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Objetivo: Identificar las principales preocupaciones antes y después de realizarse un procedimiento quirúrgico, e identificar diferencias basadas en género, edad y nivel de escolaridad. Material y Método: Mediante un muestreo no probabilístico incidental se aplicaron 201 encuestas semiestructuradas a personas en la sala de espera del HSJD. Las respuestas fueron procesadas mediante un análisis mixto donde se organizaron categorías por unidades de significancia y se ordenaron de forma decreciente para luego comparar según edad, género y escolaridad. Resultados: Respecto a los aspectos que las personas consideran relevante conocer previo a la cirugía, 4 categorías concentran el 71 % del total: 1. Riesgos, complicaciones y secuelas, 2. Explicación del procedimiento, 3. Beneficios y pronóstico, 4. Cuidados y evaluaciones previas. En relación con lo que le gustaría que le explicaran después, se prioriza: 1. Conocer los resultados del procedimiento (19,3%), 2. Control, derivaciones y cuidados paliativos (15%), 3. Tiempo de recuperación (13,7%). Diferencias significativas: una mayor necesidad de acompañamiento y explicación de la patología expresada por hombres, el interés por conocer y los aspectos relacionados con funcionalidad postoperatoria en personas entre 18-65 años, el interés de conocer los cuidados y evaluaciones previas, conocer los resultados del procedimiento, los cuidados de la herida y la funcionalidad posoperatoria según nivel educacional. Discusión y conclusión: Existen variaciones significativas en contenido prioritario según edad, género y nivel educacional. Finalmente, el presente artículo proporciona evidencia en cuanto a las principales prioridades de pacientes en el sistema público chileno para el consentimiento informado.
Aim: To identify the main concerns before and after undergoing a surgical procedure, and to identify differences based on gender, age and level of education. Material and Method: 201 semi-structured surveys were administered to people in the waiting room of the HSJD by means of incidental non-probabilistic sampling. The responses were processed by means of a mixed analysis in which categories were organized by units of significance and ordered in decreasing order to then compare according to age, gender and schooling. Results: Regarding the aspects that people considered relevant to know before surgery, 4 categories accounted for 71% of the total: 1. Risks, complications and sequels, 2. Explanation of the procedure, 3. Benefits and prognosis, 4. Prior care and evaluations. In relation to what you would like to have explained to you afterwards, priority is given to: 1. knowing the results of the procedure (19.3%), 2. Monitoring, referrals and palliative care (15%), 3. Recovery time (13.7%). Significant differences: a greater need for accompaniment and explanation of the pathology expressed by men, interest in knowing and aspects related to postoperative functionality in persons between 18-65 years of age, interest in knowing the previous care and evaluations, knowing the results of the procedure, wound care and postoperative functionality according to educational level. Discussion and conclusion: There are significant variations in priority content according to age, gender and educational level. Finally, the present article provides evidence regarding the main priorities of patients in the chilean public system for informed consent.
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The aims of the current study were to assess the inclusion levels of grape byproduct replacing jiggs hay in lambs diets and to evaluate signs of copper poisoning. Thirty-four Texel and Corriedale crossbred female lambs were used in a randomized block experimental design. The treatments comprised four levels of grape byproduct (0; 100; 200 and 300â¯g/kg DM) in replacement of jiggs hay. The diets were adjusted to the same ZnCu ratio (6:1) of the basal diet. Grape byproduct consumption at up to 30%, led to similar weight gain in the different treatments (Pâ¯=â¯.92), which was suitable for growing lambs. Grape byproduct in the diet had linear effect on GGT (Pâ¯<â¯.001) and AST (Pâ¯<â¯.0001) enzymes as well as on total bilirubin (Pâ¯=â¯.05). In addition, the highest grape byproduct addition showed the highest consumption of hay (Pâ¯<â¯.01). Hay replacement by grape byproduct at up to 300â¯g/kg in the DM was satisfactory to weight gain and did not negatively affect feed intake and weight gain of growing lambs. Maintaining zinc:copper ratio in sheep diets is not effective in preventing liver damage caused by increased dietary copper concentrations over a period of 70â¯days.
