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1.
J Hosp Infect ; 145: 142-147, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38272124

RESUMO

BACKGROUND: A recent study confirmed significant contamination of syringe tips following routine anaesthesia practice of at least 6 h in duration. AIM: We assessed the relative efficacy of clinically relevant syringe tip disinfection techniques following contamination with the hyper transmissible and more pathogenic Staphylococcus aureus sequence type 5 (S. aureus ST5) strain characteristic associated with increased strength of biofilm formation and greater desiccation tolerance. METHODS: Syringe tips (N=40) contaminated with S. aureus ST5 were randomized to 70% isopropyl pads with 10 or 60 s of drying time, scrubbing alcohol disinfection caps with 10 or 60 s of dwell time, or to non-scrubbing alcohol disinfection caps with 60 s of dwell time. The primary outcome was residual 24-h colony forming units (cfu) >10. RESULTS: Scrubbing disinfection caps were more effective than alcohol pads (25% (12/48) <10 cfu for scrubbing caps (10- or 60-s dwell times) vs 0% (0/48) <10 cfu for alcohol pads (10 or 60 s of drying time), Holm-Sidak adjusted P=0.0016). Scrubbing disinfection caps were more effective than non-scrubbing alcohol disinfection caps (25% (12/48) <10 cfu for scrubbing alcohol caps (10- or 60-s dwell times) vs 2% (1/48) for non-scrubbing alcohol caps (60-s dwell time), adjusted P=0.0087). CONCLUSIONS: Scrubbing alcohol caps are more effective than alcohol pads or non-scrubbing disinfecting caps for microbial reduction of syringe tips contaminated with the more pathogenic S. aureus ST5.


Assuntos
Desinfecção , Staphylococcus , Humanos , Desinfecção/métodos , Staphylococcus aureus , Seringas , Etanol , Contaminação de Equipamentos
2.
J Hosp Infect ; 143: 186-194, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37451409

RESUMO

BACKGROUND: Staphylococcus aureus sequence type 5 (ST5) is an emerging global threat. AIM: To characterize the epidemiology of ST5 transmission in the anaesthesia work area. METHODS: The retrospective cohort study analysed transmitted, prophylactic antibiotic-resistant Staphylococcus aureus isolates involving anaesthesia work area reservoirs. Using whole-genome analysis, the epidemiology of ST5 transmission was characterized by reservoir(s) of origin, transmission location(s), portal of entry, and mode(s) of transmission. All patients were followed for at least 30 days for surgical site infection (SSI) development. FINDINGS: Forty-one percent (18/44; 95% confidence interval: 28-56%) of isolates were ST5. Provider hands were the reservoir of origin for 28% (5/18) of transmitted ST5 vs 4% (1/26) for other STs. Provider hands were the transmission location for 28% (5/18) of ST5 vs 7% (2/26) of other STs. Stopcock contamination occurred for 8% (1/13) of ST5 isolates vs 12% (3/25) of other STs. Sixty-three percent of transmission events occurring between cases on separate operative dates involved ST5. ST5 was more likely to harbour resistance traits (ST5 median (interquartile range) 3 (2-3) vs 2 (1-2) other STs; P < 0.001) and had greater resistance to cefazolin, piperacillin-tazobactam, and/or ciprofloxacin (ST5: 3 (2-3) vs 2 (1-3) other STs; P = 0.02). ST5 was associated with three of six SSIs. CONCLUSION: ST5 is prevalent among transmitted, prophylactic antibiotic-resistant isolates in the anaesthesia work area. Transmission involves provider hands and one patient to another on future date(s). ST5 is associated with a greater number of resistance traits and reduced in-vitro susceptibility vs other intraoperative meticillin-resistant S. aureus.


