The use of 16s rDNA restriction fragment length polymorphism analysis for the identification of non-fermenting gram negative bacilli in a cystic fibrosis microbiology referral service

The utility of 16s rDNA restriction fragment length polymorphism (RFLP) analysis for the partial genomovar differentiation of Burkholderia cepacia complex bacterium is well documented. We compared the 16s rDNA RFLP signatures for a number of non-fermenting gram negative bacilli (NF GNB) LMG control strains and clinical isolates pertaining to the genera Burkholderia, Pseudomonas, Achromobacter (Alcaligenes), Ralstonia, Stenotrophomonas and Pandoraea. A collection of 24 control strain (LMG) and 25 clinical isolates were included in the study. Using conventional PCR, a 1.2 kbp 16s rDNA fragment was generated for each organism. Following restriction digestion and electrophoresis, each clinical isolate RFLP signature was compared to those of the control strain panel. Nineteen different RFLP signatures were detected from the 28 control strains included in the study. TwentyoneyTwenty- five of the clinical isolates could be classified by RFLP analysis into a single genus and species when compared to the patterns produced by the control strain panel. Four clinical B. pseudomallei isolates produced RFLP signatures which were indistinguishable from B. cepacia genomovars I, III and VIII. The identity of these four isolates were confirmed using B. pseudomallei specific PCR. 16s rDNA RFLP analysis can be a useful identification strategy when applied to NF GNB, particularly for those which exhibit colistin sulfate resistance. The use of this molecular based methodology has proved very useful in the setting of a CF referral laboratory particularly when utilised in conjunction with B. cepacia complex and genomovar specific PCR techniques. Species specific PCR or sequence analysis should be considered for selected isolates; especially where discrepancies between epidemiology, phenotypic and genotypic characteristics occur.

Differences in lung function, growth parameters and frequency of hospital admissions between adolescents with cystic fibrosis-related diabetes and controls

As survival of patients with CF increases,glucose intolerance and cystic fibrosisrelated diabetes (CFRD),ar e increasingly recognised common complications. CFRD may be preceded by a pre-diabetic state. Using markers identified as being associated with CFRD may improve targeted screening. Aim: To identify features consistently predicting CFRD in paediatric patients. Patients diagnosed with CFRD between January 1997–January 2002 were compared with age and sex matched controls. Clinical,micr obiological, and hospitalisation data was collected at time of CFRD diagnosis,and at six monthly intervals for 3 yr prior to diagnosis. Eight patients with CFRD were identified,mean age 13.7 yr (S.D. 3.49) at time of diagnosis. Control patients underwent OGTT to ensure normal glucose tolerance. Patients with CFRD had a lower FEV1 up to 12 months prior to diagnosis however, this was only significant at diagnosis. There was no difference in weight and height z scores between the 2 groups; however,the decrease in weight and height z scores in the CFRD group over 3 yr prior to diagnosis was significant. Mean number of days in hospital and admissions per patient significantly increased in the CFRD group,6 months prior to diagnosis. No other significant differences were observed between the 2 groups. Conclusions: This study has shown a difference in lung function,gr owth parameters and frequency of hospital admissions between patients with CFRD and controls. These differences may be utilised as tools for targeted screening in the paediatricyadolescent population. Further larger scale studies are required to improve guidelines for targeted screening in this population.

Gender differences in CF phenotype associated with low bone mineral density

Adolescents and adults with CF have lower bone mineral density (BMD) than normal, but its relationship with phenotype is not well understood. Point FEV1% predicted (FEV) and rate of change of FEV are biased estimates of disease severity, because progressively older subjects represent a selected survivor population, with females at greater risk of death than males. To investigate the relationship between BMD and phenotype we used an index (predicted age at death) derived from Bayesian estimates of slope and intercept of FEV, age at last measurement and survival status. Predictive equations for the index were derived from 97 subjects (78 survivors) from the RCH CF clinic, and applied to a group of 102 comparable subjects who had BMD measured, classified as having‘mild’ ()75th), ‘moderate’ (25– 75th), or ‘severe’ (-25th centile) phenotype. Total body (TB) and lumbar spine (LS) BMD z-scores (Z) were compared, adjustingfor gender effects, using 2-way ANOVA. Annual mean change in FEV segregated, as expected, according to phenotype, ‘severe’ (ns25), ‘moderate’ (ns51) and ‘mild’ (ns25) y3.01(y3.73 to y2.30)%, y0.85(y1.36 to y0.35)%, 2.70(1.92 to 3.46)%, respectively, with no gender difference. LS and TB BMDZ were different in each phenotype (P-s 0.002), LS BMDZ for ‘severe’, ‘moderate’ and ‘mild’ y1.63(CI: y2.07 to y 1.19), y0.86(CI: y1.17 to y0.55), y0.06(CI: y0.54 to 0.41). Males had lower LS BMDZ than females overall (y1.22 (CI: y1.54 to y0.91) vs. y0.48(CI: y 0.84 to y0.12) Ps0.002). In the ‘severe’ group, males had lower TB BMDZ and LS BMDZ (PF0.002). Low BMD is associated with ‘moderate’ and ‘severe’ phenotypes, with relative preservation in females in the ‘severe’ group. Female biology (reproductive fitness) might promote resistance to bone resorption at a critical level of BMD loss.