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Cobre/intoxicação , Intoxicação por Metais Pesados/veterinária , Carneiro Doméstico/fisiologia , Vitis/química , Zinco/metabolismo , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Dieta/veterinária , Suplementos Nutricionais/análise , Feminino , Intoxicação por Metais Pesados/prevenção & controle , Zinco/administração & dosagemRESUMO
BACKGROUND: Individual-level measures of acculturation (e.g. age of immigration) have a complex relationship with psychiatric disorders. Fine-grained analyses that tap various acculturation dimensions and population subgroups are needed to generate hypotheses regarding the mechanisms of action for the association between acculturation and mental health. METHOD: Study participants were US Latinos (N = 6359) from Wave 2 of the 2004-2005 National Epidemiologic Survey of Alcohol and Related Conditions (N = 34 653). We used linear χ2 tests and logistic regression models to analyze the association between five acculturation dimensions and presence of 12-month DSM-IV mood/anxiety disorders across Latino subgroups (Mexican, Puerto Rican, Cuban, 'Other Latinos'). RESULTS: Acculturation dimensions associated linearly with past-year presence of mood/anxiety disorders among Mexicans were: (1) younger age of immigration (linear χ2 1 = 11.04, p < 0.001), (2) longer time in the United States (linear χ2 1 = 10.52, p < 0.01), (3) greater English-language orientation (linear χ2 1 = 14.57, p < 0.001), (4) lower Latino composition of social network (linear χ2 1 = 15.03, p < 0.001), and (5) lower Latino ethnic identification (linear χ2 1 = 7.29, p < 0.01). However, the associations were less consistent among Cubans and Other Latinos, and no associations with acculturation were found among Puerto Ricans. CONCLUSIONS: The relationship between different acculturation dimensions and 12-month mood/anxiety disorder varies across ethnic subgroups characterized by cultural and historical differences. The association between acculturation measures and disorder may depend on the extent to which they index protective or pathogenic adaptation pathways (e.g. loss of family support) across population subgroups preceding and/or following immigration. Future research should incorporate direct measures of maladaptive pathways and their relationship to various acculturation dimensions.
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Aculturação , Transtornos de Ansiedade/etnologia , Emigração e Imigração/estatística & dados numéricos , Hispânico ou Latino/estatística & dados numéricos , Transtornos do Humor/etnologia , Adolescente , Adulto , Fatores Etários , Idoso , Cuba/etnologia , Feminino , Inquéritos Epidemiológicos , Humanos , Masculino , Americanos Mexicanos/estatística & dados numéricos , Pessoa de Meia-Idade , Porto Rico/etnologia , Estados Unidos/etnologia , Adulto JovemRESUMO
Follicular non-Hodgkin's lymphoma (FL) is a nodal B lymphoid malignancy that originates from the germinal center of a lymph node. FL is the second most frequent lymphoma subtype. The course of the disease is usually characterised by a typically indolent clinical course, with a median survival rate of 8-10 years, although most patients relapse after treatment. Diagnosis should always be based on a surgical specimen like an excisional node lymph biopsy. The first-line treatment of FL will depend of extension disease, tumour burden, patient symptoms, performance status and also patient decision. The addition of rituximab to conventional chemotherapy has improved ORR, PFS and OS. As first line in patients that need treatment, a combination of chemotherapy with rituximab induction followed by 2 years of rituximab maintenance is the best option. High-dose chemotherapy with autologous stem-cell transplantation in first line has not shown improvement and is not recommended as first-line therapy. Before any treatment decision in relapsed patients, a repeat biopsy is mandatory to rule out a transformation into large cell aggressive lymphoma. Standard treatment is controversial, depends on the efficacy of prior treatment, duration of the time-to-relapse, patient's age and histological findings at relapse.
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Linfoma não Hodgkin/diagnóstico , Linfoma não Hodgkin/terapia , Guias de Prática Clínica como Assunto/normas , Ensaios Clínicos como Assunto , Terapia Combinada , Gerenciamento Clínico , Detecção Precoce de Câncer , Humanos , Oncologia , Estadiamento de Neoplasias , Prognóstico , Sociedades MédicasRESUMO
Hodgkin lymphoma (HL) is an uncommon B cell lymphoid malignancy representing approximately 10-15 % of all lymphomas. HL is composed of two distinct disease entities; the more commonly diagnosed classical HL and the rare nodular lymphocyte-predominant HL. An accurate assessment of the stage of disease and prognostic factors that identify patients at low or high risk for recurrence are used to optimize therapy. Patients with early stage disease are treated with combined modality strategies using abbreviated courses of combination chemotherapy followed by involved-field radiation therapy, while those with advanced stage disease receive a longer course of chemotherapy often without radiation therapy. High-dose chemotherapy (HDCT) followed by an autologous stem cell transplant (ASCT) is the standard of care for most patients who relapse following initial therapy. Brentuximab vedotin should be considered for patients who fail HDCT with ASCT.
