Cerebrospinal fluid xenin levels during body mass reduction: no evidence for obesity-associated defective transport across the blood-brain barrier.

CONTEXT: Recent studies have shown that xenin can act in the hypothalamus, reducing food intake through a leptin- and melanocortin system-independent mechanism. OBJECTIVE: To evaluate the impact of body mass reduction on the blood and cerebrospinal fluid (CSF) levels of xenin. DESIGN AND SETTING: Thirteen obese patients (11 women) selected for roux-in-Y gastric bypass surgery were evaluated before and approximately 8 months after surgery. Xenin was determined in serum and CSF by radioimmunoassay. RESULTS: As compared with lean subjects, obese patients have increased blood levels of xenin, which reduce after surgery. There are significant correlations between blood xenin and blood leptin and insulin levels. CSF concentration of xenin is ∼10-fold lower than blood levels, and is significantly higher in obese subjects as compared with lean ones, returning to normal levels after body mass reduction. There is a significant linear correlation between CSF and blood levels of xenin. CONCLUSION: Xenin is present in the human CSF in a concentration ∼10-fold lower than the blood. Both blood and CSF xenin are correlated with blood levels of important markers of adiposity, leptin and insulin. The levels of CSF xenin are linearly correlated with blood xenin, independently of patient body mass, suggesting that either its transport across the blood-brain barrier is not saturated in the concentration range detected in this study or that there is a coordinated release of xenin from the periphery and the CNS.

Effect of treadmill gait on bone markers and bone mineral density of quadriplegic subjects.

Quadriplegic subjects present extensive muscle mass paralysis which is responsible for the dramatic decrease in bone mass, increasing the risk of bone fractures. There has been much effort to find an efficient treatment to prevent or reverse this significant bone loss. We used 21 male subjects, mean age 31.95 +/- 8.01 years, with chronic quadriplegia, between C4 and C8, to evaluate the effect of treadmill gait training using neuromuscular electrical stimulation, with 30-50% weight relief, on bone mass, comparing individual dual-energy X-ray absorptiometry responses and biochemical markers of bone metabolism. Subjects were divided into gait (N = 11) and control (N = 10) groups. The gait group underwent gait training for 6 months, twice a week, for 20 min, while the control group did not perform gait. Bone mineral density (BMD) of lumbar spine, femoral neck, trochanteric area, and total femur, and biochemical markers (osteocalcin, bone alkaline phosphatase, pyridinoline, and deoxypyridinoline) were measured at the beginning of the study and 6 months later. In the gait group, 81.8% of the subjects presented a significant increase in bone formation and 66.7% also presented a significant decrease of bone resorption markers, whereas 30% of the controls did not present any change in markers and 20% presented an increase in bone formation. Marker results did not always agree with BMD data. Indeed, many individuals with increased bone formation presented a decrease in BMD. Most individuals in the gait group presented an increase in bone formation markers and a decrease in bone resorption markers, suggesting that gait training, even with 30-50% body weight support, was efficient in improving the bone mass of chronic quadriplegics.

Accuracy of the urinary albumin to creatinine ratio as a predictor of albuminuria in adults with sickle cell disease.

AIM: To test the usefulness of a random urine specimen albumin to creatinine ratio (A/C) in predicting 12 hour urinary albumin excretion (12UA) in patients with sickle cell disease. METHODS: 12UA and A/C were measured in nocturnal urine collections and random morning urine samples, respectively, of 72 patients with sickle cell disease. RESULTS: The correlation of A/C values with 12UA values did not provide support for the use of random urine specimens for predicting urinary albumin excretion (UAE) in these patients. However, values of A/C >/= 0.45 and < 0.45 were indicative of raised and normal UAE, respectively. The sensitivity, specificity, and accuracy of the test were 100.0%, 87.2%, and 91.7%, respectively. CONCLUSIONS: This method cannot be recommended for predicting 12UA in patients with sickle cell disease, but it is useful for selecting patients who should collect 12 hour urine for the estimation of UAE.

The relevance of laboratory diagnosis of human cryptosporidiosis and other coccidia.

