Biochemical and pharmacological characterization of PhTX-I a new myotoxic phospholipase A(2) isolated from Porthidium hyoprora snake venom

AbstractThis paper reports the biochemical and pharmacological characterization of a new myotoxic PLA2 (EC 3.1.1.4) called PhTX-I, purified from Porthidium hyoprora venom by one step analytical chromatography reverse phase HPLC. The homogeneity of the PhTX-I fraction and its molecular mass were initially evaluated by SDS–PAGE and confirmed by MALDI-TOF spectrometry, indicating a molecular mass of 14.249Da and constituted of a single polipeptidic chain. Amino acid sequence was determined by “de novo sequencing,” in tandem mass spectrometry, belonging to D49-PLA2 enzyme class and exhibiting high identity (44–90%) with other myotoxics PLA2 from snake venoms. The enzymatic investigation showed maximal activity at pH 8 and 35–45°C. This activity was dependent on Ca2+, other cations (Mg2+, Mn2+, Cd2+ and Zn2+) reduced notably the enzymatic activity, suggesting that the arrangement of the catalytic site presents an exclusive structure for Ca2+. Ex vivo, whole venom and PhTX-I PLA2 caused blockade of the neuromuscular transmission in young chick biventer cervicis preparations similar to other isolated snake venom toxins from the Bothrops genus. In vivo, both induced local myotoxicity and systemic interleukin-6 response upon intramuscular injection, additionally, induced moderate footpad edema. In vitro, both induced low cytotoxicity in skeletal muscle myoblasts, however PhTX-I PLA2 was able to lyse myotubes

Concern About Returning to Face-to-Face Classes After the Pandemic: Importance of Emotional Intelligence and Stress Coping Strategies in Health Science Students

Mayela Cajachagua Castro,1 Keila Miranda Limachi,1 Janett V Chávez Sosa,1 Salomon Huancahuire-Vega2 1Nursing Department, School of Health Sciences, Universidad Peruana Unión (UPeU), Lima, Peru; 2Basic Sciences Department, Human Medicine School, Universidad Peruana Unión (UPeU), Lima, PeruCorrespondence: Salomon Huancahuire-Vega, Basic Sciences Department, Human Medicine School, Universidad Peruana Unión (UPeU), Carretera Central Km 19, Ñaña (Prolong. Bernardo Balaguer), Lurigancho, Lima, 15464, Peru, Tel +51997574011, Email [email protected]: The COVID-19 pandemic has brought a new normality, a scenario where different circumstances trigger emotions such as concern about returning to face-to-face classes, to which higher education students adapt. The objective was to identify how emotional intelligence and coping with stress explain students’ concern about returning to face-to-face classes post COVID-19.Methods: This was an explanatory and cross-sectional study. The sample by intention was 588 students from the Faculty of Health Sciences. The Rotterdam Emotional Intelligence Scale (REIS), the Stress Coping Questionnaire (CAE), and the ACAD-COVID-19 scale were used. For data collection, the instruments were digitized. Bivariate analysis with Chi-square and multivariate logistic regression was performed.Results: The mean age of the participants was 19.72 years; most were single and without children (96.9% and 96.8%, respectively). 