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Doença de Hodgkin/diagnóstico , Doença de Hodgkin/terapia , Guias de Prática Clínica como Assunto/normas , Ensaios Clínicos como Assunto , Terapia Combinada , Gerenciamento Clínico , Detecção Precoce de Câncer , Humanos , Oncologia , Estadiamento de Neoplasias , Prognóstico , Sociedades MédicasRESUMO
A partir de estudios previos realizados sobre la anatomía del musculo bíceps femoral en el perro y con el objetivo de avanzar en la caracterización inmunohistoquímica y nerviosa de este musculo, estudiamos la existencia de subvolúmenes musculares. Utilizamos la disección roma en piezas musculares aisladas fijadas y tratadas con ácido nítrico al 25%. Las divisiones de las ramas del nervio isquiático involucradas fueron utilizadas como guía en la disección. Se determinaron tres subvolúmenes, uno involucrando a la cabeza caudal y dos en el vientre correspondiente a la cabeza craneal que fueron denominados caudal, proximal y distal respectivamente. Realizamos mediciones del largo de las fibras musculares, dirección, dirección y densidad macroscópica de las mismas para caracterizar los compartimientos. Estos subvolúmenes deberán tenerse en cuanta al realizar estudios electromiográficos del musculo bíceps femoral y al evaluar las alteraciones en la locomoción producto de las alteraciones nerviosas y musculares en el nervio pelviano del canino.
From previous studies on the anatomy of the femoral biceps muscle in the dog and with the aim of advancing in immunohistochemical and nervous characterization of this muscle, we studied the existence of subvolumes in muscle structure. We used blunt dissection in isolated muscle pieces fixed and treated with 25% nitric acid. The divisions of the sciatic nerve branches involved were used to guide the dissection. Three subvolumes were identified; one, involving the caudal head, and two in the cranial head belly; they were called caudal, proximal and distal respectively. We measured muscle fiber length, orientation, and macroscopic density to characterize these compartments. These subvolumes must be considered when muscle electromyography research is carried out, and for locomotion changes evaluation due to lesions in nerve and muscle structure of the canine pelvic limb.
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Animais , Cães/anatomia & histologia , Músculos Isquiossurais/anatomia & histologiaRESUMO
Although it is accepted that impaired placental perfusion in complicated pregnancy can slow fetal growth and programme an increased risk of cardiovascular dysfunction at adulthood, the relative contribution of reductions in fetal nutrition and in fetal oxygenation as the triggering stimulus remains unclear. By combining high altitude (HA) with the chick embryo model, we have previously isolated the direct effects of HA hypoxia on embryonic growth and cardiovascular development before hatching. This study isolated the effects of developmental hypoxia on cardiovascular function measured in vivo in conscious adult male and female chickens. Chick embryos were incubated, hatched and raised at sea level (SL, nine males and nine females) or incubated, hatched and raised at HA (seven males and seven females). At 6 months of age, vascular catheters were inserted under general anaesthesia. Five days later, basal blood gas status, basal cardiovascular function and cardiac baroreflex responses were investigated. HA chickens had significantly lower basal arterial PO2 and haemoglobin saturation, and significantly higher haematocrit than SL chickens, independent of the sex of the animal. HA chickens had significantly lower arterial blood pressure than SL chickens, independent of the sex of the animal. Although the gain of the arterial baroreflex was decreased in HA relative to SL male chickens, it was increased in HA relative to SL female chickens. We show that development at HA lowers basal arterial blood pressure and alters baroreflex sensitivity in a sex-dependent manner at adulthood.