Human infection by Cryptosporidium spp and other coccidia are due to opportunist non-host specific microorganisms. In HIV seropositive patients, the gastrointestinal symptoms accompanying such infections may be serious and prolonged and may include nausea, low-grade fever, abdominal cramps, anorexia and watery diarrhoea. We studied 188 stool samples from 111 patients (84 men and 27 women) with diarrhoea. A modified Ziehl-Nielsen technique for the detection of Cryptosporidium spp and Isospora belli was employed. The mean age of the patients was 31 years. Cryptosporidium spp was seen in 18% (n = 20) of the patients, 90% (n = 18) of whom were HIV seropositive. Isospora belli was recorded only from HIV seropositive patients (5.4% of all the patients studied and 6.5% of those who were HIV seropositive). These data confirm the good results obtained with this technique for the identification of Cryptosporidium spp and other coccidia and also reaffirm the clinical importance of correctly diagnosing the cause of diarrhoea, particularly in HIV seropositive patients.

Cerebrospinal fluid profiles in acquired immunodeficiency syndrome with and without neurocryptococcosis.

Cryptococcosis is one of the most common fungal infections of the central nervous system (CNS) in AIDS patients and meningoencephalitis or meningitis is a frequently observed manifestation. However, systematic studies of cerebrospinal fluid (CSF) composition from AIDS patients with CNS cryptococcosis have been few. CSF samples from 114 HIV seropositive patients whose clinical complaint suggested CNS involvement, were analyzed; 32 samples from patients diagnosed as having neurocryptococcosis (Group 1) and 82 samples from patients with no identified neurological disfunction (Group 2). Based on cytological and biochemical results, two distinct profiles were observed: Normal (Group 1 = 31%, Group 2 = 39%); Abnormal (Group 1 = 69%, Group 2 = 61%). Lymphocytes were the most frequent cells in both groups. Our CSF cytological and biochemical findings showed that in AIDS patients liquoric abnormalities are quite frequent, non-specific and difficult to interpret. In these circumstances a systematic search to identify the etiologic agent using microbiological and/or immunological assays must be routinely performed.

[Measuring cyclosporine levels in whole blood in kidney transplantation]. / Monitoramento dos níveis de ciclosporina em sangue total em transplantes renais.

BACKGROUND: Cyclosporin A is a potent immunosuppressive drug effective in combatting rejection following organ transplantation. In na effort to replace a radioimmunoassay (RIA) for whole blood determination of cyclosporine (Cya) we compared RIA with fluorescence polarization immunoassay (FPIAm). METHOD: 65 blood samples were analysed from kidney transplanted patients. The samples were collected into tubes containing EDTA as anticoagulant and analysed by RIA and FPIAm. RESULTS: The statistical analysis revealed a difference between both methods (p < 0.05). The linear-regression comparison of Cya concentration measured by RIA and FPIAm showed the following relationship: Cya(FPIAm) = 1.06 x Cya(RIA) + 5.8 (r = 0.9817). CONCLUSION: We conclude that FPIAm provides na alternative method for measuring cyclosporine in whole blood with the added advantages of being reasonably rapid, precise and easy to perform.

UriSed as a screening tool for presumptive diagnosis of urinary tract infection.

BACKGROUND: Although a quantitative urine culture is essential for the final diagnosis of urinary tract infection, it is time-consuming and an expensive procedure. Effective screening tests would be a promising alternative to provide immediate results for the clinician and eliminate unnecessary culturing for most of the negative samples. The aim of this study was to evaluate the performance of an automated sediment analyzer (UriSed) as screening tool for presumptive diagnosis of urinary tract infection. METHODS: We studied 1379 fresh midstream clean-catch urine samples from children to elderly. All samples were submitted to automated sediment analysis (UriSed) and quantitative urine culture (CLED medium agar). RESULTS: The sediment analyzer detected leukocyturia and/or significant bacteriuria with sensitivity of 97%, specificity of 59%, positive predictive value of 27%, negative predictive value of 99%, and accuracy of 64% at cutoff values of bacteria count ≥12.6 elements/hpf and WBC ≥6 cells/hpf. These data suggest a potential 52% reduction of unnecessary urine cultures. CONCLUSION: The UriSed seems to be an efficient tool for screening UTI with high sensitivity and low rate of false-negative results.