74.3% did not work, 80.8% were from the first years of study, and 52.7% belonged to nursing school. About 94.2% of participants indicated having adequate emotional intelligence, 91.3% expressed coping with adequate stress, and 67.2% indicated serious concern about returning to face-to-face classes. An association was found between gender (p=0.042), age (p=0.002), year of study (p=0.027), emotional intelligence (p=0.001), and coping with stress (p=0.001) with concern for return to face-to-face classes. Emotional intelligence identified as adequate (OR: 2.580; IC95%: 1.117– 5.960) and coping with stress identified as adequate (OR: 2.008; IC95%: 1.018– 3.960) are more likely to express serious concern about the return to face-to-face classes after the COVID-19 pandemic.Conclusion: According to the results, the need to safeguard the psychological aspects of students is highlighted, especially emotional intelligence, as well as to improve coping with stress so that they can better manage concerns about returning to face-to-face classes. The educational authorities should implement strategies to improve these aspects in order to ensure the adequate return to face-to-face classes in new scenarios.Keywords: emotional intelligence, coping, psychological stress, expression of concern, health science student

cDNA and deduced primary structure of basic phospholipase A2 with neurotoxic activity from the venom secretion of the Crotalus durissus collilineatus rattlesnake

To illustrate the construction of precursor complementary DNAs, we isolated mRNAs from whole venom samples. After reverse transcription polymerase chain reaction (RT-PCR), we amplified the cDNA coding for a neurotoxic protein, phospholipase A2 D49 (PLA2 D49), from the venom of Crotalus durissus collilineatus (Cdc PLA2). The cDNA encoding Cdc PLA2 from whole venom was sequenced. The deduced amino acid sequence of this cDNA has high overall sequence identity with the group II PLA2 protein family. Cdc PLA2 has 14 cysteine residues capable of forming seven disulfide bonds that characterize this group of PLA2 enzymes. Cdc PLA2 was isolated using conventional Sephadex G75 column chromatography and reverse-phase high performance liquid chromatography (RP-HPLC). The molecular mass was estimated using matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry. We tested the neuromuscular blocking activities on chick biventer cervicis neuromuscular tissue. Phylogenetic analysis of Cdc PLA2 showed the existence of two lines of N6-PLA2, denominated F24 and S24. Apparently, the sequences of the New World’s N6-F24-PLA2 are similar to those of the agkistrodotoxin from the Asian genus Gloydius. The sequences of N6-S24-PLA2 are similar to the sequence of trimucrotoxin from the genus Protobothrops, found in the Old World

Chemical Modifications Of Phtx-i Myotoxin From Porthidium Hyoprora Snake Venom: Effects On Structural, Enzymatic, And Pharmacological Properties

We recently described the isolation of a basic PLA2 (PhTX-I) from Porthidium hyoprora snake venom. This toxin exhibits high catalytic activity, induces in vivo myotoxicity, moderates footpad edema, and causes in vitro neuromuscular blockade. Here, we describe the chemical modifications of specific amino acid residues (His, Tyr, Lys, and Trp), performed in PhTX-I, to study their effects on the structural, enzymatic, and pharmacological properties of this myotoxin. After chemical treatment, a single His, 4 Tyr, 7 Lys, and one Trp residues were modified. The secondary structure of the protein remained unchanged as measured by circular dichroism; however other results indicated the critical role played by Lys and Tyr residues in myotoxic, neurotoxic activities and mainly in the cytotoxicity displayed by PhTX-I. His residue and therefore catalytic activity of PhTX-I are relevant for edematogenic, neurotoxic, and myotoxic effects, but not for its cytotoxic activity. This dissociation observed between enzymatic activity and some pharmacological effects suggests that other molecular regions distinct from the catalytic site may also play a role in the toxic activities exerted by this myotoxin. Our observations supported the hypothesis that both the