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Doença da Altitude/fisiopatologia , Barorreflexo/fisiologia , Sistema Cardiovascular/fisiopatologia , Efeitos Tardios da Exposição Pré-Natal/etiologia , Doença da Altitude/complicações , Animais , Gasometria , Pressão Sanguínea , Bolívia , Embrião de Galinha , Feminino , Testes de Função Cardíaca , Hematócrito , Masculino , Gravidez , Fatores SexuaisRESUMO
introducción: la fisura labio palatina no sindrómica, NSCLP (del inglés Nonsyndromic cleft lip and palate) es una de las malformaciones congénitas más frecuentes tanto en Chile como en el resto del mundo. Presenta un modo de herencia multifactorial, en la que interactúan varios genes y el medio ambiente. Evidencias experimentales han demostrado la participación de Sonic hedgedhog (Shh) en la migración de las células de la cresta neural, en la transformación epitelio-mesénquima y en la formación de las estructurasmedias craneofaciales durante el desarrollo embrionario, es probable una asociación entre variantes de Shh y la NSCLP. Métodos: el objetivo de este trabajo fue evaluar las regiones exónicas e intrónicas adyacentes de Shh, en una muestra de 150 tríos caso-progenitores para hallar la asociación con NSCLP. Se utilizó el método PCR-RFLP para determinar la presencia de heterodúplex. Luego, se utilizó la técnica de Conformation Sensitive Gel Electrophoresis (CSGE) para ver la distorsión del ADN en los heterodúplex. Como método alternativo, se hizo un análisis de polimorfismos de un solo nucleótido (del inglés single-nucleotide polymorphism SNP) para determinar asociación entre NSCLP y Shh, para lo cual se utilizaron los SNP: rs1233555 y rs1233556, ubicados en el primer intrón de Shh. Resultados:no se observaron heterodúplex en ninguno de los segmentos de Shh analizados en 150 tríos, el análisis de SNP tampoco mostró asociación con Shh y FLPNS. conclusión: la no asociación puede deberse a que la frecuencia de distribución de los SNP en la población chilena es diferente a la de las poblaciones referidas, o a que el número de SNP analizados fue insuficiente, o la no inclusión para el análisis de otras regiones de Shh.
introduction: nonsyndromic cleft lip and palate (NSCLP) is one of the most common congenital malformations not only in Chile but also worldwide. It has a multifactorial inheritance pattern with interaction of several genes and the environment. Several experimental studies have proven the participation of Sonic hedgedhog (Shh) in the migration process of cells from the neural crest,in the epithelium-mesenchyme transformation, and in the formation of middle craniofacial structures during embryo development; an association between Shh variants and NSCLP is probable. Methods: the goal of this study was to evaluate both exonic and intronic regions adjacent to Shh, in a sample of 150 case-parent trios in order to find possible associations with NSCLP. The PCR-RFLP method was used to determine the presence of heteroduplex. Afterwards, the Conformation Sensitive Gel Electrophoresis (CSGE) technique wasused to visualize DNA distortion at the heteroduplexes. As an alternative method, a single-nucleotide polymorphism (SNP) analysis wasperformed in order to determine NSCLP-Shh associations, by means of these SNPs: rs1233555 and rs1233556, located at the first Shhintron. Results: no heteroduplexes were found in any of the analyzed Shh segments in 150 trios; SNP analysis did not show associations between Shh and NSCLP either. conclusions: this lack of association may be due to the fact that SNP distribution frequency among Chilean population is different to that of reference populations, or because the number of SNPs analyzed was not sufficient, or even because this study did not include other Shh regions.
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Humanos , Fenda Labial , Fissura Palatina , LábioRESUMO
El músculo bíceps femoral (BF) desempeña un rol fundamental en la biomecánica del miembro pelviano de los caninos, actuando en la flexoextensión de la articulación femorotibiorotuliana y la abducción del miembro. La evaluación correcta de su función es importante en la clínica veterinaria. Existen puntos controversiales entre los autores consultados acerca de la morfología de este músculo. El presente estudio tiene como objetivo discutir las distintas descripciones, reparando en las inserciones, la estructura e inervación del músculo. Se han realizado las disecciones bilaterales en 12 animales. La conservación de las piezas se realizó por inmersión en piletas, con una dilución de formol al 10 por ciento y ácido fenico al 4 por ciento en agua. Se utilizaron diferentes técnicas de abordaje al músculo en cuestión, para obtener distintas observaciones de las estructuras. Se halló que el músculo BF se origina por medio de dos cabezas, como queda implícito en su denominación, una cabeza craneal, más voluminosa y una cabeza caudal más pequeña. La primera originada en el ligamento sacrotuberal y en la superficie lateral de la tuberosidad isquiática. La cabeza caudal, se origina de la tuberosidad isquiática. Estas partes a pesar de hallarse estrechamente unidas se individualizan a nivel de los vientres musculares por medio de una delgada lámina de tejido conectivo que se extiende hasta el tercio distal del músculo. Respecto a la inervación, no se encontraron ramas del nervio glúteo caudal que inerven las partes al músculo.
