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1.
Animals (Basel) ; 13(11)2023 Jun 02.
Artículo en Inglés | MEDLINE | ID: mdl-37889796

RESUMEN

This study determined the effect of pelagic Sargassum on in vitro dry matter and organic matter degradation, total gas production (TGP), and protozoa population. The treatments were different levels of Sargassum inclusion on a basal substrate (Stargrass hay; Cynodon nlemfuensis) as follows: T0 (control treatment based on Stargrass hay), T10 (90% Stargrass hay + 10% Sargassum), T20 (80% Stargrass hay + 20% Sargassum), and T30 (70% Stargrass hay + 30% Sargassum). Ruminal fermentation kinetics and protozoa population were determined during 72 h of in vitro incubations. Compared to control, dry matter degradability at 48 and 72 h and organic matter degradability at 24 and 48 h were higher in Sargassum treatments. TGP was lower with T20 at 48 h. The total population of protozoa and the concentration of Entodinium spp. were lower at T20 at 48 h and T30 at 72 h. Cl, S, Ca, K, and Zn (103, 5.97, 88.73, 285.70 g/kg, and 15,900 mg/kg) were high in Sargassum, reaching twice or even nine times higher than the contents in Stargrass (11.37, 1.60, 43.53, 87.73 g/kg, and 866.67 mg/kg). Overall, up to 30% pelagic Sargassum could be included in hay-based substrates from tropical grasses without negative effects on in vitro dry matter and organic matter degradability.

2.
Animals (Basel) ; 12(5)2022 Feb 24.
Artículo en Inglés | MEDLINE | ID: mdl-35268141

RESUMEN

This study was carried out to evaluate the residual feed intake (RFI), volatile fatty acid (VFA) production and enteric methane (CH4) from growing Pelibuey sheep. In this case, 12 non-castrated Pelibuey with an initial average live weight (LW) of 21.17 ± 3.87 kg and an age of 3 months, were housed in individual pens and fed a basal diet with 16% of crude protein and 11 MJ ME for 45 days. Dry matter intake (DMI) was measured and the daily weight gain (DWG) was calculated using a linear regression between the LW and experimental period. Mean metabolic live weight (LW0.75) was calculated. RFI was determined by linear regression with DWG and LW0.75 as independent variables. Lambs were classified as low, medium, and high RFI. Feed efficiency was determined as DWG/DMI. For determining rumen pH, ammonia nitrogen concentration NH3-N), and VFA, ruminal fluid was obtained using an esophageal probe on day 40. Feed intake of low RFI lambs was approximately 16% lower (p < 0.05) while growth rate was not significantly different. Their average energy loss, expressed as CH4 production per kilogram of metabolic weight, was 17% lower (p < 0.05).

3.
Animals (Basel) ; 10(7)2020 Jun 29.
Artículo en Inglés | MEDLINE | ID: mdl-32610660

RESUMEN

The objective this study was to evaluate the effect of different doses of Moringa oleifera leaf extract (MOE) on milk production and milk composition in ewes and on preweaning performance of their lambs. Twenty-four lactating ewes were housed individually with their lambs and assigned to four groups in a completely randomized design. The treatments included a basal diet without MOE (MOE0) or a basal diet supplemented with either 20 mL MOE per ewe per day (MOE20), 40 mL MOE per ewe per day (MOE40) or 60 mL MOE per ewe per day (MOE60). Over 45 days, milk production was recorded weekly and individual milk samples were collected for chemical analysis. Milk yield, fat-corrected milk and daily yields were similar among the four treatments. The supply of MOE did not affect ewe weaning efficiency and average daily gain or litter weaning weight of the lambs. Overall, the results from this study showed that dietary supplementation of hydroalcoholic extracts of Moringa oleifera leaves at doses of 20, 40 or 60 mL/ewes/d in lactating ewes does not have negative effects on milk yield, milk composition or lamb performance.

4.
Animals (Basel) ; 10(3)2020 Mar 20.
Artículo en Inglés | MEDLINE | ID: mdl-32244883

RESUMEN

The study aimed to evaluate the relationship between udder measurements and milk yield (MY) in dairy Pelibuey ewes. Udder measurements were taken twice a week for eight weeks before (initial) and after (final) milking, including udder depth (UD), udder circumference (UC), udder width (UW), teat length (TL) and teat diameter (TD) in 38 multiparous ewes. Additionally, udder volume (UV) and the difference (VDF) between initial UV (UVi) and final (UVf) was calculated as VDF = UVi - UVf. The MY varied from 0.10 kg/d to 1.04 kg/d, with a mean of 0.39 kg/d, ± 0.18 kg/d. Initial UC (UCi) ranged from 25.80 cm to 53.30 cm, and VDF varied from 1 cm3 to 2418 cm3. The TL and TD were not correlated with MY (p > 0.05), while UCi, UVi and VDF were positively correlated with MY (p < 0.0001; r = from 0.66 to 0.74). For the prediction of MY, the obtained equations had an r2 ranging from 0.54 to 0.63. The UCi, UDf, UWi and UWf were included in these models (p < 0.05). It is concluded that there was an acceptable correlation (r = 0.60) between the measurements of the udder, the volume of the udder and the daily milk yield in Pelibuey sheep. When direct measurements of milk production cannot be performed in practice, the measurement of udders and their volume could be a viable alternative to estimate milk yield production as an indirect method.

