RESUMEN
Snakebite accidents are a public health problem that affects the whole world, causing thousands of deaths and amputations each year. In Brazil, snakebite envenomations are caused mostly by snakes from the Bothrops genus. The local symptoms are characterized by pain, swelling, ecchymosis, and hemorrhages. Systemic disturbances can lead to necrosis and amputations. The present treatment consists of intravenous administration of bothropic antivenom, which is capable of reversing most of the systemic symptoms, while presenting limitations to treat the local effects, such as hemorrhage and to neutralize the snake venom serine protease (SVSP). In this context, we aimed to evaluate the activity of selective serine protease inhibitors (pepC and pepB) in combination with the bothropic antivenom in vivo. Further, we assessed their possible synergistic effect in the treatment of coagulopathy and hemorrhage induced by Bothrops jararaca venom. For this, we evaluated the in vivo activity in mouse models of local hemorrhage and a series of in vitro hemostasis assays. Our results showed that pepC and pepB, when combinated with the antivenom, increase its protective activity in vivo and decrease the hemostatic disturbances in vitro with high selectivity, possibly by inhibiting botropic proteases. These data suggest that the addition of serine protease inhibitor to the antivenom can improve its overall potential.
Asunto(s)
Bothrops , Venenos de Crotálidos , Animales , Antivenenos/farmacología , Antivenenos/uso terapéutico , Brasil , Venenos de Crotálidos/toxicidad , Hemorragia/inducido químicamente , Hemorragia/tratamiento farmacológico , Ratones , Inhibidores de Serina Proteinasa/farmacología , Inhibidores de Serina Proteinasa/uso terapéuticoRESUMEN
BACKGROUND: Transplant renal artery stenosis (TRAS), the most common vascular complication after transplant (Tx), leads to resistant hypertension, impaired renal function, and even loss of the graft. The purpose of the study was to investigate the prevalence and factors associated with TRAS in northeastern Brazil. METHODS: The study was conducted as a retrospective case-control study in a population of Tx recipients in a renal Tx center in northeastern Brazil. Demographic and clinical characteristics of the recipients and donors, data related to the surgery, laboratory data, and number of anti-hypertensive drugs were assessed. Statistical analysis was performed with the use of SPSS 17.0. RESULTS: A total of 494 of 529 recipients were assessed, of which 24 had TRAS. The prevalence of TRAS was 4.8%. Twelve patients (50%) were men with a mean age of 46.7 ± 13.5 years. The mean time of diagnosis was 89.9 days after Tx. The risk factors associated with TRAS were number of anti-hypertensive drugs ≥2 (odds ratio, 17.0; confidence interval, 4.1 to 70.4; P = .001) and grafting with 2 or more arteries (odds ratio, 8.9; confidence interval, 1.4 to 56.6; P = .021). There was a significant reduction in mean systolic blood pressure (147.1 ± 23.7 to 127.8 ± 15.2 mm Hg, P = .001) and diastolic blood pressure (86.6 ± 13.0 to 77.6 ± 9.4 mm Hg, P = .001) after TRAS repair and in serum creatinine (2.8 ± 2.4 to 1.9 ± 1.8 mg/dL, P = .04). CONCLUSIONS: Grafts with 2 or more arteries are associated with TRAS, as well as patients who use a higher number of anti-hypertensive drugs. TRAS repair was associated with improved blood pressure control and renal function.
