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OBJECTIVES: To test whether providing psychological self-help materials would significantly lower the incidence of post-traumatic stress disorder (PTSD) at 6-12 weeks postnatally. DESIGN: Open-label randomised controlled trial, with blinded outcome assessment. SETTING: Community midwifery services in two National Health Service (NHS) trusts in the North West. SAMPLE: A cohort of 2419 women receiving normal NHS postnatal care. METHODS: Midwives screened women for traumatic birth experience; 678 women who screened positively (28.1%) were randomly allocated to self-help with usual care (n = 336) or to usual care alone (n = 342). The self-help materials were a leaflet and online film designed to prevent the development of PTSD after trauma exposure through explaining how to manage early psychological responses. MAIN OUTCOME MEASURE: The primary outcome was a composite of diagnostic and subdiagnostic PTSD at 6-12 weeks postnatally using the gold-standard Clinician-Administered PTSD Scale (CAPS-5) interview. RESULTS: Of the 678 women correctly randomised plus the nine women randomised in error, 478 (70.5%) were followed up. Diagnostic or subdiagnostic PTSD rates at follow-up did not differ between groups who received self-help (26.7%, 65/243) or usual care alone (26.2%, 64/244) (intention-to-treat analysis: RR 1.02, 95% CI 0.68-1.53). Findings remained consistent in the per-protocol analysis (RR 1.04, 95% CI 0.85-1.27). Women viewed the materials very positively. There were no adverse effects. Health economic micro-costing indicated implementation would be very low cost. CONCLUSIONS: Many women experience a traumatic birth and risk developing PTSD, but self-help strategies without professional support are insufficient and should not be routinely introduced. TWEETABLE ABSTRACT: Self-help information alone does not reduce the number of women developing PTSD after a traumatic childbirth.
Assuntos
Intervenção Baseada em Internet , Complicações do Trabalho de Parto , Folhetos , Parto/psicologia , Qualidade de Vida , Transtornos de Estresse Pós-Traumáticos , Adulto , Erros de Diagnóstico/prevenção & controle , Feminino , Humanos , Tocologia/métodos , Complicações do Trabalho de Parto/diagnóstico , Complicações do Trabalho de Parto/prevenção & controle , Complicações do Trabalho de Parto/psicologia , Gravidez , Técnicas Psicológicas , Autogestão/métodos , Transtornos de Estresse Pós-Traumáticos/diagnóstico , Transtornos de Estresse Pós-Traumáticos/etiologia , Transtornos de Estresse Pós-Traumáticos/prevenção & controle , Transtornos de Estresse Pós-Traumáticos/psicologia , Resultado do TratamentoRESUMO
HLA-G is a non-classical class I HLA antigen, normally expressed in high levels only on extravillous cytotrophoblast. It has immunosuppressive properties in pregnancy and has also been found to be upregulated on leucocytes in viral infection. In this study, proportions of all leucocyte subsets expressing HLA-G were found to be low in healthy subjects positive or negative for cytomegalovirus (CMV). Significantly greater proportions of CD4+ CD69+ and CD56+ T cells expressed HLA-G compared to other T cells. However, following stimulation with CMV antigens or intact CMV, proportions of CD4+, CD8+, CD69+ and CD56+ T cells, and also B cells expressing HLA-G, were significantly increased in CMV+ subjects. Despite some subjects having alleles of HLA-G associated with high levels of expression, no relationship was found between HLA-G genotype and expression levels. Purified B cells from CMV+ subjects stimulated in mixed culture with CMV antigens showed significantly increased HLA-G mRNA expression by real-time polymerase chain reaction. Serum levels of soluble HLA-G were similar in CMV- and CMV+ subjects but levels in culture supernatants were significantly higher in cells from CMV+ than from CMV- subjects stimulated with CMV antigens. The HLA-G ligand KIR2DL4 was mainly expressed on NK cells and CD56+ T cells with no differences between CMV+ and CMV- subjects. Following stimulation with IL-2, an increase in the proportion of CD56+ T cells positive for KIR2DL4 was found, together with a significant decrease in CD56dimCD16+ NK cells. The results show that CMV influences HLA-G expression in healthy subjects and may contribute to viral immune evasion.