catalytic sites as the hypothetical pharmacological sites are relevant to the pharmacological profile of PhTX-I. © 2013 Salomón Huancahuire-Vega et al.2013Kini, R.M., Excitement ahead: Structure, function and mechanism of snake venom phospholipase A2 enzymes (2003) Toxicon, 42 (8), pp. 827-840. , DOI 10.1016/j.toxicon.2003.11.002Pungercar, J., Krizaj, I., Understanding the molecular mechanism underlying the presynaptic toxicity of secreted phospholipases A2 (2007) Toxicon, 50 (7), pp. 871-892. , DOI 10.1016/j.toxicon.2007.07.025, PII S0041010107002875Gutiérrez, J.M., Ponce-Soto, L.A., Marangoni, S., Lomonte, B., Systemic and local myotoxicity induced by snake venom group II phospholipases A2: Comparison between crotoxin, crotoxin B and a Lys49 PLA2 homologue (2008) Toxicon, 51 (1), pp. 80-92. , 2-s2.0-38049077979 10.1016/j.toxicon.2007.08.007Warrell, D.A., Snake bite (2010) The Lancet, 375 (9708), pp. 77-88. , 2-s2.0-73049107213 10.1016/S0140-6736(09)61754-2Huang, M.Z., Wang, Q.C., Liu, G.F., Effects of an acidic phospholipase A2 purified from Ophiophagus hannah (king cobra) venom on rat heart (1993) Toxicon, 31 (5), pp. 627-635. , 2-s2.0-0027160339 10.1016/0041-0101(93)90117-2Atanasov, V.N., Danchev, D., Mitewa, M., Petrova, S., Hemolytic and anticoagulant study of the neurotoxin vipoxin and its components-basic phospholipase A2 and an acidic inhibitor (2009) Biochemistry, 74 (3), pp. 276-280. , 2-s2.0-65549140342 10.1134/S0006297909030055Rudrammaji, L.M.S., Machiah, K.D., Kantha, T.P.K., Gowda, T.V., Role of catalytic function in the antiplatelet activity of phospholipase A2 cobra (Naja naja naja) venom (2001) Molecular and Cellular Biochemistry, 219 (1-2), pp. 39-44. , DOI 10.1023/A:1011002606505Cicala, C., Cirino, G., Phospholipase A2-induced hypotension in the rat and its pharmacological modulation (1993) General Pharmacology, 24 (5), pp. 1197-1202Francis, B.R., Jorge Da Silva Jr., N., Seebart, C., Casais E Silva, L.L., Schmidt, J.J., Kaiser, I.I., Toxins isolated from the venom of the Brazilian coral snake (Micrurus frontalis frontalis) include hemorrhagic type phospholipases A2 and postsynaptic neurotoxins (1997) Toxicon, 35 (8), pp. 1193-1203. , DOI 10.1016/S0041-0101(97)00031-7, PII S0041010197000317Ferreira, T., Camargo, E.A., Ribela, M.T.C.P., Damico, D.C., Marangoni, S., Antunes, E., De Nucci, G., Landucci, E.C.T., Inflammatory oedema induced by Lachesis muta muta (Surucucu) venom and LmTX-I in the rat paw and dorsal skin (2009) Toxicon, 53 (1), pp. 69-77. , 2-s2.0-57749111830 10.1016/j.toxicon.2008.10.016Ponce-Soto, L.A., Lomonte, B., Gutierrez, J.M., Rodrigues-Simioni, L., Novello, J.C., Marangoni, S., Structural and functional properties of BaTX, a new Lys49 phospholipase A2 homologue isolated from the venom of the snake Bothrops alternatus (2007) Biochimica et Biophysica Acta - General Subjects, 1770 (4), pp. 585-593. , DOI 10.1016/j.bbagen.2006.11.015, PII S0304416506003655Ponce-Soto, L.A., Martins-De-Souza, D., Marangoni, S., Neurotoxic, myotoxic and cytolytic activities of the new basic PLA 2 isoforms BmjeTX-I and BmjeTX-II isolated from the Bothrops marajoensis (marajó lancehead) snake venom (2010) Protein Journal, 29 (2), pp. 103-113. , 2-s2.0-77953250412 10.1007/s10930-010-9229-5Arni, R.K., Ward, R.J., Phospholipase A2 - A structural review (1996) Toxicon, 34 (8), pp. 827-841. , DOI 10.1016/0041-0101(96)00036-0Petan, T., Krizaj, I., Pungercar, J., Restoration of enzymatic activity in a Ser-49 phospholipase A2 homologue decreases its Ca2+-independent membrane-damaging activity and increases its toxicity (2007) Biochemistry, 46 (44), pp. 12795-12809. , DOI 10.1021/bi701304eLomonte, B., Angulo, Y., Sasa, M., Gutiérrez, J.M., The