Assuntos
Anestesia , Staphylococcus aureus Resistente à Meticilina , Infecções Estafilocócicas , Humanos , Staphylococcus aureus/genética , Epidemiologia Molecular , Estudos Retrospectivos , Infecções Estafilocócicas/prevenção & controle , Antibacterianos/farmacologia , Testes de Sensibilidade Microbiana
3.
J Hosp Infect ; 134: 121-128, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36693592

RESUMO

BACKGROUND: The extent to which the transmission of prophylactic-antibiotic-resistant bacteria from the anaesthesia work area increases the risk of surgical site infection (SSI) is unknown. It was hypothesized that the risk of SSI would increase progressively from no transmission to transmission of prophylactic-antibiotic-resistant isolates. METHODS: This was a retrospective analysis of archival samples collected in two previously published studies with similar inclusion criteria and sample collection methodology (observational study 2009-2010 and randomized trial 2018-2019). Archival isolates were linked by barcode to all patient demographic and procedural information, including the prophylactic antibiotic administered, transmission and development of SSI. For this study, all archival isolates underwent prophylactic antibiotic susceptibility testing, and the ordered association of transmission of Staphylococcus aureus (no transmission, transmission of prophylactic-antibiotic-susceptible isolates and transmission of prophylactic-antibiotic-resistant isolates) with SSI was assessed. RESULTS: The risk of development of SSI was 2% (8/406) without S. aureus transmission, 11% (9/84) with transmission of S. aureus isolates that were susceptible to the prophylactic antibiotic used, and 18% (4/22) with transmission of prophylactic-antibiotic-resistant S. aureus isolates. The Cochrane-Armitage two-sided test for ordered association was P<0.0001. Treating these three groups as 0, 1 and 2, by exact logistic regression, the odds of SSI increased by 3.59 with each unit increase (95% confidence interval 1.92-6.64; P<0.0001). CONCLUSIONS: Transmission of S. aureus in the anaesthesia work area reliably increases the risk of SSI, especially when the isolates are resistant to the prophylactic antibiotic administered.


Assuntos
Anestesia , Staphylococcus aureus Resistente à Meticilina , Infecções Estafilocócicas , Humanos , Staphylococcus aureus , Infecção da Ferida Cirúrgica/epidemiologia , Infecção da Ferida Cirúrgica/prevenção & controle , Infecção da Ferida Cirúrgica/tratamento farmacológico , Estudos Retrospectivos , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/prevenção & controle , Infecções Estafilocócicas/tratamento farmacológico , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico
4.
Artigo em Inglês | MEDLINE | ID: mdl-37359471

RESUMO

Background: Rapid sequence intubation (RSI) involves inducing unconsciousness and paralysis in rapid succession in order to facilitate endotracheal tube placement. RSI has recently been introduced to the scope of practice of South African prehospital emergency care practitioners (ECPs). Despite this, there remains limited evidence supporting the efficacy and safety of RSI within this context. While in-hospital studies have shown that it can take 20 minutes or more to perform an RSI, little is known about the time taken to perform the procedure in the prehospital setting. Objectives: To measure the time taken to perform an RSI in a simulated prehospital environment. Methods: A sample of final-year ECP students were video-recorded performing RSIs on a mannequin within a simulated prehospital environment. Data were gathered through an analysis of the recordings, allowing for the capturing of times taken to complete each of the phases of a RSI. Results: A mean time of 15 minutes 5 seconds was recorded to complete the procedure. This was shorter than times reported for in-hospital studies. Conclusion: RSI is a potentially harmful procedure if improperly performed and has the potential to create delays in transport that may not always be in the patient's best interest. With a mean time of 15 minutes 5 seconds, the performance of RSI by ECP students in the simulated prehospital environment was faster than expected. Further research is recommended to explore the relationship between the performances observed in this mannequin-based study with those in authentic prehospital settings. Contributions of the study: This study adds to a currently limited body of knowledge surrounding the performance of out-of-hospital anaesthesia by emergency care practitioners in the African context. The study highlights the fact that while prehospital rapid sequence intubation may be a lifesaving procedure, anaesthetising patients in an uncontrolled prehospital environment is not without risk. An important consideration that needs to be taken into account when making a decision on whether or not to perform the procedure within the prehospital setting is the potential delay this might have on transport time and arrival at the receiving facility.

