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1.
J Proteome Res ; 23(8): 3638-3648, 2024 Aug 02.
Artículo en Inglés | MEDLINE | ID: mdl-39038168

RESUMEN

Heloderma horridum horridum, a venomous reptile native to America, has a venom with potential applications in treating type II diabetes. In this work, H. h. horridum venom was extracted, lyophilized, and characterized using enzymatic assays for hyaluronidase, phospholipase, and protease. Proteomic analysis of the venom was conducted employing bottom-up/shotgun approaches, SDS-PAGE, high-pH reversed-phase chromatography, and fractionation of tryptic peptides using nano-LC-MS/MS. The proteins found in H. h. horridum venom were reviewed according to the classification of the transcriptome previously reported. The proteomic approach identified 101 enzymes, 36 other proteins, 15 protein inhibitors, 11 host defense proteins, and 1 toxin, including novel venom components such as calcium-binding proteins, phospholipase A2 inhibitors, serpins, cathepsin, subtilases, carboxypeptidase-like, aminopeptidases, glycoside hydrolases, thioredoxin transferases, acid ceramidase-like, enolase, multicopper oxidases, phosphoglucose isomerase (PGI), fructose-1,6-bisphosphatase class 1, pentraxin-related, peptidylglycine α-hydroxylating monooxygenase/peptidyl-hydroxyglycine α-amidating lyase, carbonic anhydrase, acetylcholinesterase, dipeptidylpeptidase, and lysozymes. These findings contribute to understanding the venomous nature of H. h. horridum and highlight its potential as a source of bioactive compounds. Data are available via PRoteomeXchange with the identifier PXD052417.


Asunto(s)
Animales Ponzoñosos , Lagartos , Proteómica , Espectrometría de Masas en Tándem , Ponzoñas , Animales , Animales Ponzoñosos/genética , Animales Ponzoñosos/metabolismo , Hialuronoglucosaminidasa/metabolismo , Hialuronoglucosaminidasa/antagonistas & inhibidores , Hialuronoglucosaminidasa/genética , Hypocreales/química , Hypocreales/genética , Lagartos/genética , Lagartos/metabolismo , Proteoma/análisis , Proteómica/métodos , Proteínas de Reptiles/genética , Proteínas de Reptiles/metabolismo , Proteínas de Reptiles/química , Transcriptoma , Ponzoñas/química
2.
Cell Commun Signal ; 16(1): 45, 2018 08 14.
Artículo en Inglés | MEDLINE | ID: mdl-30107837

RESUMEN

BACKGROUND: In T cells, the Kv1.3 and the KCa3.1 potassium channels regulate the membrane potential and calcium homeostasis. Notably, during TEM cell activation, the number of Kv1.3 channels on the cell membrane dramatically increases. Kv1.3 blockade results in inhibition of Ca2+ signaling in TEM cells, thus eliciting an immunomodulatory effect. Among the naturally occurring peptides, the Vm24 toxin from the Mexican scorpion Vaejovis mexicanus is the most potent and selective Kv1.3 channel blocker known, which makes it a promissory candidate for its use in the clinic. We have shown that addition of Vm24 to TCR-activated human T cells inhibits CD25 expression, cell proliferation and reduces delayed-type hypersensitivity reactions in a chronic inflammation model. Here, we used the Vm24 toxin as a tool to investigate the molecular events that follow Kv1.3 blockade specifically on human CD4+ TEM cells as they are actively involved in inflammation and are key mediators of autoimmune diseases. METHODS: We combined cell viability, activation, and multiplex cytokine assays with a proteomic analysis to identify the biological processes affected by Kv1.3 blockade on healthy donors CD4+ TEM cells, following TCR activation in the presence or absence of the Vm24 toxin. RESULTS: The peptide completely blocked Kv1.3 channels currents without impairing TEM cell viability, and in response to TCR stimulation, it inhibited the expression of the activation markers CD25 and CD40L (but not that of CD69), as well as the secretion of the pro-inflammatory cytokines IFN-γ and TNF and the anti-inflammatory cytokines IL-4, IL-5, IL-9, IL-10, and IL-13. These results, in combination with data from the proteomic analysis, indicate that the biological processes most affected by the blockade of Kv1.3 channels in a T cell activation context were cytokine-cytokine receptor interaction, mRNA processing via spliceosome, response to unfolded proteins and intracellular vesicle transport, targeting the cell protein synthesis machinery. CONCLUSIONS: The Vm24 toxin, a highly specific inhibitor of Kv1.3 channels allowed us to define downstream functions of the Kv1.3 channels in human CD4+ TEM lymphocytes. Blocking Kv1.3 channels profoundly affects the mRNA synthesis machinery, the unfolded protein response and the intracellular vesicle transport, impairing the synthesis and secretion of cytokines in response to TCR engagement, underscoring the role of Kv1.3 channels in regulating TEM lymphocyte function.