Assuntos
Citomegalovirus/imunologia , Antígenos HLA-G/metabolismo , Leucócitos/imunologia , Leucócitos/virologia , Receptores KIR2DL4/metabolismo , Adulto , Anticorpos Antivirais/sangue , Antígenos Virais/administração & dosagem , Proliferação de Células , Citomegalovirus/patogenicidade , Infecções por Citomegalovirus/genética , Infecções por Citomegalovirus/imunologia , Feminino , Antígenos HLA-G/genética , Humanos , Evasão da Resposta Imune , Técnicas In Vitro , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/virologia , Leucócitos/classificação , Ligantes , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores KIR2DL4/genética , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/virologia , Adulto JovemRESUMO
BACKGROUND: Aspiration lung disease (ALD) is a common cause of respiratory morbidity in children and adults with severe neurodisability (sND). Recent studies suggest that chronic microaspiration of gastric contents is associated with mild rather than low, airway acidification. We investigated inflammatory responses to infection by airway epithelial cells (AECs) exposed to weakly acidic media. METHODS: Using pH measurements from children with sND at high risk of ALD as a guide, we incubated AECs in weakly acidic (pH5.5-7.4) media alone; in combination with lipopolysaccharide (LPS); or prior to LPS stimulation at normal pH. Interleukin (IL) -6 and IL-8 expression were measured. RESULTS: IL-6/8 expression in AECs simultaneously exposed to weakly acidic media and LPS for 4 h was reduced with no effect on cell viability. Pre-incubation of AECs at weakly acidic pH also reduced subsequent LPS-induced cytokine expression. Suppression of inflammation was greatest at lower pHs (pH 5.5-6.0) for prolonged periods (16/24 h), but this also adversely affected cell viability. CONCLUSION: AEC inflammatory responses to bacterial stimuli is markedly reduced in a mildly acidic environment.
Assuntos
Doenças do Sistema Nervoso Central/complicações , Células Epiteliais/metabolismo , Mediadores da Inflamação/metabolismo , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Pulmão/metabolismo , Aspiração Respiratória de Conteúdos Gástricos/etiologia , Linhagem Celular , Sobrevivência Celular , Doenças do Sistema Nervoso Central/imunologia , Doenças do Sistema Nervoso Central/metabolismo , Regulação para Baixo , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/imunologia , Humanos , Concentração de Íons de Hidrogênio , Mediadores da Inflamação/imunologia , Interleucina-6/genética , Interleucina-6/imunologia , Interleucina-8/genética , Interleucina-8/imunologia , Lipopolissacarídeos/farmacologia , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Aspiração Respiratória de Conteúdos Gástricos/imunologia , Aspiração Respiratória de Conteúdos Gástricos/fisiopatologia , Fatores de TempoRESUMO
We examined lack of private vehicle access and 30 minutes or longer public transportation travel time to mammography facilities for women 40 years of age or older in the urban areas of Boston, Philadelphia, San Antonio, San Diego, Denver, and Seattle to identify transit marginalized populations - women for whom these travel characteristics may jointly present a barrier to clinic access. This ecological study used sex and race/ethnicity data from the 2010 US Census and household vehicle availability data from the American Community Survey 2008-2012, all at Census tract level. Using the public transportation option on Google Trip Planner we obtained the travel time from the centroid of each census tract to all local mammography facilities to determine the nearest mammography facility in each urban area. Median travel times by public transportation to the nearest facility for women with no household access to a private vehicle were obtained by ranking travel time by population group across all U.S. census tracts in each urban area and across the entire study area. The overall median travel times for each urban area for women without household access to a private vehicle ranged from a low of 15 minutes in Boston and Philadelphia to 27 minutes in San Diego. The numbers and percentages of transit marginalized women were then calculated for all urban areas by population group. While black women were less likely to have private vehicle access, and both Hispanic and black women were more likely to be transit marginalized, this outcome varied by urban area. White women constituted the largest number of transit marginalized. Our results indicate that mammography facilities are favorably located for the large majority of women, although there are still substantial numbers for whom travel may likely present a barrier to mammography facility access.
