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1.
Bull Math Biol ; 78(3): 436-67, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26934886

RESUMO

The earliest cell fate decisions in a developing embryo are those associated with establishing the germ layers. The specification of the mesoderm and endoderm is of particular interest as the mesoderm is induced from the endoderm, potentially from an underlying bipotential group of cells, the mesendoderm. Mesendoderm formation has been well studied in an amphibian model frog, Xenopus laevis, and its formation is driven by a gene regulatory network (GRN) induced by maternal factors deposited in the egg. We have recently demonstrated that the axolotl, a urodele amphibian, utilises a different topology in its GRN to specify the mesendoderm. In this paper, we develop spatially structured mathematical models of the GRNs governing mesendoderm formation in a line of cells. We explore several versions of the model of mesendoderm formation in both Xenopus and the axolotl, incorporating the key differences between these two systems. Model simulations are able to reproduce known experimental data, such as Nodal expression domains in Xenopus, and also make predictions about how the positional information derived from maternal factors may be interpreted to drive cell fate decisions. We find that whilst cell-cell signalling plays a minor role in Xenopus, it is crucial for correct patterning domains in axolotl.


Assuntos
Anfíbios/embriologia , Modelos Biológicos , Ambystoma mexicanum/embriologia , Ambystoma mexicanum/genética , Proteínas de Anfíbios/genética , Anfíbios/genética , Animais , Simulação por Computador , Endoderma/embriologia , Regulação da Expressão Gênica no Desenvolvimento , Redes Reguladoras de Genes , Conceitos Matemáticos , Mesoderma/embriologia , Ligantes da Sinalização Nodal/genética , Xenopus laevis/embriologia , Xenopus laevis/genética
2.
Int J Dent Hyg ; 14(3): 209-14, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26198308

RESUMO

OBJECTIVES: Sodium bicarbonate has been shown clinically to be efficacious at removing dental plaque; however, its effect of mechanism against biofilms has not been evaluated in vitro. Here, we used a well-established in vitro plaque biofilm model to investigate the disruption of dental plaque biofilms. METHODS: Biofilms were grown in a constant depth film fermentor for up to 14 days. The fermentor was inoculated with pooled human saliva and growth maintained with artificial saliva. After various time points, replicate biofilms were removed and subjected to treatment at varying concentrations of sodium bicarbonate. Disruption of the plaque was assessed by viable counts and microscopy. RESULTS: The viable count results showed that younger biofilms were less susceptible to the action of sodium bicarbonate; however, biofilms of 7 days and older were increasingly susceptible to the material with the oldest biofilms being the most susceptible. Sixty-seven percentage of sodium bicarbonate slurry was able to reduce the number of organisms present by approx. 3 log10 . These quantitative data were corroborated qualitatively with both confocal and electron microscopy, which both showed substantial qualitative removal of mature biofilms. CONCLUSIONS: The results from this study have shown that sodium bicarbonate is able to disrupt mature dental plaque grown in vitro and that its reported efficacy in maintaining oral hygiene may be related to this key factor.


Assuntos
Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Placa Dentária/tratamento farmacológico , Bicarbonato de Sódio/farmacologia , Bactérias/efeitos dos fármacos , Bactérias/crescimento & desenvolvimento , Reatores Biológicos , Contagem de Colônia Microbiana , Placa Dentária/microbiologia , Microscopia Confocal , Microscopia Eletrônica , Higiene Bucal , Saliva , Saliva Artificial , Bicarbonato de Sódio/administração & dosagem , Fatores de Tempo
3.
J Math Biol ; 67(6-7): 1393-424, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23070212

