RESUMEN
BACKGROUND: The purpose of this study was to compare the outcomes of infants with giant omphalocele (GO) born in two different epochs over two decades at a single institution. Specifically, it examined whether the utilization of selective pulmonary vasodilators and extracorporeal membrane oxygenator (ECMO) in the management of pulmonary hypertension in the second epoch were associated with improved outcomes. METHODS: The medical records of all patients diagnosed with GO at a large children's hospital from January 1, 1996 to December 31, 2016 were reviewed and divided into two epochs. Patients were classified as having an isolated GO or GO with minor or major associated anomalies. GO was defined as a defect more than or equal to 5âcm in size and/or liver in the sac. RESULTS: During the study period, 59 infants with GO were identified. The duration of invasive mechanical ventilation was significantly shorter among the survivors from the second epoch (pâ=â0.03), with none greater than seven days. There were no significant differences in the outcomes of survival to NICU discharge and length of stay (LOS) between infants in the two epochs. CONCLUSIONS: Infants with GO who required invasive mechanical ventilation for more than seven days did not survive in the second epoch. Survival did not improve with use of selective pulmonary vasodilators and ECMO. This information could be shared with families during prenatal and postnatal counselling to facilitate informed decision making regarding goals of care.
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Hernia Umbilical , Embarazo , Femenino , Niño , Humanos , Lactante , Hernia Umbilical/terapia , Hernia Umbilical/complicaciones , Hernia Umbilical/diagnóstico , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
Current treatments for xerostomia/dry mouth are palliative and largely ineffective. A permanent clinical resolution is being developed to correct hyposalivation using implanted hydrogel-encapsulated salivary human stem/progenitor cells (hS/PCs) to restore functional salivary components and increase salivary flow. Pluripotent epithelial cell populations derived from hS/PCs, representing a basal stem cell population in tissue, can differentiate along either secretory acinar or fluid-transporting ductal lineages. To develop tissue-engineered salivary gland replacement tissues, it is critical to reliably identify cells in tissue and as they enter these alternative lineages. The secreted protein α-amylase, the transcription factor MIST1, and aquaporin-5 are typical markers for acinar cells, and K19 is the classical ductal marker in salivary tissue. We found that early ductal progenitors derived from hS/PCs do not express K19, and thus earlier markers were needed to distinguish these cells from acinar progenitors. Salivary ductal cells express distinct polarity complex proteins that we hypothesized could serve as lineage biomarkers to distinguish ductal cells from acinar cells in differentiating hS/PC populations. Based on our studies of primary salivary tissue, both parotid and submandibular glands, and differentiating hS/PCs, we conclude that the apical marker MUC1 along with the polarity markers INADL/PATJ and SCRIB reliably can identify ductal cells in salivary glands and in ductal progenitor populations of hS/PCs being used for salivary tissue engineering. Other markers of epithelial maturation, including E-cadherin, ZO-1, and partition complex component PAR3, are present in both ductal and acinar cells, where they can serve as general markers of differentiation but not lineage markers.
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Proteínas de la Membrana , Mucina-1 , Glándulas Salivales , Proteínas Supresoras de Tumor , Xerostomía , Células Acinares/metabolismo , Biomarcadores/metabolismo , Diferenciación Celular , Células Cultivadas , Células Epiteliales , Humanos , Proteínas de la Membrana/metabolismo , Mucina-1/metabolismo , Glándulas Salivales/metabolismo , Proteínas de Uniones Estrechas/metabolismo , Proteínas Supresoras de Tumor/metabolismo , Xerostomía/terapiaRESUMEN
Decoding the genetic basis of lignocellulosic inhibitor tolerance in Saccharomyces cerevisiae is crucial for rational engineering of bioethanol strains with enhanced robustness. The genetic diversity of natural strains present an invaluable resource for the exploration of complex traits of industrial importance from a pan-genomic perspective to complement the limited range of specialised, tolerant industrial strains. Natural S. cerevisiae isolates have lately garnered interest as a promising toolbox for engineering novel, genetically encoded tolerance phenotypes into commercial strains. To this end, we investigated the genetic basis for lignocellulosic inhibitor tolerance of natural S. cerevisiae isolates. A total of 12 quantitative trait loci underpinning tolerance were identified by next-generation sequencing linked bulk-segregant analysis of superior interbred pools. Our findings corroborate the current perspective of lignocellulosic inhibitor tolerance as a multigenic, complex trait. Apart from a core set of genetic variants required for inhibitor tolerance, an additional genetic background-specific response was observed. Functional analyses of the identified genetic loci revealed the uncharacterised ORF, YGL176C and the bud-site selection XRN1/BUD13 as potentially beneficial alleles contributing to tolerance to a complex lignocellulosic inhibitor mixture. We present evidence for the consideration of both regulatory and coding sequence variants for strain improvement.
