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1.
Clin Med (Lond) ; 24(4): 100226, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38971373

RESUMO

We describe the case of a male heavy machinery operator who presented from work with a rapidly evolving spinal cord syndrome. Spinal MRI revealed thoracic vertebral body and cord infarction and evolving mild disc prolapse attributed to fibrocartilaginous disc embolism (FCDE). FCDE should be considered as one of the aetiological mechanisms of acute spinal cord infarction in pile-driver/heavy machinery operators, especially in association with adjacent vertebral body infarction and intervertebral disc prolapse. Magnetic resonance imaging (MRI) changes may evolve, warranting early follow-up MRI in appropriate cases.


Assuntos
Embolia , Infarto , Imageamento por Ressonância Magnética , Medula Espinal , Humanos , Masculino , Infarto/diagnóstico por imagem , Infarto/etiologia , Embolia/diagnóstico por imagem , Embolia/diagnóstico , Embolia/etiologia , Medula Espinal/diagnóstico por imagem , Medula Espinal/irrigação sanguínea , Medula Espinal/patologia , Corpo Vertebral/diagnóstico por imagem , Adulto , Vértebras Torácicas/diagnóstico por imagem , Doenças das Cartilagens/diagnóstico por imagem , Deslocamento do Disco Intervertebral/diagnóstico por imagem , Deslocamento do Disco Intervertebral/complicações
2.
J Neurol Sci ; 441: 120334, 2022 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-36030623

RESUMO

BACKGROUND: Data are limited on the ability of dipyridamole to additionally inhibit platelet function/reactivity in ischaemic cerebrovascular disease (CVD) patients on aspirin. AIMS: To assess inhibition of platelet function/reactivity and platelet activation with dipyridamole in CVD. METHODS: This prospective, observational study assessed TIA/ischaemic stroke patients before (baseline; N = 60), at 14 ±7 days (14d, N = 39) and ≥ 90 days (90d, N = 31) after adding dipyridamole to aspirin. Platelet function/reactivity at high shear stress (PFA-100® C-ADP) and low shear stress (VerifyNow® P2Y12 and Multiplate® ADP assays), and platelet activation status (% expression of CD62P, CD63 and leucocyte-platelet complexes on whole blood flow cytometry) were quantified. 'Dipyridamole-high on-treatment platelet reactivity (HTPR)' was defined as failure to inhibit ADP-induced platelet aggregation +/- adhesion compared with the patient's baseline on aspirin monotherapy by more than twice the coefficient-of-variation of the assay after adding dipyridamole to aspirin. RESULTS: Dipyridamole-HTPR was identified in 71.4-75% of patients on PFA-100 C-ADP, 83.9-86.8% of patients on VerifyNow P2Y12, and 81.5-83.3% of patients on Multiplate ADP assays. There were no changes in CD62P/CD63 expression (P ≥ 0.18), or consistent changes in leucocyte-platelet complexes in CVD patients overall at 14d or 90d vs. baseline after commencing dipyridamole. Monocyte-platelet complexes increased in the patient subgroup with dipyridamole-HTPR at 14d and 90d on PFA-100, and at 14d on VerifyNow (P ≤ 0.04), but not in those without dipyridamole-HTPR. DISCUSSION: Additional antiplatelet effects of dipyridamole are detectable under high and low shear stress conditions with user-friendly platelet function/reactivity tests ex vivo. Increasing circulating monocyte-platelet complexes over time are associated with dipyridamole-HTPR.


Assuntos
Isquemia Encefálica , Ataque Isquêmico Transitório , AVC Isquêmico , Acidente Vascular Cerebral , Difosfato de Adenosina/metabolismo , Difosfato de Adenosina/farmacologia , Aspirina/farmacologia , Aspirina/uso terapêutico , Plaquetas , Isquemia Encefálica/metabolismo , Dipiridamol/metabolismo , Dipiridamol/farmacologia , Dipiridamol/uso terapêutico , Humanos , Ataque Isquêmico Transitório/tratamento farmacológico , Ativação Plaquetária , Inibidores da Agregação Plaquetária/farmacologia , Inibidores da Agregação Plaquetária/uso terapêutico , Estudos Prospectivos
3.
QJM ; 114(4): 233-237, 2021 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-32298454