Asunto(s)
Linfocitos T CD4-Positivos/efectos de los fármacos , Linfocitos T CD4-Positivos/metabolismo , Canal de Potasio Kv1.3/antagonistas & inhibidores , Péptidos/farmacología , Bloqueadores de los Canales de Potasio/farmacología , Receptores de Antígenos de Linfocitos T/metabolismo , Venenos de Escorpión/farmacología , Animales , Citocinas/biosíntesis
3.
J Nat Prod ; 81(8): 1899-1904, 2018 08 24.
Artículo en Inglés | MEDLINE | ID: mdl-30028606

RESUMEN

While most scorpion venom components identified in the past are peptidic or proteinic in nature, we report here a new alkaloid isolated from the venom of the Mexican scorpion Megacormus gertschi. Nuclear magnetic resonance and mass spectrometric investigations elucidate the structure of the alkaloid as ( Z)- N-(2-(1 H-imidazol-4-yl)ethyl)-3-(4-hydroxy-3-methoxyphenyl)-2-methoxyacrylamide (1). A chemical method of synthesizing this alkaloid is also described. Although abundant in venom, the above alkaloid was not found to have insecticidal activity. Structural analysis suggests that this venom alkaloid might be of potential interest for evaluating its medicinal effect.


Asunto(s)
Alcaloides/química , Venenos de Escorpión/química , Alcaloides/toxicidad , Animales , Cromatografía Líquida de Alta Presión , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Estructura Molecular , Escorpiones , Espectrometría de Masa por Ionización de Electrospray
4.
Biochim Biophys Acta ; 1838(9): 2140-8, 2014 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-24200946

RESUMEN

UyCT peptides are antimicrobial peptides isolated from the venom of the Australian scorpion. The activity of the UyCT peptides against Gram positive and Gram negative bacteria and red blood cells was determined. The membrane interactions of these peptides were evaluated by dye release (DR) of the fluorophore calcein from liposomes and isothermal titration calorimetry (ITC); and their secondary structure was determined by circular dichroism (CD). Three different lipid systems were used to mimic red blood cells, Escherichia coli and Staphylococcus aureus membranes. UyCT peptides exhibited broad spectrum antimicrobial activity with low MIC for S. aureus and multi-drug resistant Gram negative strains. Peptide combinations showed some synergy enhancing their potency but not hemolytic activity. The UyCT peptides adopted a helical structure in lipid environments and DR results confirmed that the mechanism of action is by disrupting the membrane. ITC data indicated that UyCT peptides preferred prokaryotic rather than eukaryotic membranes. The overall results suggest that UyCT peptides could be pharmaceutical leads for the treatment of Gram negative multiresistant bacterial infections, especially against Acinetobacter baumanni, and candidates for peptidomimetics to enhance their potency and minimize hemolysis. This article is part of a Special Issue entitled: Interfacially Active Peptides and Proteins. Guest Editors: William C. Wimley and Kalina Hristova.


Asunto(s)
Antiinfecciosos/química , Péptidos Catiónicos Antimicrobianos/química , Membrana Celular/efectos de los fármacos , Péptidos/química , Acinetobacter baumannii/efectos de los fármacos , Animales , Antiinfecciosos/farmacología , Péptidos Catiónicos Antimicrobianos/farmacología , Dicroismo Circular , Escherichia coli/efectos de los fármacos , Humanos , Membrana Dobles de Lípidos/química , Liposomas/química , Liposomas/metabolismo , Péptidos/farmacología , Estructura Secundaria de Proteína , Escorpiones/química , Staphylococcus aureus/efectos de los fármacos
6.
Biochim Biophys Acta ; 1830(6): 3427-36, 2013 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23403131