Asunto(s)
Oclusión de Injerto Vascular/etiología , Trasplante de Riñón/efectos adversos , Obstrucción de la Arteria Renal/etiología , Adulto , Antihipertensivos/uso terapéutico , Presión Sanguínea , Brasil/epidemiología , Estudios de Casos y Controles , Femenino , Oclusión de Injerto Vascular/epidemiología , Oclusión de Injerto Vascular/fisiopatología , Humanos , Riñón/irrigación sanguínea , Riñón/cirugía , Trasplante de Riñón/métodos , Masculino , Persona de Mediana Edad , Prevalencia , Arteria Renal/fisiopatología , Obstrucción de la Arteria Renal/epidemiología , Obstrucción de la Arteria Renal/fisiopatología , Insuficiencia Renal/etiología , Estudios Retrospectivos , Factores de Riesgo , Resultado del TratamientoRESUMEN
Hambúrgueres foram elaborados manualmente com carne mecanicamente separada (CMS) de tilápia, em um planejamento fatorial 22 completo, visando determinar o efeito da fibra de trigo e óleo de milho no encolhimento e na capacidade de retenção de água. A melhor formulação obtida foi a interação de 1% de óleo de milho e 2,5% de fibra de trigo e como resposta 4,3% de encolhimento e 87% de capacidade de retenção de água. Na segunda etapa, a melhor formulação de CMS foi elaborada manualmente em quatro bateladas e submetidas às análises microbiológicas, físico-químicas e sensorial, em triplicata. Os hambúrgueres encontravam-se dentro dos padrões permitidos pela legislação brasileira. A fibra de trigo e o óleo vegetal de milho podem ser utilizados na elaboração de fishbúrguer de CMS de tilápia, com excelente aceitação sensorial.
RESUMEN
O uso do soro lácteo como complemento na formulação de alimentos visando ao seu aproveitamento é uma alternativa para a obtenção de novos produtos. Neste trabalho, desenvolveram-se bebidas à base de soro oriundo de queijo coalho tipo A com frutas e hortaliças. Na formulação das bebidas, utilizaram-se diferentes proporções entre soro lácteo, frutas e hortaliças, com adição de 10% de açúcar. Seis bebidas foram formuladas e submetidas a um teste de preferência, e as três primeiras colocadas foram submetidas a um teste de aceitação sensorial. As três preferidas foram as bebidas sabor graviola, morango e goiaba. A bebida sabor graviola obteve aceitação com as melhores notas nos atributos sensoriais cor, sabor, aparência e qualidade global, seguida pela formulação de sabor morango, mas sem diferença significativa (P>0,05). A bebida sabor goiaba teve o diferencial no atributo odor e superou as bebidas sabor graviola e morango (P<0,05). Conclui-se que é viável a elaboração de bebidas à base de soro de queijo coalho com frutas.
The use of whey as complement in food formulation aiming it's an alternative for the attainment of new products. The aim of this work was developed a type A curd cheese whey derived drink with fruits and vegetables. On drink's formulation was used different ratios of whey, fruits and vegetables, with 10% of sugar. Six drinks were formulated and submited to a preference test. The three best drinks were submitted to a acceptance test. The three best drinks were graviola, strawberry and guava flavours. The graviola one had the greater acceptability with best grades in color's sensory attributes, flavor, appearance and overall quality, following the strawberry flavor, but without significant difference (P<0,05). The guava dink had it's differential on scent character, overcoming the graviola and strawberry drinks (P<0,05). In conclusion we can say that whey-based drink with fruits is viable.