5.
J Hosp Infect ; 100(3): 299-308, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29966756

RESUMO

BACKGROUND: Desiccation tolerance increases Staphylococcus aureus survival and risk of transmission. A better understanding of factors driving intraoperative transmission of S. aureus pathogens may lead to innovative improvements in intraoperative infection control. AIMS: To determine whether desiccation tolerance is associated with intraoperative S. aureus transmission, and to examine typical transmission dynamics for desiccation-tolerant isolates in the operating room in order to provide the impetus for development of improved intraoperative infection control strategies. METHODS: S. aureus isolates (N=173) were collected from anaesthesia work area reservoirs in 274 operating room environments. Desiccation tolerance was assessed and the potential association with sequence type (ST) and clonal transmission was evaluated. Whole cell genome analysis and pulsed-field gel electrophoresis analysis were used to compare desiccation-tolerant isolates with causative organisms of infection. FINDINGS: S. aureus ST 5 isolates had greater desiccation tolerance than all other intraoperative STs [ST 5, N=34, median Day 2 colony-forming unit (cfu) survival 0.027% ± 0.029%; other STs, N=139, median Day 2 cfu survival 0.0091% ± 1.41%; corrected P=0.0001]. ST 5 was associated with increased risk of clonal transmission (relative risk 1.82, 95% confidence interval 1.23-2.71, P=0.003). ST 5 transmission was linked by whole cell genome analysis to postoperative infection. CONCLUSIONS: Increased desiccation tolerance is associated with intraoperative transmission of S. aureus ST 5 isolates that are linked to postoperative infection. Future work should determine whether attenuation of desiccation-tolerant, intraoperative ST 5 strains can impact the incidence of healthcare-associated infections.


Assuntos
Desidratação , Transmissão de Doença Infecciosa , Viabilidade Microbiana , Salas Cirúrgicas , Infecções Estafilocócicas/transmissão , Staphylococcus aureus/isolamento & purificação , Staphylococcus aureus/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Eletroforese em Gel de Campo Pulsado , Microbiologia Ambiental , Feminino , Genótipo , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Tipagem de Sequências Multilocus , Staphylococcus aureus/classificação , Staphylococcus aureus/genética , Sequenciamento Completo do Genoma
6.
Rev Sci Tech ; 24(1): 231-42, 2005 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-16110891

RESUMO

Traceability is increasingly becoming standard across the agri-food industry, largely driven by recent food crises and the consequent demands for transparency within the food chain. This is leading to the development of a range of traceability concepts and technologies adapted to different industry needs. Experience with genetically modified plants has shown that traceability can play a role in increasing public confidence in biotechnology, and might similarly help allay concerns relating to the development of animal biotechnology. Traceability also forms an essential component of any risk management strategy and is a key requirement for post-marketing surveillance. Given the diversity of traceability concepts and technologies available, consideration needs to be given to the scope and precision of traceability systems for animal biotechnology. Experience to date has shown that conventional tagging and labelling systems can incorporate levels of error and may not have sufficient precision for biotech-derived animals. Deoxyribonucleic acid (DNA) technology can overcome these difficulties by tracing animals and animal by-products through their DNA code rather than an associated label. This offers the possibility of tracing some by-products of animal biotechnology through the supply chain back to source animals, offering unprecedented levels of traceability. Developments in both DNA sampling and analysis technology are making large-scale applications of DNA traceability increasingly cost effective and feasible, and are likely to lead to a broader uptake of DNA traceability concepts.


Assuntos
Sistemas de Identificação Animal/veterinária , Animais Geneticamente Modificados , Marcadores Genéticos , Vacinas de DNA , Sistemas de Identificação Animal/métodos , Animais , Alimentos Geneticamente Modificados , Engenharia Genética , Vigilância de Produtos Comercializados
7.
Mol Ecol ; 13(11): 3477-87, 2004 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-15488005

RESUMO

We present a population genetic analysis of microsatellite variation in 16 West African cattle populations. West Africa represents a unique juxtaposition of different climatic and ecological zones in a relatively small geographical area. While more humid coastal regions are inhabited by the tsetse fly, a vector which spreads trypanosomiasis among cattle, the disease is not transmitted in the drier areas outside this zone. This is the most thorough study of genetic diversity in cattle within this area, which contains genetically important trypanotolerant Bos taurus breeds. Genetic relationships among the many breeds are examined and levels of diversity are assessed. Admixture levels were determined using a variety of methods. Ancestry informative or population-associated alleles (PAAs) were selected using populations from India, the Near East and Europe. Multivariate analysis, the admix program and model-based Bayesian admixture analysis approaches were also employed. These analyses reveal the direct impact of ecological factors and the profound effect of admixture on the cattle of this region. They also highlight the importance of efforts to prevent further dilution of African taurine breeds by B. indicus cattle.