RESUMEN

BACKGROUND: Microbial antibiotic resistance is a challenging medical problem nowadays. Two scorpion peptides displaying antibiotic activity: hadrurin and vejovine were taken as models for the design of novel shorter peptides with similar activity. METHODS: Using the standard Fmoc-based solid phase synthesis technique of Merrifield twelve peptides (18 to 29 amino acids long) were synthesized, purified and assayed against a variety of multi-drug resistant Gram-negative bacteria from clinical isolates. Hemolytic and antiparasitic activities of the peptides and their possible interactions with eukaryotic cells were verified. Release of the fluorophore calcein from liposomes treated with these peptides was measured. RESULTS: A peptide with sequence GILKTIKSIASKVANTVQKLKRKAKNAVA), and three analogs: Δ(Α29), Δ(K12-Q18; Ν26-Α29), and K4N Δ(K12-Q18; Ν26-Α29) were shown to inhibit the growth of Gram-negative (E. coli ATCC25922) and Gram-positive bacteria (S. aureus), as well as multi-drug resistant (MDR) clinical isolated. The antibacterial and antiparasitic activities were found with peptides at 0.78 to 25µM and 5 to 25µM concentration, respectively. These peptides have low cytotoxic and hemolytic activities at concentrations significantly exceeding their minimum inhibitory concentrations (MICs), showing values between 40 and 900µM for their EC50, compared to the parent peptides vejovine and hadrurin that at the same concentration of their MICs lysed more than 50% of human erythrocytes cells. CONCLUSIONS: These peptides promise to be good candidates to combat infections caused by Gram-negative bacteria from nosocomial infections. GENERAL SIGNIFICANCE: Our results confirm that well designed synthetic peptides can be an alternative for solving the lack of effective antibiotics to control bacterial infections.


Asunto(s)
Antiinfecciosos , Antimaláricos , Farmacorresistencia Bacteriana Múltiple/efectos de los fármacos , Escherichia coli/crecimiento & desarrollo , Péptidos , Plasmodium berghei/crecimiento & desarrollo , Venenos de Escorpión , Staphylococcus aureus/crecimiento & desarrollo , Animales , Antiinfecciosos/síntesis química , Antiinfecciosos/química , Antiinfecciosos/farmacología , Antimaláricos/síntesis química , Antimaláricos/química , Antimaláricos/farmacología , Células COS , Chlorocebus aethiops , Células HEK293 , Humanos , Ratones , Ratones Endogámicos BALB C , Péptidos/síntesis química , Péptidos/química , Péptidos/farmacología
7.
Toxicon ; 249: 108062, 2024 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-39127082

RESUMEN

Envenomation by reptile venom, particularly from lizards, poses significant health risks and can lead to physiological and cardiovascular changes. The venom of Heloderma horridum horridum, endemic to Colima, Mexico, was tested on Wistar rats. Electrocardiographic (ECG) data were collected pre-treatment and at 5-min intervals for 1 h post-envenomation. A specially designed computational linear regression algorithm (LRA) was used for the segmentation analysis of the ECG data to improve the detection of fiducial points (P, Q, R, S, and T) in ECG waves. Additionally, heart tissue was analyzed for macroscopic and microscopic changes. The results revealed significant electrocardiographic alterations, including pacemaker migration, junctional extrasystoles, and intraventricular conduction aberrations. By applying a linear regression algorithm, the study compensated for noise and anomalies in the isoelectric line in an ECG signal, improving the detection of P and T waves and the QRS complex with an efficiency of 97.5%. Cardiac enzyme evaluation indicated no statistically significant differences between the control and experimental groups. Macroscopic and microscopic examination revealed no apparent signs of damage or inflammatory responses in heart tissues. This study enhances our understanding of the cardiovascular impact of Heloderma venom, suggesting a greater influence on changes in conduction and arrhythmias than on direct cardiac damage to the myocardium.


Asunto(s)
Algoritmos , Electrocardiografía , Ratas Wistar , Animales , Ratas , Modelos Lineales , Corazón/efectos de los fármacos , Lagartos , Masculino , Ponzoñas/toxicidad , México , Animales Ponzoñosos
9.
Toxins (Basel) ; 15(8)2023 08 09.
Artículo en Inglés | MEDLINE | ID: mdl-37624255