RESUMO

Nodal signals are key regulators of mesoderm and endoderm development in vertebrate embryos. It has been observed experimentally that in Xenopus embryos the spatial range of Nodal signals is restricted by the signal Antivin (also known as Lefty). Nodal signals can activate both Nodal and Antivin, whereas Antivin is thought to antagonise Nodal by binding either directly to it or to its receptor. In this paper we develop a mathematical model of this signalling network in a line of cells. We consider the heterodimer and receptor-mediated inhibition mechanisms separately and find that, in both cases, the restriction by Antivin to the range of Nodal signals corresponds to wave pinning in the model. Our analysis indicates that, provided Antivin diffuses faster than Nodal, either mechanism can robustly account for the experimental data. We argue that, in the case of Xenopus development, it is wave pinning, rather than Turing-type patterning, that is underlying Nodal-Antivin dynamics. This leads to several experimentally testable predictions, which are discussed. Furthermore, for heterodimer-mediated inhibition to prevent waves of Nodal expression from propagating, the Nodal-Antivin complex must be turned over, and diffusivity of the complex must be negligible. In the absence of molecular mechanisms regulating these, we suggest that Antivin restricts Nodal signals via receptor-mediated, and not heterodimer-mediated, inhibition.


Assuntos
Fatores de Determinação Direita-Esquerda/fisiologia , Mesoderma/embriologia , Modelos Biológicos , Proteína Nodal/fisiologia , Transdução de Sinais/fisiologia , Xenopus/embriologia , Animais , Redes Reguladoras de Genes/fisiologia , Fatores de Determinação Direita-Esquerda/genética , Proteína Nodal/genética , Transdução de Sinais/genética
4.
Bull Math Biol ; 72(6): 1383-407, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20135237

RESUMO

The hormone auxin is implicated in regulating a diverse range of developmental processes in plants. Auxin acts in part by inducing the Aux/IAA genes. The associated pathway comprises multiple negative feedback loops (whereby Aux/IAA proteins can repress Aux/IAA genes) that are disrupted by auxin mediating the turnover of Aux/IAA protein. In this paper, we develop a mathematical model of a single Aux/IAA negative feedback loop in a population of identical cells. The model has a single steady-state. We explore parameter space to uncover a number of dynamical regimes. In particular, we identify the ratio between the Aux/IAA protein and mRNA turnover rates as a key parameter in the model. When this ratio is sufficiently small, the system can evolve to a stable limit cycle, corresponding to an oscillation in Aux/IAA expression levels. Otherwise, the steady-state is either a stable-node or a stable-spiral. These observations may shed light on recent experimental results.


Assuntos
Arabidopsis/crescimento & desenvolvimento , Retroalimentação Fisiológica , Ácidos Indolacéticos/metabolismo , Modelos Biológicos , Reguladores de Crescimento de Plantas/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas
5.
J Theor Biol ; 260(1): 41-55, 2009 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-19490918

RESUMO

In this paper we develop a model of mesendoderm specification in Xenopus laevis based on an existing gene regulation network. The mesendoderm is a population of cells that may contribute to either the mesoderm or endoderm. The model that we develop encompasses the time evolution of transcription factor concentrations in a single cell and is shown to have stable steady states that correspond to mesoderm and anterior mesendodermal cell types, but not endoderm (except in cells where Goosecoid expression is inhibited). Both in vitro and in vivo versions of the model are developed and analysed, the former indicating how cell fate is determined in large part by the concentration of Activin administered to a cell, with the model results comparing favourably with current quantitative experimental data. A numerical investigation of the in vivo model suggests that cell fate is determined largely by a VegT and beta-Catenin pre-pattern, subsequently being reinforced by Nodal. We argue that this sensitivity of the model to a VegT and beta-Catenin pre-pattern indicates that a key VegT self-limiting mechanism (for which there is experimental evidence) is absent from the model. Furthermore, we find that the lack of a steady state corresponding to endoderm is entirely consistent with current in vivo data, and that the in vivo model corresponds to mesendoderm specification on the dorsal, but not the ventral, side of the embryo.