RESUMO

BACKGROUND: The use of anti-epileptic drugs (AEDs) in women of childbearing age (WCBA) necessitates careful counselling regarding reproductive-related issues. AIM: (i) To compare documentation of appropriate counselling regarding reproductive-related issues in WCBA prescribed AEDs for non-epilepsy vs. epilepsy indications, and (ii) to examine whether the frequency of counselling improved after introduction of 'standardized typed advice'. DESIGN: Retrospective audit and quality assessment and improvement programme. METHODS: We analysed medical records of all WCBA prescribed gabapentin, pregabalin, topiramate, valproate or carbamazepine by a general neurology clinical service before (Study period A) and after (Study period B) introduction of standardized typed passages regarding potential teratogenicity ± interactions with hormonal contraception at a university teaching hospital. The χ2 test or the Fisher's exact test was employed, as appropriate. RESULTS: In WCBA prescribed AEDs for non-epilepsy indications, documentation of appropriate counselling regarding potential teratogenicity improved from 49% (17/35 patients) in Period A to 79% (27/34 patients) in Period B (P = 0.008). The frequency of counselling regarding teratogenicity was higher in patients prescribed AEDs for epilepsy compared with non-epilepsy indications in Study period A (100% vs. 49%, P = 0.002), but was no longer significantly different in Study period B (86% vs. 79%, P = 0.64). Documentation of counselling regarding potential interaction of enzyme-inducing AEDs with hormonal contraception did not significantly change between study periods. CONCLUSION: Significant improvements in documentation regarding potential teratogenicity of AEDs prescribed for non-epilepsy indications can be achieved by introducing standardized, typed passages copied to patients. Such a practice change is practical and widely applicable to neurological and non-neurological practice worldwide.


Assuntos
Anticonvulsivantes/efeitos adversos , Aconselhamento , Epilepsia/tratamento farmacológico , Complicações na Gravidez/tratamento farmacológico , Adulto , Anticonvulsivantes/uso terapêutico , Anticoncepção/efeitos adversos , Anticoncepcionais Orais Hormonais/uso terapêutico , Feminino , Humanos , Gravidez , Complicações na Gravidez/prevenção & controle , Estudos Retrospectivos
4.
Gastroenterol Res Pract ; 2019: 1592306, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30881445

RESUMO

BACKGROUND: As the malignant potential of sessile serrated lesions/polyps (SSL/Ps) and traditional serrated adenomas (TSAs) has been clearly demonstrated, it is important that serrated polyps are identified and correctly classified histologically. AIM: Our aim was to characterize the clinicopathological features of a series of SSL/Ps & TSAs, to assess the accuracy of the pathological diagnosis, the incidence, and the rate of dysplasia in SSL/Ps & TSAs. METHODS: We identified all colorectal serrated polyps between 01/01/2004 and 31/05/2016, by searching the laboratory information system for all cases assigned a "serrated adenoma" SNOMED code. All available and suitable slides were reviewed by one pathologist, who was blinded to the original diagnosis and the site of the polyp. Subsequently discordant cases, SSL/Ps with dysplasia, and all TSAs were reviewed by a second pathologist. RESULTS: Over a 149-month period, 759 "serrated adenoma" polyps were identified, with 664 (from 523 patients) available for review. 41.1% were reviewed by both pathologists; 15.1% (100/664) were reclassified, with the majority being changed from SSL/P to hyperplastic polyp (HYP) (66/664; 9.9%). 80.3% of these HYPs were located in the left colon, and the majority exhibited prolapse effect. There were 520 SSL/Ps (92.2%) & 40 TSAs (7.1%). The majority of SSL/Ps were in the right colon (86.7%) and were small (64.5% <1 cm), while most TSAs were in the left colon (85.7%) and were large (73.1%≥1 cm). 6.7% of SSL/Ps exhibited dysplasia, the majority of which were large (66.7%≥1 cm). Following consensus review, 13/520 (2.5%) SSL/Ps were downgraded from SSL/P with dysplasia to SSL/P without dysplasia. Detection of SSL/Ps peaked in the most recent years reviewed (87.5% reported between 2013 and 2016, inclusive), coinciding with the introduction of "BowelScreen" (the Irish FIT-based colorectal cancer screening programme). CONCLUSIONS: Awareness of, and adherence to, diagnostic criteria is essential for accurate classification of colorectal polyps.