5.
Trop Anim Health Prod ; 52(5): 2341-2347, 2020 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-32162187

RESUMEN

The aim of the present study was to develop and evaluate an equation to predict body weight (BW) using hip width (HW) in Pelibuey ewe lambs and ewes. Five hundred seventy-seven 2-month-old to 3-year-old, non-pregnant, non-lactating, clinically healthy ewe lambs and adult ewes with a mean BW of 34.7 ± 12.4 kg and HW of 15.6 ± 3.4 cm were considered. Three equations were evaluated: BW (kg): - 19.17 + 3.46 × HW (Eq. 1), BW (kg): - 17.79 + 3.25 × HW + 0.007 × HW2 (Eq. 2) and BW (kg): 0.39 × HW1.63 (Eq. 3). Independent data from 80 animals with similar characteristics (BW of 23.4 ± 10.9 kg and HW of 12 ± 3.1 cm) were also considered to evaluate the developed equations. The evaluation was based on the relationship between the observed and predicted values of BW analysed using a linear regression, the mean squared error of prediction (MSEP), the root MSEP (RMSEP) and the concordance correlation coefficients (CCCs). Additionally, cross-validation analyses were performed using the k-folds validation (k = 10) procedure. The correlation coefficient (r) between BW and HW was 0.94 (P < 0.001). The parameters for precision and accuracy showed that the proposed equations had high precision (R2 > 0.95%), accuracy (Cb > 0.98) and reproducibility (CCC > 0.96) in predicting the BW of ewe lambs and adult ewes. Equation (1) accurately predicted observed BW, with a bias (observed - predicted) of 4.3 kg and RMSEP of 9.68% with respect to the observed BW (random error of 84.23%); it also generated the best prediction according to the residual mean squared prediction error, coefficient of determination and mean absolute error. In conclusion, the highly correlated relationship between BW and HW in Pelibuey ewe lambs and adult ewes under humid tropic conditions enabled the development of mathematical models herein to estimate BW with an adequate goodness of fit. The linear model showed the best performance according to the goodness-of-fit evaluation and internal and external validation; hence, this model is proposed for use in both the experimental and commercial farms.


Asunto(s)
Peso Corporal , Ovinos/fisiología , Algoritmos , Animales , Femenino , Modelos Lineales , Modelos Biológicos , Reproducibilidad de los Resultados
6.
Toxins (Basel) ; 11(2)2019 02 20.
Artículo en Inglés | MEDLINE | ID: mdl-30791616

RESUMEN

Californiconus californicus, previously named Conus californicus, has always been considered a unique species within cone snails, because of its molecular, toxicological and morphological singularities; including the wide range of its diet, since it is capable of preying indifferently on fish, snails, octopus, shrimps, and worms. We report here a new cysteine pattern conotoxin assigned to the O1-superfamily capable of inhibiting the growth of Mycobacterium tuberculosis (Mtb). The conotoxin was tested on a pathogen reference strain (H37Rv) and multidrug-resistant strains, having an inhibition effect on growth with a minimal inhibitory concentration (MIC) range of 3.52⁻0.22 µM, similar concentrations to drugs used in clinics. The peptide was purified from the venom using reverse phase high-performance liquid chromatography (RP-HPLC), a partial sequence was constructed by Edman degradation, completed by RACE and confirmed with venom gland transcriptome. The 32-mer peptide containing eight cysteine residues was named O1_cal29b, according to the current nomenclature for this type of molecule. Moreover, transcriptomic analysis of O-superfamily toxins present in the venom gland of the snail allowed us to assign several signal peptides to O2 and O3 superfamilies not described before in C. californicus, with new conotoxins frameworks.