Assuntos
Padronização Corporal/fisiologia , Endoderma/citologia , Mesoderma/citologia , Modelos Biológicos , Xenopus laevis/embriologia , Animais , Padronização Corporal/genética , Diferenciação Celular/genética , Diferenciação Celular/fisiologia , Embrião não Mamífero/fisiologia , Desenvolvimento Embrionário/genética , Desenvolvimento Embrionário/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Redes Reguladoras de Genes/genética , Redes Reguladoras de Genes/fisiologia , Fatores de Transcrição/metabolismo , Xenopus laevis/genética , Xenopus laevis/fisiologia
6.
Lett Appl Microbiol ; 48(4): 472-7, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19228292

RESUMO

AIM: To determine the effect of the surface roughness of denture acrylic on the attachment of Streptococcus oralis. METHODS AND RESULTS: Roughened denture acrylic samples were assessed for bacterial attachment, over time, using microscopy. The area of the image covered by bacteria was calculated and converted into a percentage of the total area sampled. The results showed an increasing bacterial coverage with time of incubation and increasing roughness. Differences were seen between heat cured acrylic and cold cured acrylic. CONCLUSION: This study successfully demonstrated a system for the assessment of the amount of attached bacteria on denture acrylic varying roughness. The system was able to discern the difference in surface area coverage by attached bacteria over a roughness range relevant to brushing dentures with dentifrices. SIGNIFICANCE AND IMPACT OF STUDY: This study provides strong support for the scratches caused by brushing dentures with dentifrice encouraging bacterial attachment. This is likely to have a significant effect on efficacy of denture cleaning, general hygiene and biofilm re-formation between cleaning regimens and may indicate that alternative low abrasive cleaners, such as antimicrobial denture-cleaning tablets, offer a more appropriate regimen.


Assuntos
Resinas Acrílicas/química , Aderência Bacteriana , Dentaduras , Streptococcus oralis/fisiologia , Temperatura Baixa , Contagem de Colônia Microbiana , Dentifrícios , Temperatura Alta , Humanos , Propriedades de Superfície , Escovação Dentária , Cremes Dentais
7.
Lett Appl Microbiol ; 44(3): 262-7, 2007 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-17309502

RESUMO

AIMS: To develop a rapid real-time polymerase chain reaction (PCR) method to detect Gluconobacter and Gluconacetobacter species in electrolyte replacement drinks. METHODS AND RESULTS: Samples of electrolyte replacement drinks were artificially contaminated with Gluconobacter species and then filtered to collect cells. DNA was extracted from the filters and analysed by real-time PCR on the ABI Prism 7000 system, using commercial detection kits for lactic and acetic acid bacteria. In addition, specific primers and Taqman probe were designed and used for the detection of seven Gluconobacter and Gluconacetobacter species. All the assays tested demonstrated a linear range of quantification over four orders of magnitude, suggesting detection levels down to 1 CFU ml(-1) in the original drink. CONCLUSIONS: A real-time PCR method was developed to detect low concentrations of Gluconobacter and Gluconacetobacter sp. in an electrolyte replacement drink. SIGNIFICANCE AND IMPACT OF THE STUDY: Real-time PCR methods allow a rapid, high throughput and automated procedure for the detection of food spoilage organisms. The real-time PCR assay described is as sensitive as the conventional method that involves pre-enrichment, enumeration on a selective agar (typically malt extract agar) and identification with a differential medium (typically Wallerstein nutrient agar). The real-time PCR assay also provides a more rapid rate of detection, with results in less than 24 h following enrichment for Gluconobacter and Gluconacetobacter species.


Assuntos
Bebidas/microbiologia , Gluconacetobacter/isolamento & purificação , Gluconobacter/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Primers do DNA/genética , DNA Bacteriano/genética , DNA Ribossômico/genética , Microbiologia de Alimentos , RNA Ribossômico 16S/genética , Reprodutibilidade dos Testes
8.
Respir Med ; 98(12): 1203-6, 2004 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-15588041

RESUMO

Mycobacterium species adhere to the respiratory mucosa via mucus and fibronectin of extracellular matrix exposed by damaged epithelium. We have investigated whether inhibiting adherence to fibronectin influences subsequent infection of human respiratory tissue by Mycobacterium avium complex and Mycobacterium tuberculosis. Human respiratory tissue was pretreated with mycobacterial fibronectin attachment proteins prior to infection with M. avium complex and M. tuberculosis and the number of recoverable bacteria over time was compared to untreated controls. Inhibition significantly reduced recovery of M. avium complex at 15min (P= 0.02), 7days (P = 0.04), and 14 days (P= 0.03); whereas recovery of M. tuberculosis was only reduced at 15 min (P = 0.01) and not at later timepoints. We conclude that M. avium complex and M. tuberculosis infection of the mucosa proceeds by different mechanisms, since M. tuberculosis infection is independent of fibronectin adherence.