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Traditional interlocked intramedullary (IM) nails have recently been modified to provide enhanced angular stability. These so-called 'angle-stable' IM nails are designed to eliminate construct toggle and also provide increased axial, bending, and torsional stiffness. While this added stability is needed for small fracture gaps to heal, angle-stable nails may be too stiff for large fracture gaps to unite. Even though relative stability is recommended for large fracture gaps, recent in vivo data indicates that traditional nails may allow for too much motion for healing to occur. The current study evaluated a modified technique for implanting an angle-stable nail which allows for an intermediate amount of stability. The compliance of the nail construct was adjusted by over-drilling the near cortex interlocking hole. This led to increased construct motion in torsion, but less so in axial compression and bending. This modification creates stability which is partway between angle-stable and traditional IM nail designs. These findings were unchanged after 50,000 fatigue loading cycles. By carefully selecting the magnitude of over-drilling, the compliance of the construct can easily be modified as it is being implanted. This design modification may lead to more reliable fracture union since the surgeon can tailor the nail compliance to the injury and bone quality.
Assuntos
Pinos Ortopédicos , Fixação Intramedular de Fraturas/métodos , Fraturas Ósseas/cirurgia , Fixadores Internos , Fenômenos Biomecânicos , Humanos , Desenho de Prótese , Suporte de CargaRESUMO
BACKGROUND: The mechanisms regulating antibody expression within the human lung during airway infection are largely unknown. In this study, our objectives were to determine if infection with respiratory syncytial virus (RSV) upregulates expression of the B cell differentiation factors A proliferation inducing ligand (APRIL) and B cell activating factor of the TNF family (BAFF), if this is a common feature of viral airway infection, and how this is regulated in human airway epithelial cells. METHODS: We measured BAFF and APRIL protein expression in bronchoalveolar lavage (BAL) fluid from infants with severe RSV disease, and healthy control children, and in nasopharyngeal aspirates from preschool children with other single respiratory viral infections. We also measured mRNA expression in bronchial brushings from RSV-infected infants, and in RSV-infected paediatric primary airway epithelial cell cultures (pAEC). Beas-2B cell cultures were used to examine mechanisms regulating BAFF expression. RESULTS: BAFF protein and mRNA were elevated (in marked contrast with APRIL) in BAL and bronchial brushings, respectively, from RSV-infected infants. BAFF protein was also found in upper airway secretions from children with human metapneumovirus, H1N1, bocavirus, rhinovirus, RSV and Mycoplasma pneumoniae infection. BAFF mRNA and protein were expressed following in vitro RSV infection of both pAEC and Beas-2B cultures, with mRNA expression peaking 12-h postinfection. BAFF induction was blocked by addition of a neutralising anti-interferon-ß antibody or palivizumab. CONCLUSIONS: BAFF, produced through an interferon-ß-dependent process, is a consistent feature of airway infection, and suggests a role for the airway epithelia in supporting protective antibody and B cell responses in the lung.
Assuntos
Fator Ativador de Células B/genética , Bronquiolite/virologia , Infecções por Vírus Respiratório Sincicial/imunologia , Vírus Sinciciais Respiratórios/imunologia , Bronquiolite/fisiopatologia , Lavagem Broncoalveolar , Estudos de Casos e Controles , Células Cultivadas , Criança , Células Epiteliais/metabolismo , Células Epiteliais/virologia , Feminino , Regulação da Expressão Gênica , Humanos , Técnicas In Vitro , Lactente , Recém-Nascido , Interferon gama/genética , Interferon gama/metabolismo , Masculino , RNA Mensageiro/metabolismo , Infecções por Vírus Respiratório Sincicial/diagnóstico , Vírus Sinciciais Respiratórios/metabolismo , Sensibilidade e Especificidade , Índice de Gravidade de Doença , Membro 13 da Superfamília de Ligantes de Fatores de Necrose Tumoral/genética , Membro 13 da Superfamília de Ligantes de Fatores de Necrose Tumoral/metabolismo , Regulação para CimaRESUMO
Controversy exists on the role of leucocytospermia on fertilisation rates and IVF outcomes. The aim of our study was to identify the effect of leucocytes and leucocyte subpopulations on fertilisation rates in an IVF cycle. A prospective comparative study of the leucocyte subpopulations of seminal fluid of partners of women attending an IVF cycle was conducted. The samples underwent immunocytochemical staining. The monoclonal antibodies used in this study include CD3, CD4, CD8 (T Cells), CD14 (monocytes/macrophages), CD16 (granulocytes), CD20 (B Cells), CD45 (Pan Leucocytes), CD56 (natural killer cells) and CD69 (activated T and B Cells). Of 21 patients who were recruited into the study, seven were identified as poor fertilisers (<35%) and 14 were identified as good fertilisers (>60%). Data were analysed with SPSS version 14. The total leucocyte counts (CD45) between the poor and good fertilisers were not statistically significant. The macrophages and the monocytes (CD14) were significantly elevated in the good fertilisers group in comparison with the poor fertilisers (P < 0.05). We also found that T cells (CD2, CD4, CD8) and CD14 (macrophages) correlated significantly (r = 0.47, P value < 0.01) with the fertilisation rate. Our study confirms that the presence of leucocytes does not adversely affect the fertilisation rates and the outcome of an IVF cycle. However, macrophages and the monocytes (CD14) were significantly elevated in the good fertilisers group. The increased phagocytic activity in these individuals might increase their fertilising potential by removing spermatozoa with abnormal morphology.