The biceps femoris (BF) muscle plays a fundamental role in the biomechanics of the canine pelvic limb, acting in flexion extension on knee articulation and abduction of the limb. Correct evaluation plays an important role in veterinary clinic. There are controversial issues among the authors consulted about the morphology of this muscle. The present study aims to discuss the various descriptions proposed for this muscle, considering insertions, structure and innervation. Bilateral dissections of 12 animals were carried out. Pieces conservation was done by immersion in pools with 10 percent formaldehyde and 4 percent carbolic acid in water. Different techniques were used in order to achieve various observations of muscle structures. It was found that BF muscle originates by two heads, as is implicit in its name, a craneal head, bulkier and a smaller caudal head. The first takes origin in the sacrotuberal ligament and the lateral surface of the ischial tuberosity. The caudal head, originates from the ischial tuberosity. These parts are closely linked but they can be identified at the level of the muscle bellies through a thin layer of connective tissue that extends to the distal part of the muscle. Regarding the innervation, we found that no caudal gluteal nerve branches reach this muscle.
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Animais , Cães , Músculo Esquelético/anatomia & histologia , Coxa da Perna/anatomia & histologia , Cães/anatomia & histologia , Músculo Esquelético/inervação , Coxa da Perna/inervaçãoRESUMO
The partial contributions of reductions in fetal nutrition and oxygenation to slow fetal growth and a developmental origin of cardiovascular disease remain unclear. By combining high altitude with the chick embryo model, we have previously isolated the direct effects of high-altitude hypoxia on growth. This study isolated the direct effects of high-altitude hypoxia on cardiovascular development. Fertilized eggs from sea-level or high-altitude hens were incubated at sea level or high altitude. Fertilized eggs from sea-level hens were also incubated at high altitude with oxygen supplementation. High altitude promoted embryonic growth restriction, cardiomegaly and aortic wall thickening, effects which could be prevented by incubating eggs from high-altitude hens at sea level or by incubating eggs from sea-level hens at high altitude with oxygen supplementation. Embryos from high-altitude hens showed reduced effects of altitude incubation on growth restriction but not on cardiovascular remodeling. The data show that: (1) high-altitude hypoxia promotes embryonic cardiac and vascular disease already evident prior to hatching and that this is associated with growth restriction; (2) the effects can be prevented by increased oxygenation; and (3) the effects are different in embryos from sea-level or high-altitude hens.
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Amplicon vectors derived from herpes simplex virus type 1 were built to modify NMDA receptors by expressing antisense RNA for the essential NR1 subunit. Their ability to modify endogenous levels of NR1 was tested in cultures of rat embryo neocortical neurons. We studied behaviour and tested for expression in adult rats injected with those vectors into the dorsal hippocampus to find out which cells and how many appear involved in memory formation. Rats injected with vectors expressing NR1 antisense performed significantly worse than control rats in an inhibitory avoidance task. Immunohistochemistry was performed in brain slices from the same animals. The transduced cells represented 6-7% of pyramidal neurons in CA1, showing that a single gene knockdown of NR1 in a small number of neurons significantly impaired memory formation. Perhaps neurons undergoing synaptic plasticity are more susceptible to NR1 knockdown, and hence NMDAR are particularly required in those neurons undergoing synaptic plasticity during learning, or perhaps, and more likely, there is not a high level of redundancy in the hippocampal circuits involved, leading to the idea that a certain level of NR1 expression/availability appears necessary for memory formation in most of CA1 pyramidal neurons.