Asunto(s)
Antibacterianos/farmacología , Conotoxinas/farmacología , Mycobacterium tuberculosis/efectos de los fármacos , Péptidos/farmacología , Animales , Conotoxinas/genética , Caracol Conus , Farmacorresistencia Bacteriana/efectos de los fármacos , Resistencia a Múltiples Medicamentos/efectos de los fármacos , Mycobacterium tuberculosis/crecimiento & desarrollo , Péptidos/genética , Tuberculosis Resistente a Múltiples Medicamentos
7.
Mar Drugs ; 16(4)2018 Mar 31.
Artículo en Inglés | MEDLINE | ID: mdl-29614715

RESUMEN

Variable new antigen receptor domain (vNAR) antibodies are novel, naturally occurring antibodies that can be isolated from naïve, immune or synthetic shark libraries. These molecules are very interesting to the biotechnology and pharmaceutical industries because of their unique characteristics related to size and tissue penetrability. There have been some approved anti-angiogenic therapies for ophthalmic conditions, not related to vNAR. This includes biologics and chimeric proteins that neutralize vascular endothelial growth factor (VEGF)165, which are injected intravitreal, causing discomfort and increasing the possibility of infection. In this paper, we present a vNAR antibody against human recombinant VEGF165 (rhVEGF165) that was isolated from an immunized Heterodontus francisci shark. A vNAR called V13, neutralizes VEGF165 cytokine starting at 75 µg/mL in an in vitro assay based on co-culture of normal human dermal fibroblasts (NHDFs) and green fluorescence protein (GFP)-labeled human umbilical vein endothelial cells (HUVECs) cells. In the oxygen-induced retinopathy model in C57BL/6:Hsd mice, we demonstrate an endothelial cell count decrease. Further, we demonstrate the intraocular penetration after topical administration of 0.1 µg/mL of vNAR V13 by its detection in aqueous humor in New Zealand rabbits with healthy eyes after 3 h of application. These findings demonstrate the potential of topical application of vNAR V13 as a possible new drug candidate for vascular eye diseases.


Asunto(s)
Productos Biológicos/farmacocinética , Enfermedades de la Retina/tratamiento farmacológico , Tiburones , Anticuerpos de Dominio Único/farmacología , Factor A de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Administración Tópica , Animales , Productos Biológicos/inmunología , Productos Biológicos/aislamiento & purificación , Productos Biológicos/uso terapéutico , Células Cultivadas , Técnicas de Cocultivo , Modelos Animales de Enfermedad , Ojo/irrigación sanguínea , Ojo/metabolismo , Fibroblastos , Células Endoteliales de la Vena Umbilical Humana , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Soluciones Oftálmicas/farmacocinética , Soluciones Oftálmicas/uso terapéutico , Oxígeno/toxicidad , Conejos , Proteínas Recombinantes/metabolismo , Enfermedades de la Retina/inducido químicamente , Enfermedades de la Retina/patología , Anticuerpos de Dominio Único/inmunología , Anticuerpos de Dominio Único/aislamiento & purificación , Anticuerpos de Dominio Único/uso terapéutico , Factor A de Crecimiento Endotelial Vascular/inmunología , Factor A de Crecimiento Endotelial Vascular/metabolismo
8.
Toxins (Basel) ; 9(3)2017 02 23.
Artículo en Inglés | MEDLINE | ID: mdl-28241514

RESUMEN

Scorpion stings on humans are medically relevant because they may contain toxins that specifically target ion channels. During antivenom production, pharmaceutical companies must use a large number of experimental animals to ensure the antivenom's efficacy according to pharmacopeia methods. Here we present an electrophysiological alternative for the evaluation of horse antivenoms produced against two species of Moroccan scorpions: Buthus mardochei and Androctonus mauretanicus. Human sodium and potassium channels and acetylcholine nicotinic receptors were analyzed by standard patch-clamp techniques. The results showed that the antivenom is capable of reversing ion current disruption caused by the venom application. We propose the use of this in vitro technique for antivenom evaluation as an alternative to using a large number of live animals.


Asunto(s)
Antivenenos/farmacología , Venenos de Escorpión/toxicidad , Canales de Sodio/fisiología , Alternativas a las Pruebas en Animales , Animales , Fenómenos Electrofisiológicos , Células HEK293 , Humanos , Escorpiones
9.
Medisan ; 20(9)set. 2016. tab
Artículo en Español | LILACS, CUMED | ID: lil-797484

RESUMEN

Se realizó un estudio descriptivo y transversal, a fin de evaluar el estado nutricional de 91 pacientes egresados del Servicio de Cirugía General del Hospital Provincial Docente Clinicoquirúrgico "Saturnino Lora Torres" de Santiago de Cuba, quienes habían sido intervenidos con cirugía mayor electiva, durante el trienio 2013-2015. Se halló un decrecimiento de los pacientes con malnutrición y aumento de los valores de bioimpedancia. Esta última constituyó el método más sensible para determinar los afectados con posibilidad de complicarse, pues 89,5 % de los malnutridos tuvieron complicaciones en el periodo posoperatorio. Asimismo, la infección del sitio quirúrgico fue la complicación más frecuente. Se concluyó que la malnutrición es un importante problema para los pacientes operados y los métodos más idóneos para su valoración nutricional son el índice de masa corporal, la bioimpedancia, así como el conteo global de linfocitos.