Paralisia facial periférica bilateral na leucemia linfóide aguda: relato de caso

A mímica facial é fundamental para a expressão e comunicação humana, que são possíveis apenas através da integridade do nervo facial. Sendo assim, a paralisia facial periférica (PFP) pode deixar seqüelas estéticas, funcionais e psicológicas. A causa mais comum é a paralisia de Bell (50 a 80%), onde a maioria dos pacientes apresenta manifestação unilateral. O acometimento bilateral simultâneo é raro, sendo a leucemia a neoplasia que com maior freqüência pode resultar nesse tipo de manifestação. A seguir, relatamos o caso de um paciente de dezoito anos de idade apresentando leucemia linfóide aguda (LLA) e PFP simultânea, ambas refratárias ao tratamento quimioterápico, culminando com o óbito cinco meses após o início da PFP. Realizou-se considerações importantes sobre a fisiopatologia da PFP na LLA, além de uma revisão da literatura.

Fisioterapia na paralisia facial periférica: estudo retrospectivo

A paralisia facial periférica requer tratamento especializado. A fisioterapia tem como objetivo restabelecer a mímica facial. O objetivo deste estudo foi descrever e analisar os resultados da fisioterapia para indivíduos com paralisia facial periférica. FORMA DE ESTUDO: Retrospectivo. MÉTODO: Foi realizado um estudo retrospectivo em um Hospital Universitário, com autorização do Serviço de Atendimento Médico e Estatístico, no período de 1999 a 2003. Os dados são apresentados em forma descritiva, com utilização de média e mediana para variáveis numéricas e freqüência para variáveis categóricas. RESULTADOS: Foram analisados 23 prontuários durante quatro anos. Foi identificado o predomínio do sexo feminino e a média de idade foi de 32,3 anos (DP±16,5); 14 casos idiopáticas e cinco traumáticas; 12, com comprometimento motor total e 11, parcial; nos 12 casos com avaliação final, sete evoluíram para recuperação parcial e cinco para total. A fisioterapia utilizada foi cinesioterapia e orientações. CONCLUSÃO: Neste estudo os indivíduos são similares a outras populações. Foram tratados com cinesioterapia, como sugerido pela literatura científica e evoluíram com recuperação.

Alcoolismo: efeitos no aparelho vestíbulo-coclear

Diversas drogas ototóxicas causam efeitos nocivos ao ser humano, como zumbido, várias formas de deficiência auditiva e vertigem. Dentre os principais agentes considerados ototóxicos, encontra-se o álcool. OBJETIVO: Verificar as influências do alcoolismo no aparelho vestíbulo-coclear. FORMA DE ESTUDO: Estudo de coorte contemporânea com corte transversal. MATERIAL E MÉTODO: A amostra compreendeu 37 indivíduos para o grupo experimental, freqüentadores do grupo de Alcoólicos Anônimos da cidade de Santa Maria-RS e 37 indivíduos não alcoólicos, com a mesma idade e sexo, de cada indivíduo do estudo, a fim de formar o grupo controle. Todos foram submetidos à anamnese, exame otorrinolarigológico, avaliação audiológica básica e avalição vectoeletronistagmográfica. RESULTADOS: Observou-se que 67,57% dos indivíduos do Grupo Experimental apresentaram alteração na audiometria e 24,32% apresentaram alteração na vectoeletronistagmografia computadorizada. Nos indivíduos do grupo Controle, 27,03% apresentaram audiometria alterada e 10,81% apresentaram resultados alterados na vectoeletronistagmografia computadorizada. CONCLUSÃO: O álcool interfere na audição e no equilíbrio do indivíduo, causando efeito deletério no organismo humano.