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Lignina/antagonistas & inhibidores , Sitios de Carácter Cuantitativo , Proteínas de Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/genética , Alelos , Ingeniería Genética , Variación Genética , Secuenciación de Nucleótidos de Alto Rendimiento , Herencia Multifactorial , FenotipoRESUMEN
Cell transplantation of autologous adult biopsies, grown ex vivo as epithelial organoids or expanded as spheroids, are proposed treatments to regenerate damaged branching organs. However, it is not clear whether transplantation of adult organoids or spheroids alone is sufficient to initiate a fetal-like program of branching morphogenesis in which coordinated branching of multiple cell types including nerves, mesenchyme and blood vessels occurs. Yet this is an essential concept for the regeneration of branching organs such as lung, pancreas, and lacrimal and salivary glands. Here, we used factors identified from fetal organogenesis to maintain and expand adult murine and human epithelial salivary gland progenitors in non-adherent spheroid cultures, called salispheres. These factors stimulated critical developmental pathways, and increased expression of epithelial progenitor markers such as Keratin5, Keratin14, FGFR2b and KIT. Moreover, physical recombination of adult salispheres in a laminin-111 extracellular matrix with fetal salivary mesenchyme, containing endothelial and neuronal cells, only induced branching morphogenesis when neurturin, a neurotrophic factor, was added to the matrix. Neurturin was essential to improve neuronal survival, axon outgrowth, innervation of the salispheres, and resulted in the formation of branching structures with a proximal-distal axis that mimicked fetal branching morphogenesis, thus recapitulating organogenesis. Epithelial progenitors were also maintained, and developmental differentiation programs were initiated, showing that the fetal microenvironment provides a template for adult epithelial progenitors to initiate branching and differentiation. Further delineation of secreted and physical cues from the fetal niche will be useful to develop novel regenerative therapies that instruct adult salispheres to resume a developmental-like program in vitro and to regenerate branching organs in vivo.
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Epitelio/inervación , Laminina/metabolismo , Neurturina/metabolismo , Glándulas Salivales/citología , Esferoides Celulares/citología , Células Madre/citología , Adulto , Animales , Materiales Biocompatibles/metabolismo , Células Cultivadas , Células Epiteliales/citología , Células Epiteliales/metabolismo , Epitelio/crecimiento & desarrollo , Epitelio/metabolismo , Femenino , Humanos , Ratones Endogámicos ICR , Neurogénesis , Glándulas Salivales/crecimiento & desarrollo , Glándulas Salivales/metabolismo , Esferoides Celulares/metabolismo , Células Madre/metabolismo , Ingeniería de TejidosRESUMEN
Strains of Saccharomyces cerevisiae with improved tolerance to plant hydrolysates are of utmost importance for the cost-competitive production of value-added chemicals and fuels. However, engineering strategies are constrained by a lack of understanding of the yeast response to complex inhibitor mixtures. Natural S. cerevisiae isolates display niche-specific phenotypic and metabolic diversity, encoded in their DNA, which has evolved to overcome external stresses, utilise available resources and ultimately thrive in their challenging environments. Industrial and laboratory strains, however, lack these adaptations due to domestication. Natural strains can serve as a valuable resource to mitigate engineering constraints by studying the molecular mechanisms involved in phenotypic variance and instruct future industrial strain improvement to lignocellulosic hydrolysates. We, therefore, investigated the proteomic changes between two natural S. cerevisiae isolates when exposed to a lignocellulosic inhibitor mixture. Comparative shotgun proteomics revealed that isolates respond by regulating a similar core set of proteins in response to inhibitor stress. Furthermore, superior tolerance was linked to NAD(P)/H and energy homeostasis, concurrent with inhibitor and reactive oxygen species detoxification processes. We present several candidate proteins within the redox homeostasis and energy management cellular processes as possible targets for future modification and study. Data are available via ProteomeXchange with identifier PXD010868.