A descriptive and cross-sectional study was carried out, in order to evaluate the nutritional state of 91 patients discharged from the General Surgery Service of "Saturnino Lora Torres" Clinical Surgical Teaching Provincial Hospital in Santiago de Cuba who had been treated with elective major surgery, during the triennium 2013-2015. There was a decrease of the patients with malnutrition and increase of the bioimpedance values, which constituted the most sensitive method to determine those patients affected with possibility of complications, because 89.5% of the malnourished cases had complications in the postoperative period. Also, the infection of the surgical site was the most frequent complication. It was concluded that malnutrition is an important problem for the operated patients and the most suitable methods for their nutritional evaluation are the corporal mass index, bioimpedance, as well as the global count of lymphocytes.


Asunto(s)
Procedimientos Quirúrgicos Operativos , Evaluación Nutricional , Complicaciones Posoperatorias , Atención Secundaria de Salud
10.
Acta otorrinolaringol. cir. cabeza cuello ; 42(1): 18-22, ene.-mar. 2014. ilus
Artículo en Español | LILACS | ID: lil-746370

RESUMEN

Introducción: El examen físico no ha demostrado una predicción diagnóstica significativa en la apnea obstructiva del sueño (AOS). Aunque la polisomnografía es el estándar de oro, sus costos y disponibilidad la hacen difícil de realizar en algunos casos; así, es importante encontrar otras herramientas que brinden una aproximación referente a la severidad de la AOS. Objetivo: Establecer la relación entre la circunferencia del cuello y los parámetros polisomnográficos. Métodos: Estudio longitudinal, prolectivo, observacional, analítico en una cohorte prospectiva. Se reclutaron pacientes con sospecha de AOS en la consulta de Otorrinolaringología; se les practicó polisomnografía y medición de la circunferencia del cuello (CC). Se realizó prueba T para muestras independientes y parámetros descriptivos en el análisis estadístico. Resultados: Se evaluaron 141 pacientes, 95 hombres (67%); CC, 30-59 cm; saturación mínima de oxígeno, 35-91%; saturación promedio con rango de 65-96%; índice de apnea-hipopnea, 0,6-173 eventos/hora. Se encontraron diferencias significativas, p < 0,05, entre los parámetros medidos en los pacientes con circunferencia del cuello mayor de 43 cm. Conclusión: La CC mayor de 43 cm se asocia con peores resultados en los parámetros polisomnográficos, lo cual indica la asociación de la CC con la severidad de la AOS...


Introduction: Physical assessment hasn’t shown a very strong predictive severity diagnostic value in the obstructive sleep apnea (OSA). Polysomnography is the gold standard diagnostic tool, but it is expensive and difficult to get it in some cases. Therefore it is important to find other diagnostic studies that may give us an approximation to the severity of OSA. Objective: To establish the relationship between neck circumference and polysomnography parameters. Methods: This was a longitudinal - observational and analytic study in a population-based prospective and prolective cohort. We collected the patients with suspicion of OSA, in the Otolaryngology clinics; they were assessed with the neck circumference measure and diagnostic Polysomnography. We determined the Independent samples T- Test, and the descriptive parameters in the statistical analysis. Results: We evaluated 141 patients, 95 males (67,4%), neck circumference (NC) range: 30 – 59 cm; minimal oxygen saturation range: 35– 91% with a mean of 72%; mean oxygen saturation range: 65 – 96% with a mean of 88,9%; apnea hipopnea index (AHI) range: 0,6 – 173 events/hour with a mean 52,6 events/hour. There is a statistical significance p value lesser than 0,05 between the NC greater than 43 cm and NC lesser than 43 cm group, in these parameters: minimal saturation, mean saturation and AHI. Conclusion: The NC greater than 43 cm was associated with the worse results in the following polysomnography parameters: minimal saturation, mean saturation and AHI. These findings may give us an association between NC and the severity of the OSA...


Asunto(s)
Humanos , Apnea Obstructiva del Sueño , Apnea Obstructiva del Sueño/diagnóstico , Cuello/fisiología
11.
BMC Immunol ; 14: 17, 2013 Apr 02.
Artículo en Inglés | MEDLINE | ID: mdl-23548047