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Hipocampo/fisiologia , Aprendizagem/fisiologia , Neurônios/fisiologia , Receptores de N-Metil-D-Aspartato/fisiologia , Animais , Células Cultivadas , Expressão Gênica , Vetores Genéticos , Proteínas de Fluorescência Verde/genética , Herpesvirus Humano 1/genética , Hipocampo/citologia , Immunoblotting , Imuno-Histoquímica , Masculino , Memória/fisiologia , Microscopia Confocal , RNA Antissenso/genética , Ratos , Ratos Wistar , Receptores de N-Metil-D-Aspartato/genética , Proteínas Recombinantes de Fusão , TransfecçãoRESUMO
Background: Nonsyndromic cleft lip/palate (NSCLP) is a congenital malformation with the characteristics of a complex genetic trait. Based on experimental evidences as well as on association and linkage studies candidate genes TGFA, RARA and BCL3 have been postulated as being involved in the genetic etiology of this pathology. Aim: To test the possible association due to linkage disequilibrium between microsatellite markers located at less than 1cM from the three candidate genes and nonsyndromic cleft lip/palate using the case-parents trio design. Patients and Methods: The sample consisted of 58 case-parents trios. Two microsatellite markers, flanking each one of the candidate genes were analyzed by means of the polymerase chain reaction (PCR) with fluorescent labeled microsatellite markers. Electrophoresis of the PCR products was performed on a laser-fluorescent automatic DNA sequencer. Nonparametric ETDT was used to analyze the genotype data. Results: Significant linkage disequilibrium was detected between D2S443 (TGFA) and NSCLP. Significance was almost reached between D17S800 (RARA) and NSCLP. Alleles 239bp (D2S443) and 172bp (D17S800) showed significant preferential transmission from heterozygous parents to affected offspring. In the case of BCL3 both markers showed no significant results. Conclusions: The results of the present study do not show clear evidence that TGFA or RARA could be involved in the genetic etiology of NSCLP. Even though the importance of retinoic acid in the development of the embryo is well documented the results obtained for RARA are difficult to analyze. In relation to the possible role of BCL3 in NSCLP, recent information postulates that other genes located in the same chromosome region could be involved in NSCLP.
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Humanos , Fenda Labial/genética , Fissura Palatina/genética , Desequilíbrio de Ligação/genética , Repetições de Microssatélites/genética , Alelos , Chile , Marcadores Genéticos , Genótipo , Proteínas Proto-Oncogênicas/genética , Receptores do Ácido Retinoico/genética , Fatores de Transcrição , Fator de Crescimento Transformador alfa/genéticaRESUMO
Insecticide susceptibility in tobacco budworm, Heliothis virescens (F.) (Lepidoptera: Noctuidae), was determined for 8 yr (1991-2001) with larvae sampled from cotton in southern Tamaulipas, Mexico. Before 1996, when Bollgard cotton expressing the Cry1A(c) delta-endotoxin was introduced into the region, two important patterns were documented. The first was economically significant increases in resistance to certain insecticide groups. The second was occurrence of virtually complete control failures in the field during 1994 and 1995. The largest resistance changes were recorded for the type II pyrethroids cypermethrin and deltamethrin. These products are the most widely used products in the region. Resistance ratios for these products increased up to > 100-fold from 1991 to 1995. After 1996, the resistance levels declined. These findings did not occur with other products of scant use (e.g., permethrin, profenofos, and endosulfan) or low tobacco budworm efficacy coupled to a high use pattern (e.g., methyl parathion). This clear trend toward reversal of resistance to type II pyrethroids can be understood, in part, with respect to two factors: 1) the high adoption rate of transgenic cotton in the region, from 31.2% in the beginning (1996) to approximately 90% in 1998; this has considerably curbed the use of synthetic insecticides, with the attending loss of selection pressure on this pest; and 2) the potential immigration to the region of susceptible tobacco budworms from cultivated and wild suitable hosts as well as from transgenic cotton might have influenced the pest population as a whole. The influence of transgenic cotton on southern Tamaulipas can be more clearly seen by the drastic reduction of insecticide use to control this important pest. Now tobacco budworms in this region are susceptible to type II pyrethroids. Two effective and fundamentally different pest management tools are now available to cotton growers in southern Tamaulipas: transgenic cotton, coupled with careful use of pyrethroids, offers the possibility of sustainable and profitable cotton production.