Asunto(s)
Animales , Alimentos/clasificación , Bebidas/análisis , Frutas , Queso , Verduras , Daucus carota , MenthaRESUMEN
O uso do soro lácteo como complemento na formulação de alimentos visando ao seu aproveitamento é uma alternativa para a obtenção de novos produtos. Neste trabalho, desenvolveram-se bebidas à base de soro oriundo de queijo coalho tipo A com frutas e hortaliças. Na formulação das bebidas, utilizaram-se diferentes proporções entre soro lácteo, frutas e hortaliças, com adição de 10% de açúcar. Seis bebidas foram formuladas e submetidas a um teste de preferência, e as três primeiras colocadas foram submetidas a um teste de aceitação sensorial. As três preferidas foram as bebidas sabor graviola, morango e goiaba. A bebida sabor graviola obteve aceitação com as melhores notas nos atributos sensoriais cor, sabor, aparência e qualidade global, seguida pela formulação de sabor morango, mas sem diferença significativa (P>0,05). A bebida sabor goiaba teve o diferencial no atributo odor e superou as bebidas sabor graviola e morango (P<0,05). Conclui-se que é viável a elaboração de bebidas à base de soro de queijo coalho com frutas.(AU)
The use of whey as complement in food formulation aiming it's an alternative for the attainment of new products. The aim of this work was developed a type A curd cheese whey derived drink with fruits and vegetables. On drink's formulation was used different ratios of whey, fruits and vegetables, with 10% of sugar. Six drinks were formulated and submited to a preference test. The three best drinks were submitted to a acceptance test. The three best drinks were graviola, strawberry and guava flavours. The graviola one had the greater acceptability with best grades in color's sensory attributes, flavor, appearance and overall quality, following the strawberry flavor, but without significant difference (P<0,05). The guava dink had it's differential on scent character, overcoming the graviola and strawberry drinks (P<0,05). In conclusion we can say that whey-based drink with fruits is viable.(AU)
Asunto(s)
Animales , Alimentos/clasificación , Bebidas/análisis , Queso , Frutas , Verduras , Mentha , Daucus carotaRESUMEN
Lopap, found in the bristles of Lonomia obliqua caterpillar, is the first exogenous prothrombin activator that shows serine protease-like activity, independent of prothrombinase components and unique lipocalin reported to interfere with hemostasis mechanisms. To assess the action of an exogenous prothrombin activator reversing the anticoagulant and antihemostatic effect induced by low molecular weight heparin (LMWH), male New Zealand rabbits (N = 20, weighing 3.8-4.0 kg) allocated to 4 groups were anticoagulated with 1800 IU/kg LMWH (iv) over 2 min, followed by iv administration of saline (SG) or recombinant Lopap (rLopap) at 1 µg/kg (LG1) or 10 µg/kg (LG10), 10 min after the injection of LMWH, in a blind manner. Control animals (CG) were treated only with saline. The action of rLopap was assessed in terms of activated partial thromboplastin time (aPTT), prothrombin fragment F1+2, fibrinogen, and ear puncture bleeding time (BT) at 5, 10, 15, 17, 20, 30, 40, 60, and 90 min after initiation of LMWH infusion. LG10 animals showed a decrease of aPTT in more than 50% and BT near to normal baseline. The level of prothrombin fragment F1+2 measured by ELISA had a 6-fold increase with rLopap treatment (10 µg/kg) and was inversely proportional to BT in LMWH-treated animals. Thus, Lopap, obtained in recombinant form using E. coli expression system, was useful in antagonizing the effect of LMWH through direct prothrombin activation, which can be a possible strategy for the reversal of bleeding and anticoagulant events.
Asunto(s)
Animales , Masculino , Conejos , Anticoagulantes/uso terapéutico , Coagulación Sanguínea/efectos de los fármacos , Hemorragia/tratamiento farmacológico , Antagonistas de Heparina/farmacología , Heparina de Bajo-Peso-Molecular/antagonistas & inhibidores , Protrombina/efectos de los fármacos , Serina Endopeptidasas/farmacología , Hemorragia/inducido químicamenteRESUMEN
Lopap, found in the bristles of Lonomia obliqua caterpillar, is the first exogenous prothrombin activator that shows serine protease-like activity, independent of prothrombinase components and unique lipocalin reported to interfere with hemostasis mechanisms. To assess the action of an exogenous prothrombin activator reversing the anticoagulant and antihemostatic effect induced by low molecular weight heparin (LMWH), male New Zealand rabbits (N = 20, weighing 3.8-4.0 kg) allocated to 4 groups were anticoagulated with 1800 IU/kg LMWH (iv) over 2 min, followed by iv administration of saline (SG) or recombinant Lopap (rLopap) at 1 µg/kg (LG1) or 10 µg/kg (LG10), 10 min after the injection of LMWH, in a blind manner. Control animals (CG) were treated only with saline. The action of rLopap was assessed in terms of activated partial thromboplastin time (aPTT), prothrombin fragment F1+2, fibrinogen, and ear puncture bleeding time (BT) at 5, 10, 15, 17, 20, 30, 40, 60, and 90 min after initiation of LMWH infusion. LG10 animals showed a decrease of aPTT in more than 50% and BT near to normal baseline. The level of prothrombin fragment F1+2 measured by ELISA had a 6-fold increase with rLopap treatment (10 µg/kg) and was inversely proportional to BT in LMWH-treated animals. Thus, Lopap, obtained in recombinant form using E. coli expression system, was useful in antagonizing the effect of LMWH through direct prothrombin activation, which can be a possible strategy for the reversal of bleeding and anticoagulant events.