Assuntos
Aderência Bacteriana/fisiologia , Fibronectinas/metabolismo , Complexo Mycobacterium avium/fisiologia , Mycobacterium tuberculosis/fisiologia , Mucosa Respiratória/microbiologia , Adesinas Bacterianas/farmacologia , Aderência Bacteriana/efeitos dos fármacos , Humanos , Complexo Mycobacterium avium/isolamento & purificação , Mycobacterium tuberculosis/isolamento & purificação , Técnicas de Cultura de Órgãos , Mucosa Respiratória/metabolismo
9.
Exp Lung Res ; 30(1): 17-29, 2004.
Artigo em Inglês | MEDLINE | ID: mdl-14967601

RESUMO

Mycobacteria adhere specifically to extracellular matrix (ECM) and mucus with a fibrous, but not globular, appearance, in organ cultures of human respiratory mucosa examined by scanning electron microscopy. Previously, light microscopy sections made of tissue infected for 7 days demonstrated mycobacteria associated with mucus on the organ culture surface, and within submucosal glands in areas of damaged epithelium. The authors have now investigated the interactions between Mycobacterium avium complex (MAC), Mycobacterium tuberculosis (MTB), and Mycobacterium smegmatis (MS) and mucus by preincubating bacteria with purified mucins MUC5AC and MUC5B prior to inoculation onto the organ culture mucosal surface. They have also measured mucin production by the organ culture after mycobacterial infection. Mucus did not cause clumping of mycobacteria. There was a significant (P=.03) increase in the amount of fibrous mucus, but not globular mucus, observed on tissue inoculated with mucins compared to controls. The number of bacteria adhering to ECM was markedly reduced after incubation with mucins, which could indicate a protective effect. Mycobacterial infection did not increase mucin production by the organ culture. Mycobacterial adherence to mucins may play a role in the pathogenicity of mycobacteria in diseases such as cystic fibrosis, bronchiectasis, and chronic obstructive pulmonary disease (COPD), in which there are changes in mucus composition and clearance.


Assuntos
Aderência Bacteriana/fisiologia , Mycobacterium tuberculosis/crescimento & desenvolvimento , Mycobacterium tuberculosis/patogenicidade , Mucosa Nasal/microbiologia , Tuberculose Pulmonar/microbiologia , Ar , Humanos , Microscopia Eletrônica , Microscopia Eletrônica de Varredura , Mucinas/metabolismo , Infecções por Mycobacterium não Tuberculosas/microbiologia , Mycobacterium avium/crescimento & desenvolvimento , Mycobacterium avium/patogenicidade , Mycobacterium smegmatis/crescimento & desenvolvimento , Mycobacterium smegmatis/patogenicidade , Mucosa Nasal/metabolismo , Mucosa Nasal/ultraestrutura , Técnicas de Cultura de Órgãos , Virulência
10.
Respir Med ; 97(4): 375-81, 2003 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-12693797

RESUMO

The pathogenicity of Haemophilus parainfluenzae (Hpi) in the respiratory tract is unclear, in contrast to the accepted pathogenicity of its close relative non-typable H. influenzae. We have investigated the interaction of two Hpi isolates with the mucosa of adenoid and bronchial tissue organ cultures. The adherence of bacteria to the mucosa of organ cultures, the effect of broth culture filtrates on human nasal epithelium, and interleukin (IL)-8 production by A549 cell cultures was investigated. Hpi 4846 adhered infrequently in clusters of pleomorphic cocco-bacilli to areas of epithelial damage, mucus and unciliated cells in adenoid organ culture experiments at 24 h, but not bronchial mucosa. Hpi 3698 was seen in only one adenoid and no bronchial organ cultures at 24 h. In separate experiments, Hpi 3698 was cleared more rapidly from the centre of the adenoid organ culture and was not cultured at 24 h. Although not adhering to the mucosa at 24 h, Hpi 3698, but not Hpi 4846, caused an increase in the amount of epithelial damage in both types of organ culture. Broth culture filtrates of both strains caused immediate slowing of ciliary beat frequency that progressed, and disrupted epithelial integrity. Dialysed culture filtrates of both strains stimulated IL-8 production by A549 cells, with the culture filtrate of Hpi 3698 being most potent. We conclude that two strains of Hpi varied in their adherence to adenoid tissue, and neither adhered to bronchial tissue. These results lead us to speculate that Hpi is only likely to be a pathogen in the lower respiratory tract when impaired airway defences delay bacterial clearance.