Urinálise: comparação entre microscopia óptica e citometria de fluxo / Urinalysis: comparison between microscopic and flow cytometry analysis

INTRODUÇÃO E OBJETIVO: O exame de urina é um procedimento de alta demanda, trabalhoso e pouco padronizado. Este estudo teve por objetivo avaliar o desempenho de um citômetro de fluxo na realização do exame de urina de rotina. CASUíSTICA E MÉTODOS: Foram analisadas 1.140 amostras de urina através de microscopia óptica comum e de citometria de fluxo (UF-100/SYSMEX). A precisão foi estabelecida com a análise de quatro amostras de urina (20 replicações cada). O cálculo da reprodutibilidade foi realizado a partir de 30 determinações de dois controles comerciais em dias consecutivos. RESULTADOS: As contagens de hemácias e leucócitos mostraram concordância de 91 por cento e 93 por cento, respectivamente. Cilindros, células e bactérias mostraram sobreposição dos valores fornecidos pelo UF-100 quando comparados com os relatados na análise microscópica. A precisão do UF-100 variou de 4 por cento a 155 por cento, com reprodutibilidade de 3 por cento e 25 por cento, dependendo do parâmetro avaliado. CONCLUSÃO: O equipamento UF-100/SYSMEX demonstra boa precisão, reprodutibilidade e concordância com a microscopia óptica. A utilização da citometria de fluxo implica numa maior agilização e padronização da rotina, bem como em uma nova maneira de reportar e interpretar o exame de urina de rotina.

INTRODUCTION: Urinalysis is a high demand procedure, with large amount of manual labor and poorly standardized. The purpose of this investigation was to analyze the performance of an automated system based on flow cytometry for routine urinalysis. MATERIAL AND METHODS: We analyzed 1,140 urine samples by light field microscopy and by flow cytometry (UF-100/SYSMEX). For the precision study of the UF-100, we calculated the within-run and between-run coefficients of variation using two different levels of commercial controls and four different urine samples. RESULTS: Erythrocytes and leukocytes counts by the two methods showed an agreement of 91 percent and 93 percent, respectively. Casts, epithelial cells and bacteria counts by the UF-100 showed a significant overlap when compared to microscopic analysis. Intra assay precision (within-run) ranged from 4 percent to 155 percent and interassay precision (between-run) varied from 3 percent to 25 percent, depending on the considered parameter. CONCLUSION: Flow cytometry is a precise and reproducible technique, with a strong correlation with the results obtained by microscopic analysis. Flow cytometry allows a better workflow and a new manner of reporting and interpreting routine urinalysis.

Effect of treadmill gait on bone markers and bone mineral density of quadriplegic subjects

Quadriplegic subjects present extensive muscle mass paralysis which is responsible for the dramatic decrease in bone mass, increasing the risk of bone fractures. There has been much effort to find an efficient treatment to prevent or reverse this significant bone loss. We used 21 male subjects, mean age 31.95 ± 8.01 years, with chronic quadriplegia, between C4 and C8, to evaluate the effect of treadmill gait training using neuromuscular electrical stimulation, with 30-50 percent weight relief, on bone mass, comparing individual dual-energy X-ray absorptiometry responses and biochemical markers of bone metabolism. Subjects were divided into gait (N = 11) and control (N = 10) groups. The gait group underwent gait training for 6 months, twice a week, for 20 min, while the control group did not perform gait. Bone mineral density (BMD) of lumbar spine, femoral neck, trochanteric area, and total femur, and biochemical markers (osteocalcin, bone alkaline phosphatase, pyridinoline, and deoxypyridinoline) were measured at the beginning of the study and 6 months later. In the gait group, 81.8 percent of the subjects presented a significant increase in bone formation and 66.7 percent also presented a significant decrease of bone resorption markers, whereas 30 percent of the controls did not present any change in markers and 20 percent presented an increase in bone formation. Marker results did not always agree with BMD data. Indeed, many individuals with increased bone formation presented a decrease in BMD. Most individuals in the gait group presented an increase in bone formation markers and a decrease in bone resorption markers, suggesting that gait training, even with 30-50 percent body weight support, was efficient in improving the bone mass of chronic quadriplegics.