Asunto(s)
Anticoagulantes/uso terapéutico , Coagulación Sanguínea/efectos de los fármacos , Hemorragia/tratamiento farmacológico , Antagonistas de Heparina/farmacología , Heparina de Bajo-Peso-Molecular/antagonistas & inhibidores , Protrombina/efectos de los fármacos , Serina Endopeptidasas/farmacología , Animales , Hemorragia/inducido químicamente , Masculino , ConejosRESUMEN
Studies on mycorrhizal symbiosis effects on metal accumulation and plant tolerance are not common in perennial crops under metal stress. The objective of this study was to evaluate the influence of mycorrhization on coffee seedlings under Cu and Zn stress. Copper (Cu) and zinc (Zn) uptake and some biochemical and physiological traits were studied in thirty-week old Coffea arabica seedlings, in response to the inoculation with arbuscular mycorrhizal fungi (AMF) and to increasing concentrations of Cu or Zn in soil. The experiments were conducted under greenhouse conditions in a 2×4 factorial design (inoculation or not with AMF and 0, 50, 150 and 450mgkg(-1) Cu or 0, 100, 300 and 900mgkg(-1) Zn). Non-mycorrhizal plants maintained a hampered and slow growth even in a soil with appropriate phosphorus (P) levels for this crop. As metal levels increased in soil, a greater proportion of the total absorbed metals were retained by roots. Foliar Cu concentrations increased only in non-mycorrhizal plants, reaching a maximum concentration of 30mgkg(-1) at the highest Cu in soil. Mycorrhization prevented the accumulation of Cu in leaves, and mycorrhizal plants showed higher Cu contents in stems, which indicated a differential Cu distribution in AMF-associated or non-associated plants. Zn distribution and concentrations in different plant organs followed a similar pattern independently of mycorrhization. In mycorrhizal plants, only the highest metal concentrations caused a reduction in biomass, leading to significant changes in some biochemical indicators, such as malondialdehyde, proline and amino acid contents in leaves and also in foliar free amino acid composition. Marked differences in these physiological traits were also found due to mycorrhization. In conclusion, AMF protected coffee seedlings against metal toxicity.
Asunto(s)
Coffea/metabolismo , Cobre/metabolismo , Micorrizas/metabolismo , Contaminantes del Suelo/metabolismo , Simbiosis , Zinc/metabolismo , Aminoácidos/metabolismo , Coffea/efectos de los fármacos , Coffea/microbiología , Cobre/toxicidad , Productos Agrícolas/efectos de los fármacos , Productos Agrícolas/metabolismo , Productos Agrícolas/microbiología , Relación Dosis-Respuesta a Droga , Peroxidación de Lípido/efectos de los fármacos , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/metabolismo , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/metabolismo , Raíces de Plantas/microbiología , Plantones/efectos de los fármacos , Plantones/metabolismo , Contaminantes del Suelo/toxicidad , Estrés Fisiológico , Zinc/toxicidadRESUMEN
Realizaram-se avaliações bacteriológicas da matéria-prima utilizada na elaboração de um produto cárneo caprino tipo hambúrguer defumado, antes e após sanitização. A carne foi tratada por aspersão, com uma solução de ácidos orgânicos contendo 2 por cento de ácido acético, 1 por cento de ácido lático, 0,25 por cento de ácido cítrico e 0,1 por cento de ácido ascórbico, e armazenada sob refrigeração. As análises bacteriológicas foram realizadas no dia 0 e aos 7 e 14 dias após defumação. Na carne, no dia 0, a contagem de coliformes fecais estava acima do limite permitido pela legislação vigente, mas sete dias após a sanitização, esse parâmetro estava dentro dos limites permitidos. No produto final todos os parâmetros bacteriológicos encontraram-se dentro dos padrões higiênico-sanitários.