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Lignina/toxicidad , Proteoma/análisis , Proteínas de Saccharomyces cerevisiae/análisis , Saccharomyces cerevisiae/química , Saccharomyces cerevisiae/efectos de los fármacos , Estrés Fisiológico , Tolerancia a Medicamentos , Proteómica , Saccharomyces cerevisiae/aislamiento & purificación , Saccharomyces cerevisiae/fisiologíaRESUMEN
BACKGROUND: Volumetric muscle loss caused by trauma or after tumour surgery exceeds the natural regeneration capacity of skeletal muscle. Hence, the future goal of tissue engineering (TE) is the replacement and repair of lost muscle tissue by newly generating skeletal muscle combining different cell sources, such as myoblasts and mesenchymal stem cells (MSCs), within a three-dimensional matrix. Latest research showed that seeding skeletal muscle cells on aligned constructs enhance the formation of myotubes as well as cell alignment and may provide a further step towards the clinical application of engineered skeletal muscle. In this study the myogenic differentiation potential of MSCs upon co-cultivation with myoblasts and under stimulation with hepatocyte growth factor (HGF) and insulin-like growth factor-1 (IGF-1) was evaluated. We further analysed the behaviour of MSC-myoblast co-cultures in different 3D matrices. RESULTS: Primary rat myoblasts and rat MSCs were mono- and co-cultivated for 2, 7 or 14 days. The effect of different concentrations of HGF and IGF-1 alone, as well as in combination, on myogenic differentiation was analysed using microscopy, multicolour flow cytometry and real-time PCR. Furthermore, the influence of different three-dimensional culture models, such as fibrin, fibrin-collagen-I gels and parallel aligned electrospun poly-ε-caprolacton collagen-I nanofibers, on myogenic differentiation was analysed. MSCs could be successfully differentiated into the myogenic lineage both in mono- and in co-cultures independent of HGF and IGF-1 stimulation by expressing desmin, myocyte enhancer factor 2, myosin heavy chain 2 and alpha-sarcomeric actinin. An increased expression of different myogenic key markers could be observed under HGF and IGF-1 stimulation. Even though, stimulation with HGF/IGF-1 does not seem essential for sufficient myogenic differentiation. Three-dimensional cultivation in fibrin-collagen-I gels induced higher levels of myogenic differentiation compared with two-dimensional experiments. Cultivation on poly-ε-caprolacton-collagen-I nanofibers induced parallel alignment of cells and positive expression of desmin. CONCLUSIONS: In this study, we were able to myogenically differentiate MSC upon mono- and co-cultivation with myoblasts. The addition of HGF/IGF-1 might not be essential for achieving successful myogenic differentiation. Furthermore, with the development of a biocompatible nanofiber scaffold we established the basis for further experiments aiming at the generation of functional muscle tissue.
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Diferenciación Celular/efectos de los fármacos , Factor de Crecimiento de Hepatocito/farmacología , Factor I del Crecimiento Similar a la Insulina/farmacología , Células Madre Mesenquimatosas/citología , Músculo Esquelético/fisiología , Mioblastos/citología , Ingeniería de Tejidos/métodos , Animales , Biomarcadores/metabolismo , Células Cultivadas , Técnicas de Cocultivo , Colágeno Tipo I/farmacología , Citometría de Flujo , Masculino , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/metabolismo , Desarrollo de Músculos/efectos de los fármacos , Desarrollo de Músculos/genética , Proteínas Musculares/genética , Proteínas Musculares/metabolismo , Músculo Esquelético/efectos de los fármacos , Mioblastos/efectos de los fármacos , Mioblastos/metabolismo , Nanofibras/ultraestructura , Poliésteres/farmacología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas Endogámicas Lew , Andamios del Tejido/químicaRESUMEN
Patients suffering from bacterial bloodstream infections have an increased risk of developing systematic inflammatory response syndrome (SIRS), which can result in rapid deterioration of the patients' health. Diagnostic methods for bacterial identification and antimicrobial susceptibility tests are time-consuming. The aim of this study was to investigate whether Raman spectroscopy would be able to rapidly provide an antimicrobial susceptibility profile from bacteria isolated directly from positive blood cultures. First, bacterial strains (n = 133) were inoculated in tryptic soy broth and incubated in the presence or absence of antibiotics for 5 h. Antimicrobial susceptibility profiles were analyzed by Raman spectroscopy. Subsequently, a selection of strains was isolated from blood cultures and analyzed similarly. VITEK®2 technology and broth dilution were used as the reference methods. Raman spectra from 67 antibiotic-susceptible strains showed discriminatory spectra in the absence or at low concentrations of antibiotics as compared to high antibiotic concentrations. For 66 antibiotic-resistant strains, no antimicrobial effect was observed on the bacterial Raman spectra. Full concordance with VITEK®2 data and broth dilution was obtained for the antibiotic-susceptible strains, 68 % and 98 %, respectively, for the resistant strains. Discriminative antimicrobial susceptibility testing (AST) profiles were obtained for all bacterial strains isolated from blood cultures, resulting in full concordance with the VITEK®2 data. It can be concluded that Raman spectroscopy is able to detect the antimicrobial susceptibility of bacterial species isolated from a positive blood culture bottle within 5 h. Although Raman spectroscopy is cheap and rapid, further optimization is required, to fulfill a great promise for future AST profiling technology development.