5.
Sci Rep ; 7: 40424, 2017 01 20.
Artigo em Inglês | MEDLINE | ID: mdl-28106081

RESUMO

How temperate bacteriophages play a role in microbial infection and disease progression is not fully understood. They do this in part by carrying genes that promote positive evolutionary selection for the lysogen. Using Biolog phenotype microarrays and comparative metabolite profiling we demonstrate the impact of the well-characterised Shiga toxin-prophage ϕ24B on its Escherichia coli host MC1061. As a lysogen, the prophage alters the bacterial physiology by increasing the rates of respiration and cell proliferation. This is the first reported study detailing phage-mediated control of the E. coli biotin and fatty acid synthesis that is rate limiting to cell growth. Through ϕ24B conversion the lysogen also gains increased antimicrobial tolerance to chloroxylenol and 8-hydroxyquinoline. Distinct metabolite profiles discriminate between MC1061 and the ϕ24B lysogen in standard culture, and when treated with 2 antimicrobials. This is also the first reported use of metabolite profiling to characterise the physiological impact of lysogeny under antimicrobial pressure. We propose that temperate phages do not need to carry antimicrobial resistance genes to play a significant role in tolerance to antimicrobials.


Assuntos
Antibacterianos/farmacologia , Bacteriófagos/metabolismo , Toxina Shiga/metabolismo , Área Sob a Curva , Proliferação de Células/efeitos dos fármacos , Análise Discriminante , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Resistência a Canamicina/efeitos dos fármacos , Lisogenia/efeitos dos fármacos , Metabolômica , Análise Multivariada , Pressão Osmótica , Oxiquinolina/farmacologia , Xilenos/farmacologia
6.
J Clin Microbiol ; 54(12): 2990-2999, 2016 12.
Artigo em Inglês | MEDLINE | ID: mdl-27707938

RESUMO

Histoplasma capsulatum var. farciminosum, the causative agent of epizootic lymphangitis (EZL), is endemic in parts of Africa. Diagnosis based on clinical signs and microscopy lacks specificity and is a barrier to further understanding this neglected disease. Here, a nested PCR method targeting the internal transcribed spacer (ITS) region of the rRNA operon was validated for application to equine clinical samples. Twenty-nine horses with signs of EZL from different climatic regions of Ethiopia were clinically examined. Blood samples and aspirates of pus from cutaneous nodules were taken, along with blood from a further 20 horses with no cutaneous EZL lesions. Among the 29 horses with suspected cases of EZL, H. capsulatum var. farciminosum was confirmed by extraction of DNA from pus and blood samples from 25 and 17 horses, respectively. Positive PCR results were also obtained with heat-inactivated pus (24 horses) and blood (23 horses) spotted onto Whatman FTA cards. Two positive results were obtained among blood samples from 20 horses that did not exhibit clinical signs of EZL. These are the first reports of the direct detection of H. capsulatum var. farciminosum in equine blood and at high frequency among horses exhibiting cutaneous lesions. The nested PCR outperformed conventional microscopic diagnosis, as characteristic yeast cells could be observed only in 14 pus samples. The presence of H. capsulatum var. farciminosum DNA was confirmed by sequencing the cloned PCR products, and while alignment of the ITS amplicons showed very little sequence variation, there was preliminary single nucleotide polymorphism-based evidence for the existence of two subgroups of H. capsulatum var. farciminosum This molecular diagnostic method now permits investigation of the epidemiology of EZL.


Assuntos
Histoplasma/isolamento & purificação , Histoplasmose/veterinária , Doenças dos Cavalos/diagnóstico , Linfangite/diagnóstico , Linfangite/veterinária , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase/métodos , Animais , Sangue/microbiologia , Corynebacterium pseudotuberculosis/isolamento & purificação , DNA Espaçador Ribossômico/genética , Diagnóstico Diferencial , Etiópia , Histoplasma/classificação , Histoplasmose/diagnóstico , Histoplasmose/microbiologia , Doenças dos Cavalos/microbiologia , Cavalos , Linfangite/microbiologia , Supuração/microbiologia
7.
Reprod Domest Anim ; 48(3): 525-8, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23331295