RESUMEN

Centruroides possanii is a recently discovered species of "striped scorpion" found in Mexico. Certain species of Centruroides are known to be toxic to mammals, leading to numerous cases of human intoxications in the country. Venom components are thought to possess therapeutic potential and/or biotechnological applications. Hence, obtaining and analyzing the secretory gland transcriptome and venom proteome of C. possanii is relevant, and that is what is described in this communication. Since this is a newly described species, first, its LD50 to mice was determined and estimated to be 659 ng/g mouse weight. Using RNA extracted from this species and preparing their corresponding cDNA fragments, a transcriptome analysis was obtained on a Genome Analyzer (Illumina) using the 76-base pair-end sequencing protocol. Via high-throughput sequencing, 19,158,736 reads were obtained and ensembled in 835,204 sequences. Of them, 28,399 transcripts were annotated with Pfam. A total of 244 complete transcripts were identified in the transcriptome of C. possanii. Of these, 109 sequences showed identity to toxins that act on ion channels, 47 enzymes, 17 protease inhibitors (PINs), 11 defense peptides (HDPs), and 60 in other components. In addition, a sample of the soluble venom obtained from this scorpion was analyzed using an Orbitrap Velos apparatus, which allowed for identification by liquid chromatography followed by mass spectrometry (LC-MS/MS) of 70 peptides and proteins: 23 toxins, 27 enzymes, 6 PINs, 3 HDPs, and 11 other components. Until now, this work has the highest number of scorpion venom components identified through omics technologies. The main novel findings described here were analyzed in comparison with the known data from the literature, and this process permitted some new insights in this field.


Asunto(s)
Escorpiones , Ponzoñas , Humanos , Animales , Ratones , Escorpiones/genética , Cromatografía Liquida , Espectrometría de Masas en Tándem , Inhibidores de Proteasas , Mamíferos
10.
Toxins (Basel) ; 15(2)2023 02 10.
Artículo en Inglés | MEDLINE | ID: mdl-36828459

RESUMEN

Skin secretions of toads are a complex mixture of molecules. The substances secreted comprise more than 80 different compounds that show diverse pharmacological activities. The compounds secreted through skin pores and parotid glands are of particular interest because they help toads to endure in habitats full of pathogenic microbes, i.e., bacteria, fungi, viruses, and protozoa, due to their content of components such as bufadienolides, alkaloids, and antimicrobial peptides. We carried out an extensive literature review of relevant articles published until November 2022 in ACS Publications, Google Scholar, PubMed, and ScienceDirect. It was centered on research addressing the biological characterization of the compounds identified in the species of genera Atelopus, Bufo, Duttaphrynus, Melanophryniscus, Peltopryne, Phrynoidis, Rhaebo, and Rhinella, with antibacterial, antifungal, antiviral, and antiparasitic activities; as well as studies performed with analogous compounds and skin secretions of toads that also showed these activities. This review shows that the compounds in the secretions of toads could be candidates for new drugs to treat infectious diseases or be used to develop new molecules with better properties from existing ones. Some compounds in this review showed activity against microorganisms of medical interest such as Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Coronavirus varieties, HIV, Trypanosoma cruzi, Leishmania chagasi, Plasmodium falciparum, and against different kinds of fungi that affect plants of economic interest.


Asunto(s)
Antiinfecciosos , Bufanólidos , Animales , Bufonidae , Antibacterianos , Bufanólidos/farmacología , Antifúngicos , Piel
11.
Toxicon ; 233: 107232, 2023 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-37536653

RESUMEN

Scorpions are a group of arthropods that strike fear in many people due to their severe medical symptoms, even death, caused by their venomous stings. Even so, not all scorpion species contain harmful venoms against humans but still have valuable bioactive molecules, which could be used in developing new pharmaceutical leads for treating important diseases. This work conducted a comprehensive analysis of the venom from the scorpion Thorellius intrepidus. The venom of T. intrepidus was separated by size exclusion chromatography, and four main fractions were obtained. Fraction IV (FIV) contained small molecules representing over 90% of the total absorbance at 280 nm. Analysis of fraction FIV by RP-HPLC indicated the presence of three main molecules (FIV.1, FIV.2, and FIV.3) with similar UV absorbance spectra profiles. The molecular masses of FIV.1, FIV.2, and FIV.3 were determined, resulting in 175.99, 190.07, and 218.16 Da, respectively. Further confirmation through 1H-NMR and 13C-NMR analyses revealed that these molecules were serotonin, N-methylserotonin, and bufotenidine. These intriguing compounds are speculated to play a pivotal role in self-defense and increasing venom toxicity and could also offer promising biotechnological applications as small bioactive molecules.