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Fertilização in vitro , Leucócitos/imunologia , Sêmen/citologia , Sêmen/imunologia , Adulto , Anticorpos Monoclonais , Antígenos CD/metabolismo , Subpopulações de Linfócitos B/imunologia , Feminino , Humanos , Contagem de Leucócitos , Leucócitos/classificação , Leucócitos/citologia , Macrófagos/imunologia , Masculino , Pessoa de Meia-Idade , Monócitos/imunologia , Gravidez , Estudos Prospectivos , Subpopulações de Linfócitos T/imunologia , Resultado do TratamentoRESUMO
BACKGROUND: Respiratory syncytial virus (RSV) infection of airway epithelial cells (AECs) is an important initial event in RSV bronchiolitis. AEC immunological responses are thought to be critical in driving the subsequent inflammation in the airway. This study examined viral replication, cytotoxicity and cytokine production in cultures of primary AECs from children compared with responses to RSV infection in an immortalised epithelial cell line and to those from infants with RSV bronchiolitis. METHODS: RSV replication, proinflammatory cytokine responses and cytotoxicity in RSV-infected primary AEC cultures derived from bronchial brushings from the lungs of children were compared with those seen in BEAS-2B cultures, as well as AECs and bronchoalveolar lavage fluid collected from children with and without RSV bronchiolitis. RESULTS: Viral replication, cytotoxicity and inflammatory cytokine production were greater in primary AEC cultures than in BEAS-2B cells. Different response patterns were observed, with RSV infection of primary AEC cultures causing distinct peaks of viral replication and matched cytotoxic responses. Some primary AEC culture immunological responses, such as interleukin 8, were similar in magnitude to those seen in clinical samples from the lungs of children with RSV bronchiolitis. Although variable amounts of RSV were detected by PCR in freshly isolated primary AECs, RSV was not detected by immunocytochemistry. CONCLUSION: This is one of the first studies to examine comprehensively the responses to RSV infection in primary AEC cultures from children and shows marked differences from those of a commercially available immortalised human cell line but reassuring similarities to results found in vivo. This suggests that future work investigating responses of AECs to RSV infection should use primary AEC cultures.
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Brônquios/patologia , Mucosa Respiratória/patologia , Infecções por Vírus Respiratório Sincicial/patologia , Vírus Sinciciais Respiratórios/fisiologia , Anticorpos Antivirais/análise , Brônquios/virologia , Líquido da Lavagem Broncoalveolar/virologia , Linhagem Celular , Criança , Pré-Escolar , Citocinas/biossíntese , Feminino , Humanos , Imuno-Histoquímica , Lactente , Recém-Nascido , Masculino , Prognóstico , Mucosa Respiratória/metabolismo , Mucosa Respiratória/virologia , Infecções por Vírus Respiratório Sincicial/imunologia , Infecções por Vírus Respiratório Sincicial/virologia , Vírus Sinciciais Respiratórios/isolamento & purificação , Replicação ViralRESUMO
The role of leucocytospermia in male subfertility is a much debated topic despite being a frequent finding. This study aimed to identify the role of leucocytes, leucocyte subpopulations and natural killer cells in male subfertility. Seventy-sex subfertile men attending a regional andrology unit were recruited into this prospective study and subdivided into groups based on their semen analysis. The different leucocyte subpopulations were identified using immunocytochemical staining. Significant levels of CD3 helper T lymphocytes (P < 0.001) were present in the oligospermic, asthenospermic, oligoasthenospermic and obstructive azoospermic group compared to the normospermic group. Significant levels of B cells (P < 0.05) were present in the asthenospermic, oligoasthenospermic and obstructive azoospermic group. The natural killer cells (CD56) were significantly raised in the oligoasthenospermic and obstructive azoospermic group (P < 0.05). Our study suggests that leucocytospermia impairs sperm function through enhanced T helper cell modulation, increased B cell population which leads to increased levels of antisperm antibody and natural killer cells mediated sperm damage. The site of seminal leucocyte production is not necessarily confined to the vas or the epididymis.