RESUMO

The bacterial load and degree of antibiotic resistance present in untreated and antibiotic-treated semen samples were investigated in five bulls standing at a cattle-breeding centre. Bacterial load was determined by colony counts from semen samples cultured on brain heart infusion and nutrient agar plates. Antibiotic resistance in these bacteria was assessed by measuring the diameter of bacterial growth inhibition zones around discs containing different concentrations of antibiotics. Representative antibiotic-resistant bacterial isolates were selected for identification. Untreated semen contained few culturable bacteria, and all were completely sensitive to gentamycin, spectinomycin and lincomycin: six of the isolates showed some resistance to tylosin. In semen to which antibiotics had been added as part of the routine production process, two isolates were sensitive to all of the antibiotics tested, and the remainder were resistant to all. Resistant Gram-negative isolates that were identified included Pseudomonas and Stenotrophomonas spp. both in the class Gammaproteobacteria and a Sphingomonas sp. which is in the class Alphaproteobacteria.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Bactérias Gram-Negativas/efeitos dos fármacos , Sêmen/microbiologia , Animais , Bovinos , Masculino
8.
J Dairy Sci ; 95(12): 6949-56, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22999287

RESUMO

Forty samples of raw milk cheese and 25 samples of raw milk itself were subjected to enrichment culture for Shiga-toxigenic Escherichia coli (STEC), and a single Shiga toxin 2- (Stx(2)) positive strain was obtained from one of the cheese samples. Thus, aged cheeses in which the curd is subsequently heat treated (48°C) cannot be presumed to be STEC free. Infective Stx(2) bacteriophages were induced from 3 STEC strains isolated elsewhere from raw milk and 1 STEC strain from aged cheese sourced in Italy. Data on E. coli host range, morphology, genome size, and genetic variation determined by restriction fragment length polymorphism and multi-locus genotyping are presented. Although all 4 bacteriophages were found to be short-tailed Podoviridae, they exhibited considerable variation in both genome size and content. This extended to the Stx(2) genes themselves, whose sequences contained several point mutations, but these did not translate to amino acid substitutions.


Assuntos
Laticínios/microbiologia , Podoviridae/genética , Toxina Shiga II/genética , Escherichia coli Shiga Toxigênica/isolamento & purificação , Animais , Sequência de Bases , Bovinos , Queijo/microbiologia , Queijo/virologia , Laticínios/virologia , Itália , Microscopia Eletrônica de Transmissão , Leite/microbiologia , Leite/virologia , Dados de Sequência Molecular , Escherichia coli Shiga Toxigênica/virologia
9.
J Genet ; 91(2): 199-204, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22942090

RESUMO

The ad-8 gene of Neurospora crassa, in addition to being used for the study of purine biology, has been extensively studied as a model for gene structure, mutagenesis and intralocus recombination. Because of this there is an extensive collection of well-characterized N. crassa ad-8 mutants in the Fungal Genetics Stock Center collection. Among these are spontaneous mutants and mutants induced with X-ray, UV or chemical mutagens. The specific lesions in these mutants have been genetically mapped at high resolution. We have sequenced the ad-8 locus from 13 of these mutants and identified the molecular nature of the mutation in each strain. We compare the historical fine-structure map to the DNA and amino acid sequence of each allele. The placement of the individual lesions in the fine-structure map was more accurate at the 5' end of the gene and no mutants were identified in the 3' untranslated region of this gene. We additionally analysed ad-8(+) alleles in 18 N. crassa strains subjected to whole-genome sequence analysis and describe the variability among Neurospora strains and among fungi and other organisms.


Assuntos
Adenilossuccinato Sintase/genética , Proteínas Fúngicas/genética , Neurospora crassa/genética , Alelos , Análise Mutacional de DNA , Mutagênese , Mutação , Neurospora crassa/enzimologia , Filogenia
10.
Anim Genet ; 42(5): 535-43, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21906104

RESUMO

The sheep has worldwide agricultural importance, yet the genetic control of the immune responses underlying susceptibility or resistance to ovine disease is little understood. Here, we identify six novel polymorphisms in the ovine immune response genes interferon-γ (IFNG), tumour necrosis factor-α (TNF), interleukin-1ß (IL1B) and interleukin-4 (IL4) in pedigree Charollais flocks. We confirm the presence of previously reported polymorphisms in IFNG and IL1B in Charollais. Restriction fragment length polymorphism (RFLP) genotyping assays have been developed for four polymorphisms, IFNGg.168C>T, IFNGg.285A>G, IL1Bg.689C>T and TNFg.3UTRA>G, and a Taqman genotyping assay has been developed for IL4g.485C>T. The previously described IL2g.647C>T polymorphism is adapted for RFLP analysis. Allele frequencies are described in Charollais, Lleyn and Suffolk cross sheep. Polymorphisms are typed in both Charollais ewes and lambs and analysed against abortion phenotypes. A subset of animals have also been analysed for the presence of Toxoplasma gondii, an abortion-causing protozoan. The IFNGg.168T allele is shown to be associated with increased risk of a ewe having an abortion, while the IFNGg.285G allele is associated with increased risk of a lamb being aborted. These assays provide tools for the investigation of the genetic basis of other phenotypes in sheep, including infectious disease susceptibility.