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Gossypium/efeitos dos fármacos , Inseticidas/farmacologia , Mariposas/efeitos dos fármacos , Animais , Resistência a Inseticidas , México , Plantas Geneticamente Modificadas , Fatores de TempoRESUMO
Herpes simplex virus-derived amplicon vectors simultaneously expressing the open reading frame encoding NR1 subunit of the NMDA receptor, either in sense or antisense orientation, as well as the open reading frame encoding the green fluorescent protein (GFP), as distinct transcription units, were constructed. Vector expression in cells was demonstrated by GFP-fluorescence, immunofluorescence, Western blots and RT-PCR. The vectors were inoculated into the dorsal hippocampus of adult male rats, which were then trained for habituation to an open field and for inhibitory avoidance to a foot-shock. Those animals injected with vectors expressing NR1 protein showed habituation to a new environment, and achieved the criteria for a step-down inhibitory avoidance to a foot-shock. In contrast, animals injected with vectors carrying the NR1 open reading frame in antisense position, showed neither habituation nor appropriate performance in the inhibitory avoidance task. There was no evidence for motor impairment or motivational disturbance, since all the animals exhibit similar behavior and performance in the training sessions. Hence, the impaired performance might be due to either amnesia or disability to record events. Transgene expression in brain, as revealed by GFP fluorescence, was mainly observed in pyramidal cells of CA1, but also in CA3. Therefore, our results strongly support the participation of hippocampal NR1 subunit in habituation to a new environment, but also in recording events for the inhibitory avoidance task. Hence, amplicon vectors appear to be useful tools to modify endogenous gene expression at a defined period, in restricted brain regions, and should allow investigating in vivo functions of genes.
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Comportamento Animal/fisiologia , Técnicas de Transferência de Genes , Vetores Genéticos , Herpesvirus Humano 1/genética , Hipocampo/virologia , Oligonucleotídeos Antissenso/genética , Receptores de N-Metil-D-Aspartato/fisiologia , Animais , Linhagem Celular , Cricetinae , Expressão Gênica , Haplorrinos , Masculino , Aprendizagem em Labirinto/fisiologia , Plasmídeos , Ratos , Ratos Wistar , Receptores de N-Metil-D-Aspartato/genética , TransgenesRESUMO
1. The aim is to study some roles of the hippocampal NMDA receptor, by modifying the expression of the essential NR1 subunit, with temporal and spatial restrictions in the central nervous system (CNS) of the rat. 2. Due to their neurotropism and the size of inserts they can accomodate, herpes simplex virus type-1 (HSV-1) derived amplicon vectors were used to transfer sequences, either in sense (+) or antisense (-) orientations, of the NR1 subunit gene, or of the green fluorescent protein (GFP) gene, into the CNS. 3. Vector expression in cell lines was followed by GFP autofluorescence, immunofluorescence and western blot. 4. The vectors were inoculated into the dorsal hippocampus of adult male Wistar rats, which were evaluated for habituation to an open field, and then, for expression of the transgenes, by autofluorescence and western blot; the expression mainly happened in pyramidal cells of CA1. 5. The animals injected with vectors carrying the NR1(+) transgene showed habituation to the new environment, as also happened with rats injected with vectors carrying only the GFP transgene. 6. In contrast, animals injected with vectors carrying NR1(-) sequence, did not show habituation. This might be retrograde amnesia or disability to record the trace, suggesting that the NR1 subunit in the dorsal hippocampus, is involved in habituation to a new environment. 7. HSV-1 derived amplicon vectors appear to be useful tools to modify endogenous gene expression, at a defined period, in restricted regions of the CNS.