Assuntos
Infecções por Haemophilus/microbiologia , Haemophilus/patogenicidade , Mucosa Respiratória/microbiologia , Infecções Respiratórias/microbiologia , Brônquios/microbiologia , Células Cultivadas , Cílios/fisiologia , Humanos , Interleucina-8/metabolismo , Especificidade da Espécie , Escarro/microbiologia
11.
Thorax ; 58(3): 246-51, 2003 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-12612305

RESUMO

BACKGROUND: The pathogenesis of Mycobacterium avium complex and Mycobacterium tuberculosis in the respiratory tract is poorly understood, as are the reasons for their differing virulence. We have previously shown that their initial adherence to the mucosa is identical. METHODS: The interaction of M avium complex, M tuberculosis, and M smegmatis with human respiratory tissue was investigated in an organ culture model with an air interface. Tissue was infected for intervals up to 14 days and assessed by scanning electron microscopy for adherent bacteria or cultured for recoverable bacteria. RESULTS: The mean number of adherent bacteria/mm(2) (and the viable count of macerated tissue, cfu/ml) at 15 minutes, 3 and 24 hours, 7 and 14 days were: M avium complex 168 (153), 209 (136), 289 (344), 193 (313), 14140 (16544); M tuberculosis 30 (37), 39 (23), 48 (53), 1 (760), 76 (2186); M smegmatis 108 (176), 49 (133), 97 (81), 114 (427), 34 (58), (n=6). There was no significant change in morphology between infected and uninfected tissue or tissue infected with the different species over 14 days. The number of M avium complex on the mucosa and recovered from tissue increased over time (p=0.03). M tuberculosis decreased on the surface, but recoverable bacteria increased (p=0.01). M smegmatis numbers on the mucosa and recovered from tissue decreased. Sectioned tissue showed M avium complex and M tuberculosis in submucosal mucus glands and M tuberculosis penetrating epithelial cells in one experiment. CONCLUSIONS: The initial adherence to the mucosa of the three species was similar, but after 14 days they varied in their interaction with the tissue in a manner compatible with their pathogenicity.


Assuntos
Complexo Mycobacterium avium/patogenicidade , Mycobacterium tuberculosis/patogenicidade , Mucosa Respiratória/microbiologia , Aderência Bacteriana , Células Cultivadas , Humanos , Microscopia Eletrônica de Varredura , Conchas Nasais/microbiologia , Virulência
12.
Tuberculosis (Edinb) ; 82(2-3): 69-78, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-12356457

RESUMO

OBJECTIVE: Endobronchial infection is associated with pulmonary tuberculosis in the majority of cases. We have investigated the adherence of Mycobacterium tuberculosis to the human respiratory mucosa. DESIGN: Organ cultures constructed with human tissue were infected with M. tuberculosis in the presence or absence of mycobacterial fibronectin attachment cell surface proteins and examined by scanning electron microscopy. RESULTS: M. tuberculosis adhered mainly to extracellular matrix (ECM) in areas of mucosal damage, but not to ciliated mucosa, intact extruded cells, basement membrane or collagen fibres. Bacteria also adhered to fibrous but not globular mucus and occasionally to healthy unciliated mucosa, open tight junctions and to extruded cells that had degenerated, exposing their contents. There was a significant reduction (p<0.05) in the number of bacteria adhering to ECM after pre-incubation of bacteria with fibronectin and after pre-incubation of the tissue with M. avium fibronectin attachment protein (FAP) and M. bovis antigen 85B protein, in a concentration dependent manner. The combined effect of FAP and antigen 85B protein was significantly greater than either protein alone. Bacterial adherence to fibrous mucus was not influenced by fibronectin. CONCLUSION: We conclude that M. tuberculosis adheres to ECM in areas of mucosal damage at least in part via FAP and antigen 85B protein.