Assuntos
Aborto Animal/genética , Aborto Animal/imunologia , Citocinas/genética , Polimorfismo Genético , Doenças dos Ovinos/genética , Doenças dos Ovinos/imunologia , Animais , Citocinas/imunologia , Feminino , Gravidez , Carneiro Doméstico , Toxoplasma
11.
Ir J Med Sci ; 180(2): 395-9, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20821069

RESUMO

BACKGROUND: Despite understaffing of neurology services in Ireland, the demand for liaison neurologist input into the care of hospital inpatients is increasing. This aspect of the workload of the neurologist is often under recognised. AIMS/METHODS: We prospectively recorded data on referral and service delivery patterns to a liaison neurology service, the neurological conditions encountered, and the impact of neurology input on patient care. RESULTS: Over a 13-month period, 669 consults were audited. Of these, 79% of patients were seen within 48 h and 86% of patients were assessed by a consultant neurologist before discharge. Management was changed in 69% cases, and discharge from hospital expedited in 50%. If adequate resources for neurological assessment had been available, 28% could have been seen as outpatients, with projected savings of 857 bed days. CONCLUSIONS: Investment in neurology services would facilitate early accurate diagnosis, efficient patient and bed management, with substantial savings.


Assuntos
Hospitalização/economia , Doenças do Sistema Nervoso/diagnóstico , Neurologia , Encaminhamento e Consulta/estatística & dados numéricos , Humanos , Irlanda , Tempo de Internação , Doenças do Sistema Nervoso/terapia , Estudos Prospectivos , Fatores de Tempo , Recursos Humanos
12.
Nucleic Acids Res ; 39(6): 2116-29, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21062824

RESUMO

Shigatoxigenic Escherichia coli emerged as new food borne pathogens in the early 1980s, primarily driven by the dispersal of Shiga toxin-encoding lambdoid bacteriophages. At least some of these Stx phages display superinfection phenotypes, which differ significantly from lambda phage itself, driving through in situ recombination further phage evolution, increasing host range and potentially increasing the host's pathogenic profile. Here, increasing levels of Stx phage Φ24(B) integrase expression in multiple lysogen cultures are demonstrated along with apparently negligible repression of integrase expression by the cognate CI repressor. The Φ24(B) int transcription start site and promoter region were identified and found to differ from in silico predictions. The unidirectional activity of this integrase was determined in an in situ, inducible tri-partite reaction. This indicated that Φ24(B) must encode a novel directionality factor that is controlling excision events during prophage induction. This excisionase was subsequently identified and characterized through complementation experiments. In addition, the previous proposal that a putative antirepressor was responsible for the lack of immunity to superinfection through inactivation of CI has been revisited and a new hypothesis involving the role of this protein in promoting efficient induction of the Φ24(B) prophage is proposed.


Assuntos
Colífagos/enzimologia , Colífagos/genética , DNA Nucleotidiltransferases/metabolismo , Integrases/metabolismo , Proteínas Virais/metabolismo , Biologia Computacional , DNA Nucleotidiltransferases/química , Integrases/genética , Modelos Moleculares , Escherichia coli Shiga Toxigênica/virologia , Sítio de Iniciação de Transcrição , Proteínas Virais/química
13.
Appl Environ Microbiol ; 68(4): 2049-53, 2002 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-11916731

RESUMO

Primer sets specific for 16S rRNA genes were designed for four phylogenetic groups of clostridia known to contain mesophilic cellulolytic species. Specific amplification of these groups from landfill leachate DNA extracts demonstrated the widespread occurrence of clostridia from the Clostridium thermocellum and C. leptum groups. In contrast, the C. botulinum group was never detected, and the C. coccoides-C. lentocellum group was only occasionally detected. Amplification products were analyzed by temporal thermal gel electrophoresis to generate profiles of the clostridial groups and to identify dominant bands. Sequence analysis of 17 landfill clones confirmed that the primers were specific for the clostridial subgroups and that the cloned sequences had a close relationship with known cellulose-degrading clostridia. The primers have therefore been authenticated for use in the rapid identification of clostridia in anaerobic environments.