Asunto(s)
Picaduras de Escorpión , Venenos de Escorpión , Animales , Humanos , Escorpiones , Ponzoñas , Venenos de Escorpión/química
12.
Biochem Biophys Res Commun ; 429(1-2): 111-6, 2012 Dec 07.
Artículo en Inglés | MEDLINE | ID: mdl-23103547

RESUMEN

Scorpion γ-KTx toxins are important molecular tools for studying physiological and pharmacological functions of human ether-á-go-go related gene (hERG) K(+) channels. To pinpoint functional residues of this class of toxins involved in channel binding, we employed a combined approach that integrates evolutionary information and site-directed mutagenesis. Among three positively selected sites (PSSs) identified here, two (Gln18 and Met35) were found to be associated with the toxin's function because their changes significantly decreased the potency of ErgTx1 (also called CnErg1) on hERG1 channel. On the contrary, no potency alteration was observed at the third PSS (Ala42) when the mutation was introduced, which could be due to its location far from the functional surface of the toxin. Our strategy will accelerate the research of structure-function relationship of scorpion K(+) channel toxins.


Asunto(s)
Canales de Potasio Éter-A-Go-Go/antagonistas & inhibidores , Venenos de Escorpión/genética , Venenos de Escorpión/farmacología , Selección Genética , Secuencia de Aminoácidos , Animales , Células CHO , Cricetinae , Análisis Mutacional de ADN , Canal de Potasio ERG1 , Glutamina/genética , Humanos , Metionina/genética , Datos de Secuencia Molecular , Relación Estructura-Actividad
13.
Am J Cardiovasc Dis ; 11(3): 375-381, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34322307

RESUMEN

BACKGROUND: Computerized electrocardiogram (EKG) interpretation technology was developed in the mid-20th century, but its use continues to be controversial. This study aims to determine clinical factors which indicate greater odds of clinical significance of an abnormal computerized EKG interpretation. METHODS: The inclusion criteria for this retrospective study were patients who underwent outpatient echocardiography for the indication of an abnormal EKG and had an EKG abnormality diagnosed by the computerized EKG system. Qualifying patients had the results of their computerized EKG, echocardiogram, and charted patient characteristics collected. Computerized diagnoses and patient characteristics were assessed to determine if they were associated with increasing or decreasing the odds of an echocardiographic abnormality via logistic regression. Chi-square and t-test analyses were used for categorical and continuous variables, respectively. Odds ratios are presented as odds ratio [95% confidence interval]. A P-value of ≤ 0.05 was considered statistically significant. RESULTS: A total of 515 patients were included in this study. The population was 59% women with an average age of 57 ± 16 years, and a mean BMI of 30.1 ± 7.3 kg/m2. Patients with echocardiographic abnormalities tended to have more cardiac risk factors than patients without abnormalities. In our final odds ratio model consisting of both patient characteristics and EKG diagnoses, age, coronary disease (CAD), and diabetes mellitus (DM) increased the odds of an echocardiographic abnormality (1.04 [1.02-1.06], 2.68 [1.41-5.09], and 1.75 [1.01-3.04], respectively). That model noted low QRS voltage decreased the odds of an abnormal echocardiogram (0.31 [0.10-0.91]). CONCLUSION: Our findings suggest that in patients with an abnormal computerized EKG reading, the specific factors of older age, CAD, and DM are associated with higher odds of abnormalities on follow-up echocardiography. These results, plus practitioner overreading, can be used to determine more appropriate management when faced with an abnormal computerized EKG diagnosis.

14.
Peptides ; 141: 170553, 2021 07.
Artículo en Inglés | MEDLINE | ID: mdl-33862164

RESUMEN

Six peptides, belonging to the NDBP-4 family of scorpion antimicrobial peptides were structurally and functionally characterized. The sequence of the mature peptides VpCT1, VpCT2, VpCT3 and VpCT4 was inferred by transcriptomic analysis of the venom gland of the scorpion Mesomexovis variegatus. Analysis of their amino acid sequences revealed patterns that are also present in previously reported peptides that show differences in their hemolytic and antimicrobial activities in vitro. Two other variants, VpCT3W and VpCTConsensus were designed to evaluate the effect of sequence changes of interest on their structure and activity. The synthesized peptides were evaluated by circular dichroism to confirm their α-helical conformation in a folding promoting medium. The peptides were assayed on two Gram-positive and three Gram-negative bacterial strains, and on two yeast strains. They preferentially inhibited the growth of Staphylococcus aureus, were mostly ineffective on Pseudomonas aeruginosa, and moderately inhibited the growth of Candida yeasts. All six peptides exhibited hemolytic activity on human erythrocytes in the range of 4.8-83.7 µM. VpCT3W displayed increased hemolytic and anti-yeast activities, but showed no change in antibacterial activity, relative to its parental peptide, suggesting that Trp6 may potentiate the interaction of VpCT3 with eukaryotic cell membranes. VpCTConsensus showed broader and enhanced antimicrobial activity relative to several of the natural peptides. The results presented here contribute new information on the structure and function of NDBP-4 antimicrobial peptides and provides clues for the design of less hemolytic and more effective antimicrobial peptides.