Assuntos
Aciltransferases , Antígenos de Bactérias , Aderência Bacteriana , Mycobacterium tuberculosis/fisiologia , Mucosa Respiratória/microbiologia , Adesinas Bacterianas/farmacologia , Aderência Bacteriana/efeitos dos fármacos , Proteínas de Bactérias/farmacologia , Brônquios/efeitos dos fármacos , Brônquios/microbiologia , Matriz Extracelular/efeitos dos fármacos , Matriz Extracelular/microbiologia , Matriz Extracelular/patologia , Fibronectinas/farmacologia , Humanos , Microscopia Eletrônica de Varredura , Mycobacterium tuberculosis/efeitos dos fármacos , Mucosa Nasal/efeitos dos fármacos , Mucosa Nasal/microbiologia , Técnicas de Cultura de Órgãos , Mucosa Respiratória/efeitos dos fármacos
13.
Thorax ; 57(6): 477-82, 2002 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12037221

RESUMO

BACKGROUND: The re-emergence of tuberculosis as a global health problem over the past two decades, accompanied by an increase in tuberculosis drug resistance, prompted the development of a comprehensive national surveillance system for tuberculosis drug resistance in 1993. METHODS: The UK Mycobacterial Resistance Network (Mycobnet), which includes all mycobacterial reference and regional laboratories in the UK, collects a minimum dataset on all individuals from whom an initial isolate of Mycobacterium tuberculosis complex has been isolated and submitted by source hospital laboratories. Data sought include susceptibility to first line antibiotics, demographic, geographical, and risk factor information. RESULTS: There were 25 217 reports of initial isolates of M tuberculosis complex in the UK between 1993 and 1999. All were tested for sensitivity to isoniazid, rifampicin, and ethambutol and 12 692 of the isolates were also tested for sensitivity to pyrazinamide and streptomycin. A total of 1523 (6.1%) isolates were resistant to one or more drugs, 1397 isolates (5.6%) were resistant to isoniazid with or without resistance to other drugs, and 299 (1.2%) were multidrug resistant. Although the numbers of drug resistant isolates increased over the period, the proportions remained little changed. Certain groups of people were at a higher risk of acquiring drug resistant tuberculosis including younger men, residents of London, foreign born subjects, patients with a previous history of tuberculosis and those infected with HIV. CONCLUSION: Although the proportion of drug resistant tuberculosis cases appears to be stable in the UK at present, more than one in 20 patients has drug resistant disease at diagnosis and more than one in 100 has multidrug resistant disease. Tuberculosis control measures should be strengthened to minimise the emergence of drug resistance through rapid diagnosis, rapid identification of drug resistance, supervised treatment, and maintenance of comprehensive surveillance.


Assuntos
Tuberculose Resistente a Múltiplos Medicamentos/epidemiologia , Tuberculose Pulmonar/epidemiologia , Infecções Oportunistas Relacionadas com a AIDS/complicações , Infecções Oportunistas Relacionadas com a AIDS/epidemiologia , Adolescente , Adulto , Distribuição por Idade , Idoso , Antituberculosos/uso terapêutico , Distribuição de Qui-Quadrado , Farmacorresistência Bacteriana , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Recidiva , Características de Residência , Fatores de Risco , Distribuição por Sexo , Tuberculose Resistente a Múltiplos Medicamentos/complicações , Tuberculose Resistente a Múltiplos Medicamentos/tratamento farmacológico , Tuberculose Pulmonar/complicações , Tuberculose Pulmonar/tratamento farmacológico , Reino Unido/epidemiologia
14.
Mol Microbiol ; 38(2): 381-91, 2000 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-11069663