Assuntos
Clostridium/classificação , Clostridium/isolamento & purificação , Primers do DNA/genética , Genes de RNAr/genética , Eliminação de Resíduos , Anaerobiose , Clostridium/genética , Eletroforese em Gel de Ágar/métodos , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
14.
J Appl Microbiol ; 92(1): 127-33, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-11849336

RESUMO

AIMS: To determine whether composting with animal manure can be used to effectively remediate soil from a pentachlorophenol (PCP)-contaminated site, and to establish the fate of the degraded xenobiotic. METHODS AND RESULTS: Contaminated soil from a sawmill site was mixed with farm animal manure and composted in a 0.5 m3 silo under fully aerobic conditions. The disappearance and fate of PCP was monitored by gas chromatography (GC-ECD) and extensive mineralization confirmed in experiments with 14C-radiolabelled PCP. The disappearance of PCP was rapid and virtually complete within 6 days, prior to the onset of thermophilic conditions. Dechlorination of the PCP was found to be both reductive and sequential. CONCLUSIONS: PCP removal from contaminated soil by aerobic composting with animal manure is efficient and proceeds via reductive dechlorination to virtually complete mineralization. This contrasts with other chlorophenol composting regimes in which mineralization is achieved but dechlorination intermediates do not accumulate to detectable levels. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study demonstrate that anaerobic reductive dechlorination can proceed in an aerobic composting environment and contribute to efficient pentachlorophenol removal. Farmyard manure composts may represent a rapid, low-cost, low-technology option for treatment of chlorophenol-contaminated soils.


Assuntos
Agricultura , Bactérias/metabolismo , Esterco/microbiologia , Pentaclorofenol/metabolismo , Poluentes do Solo/metabolismo , Biodegradação Ambiental , Cloro/metabolismo , Oxirredução , Eliminação de Resíduos , Fatores de Tempo
15.
Appl Environ Microbiol ; 67(9): 4335-7, 2001 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-11526041

RESUMO

A verocytotoxigenic bacteriophage isolated from a strain of enterohemorrhagic Escherichia coli O157, into which a kanamycin resistance gene (aph3) had been inserted to inactivate the verocytotoxin gene (vt2), was used to infect Enterobacteriaceae strains. A number of Shigella and E. coli strains were susceptible to lysogenic infection, and a smooth E. coli isolate (O107) was also susceptible to lytic infection. The lysogenized strains included different smooth E. coli serotypes of both human and animal origin, indicating that this bacteriophage has a substantial capacity to disseminate verocytotoxin genes. A novel indirect plaque assay utilizing an E. coli recA441 mutant in which phage-infected cells can enter only the lytic cycle, enabling detection of all infective phage, was developed.


Assuntos
Colífagos/fisiologia , Escherichia coli O157/virologia , Escherichia coli/virologia , Toxina Shiga II/metabolismo , Shigella/virologia , Animais , Bovinos , Colífagos/genética , Colífagos/isolamento & purificação , Humanos , Lisogenia , Toxina Shiga II/genética
16.
Antonie Van Leeuwenhoek ; 79(2): 179-88, 2001 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-11520004

RESUMO

A combination of PCR amplification and oligonucleotide probing was used to investigate the populations of ammonia-oxidisers of the beta-Proteobacteria in the eutrophic and oligotrophic basins of Lake Windermere, a large temperate lake in the English Lake District. Numbers of ammonia-oxidisers (MPN) in the Windermere lakewater were low (< 100 cells ml(-1)) throughout the year with the exception of peaks in August, which coincided with stratification, and November in the South Basin where overturn may have introduced ammonia-oxidising bacteria into the water column. Sediment samples contained larger populations of ammonia oxidisers, usually ca. 10(4) per g. dry weight, which remained relatively constant throughout the seasonal cycle in both Basins. DNA was recovered from lakewater and sediment samples and Nitrosospira and N. europaea-eutropha lineage 16S rRNA genes amplified in a nested PCR reaction, with confirmation of identity by oligonucleotide hybridisation. Nitrosospira 16S rDNA was readily detected in all samples and therefore found to be ubiquitous. In contrast, nitrosomonad DNA of the N. europaea-eutropha lineage could only be detected in the oligotrophic North Basin. Enrichment cultures of lakewater samples only exhibited nitrification at low (0.67 mM) and medium (5 mM) ammonium concentrations, whilst sediment enrichments nitrified at all concentrations tested including high (12.5 mM) ammonium medium. These data suggest that ammonia-oxidiser populations may be physiologically distinguished between lakewater and sediment, and that species distribution in a single lake is non-uniform.