phospholipase A2 homologues of snake venoms: Biological activities and their possible adaptive roles (2009) Protein and Peptide Letters, 16 (8), pp. 860-876. , 2-s2.0-68849126695 10.2174/092986609788923356Huancahuire-Vega, S., Ponce-Soto, L.A., Martins-De-Souza, D., Marangoni, S., Structural and functional characterization of brazilitoxins II and III (BbTX-II and -III), two myotoxins from the venom of Bothrops brazili snake (2009) Toxicon, 54 (6), pp. 818-827. , 2-s2.0-68949148773 10.1016/j.toxicon.2009.06.008Jensen, L.B., Burgess, N.K., Gonda, D.D., Spencer, E., Wilson-Ashworth, H.A., Driscoll, E., Vu, M.P., Bell, J.D., Mechanisms governing the level of susceptibility of erythrocyte membranes to secretory phospholipase A2 (2005) Biophysical Journal, 88 (4), pp. 2692-2705. , DOI 10.1529/biophysj.104.056457Jenkins, C.M., Cedars, A., Gross, R.W., Eicosanoid signalling pathways in the heart (2009) Cardiovascular Research, 82 (2), pp. 240-249. , 2-s2.0-67449132320 10.1093/cvr/cvn346Cintra-Francischinelli, M., Pizzo, P., Angulo, Y., Gutiérrez, J.M., Montecucco, C., Lomonte, B., The C-terminal region of a Lys49 myotoxin mediates CA2+ influx in C2C12 myotubes (2010) Toxicon, 55 (2-3), pp. 590-596. , 2-s2.0-72749095520 10.1016/j.toxicon.2009.10.013Huancahuire-Vega, S., Ponce-Soto, L.A., Martins-De-Souza, D., Marangoni, S., Biochemical and pharmacological characterization of PhTX-I a new myotoxic phospholipase A2 isolated from Porthidium hyoprora snake venom (2011) Comparative Biochemistry and Physiology C, 29, pp. 103-113. , 2-s2.0-79955099265 10.1016/j.cbpc.2011.03.013Diaz-Oreiro, C., Gutierrez, J.M., Chemical modification of histidine and lysine residues of myotoxic phospholipases A2 isolated from Bothrops asper and Bothrops godmani snake venoms: Effects on enzymatic and pharmacological properties (1997) Toxicon, 35 (2), pp. 241-252. , DOI 10.1016/S0041-0101(96)00128-6, PII S0041010196001286Soares, A.M., Andriao-Escarso, S.H., Bortoleto, R.K., Rodrigues-Simioni, L., Arni, R.K., Ward, R.J., Gutierrez, J.M., Giglio, J.R., Dissociation of enzymatic and pharmacological properties of piratoxins-I and -III, two myotoxic phospholipases A2 from Bothrops pirajai snake venom (2001) Archives of Biochemistry and Biophysics, 387 (2), pp. 188-196. , DOI 10.1006/abbi.2000.2244Soares, A.M., Giglio, J.R., Chemical modifications of phospholipases A2 from snake venoms: Effects on catalytic and pharmacological properties (2003) Toxicon, 42 (8), pp. 855-868. , DOI 10.1016/j.toxicon.2003.11.004Soons, K.R., Condrea, E., Yang, C.-C., Rosenberg, P., Effects of modification of tyrosines 3 and 62 (63) on enzymatic and toxicological properties of phospholipases A2 from Naja nigricollis and Naja naja atra snake venoms (1986) Toxicon, 24 (7), pp. 679-693. , DOI 10.1016/0041-0101(86)90031-0Takasaki, C., Sugama, A., Yanagita, A., Tamiya, N., Rowan, E.G., Harvey, A.L., Effects of chemical modifications of Pa-11, a phospholipase A2 from the venom of Australian king brown snake (Pseudechis australis), on its biological activities (1990) Toxicon, 28 (1), pp. 107-117. , DOI 10.1016/0041-0101(90)90012-VHeinrikson, R.L., Meredith, S.C., Amino acid analysis by reverse-phase high-performance liquid chromatography: Precolumn derivatization with phenylisothiocyanate (1984) Analytical Biochemistry, 136 (1), pp. 65-74. , 2-s2.0-0021312893Schagger, H., Von Jagow, G., Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa (1987) Analytical Biochemistry, 166 (2), pp. 368-379. , DOI 10.1016/0003-2697(87)90587-2Corrêa, D.H.A., Ramos, C.H.I., The use of circular dichroism spectroscopy to study protein folding, form and function (2009) African Journal of Biochemistry