RESUMO

Mycobacterium avium complex (MAC) are opportunistic respiratory pathogens that infect non-immunocompromised patients with established lung disease, although they can also cause primary infections. The ability to bind fibronectin is conserved among many mycobacterial species. We have investigated the adherence of a sputum isolate of MAC to the mucosa of organ cultures constructed with human tissue and the contribution of M. avium fibronectin attachment protein (FAP) to the process. MAC adhered to fibrous, but not globular mucus, and to extracellular matrix (ECM) in areas of epithelial damage, but not to intact extruded cells and collagen fibres. Bacteria occasionally adhered to healthy unciliated epithelium and to cells that had degenerated exposing their contents, but never to ciliated cells. The results obtained with different respiratory tissues were similar. Two ATCC strains of MAC gave similar results. There was a significant reduction (P < 0.05) in the number of bacteria adhering to ECM after preincubation of bacteria with fibronectin and after preincubation of the tissue with M. avium FAP in a concentration-dependant manner. The number of bacteria adhering to fibrous mucus was unchanged. Immunogold labelling demonstrated fibronectin in ECM as well as in other areas of epithelial damage, but only ECM bound FAP. A Mycobacterium smegmatis strain had the same pattern of adherence to the mucosa as MAC. When the FAP gene was deleted, the strain demonstrated reduced adherence to ECM, and adherence was restored when the strain was transfected with an M. avium FAP expression construct. We conclude that MAC adheres to ECM in areas of epithelial damage via FAP and to mucus with a fibrous appearance via another adhesin. Epithelial damage exposing ECM and poor mucus clearance will predispose to MAC airway infection.


Assuntos
Adesinas Bacterianas/fisiologia , Complexo Mycobacterium avium , Mucosa Respiratória/microbiologia , Tonsila Faríngea/microbiologia , Tonsila Faríngea/patologia , Tonsila Faríngea/ultraestrutura , Adesinas Bacterianas/metabolismo , Brônquios/microbiologia , Brônquios/patologia , Brônquios/ultraestrutura , Fibronectinas/metabolismo , Humanos , Imuno-Histoquímica , Técnicas de Cultura de Órgãos , Mucosa Respiratória/patologia , Mucosa Respiratória/ultraestrutura , Soluções , Conchas Nasais/microbiologia , Conchas Nasais/patologia , Conchas Nasais/ultraestrutura
15.
J Hosp Infect ; 45(4): 278-82, 2000 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10973744

RESUMO

The efficacy of Sterilox, a super-oxidized water holding a reduction/oxidation potential of greater than 950 mV was compared with the efficacy of glutaraldehyde against clinical isolates of Mycobacterium tuberculosis and Mycobacterium avium-intracellulare. An in use method using an automated bronchoscope washing machine demonstrated that over five cycles, Sterilox with a contact time of 5 min gave log10 reduction factors for M. tuberculosis and M. avium-intracellulare of >6 and >5, respectively. Glutaraldehyde with a contact time of 10 min gave log10 reduction factors for both M. tuberculosis and M. avium-intracellulare of >4, and at a contact time of 20 min >5 each. The non-toxic nature of Sterilox, together with the reduction in viable counts demonstrated in this study, suggest that the solution is an effective alternative mycobactericidal agent to the established disinfectants for the disinfection of bronchoscopes and, therefore, justifies further investigation.


Assuntos
Broncoscópios/microbiologia , Infecção Hospitalar/prevenção & controle , Desinfetantes/farmacologia , Contaminação de Equipamentos/prevenção & controle , Glutaral/farmacologia , Peróxido de Hidrogênio , Complexo Mycobacterium avium/efeitos dos fármacos , Mycobacterium tuberculosis/efeitos dos fármacos , Oxidantes/farmacologia , Humanos , Escarro/microbiologia
16.
J Hosp Infect ; 37(2): 137-43, 1997 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9364262