Assuntos
Amônia/metabolismo , Betaproteobacteria/classificação , Betaproteobacteria/crescimento & desenvolvimento , Água Doce/microbiologia , Betaproteobacteria/genética , Betaproteobacteria/isolamento & purificação , Contagem de Colônia Microbiana , Meios de Cultura , DNA Ribossômico/análise , Ecossistema , Eutrofização , Genes de RNAr , Sondas de Oligonucleotídeos , Oxirredução , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/genética
17.
Lett Appl Microbiol ; 30(1): 14-8, 2000 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-10728553

RESUMO

Ammonia-oxidizer numbers decreased under conditions of moisture limitation in litter, fermentation and humus layers of forest soil in the field, but the extent of regrowth after rehydration varied between layers. Nitrosospira 16S rRNA genes were amplified from all layers, regardless of moisture content or soil pH which varied between 4.1 and 5.2. Nitrosomonas spp. were detected less often, but appeared to exhibit more rapid recovery than the Nitrosospira spp. when drought conditions were relieved by rainfall.


Assuntos
Amônia/metabolismo , Bradyrhizobiaceae/isolamento & purificação , Nitrosomonas/isolamento & purificação , Microbiologia do Solo , Bradyrhizobiaceae/crescimento & desenvolvimento , Bradyrhizobiaceae/metabolismo , Contagem de Colônia Microbiana , DNA Bacteriano/análise , DNA Ribossômico/análise , Eletroforese em Gel de Ágar , Nitrosomonas/crescimento & desenvolvimento , Nitrosomonas/metabolismo , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/análise , Solo/análise , Água/análise
18.
Med Pediatr Oncol ; 32(6): 411-5, 1999 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-10358698

RESUMO

BACKGROUND: The treatment of relapsed and refractory leukemia in children remains a challenge. The morbidity of further chemotherapy is considerable, as most patients have already been exposed to intensive multiagent chemotherapy. The FLAG (fludarabine, high-dose cytarabine, and G-CSF) regimen is as intensive but less cardiotoxic because of the avoidance of anthracyclines. PROCEDURE: Nineteen children were treated in two U.K. centers with the FLAG regimen for relapsed and refractory acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL). There were 13 males and 6 females, with an age range of 1.9 to 14.2 years. AML was the diagnosis in 12 children, ALL in 4, biphenotypic leukemia in 3. Eight patients had refractory disease, 11 were in relapse (5 in first relapse, 4 in second, and 2 in third). RESULTS: Complete remission was obtained in 13 patients, partial remission was obtained in 4, and 2 patients were considered nonresponders. There were seven patients alive at 12 months (mean) posttherapy; one of these is awaiting bone marrow transplantation (BMT). All patients experienced grade 4 hematological toxicity; no patient died of infection. Thirteen patients received BMT as consolidation (seven unrelated donor; six sibling allografts). Six of these have died, four due to pneumonitis. CONCLUSIONS: FLAG can be regarded as an effective protocol for inducing remission in a group of heavily pretreated children. Its toxicity is acceptable due to the avoidance of anthracyclines.