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Leucócitos/citologia , Sêmen/citologia , Humanos , Imuno-Histoquímica , Masculino , Estudos ProspectivosRESUMO
Airway complications are a leading cause of morbidity and mortality in anaesthesia. Effective management of a difficult airway requires the timely availability of suitable airway equipment. The Australian and New Zealand College of Anaesthetists has recently developed guidelines for the minimum set of equipment needed for the effective management of an unexpected difficult airway (TG4 [2010] www.anzca.edu.au/resources/professionaldocuments). TG4 [2010] is based on expert consensus, underpinned by wide consultation and an extensive review of the available evidence, which is summarised in a Background Paper (TG4 BP [2010] www.anzca.edu.au/ resources/professional-documents). TG4 [2010] will be reviewed at the end of one year and thereafter every five years or more frequently if necessary. The current paper is reproduced directly from the Background Paper (TG4 BP [2010]).
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Obstrução das Vias Respiratórias/terapia , Anestesia , Intubação Intratraqueal/instrumentação , Respiração Artificial/instrumentação , Respiração Artificial/métodos , Adulto , Criança , Humanos , Máscaras Laríngeas , LaringoscópiosRESUMO
A 51-year-old female patient with Guillain-Barré syndrome was given three times the intended dose of intravenous human immunoglobulin while admitted to a tertiary intensive care unit. The error went unnoticed for seven hours and appears to have been the result of several successive breakdowns in communication between key staff The patient, fortunately, made a full recovery. This report analyses the communication failure and explores possible ways of avoiding similar occurrences in the future.
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Comunicação , Síndrome de Guillain-Barré/tratamento farmacológico , Imunoglobulinas/administração & dosagem , Prontuários Médicos , Erros de Medicação , Overdose de Drogas , Feminino , Humanos , Unidades de Terapia Intensiva , Erros de Medicação/prevenção & controle , Pessoa de Meia-Idade , Resultado do TratamentoRESUMO
BACKGROUND: In respiratory syncytial virus (RSV) bronchiolitis, neutrophils account for >80% of cells recovered from the airways in bronchoalveolar lavage (BAL) fluid. This study investigated neutrophil activation and Toll-like receptor (TLR) expression in the blood and lungs of infants with severe RSV bronchiolitis. METHODS: BAL fluid and (blood) samples were collected from 24 (16) preterm and 23 (15) term infants ventilated with RSV bronchiolitis, and 12 (8) control infants. Protein levels and mRNA expression of CD11b, myeloperoxidase (MPO) and TLRs 2, 4, 7, 8 and 9 were measured in neutrophils. RESULTS: Blood neutrophils had more CD11b in preterm and term infants with RSV bronchiolitis than control infants (p<0.025) but similar amounts of MPO. BAL fluid neutrophils from infants with RSV bronchiolitis had greater amounts of CD11b and MPO than blood neutrophils and BAL fluid neutrophils from controls (p<0.01). Blood neutrophils from term infants with RSV bronchiolitis had less total TLR4 protein than preterm infants with RSV bronchiolitis (p = 0.005), and both had less than controls (p<0.04). Total TLR4 for each group was greater in BAL fluid neutrophils than in blood neutrophils. Blood neutrophils from preterm infants with RSV bronchiolitis had greater TLR4 mRNA expression than term infants with RSV bronchiolitis (p = 0.005) who had similar expression to controls (p = 0.625). CONCLUSIONS: In infants with severe RSV bronchiolitis, neutrophil activation starts in the blood and progresses as they are recruited into the airways. Total neutrophil TLR4 remains low in both compartments. TLR4 mRNA expression is unimpaired. This suggests that neutrophil TLR4 expression is deficient in these infants, which may explain why they develop severe RSV bronchiolitis.