Assuntos
Bovinos/genética , Variação Genética , África , Animais , Cruzamento , Bovinos/classificação , Ecossistema , Imunidade Inata/genética , Repetições de Microssatélites , Análise Multivariada , Filogenia , Tripanossomíase Bovina/transmissão
8.
Heredity (Edinb) ; 91(1): 43-50, 2003 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-12815452

RESUMO

We present population genetic analysis of microsatellite variation in seven Bos indicus cattle breeds from a variety of locations in South Asia. This is the first such study focusing within this area, which is one of the postulated centres of origin of domestic cattle. An estimate of the influence of Bos taurus ancestry was carried out using three approaches: by the systematic selection of population-associated alleles for B. taurus and examination of their frequency; by examining the truncation of genetic distances from European populations; and by a model-based Bayesian admixture analysis. These analyses revealed a B. taurus influence in the Indian subcontinent; part of a gradation which stretches from Europe through the Near East towards Indian and which may be of ancient origin.


Assuntos
Bovinos/genética , Variação Genética , Repetições de Microssatélites/genética , Alelos , Animais , Teorema de Bayes , Genética Populacional , Índia , Paquistão
9.
Nature ; 410(6832): 1088-91, 2001 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-11323670

RESUMO

The limited ranges of the wild progenitors of many of the primary European domestic species point to their origins further east in Anatolia or the fertile crescent. The wild ox (Bos primigenius), however, ranged widely and it is unknown whether it was domesticated within Europe as one feature of a local contribution to the farming economy. Here we examine mitochondrial DNA control-region sequence variation from 392 extant animals sampled from Europe, Africa and the Near East, and compare this with data from four extinct British wild oxen. The ancient sequences cluster tightly in a phylogenetic analysis and are clearly distinct from modern cattle. Network analysis of modern Bos taurus identifies four star-like clusters of haplotypes, with intra-cluster diversities that approximate to that expected from the time depth of domestic history. Notably, one of these clusters predominates in Europe and is one of three encountered at substantial frequency in the Near East. In contrast, African diversity is almost exclusively composed of a separate haplogroup, which is encountered only rarely elsewhere. These data provide strong support for a derived Near-Eastern origin for European cattle.


Assuntos
Bovinos/genética , África , Animais , Animais Selvagens , Bovinos/classificação , DNA Mitocondrial , Europa (Continente) , Variação Genética , Haplótipos , Oriente Médio , Dados de Sequência Molecular , Filogenia , Ruminantes/classificação , Ruminantes/genética
10.
Mol Endocrinol ; 15(4): 575-88, 2001 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-11266509

RESUMO

Aldosterone stimulates Na(+) reabsorption in the collecting ducts by increasing the activity of the epithelial sodium channel, ENaC. Systemic administration of aldosterone increases alpha ENaC mRNA expression in mammalian kidney, suggesting that the alpha ENaC gene is a target for aldosterone action in the distal nephron. To determine whether aldosterone increases alpha ENaC gene transcription, a portion of the alpha ENaC 5'- flanking region coupled to luciferase was transfected into MDCK-C7 cells, a collecting duct cell line with aldosterone-stimulated Na(+) transport. Both dexamethasone and aldosterone stimulated alpha ENaC-coupled reporter gene activity via the glucocorticoid receptor (GR), and this response correlated with the effect of these hormones on endogenous alpha ENaC expression. The aldosterone-stimulated alpha ENaC expression was blocked by actinomycin D, and aldosterone had no effect on alpha ENaC mRNA decay, confirming a transcriptional effect. In HT-29 cells, a GR/mineralocorticoid receptor (MR)-deficient colonic cell line with constitutive alpha ENaC expression, cotransfection with GR or MR restored aldosterone-stimulated alpha ENaC gene transcription, although aldosterone had a functional preference for MR. Analysis of deletion constructs confirmed that a single imperfect glucocorticoid response element (GRE) is necessary and sufficient to confer the aldosterone responsiveness to the alpha ENaC gene promoter in MDCK-C7 and HT-29 cells. These results confirm that alpha ENaC is an aldosterone-induced transcript in the collecting duct and delineates the molecular mechanism for this effect.