Assuntos
Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamento farmacológico , Adolescente , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Transplante de Medula Óssea , Criança , Pré-Escolar , Citarabina/uso terapêutico , Feminino , Fator Estimulador de Colônias de Granulócitos/uso terapêutico , Humanos , Lactente , Leucemia Mieloide Aguda/terapia , Masculino , Leucemia-Linfoma Linfoblástico de Células Precursoras/terapia , Recidiva , Resultado do Tratamento , Vidarabina/análogos & derivados , Vidarabina/uso terapêutico
19.
Bone Marrow Transplant ; 24(12): 1315-22, 1999 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-10627641

RESUMO

Respiratory syncytial virus (RSV) is known to cause acute lung injury in the immunocompromised host, especially recipients of bone marrow allografts. Specific prognostic factors for the development of severe life-threatening disease remain to be identified as does the optimum treatment of established disease. Over a 5-year period the incidence and outcome of RSV in BMT recipients was analysed retrospectively. Prognostic factors assessed included type of transplant, engraftment status at the time of infection, the presence of lower respiratory tract disease, viral genotype and treatment received. During the study period, 26 of 336 (6.3%) allogeneic stem-cell recipients were identified as having RSV. Five patients (19.2%) died as a direct result of RSV. One patient died secondary to an intracranial bleed with concomitant RSV. There were four patients with graft failure (two primary and two secondary) attributable to the presence of RSV, two of whom subsequently died of infections related to prolonged myelosuppression. The presence of lower respiratory tract infection and a poor overall outcome was the only statistically significant association. Unrelated donor transplants and AML as the underlying disease appeared to be associated with a poorer outcome. Engraftment status, viral genotype and RSV treatment received did not correlate with outcome. We conclude that future studies are required to identify early sensitive and reproducible prognostic factors of RSV in the immunocompromised host. The roles of intravenous and nebulised ribavirin need to be clarified by prospective controlled trials.


Assuntos
Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Infecções por Vírus Respiratório Sincicial/etiologia , Adolescente , Adulto , Criança , Pré-Escolar , Genótipo , Sobrevivência de Enxerto , Doença Enxerto-Hospedeiro , Neoplasias Hematológicas/terapia , Humanos , Lactente , Radiografia Torácica , Infecções por Vírus Respiratório Sincicial/tratamento farmacológico , Infecções por Vírus Respiratório Sincicial/economia , Estudos Retrospectivos , Ribavirina/economia , Ribavirina/uso terapêutico , Taxa de Sobrevida , Transplante Homólogo , Resultado do Tratamento
20.
Appl Microbiol Biotechnol ; 46(5-6): 514-20, 1996 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-9008884

RESUMO

Xylanase A (XylA) from Pseudomonas fluorescens subsp. cellulosa consists of an N-terminal non-catalytic cellulose-binding domain joined to a functionally independent C-terminal catalytic domain by a sequence rich in serine residues. Xylanase D (XylD) from Cellulomonas fimi also exhibits a modular structure comprising an N-terminal catalytic domain linked to an internal non-catalytic xylan-binding domain and a C-terminal cellulose-binding domain. To determine the importance of the non-catalytic polysaccharide-binding domains and linker sequences of XylA and XylD in relation to their capacity to hydrolyse pulp xylan and enhance bleachability, purified full-length and modified derivatives of both enzymes were incubated with a hardwood kraft pulp. Deletion of the cellulose-binding domain or linker region from XylA decreased the activity of the enzyme against pulp xylan, but had no significant effect on the capacity of the enzyme to facilitate delignification and reduce pulp kappa number. While full-length and truncated forms of XylD, lacking either the cellulose-binding or the cellulose- and xylan-binding domains, were equally effective in hydrolysing pulp xylan, enzyme derivatives containing a polysaccharide-binding domain were marginally more efficient in reducing pulp kappa number. The reduction in kappa number elicited by full-length and isolated catalytic domains of XylA and XylD was reflected in an increase in the brightness of paper handsheets derived from pretreated pulps. Thus, the polysaccharide-binding domains of XylA and XylD did not appear to confer any advantage in terms of the ability of the enzymes to improve pulp bleachability. However, XylA and XylD, which belong to different glycosyl hydrolase families, differed in their ability to hydrolyse pulp xylan and facilitate the delignification of kraft pulp.


Assuntos
Actinomycetales/enzimologia , Actinomycetales/genética , Celulose/metabolismo , Mutagênese Insercional , Pseudomonas fluorescens/enzimologia , Pseudomonas fluorescens/genética , Xilosidases/genética , Xilosidases/fisiologia , beta-Glucosidase/genética , beta-Glucosidase/fisiologia , Clonagem Molecular , Endo-1,4-beta-Xilanases , Regulação Bacteriana da Expressão Gênica , Papel , Peróxidos/metabolismo , Polissacarídeos Bacterianos/química , Polissacarídeos Bacterianos/fisiologia
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