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INTRODUCTION: Uropathogenic Escherichia coli (UPEC) rely on the correlation of virulence expression with antimicrobial resistance to persist and cause severe urinary tract infections (UTIs). OBJECTIVES: We assessed the virulence pattern and prevalence among UPEC strains susceptible and resistant to multiple antimicrobial classes. METHODS: A total of 174 non-duplicate UPEC strains from patients with clinically significant UTIs were analysed for susceptibility to aminoglycoside, antifolate, cephalosporin, nitrofuran and quinolone antibiotics for the production of extended-spectrum ß-lactamases and for the presence of six virulence determinants encoding adhesins (afimbrial, Type 1 fimbriae, P and S-fimbriae) and toxins (cytotoxic necrotising factor and haemolysin). RESULTS: Relatively high resistance rates to nalidixic acid, ciprofloxacin, cephalothin and trimethoprim-sulfamethoxazole (82%, 78%, 62% and 59%, respectively) were observed. Fourteen distinct patterns were identified for the virulence determinants such as afaBC, cnfI, fimH, hylA, papEF and sfaDE. The toxin gene, cnfI (75.3%), was the second most prevalent marker to the adhesin, fimH (97.1%). The significant association of sfaDE/hylA (P < 0.01) among antimicrobial resistant and susceptible strains was also observed notwithstanding an overall greater occurrence of virulence factors among the latter. CONCLUSIONS: This study provides a snapshot of UPEC complexity in Jamaica and highlights the significant clonal heterogeneity among strains. Such outcomes emphasise the need for evidence-based strategies in the effective management and control of UTIs.
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Antibacterianos/farmacología , Farmacorresistencia Bacteriana , Infecciones Urinarias/microbiología , Escherichia coli Uropatógena/efectos de los fármacos , Escherichia coli Uropatógena/genética , Factores de Virulencia/genética , Humanos , Jamaica , Escherichia coli Uropatógena/aislamiento & purificación , beta-Lactamasas/análisisRESUMEN
We report on the growth and formation of single-layer boron nitride dome-shaped nanostructures mediated by small iron clusters located on flakes of hexagonal boron nitride. The nanostructures were synthesized in situ at high temperature inside a transmission electron microscope while the e-beam was blanked. The formation process, typically originating at defective step-edges on the boron nitride support, was investigated using a combination of transmission electron microscopy, electron energy loss spectroscopy and computational modelling. Computational modelling showed that the domes exhibit a nanotube-like structure with flat circular caps and that their stability was comparable to that of a single boron nitride layer.
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In this work, the cutting of carbon nanotubes is investigated using silver nanoparticles deposited on arc discharge multi-walled carbon nanotubes. The composite is subsequently heated in air to fabricate shortened multi-walled nanotubes. Complementary transmission electron microscopy and spectroscopy techniques shed light on the cutting mechanism. The nanotube cutting is catalysed by the fundamental mechanism based on the coordination of the silver atoms to the π-bonds of carbon nanotubes. As a result of the metal coordination, the strength of the carbon-carbon bond is reduced, promoting the oxidation of carbon at lower temperature when heated in air, or lowering the activation energy required for the removal of carbon atoms by electron beam irradiation, assuring in both cases the cutting of the nanotubes.