In order to obtain a safe goat smoked hamburger, bacteriological analysis of the raw material was performed before and after sanitation. Meat was sprayed with a solution of organic acids containing 2 percent acetic acid, 1 percent lactic acid, 0.25 percent citric acid and 0.1 percent ascorbic acid and, after that, stored under refrigeration. Bacteriological analysis was carried out on days 0, 7 and 14 after goat smoked hamburger processing. In meat, on day 0, the fecal coliform count was above legal limits, but on day 7 after sanitation this parameter was these limits. In the final product, all the results of the bacteriological parameters were according to the sanitary-hygienic standards.
Asunto(s)
Técnicas Bacteriológicas , Carne/análisis , CabrasRESUMEN
Realizaram-se avaliações bacteriológicas da matéria-prima utilizada na elaboração de um produto cárneo caprino tipo hambúrguer defumado, antes e após sanitização. A carne foi tratada por aspersão, com uma solução de ácidos orgânicos contendo 2% de ácido acético, 1% de ácido lático, 0,25% de ácido cítrico e 0,1% de ácido ascórbico, e armazenada sob refrigeração. As análises bacteriológicas foram realizadas no dia 0 e aos 7 e 14 dias após defumação. Na carne, no dia 0, a contagem de coliformes fecais estava acima do limite permitido pela legislação vigente, mas sete dias após a sanitização, esse parâmetro estava dentro dos limites permitidos. No produto final todos os parâmetros bacteriológicos encontraram-se dentro dos padrões higiênico-sanitários.(AU)
In order to obtain a safe goat smoked hamburger, bacteriological analysis of the raw material was performed before and after sanitation. Meat was sprayed with a solution of organic acids containing 2% acetic acid, 1% lactic acid, 0.25% citric acid and 0.1% ascorbic acid and, after that, stored under refrigeration. Bacteriological analysis was carried out on days 0, 7 and 14 after goat smoked hamburger processing. In meat, on day 0, the fecal coliform count was above legal limits, but on day 7 after sanitation this parameter was these limits. In the final product, all the results of the bacteriological parameters were according to the sanitary-hygienic standards.(AU)
Asunto(s)
Técnicas Bacteriológicas , Carne/análisis , CabrasRESUMEN
SMase I, a 32 kDa sphingomyelinase found in Loxosceles laeta venom, is responsible for the major pathological effects of spider envenomation. This toxin has been cloned and functionally expressed as a fusion protein containing a 6 x His tag at its N-terminus to yield a 33 kDa protein [Fernandes-Pedrosa et al. (2002), Biochem. Biophys. Res. Commun. 298, 638-645]. The recombinant protein possesses all the biological properties ascribed to the whole L. laeta venom, including dermonecrotic and complement-dependent haemolytic activities. Dynamic light-scattering experiments conducted at 291 K demonstrate that the sample possesses a monomodal distribution, with a hydrodynamic radius of 3.57 nm. L. laeta SMase I was crystallized by the hanging-drop vapour-diffusion technique using the sparse-matrix method. Single crystals were obtained using a buffer solution consisting of 0.08 M HEPES and 0.9 M trisodium citrate, which was titrated to pH 7.5 using 0.25 M sodium hydroxide. Complete three-dimensional diffraction data were collected to 1.8 angstroms at the Laboratório Nacional de Luz Síncrotron (LNLS, Campinas, Brazil). The crystals belong to the hexagonal system (space group P6(1) or P6(5)), with unit-cell parameters a = b = 140.6, c = 113.6 angstroms. A search for heavy-atom derivatives has been initiated and elucidation of the crystal structure is currently in progress.