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Antibacterianos/farmacología , Bacterias/química , Bacterias/efectos de los fármacos , Cultivo de Sangre/métodos , Pruebas de Sensibilidad Microbiana/métodos , Espectrometría Raman/métodos , Bacteriemia/microbiología , Humanos , Factores de TiempoRESUMEN
Typing of methicillin-resistant Staphylococcus aureus (MRSA) remains necessary in order to assess whether transmission of MRSA occurred and to what extent infection prevention measures need to be taken. Raman spectroscopy (SpectraCellRA [SCRA]; RiverD International, Rotterdam, The Netherlands) is a recently developed tool for bacterial typing. In this study, the performance (typeability, discriminatory power, reproducibility, workflow, and costs) of the SCRA system was evaluated for typing of MRSA strains isolated from patients and patients' household members who were infected with or colonized by MRSA. We analyzed a well-documented collection of 113 MRSA strains collected from 54 households. The epidemiological relationship between the MRSA strains within one household was used as the gold standard. Pulsed-field gel electrophoresis (PFGE) was used for discrepancy analysis. The results of SCRA analysis on the strain level corresponded with epidemiological data for 108 of 113 strains, a concordance of 95.6%. When analyzed at the household level, the results of SCRA were correct for 49 out of 54 households, a concordance of 90.7%. Concordance on the strain level with epidemiological data for PFGE was 93.6% (103/110 isolates typed). Concordance on the household level with epidemiological data for PFGE was 93.5% (49/53 households analyzed). With PFGE regarded as the reference standard, the conclusions reached with Raman spectroscopy were identical to those reached with PFGE in 100 of 105 cases (95.2%). The reproducibility of SCRA was found to be 100%. We conclude that the SpectraCellRA system is a fast, easy-to-use, and highly reproducible typing platform for outbreak analysis that can compete with the currently used typing techniques.
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Técnicas de Tipificación Bacteriana , Staphylococcus aureus Resistente a Meticilina/clasificación , Staphylococcus aureus Resistente a Meticilina/aislamiento & purificación , Espectrometría Raman , Infecciones Estafilocócicas/microbiología , Brotes de Enfermedades , Electroforesis en Gel de Campo Pulsado , Humanos , Staphylococcus aureus Resistente a Meticilina/genética , Reproducibilidad de los Resultados , Infecciones Estafilocócicas/epidemiología , Infecciones Estafilocócicas/transmisiónRESUMEN
Members of the histone deacetylase (HDAC) family exhibit great promise as potential drug targets in pediatric tumors including neuroblastoma, medulloblastoma, ependymoma and Ewing's sarcoma. HDAC inhibitors of various structural classes have shown anti-tumoral effects in pre-clinical pediatric tumor models as single agents or in combination treatments. Suberoylanilidehydroxamic acid (SAHA=vorinostat) is the most clinical advanced compound of the class and was approved by the US FDA in October 2006 for the treatment of refractory cutaneous T-cell lymphoma. In this phase I/II trial, pediatric patients with relapsed solid tumors, lymphoma or leukemias are treated according to an individualized dose escalation concept ensuring each individual patient to receive his optimal dose with respect to toxicity and efficacy. The study is accompanied by an extensive pharmacokinetic, pharmacodynamic and biomarker program.