Asunto(s)
Péptidos Antimicrobianos/química , Péptidos Antimicrobianos/farmacología , Escorpiones/química , Animales , Antibacterianos/química , Antibacterianos/farmacología , Antifúngicos/química , Antifúngicos/farmacología , Péptidos Antimicrobianos/síntesis química , Dicroismo Circular , Hemolíticos/química , Hemolíticos/farmacología , Humanos , Pruebas de Sensibilidad Microbiana , Relación Estructura-Actividad
15.
Toxicon ; 197: 114-125, 2021 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-33901550

RESUMEN

The peptide, denominated Ct1a, is a ß-toxin of 66 amino acids, isolated from venom of the scorpion, Centruroides tecomanus, collected in Colima, Mexico. This toxin was purified using size exclusion, cationic exchange, and reverse phase chromatography. It is the most abundant toxin, representing 1.7% of the soluble venom. Its molecular mass of 7588.9 Da was determined by mass spectrometry. The amino acid sequence was determined by Edman degradation and confirmed by transcriptomic analysis. Since neurons of the suprachiasmatic nucleus (SCN) maintain a spontaneous firing rate (SFR), we evaluated the physiological effects of toxin Ct1a on these neurons. The SFR exhibited a bimodal concentration-dependent response: 100 nM of Ct1a increased the SFR by 223%, whereas 500 nM and 1000 nM reduced it to 42% and 7%, respectively. Control experiments, consisting of recordings of the SFR during a time similar to that used in Ct1a testing, showed stability throughout the trials. Experiments carried out with denatured Ct1a toxin (500 nM) caused no variation in SFR recordings. Action potentials of SCN neurons, before and after Ct1a (100 nM) showed changes in the time constants of depolarization and repolarization phases, amplitude, and half-time. Finally, recordings of hNav1.6 sodium currents indicated that Ct1a shifts the channel activation to a more negative potential and reduces the amplitude of the peak current. These results all demonstrate that toxin Ct1a affects the SFR of SCN neurons by acting upon sodium channels of sub-type 1.6, implicating them in regulation of the SFR of SCN neurons.


Asunto(s)
Venenos de Escorpión , Escorpiones , Animales , México , Neuronas , Núcleo Supraquiasmático , Ponzoñas
16.
Toxicon ; 179: 21-32, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32126222

RESUMEN

Centruroides hirsutipalpus (Scorpiones: Buthidae) is related to the "striped scorpion" group inhabiting the western Pacific region of Mexico. Human accidents caused by this species are medically important due to the great number of people stung and the severity of the resulting intoxication. This communication reports an extensive venom characterization using high-throughput proteomic and Illumina transcriptomic sequencing performed with RNA purified from its venom glands. 2,553,529 reads were assembled into 44,579 transcripts. From these transcripts, 23,880 were successfully annoted using Trinotate. Using specialized databases and by performing bioinformatic searches, it was possible to identify 147 putative venom protein transcripts. These include α- and ß-type sodium channel toxins (NaScTx), potassium channel toxins (KScTx) (α-, ß-, δ-, γ- and λ-types), enzymes (metalloproteases, hyaluronidases, phospholipases, serine proteases, and monooxygenases), protease inhibitors, host defense peptides (HDPs) such as defensins, non-disulfide bridge peptides (NDBPs), anionic peptides, superfamily CAP proteins, insulin growth factor-binding proteins (IGFBPs), orphan peptides, and other venom components (La1 peptides). De novo tandem mass spectrometric sequencing of digested venom identificatied 50 peptides. The venom of C. hirsutipalpus contains the highest reported number (77) of transcripts encoding NaScTxs, which are the components responsible for human fatalities.