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BACKGROUND AND PURPOSE: Osseous pseudoprogression on MR imaging can mimic true progression in lesions treated with spine stereotactic radiosurgery. Our aim was to describe the prevalence and time course of osseous pseudoprogression to assist radiologists in the assessment of patients after spine stereotactic radiosurgery. MATERIALS AND METHODS: A secondary analysis of 2 prospective trials was performed. MRIs before and after spine stereotactic radiosurgery were assessed for response. "Osseous pseudoprogression" was defined as transient growth in signal abnormality centered at the lesion with a sustained decline on follow-up MR imaging that was not attributable to chemotherapy. RESULTS: From the initial set of 223 patients, 37 lesions in 36 patients met the inclusion criteria and were selected for secondary analysis. Five of the 37 lesions (14%) demonstrated osseous pseudoprogression, and 9 demonstrated progressive disease. There was a significant association between single-fraction therapy and the development of osseous pseudoprogression (P = .01), and there was a significant difference in osseous pseudoprogression-free survival between single- and multifraction regimens (P = .005). In lesions demonstrating osseous pseudoprogression, time-to-peak size occurred between 9.7 and 24.4 weeks after spine stereotactic radiosurgery (mean, 13.9 weeks; 95% CI, 8.6-19.1 weeks). The peak lesion size was between 4 and 10 mm larger than baseline. Most lesions returned to baseline size between 23 and 52.4 weeks following spine stereotactic radiosurgery. CONCLUSIONS: Progression on MR imaging performed between 3 and 6 months following spine stereotactic radiosurgery should be treated with caution because osseous pseudoprogression may be seen in more than one-third of these lesions. Single-fraction spine stereotactic radiosurgery may be associated with osseous pseudoprogression. The possibility of osseous pseudoprogression should be incorporated into the prospective criteria for assessment of local control following spine stereotactic radiosurgery.
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Radiocirugia , Columna Vertebral/patología , Columna Vertebral/cirugía , Anciano , Progresión de la Enfermedad , Femenino , Humanos , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Radiocirugia/métodosRESUMEN
OBJECTIVE: This study evaluated the ability of 0.8% neem leaf extract (NLE) to treat diabetes mellitus by assessing its effects on blood glucose, insulin levels and islet morphology in streptozotocin (STZ)-induced diabetic Sprague-Dawley rats. METHODS: Diabetes was induced in two to three-day old rat pups by STZ intraperitoneally (60 mg/kg), followed by a further 40 mg/kg dose 12-23 weeks later. The diabetic treated (DT) rats received 0.8% w/v NLE in tap water while diabetic control (DC) and normal control (NC) rats received water ad libitum. Body weight, water and chow consumption, and blood glucose were evaluated weekly. Blood and pancreas were collected at the end of the study to evaluate serum insulin and islet histology, respectively. RESULTS: Neem leaf extract (0.8%) improved weight gain and beta cell regeneration but did not reduce blood glucose. Serum insulin increased slightly in the treated group and three-fold in the DC group (p < 0.05). CONCLUSION: The results suggest that NLE has beta cell regenerating potential.
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BACKGROUND: A new policy requiring staff in clinical areas to vaccinate or wear a mask was implemented in British Columbia (BC) in the 2012/13 winter. This review assessed the impact of the policy on absenteeism in health care workers. METHODS: A retrospective cohort study of full-time HCW that worked prior to and during the 2012/13 influenza season in a health authority in BC. The rate of absenteeism due to all cause illness was compared between vaccinated and unvaccinated staff controlling for behaviors outside influenza season. RESULTS: Of the 10079 HCW, 77% were vaccinated. By comparison to absenteeism rates in the pre-influenza season, unvaccinated staff in winter had twice the increase in absenteeism due to all-cause illness than vaccinated staff. CONCLUSION: After controlling for baseline differences between those vaccinated and unvaccinated, influenza vaccination was associated with reduced absenteeism, saving the Health Authority substantial money. Having regular staff in attendance increases the quality of care.