RESUMEN

BACKGROUND: In sepsis, tumor necrosis factor (TNF) is the key factor triggering respiratory burst, tissue injury and disseminated coagulation. Anti-TNF strategies based on monoclonal antibodies or F(ab')2 fragments have been used in sepsis with contradictory results. Immunoglobulin new antigen receptors (IgNAR) are a unique subset of antibodies consisting of five constant (CNAR) and one variable domains (VNAR). VNAR domains are the smallest, naturally occurring, antibody-based immune recognition units, having potential use as therapy. Our aim was to explore the impact of an anti-TNF VNAR on survival in an experimental model of endotoxic shock. Also, mRNA expression and serum protein of several inflammatory molecules were measured. RESULTS: Endotoxic shock was induced by lipopolysaccharide (LPS) in male Balb/c mice. Animals were treated with anti-TNF VNAR domains, F(ab')2 antibody fragments, or saline solution 15 minutes before, 2 h and 24 h after lethal dose100 (LD100) LPS administration. TNF blockade with either VNAR domains or F(ab')2 fragments were associated with lower mortality (60% and 75%, respectively) compared to LD100. Challenge with LPS induced significant production of serum TNF and interleukins -10 and -6 at 3 h. After that, significant reduction of IL-6 at 24 h (vs 3 h) was shown only in the VNAR group. Nitrites level also increased in response to LPS. In liver, TNF and IL-10 mRNA expression showed a pro-inflammatory imbalance in response to LPS. Blocking TNF was associated with a shift towards an anti-inflammatory status; however, polarization was more pronounced in animals receiving F(ab')2 fragments than in those with VNAR therapy. With regard to IL-6, gene expression was increased at 3 h in all groups. TNF blockade was associated with rapid and sustained suppression of IL-6 expression, even more evident in the VNAR group. Finally, expression of inducible-nitric oxide synthase (iNOS) increased in response to LPS at 3 h, but this was decreased at 24 h only in the anti-TNF VNAR group. CONCLUSIONS: Anti-TNF VNAR single domains improved survival in a murine model of endotoxic shock. Protection was associated with regulation in the TNF/IL-10 balance, attenuation of IL-6 and iNOS gene expression in the liver as well as decreased serum IL-6 concentration.


Asunto(s)
Inflamación/complicaciones , Inflamación/tratamiento farmacológico , Choque Séptico/complicaciones , Choque Séptico/tratamiento farmacológico , Anticuerpos de Dominio Único/uso terapéutico , Factor de Necrosis Tumoral alfa/inmunología , Animales , Biomarcadores/sangre , Modelos Animales de Enfermedad , Humanos , Inflamación/sangre , Inflamación/patología , Interleucina-10/metabolismo , Interleucina-6/metabolismo , Lipopolisacáridos , Hígado/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Nitratos/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Estructura Terciaria de Proteína , ARN Mensajero/genética , ARN Mensajero/metabolismo , Choque Séptico/sangre , Análisis de Supervivencia , Resultado del Tratamiento
12.
MAbs ; 5(1): 80-5, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-23221782

RESUMEN

The therapeutic use of single domain antibodies (sdAbs) is a promising new approach because these small antibodies maintain antigen recognition and neutralization capacity, have thermal and chemical stability and have good solubility. In this study, using phage display technology, we isolated a variable domain of a IgNAR (vNAR) from a Heterodontus francisci shark immunized against the recombinant human cytokine TNFα (rhTNFα). One clone T43, which expresses the vNAR protein in the periplasmic space, was isolated from the fourth round of panning. T43 had the capacity to recognize rhTNF and neutralize it in vitro, indicating that T43 has potential as a therapeutic that can be used for diseases in which this pro-inflammatory cytokine needs to be controlled.


Asunto(s)
Anticuerpos Neutralizantes/inmunología , Tiburones/inmunología , Anticuerpos de Dominio Único/inmunología , Factor de Necrosis Tumoral alfa/inmunología , Secuencia de Aminoácidos , Animales , Anticuerpos Neutralizantes/biosíntesis , Anticuerpos Neutralizantes/química , Anticuerpos Neutralizantes/uso terapéutico , Especificidad de Anticuerpos , Humanos , Inmunización , Región Variable de Inmunoglobulina/biosíntesis , Región Variable de Inmunoglobulina/química , Región Variable de Inmunoglobulina/inmunología , Inmunoglobulinas/biosíntesis , Inmunoglobulinas/química , Inmunoglobulinas/inmunología , Masculino , Ratones , Datos de Secuencia Molecular , Pruebas de Neutralización , Biblioteca de Péptidos , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Choque Séptico/inmunología , Anticuerpos de Dominio Único/biosíntesis , Anticuerpos de Dominio Único/química , Anticuerpos de Dominio Único/uso terapéutico , Factor de Necrosis Tumoral alfa/genética
13.
Toxicon ; 52(8): 881-8, 2008 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-18926842

RESUMEN

As a response to the antivenom shortage in Sub-Saharan Africa, evident for well over a decade, we developed a new polyvalent anti-ophidian antivenom (Antivipmyn((R)) Africa) designed for use in the region. We report a detailed characterization of its biochemical composition (protein content and profiling by size-exclusion chromatography and electrophoresis) as well as the specific and para-specific neutralization potencies (as median effective dose in the mouse lethality test). Additionally, we studied the neutralization of hemorrhagic, anti-hemostatic and necrotic activities of Echis ocellatus venom, responsible for a majority of severe envenomations in the continent according to existing epidemiological data. The antivenom is currently under production and has already been employed in the field in a pragmatic Phase III clinical trial in the Republic of Benin. It is a purified lyophilized polyvalent equine F(ab')(2)-based product obtained by immunization with the venoms of eleven species of African snakes of the Genera Echis, Bitis, Naja and Dendroaspis. The criteria for its design are discussed, particularly in terms of the implementation of realistic public health policies targeting mostly rural populations in the continent.


