deviation from tears collected with Schirmer Tear Test (STT) strip were and 54.5mg/mL ±0.63 and 54.15mg/mL ±0.65 to right and left eyes respectively. Statistically significant differences (p<0.001) were found between the methods. In the conditions in which this study was conducted, the average protein concentration obtained with the Bradford test from tear samples obtained by Schirmer Tear Test (STT) strip showed values higher than those obtained with microcapillary tube. It is important that concentration of tear protein pattern values should be analyzed according the method used to collect tear samples.

Identification of a direct hemolytic effect dependent on the catalytic activity induced by phospholipase-D (dermonecrotic toxin) from brown spider venom

2009 ◽  
Vol 107 (4) ◽  
pp. 655-666 ◽  
Author(s):  
Daniele Chaves-Moreira ◽  
Olga M. Chaim ◽  
Youssef B. Sade ◽  
Kátia S. Paludo ◽  
Luiza H. Gremski ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Phospholipase D ◽  
Spider Venom ◽  
Dermonecrotic Toxin ◽  
Hemolytic Effect ◽  
Brown Spider

scholarly journals Phospholipase D from Loxosceles laeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration

Toxins ◽  
2017 ◽  
Vol 9 (4) ◽  
pp. 125 ◽  
Author(s):  
José Rojas ◽  
Tomás Arán-Sekul ◽  
Emmanuel Cortés ◽  
Romina Jaldín ◽  
Kely Ordenes ◽  
...  
Keyword(s):  
Human Skin ◽  
Phospholipase D ◽  
Skin Fibroblasts ◽  
Spider Venom ◽  
Human Skin Fibroblasts ◽  
Loxosceles Laeta

Kinetics of cell spreading in the presence of different concentrations of serum or fibronectin-depleted serum

1984 ◽  
Vol 71 (1) ◽  
pp. 51-59
Author(s):  
P. Knox
Keyword(s):  
Serum Concentration ◽  
Plasma Proteins ◽  
Protein Concentration ◽  
Inverse Relationship ◽  
Cell Spreading ◽  
Serum Concentrations ◽  
Spreading Factor ◽  
Exogenous Protein ◽  
Kinetics Of ◽  
Do So

A morphometric assay has been used to show that speed of cell spreading is sensitive to the level of serum in the surrounding growth medium. There was an inverse relationship between serum concentration and speed of spreading; as serum concentration was increased from 0.1% to 100% cells took progressively longer to become fully spread. The time taken to achieve complete spreading in serum concentrations of 0.1, 1.0, 10 and 100% serum was 45, 60, 105 and 240 min, respectively. At serum concentrations of 3% and above, fibronectin depletion had no effect on rates of spreading. Only at concentrations below 3% was there an effect of fibronectin depletion and although cells became fully spread in the depleted serum they took longer to do so in comparison to native serum. It is suggested that plasma fibronectin plays no role in serum-stimulated spreading at serum concentrations of 3% and above since at these concentrations rates of spreading are identical whether fibronectin is present or not. At concentrations of 3% and above spreading is mediated by the 70K spreading factor. At serum concentrations below 3% both fibronectin and 70K factor are effective. The reason for the lack of effect of fibronectin at higher serum concentrations is an inhibition by albumin and other plasma proteins. In contrast, 70K factor is not affected by exogenous protein concentration. The effect of exogenous protein concentration on cell adhesion is discussed in terms of cell migration between compartments that have different extracellular protein concentrations.

Identification, cloning and functional characterization of a novel dermonecrotic toxin (phospholipase D) from brown spider (Loxosceles intermedia) venom

2008 ◽  
Vol 1780 (2) ◽  
pp. 167-178 ◽  
Author(s):  
Marcia Helena Appel ◽  
Rafael Bertoni da Silveira ◽  
Olga Meiri Chaim ◽  
Kátia Sabrina Paludo ◽  
Dilza Trevisan Silva ◽  
...  
Keyword(s):  
Phospholipase D ◽  
Functional Characterization ◽  
Dermonecrotic Toxin ◽  
Brown Spider

Coupling of p-nitropnenyl diazonium, fluoroborate to bovine chymotrypsinogen A and α-chymotrypsin

1969 ◽  
Vol 47 (4) ◽  
pp. 423-427 ◽  
Author(s):  
V. T. Maddaiah
Keyword(s):  
Protein Concentration ◽  
Model Compound ◽  
Caproic Acid ◽  
Acetate Buffer ◽  
Reaction Products ◽  
Amino Groups ◽  
Native Proteins ◽  
Equal Rate ◽  
Spectral Comparison ◽  
Do So

p-Nitrophenyl diazonium fluoroborate couples at the ε-amino groups of lysyl residues of bovine chymotrypsinogen A and α-chymotrypsin in 0.5 M acetate buffer, pH 5.0. The reaction products are identified by spectral comparison with the model compound bis-(p-nitrophenylazo)-ε-amino-n-caproic acid. The reaction with the zymogen is about four times faster than with the active enzyme and this difference is independent of protein concentration. However, in the presence of 8 M urea, the reaction occurs at the histidyl groups at an equal rate. This suggests that of the groups that could undergo coupling, i.e. histidyl, tyrosyl, and lysyl, only the latter do so in the native proteins; moreover, chymotrypsinogen appears to have more such lysyl residues than does chymotrypsin.

The Correspondence with Beer's Law for the Optical Density of Stained Protein Patterns on Filter Paper as a Function of Surface Protein Concentration

1955 ◽  
Vol 1 (5) ◽  
pp. 329-344 ◽  
Author(s):  
V H Rees ◽  
D J R Laurence
Keyword(s):  
Protein Concentration ◽  
Protein Pattern ◽  
Surface Protein ◽  
General Situation ◽  
Protein Patterns ◽  
Experimental Conditions ◽  
Scanning Method ◽  
Beer's Law ◽  
Beer’S Law ◽  
Protein Density

Abstract The results obtained here consistently failed to show deviations from Beer's law for optical densities less than 1.4, and the use of scanning as a convenient method for differential protein estimations would appear to be justified. The methods described may enable other workers to make similar tests of the method with a minimum of preliminary development, and some may succeed in obtaining significant failure of Beer's law in their apparatus. The use of apparatus of this type would enable the variation of the deviations with arrangement of the optical system, etc., to be worked out, but the following experimental conditions are already known to lead to significant deviations: 1. Use of dry paper instead of lightly oiled paper. 2. An inadequate light ifiter for the photocell. 3. An illuminated slit too long to be completely covered by the protein pattern. 4. Variations in protein density over the slit, either because the slit is too wide or because the protein pattern has been applied unevenly. 5. Use of low quality ifiter paper containing "pin holes." 6. A dye uptake which is not proportional to protein content of the paper [Martin and Franglen (14)]. The failure of Crook, Harris, and Warren (4) to substantiate Beer's law does not indicate the most general situation for the application of the scanning method. The application of Beer's logarithmic law to the scanning of dyed protein patterns has been investigated by methods described in detail. No deviations could be found for optical densities less than 1.4 for amidoschwarz 1OB or less than 1.1 for azocarmine B staining. The scanning method can be used for evaluation of protein fractions if care is taken.

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