Asunto(s)
Venenos de Escorpión/química , Escorpiones , Animales , Proteínas de Artrópodos/metabolismo , Glándulas Exocrinas , Secuenciación de Nucleótidos de Alto Rendimiento , México , Proteoma/metabolismo , Proteómica , Venenos de Escorpión/metabolismo , Transcriptoma/fisiología
17.
Toxicon ; 180: 62-78, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32283106

RESUMEN

Lizards of the Helodermatidae (Anguimorpha) family consist of at least two well recognized species: Heloderma horridum horridum and Heloderma suspectum suspectum. They contain specialized glands in their jaws that produce venomous secretions that causes envenoming symptoms to bitten animals. One way to study proteins from such secretions is by RNA-seq; a powerful molecular tool to characterize the transcriptome of such specialized gland, and its protein secretions. The total RNA from venom gland tissues of H. horridum horridum was extracted and a cDNA library was constructed and sequenced. Overall, 114,172 transcripts were found, and 199 were annotated based on sequence similarities to previously described peptides/proteins. Transcripts coding for putative exendins, defensins, natriuretics and serine protease inhibitors were the most highly expressed. Transcripts that code for several putative serine proteases, phospholipases, metalloproteases, lipases, L-amino oxidase and nucleases were also found. Some of the novel identified transcripts were translationally controlled tumor proteins, venom factors, vespryns, waprins, lectins, cystatins and serine protease inhibitors. All these new protein structures may contribute to a better understanding of the venomous secretions of the Helodermatidae family.


Asunto(s)
Lagartos/genética , Ponzoñas , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Secuenciación de Nucleótidos de Alto Rendimiento , Lagartos/metabolismo , Péptidos , Fosfolipasas , Transcriptoma
18.
Vive (El Alto) ; 6(16): 322-336, abr. 2023.
Artículo en Español | LILACS | ID: biblio-1442258

RESUMEN

Las normativas legales peruanas como es la Ley 29973, busca la promoción, protección de las personas con discapacidad en el ámbito laboral logrando así el desarrollo, igualdad e inclusión social, política que actualmente aplicado en las instituciones públicas ni privadas en Perú. Objetivo. Analizar la influencia de la discapacidad física en la inserción laboral en Perú. Materiales y Métodos. La metodología que se utilizó estuvo enmarcada en el enfoque cuantitativo, el tipo de investigación fue básica causal explicativo, el método hipotético deductivo, el diseño de la investigación fue no experimental de corte transeccional. La población fue de 61 usuarios, la técnica utilizada para la recolección de datos fue la encuesta y como instrumento el cuestionario, los datos se analizaron mediante el software estadístico SPSS. El procesamiento estadístico fue no paramétrico de Chi-Cuadrada. Resultados. Como resultado de la investigación fue que el 31,1% de los usuarios manifestaron que no están insertados en el mercado laboral ya que la discapacidad física que poseen es de tipo paraplejia, es decir, que, ambas piernas carecen de movilidad lo cual es una restricción para su desarrollo y dificulta su integración en el mercado laboral. Conclusiones. La discapacidad influye significativamente en la inserción laboral de los usuarios de la Oficina Municipal de Atención a las Personas con Discapacidad.


Peruvian legal regulations such as Law 29973, seeks the promotion and protection of people with disabilities in the workplace, thus achieving development, equality and social inclusion, a policy that is currently applied in public and private institutions in Peru. Objective. To analyze the influence of physical disability on labor market insertion in Peru. Materials and Methods. The methodology used was framed in the quantitative approach, the type of research was basic causal explanatory, the method was hypothetical deductive, the research design was non-experimental of transectional cut. The population consisted of 61 users, the technique used for data collection was the survey and the instrument was the questionnaire; the data were analyzed using SPSS statistical software. The statistical processing was nonparametric Chi-Square. Results. As a result of the research, 31.1% of the users stated that they are not inserted in the labor market since the physical disability they have is paraplegia, that is, both legs lack mobility, which is a restriction for their development and hinders their integration in the labor market. Conclusions. Disability has a significant influence on the labor market insertion of the users of the Municipal Office of Attention to People with Disabilities.


As normas legais peruanas, como a Lei 29973, buscam a promoção e a proteção de pessoas com deficiência no local de trabalho, alcançando assim o desenvolvimento, a igualdade e a inclusão social, uma política que atualmente é aplicada em instituições públicas e privadas no Peru. Objetivo. Analisar a influência da deficiência física na inserção laboral no Peru. Materiais e métodos. A metodologia utilizada foi enquadrada no enfoque quantitativo, o tipo de pesquisa foi causal explicativa básica, o método foi hipotético dedutivo, o desenho da pesquisa foi de corte transeccional não experimental. A população foi de 61 usuários, a técnica utilizada para a coleta de dados foi a pesquisa e como instrumento o questionário, os dados foram analisados com o uso do software estatístico SPSS. O processamento estatístico foi o qui-quadrado não paramétrico. Resultados. Como resultado da pesquisa, 31,1% dos usuários afirmaram que não estão inseridos no mercado de trabalho porque a deficiência física que possuem é a paraplegia, ou seja, falta mobilidade nas duas pernas, o que é uma restrição para o seu desenvolvimento e dificulta a sua integração no mercado de trabalho. Conclusões. A deficiência tem uma influência significativa na inserção no mercado de trabalho dos usuários da Secretaria Municipal de Atenção às Pessoas com Deficiência.