Assuntos
Aldosterona/metabolismo , Túbulos Renais Coletores/fisiologia , Sequências Reguladoras de Ácido Nucleico , Canais de Sódio/genética , Transcrição Gênica , Aldosterona/farmacologia , Animais , Sequência de Bases , Sítios de Ligação , Células Cultivadas , Clonagem Molecular , Dactinomicina/farmacologia , Dexametasona/farmacologia , Cães , Canais Epiteliais de Sódio , Gonanos/farmacologia , Humanos , Túbulos Renais Coletores/citologia , Camundongos , Mifepristona/farmacologia , Dados de Sequência Molecular , Regiões Promotoras Genéticas , Subunidades Proteicas , Receptores de Glucocorticoides/antagonistas & inibidores , Receptores de Glucocorticoides/efeitos dos fármacos , Receptores de Glucocorticoides/metabolismo , Receptores de Mineralocorticoides/efeitos dos fármacos , Receptores de Mineralocorticoides/metabolismo , Elementos de Resposta , Canais de Sódio/metabolismo
11.
Anim Genet ; 31(5): 329-32, 2000 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-11105215

RESUMO

We estimated the genetic relationships between the endangered German Pustertaler-Sprinzen cattle breed and the Pinzgauer, Vosges and Simmental breeds--decided upon after consultation of the available historical literature. Within-breed diversity of the four breeds was also assessed. Twenty microsatellite markers were amplified in 27-50 unrelated individuals from populations of each breed. Within-breed variation was estimated from average heterozygosity values and mean number of alleles. Breed relationships were evaluated by genetic distance and a neighbour-joining tree was calculated from these estimates. Bootstrap resampling of loci tested the robustness of the tree topology obtained. A tree was also constructed from distance matrices using individual animals as operational taxonomic units. From both the average heterozygosity values and mean number of alleles calculated, the Pustertaler breed appears to be no more genetically impoverished than the other breeds analysed. The breed tree showed an 85% support for the Pustertaler-Pinzgauer grouping, and this result is echoed in the genetic distance values and allele-sharing individual tree.


Assuntos
Bovinos/genética , Variação Genética , Repetições de Microssatélites , Animais , Bovinos/classificação , Conservação dos Recursos Naturais , Europa (Continente) , Triagem de Portadores Genéticos , Marcadores Genéticos , Genótipo
12.
Biochem J ; 347 Pt 1: 105-14, 2000 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-10727408

RESUMO

The mRNA for the epithelial Na(+) channel gamma subunit (gammaENaC) is regulated developmentally in the lung, colon and distal nephron and in response to Na(+) deprivation and systemic corticosteroids in the distal colon. Because such regulation is likely to be at the level of gene transcription, we examined the function of the promoter and other 5' flanking elements of the human gammaENaC gene. The proximal 5' flanking region contains two GC boxes but does not contain a TATA box. A 450 bp human gammaENaC fragment (-459 to +40) directed the expression of luciferase in H441 cells and primer extension analysis in transfected cells confirmed the correct initiation of human gammaENaC-luciferase chimaeric transcripts. By deletional analysis, GC boxes at -21 and -52 were found to be critical for this promoter activity. To begin to identify transcription factors that bind to the core promoter, a double-stranded oligonucleotide that corresponded to this region was synthesized and tested in a gel mobility-shift assay. Incubation of this radiolabelled oligonucleotide with nuclear extracts from H441 and FRTL5 cells resulted in the formation of four specific and distinct DNA-protein complexes. On the basis of antibody 'supershift' assays, one of these factors corresponds to Sp1, whereas the other three correspond to Sp3. Further upstream, an approx. 300 nt (-1143 to -839) polypurine-polypyrimidine tract (PPy tract) containing internal mirror repeats was identified. When contained in a supercoiled plasmid, the approx. 1200 nt 5' flanking region was sensitive to S1 endonuclease, which was consistent with the formation of an intramolecular triplex DNA ('H-DNA') structure with an unpaired single strand. High-resolution mapping with S1 endonuclease and sequencing of S1-generated clones confirmed that all S1-sensitive sites were within the PPy tract. Finally, a negative regulatory element was identified between -1525 and -1296 that functioned in lung, colon and collecting duct cell lines.