Research, 3, pp. 164-173Cho, W., Kezdy, F.J., Chromogenic substrates and assay of phospholipases A2 (1991) Methods in Enzymology, 197, pp. 75-79. , 2-s2.0-0025822798Holzer, M., Mackessy, S.P., An aqueous endpoint assay of snake venom phospholipase A2 (1996) Toxicon, 34 (10), pp. 1149-1155. , DOI 10.1016/0041-0101(96)00057-8, PII S0041010196000578Borenfreund, E., Borrero, O., In vitro cytotoxicity assays. Potential alternatives to the Draize ocular allergy test (1984) Cell Biology and Toxicology, 1 (1), pp. 55-65. , DOI 10.1007/BF00125565Yang, C.C., Chang, L.S., Dissociation of lethal toxicity and enzymic activity of notexin from Notechis scutatus scutatus (Australian-tiger-snake) venom by modification of tyrosine residues (1991) Biochemical Journal, 280 (3), pp. 739-744. , 2-s2.0-0026335259Soares, A.M., Guerra-Sa, R., Borja-Oliveira, C.R., Rodrigues, V.M., Rodrigues-Simioni, L., Rodrigues, V., Fontes, M.R.M., Giglio, J.R., Structural and functional characterization of BnSP-7, a Lys49 myotoxic phospholipase A2 homologue from Bothrops neuwiedi pauloensis venom (2000) Archives of Biochemistry and Biophysics, 378 (2), pp. 201-209. , DOI 10.1006/abbi.2000.1790Rosenburg, P., Ghassemi, A., Condrea, E., Dhillon, D., Yang, C.-C., Do chemical modifications dissociate between the enzymatic and pharmacological activities of β bungarotoxin and notexin? (1989) Toxicon, 27 (2), pp. 137-159. , DOI 10.1016/0041-0101(89)90128-1Andrião-Escarso, S.H., Soares, A.M., Rodrigues, V.M., Angulo, Y., Díaz, C., Lomonte, B., Gutiérrez, J.M., Giglio, J.R., Myotoxic phospholipases A2 in Bothrops snake venoms: Effect of chemical modifications on the enzymatic and pharmacological properties of bothropstoxins from Bothrops jararacussu (2000) Biochimie, 82 (8), pp. 755-763. , 2-s2.0-0033819610 10.1016/S0300-9084(00)01150-0Bazaa, A., Luis, J., Srairi-Abid, N., Kallech-Ziri, O., Kessentini-Zouari, R., Defilles, C., Lissitzky, J.C., Marrakchi, N., MVL-PLA2, a phospholipase A2 from Macrovipera lebetina transmediterranea venom, inhibits tumor cells adhesion and migration (2009) Matrix Biology, 28 (4), pp. 188-193. , 2-s2.0-67349183397 10.1016/j.matbio.2009.03.007Scott, D.L., Achari, A., Vidal, J.C., Sigler, P.B., Crystallographic and biochemical studies of the (inactive) Lys-49 phospholipase A2 from the venom of Agkistridon piscivorus piscivorus (1992) Journal of Biological Chemistry, 267 (31), pp. 22645-22657. , 2-s2.0-0026499354Kao, P.-H., Chen, K.-C., Lin, S.-R., Chang, L.-S., The structural and functional contribution of N-terminal region and His-47 on Taiwan cobra phospholipase A2 (2008) Journal of Peptide Science, 14 (3), pp. 342-348. , DOI 10.1002/psc.943Diz Filho, E.B.S., Marangoni, S., Toyama, D.O., Fagundes, F.H.R., Oliveira, S.C.B., Fonseca, F.V., Calgarotto, A.K., Toyama, M.H., Enzymatic and structural characterization of new PLA2 isoform isolated from white venom of Crotalus durissus ruruima (2009) Toxicon, 53 (1), pp. 104-114. , 2-s2.0-57849150819 10.1016/j.toxicon.2008.10.021Toyama, D.D.O., Diz Filho, E.B.D.S., Cavada, B.S., Da Rocha, B.A.M., De Oliveira, S.C.B., Cotrim, C.A., Soares, V.C.G., Toyama, M.H., Umbelliferone induces changes in the structure and pharmacological activities of Bn IV, a phospholipase A2 isoform isolated from Bothrops neuwiedi (2011) Toxicon, 57 (6), pp. 851-860. , 2-s2.0-79954995046 10.1016/j.toxicon.2011.02.024Beghini, D.G., Toyama, M.H., Hyslop, S., Sodek, L.C., Novello, Marangoni, S., Enzymatic characterization of a novel phospholipase A2 from Crotalus durissus cascavella rattlesnake (maracambóia) venom (2000) Journal of Protein Chemistry, 19 (8), pp. 679-684. , DOI 10.1023/A:1007152303179Shina, R., Yates, S.L., Ghassemi, A., Rosenberg, P., Condrea, E., Inhibitory