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Antineoplásicos/administración & dosificación , Inhibidores de Histona Desacetilasas/administración & dosificación , Ácidos Hidroxámicos/administración & dosificación , Leucemia/tratamiento farmacológico , Linfoma/tratamiento farmacológico , Recurrencia Local de Neoplasia/tratamiento farmacológico , Neoplasias/tratamiento farmacológico , Administración Oral , Adolescente , Antineoplásicos/farmacocinética , Niño , Preescolar , Relación Dosis-Respuesta a Droga , Esquema de Medicación , Femenino , Inhibidores de Histona Desacetilasas/farmacocinética , Humanos , Ácidos Hidroxámicos/farmacocinética , Leucemia/sangre , Cuidados a Largo Plazo , Linfoma/sangre , Masculino , Recurrencia Local de Neoplasia/sangre , Neoplasias/sangre , VorinostatRESUMEN
A broth for the screening of group B streptococcal (GBS) carriage during pregnancy is about to be introduced. Simulating conditions in everyday practice, we have compared the sensitivity of this Granada tube broth (GT) with that of classical Amies transport medium (AT) in vitro. A total of 1,485 GT and 1,485 AT were tested with 33 well-characterized GBS strains in three different concentrations, five different incubation times, and three different temperatures. After initial incubation at room temperature (RT) or 4°C, GT were placed at 37°C. GT were scored for the presence of orange pigment. GT and AT were subcultured on blood agar (BA). Pigment was observed in 98% of GT incubated at 37°C. GBS could be cultured in 91%, 73%, and 55% of GT incubated at 37°C, RT, or 4°C, respectively. For AT, these percentages were only 20% at 37°C, 52% at RT, and 59% at 4°C. When GT initially incubated at RT or 4°C were subsequently incubated at 37°C, the sensitivity improved significantly. We conclude that GT is a more sensitive GBS transport and culture medium than the conventional method, especially for low inocula and prolonged transport/incubation times. GT does not exclude the presence of GBS, and should always be incubated at 37°C and subcultured on solid agar for optimal sensitivity.
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Medios de Cultivo , Infecciones Estreptocócicas/diagnóstico , Streptococcus agalactiae/crecimiento & desarrollo , Streptococcus agalactiae/aislamiento & purificación , Femenino , Humanos , Embarazo , Complicaciones Infecciosas del Embarazo/diagnóstico , Complicaciones Infecciosas del Embarazo/microbiología , Sensibilidad y Especificidad , Infecciones Estreptocócicas/microbiologíaRESUMEN
Two multicentre external quality assessments (EQA) for the molecular detection and genotyping of meticillin-resistant Staphylococcus aureus (MRSA) were arranged. Firstly, 11 samples containing various amounts of inactivated MRSA strains, meticillin-susceptible S. aureus (MSSA), meticillin-resistant coagulase-negative staphylococci (MRCoNS) or Escherichia coli were distributed to 82 laboratories. Samples containing 102 or 103 MRSA cells were correctly scored in only 16 and 46% of the datasets returned, respectively. Two of the used MSSA strains contained an SCCmec cassette lacking the mecA gene. There was a marked difference in the percentage of correct results for these two MSSA strains (37 and 39%) compared to the MSSA strain lacking the SCCmec cassette (88%). Secondly, a panel for MRSA genotyping, consisting of ten samples (two identical, three genetically related and five unique strains) was distributed to 19 laboratories. Seventy-three percent of the datasets recorded all samples correctly. Most pulsed-field gel electrophoresis (PFGE) protocols proved to be suboptimal, resulting in inferior resolution in the higher or lower fragment regions. The performance of molecular diagnostics for MRSA shows no significant changes since our first EQA in 2006. The first molecular typing results are encouraging. Both assessments indicate that programme expansion is required and that major performance discrepancies continue to exist.