Assuntos
Regiões 5' não Traduzidas/genética , DNA/química , DNA/genética , Regiões Promotoras Genéticas , Canais de Sódio/genética , Sequência de Bases , Primers do DNA , Canais Epiteliais de Sódio , Humanos , Substâncias Macromoleculares , Modelos Moleculares , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Proteínas Recombinantes de Fusão/biossíntese , Sequências Repetitivas de Ácido Nucleico , Mapeamento por Restrição , Alinhamento de Sequência , Transcrição Gênica
13.
AIDS ; 13(6): F35-43, 1999 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-10397555

RESUMO

OBJECTIVE: To determine the HIV RNA and CD4 cell response to both initial and salvage therapy with protease inhibitor-based therapy, and to examine the relationship between the virological response and pre-therapy characteristics. DESIGN: Observational cohort. SETTING: University-based public hospital AIDS clinic. PATIENTS: HIV-infected adults who received at least 16 continuous weeks' therapy with a potent protease inhibitor (indinavir, ritonavir or nelfinavir)-based regimen, and who have had at least 48 weeks of follow-up. MAIN OUTCOME MEASURES: Plasma HIV RNA and CD4 cell count response at week 48 of therapy for patients receiving their first protease inhibitor-containing regimen, and at week 24 of therapy with a salvage regimen. RESULTS: Of the 337 patients analysed, 170 (50.2%) had a successful outcome (HIV RNA <500 copies/ml after 48 weeks of treatment). Independent predictors of virological failure were higher baseline HIV RNA level, lower baseline CD4 cell count and failure to initiate at least one new nucleoside analog simultaneously at the time protease inhibitor therapy was initiated. The risk of failure increased incrementally across most HIV RNA and CD4 cell strata, with significant increases as the HIV RNA increased above 4.5 log10 copies/ml and the CD4 cell count fell below 100 cells/mm3 (P< or =0.01). The CD4 cell count remained above baseline to week 48 in most patients, regardless of the HIV RNA response. Of the 99 patients who experienced virological failure and switched to a salvage regimen, only 22 (22%) achieved an undetectable HIV RNA level 24 weeks after initiating salvage therapy. Independent predictors of failure with salvage therapy included an HIV RNA greater than 4.0 log10 RNA copies/ml at the time of the switch and failure to use a non-nucleoside reverse transcriptase inhibitor (NNRTI) in the salvage regimen. CONCLUSION: Failure of potent protease inhibitor therapy to suppress HIV RNA levels below detectable levels is common in clinical practice, and can often be explained by their suboptimal use. CD4 T cell counts remain above baseline for at least one year in most patients experiencing virological failure. Successful salvage therapy, which was uncommon, was associated with a low plasma HIV RNA at the time of the switch and the use of a new class of antiretroviral agents (NNRTI) in the salvage regimen.


Assuntos
Contagem de Linfócito CD4 , Infecções por HIV/tratamento farmacológico , Inibidores da Protease de HIV/uso terapêutico , HIV-1/genética , RNA Viral/sangue , Adulto , Instituições de Assistência Ambulatorial , Quimioterapia Combinada , Infecções por HIV/sangue , Hospitais Universitários , Humanos , Resultado do Tratamento , População Urbana , Carga Viral
14.
J Biol Chem ; 274(18): 12431-7, 1999 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-10212217

RESUMO

In airway and renal epithelia, the glucocorticoid-mediated stimulation of amiloride-sensitive Na+ transport is associated with increased expression of the epithelial Na+ channel alpha subunit (alphaENaC). In H441 lung cells, 100 nM dexamethasone increases amiloride-sensitive short-circuit current (3.3 microA/cm2 to 7.5 microA/cm2), correlating with a 5-fold increase in alphaENaC mRNA expression that could be blocked by actinomycin D. To explore transcriptional regulation of alphaENaC, the human alphaENaC 5'-flanking region was cloned and tested in H441 cells. By deletion analysis, a approximately 150-base pair region 5' to the upstream promoter was identified that, when stimulated with 100 nM dexamethasone, increased luciferase expression 15-fold. This region, which contains two imperfect GREs, also functioned when coupled to a heterologous promoter. When individually tested, only the downstream GRE functioned in cis and bound GR in a gel mobility shift assay. In the M-1 collecting duct line Na+ transport, malphaENaC expression and luciferase expression from alphaENaC genomic fragments were also increased by 100 nM dexamethasone. In a colonic cell line, HT29, trans-activation via a heterologously expressed glucocorticoid receptor restored glucocorticoid-stimulated alphaENaC gene transcription. We conclude that glucocorticoids stimulate alphaENaC expression in kidney and lung via activation of a hormone response element in the 5'-flanking region of halphaENaC and this response, in part, is the likely basis for the up-regulation of Na+ transport in these sites.