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Absentismo , Personal de Salud , Programas de Inmunización , Gripe Humana/prevención & control , Vacunación/economía , Adulto , Colombia Británica , Estudios de Cohortes , Análisis Costo-Beneficio , Femenino , Personal de Salud/economía , Política de Salud/economía , Humanos , Programas de Inmunización/economía , Masculino , Máscaras , Estudios Retrospectivos , Estaciones del AñoRESUMEN
BACKGROUND: Transformation of indolent lymphomas (IL) to an aggressive histology (TIL) often results in a rapid clinical course, treatment refractoriness and shortened survival. Although rituximab-containing regimens (R-chemo) have become standard of care in CD20-positive TIL, the role of autologous stem-cell transplantation (ASCT) is still debated. The purpose of this study was to determine whether the outcome of TIL patients improved if they, at transformation, also received ASCT. Furthermore, we investigated the outcome of cases with histologically low- and high-grade components diagnosed either simultaneously or after a period of overt indolent disease. We also analyzed, whether prior rituximab treatment during the indolent course of the disease affected outcome after transformation. PATIENTS AND METHODS: Eighty-five patients (≤68 years) with histologically confirmed TIL were included. Five-year overall (OS) and progression-free survival (PFS) were calculated. Selected parameters were tested in a multivariate analysis. All analyses were conducted on three cohorts: (i) whole cohort (all TIL), (ii) patients with co-existing evidence of both indolent and aggressive histology at diagnosis (Composite/discordant TIL) and (iii) patients transformed after prolonged prior indolent disease (sequential TIL). RESULTS: Fifty-four patients (64%) received ASCT consolidation and 31 (36%) did not. Within the 'all TIL' cohort, the 5-year OS and PFS for R-chemo + ASCT versus R-chemo alone, were 67% versus 48% (P = 0.11) and 60% versus 30% (P = 0.02), respectively. Furthermore, in 'Composite/discordant TIL' R-chemo + ASCT showed no impact on OS (76% versus 67%; P = 0.66) or PFS (71% versus 62%; P = 0.54). Conversely, R-chemo + ASCT improved the outcome of 'sequential TIL' (OS 62% versus 36%; P = 0.07; PFS 53% versus 6%; P = 0.002), regardless of prior rituximab therapy. The beneficial effect of ASCT was significantly higher in patients who had not received rituximab at IL stage. CONCLUSIONS: ASCT improved the outcome in sequential, but not composite/discordant TIL. The beneficial impact of ASCT was greater in patients, who were rituximab-naïve at transformation.
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Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Terapia Combinada/métodos , Trasplante de Células Madre Hematopoyéticas/métodos , Linfoma/terapia , Adulto , Anciano , Transformación Celular Neoplásica/patología , Supervivencia sin Enfermedad , Femenino , Humanos , Linfoma/mortalidad , Linfoma/patología , Masculino , Persona de Mediana Edad , Rituximab/administración & dosificación , Trasplante AutólogoRESUMEN
In follicular lymphoma, nonmalignant immune cells are important. Follicular lymphoma depends on CD4+ cells, but CD8+ cells counteract it. We hypothesized that the presence of follicular lymphoma is associated with higher CD4+ than CD8+ cell numbers in the tumor microenvironment but not in the immune system. Using flow cytometry, pre-treatment and follow-up CD4/CD8 ratios were estimated in the bone marrow, blood and lymph nodes of untreated follicular lymphoma patients in two independent data sets (N(1)=121; N(2)=166). The ratios were analyzed for their relation with bone marrow lymphoma involvement. Bone marrows were also investigated with immunohistochemistry. In either data set, the bone marrow CD4/CD8 ratios were higher in bone marrows involved with lymphoma (P=0.043 and 0.0002, respectively). The mean CD4/CD8 ratio was 1.0 in uninvolved and 1.4 in involved bone marrows. Also higher in involved bone marrows were CD4/CD56 and CD3CD25/CD3 ratios. No blood or lymph node ratios differed between bone marrow-negative and -positive patients. Sequential samples showed increased bone marrow CD4/CD8 ratios in all cases of progression to bone marrow involvement. Immunohistochemistry showed CD4+, CD57+, programmed death-1+, forkhead box protein 3+ and CD21+ cells accumulated inside the lymphoma infiltrates, whereas CD8+, CD56+ and CD68+ cells were outside the infiltrates. This study provides evidence in vivo that the microenvironment changes upon follicular lymphoma involvement.