Asunto(s)
Antivenenos/inmunología , Venenos Elapídicos/antagonistas & inhibidores , Venenos de Víboras/antagonistas & inhibidores , África , Animales , Antivenenos/biosíntesis , Antivenenos/química , Trastornos de la Coagulación Sanguínea , Cromatografía Líquida de Alta Presión , Estabilidad de Medicamentos , Venenos Elapídicos/inmunología , Venenos Elapídicos/toxicidad , Hemorragia , Caballos , Dosificación Letal Mediana , Ratones , Necrosis , Pruebas de Neutralización , Especificidad de la Especie , Venenos de Víboras/inmunología , Venenos de Víboras/toxicidad
14.
Toxicon ; 48(1): 64-74, 2006 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-16759681

RESUMEN

We report the cloning of sphingomyelinase D (SMD) cDNA from Loxosceles reclusa, Loxosceles boneti and Loxosceles laeta into bacterial expression systems, as well as optimization of expression conditions so as to obtain soluble and active recombinant enzymes. The recombinant mature SMDs, tagged with a histidine tail at the N- or C-termini, were compared in terms of toxicity and enzymatic activity, and were used as immunogens for the production of monovalent antisera in rabbits and F(ab')(2) preparations in animals used for commercial antivenom production (horses). We performed studies on in vitro inhibition of enzymatic activity of natural venom preparations by antibodies generated against the tagged proteins. We also present and discuss the results of studies on the specific and para-specific in vivo protective potential of the rabbit and equine antibody preparations against the recombinant proteins themselves and natural venom preparations. Our conclusions support the feasibility of using recombinant SMDs for production and evaluation of polyvalent anti-Loxosceles antivenoms, and we offer data on the potential of paraspecific neutralization in the context of the antigenic groupings and the molecular phylogeny of those active SMDs for which amino acid sequence information is available.


Asunto(s)
Antivenenos/inmunología , Hidrolasas Diéster Fosfóricas/inmunología , Venenos de Araña/inmunología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Reacciones Cruzadas , Fragmentos Fab de Inmunoglobulinas/inmunología , Datos de Secuencia Molecular , Hidrolasas Diéster Fosfóricas/química , Conejos , Proteínas Recombinantes/inmunología
15.
Gac Med Mex ; 141(3): 215-22, 2005.
Artículo en Español | MEDLINE | ID: mdl-16025987

RESUMEN

Among the human pathologies produced by venomous animals, bee stings constitute the largest number of accidents in several countries, exceeding the mortality rate caused by other venomous animals such as snakes, spiders or scorpions. The clinical picture after the bee sting may include anaphylaxis or poisoning. The latter is produced by massive attacks and is a serious problem that may put the patient's life at risk. People that are poisoned display hemolysis, rhabdomiolysis and acute renal failure that together with other systemic failures can bring about death. The knowledge of the physiopathological mechanisms involved in the massive attack of bees is crucial for health care professionals as to date we do not have antivenoms with proven clinical efficacy. In this review we include the bee's biological aspects, venom composition and its relation with the occurrence and severity of accidents as well as epidemiological data that can be useful for this type of accidents.


Asunto(s)
Abejas , Mordeduras y Picaduras de Insectos , Animales , Venenos de Abeja/química , Venenos de Abeja/farmacología , Humanos , Mordeduras y Picaduras de Insectos/diagnóstico , Mordeduras y Picaduras de Insectos/terapia
16.
Gac. méd. Méx ; Gac. méd. Méx;141(3): 215-222, may.-jun. 2005. ilus
Artículo en Español | LILACS | ID: lil-632123

RESUMEN

Dentro de las patologías humanas producidas por animales con la capacidad de inocular veneno, las picaduras de abeja producen el mayor número de accidentes por animales en muchos países, superando a menudo en mortalidad a los producidos por serpientes, escorpiones y arañas. El cuadro clínico por la picadura de estos himenópteros puede consistir en fenómenos alérgicos o en cuadros de envenenamiento. Estos últimos se producen por el ataque de enjambres constituyendo un hecho grave que puede comprometer la vida. En el sujeto envenenado pueden observarse hemólisis, rabdomiólisis e insuficiencia renal, que junto a otras alteraciones sistémicas pueden conducir a la muerte. El conocimiento de los acontecimientos fisiopatológicos que se producen ante los ataques masivos de abejas es de suma importancia para el personal de salud dado que hasta la fecha no existen antivenenos que hayan demostrado tener eficacia clínica comprobada. En esta revisión se consideran los aspectos biológicos de las abejas y de la composición de su veneno relacionado con la ocurrencia y severidad de los accidentes, así como datos epidemiológicos de utilidad para enfrentarse a este tipo de cuadro.


