Distinct inflammatory gene expression in extraocular muscle and fat from patients with Graves' orbitopathy.

OBJECTIVE: This study sought to compare patients with thyroid eye disease (TED) and normal controls with respect to the expression of the NR3C1, CHUK, IKBKB, FOS, NFKB and HSD11B1 genes in orbital fat (OF) and extraocular muscle (EOM). DESIGN AND METHODS: A prospective study design was used to evaluate 34 TED patients and 38 healthy controls. OF was harvested from 33 TED patients and 27 controls. EOM biopsies were obtained from 32 TED patients and 18 controls. Samples were examined by real-time PCR and evaluated using appropriate statistical analyses with a significance cut-off of P < 0.05. RESULTS: NR3C1 mRNA levels were higher in TED EOM (median 213 (96-376)) than those in control EOM (78 (34-138)) (P < 0.001), and NFKB expression was elevated in TED muscle (223 (31-520)) relative to that in control muscle (8 (6-31)) (P < 0.001). HSD11B1 expression was higher in TED EOM (0.78 (0.47-2.01)) than that in control EOM (0.22 (0.09-0.51)) (P < 0.001). Levels of CHUK, IKBKB, and FOS were higher in TED EOM (115 (20-223), 111 (54-299) and 0.11 (0.03-0.19), respectively) than those in control EOM (5.8 (2-13), 21 (5-52) and 0.05 (0.001-0.03) respectively) (P < 0.001). CONCLUSION: Tissues involved in GO exhibited different mRNA levels of NR3C1, CHUK, IKBKB, FOS, NFKB and HSD11B1. Gene expression in OF was similar for TED patients and controls. CHUK, IKBKB, FOS, NFKB, and HSD11B1 mRNA levels were higher in TED EOM than those in control EOM. NFKB was disproportionally elevated compared with NR3C1; this finding was indicative of a local proinflammatory profile.

Correlates of human immunodeficiency virus cervicovaginal shedding among postmenopausal and fertile-aged women.

OBJECTIVE: The aims of this study were to compare the intensity of human immunodeficiency virus (HIV)-RNA genital shedding among postmenopausal (PM) and fertile-aged (F) women and to investigate the association between viral shedding and gynecological features, HIV plasma viral loads, and other markers of HIV disease progression. METHODS: We interviewed 146 HIV-infected women (73 PM/73 F) in search of gynecological complaints and signs and symptoms of HIV disease and obtained additional information concerning HIV infection by medical chart review. Cervicovaginal lavages (CVLs) were collected for assessment of HIV shedding. Laboratory analyses included CD4 cell counts, HIV-RNA quantitation in plasma and CVL, and screening for concurrent genital infections. RESULTS: HIV-RNA genital shedding was detected in 16.4% of PM and 21.9% of F women (P = 0.400), and the intensity of HIV shedding did not differ between both groups (means-PM: 1.4log/mL; F: 1.4log/mL; P = 0.587). Three women (2 PM/1 F) exhibited viral shedding in the absence of detectable viremia. HIV plasma viral loads correlated with HIV shedding in both groups. In multivariable analysis, HIV plasma viral loads were independently associated with HIV shedding in both groups. Moreover, the intensity of shedding was independently associated with vaginal pH, tumor necrosis factor α concentrations in CVL, and HIV plasma viral loads. CONCLUSIONS: Despite significant changes that occur in the vaginal mucosa of PM women, HIV cervicovaginal shedding was not significantly influenced by this state in our cohort. In contrast, increased vaginal pH and genital inflammation, evidenced by increased tumor necrosis factor α concentrations in CVL and HIV plasma viral loads, were independently associated with the intensity of HIV shedding in PM and F women.

Avaliação da excreção genital do HIV-1 em mulheres menopausadas e em idade fértil: prevalência e fatores associados / HIV cervicovaginal shedding among postmenopausal and fertile-aged women: prevalence and associated factors

INTRODUÇÃO: Poucos estudos têm focado as modificações fisiológicas que ocorrem no trato genital de mulheres menopausadas infectadas pelo HIV e sua associação com a excreção genital do vírus. Nesse estudo de corte transversal, comparou-se a excreção genital do HIV em mulheres menopausadas e em idade fértil em acompanhamento em um centro especializado em São Paulo, Brasil. Investigou-se também a associação entre a excreção genital de RNA de HIV e a viremia em ambos os grupos. Fatores associados com a intensidade da excreção genital de HIV também foram pesquisados, incluindo achados ginecológicos e marcadores de progressão da infecção por HIV. MÉTODOS: 146 mulheres infectadas pelo HIV [73 menopausadas (M)/73 em idade fértil (F)] foram selecionadas em Serviço de Extensão ao Atendimento de Pacientes com HIV/Aids Casa da Aids do Hospital das Clínicas da FMUSP, São Paulo, Brasil. As mulheres menopausadas referiram tempo médio de 8,17 anos (DP=6 anos) de menopausa. A contagem de linfócitos T CD4+ foi obtida por citometria de fluxo e a quantificação do RNA do HIV no plasma e no lavado cervicovaginal (LCV) foi realizada por RT-PCR quantitativo, utilizando-se o kit Cobas Amplicor HIV-1 Monitor Test®, no método ultrasensível. Cloreto de lítio foi introduzido no tampão para obtenção do LCV e quantificado antes e depois da coleta do lavado, a fim de determinar o fator de diluição de cada amostra. A deteção do gene SRY por PCR também foi realizada a fim de eliminar amostras com eventual contaminação espermática. A prevalência de excreção genital foi estimada para ambos os grupos e os fatores associados à intensidade da excreção viral foram investigados, utilizando-se modelo de regressão linear múltipla. As variáveis com p<0,2 na análise bivariada foram incluídas na análise multivariada, assim como o grupo em estudo (M ou F). O modelo final incluiu fatores que se mostraram independentemente associados com a intensidade da excreção genital de HIV...