Asunto(s)
Esfingomielina Fosfodiesterasa/química , Venenos de Araña/química , Animales , Citratos/farmacología , Cristalografía por Rayos X , ADN Complementario/metabolismo , Luz , Unión Proteica , Conformación Proteica , Estructura Terciaria de Proteína , Proteínas Recombinantes/química , Dispersión de Radiación , Citrato de Sodio , Arañas , TemperaturaRESUMEN
The specific Kunitz Bauhinia ungulata factor Xa inhibitor (BuXI) and the Bauhinia variegata trypsin inhibitor (BvTI) blocked the activity of trypsin, chymotrypsin, plasmin, plasma kallikrein and factor XIIa, and factor Xa inhibition was achieved only by BuXI (K(i) 14 nM). BuXI and BvTI are highly homologous (70%). The major differences are the methionine residues at BuXI reactive site, which are involved in the inhibition, since the oxidized protein no longer inhibits factor Xa but maintains the trypsin inhibition. Quenched fluorescent substrates based on the reactive site sequence of the inhibitors were synthesized and the kinetic parameters of the hydrolysis were determined using factor Xa and trypsin. The catalytic efficiency k(cat)/K(m) 4.3 x 10(7) M(-1)sec(>-1) for Abz-VMIAALPRTMFIQ-EDDnp (lead peptide) hydrolysis by factor Xa was 10(4)-fold higher than that of Boc-Ile-Glu-Gly-Arg-AMC, widely used as factor Xa substrate. Lengthening of the substrate changed its susceptibility to factor Xa hydrolysis. Both methionine residues in the substrate influence the binding to factor Xa. Serine replacement of threonine (P(1)') decreases the catalytic efficiency by four orders of magnitude. Factor Xa did not hydrolyze the substrate containing the reactive site sequence of BvTI, that inhibits trypsin inhibitor but not factor Xa. Abz-VMIAALPRTMFIQ-EDDnp prolonged both the prothrombin time and the activated partial thromboplastin time, and the other modified substrates used in this experiment altered blood-clotting assays.
Asunto(s)
Bauhinia/química , Inhibidores del Factor Xa , Proteínas de Plantas/metabolismo , Inhibidores de Serina Proteinasa/metabolismo , Secuencia de Aminoácidos , Animales , Sitios de Unión , Bovinos , Factor Xa/química , Colorantes Fluorescentes , Humanos , Cinética , Datos de Secuencia Molecular , Proteínas de Plantas/aislamiento & purificación , Semillas/química , Homología de Secuencia , Inhibidores de Serina Proteinasa/aislamiento & purificación , Especificidad por SustratoRESUMEN
Lonomia obliqua venom causes a severe consumptive coagulopathy, which can lead to a hemorrhagic syndrome. The crude bristles extract displays a procoagulant activity due to a Factor X and to a prothrombin activating activity. Here, we describe a 69 kDa prothrombin activator serine protease purified from L. obliqua caterpillar bristle extract using gel filtration (Sephadex G 75) and HPLC (C(4) column). The purified protein was able to activate prothrombin in a dose-dependent manner, and calcium ions increased this activity. The prothrombin-derived fluorogenic peptide (Abz-YQTFFNPRTGSQ-EDDnp) had its main cleavage site at the Arg-Thr bond. The kinetic parameters obtained for this substrate were Kmapp of 4.5 microM, kcat of 5.32 s(-1), and a kcat/Kmapp of 1.2 x 10(6) M(-1) s(-1). The prothrombin fragments generated by the purified enzyme corresponded to the molecular masses of prethrombin 2, fragment 1, fragment 2, and thrombin as seen in SDS-PAGE. The thrombin generated was able to clot purified fibrinogen. The partial amino acid sequence of the purified protein, named Lopap (L. obliqua prothrombin activator protease), showed no similarity to any known prothrombin activator.