Among the human pathologies produced by venomous animals, bee stings constitute the largest number of accidents in several countries, exceeding the mortality rate caused by other venomous animals such as snakes, spiders or scorpions. The clinical picture after the bee sting may include anaphylaxis or poisoning. The latter is produced by massive attacks and is a serious problem that may put the patient's life at risk. People that are poisoned display hemolysis, rhabdomiolysis and acute renal failure that together with other systemic failures can bring about death. The knowledge of the physiopathological mechanisms involved in the massive attack of bees is crucial for health care professionals as to date we do not ha ve antivenoms with proven clinical efficacy. In this review we include the bee's biological aspects, venom composition and its relation with the occurrence and severity of accidents as well as epidemiological data that can be useful for this type of accidents.


Asunto(s)
Animales , Humanos , Abejas , Mordeduras y Picaduras de Insectos , Venenos de Abeja/química , Venenos de Abeja/farmacología , Mordeduras y Picaduras de Insectos/diagnóstico , Mordeduras y Picaduras de Insectos/terapia
17.
Gac Med Mex ; 141(1): 13-21, 2005.
Artículo en Español | MEDLINE | ID: mdl-15754746

RESUMEN

The characterization of the toxic activities of snake venoms is necessary to understand the physiopathology of the envenomation and to test the potency of the antivenoms used to treat this pathology. Because of the lack of data on the toxic activities of venoms from Mexican snakes of medical importance, we studied the venoms from Bothrops asper, Athropoides nummifrr, Agkistrodon billineatus, Crotalus durissus durissus, Crotalus basiliscus, Crotalus scutulatus, Crotalus atrox and Micrurus nigrocinctus. The studies performed were: SDS-PAOE, determination of lethal potency, hemorrhagic, necrotizing, coagulation on plasma and fibrinogen, phospholipasic and fibri(noge)nolytic activities. In addition we studied the neutralizing capacity of the toxic activities of an antivenom currently used for the treatment of snakebites in Mexico. The venom from viperids showed important hemorrhagic, necrotizing, coagulative on plasma, prothrombinic, fibrinogenolytic and phospholipase activities. The venoms with the highest lethal potency were those of Micrurus nigrocinctus and Crotalus scutulatus; however, the viperine venom that globally displayed the most potent toxic activities was from Bothrops asper. All the venoms showed toxic activities of similar range to those described for other American venomous snakes. The activity on plasma or fibrinogen varied widely among the different venoms but all displayed capacity to act on the coagulation system. The antivenom tested not only neutralized the lethality B. asper venom but also its other toxic activities.


Asunto(s)
Venenos de Serpiente/toxicidad , Animales , México , Ratones
18.
Gac. méd. Méx ; Gac. méd. Méx;141(1): 13-21, ene.-feb. 2005. ilus, tab
Artículo en Español | LILACS | ID: lil-632105

RESUMEN

La caracterización de las actividades tóxicas de los venenos de serpientes es necesaria para el cabal entendimiento de los procesos fisiopatológicos que se producen ante su mordedura, como también para evaluar la potencia neutralizante de los antivenenos utilizados para tratar estos envenenamientos. A causa de los pocos datos disponibles sobre la toxicidad del veneno de serpientes con importancia sanitaria en México, estudiamos las actividades tóxicas de los venenos de Bothrops asper, Athropoides nummifer, Agkistrodon billineatus> Crotalus durissus durissus, Crotalus basiliscus, Crotalus scutulatus, Crotalus atrox y Micrurus nigrocinctus. A los venenos se les realizaron los siguientes estudios: SDS-PAUE, determinación de la potencia letal, y de las actividades hemorrágica, necrotizante, coagulante en plasma y fibrinógeno, fosfolipásica y fibrinogenolítica. Se estudió además la capacidad neutralizante de un antiveneno de uso corriente para la terapéutica de las mordeduras de serpientes venenosas en México, sobre varias de estas actividades. Los venenos de vipéridos mostraron actividades hemorrágicas, necrotizante, coagulante sobre plasma, protrombínica, fibrinogenolítica y fosfolipásica importantes. Los venenos de mayor potencia letal fueron los de Micrurus nigrocinctus y Crotalus scutulatus, sin embargo el veneno que presentó en general potencias tóxicas mayores fue el de Bothrops asper. Las diferentes potencias tóxicas halladas se encontraron dentro de los márgenes descritos para especies de vipéridos y elápidos de Sudamérica. La actividad sobre el plasma y el fibrinógeno fue muy diferente en los diferentes venenos viperinos, sin embargo todos mostraron ser capaces de afectar componentes del sistema de la coagulación. El antiveneno probado no sólo neutralizó la letalidad del veneno sino también sus actividades tóxicas.