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Bronquiolite Viral/metabolismo , Neutrófilos/metabolismo , Infecções por Vírus Respiratório Sincicial/metabolismo , Vírus Sincicial Respiratório Humano , Receptor 4 Toll-Like/metabolismo , Doença Aguda , Biomarcadores/metabolismo , Bronquiolite Viral/imunologia , Bronquiolite Viral/virologia , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Líquido da Lavagem Broncoalveolar/virologia , Humanos , Lactente , Recém-Nascido , Recém-Nascido Prematuro , Neutrófilos/imunologia , RNA Mensageiro/imunologia , RNA Mensageiro/metabolismo , Infecções por Vírus Respiratório Sincicial/imunologia , Infecções por Vírus Respiratório Sincicial/virologia , Receptor 4 Toll-Like/imunologiaRESUMO
INTRODUCTION: Suboptimal communication between health professionals has been recognised as a significant causative factor in incidents compromising patient safety. The use of a structured method of communication has been suggested to improve the quality of information exchange. The aim of this study was to determine if the teaching of a communication tool, ISBAR (Identify, Situation, Background, Assessment, Recommendation), a modification of SBAR (Situation, Background, Assessment, Recommendation), improved the content and clarity of a telephone referral in an immersive simulated clinical scenario conducted in real time. METHOD: Seventeen teams of final-year medical students were randomised into two groups. The intervention group participated in a 40 min education session about the ISBAR communication tool. A control group received no training. Each team of five students participated in a simulated clinical scenario using a patient simulator in a mocked-up clinical environment. During each scenario, one student made a telephone referral seeking assistance from a senior colleague. Audio data for the telephone referrals (n = 17 students) were captured during the scenario for both groups. During a blinded review of the data, communication was scored on both content and clarity. RESULTS: Communication content was higher from a mean score of 10.2 to 17.4 items (p<0.001) with the intervention. Clarity of the delivery of information on a 5-point scale was also higher in the intervention group (rho = 0.903, p<0.001). CONCLUSIONS: The teaching of a structured method of communication improved the communication during telephone referral in a simulated clinical setting. This research has implications for how healthcare professionals are taught to communicate with each other.
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Comunicação , Relações Interprofissionais , Ensino , Telemedicina , Educação de Graduação em Medicina , Humanos , Encaminhamento e ConsultaRESUMO
AIMS: To determine the effect of carbon sources on cellulose produced by Gluconacetobacter xylinus strain ATCC 53524, and to characterize the purity and structural features of the cellulose produced. METHODS AND RESULTS: Modified Hestrin Schramm medium containing the carbon sources mannitol, glucose, glycerol, fructose, sucrose or galactose were inoculated with Ga. xylinus strain ATCC 53524. Plate counts indicated that all carbon sources supported growth of the strain. Sucrose and glycerol gave the highest cellulose yields of 3.83 and 3.75 g l(-1) respectively after 96 h fermentation, primarily due to a surge in cellulose production in the last 12 h. Mannitol, fructose or glucose resulted in consistent rates of cellulose production and yields of >2.5 g l(-1). Solid state (13)C CP/MAS NMR revealed that irrespective of the carbon source, the cellulose produced by ATCC 53524 was pure and highly crystalline. Scanning electron micrographs illustrated the densely packed network of cellulose fibres within the pellicles and that the different carbon sources did not markedly alter the micro-architecture of the resulting cellulose pellicles. CONCLUSIONS: The production rate of bacterial cellulose by Ga. xylinus (ATCC 53524) was influenced by different carbon sources, but the product formed was indistinguishable in molecular and microscopic features. SIGNIFICANCE AND IMPACT OF THE STUDY: Our studies for the first time examined the influence of different carbon sources on the rate of cellulose production by Ga. xylinus ATCC 53524, and the molecular and microscopic features of the cellulose produced.