effect of EDTA·Ca2+ on the hydrolysis of synaptosomal phospholipids by phospholipase A2 toxins and enzymes (1990) Biochemical Pharmacology, 40 (10), pp. 2233-2239. , DOI 10.1016/0006-2952(90)90717-YBonfim, V.L., Toyama, M.H., Novello, J.C., Hyslop, S., Oliveira, C.R.B., Rodrigues-Simioni, L., Marangoni, S., Isolation and enzymatic characterization of a basic phospholipase A 2 from Bothrops jararacussu snake venom (2001) Protein Journal, 20 (3), pp. 239-245. , 2-s2.0-52549108470Yang, C.C., Kini, R.M., Chemical modification and functional sites of phospholipases A 2 (1997) Venom Phospholipase A2 Enzymes: Structure, Function and Mechanism, pp. 185-204. , Chichester, UK WileyRangel, J., Quesada, O., Gutiérrez, J.M., Angulo, Y., Lomonte, B., Membrane cholesterol modulates the cytolytic mechanism of myotoxin II, a Lys49 phospholipase A2 homologue from the venom of Bothrops asper (2011) Cell Biochemistry and Function, 29 (5), pp. 365-370. , 2-s2.0-79960082194 10.1002/cbf.1758Lomonte, B., Angulo, Y., Calderon, L., An overview of lysine-49 phospholipase A2 myotoxins from crotalid snake venoms and their structural determinants of myotoxic action (2003) Toxicon, 42 (8), pp. 885-901. , DOI 10.1016/j.toxicon.2003.11.008Angulo, Y., Lomonte, B., Differential susceptibility of C2C12 myoblasts and myotubes to group II phospholipase A2 myotoxins from crotalid snake venoms (2005) Cell Biochemistry and Function, 23 (5), pp. 307-313. , DOI 10.1002/cbf.1208Gebrim, L.C., Marcussi, S., Menaldo, D.L., De Menezes, C.S.R., Nomizo, A., Hamaguchi, A., Silveira-Lacerda, E.P., Rodrigues, V.M., Antitumor effects of snake venom chemically modified Lys49 phospholipase A2-like BthTX-I and a synthetic peptide derived from its C-terminal region (2009) Biologicals, 37 (4), pp. 222-229. , 2-s2.0-67650751847 10.1016/j.biologicals.2009.01.010Romero-Vargas, F.F., Ponce-Soto, L.A., Martins-De-Souza, D., Marangoni, S., Biological and biochemical characterization of two new PLA2 isoforms Cdc-9 and Cdc-10 from Crotalus durissus cumanensis snake venom (2010) Comparative Biochemistry and Physiology C, 151 (1), pp. 66-74. , 2-s2.0-71649092384 10.1016/j.cbpc.2009.08.011Costa, T.R., Menaldo, D.L., Oliveira, C.Z., Santos-Filho, N.A., Teixeira, S.S., Nomizo, A., Fuly, A.L., Soares, A.M., Myotoxic phospholipases A2 isolated from Bothrops brazili snake venom and synthetic peptides derived from their C-terminal region: Cytotoxic effect on microorganism and tumor cells (2008) Peptides, 29 (10), pp. 1645-1656. , 2-s2.0-52249108502 10.1016/j.peptides.2008.05.021Cummings, J., Hodgkinson, C., Odedra, R., Sini, P., Heaton, S.P., Mundt, K.E., Ward, T.H., Dive, C., Preclinical evaluation of M30 and M65 ELISAs as biomarkers of drug induced tumor cell death and antitumor activity (2008) Molecular Cancer Therapeutics, 7 (3), pp. 455-463. , http://mct.aacrjournals.org/cgi/reprint/7/3/455, DOI 10.1158/1535-7163.MCT-07-2136Panini, S.R., Yang, L., Rusinol, A.E., Sinensky, M.S., Bonventre, J.V., Leslie, C.C., Arachidonate metabolism and the signaling pathway of induction of apoptosis by oxidized LDL/oxysterol (2001) Journal of Lipid Research, 42 (10), pp. 1678-1686Teixeira, C.F.P., Landucci, E.C.T., Antunes, E., Chacur, M., Cury, Y., Inflammatory effects of snake venom myotoxic phospholipases A2 (2003) Toxicon, 42 (8), pp. 947-962. , DOI 10.1016/j.toxicon.2003.11.006Teixeira, C., Cury, Y., Moreira, V., Picolo, G., Chaves, F., Inflammation induced by Bothrops asper venom (2009) Toxicon, 54 (7), pp. 988-997. , 2-s2.0-70149099543 10.1016/j.toxicon.2009.05.026Zuliani, J.P., Fernandes, C.M., Zamuner, S.R., Gutierrez, J.M., Teixeira, C.F.P., Inflammatory events