Asunto(s)
Protrombina/metabolismo , Serina Endopeptidasas/farmacología , Secuencia de Aminoácidos , Animales , Venenos de Artrópodos/aislamiento & purificación , Venenos de Artrópodos/farmacología , Sitios de Unión , Pruebas de Coagulación Sanguínea , Calcio/farmacología , Relación Dosis-Respuesta a Droga , Factor X/metabolismo , Fibrinógeno/efectos de los fármacos , Fibrinógeno/metabolismo , Colorantes Fluorescentes/metabolismo , Humanos , Cinética , Larva , Lepidópteros , Datos de Secuencia Molecular , Serina Endopeptidasas/aislamiento & purificaciónRESUMEN
We have previously described Kunitz-type serine proteinase inhibitors purified from Bauhinia seeds. Human plasma kallikrein shows different susceptibility to those inhibitors. In this communication, we describe the interaction of human plasma kallikrein with fluorogenic and non-fluorogenic peptides based on the Bauhinia inhibitors' reactive site. The hydrolysis of the substrate based on the B. variegata inhibitor reactive site sequence, Abz-VVISALPRSVFIQ-EDDnp (Km 1.42 microM, kcat 0.06 s(-1), and kcat/Km 4.23 x 10(4) M(-1) s(-1)), is more favorable than that of Abz-VMIAALPRTMFIQ-EDDnp, related to the B. ungulata sequence (Km 0.43 microM, kcat 0.00017 s(-1), and kcat/Km 3.9 x 10(2) M(-1) s(-1)). Human plasma kallikrein does not hydrolyze the substrates Abz-RPGLPVRFESPL-EDDnp and Abz-FESPLRINIIKE-EDDnp based on the B. bauhinioides inhibitor reactive site sequence, the most effective inhibitor of the enzyme. These peptides are competitive inhibitors with Ki values in the nM range. The synthetic peptide containing 19 amino acids based on the B. bauhinioides inhibitor reactive site (RPGLPVRFESPL) is poorly cleaved by kallikrein. The given substrates are highly specific for trypsin and chymotrypsin hydrolysis. Other serine proteinases such as factor Xa, factor XII, thrombin and plasmin do not hydrolyze B. bauhinioides inhibitor related substrates.
Asunto(s)
Colorantes Fluorescentes/farmacología , Calicreínas/metabolismo , Péptidos/farmacología , Plantas/química , Inhibidor de la Tripsina de Soja de Kunitz/farmacología , Secuencia de Aminoácidos , Animales , Sitios de Unión/efectos de los fármacos , Colorantes Fluorescentes/síntesis química , Humanos , Hidrólisis , Calicreínas/efectos de los fármacos , Datos de Secuencia Molecular , Péptidos/síntesis química , Porcinos , Inhibidor de la Tripsina de Soja de Kunitz/aislamiento & purificaciónRESUMEN
Kunitz type Bauhinia ungulata factor Xa inhibitor (BuXI) was purified from B. ungulata seeds. BuXI inactivates factor Xa and human plasma kallikrein (HuPK) with Ki values of 18.4 and 6.9 nM, respectively. However, Bauhinia variegata trypsin inhibitor (BvTI) which is 70% homologous to BuXI does not inhibit factor Xa and is less efficient on HuPK (Ki = 80 nM). The comparison between BuXI and BvTI reactive site structure indicates differences at Met59, Thr66 and Met67 residues. The hydrolysis rate of quenched fluorescence peptide substrates based on BuXI reactive site sequence, Abz-VMIAALPRTMFIQ-EDDnp (leading peptide), by HuPK and porcine pancreatic kallikrein (PoPK) is low, but hydrolysis is enhanced with Abz-VMIAALPRTMQ-EDDnp, derived from the leading peptide shortened by removing the dipeptide Phe-Ileu from the C-terminal portion, for HuPK (Km = 0.68 microM, k(cat)/Km = 1.3 x 10(6) M(-1) s(-1)), and the shorter substrate Abz-LPRTMQ-EDDnp is better for PoPK (Km = 0.66 microM, k(cat)/Km = 2.2 x 10(3) M(-1) s(-1)). The contribution of substrate methionine residues to HuPK and PoPK hydrolysis differs from that observed with factor Xa. The determined Km and k(cat) values suggest that the substrates interact with kallikreins the same as an enzyme and inhibitor interacts to form complexes.