Assuntos
Metabolismo dos Carboidratos , Carbono/metabolismo , Celulose/química , Celulose/metabolismo , Gluconacetobacter xylinus/metabolismo , Celulose/ultraestrutura , Contagem de Colônia Microbiana , Meios de Cultura , Fermentação , Frutose , Galactose , Gluconacetobacter xylinus/crescimento & desenvolvimento , Glucose , Glicerol , Manitol , Microscopia Eletrônica de Varredura , Ressonância Magnética Nuclear Biomolecular , SacaroseRESUMO
The complement regulatory proteins CD55 and CD59 are expressed on the plasma membrane of human spermatozoa, whereas CD46 is only on the inner acrosomal membrane (IAM) which becomes surfaced exposed after the acrosome reaction when sperm assume fertilisation-competence. CD55 & CD59, two glycosylphosphatidylinositol (GPI)-anchored proteins, have been detected previously in some studies also in the acrosomal region of chemically fixed spermatozoa but never demonstrated at this site on unfixed spermatozoa. Dual labelling immunofluorescence and confocal microscopy on fresh unfixed spermatozoa, with minimal subsequent time to fixation, has shown CD55 to be markedly expressed on the IAM, more than on the plasma membrane. However, unlike for CD46, CD55 displayed patchy staining over the acrosome, with some variation between individual spermatozoa. All IAM-associated CD55 was localised within GM1-containing lipid rafts. CD59 was expressed also on the IAM, but in a pronounced granular pattern with more variation observed from one spermatozoa to another. Both CD55 & CD59 were released from the IAM by PI-PLC, demonstrating them to be GPI-anchored. Analysis of acrosome-reacted spermatozoal CD55 by Western blotting revealed a novel single 55 kDa protein lacking significant oligosaccharides susceptible to glycosidases. Antibody-induced membrane rafting and release of CD55 & CD59 in vitro may have influenced previous results. Significant coexpression of CD55 & CD46 on the IAM suggests some functional cooperation at this site.
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Acrossomo/imunologia , Antígenos CD/metabolismo , Proteínas Inativadoras do Complemento/metabolismo , Western Blotting , Antígenos CD55/metabolismo , Antígenos CD59/metabolismo , Membrana Celular/imunologia , Humanos , Masculino , Proteína Cofatora de Membrana/metabolismo , Microscopia de Fluorescência , Fosfatidilinositol Diacilglicerol-Liase/imunologia , Fosfoinositídeo Fosfolipase C , Espermatozoides/imunologia , Fatores de Tempo , Preservação de TecidoRESUMO
A methodology for the quick and efficient study of phagocytosis has been developed. It uses the flow cytometer to exploit the change in size and granularity that occurs in cells upon the ingestion of particulate material. The numbers of cells that have phagocytosed particles can be calculated from the distinct shift in regions that occurs. The method also allows the factors governing phagocytosis to be studied in detail through the use of blocking agents or antibodies. Blood-derived monocytes were studied to investigate the role of complement in metal particle phagocytosis to further understand aseptic loosening. Factor C3b was found to be fundamental to the opsonization and phagocytosis of metal particles by monocytes.
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Complemento C3b/metabolismo , Metais/metabolismo , Fagócitos/citologia , Fagócitos/metabolismo , Fagocitose , Anticorpos/imunologia , Quelantes/farmacologia , Fator H do Complemento/farmacologia , Ácido Edético/farmacologia , Fibrinogênio/farmacologia , Citometria de Fluxo , Humanos , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Metais/química , Fagócitos/efeitos dos fármacos , Fagocitose/efeitos dos fármacosRESUMO
Activated mast cells generate multiple cytokines but it is not known if these can be differentially regulated by pharmacological agents. We report here that the glucocorticoid dexamethasone (DEX) preferentially inhibited Ag-induced expression of IL-4 and IL-6 mRNA relative to TNF-alpha mRNA in RBL-2H3 cells. Likewise, the drug more readily inhibited release of IL-4 than TNF-alpha protein. SB203580, an inhibitor of p38 mitogen-activated protein kinase (MAPK), enhanced Ag-induced TNF-alpha mRNA expression without affecting IL-4 or IL-6 mRNA. At the protein level, SB203580 exerted little effect on TNF-alpha release but inhibited IL-4 release; notably, the ratio of TNF-alpha : IL-4 increased markedly with the concentration of SB203580, confirming the differential regulation of these cytokines. PD98059, an inhibitor of MAPK kinase (MEK), a component of the p44/42 MAPK pathway, partially inhibited Ag-induced expression of mRNA for all three cytokines while cyclosporin A inhibited Ag-induced IL-4 and IL-6 mRNA more readily than TNF-alpha mRNA. Ag activation of the cells led to phosphorylation of p38 and p44/42 MAPK but this was not influenced by DEX. In conclusion, mast cell cytokines can be differentially regulated pre- and post-translationally by DEX and SB203580 but there does not appear to be a direct mechanistic link between the actions of these two drugs.