induced by Lys-49 and Asp-49 phospholipases A 2 isolated from Bothrops asper snake venom: Role of catalytic activity (2005) Toxicon, 45 (3), pp. 335-346. , DOI 10.1016/j.toxicon.2004.11.004Vishwanath, B.S., Kini, R.M., Gowda, T.V., Characterization of three edema-inducing phospholipase A2 enzymes from habu (Trimeresurus flavoviridis) venom and their interaction with the alkaloid aristolochic acid (1987) Toxicon, 25 (5), pp. 501-509. , 2-s2.0-0023186213Soares, A.M., Sestito, W.P., Marcussi, S., Stabeli, R.G., Andriao-Escarso, S.H., Cunha, O.A.B., Vieira, C.A., Giglio, J.R., Alkylation of myotoxic phospholipases A2 in Bothrops moojeni venom: A promising approach to an enhanced antivenom production (2004) International Journal of Biochemistry and Cell Biology, 36 (2), pp. 258-270. , DOI 10.1016/S1357-2725(03)00237-1Zhao, H., Tang, L., Wang, X., Zhou, Y., Zhengjiong Lin, Structure of a snake venom phospholipase A2 modified by p-bromo- phenacyl-bromide (1998) Toxicon, 36 (6), pp. 875-886. , DOI 10.1016/S0041-0101(97)00169-4, PII S0041010197001694Araya, C., Lomonte, B., Antitumor effects of cationic synthetic peptides derived from Lys49 phospholipase A2 homologues of snake venoms (2007) Cell Biology International, 31 (3), pp. 263-268. , DOI 10.1016/j.cellbi.2006.11.007, PII S1065699506002691Stábeli, R.G., Amui, S.F., Sant'Ana, C.D., Pires, M.G., Nomizo, A., Monteiro, M.C., Romão, P.R.T., Soares, A.M., Bothrops moojeni myotoxin-II, a Lys49-phospholipase A2 homologue: An example of function versatility of snake venom proteins (2006) Comparative Biochemistry and Physiology C, 142 (3-4), pp. 371-381. , 2-s2.0-33644993571 10.1016/j.cbpc.2005.11.02

Association Between Family Support and Coping Strategies of People With Covid-19: A Cross-Sectional Study

Aileen M Chilon-Huaman,1 Ángela Camposano-Ninahuanca,1 Janett V Chávez-Sosa,1 Salomon Huancahuire-Vega,2,3 William De Borba1 1Nursing School, Universidad Peruana Unión (UPeU), Lima, Peru; 2Human Medicine School, Universidad Peruana Unión (UPeU), Lima, Peru; 3General Directorate of Research, Universidad Peruana Unión, (UPeU), Lima, PeruCorrespondence: Salomon Huancahuire-Vega, Human Medicine School, Universidad Peruana Unión (UPeU), Carretera Central Km 19 Lurigancho-Chosica, Lima, Peru, Tel +51 997 574 011, Email [email protected]: The study aimed to determine the association between family support and coping strategies of people diagnosed with COVID-19.Methods: The study was analytical and cross-sectional. The sample consisted of 500 participants who were selected by non-probabilistic and snowball sampling and included residents of both sexes who belonged to the city of Lima, with a diagnosis of COVID-19, who lived with relatives, and who accepted to participate in the research. For data collection, the scales “family support” and “Coping and Adaptation Process-Coping Adaptation Processing Scale (CAPS)” were used. The technique used was the survey through the home visit and the questionnaire instrument. To measure the relationship of the study variables, binary logistic regression was chosen, considering coping strategies as the dependent variable and socio-demographic data and family support as independent variables.Results: Of the 500 participants, 50.4% were women, and 49.6% were men. The results revealed that most participants presented a high capacity for coping strategies and high perceived family support (97.2% and 81%, respectively). In the bivariate analysis, socio-demographic aspects and family support and their dimensions were related to high or low capacity for coping strategies. Significant differences were verified between marital status (p=0.026), having children (p=0.037), family support (p=0.000), and its dimensions with coping strategies. Finally, the multivariate analysis found that people with COVID-19 who perceived high family support were 33.74 times (95% CI: 7266– 156,739) more likely to have a high capacity for coping strategies.Conclusion: Therefore, it is necessary to promote the development of parental and family support skills in the face of the health emergency caused by COVID-19.Keywords: family support, coping strategies, coronavirus, health emergenc