Asunto(s)
Encuestas y Cuestionarios , Personas con Discapacidad , Mercado de Trabajo
19.
Toxins (Basel) ; 10(9)2018 09 05.
Artículo en Inglés | MEDLINE | ID: mdl-30189638

RESUMEN

To understand the diversity of scorpion venom, RNA from venomous glands from a sawfinger scorpion, Serradigitus gertschi, of the family Vaejovidae, was extracted and used for transcriptomic analysis. A total of 84,835 transcripts were assembled after Illumina sequencing. From those, 119 transcripts were annotated and found to putatively code for peptides or proteins that share sequence similarities with the previously reported venom components of other species. In accordance with sequence similarity, the transcripts were classified as potentially coding for 37 ion channel toxins; 17 host defense peptides; 28 enzymes, including phospholipases, hyaluronidases, metalloproteases, and serine proteases; nine protease inhibitor-like peptides; 10 peptides of the cysteine-rich secretory proteins, antigen 5, and pathogenesis-related 1 protein superfamily; seven La1-like peptides; and 11 sequences classified as "other venom components". A mass fingerprint performed by mass spectrometry identified 204 components with molecular masses varying from 444.26 Da to 12,432.80 Da, plus several higher molecular weight proteins whose precise masses were not determined. The LC-MS/MS analysis of a tryptic digestion of the soluble venom resulted in the de novo determination of 16,840 peptide sequences, 24 of which matched sequences predicted from the translated transcriptome. The database presented here increases our general knowledge of the biodiversity of venom components from neglected non-buthid scorpions.


Asunto(s)
Proteínas de Artrópodos/análisis , Venenos de Escorpión/química , Animales , Bloqueadores de los Canales de Calcio/análisis , Bloqueadores de los Canales de Calcio/química , Femenino , Perfilación de la Expresión Génica , Hialuronoglucosaminidasa/análisis , Hialuronoglucosaminidasa/química , Masculino , Péptido Hidrolasas/análisis , Péptido Hidrolasas/química , Péptidos/análisis , Péptidos/química , Fosfolipasas A2/análisis , Fosfolipasas A2/química , Bloqueadores de los Canales de Potasio/análisis , Bloqueadores de los Canales de Potasio/química , Proteoma , Proteómica , Escorpiones , Bloqueadores de los Canales de Sodio/análisis , Bloqueadores de los Canales de Sodio/química
20.
Toxicon ; 151: 47-62, 2018 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-29964058

RESUMEN

The recent publication of high-throughput transcriptomic and proteomic analyses of scorpion venom glands has increased our knowledge on the biodiversity of venom components. In this contribution, we report the transcriptome of the venom gland and the proteome of the venom for the scorpion species Paravaejovis schwenkmeyeri, a member of the family Vaejovidae. We report 138 annotated transcripts encoding putative peptides/proteins with sequence identity to known venom components available from different databases. A fingerprint analysis containing the molecular masses of 212 components of the whole soluble venom revealed molecular weights of approximately 700 to 13,800 Da, with most detected proteins ranging from 1500 to 3000 Da. Amino acid sequencing of venom components by LC-MS/MS allowed the identification of fragments from 27 peptides encoded by transcripts found in the transcriptome analysis. Enzymatic assays conducted with the soluble venom fraction confirmed the presence of enzymes such as hyaluronidases and phospholipases. The database presented here increases our general knowledge on the biodiversity of venom components from neglected non-buthid scorpions.


Asunto(s)
Proteínas de Artrópodos/metabolismo , Proteoma , Venenos de Escorpión/química , Escorpiones/fisiología , Transcriptoma , Secuencia de Aminoácidos , Animales , Proteínas de Artrópodos/química , Proteínas de Artrópodos/genética , Biología Computacional , Canales Iónicos/antagonistas & inhibidores , Toxinas Biológicas/química , Toxinas Biológicas/genética , Toxinas Biológicas/metabolismo
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