The characterization of the toxic activities of snake venoms is necessary to understand the physiopathology of the envenomation and to test the potency of the antivenoms used to treat this pathology. Because of the lack of data on the toxic activities of venoms from Mexican snakes of medical importance, we studied the venoms from Bothrops asper, Athropoides nummifer, Agkistrodon billineatus, Crotalus durissus durissus, Crotalus basiliscus, Crotalus scutulatus, Crotalus atrox and Micrurus nigrocinctus. The studies performed were : SDS-PAOE, determination of lethal potency, hemorrhagic, necrotizing, coagulation on plasma and fibrinogen, phospholipasic and fibrinogenolytic activities. In addition we studied the neutralizing capacity of the toxic activities of an antivenom currently used for the treatment of snakebites in Mexico. The venom from viperids showed important hemorrhagic, necrotizing, coagulative on plasma, prothrombinic, fibrinogenolytic and phospholipase activities. The venoms with the highest lethal potency were those of Micrurus nigrocinctus and Crotalus scutulatus; however, the viperine venom that globally displayed the most potent toxic activities was from Bothrops asper. All the venoms showed toxic activities of similar range to those described for other American venomous snakes. The activity on plasma or fibrinogen varied widely among the different venoms but all displayed capacity to act on the coagulation system. The antivenom tested not only neutralized the lethalityB. asper venom but also its other toxic activities.


Asunto(s)
Animales , Ratones , Venenos de Serpiente/toxicidad , México
19.
Toxicon ; 44(5): 507-14, 2004 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-15450925

RESUMEN

In this study we report the isolation and characterization of several sphingomyelinase D isoforms from the venoms of the North American spiders Loxosceles boneti and Loxosceles reclusa, from Mexico and the United States, respectively. We have measured their enzymatic activity, their capacity to induce necrotic lesions in rabbits, cloned the cDNAs coding for the mature forms of two of the isoforms from L. boneti and two of L. reclusa based on N-terminal sequence information of the purified proteins, and performed a comprehensive comparison of the sequence data generated by us with that reported for other sphingomyelinase genes to date.


Asunto(s)
Isoenzimas/genética , Hidrolasas Diéster Fosfóricas/genética , Venenos de Araña/química , Arañas/enzimología , Animales , Clonación Molecular , ADN Complementario/biosíntesis , ADN Complementario/genética , Isoenzimas/metabolismo , Datos de Secuencia Molecular , Necrosis , América del Norte , Hidrolasas Diéster Fosfóricas/aislamiento & purificación , Hidrolasas Diéster Fosfóricas/metabolismo , Conejos , Homología de Secuencia de Aminoácido , Especificidad de la Especie , Venenos de Araña/genética , Arañas/genética , Arañas/metabolismo
20.
J Toxicol Clin Toxicol ; 42(2): 171-8, 2004.
Artículo en Inglés | MEDLINE | ID: mdl-15214622

RESUMEN

Micrurus snakes (coral snakes) may produce severe envenomation that can lead to death by peripheral respiratory paralysis. Only few laboratories produce specific antivenoms, and despite the cross-reactivity found in some Micrurus species venoms, the treatment is not always effective. To test two therapeutic antivenoms against the venom of four species of Micrurus from Southern America, North of South America, Central America, and North America, the determination of the lethal potency of the venoms, the study of some biochemical and immunochemical characteristics, and the determination of the neutralizing activity of both antivenoms were studied. North American and South American antivenoms neutralized well venoms from Micrurus species of the corresponding hemisphere but displayed lower effectiveness against venoms of species from different hemispheres. It was concluded that the neutralization of Micrurus venoms by regional antivenoms could be useful to treat the envenomation by some Micrurus snakes but is necessary to evaluate carefully the antivenoms to be used with the venoms from the snakes of the region. Also, considering the difficulties for coral snake antivenom production, the development of a polyvalent antivenom is useful to treat the envenomation by coral snakes from different regions is necessary.


Asunto(s)
Antivenenos/uso terapéutico , Venenos Elapídicos/toxicidad , Elapidae , Mordeduras de Serpientes/tratamiento farmacológico , Américas , Animales , Antivenenos/inmunología , Bioensayo , Reacciones Cruzadas , Venenos Elapídicos/antagonistas & inhibidores , Venenos Elapídicos/inmunología , Inyecciones Intraperitoneales , Dosificación Letal Mediana , Ratones , Pruebas de Neutralización , Mordeduras de Serpientes/inmunología , Especificidad de la Especie
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