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Staphylococcus aureus Resistente a Meticilina/genética , Técnicas de Diagnóstico Molecular/métodos , Garantía de la Calidad de Atención de Salud/métodos , Infecciones Estafilocócicas/microbiología , Proteínas Bacterianas/genética , ADN Bacteriano/análisis , Electroforesis en Gel de Campo Pulsado/métodos , Genotipo , Humanos , Proteínas de Unión a las Penicilinas , Staphylococcus aureus/genéticaRESUMEN
The performances of the BDProbeTec ET (Becton Dickinson) and COBAS AMPLICOR MTB (Roche) were retrospectively evaluated for detecting Mycobacterium tuberculosis complex in various respiratory specimens. The BACTEC and MGIT liquid culture system (Becton Dickinson) was used as a reference method. A total of 824 respiratory specimens, comprised of sputa, bronchoalveolar lavage fluid, and bronchial and tracheal aspirates from 580 patients, were evaluated. Out of 824 clinical specimens, 109 specimens from 43 patients were culture positive for M. tuberculosis. Of these 109 specimens, 67 were smear positive, 85 were positive by the COBAS AMPLICOR MTB test, and 94 were positive by the BDProbeTec ET. Of the 715 culture-negative specimens, 17 were positive by the auramine staining, 11 were positive by the COBAS AMPLICOR MTB test, and 12 were positive by the BDProbeTec ET. After discrepancy analysis and review of the patients' clinical data, 130 specimens from 50 patients were considered "true-positive" specimens. This resulted in the following sensitivities: microscopy, 61.5%; COBAS AMPLICOR MTB test, 78.0%; and BDProbeTec ET, 86.2%. The specificities of each system, based on the clinical diagnosis, were 99.7% for microscopy, 99.9% for the COBAS AMPLICOR MTB test, and 99.9% for the BDProbeTec ET. The data presented represent a considerable number of specimens evaluated with a considerable number of culture- and auramine-positive and culture-positive and auramine-negative results and therefore give a realistic view of how the data should be interpreted in a daily routine situation. Specifically, the data with regard to the culture-positive and auramine-negative specimens are useful, because in a routine situation, auramine-negative specimens are sometimes accepted, on clinical indications, to be analyzed by an amplification method.
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Líquido del Lavado Bronquioalveolar/microbiología , Infecciones por Mycobacterium/diagnóstico , Mycobacterium tuberculosis/aislamiento & purificación , Esputo/microbiología , Benzofenoneido/metabolismo , Medios de Cultivo , Humanos , Infecciones por Mycobacterium/microbiología , Mycobacterium tuberculosis/clasificación , Mycobacterium tuberculosis/genética , Técnicas de Amplificación de Ácido Nucleico/instrumentación , Técnicas de Amplificación de Ácido Nucleico/métodos , Juego de Reactivos para Diagnóstico , Sensibilidad y Especificidad , Coloración y Etiquetado/métodosRESUMEN
Source images are extracted from two-particle correlations constructed from strange and nonstrange hadrons produced in 6A GeV Au+Au collisions. Very different source images result from pp vs p Lambda vs pi(-)pi(-) correlations. Scaling by transverse mass can describe the apparent source size ratio for p/pi(-) but not for Lambda/pi(-) or Lambda/p. These observations suggest important differences in the space-time emission histories for protons, pions, and neutral strange baryons produced in the same events.
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We have successfully generated and characterized a stable packaging cell line for HIV-1-based vectors. To allow safe production of vector, a minimal packaging construct carrying only the coding sequences of the HIV-1 gag-pol, tat, and rev genes was stably introduced into 293G cells under the control of a Tet(o) minimal promoter. 293G cells express the chimeric Tet(R)/VP16 trans-activator and contain a tetracycline-regulated vesicular stomatitis virus protein G (VSV-G) envelope gene. When the cells were grown in the presence of tetracycline the expression of both HIV-1-derived and VSV-derived packaging functions was suppressed. On induction, approximately 50 ng/ml/24 hr of Gag p24 equivalent of vector was obtained. After introduction of the transfer vector by serial infection, vector could be collected for several days with a transduction efficiency similar or superior to that of vector produced by transient transfection both for dividing and growth-arrested cells. The vector could be effectively concentrated to titers reaching 10(9) transducing units/ml and allowed for efficient delivery and stable expression of a GFP transgene in the mouse brain. The packaging cell line and all vector producer clones described here were shown to be free from replication-competent recombinants, and from recombinants between packaging and vector constructs that transfer the viral gag-pol genes. The packaging cell line and the assays developed will advance lentiviral vectors toward the stringent requirements of clinical applications.