Assuntos
Dexametasona/farmacologia , Córtex Renal/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Canais de Sódio/genética , Ativação Transcricional/efeitos dos fármacos , Amilorida/farmacologia , Sequência de Bases , Linhagem Celular , Primers do DNA , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Canais Epiteliais de Sódio , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Transporte de Íons , Córtex Renal/citologia , Córtex Renal/metabolismo , Pulmão/citologia , Pulmão/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Glucocorticoides/metabolismo , Células Tumorais Cultivadas
15.
Proc Biol Sci ; 266(1419): 597-603, 1999 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-10212450

RESUMO

A total of 49 samples from indigenous Portuguese cattle breeds were analysed for sequence variation in the hypervariable region of the mitochondrial DNA D-loop. Sequence comparison and phylogenetic analyses revealed that haplotypes fell into two distinct groups. These corresponded with two separate haplotype clusters into which, respectively, all African, or alternatively all sequences of European origin, have previously been shown to fall. Here, the majority of sequences of African type were encountered in three southern, as compared to three northern breeds. This pattern of African influence may reflect an intercontinental admixture in the initial origins of Iberian breeds, or it is perhaps an introgression dating from the long and influential Moorish occupation of the south of the Iberian peninsula.


Assuntos
Bovinos/genética , DNA Mitocondrial/genética , África , Animais , Sequência de Bases , Primers do DNA/genética , Variação Genética , Genética Populacional , Haplótipos , Filogenia , Portugal
16.
Hum Biol ; 71(1): 27-42, 1999 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-9972096

RESUMO

Two minisatellite (D1S80, D17S5) and 10 microsatellite (D2S1328, TPO, D3S1358, D9S926, D11S2010, THO1, VWF, FES, D16S310, and D18S848) polymorphic loci were analyzed in 5 Greek population groups (eastern Macedonia, central Macedonia, Thessaly, Epirus, and Greeks from Asia Minor) using the polymerase chain reaction. The genotypes at these loci conformed to Hardy-Weinberg equilibrium, and pairwise comparisons between them were in agreement with the expectation of independence between loci. This along with the low values of the coefficient of gene differentiation (GST) and the high heterozygosity levels of all loci allows the use of allele frequency data from the 12 hypervariable DNA markers for medicolegal casework in the Greek population groups studied. The small genetic distances indicate a genetic affinity among the 5 population samples. However, a few markers seem to allow some discrimination among the groups. No significant differences with other European populations were found for the loci studied.


Assuntos
DNA Satélite/análise , Frequência do Gene , Repetições de Microssatélites/genética , Repetições Minissatélites/genética , Polimorfismo Genético , Sequência de Bases , Distribuição de Qui-Quadrado , Feminino , Ligação Genética , Grécia , Humanos , Masculino , Dados de Sequência Molecular , Análise Multivariada , Reação em Cadeia da Polimerase , Estudos de Amostragem
17.
Mol Ecol ; 8(12): 2015-22, 1999 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-10632853

RESUMO

Eight humpless cattle breeds from the Near East, three from Europe, one from West Africa and two zebu breeds from India were screened with 20 microsatellite loci. Breeds from the Near East revealed considerable levels of introgression from zebu cattle, which was apparent most in populations from the East and which declined in populations further West. This nonrandom pattern is suggestive of the introduction of zebu cattle from the East. Notwithstanding the overlay of zebu alleles, it was possible to demonstrate that Near Eastern cattle exhibited significantly higher levels of allelic diversity than breeds from other regions, which is consistent with the view that this region represents a primary domestication centre for Bos taurus cattle. The hypothesis that B. taurus and B. indicus cattle have separate domestic origins is also supported by the survey, a large genetic divergence being apparent between the nonhybrid taurine and zebu groups.