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BACKGROUND: Optimal treatment of young patients with high-risk diffuse large B-cell lymphoma (DLBCL) remains a matter of debate and requires improvement. The combination chemotherapy with cyclophosphamide, doxorubicin, vincristine and prednisone (CHOP) with addition of etoposide (CHOEP) has in other patient groups been shown to be effective. Further improvement has been accomplished with the use of rituximab in combination with the regimens every 2 weeks (R-CHOP-14, R-CHOEP-14). The aim of the present retrospective population-based study was to compare R-CHOP-14 with R-CHOEP-14 in a cohort of high-risk patients aged 18-60 years with two or more risk factors (stage III-IV, elevated lactate dehydrogenase levels, performance status 2-4). To our knowledge, this is the first study comparing these two regimens in this patient group. METHODS: We obtained data for the period 2004-2009 from the Danish Lymphoma Database. One hundred and fifty-nine patients were eligible to enter the study. Primary end point was overall survival (OS) and secondary end points were response to treatment, progression-free survival (PFS) and safety. RESULTS: Four-year OS was superior in the R-CHOEP-14 group: 75% compared with 62% for R-CHOP-14 (P=0.04). This superiority was also seen for PFS: 4-year PFS was 70% for the R-CHOEP-14 group compared with 58% for the R-CHOP-14 group (P=0.02). CONCLUSION: R-CHOEP-14 is a promising regimen for young patients with high-risk DLBCL with improved OS and PFS compared with R-CHOP-14.
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Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Linfoma de Células B Grandes Difuso/tratamiento farmacológico , Adulto , Anciano , Anticuerpos Monoclonales de Origen Murino/administración & dosificación , Ciclofosfamida/administración & dosificación , Dinamarca , Supervivencia sin Enfermedad , Doxorrubicina/administración & dosificación , Etopósido/administración & dosificación , Femenino , Humanos , Estimación de Kaplan-Meier , Linfoma de Células B Grandes Difuso/mortalidad , Masculino , Persona de Mediana Edad , Prednisona/administración & dosificación , Modelos de Riesgos Proporcionales , Sistema de Registros , Estudios Retrospectivos , Factores de Riesgo , Rituximab , Resultado del Tratamiento , Vincristina/administración & dosificaciónRESUMEN
OBJECTIVE: Pseudomonas aeruginosa produces multiple virulence factors that have been implicated in pathogenesis and quorum sensing. The aim of this study was to determine differences in the virulence factors of pigmented and non-pigmented P aeruginosa isolates. METHODS: Associations were assessed between pigment production (pyocyanin and pyoverdin) and production of DNase, elastase, lipase, protease, siderophore, twitching motility, antibiotic resistance patterns and virulence-associated genes in 57 non-duplicate P aeruginosa isolates from wounds, sputum, urine, high vaginal swab (HVS), ear, eye and respiratory tract swabs and aspirates of peritoneum and ulcers. RESULTS: Most (82.5%) of the isolates produced either pigment. Pigmented isolates produced more frequently and significant more (p < 0.05) DNase, elastase, lipase protease, and siderophore. Imipenem was the only antibiotic to which all isolates were susceptible (p < 0.05), while 93% and 32% were resistant to tetracycline and norfloxacin, respectively. There was however no significant difference between pigmented and non-pigmented isolates when antibiotic resistance was compared. While isolates had multiple virulence-associated genes, exoS (51%), rhlA (37%) and rhlB (46%) were the predominant genes detected. Except for exoY, genes were present in pigmented isolates more frequently than in non-pigmented isolates. CONCLUSION: The results of this study suggest that antibiotic resistance per se might not be associated with the pigment production in P aeruginosa. However pigment production appeared to be more significantly associated with multi-drug resistance, presence of virulence-associated genes, and expression of certain virulence factors, most notably elastase, protease, siderophore and DNase activity. Since pigment production is easy to determine, this might to be a good starting point to identify the virulence status of an isolate.