Asunto(s)
Fabaceae/química , Inhibidores del Factor Xa , Calicreínas/sangre , Calicreínas/metabolismo , Proteínas de Plantas/aislamiento & purificación , Plantas Medicinales , Calicreínas de Tejido/metabolismo , Secuencia de Aminoácidos , Sitios de Unión , Factor Xa/metabolismo , Colorantes Fluorescentes/metabolismo , Humanos , Datos de Secuencia Molecular , Proteínas de Plantas/metabolismo , Semillas/enzimología , Semillas/metabolismo , Inhibidores de Serina Proteinasa/metabolismo , Especificidad por SustratoRESUMEN
Potential pathogenesis from pollution depends on the kind of contaminant, its concentration, its duration in environment and on host's characteristics. Faced to the threat of medical consequences of pollution, it is necessary to be alert to recognize them, with an anticipated description. Two syndromes can be distinguished, one acute and the other chronic, that may be useful as reference for an eventual epidemiologic surveillance.
Asunto(s)
Contaminación Ambiental/prevención & control , Medicina Interna , Enfermedad Aguda , Contaminantes Atmosféricos/efectos adversos , Enfermedad Crónica , Salud Ambiental , Contaminación Ambiental/efectos adversos , Humanos , México , SíndromeRESUMEN
The amount of neuroendocrine granules in microgranular cells of salivary glands were investigated in necropsies of 20 non-insulin dependent diabetics (NIDDM) and in 20 non-diabetic subjects with Grimelius and Fontana-Masson stainings. Granules in serous acini, and intercalated ducts were observed in both groups; however, a significant higher number of granules and microgranular cells were observed in NIDDM subjects as compared with non-diabetics. Both parotid and submaxillary glands were significantly heavier in NIDDM than in the non-diabetic group. These granules may be related to immunoreactive glucagon which has been found in submaxillary glands of rodents and might play a role in the pathogenesis of NIDDM. Further investigations should be performed to clarify whether these cells are the site of glucagon synthesis and also clarify the pathogenesis of NIDDM.
Asunto(s)
Gránulos Citoplasmáticos/patología , Diabetes Mellitus Tipo 2/patología , Glándula Parótida/patología , Glándula Submandibular/patología , Adulto , Anciano , Cadáver , Femenino , Humanos , Masculino , Persona de Mediana Edad , Sistemas Neurosecretores/patología , Tamaño de los ÓrganosRESUMEN
Case report of Burkitt's type lymphoma with involvement of the spinal cord in a three years-old child. An epidural tumoral mass (T11-L1) was removed surgically (Fig. 2), and clinical conditions improved for a short period of time (1 and 1/2 months) after which symptoms reappeared. At this occasion a computerized tomography showed a tumoral mass occupying the spinal canal with high density at the L2 level. The patient died 6 months after the beginning of the disease. General considerations are made on Burkitt's lymphoma.