The relevance of laboratory diagnosis of human cryptosporidiosis and other coccidia

Infeccao humana por Cryptosporidium spp e outros coccideos deve-se a microrganismos oportunistas hospedeiros nao-especificos e, em pacientes HIV positivos, os sintomas gastro-intestinais podem ser serios e prolongados incluindo nauseas, febre baixa, dores abdominais, anorexia e diarreia aquosa. Estudamos 188 amostras de fezes provenientes de 111 pacientes (84 homens e 27 mulheres, com idade media de 31 anos) com diarreia atendidos no Hospital das Clinicas da Universidade Estadual de Campinas. Para deteccao de Cryptosporidium spp e Isospora belli empregou-se a tecnica de Ziehl-Nielsen modificada. 83 por cento dos pacientes (n=92) eram HIV positivos. Cryptosporidium spp foi observado em 18 por cento de todos os pacientes, sendo que destes, 90 por cento eram HIV positivos. Isospora belli foi evidenciada somente em pacientes HIV positivos, com uma frequencia de 5,4 por cento de todos os pacientes e de 6,5 por cento considerando-se apenas os pacientes HIV positivos...

Monitoramento dos níveis de ciclosporina em sangue total en transplantes renais / Measuring cyclosporine in whole blood in kidney transplantation176-8

Introduçao e Objetivos. Ciclosporina A é uma droga imunossupressora potente e efetiva no combate à rejeiçao de órgaos transplantados. No presente estudo, os autores avaliaram o emprego de um imunoensaio monoclonal com fluorescência polarizada (FPIAm), como um método alternativo ao radioimunoensaio (RIA) para determinaçao dos níveis de ciclosporina em sangue total. Material e Métodos. Amostras de sangue de 65 pacientes submetidos a transplante renal foram colhidas em frascos com EDTA 12 horas após a última dose de ciclosporina, via oral. Os níveis de cilcosporina foram avaliados por meio de radioimunoensaio com anticorpo monoclonal e imunoensaio monoclonal com fluorescência polarizada. Resultados e Conclusao. A análise estatística revelou um coeficiente de correlaçao entre os métodos de r = 0,9817 e o teste t de Student pareado mostrou haver diferença estatisticamente significante entre eles (p<0,05). A análise da regressao revelou que os métodos poderiam ser comparáveis por meio da equaçao Cya(FPIAm) = 1,06xCya(RIA) + 5,8, mostrando que FPIAm é um excelente método alternativo ao RIA, com as vantagens de ser rápido, de fácil execuçao, reprodutível e com resultados comparavéis aos obtidos por RIA.

A acetilacao da isoniazida em paulistas descendentes de japoneses. / Isoniazid acetylation in Japanese descendants born in the State of Sao Paulo

O fenotipo acetilador da isoniazida (INH) foi investigado em 88 paulistas descendentes nao miscigenados de japoneses internados para tratamento de tuberculose em sanatorios de Campos do Jordao,SP. Para tanto, investigou-se o percentual de acetilisoniazida presente na urina 10 horas apos a ingestao de 10mg de INH por kg de peso corporal. Os resultados obtidos permitiram estimar a frequencia de acetiladores lentos da isoniazida entre os paulistas descendentes de japoneses nao miscigenados em aproximadamente, 10%, frequencia essa muito semelhante a observada no Japao

Comparaçåo entre radioimunoensaio e imunoturbidimetria para determinaçåo da microalbumina / A comparasion of radioimmunoassay and immunoturbidimetry for determination of microalbuminuria

Microalbuminuria é a excreçåo urinária de pequenas quantidades de albumina, insuficiente para serem detectadas por métodos usais de dosagem de proteínas e possui valor preditivo para o desenvolvimento de proteinúria e insuficiência renal crônica. Radioimunoensaio com duplo anticorpo (RIA) é usualmente considerado como um método-padråo para esta dosagem. Com o objetivo de comparar as dosagens de albumina urinária por radioimunoensaio e por imunoturbidimetria (IT), foram analisadas 91 amostras de urina de pacientes diabéticos ou nåo. Verificamos nåo haver diferença estatisticamente significativa entre os valores obtidos por ambos os métodos e um coeficiente de correlaçåo r=0,99874, mostrando que a IT pode ser utilizada como um método alternativo ao RIA

Determinacao do fenotipo de acetilacao da isoniazida em lupus eritematoso sistemico. / Determination of the acetylator phenotype of isoniazid in systemic lupus erythematosus

O fenotipo de acetilacao da isoniazida foi determinado em 20 pacientes com lupus eritematoso sistemico (LES). Dez pacientes (50%) eram acetiladores lentos de dez outros acetiladores rapidos. Nao houve correlacao do fenotipo acetilador e manifestacoes clinicas, padrao de fator antinucleo ou alteracoes laboratoriais. O fenotipo acetilador lento nao parece ser suficiente para faciliar a expressao clinica do LES