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Antibacterianos/farmacología , Norfloxacino/farmacología , Pseudomonas aeruginosa/metabolismo , Pseudomonas aeruginosa/patogenicidad , Tetraciclina/farmacología , ADP Ribosa Transferasas/genética , Proteínas Bacterianas/genética , Toxinas Bacterianas/genética , Distribución de Chi-Cuadrado , Farmacorresistencia Bacteriana , Regulación Bacteriana de la Expresión Génica , Humanos , Pruebas de Sensibilidad Microbiana , Oligopéptidos/metabolismo , Fenotipo , Reacción en Cadena de la Polimerasa , Pseudomonas aeruginosa/aislamiento & purificación , Piocianina/metabolismoRESUMEN
Free fatty acids (FFAs) exert complex actions on pancreatic ß-cells. Typically, an initial potentiation of insulin release is followed by a gradual impairment of ß-cell function, the latter effect being of possible relevance to hyperlipidaemia in type 2 diabetes mellitus. The molecular actions of FFAs are poorly understood. The present study investigated the acute effects of saturated FFAs on electrophysiological responses of rat pancreatic ß-cells. Membrane potential and KATP channel activity were recorded using the perforated patch technique. Volume-regulated anion channel (VRAC) activity was assessed from conventional whole-cell recordings. Cell volume regulation was measured using a video-imaging technique. Addition of octanoate caused a transient potentiation of glucose-induced electrical activity, followed by a gradual hyper-polarisation and a prolonged inhibition of electrical activity. Octanoate caused an initial increase in VRAC activity followed by a secondary inhibition coinciding with increased KATP channel activity. Similar effects were observed with palmitate and 2-bromopalmitate whereas butyrate was virtually ineffective. Octanoate and palmitate also exerted a dual effect on electrical activity evoked by tolbutamide. Octanoate significantly attenuated cell volume regulation in hypotonic solutions, consistent with VRAC inhibition. It is concluded that medium and long chain FFAs have a dual action on glucose-induced electrical activity in rat pancreatic ß-cells: an initial stimulatory effect followed by a secondary inhibition. These effects appear to be the result of reciprocal actions on VRAC and KATP channel currents, and could contribute towards the stimulatory and inhibitory actions of FFAs on pancreatic ß-cell function.
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Potenciales de Acción/fisiología , Ácidos Grasos/farmacología , Ácidos Grasos/fisiología , Células Secretoras de Insulina/fisiología , Canales KATP/fisiología , Canales Aniónicos Dependientes del Voltaje/fisiología , Potenciales de Acción/efectos de los fármacos , Animales , Células Cultivadas , Fenómenos Electromagnéticos , Femenino , Células HEK293 , Humanos , Células Secretoras de Insulina/efectos de los fármacos , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
BACKGROUND: Leptospirosis, a spirochetal zoonosis, is considered an occupational disease of persons engaged in agriculture, sewage works, forestry, and butchery. OBJECTIVE: To determine the environmental sources and the knowledge, attitude and practices for leptospirosis among butchers and slaughterhouse workers, as well as the seroprevalence of leptospirosis among cattle and pigs presented for slaughter. METHODS: Using an interviewer administered questionnaire, all 110 butchers and other slaughterhouse workers in the parishes of Kingston and St. Andrew, Jamaica were surveyed. In addition, 179 blood samples from animals presented for slaughter were tested for anti-Leptospira antibodies using the microscopic agglutination test (MAT). RESULTS: Analyses indicated that people with the studied occupations are at risk for developing leptospirosis due to several environmental risk factors that exist in slaughterhouses. Among the risk factors, limited knowledge of the disease and its transmission, lower educational level attained, younger age and unhealthy behaviors (e.g., hand washing and improper or lack of use of personal protective gears), presence of stray dogs and rodents, and inadequate maintenance of physical plants, were found to be important. Of the total number of animal samples tested, 20 (11%) were positive. Canicola and Hardjo (among cattle) and Bratislava (among pigs) were the major seroreactors. CONCLUSION: Butchers and slaughterhouse workers engaged in animal handling and slaughtering could be frequently exposed to leptospirosis, and hence control strategies targeting at these populations should be implemented.