RESUMO

The efficacy of 0.35% stabilized buffered peracetic acid solution ('Nu-Cidex') against clinical isolates of Mycobacterium tuberculosis, Mycobacterium avium-intracellulare and Mycobacterium chelonae in homogenized sputum was tested. An in-use method, using an automated bronchoscope washing machine, showed that over 10 cycles, at a disinfectant contact time of 5 min, M. tuberculosis and M. chelonae were effectively eradicated from the bronchoscope, even in the absence of detergent and pre-cleaning. M. avium-intracellulare was not eradicated in only one of 10 cycles with contact times of 5 and 10 min, but numbers were reduced by > 7 log10 and > 5 log10, respectively. With detergent present, M. avium-intracellulare was successfully eradicated in all cycles at a contact time of 5 min or greater. The results demonstrate that peracetic acid is an effective mycobactericidal agent for use in the disinfection of bronchoscopes.


Assuntos
Broncoscópios , Desinfetantes/farmacologia , Desinfecção/métodos , Complexo Mycobacterium avium/efeitos dos fármacos , Mycobacterium chelonae/efeitos dos fármacos , Mycobacterium tuberculosis/efeitos dos fármacos , Ácido Peracético/farmacologia , Contaminação de Equipamentos , Humanos , Complexo Mycobacterium avium/isolamento & purificação , Mycobacterium chelonae/isolamento & purificação , Mycobacterium tuberculosis/isolamento & purificação , Escarro/microbiologia
17.
Eur J Clin Microbiol Infect Dis ; 16(2): 166-70, 1997 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-9105848

RESUMO

The rate of recovery and time to detection of mycobacteria from clinical specimens were analysed for specimens inoculated into both radiometric Middlebrook 7H12 (Bactec 12B, originally 12A) medium and Lowenstein-Jensen egg-based medium (with and without sodium pyruvate) over the 15-year period from 1980 to 1994. A total of 19,679 Bactec vials were inoculated together with Lowenstein-Jensen slopes, with 2198 mycobacterial isolates detected. The mean times to detection for Bactec and Lowenstein-Jensen were 11.7 days and 23.9 days, respectively. In 195 cases mycobacteria were isolated from Bactec and not from Lowenstein-Jensen, whereas the reverse was true in 42 cases. The number of contaminated Bactec vials was 488 and the number of contaminated Lowenstein-Jensen slopes 448.


Assuntos
Técnicas de Laboratório Clínico/métodos , Infecções por Mycobacterium/diagnóstico , Mycobacterium/crescimento & desenvolvimento , Técnicas Bacteriológicas , Meios de Cultura/metabolismo , Humanos , Sensibilidade e Especificidade
18.
J Clin Pathol ; 38(10): 1179-84, 1985 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-4056070

RESUMO

To determine the functional importance of the low B12 and red cell folate concentrations repeatedly observed in the elderly 200 consecutive patients admitted to a geriatric unit were studied. Forty six of the patients had low serum concentrations of B12 (15), red cell folate (26), or both (five). Serum B12 and red cell folate concentrations correlated with mean cell volume, and serum B12 correlated with the neutrophil lobe count. Bone marrow deoxyuridine suppression was abnormal in 35% of the patients with low vitamin concentrations, but 55% of those with abnormal deoxyuridine suppression had morphologically normal bone marrow, and 73% had a normal mean cell volume. In patients with low vitamin values the deoxyuridine suppressed value correlated with the haemoglobin concentration and neutrophil lobe count. Thus synthesis of thymidylate was impaired by vitamin B12 or folate deficiency in at least 8% of newly admitted elderly patients, many of whom had normal blood counts despite the biochemical disturbance affecting haemopoiesis. A nutritionally depleted diet may have been responsible for many of the low vitamin values.


Assuntos
Eritrócitos/metabolismo , Ácido Fólico/sangue , Vitamina B 12/sangue , Idoso , Medula Óssea/metabolismo , Desoxiuridina , Deficiência de Ácido Fólico/epidemiologia , Hemoglobinas/análise , Humanos , Contagem de Leucócitos , Neutrófilos , Albumina Sérica/metabolismo , Tiroxina/sangue , Deficiência de Vitamina B 12/epidemiologia
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