Chemical Modi cations of PhTX-I Myoto in from Porthidium hyoprora Snake Venom: Effects on Structural, Enzymatic, and Pharmacological Properties

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)We recently described the isolation of a basic PLA2 (PhTX-I) from Porthidium hyoprora snake venom.. is toxin exhibits high catalytic activity, induces in vivo myotoxicity, moderates footpad edema, and causes in vitro neuromuscular blockade. Here, we describe the chemical modifications of specific amino acid residues (His, Tyr, Lys, and Trp), performed in PhTX-I, to study their effects on the structural, enzymatic, and pharmacological properties of this myotoxin. After chemical treatment, a single His, 4 Tyr, 7 Lys, and one Trp residues were modified. The secondary structure of the protein remained unchanged as measured by circular dichroism; however other results indicated the critical role played by Lys and Tyr residues in myotoxic, neurotoxic activities and mainly in the cytotoxicity displayed by PhTX-I. His residue and therefore catalytic activity of PhTX-I are relevant for edematogenic, neurotoxic, and myotoxic effects, but not for its cytotoxic activity. This dissociation observed between enzymatic activity and some pharmacological effects suggests that other molecular regions distinct from the catalytic site may also play a role in the toxic activities exerted by this myotoxin. Our observations supported the hypothesis that both the catalytic sites as the hypothetical pharmacological sites are relevant to the pharmacological profile of PhTX-I.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Mass spectrometry Laboratory at Brazilian Biosciences National laboratory, CNPEM-ABTLUS, Campinas, BrazilFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP [09/51207-9

Ejendomsforholdene i Hindsted Herred i Slutningen af Middelalderen.

To illustrate the construction of precursor complementary DNAs, we isolated mRNAs from whole venom samples. After reverse transcription polymerase chain reaction (RT-PCR), we amplified the cDNA coding for a neurotoxic protein, phospholipase A2 D49 (PLA2 D49), from the venom of Crotalus durissus collilineatus (Cdc PLA2). The cDNA encoding Cdc PLA2 from whole venom was sequenced. The deduced amino acid sequence of this cDNA has high overall sequence identity with the group II PLA2 protein family. Cdc PLA2 has 14 cysteine residues capable of forming seven disulfide bonds that characterize this group of PLA2 enzymes. Cdc PLA2 was isolated using conventional Sephadex G75 column chromatography and reverse-phase high performance liquid chromatography (RP-HPLC). The molecular mass was estimated using matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry. We tested the neuromuscular blocking activities on chick biventer cervicis neuromuscular tissue. Phylogenetic analysis of Cdc PLA2 showed the existence of two lines of N6-PLA2, denominated F24 and S24. Apparently, the sequences of the New World's N6-F24-PLA2 are similar to those of the agkistrodotoxin from the Asian genus Gloydius. The sequences of N6-S24-PLA2 are similar to the sequence of trimucrotoxin from the genus Protobothrops, found in the Old World.43262-7