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Conocimientos, Actitudes y Práctica en Salud , Leptospirosis/prevención & control , Enfermedades Profesionales/prevención & control , Salud Laboral/normas , Mataderos , Adolescente , Adulto , Factores de Edad , Anciano , Anciano de 80 o más Años , Animales , Anticuerpos Antibacterianos/sangre , Bovinos , Enfermedades de los Bovinos/epidemiología , Escolaridad , Femenino , Desinfección de las Manos , Humanos , Jamaica/epidemiología , Leptospira/inmunología , Leptospirosis/epidemiología , Leptospirosis/veterinaria , Masculino , Persona de Mediana Edad , Enfermedades Profesionales/epidemiología , Ropa de Protección , Factores de Riesgo , Estudios Seroepidemiológicos , Encuestas y Cuestionarios , Porcinos , Enfermedades de los Porcinos/epidemiología , Adulto JovenRESUMEN
OBJECTIVE: Quinolone resistance is usually caused by various chromosomal mutations, but has been more recently associated with plasmids which carry the qnr determinant. The aim of this study is to investigate the prevalence of qnr genes in clinical isolates of Enterobacteriaceae in Jamaica. METHODS: A total of 255 non-duplicate fluoroquinolone-resistant Enterobacteriaceae clinical isolates, comprising 232 Escherichia coli, 20 Klebsiella species and three Enterobacter spp were collected between October 2007 and November 2008 from hospitalized patients in Jamaica. The presence of the qnr gene was screened by PCR using specific primers for qnrA, qnrB and qnrS in extracted plasmid DNA. RESULTS: Eighty-three (32.5%) of these isolates were qnr-positive, of which 47.0% housed the qnrA gene only, 1.2% qnrB and 9.6% qnrS only. Another 36.1% possessed both qnrA and qnrS genes. Approximately 30% of the quinolone-resistant E coli isolates harboured the qnr gene while 50% Klebsiella spp and all Enterobacter spp were positive. CONCLUSION: The emergence of qnr-mediated quinolone resistance among clinical Enterobacteriaceae isolates is described for the first time in Jamaica.
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Proteínas Bacterianas/genética , Enterobacteriaceae/efectos de los fármacos , Fluoroquinolonas/farmacología , Farmacorresistencia Bacteriana/genética , Enterobacteriaceae/genética , Escherichia coli/genética , Humanos , Jamaica , Klebsiella/genética , Plásmidos/genéticaRESUMEN
Ultramicrotomy, focused ion beam scanning electron microscopy (FIBSEM) and cryogenic FIBSEM (cryo-FIBSEM) techniques, as developed for the controlled cross-sectioning of mesenchymal stem cells (MSCs) and human osteoblasts (HObs) on titanium (Ti) substrates for transmission electron microscopy (TEM) investigation, are compared. Conventional ultramicrotomy has been used to section cells on Ti-foil substrates embedded in resin, but significant problems with cell detachment using this technique restricted its general applicability. Conventional FIBSEM 'lift-out' procedures were found to be effective for the preparation of uniform sections of fixed and dehydrated cell/Ti specimens, but the control of cell staining remains an issue. Cryo-FIBSEM procedures used with an 'H-bar' sample geometry enabled the sectioning of fixed and hydrated cell/Ti specimens, but issues remain over ion beam-induced artefacts and control of frost on the sample foils.
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Microscopía Electrónica de Transmisión/métodos , Microtomía/métodos , Manejo de Especímenes/métodos , Microscopía por Crioelectrón/métodos , Humanos , Microscopía Electrónica de Rastreo/métodos , Osteoblastos/ultraestructura , Células Madre/ultraestructura , TitanioRESUMEN
We report experimental and theoretical studies of magnetic domain walls in an in-plane magnetized (Ga,Mn)As dilute moment ferromagnetic semiconductor. Our high-resolution electron holography technique provides direct images of domain wall magnetization profiles. The experiments are interpreted based on microscopic calculations of the micromagnetic parameters and Landau-Lifshitz-Gilbert simulations. We find that the competition of uniaxial and biaxial magnetocrystalline anisotropies in the film is directly reflected in orientation dependent wall widths, ranging from approximately 40 to 120 nm. The domain walls are of the Néel type and evolve from near-90 degrees walls at low temperatures to large angle [11[over ]0]-oriented walls and small angle [110]-oriented walls at higher temperatures.