Assuntos
Citocinas/análise , Dexametasona/imunologia , Inibidores Enzimáticos/imunologia , Glucocorticoides/imunologia , Imidazóis/imunologia , Mastócitos/imunologia , Piridinas/imunologia , Células Cultivadas , Ciclosporina/imunologia , Flavonoides/imunologia , Humanos , Interleucina-4/análise , Interleucina-6/análise , Fosforilação , RNA Mensageiro/análise , Serotonina/imunologia , Fator de Necrose Tumoral alfa/análiseRESUMO
Respiratory syncytial virus (RSV) bronchiolitis is an important cause of severe respiratory disease in infants. This study aimed to characterise changes in pulmonary pro- and anti-inflammatory responses in infants with RSV bronchiolitis over the course of the illness. On the day of intubation (Day 1) and the day of extubation (Day X), nonbronchoscopic bronchoalveolar lavage was performed on term and preterm infants ventilated for RSV bronchiolitis and on control infants on Day 1. Tumour necrosis factor (TNF)-alpha, soluble TNF receptor (sTNFR) and interleukin (IL)-6 messenger ribonucleic acid (mRNA) and protein were measured. Twenty-four infants, born at term and 23 infants born preterm with RSV bronchiolitis and 10 controls were recruited. TNF-alpha and IL-6 mRNA and protein in infants with bronchiolitis were greater than the control group on Day 1. In preterm infants, who were ventilated for longer than term infants, TNF-alpha and IL-6 proteins decreased between Day 1 and Day X. Concentrations of sTNFRs differed between groups on Day 1, but levels did not change between Day 1 and Day X. Large amounts of tumour necrosis factor-alpha and interleukin-6 in the respiratory syncytial virus-infected lung suggest important roles for these cytokines in the pathogenesis of respiratory syncytial virus bronchiolitis. The decrease in tumour necrosis factor-alpha and interleukin-6 protein in preterm infants may reflect the prolonged clinical course seen in these infants.
Assuntos
Infecções por Vírus Respiratório Sincicial/fisiopatologia , Bronquiolite Viral/fisiopatologia , Feminino , Humanos , Lactente , Recém-Nascido , Doenças do Prematuro/fisiopatologia , Interleucina-6/análise , Pulmão/fisiopatologia , Masculino , RNA Mensageiro/análise , Receptores do Fator de Necrose Tumoral/análise , Fator de Necrose Tumoral alfa/análiseRESUMO
The androgen receptor gene (AR) contains a domain which includes a variable number of CAG sequences and alleles with low numbers of CAG repeats show high transactivation activity when complexed with testosterone. The ratio of 2nd and 4th digit length (2D:4D) is negatively correlated with phenotypic effects of testosterone. Low numbers of CAG repeats and low 2D:4D are both associated with high sperm numbers and protection against breast cancer. This suggests that CAG number and 2D:4D are correlated i.e. low CAG number and low 2D:4D indicate high activation of androgen-responsive genes. Findings from AR studies predict that low 2D:4D will be associated with prostate and hepatocellular cancer, urolithiasis, ADHD, ankylosing spondylitis, spontaneous abortion, and polycystic ovaries, while high 2D:4D will be associated with motor neuron diseases and endometrial cancer. Findings from 2D:4D studies predict that short CAG length will be common in autism and Asperger's syndrome, while high numbers of CAG repeats will be found in men who are prone to early myocardial infarction.
Assuntos
Dedos/anatomia & histologia , Receptores Androgênicos/genética , Caracteres Sexuais , Repetições de Trinucleotídeos , Neoplasias da Mama/genética , Criança , Transtornos Globais do Desenvolvimento Infantil/genética , Feminino , Fertilidade/genética , Dedos/crescimento & desenvolvimento , Predisposição Genética para Doença , Humanos , Masculino , Infarto do Miocárdio/genética , Fenótipo , Valor Preditivo dos Testes , Gravidez , Estrutura Terciária de Proteína , RiscoRESUMO
Despite four decades of advances in the design of orthopaedic devices aseptic loosening remains a major cause for the revision of total joint arthroplasty. This study used the techniques of immunohistochemistry and reverse transcription polymerase chain reaction to identify the inflammatory cell types, cytokines and chemokines within the interface tissue surrounding failed Accord Knee prostheses. Many T cells were identified within the tissue: however, the classical marker of activation, CD25 was expressed on very few cells. Molecular analysis failed to detect the synthesis of either Th1 or Th2 cytokines. These results suggest that the T cells are being actively recruited to the site of inflammation along the chemokine gradients but are not participating in a classical immune response.