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Vectores Genéticos , Lentivirus/genética , Glicoproteínas de Membrana , Animales , Antibacterianos/farmacología , Southern Blotting , Encéfalo/metabolismo , División Celular , Línea Celular , Proteínas de Fusión gag-pol/genética , Productos del Gen rev/genética , Productos del Gen tat/genética , Proteínas Fluorescentes Verdes , VIH-1/genética , Células HeLa , Humanos , Proteínas Luminiscentes/metabolismo , Ratones , Ratones Endogámicos C57BL , Modelos Genéticos , Plásmidos/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Recombinación Genética , Tetraciclina/farmacología , Factores de Tiempo , Transducción Genética , Transfección , Transgenes , Proteínas del Envoltorio Viral/genética , Productos del Gen rev del Virus de la Inmunodeficiencia Humana , Productos del Gen tat del Virus de la Inmunodeficiencia HumanaRESUMEN
Directed flow measurements for Lambda hyperons are presented and compared to those for protons produced in the same Au+Au collisions (2A, 4A, and 6A GeV; b<5-6 fm). The measurements indicate that Lambda hyperons flow consistently in the same direction but with smaller magnitudes. A strong positive flow [for Lambdas] has been predicted in calculations which include the influence of the Lambda-nucleon potential. The experimental flow ratio Lambda/p is in qualitative agreement with expectations (approximately 2/3) from the quark counting rule at 2A GeV but is found to decrease with increasing beam energy.
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Organoclorines pesticides are compounds that were utilized in agriculture until 1985, when their use was forbided, since they represented a risk for human health and environment. Nowadays, its clandestine use is still representative. These compounds may be used in public health campaigns to combat mosquitos that transmit malaria. If public health nurses know the particularities of these compounds, they will be able to work in health prevention and promotion, recognizing the chronic and acute damages as well as in the epidemiologic vigilance of the populations exposed. This paper presents a revision about the organoclorines pesticides and, based on it, proposes some activities to be implemented by nursing professionals in primary health care, aiming the health of workers exposed to the pesticides.
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Plaguicidas/envenenamiento , Enfermedad Aguda , Enfermedad Crónica , Humanos , Intoxicación/diagnóstico , Intoxicación/enfermería , Vigilancia de la Población , Salud RuralRESUMEN
This paper describes the experience of a teacher and four students of the Federal University of Rio Grande do Sul within a sitting of members of a rural social movement that claims for land in Brazil. The work began with an activity of familiarization and ended up being an extension project. The aim of this project was the development of a Nursing assistencial model for the population of the rural area, considering the specificities of this movement, wishing to adapt Nursing teaching to this reality. The author sought for theoretical references in Breilh (1999) and Freire (1981). The activities were developed in 1996 and consisted of home care and group education.
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Servicios de Enfermería , Servicios de Salud Rural , Brasil , Humanos , Factores SocioeconómicosRESUMEN
We studied the prevalence and clonality of high-level gentamicin-resistant enterococci (HLGRE) in a Dutch university hospital. Of 238 enterococcal strains isolated from blood cultures between 1991 and 1997, 57 were HLGRE. Genomic analysis of these strains revealed 19 different genotypes, two of which were encountered more frequently [type A (12/57), type B (23/57)]. The spread of these types largely explained the rise in HLGRE incidence from 14% in 1991 to 31% in 1997. However, the contribution of unique strains to the total HLGRE burden also increased from 4% to 16%. We conclude that both clonal expansion and the emergence of unique HLGRE have contributed significantly to the increasing incidence of HLGRE.
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Antibacterianos/farmacología , Enterococcus/efectos de los fármacos , Enterococcus/genética , Gentamicinas/farmacología , Infecciones por Bacterias Grampositivas/epidemiología , Infecciones por Bacterias Grampositivas/microbiología , Bacteriemia/epidemiología , Bacteriemia/microbiología , Sangre/microbiología , Medios de Cultivo , ADN Bacteriano/análisis , Farmacorresistencia Microbiana/genética , Electroforesis en Gel de Campo Pulsado , Enterococcus/aislamiento & purificación , Hospitales Universitarios , Humanos , Pruebas de Sensibilidad Microbiana , Países Bajos/epidemiología , Reacción en Cadena de la Polimerasa/métodos , PrevalenciaRESUMEN
Permanent facial nerve dysfunction is a potential complication of every parotid surgery. Partial superficial parotidectomy, a conservative resectioning that requires neither dissection of the full facial nerve nor excision of the superficial lobe, produces lower rates of facial nerve dysfunction and soft tissue deformity than the traditional method. This report describes a single surgeon's experience with partial superficial parotidectomy from 1987 to 1997. Fifty-nine patients with mobile, benign, and low-grade malignant tumors, limited to the superficial lobe, underwent partial superficial parotidectomy with selective nerve dissection. Adequate margins were obtained, based on the premise that the tumor-to-nerve margin is often the true one. No patients had permanent nerve paralysis or paresis, and only 10 incurred transient facial nerve paresis. Age, histology, and sex were not significant factors in postoperative facial nerve function. No patients had recurrences.