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Development of high-yielding wheat varieties with good end-use quality has always been a major concern for wheat breeders. To genetically dissect quantitative trait loci (QTLs) for yield-related traits such as grain yield, plant height, maturity, lodging, test weight and thousand-grain weight, and for quality traits such as grain and flour protein content, gluten strength as evaluated by mixograph and SDS sedimentation volume, an F1-derived doubled haploid (DH) population of 185 individuals was developed from a cross between a Canadian wheat variety "AC Karma" and a breeding line 87E03-S2B1. A genetic map was constructed based on 167 marker loci, consisting of 160 microsatellite loci, three HMW glutenin subunit loci: Glu-A1, Glu-B1 and Glu-D1, and four STS-PCR markers. Data for investigated traits were collected from three to four environments in Manitoba, Canada. QTL analyses were performed using composite interval mapping. A total of 50 QTLs were detected, 24 for agronomic traits and 26 for quality-related traits. Many QTLs for correlated traits were mapped in the same genomic regions forming QTL clusters. The largest QTL clusters, consisting of up to nine QTLs, were found on chromosomes 1D and 4D. HMW glutenin subunits at Glu-1 loci had the largest effect on breadmaking quality; however, other genomic regions also contributed genetically to breadmaking quality. QTLs detected in the present study are compared with other QTL analyses in wheat.
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Productos Agrícolas/genética , Haploidia , Sitios de Carácter Cuantitativo , Triticum/genética , Análisis de Varianza , Canadá , Mapeo Cromosómico , Productos Agrícolas/anatomía & histología , Productos Agrícolas/crecimiento & desarrollo , Cruzamientos Genéticos , Ambiente , Marcadores Genéticos , Genotipo , Escala de Lod , Triticum/anatomía & histología , Triticum/crecimiento & desarrolloRESUMEN
Potassium chloride ion cotransporters (KCCs) are part of a family of transporters classically described as being involved in cell volume regulation. Recently, KCC2 has been shown to have a role in the development of the inhibitory actions of amine transmitters, whereas KCC3 also plays a fundamental role in the development and function of the central and peripheral nervous system. We have re-assessed the expression of each of the known KCCs in the rat forebrain using RT-PCR and in situ hybridisation histochemistry. As well as confirming the widespread expression of KCC1 and KCC2 throughout the brain, we now show a more restricted expression of KCC3a in the hippocampus, choroid plexus and piriform cortex, as well as KCC4 in the choroid plexus and the suprachiasmatic nucleus of the hypothalamus. The expression of KCC4 in the latter and KCC2 in the lateral hypothalamic and ventromedial hypothalamic nuclei suggests that these cotransporters may have selective roles in neuroendocrine or homeostatic functions. Finally, we demonstrate the existence of a truncated splice variation of KCC3a in the rat that appears to be expressed exclusively in neurons (as is KCC2), whereas the native form of KCC3a and KCC4 appears to be expressed in glial cells.
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Expresión Génica/fisiología , Prosencéfalo/metabolismo , Simportadores/metabolismo , Animales , Animales Recién Nacidos , Northern Blotting/métodos , Células Cultivadas , Técnicas de Cocultivo/métodos , Embrión de Mamíferos , Inmunohistoquímica/métodos , Hibridación in Situ/métodos , Masculino , Ratones , Ratones Endogámicos C57BL , Neuroglía/química , Neuroglía/metabolismo , Neuronas/metabolismo , Fosfopiruvato Hidratasa/metabolismo , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Simportadores/genéticaRESUMEN
The aim of this paper is to develop a broadly-applicable and self-consistent thin-film biphasic modelling framework for the full crawling cycle of a single animal cell. A hierarchy of thin-film two-phase 'reactive flow' models is derived; between them these cover a wide range of biologically relevant parameter regimes. The mathematical properties and biological implications of the resulting systems of high-order nonlinear degenerate parabolic-elliptic evolution equations are investigated. Linear-stability arguments suggest the formation of highly localized regions of high or low network density associated with small irregular oscillations or 'ruffling' of the plasma membrane. Local analyses at the contact line identify the classes of admissible contact-line conditions, through which we study for the first time the effect on the cell-scale motion of the 'mesoscopic' contact-line physics, which consists of the chemical and mechanical mechanisms for protrusive and retractive force generation near the outer cell periphery. One of the formulations is used to develop a minimal model for cell body translocation over a thin pseudopod, which predicts that myosin-driven contraction is not essential for rapid translocation. An analytic prediction for the translocation speed is given in terms of the network viscosity and slip coefficient (a parameter measuring the adhesion strength), of the membrane tension and of the thicknesses of the pseudopod and actin cortex; this is in good agreement with the translocation speed of osteoblasts on biomaterial substrates commonly used for orthopaedic implants. Limitations of the modelling approach and directions for future work are outlined.