Assuntos
Habituação Psicofisiológica/genética , Hipocampo/metabolismo , Transtornos da Memória/genética , Terminações Pré-Sinápticas/metabolismo , Receptores de N-Metil-D-Aspartato/genética , Transmissão Sináptica/genética , Animais , Comportamento Animal/fisiologia , Cricetinae , Técnicas de Transferência de Genes , Vetores Genéticos/genética , Proteínas de Fluorescência Verde , Hipocampo/fisiopatologia , Imuno-Histoquímica , Aprendizagem/fisiologia , Proteínas Luminescentes , Masculino , Memória/fisiologia , Transtornos da Memória/metabolismo , Transtornos da Memória/fisiopatologia , Oligonucleotídeos Antissenso , Ratos , Ratos Wistar , Receptores de N-Metil-D-Aspartato/metabolismo , Simplexvirus/genéticaRESUMO
BACKGROUND: In the last 10 years type-I allergy against proteins from Hevea brasiliensis latex has become an acknowledged medical issue. Fruit-allergic patients represent one risk group for developing latex allergy. Class I chitinases have been identified from chestnut, avocado and banana as relevant allergens. The chitin binding (hevein) domain from these class I chitinases has been postulated to bear the important IgE binding epitopes. OBJECTIVE: To clone the cDNA of an allergenic latex class I chitinase, to express the recombinant protein and to determine its IgE cross-reactivity with hevein (Hev b 6.02). METHODS: A full-length cDNA coding for a class I chitinase has been isolated from Hevea latex RNA by reverse transcription followed by PCR. The chitinase encoding sequence has been subcloned into the pMAL expression vector and expressed in E. coli as a fusion protein to maltose binding protein. The highly enriched recombinant protein fraction has been tested for its IgE binding capacity in immunoblots and ELISA. Furthermore, the pathogenesis-related function of the recombinant protein was tested in a fungal growth inhibition assay. RESULTS: The Hevea brasiliensis latex chitinase, designated Hev b 11, displays 70% identity to the endochitinase from avocado and its hevein-domain 58% to hevein (Hev b 6.02). The recombinant Hev b 11-maltose binding protein is recognized by latex- and fruit-allergic patients with IgE binding in both, ELISA and immunoblots. Pre-incubation of sera with rHev b 11-maltose binding protein showed an overall 16% inhibition of subsequent binding to rHev b 6.02-maltose binding protein on solid phase. The growth of F. oxysporum was inhibited in a dose dependent manner by addition of rHev b 11-maltose binding protein to the culture. CONCLUSIONS: Hev b 11, a class I chitinase, is another allergen from Hevea latex with a chitin binding domain and displays a different IgE binding capacity compared with hevein.
Assuntos
Transportadores de Cassetes de Ligação de ATP , Alérgenos/química , Alérgenos/imunologia , Alérgenos/isolamento & purificação , Quitinases/química , Quitinases/imunologia , Quitinases/isolamento & purificação , Clonagem Molecular , Proteínas de Escherichia coli , Hevea/imunologia , Proteínas de Transporte de Monossacarídeos , Proteínas de Plantas/química , Sequência de Aminoácidos , Antígenos de Plantas , Sequência de Bases , Sítios de Ligação/imunologia , Proteínas de Transporte/imunologia , Proteínas de Transporte/isolamento & purificação , DNA Complementar/imunologia , Ensaio de Imunoadsorção Enzimática , Hipersensibilidade Alimentar/imunologia , Frutas/imunologia , Humanos , Immunoblotting , Imunoglobulina E/imunologia , Hipersensibilidade ao Látex/imunologia , Proteínas Ligantes de Maltose , Dados de Sequência Molecular , Proteínas de Plantas/imunologia , Proteínas de Plantas/isolamento & purificação , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/isolamento & purificaçãoRESUMO
We have analysed complete or partial VPI sequences of 31 foot-and-mouth disease (FMD) viruses belonging to serotypes A, O and C to determine the genetic relatedness of field strains of FMD virus (FMDV) that have circulated in Argentina between 1961 and 1994. Phylogenetic analysis, which also included 15 previously published Argentinean sequences and six reference strains, revealed that (i) FMD type A strains showed the highest genetic heterogeneity and could be divided into five lineages with a sequence divergence of 0.9-18.5% between strains (ii) most of the FMD type O viruses grouped in two clusters (within cluster sequence divergence ranging from 0.2% to 6.0%) circulating in Argentina since the early 1960s, and (iii) FMD type C viruses were grouped in two clusters with a 13.4% nucleotide sequence divergence between each cluster. The availability of sequence data for many more field isolates from the region will enable us to understand the genetic relationships between FMDV strains and to rapidly trace the source of an FMD outbreak for epidemiological surveillance.
Assuntos
Vírus da Febre Aftosa/classificação , Argentina/epidemiologia , Vírus da Febre Aftosa/genética , Epidemiologia Molecular , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Nucleotide sequence and phylogenetic analysis of the VP1 structural protein have been used extensively as diagnostic and epidemiological tools for foot and mouth disease virus (FMDV). In this report we have applied this methodology to the analysis of the VP1 coding sequence from FMDV strains isolated in Argentina during 1993-1994. The results demonstrated that the field isolates were related to the vaccine strains used at that time. However the involvement of the vaccine virus appeared to be different for outbreaks caused by FMD viruses type O or C. These data provide a database essential for determining the origin of new epizootics.