Assuntos
Bovinos/genética , Variação Genética , Genética Populacional , Repetições de Microssatélites , África , Animais , Cruzamento , Europa (Continente) , Índia
18.
Genetics ; 150(3): 1169-75, 1998 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-9799268

RESUMO

This article describes complete mitochondrial DNA displacement loop sequences from 32 Japanese Black cattle and the analysis of these data in conjunction with previously published sequences from African, European, and Indian subjects. The origins of North East Asian domesticated cattle are unclear. The earliest domestic cattle in the region were Bos taurus and may have been domesticated from local wild cattle (aurochsen; B. primigenius), or perhaps had an origin in migrants from the early domestic center of the Near East. In phylogenetic analyses, taurine sequences form a dense tree with a center consisting of intermingled European and Japanese sequences with one group of Japanese and another of all African sequences, each forming distinct clusters at extremes of the phylogeny. This topology and calibrated levels of sequence divergence suggest that the clusters may represent three different strains of ancestral aurochs, adopted at geographically and temporally separate stages of the domestication process. Unlike Africa, half of Japanese cattle sequences are topologically intermingled with the European variants. This suggests an interchange of variants that may be ancient, perhaps a legacy of the first introduction of domesticates to East Asia.


Assuntos
Evolução Biológica , DNA Mitocondrial/genética , Variação Genética , Animais , Bison/genética , Bovinos
19.
Anim Genet ; 29(5): 333-40, 1998 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9800321

RESUMO

Genotype data from 20 microsatellites typed in 253 animals is used here to assess the genetic structure of seven European pedigree cattle breeds. Estimation of genetic subdivision using classical drift-based measures shows that the average proportion of genetic variation among breeds varies between 10 and 11% of the total, depending on the estimator used. We demonstrate that a simple allele-sharing genetic distance parameter can be used to construct a dendrogram of relationships among animals. This phylogenetic tree displays a remarkable degree of breed clustering and reflects an extensive underlying kinship structure, particularly for the Swiss Simmental breed and four breeds originating from the British Isles. Condensation of allele frequencies and individual genotypic compositions using principal component analysis is also used to investigate genetic structure among breeds and individual animals. In addition, the underlying genetic demarcation of European cattle breeds is emphasized in simulations of breeds assignment using allele frequency distributions from samples of microsatellite loci. Correct breed designation can be inferred with accuracies approaching 100% using data from a panel of 10 microsatellite loci.


Assuntos
Bovinos/genética , Repetições de Microssatélites , Alelos , Animais , Bovinos/classificação , Europa (Continente) , Feminino , Frequência do Gene , Variação Genética , Genótipo , Masculino , Análise Multivariada , Filogenia , Especificidade da Espécie
20.
J Clin Invest ; 101(11): 2301-8, 1998 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-9616201

RESUMO

The thymus in adults infected with the HIV-1 is generally thought to be inactive, both because of age-related involution and viral destruction. We have revisited the question of thymic function in adults, using chest-computed tomography (CT) to measure thymic tissue in HIV-1-seropositive (n = 99) or HIV-1-seronegative (n = 32) subjects, and correlating these results with the level of circulating CD4(+) and CD8(+) T cells that are phenotypically described as naive thymic emigrants. Abundant thymic tissue was detectable in many (47/99) HIV-1-seropositive adults, aged 20-59. Independent of age, radiographic demonstration of thymic tissue was significantly associated with both a higher CD4(+) T cell count (P = 0.02) and a higher percentage and absolute number of circulating naive (CD45RA+CD62L+) CD4(+) T cells (P < 0.04). The prevalence of an abundant thymus was especially high in younger HIV-1-seropositive adults ( 40 yr) regardless of CD4 count (P = 0.03). These studies suggest that the thymus is functional in some but not all adults with HIV-1 disease.


Assuntos
Síndrome da Imunodeficiência Adquirida/imunologia , HIV-1 , Timo/fisiopatologia , Adulto , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Estudos Transversais , Humanos , Pessoa de Meia-Idade , Tomografia Computadorizada por Raios X
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