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Movimiento Celular/fisiología , Modelos Biológicos , Actinas/fisiología , Animales , Fenómenos Biomecánicos , Adhesión Celular/fisiología , Citoesqueleto/fisiología , Humanos , Osteoblastos/fisiología , Seudópodos/fisiologíaRESUMEN
The epithelial cells of the choroid plexuses secrete cerebrospinal fluid (CSF), by a process which involves the transport of Na(+), Cl(-) and HCO(3)(-) from the blood to the ventricles of the brain. The unidirectional transport of ions is achieved due to the polarity of the epithelium, i.e. the ion transport proteins in the blood-facing (basolateral) membrane are different to those in the ventricular (apical) membrane. The movement of ions creates an osmotic gradient which drives the secretion of H(2)O. A variety of methods (e.g. isotope flux studies, electrophysiological, RT-PCR, in situ hybridization and immunocytochemistry) have been used to determine the expression of ion transporters and channels in the choroid plexus epithelium. Most of these transporters have now been localized to specific membranes. For example, Na(+)-K(+)ATPase, K(+) channels and Na(+)-2Cl(-)-K(+) cotransporters are expressed in the apical membrane. By contrast the basolateral membrane contains Cl(-)- HCO(3) exchangers, a variety of Na(+) coupled HCO(3)(-) transporters and K(+)-Cl(-) cotransporters. Aquaporin 1 mediates water transport at the apical membrane, but the route across the basolateral membrane is unknown. A model of CSF secretion by the mammalian choroid plexus is proposed which accommodates these proteins. The model also explains the mechanisms by which K(+) is transported from the CSF to the blood.
Asunto(s)
Sistema Nervioso Central/fisiología , Líquido Cefalorraquídeo/metabolismo , Plexo Coroideo/fisiología , Células Epiteliales/fisiología , Equilibrio Hidroelectrolítico/fisiología , Animales , Membrana Celular/fisiología , Sistema Nervioso Central/anatomía & histología , Plexo Coroideo/metabolismo , Células Epiteliales/metabolismo , Humanos , Iones/metabolismo , Proteínas de Transporte de Membrana/fisiología , Modelos BiológicosRESUMEN
AIMS/HYPOTHESIS: Hypoglycaemic sulphonylureas are thought to stimulate insulin release by binding to a sulphonylurea receptor, closing K(ATP) channels and inducing electrical activity. However, the fact that these drugs stimulate insulin release at high glucose concentrations where K(ATP) channels are closed suggests additional ionic actions. The aim of this study was to test the hypothesis that sulphonylureas influence the current of the glucose- and volume-regulated anion channel. METHODS: Electrical and ion-channel activity were recorded in isolated rat beta cells using the patch-clamp technique. (86)Rb(+) efflux was measured using intact islets. Beta cell volume was measured using a video-imaging technique. RESULTS: In the absence of glucose, tolbutamide (100 micromol/l) transiently depolarised the cells. In the presence of glucose (5 mmol/l), tolbutamide evoked a sustained period of electrical activity, whilst at 10 mmol/l glucose, the drug evoked a pronounced 'silent' depolarisation. In the absence of glucose, tolbutamide inhibited (86)Rb(+) efflux. However, at 10 mmol/l glucose, tolbutamide induced a transient stimulation of efflux. Tolbutamide potentiated the whole-cell volume-regulated anion conductance in a glucose-dependent manner with an EC(50) of 85 micromol/l. In single channel recordings, tolbutamide increased the channel-open probability. Tolbutamide caused beta cell swelling in the presence of glucose, but not in its absence. CONCLUSIONS/INTERPRETATION: Tolbutamide can induce beta cell electrical activity by potentiating the glucose- and volume-regulated anion channel current. This effect is probably not due to a direct effect of the drug on the channel, but could be secondary to a metabolic action in the beta cell.