BACKGROUND: Few studies have focused on physiological modifications that occur in the genital tract of HIV-infected postmenopausal women and their association with HIV cervicovaginal shedding. In this cross-sectional study we evaluated and compared HIV genital shedding among postmenopausal and fertile-aged women under care at a specialized center in Sao Paulo, Brazil, investigating the association between HIV-RNA shedding and HIV plasma viral loads in both groups. Factors associated with higher HIV shedding were also investigated, including gynaecological features and HIV disease progression markers. METHODS: 146 women living with HIV [73 postmenopausal (PM)/73 in fertile-aged (F)] were enrolled at the HIV Clinic, University of São Paulo Medical School, Brazil. Postmenopausal women referred a mean duration of 8.17y (SD=6y) since menopause. CD4+ cell counts were obtained by flow cytometry and HIV-RNA was quantified in plasma and in cervicovaginal lavages (CVL) by RT-PCR, using Cobas Amplicor HIV-1 Monitor Ultrasensitive Test. Lithium chloride was introduced into the CVL buffer and measured before and after CVL collection in order to determine the dilution factor for each specimen. SRY gene detection by PCR was also performed in all samples in order to rule out sperm contamination. Prevalence of HIV genital shedding was estimated for both groups and factors associated with the intensity of viral shedding were investigated, using a multiple linear regression model. Variables with p<0.2 in bivariate analysis were included in multivariate analysis, as well as the study group (PM and F). The final model included factors shown to be independently associated with intensity of HIV genital shedding. RESULTS: The prevalence of HIV-RNA genital shedding was similar in both groups. (PM: 17.8%, 95%CI 9.8 28.5; F: 22%, 95%CI 13.1 33.1, p=0.678). Likewise, the intensity of HIV shedding was shown not to differ between PM and F women...

Utilidade de regras booleanas aplicadas à liberação de resultados de exames hormonais e marcadores tumorais / Usefulness of Boolean rules applied on the release of hormonal and tumor markers tests results

INTRODUÇÃO: A avaliação de perfis hormonais exige, tradicionalmente, a participação de especialistas para adequada liberação setorial. A análise desses resultados através de regras booleanas é alternativa interessante para minimizar os resultados a serem avaliados manualmente. MATERIAL E MÉTODO: Avaliamos a utilização de regras booleanas construídas no programa Instrument Manager para o equipamento Architect, principalmente para mensuração de hormônios sexuais e tireoidianos, além de marcadores tumorais. O resultado da intervenção foi avaliado quanto a: a) número e facilidade de construção das regras; b) comparação cega entre liberação por patologista clínico (resultados impressos) e conjunto de regras em 940 testes consecutivos. RESULTADOS: A criação das regras se revelou tarefa complexa e trabalhosa, especialmente pela existência de perfis hormonais com diversos modos de solicitação dos testes. Foram necessárias 153 regras booleanas (do tipo se...então) em uma ordem específica. Essas regras concordaram com o especialista em 97,9 por cento dos casos (920 testes). O conjunto de regras reteve 25 casos (2,7 por cento) e o patologista clínico, apenas nove. Nos 20 casos discordantes, as regras deixaram de reter apenas dois casos, sendo um da fração beta da gonadotrofina coriônica humana (beta-hCG) em homens (que implicou a criação de nova regra) e um caso de perfil tireoidiano completo sem hormônio tireoestimulante (TSH) (o patologista clínico optou por confirmar o pedido médico). CONCLUSÃO: A criação de conjunto eficiente de regras booleanas é tarefa complexa que necessita de conhecimentos técnicos e de lógica, mas que permite a otimização do funcionamento do laboratório. Obtivemos excelente concordância entre o conjunto de regras criado e a liberação manual de patologista clínico, garantindo a segurança, a velocidade e o menor custo do sistema.

BACKGROUND: Hormone profile evaluation traditionally requires an expert review of results for adequate laboratory section test release. Analysis of these results using Boolean rules is an interesting alternative to reduce the number of results that require manual review. MATERIAL AND METHOD: We evaluated the utilization of Boolean rules using Instrument Manager software and Architect analyzer, mainly performing sex and thyroid hormones measurement. The intervention was evaluated on: a) number of rules and its easiness of construction; b) blind comparison of results evaluation by clinical pathologist (printed results) and set of rules in 940 consecutive tests. RESULTS: Rule creation was a complex and arduous task, especially due to hormonal profiles with several different request patterns. It was necessary to use a set of 153 Boolean (if...then) rules, in a specific order. This set of rules agreed with expert opinion in 97.9 percent (920 tests). Rules hold 25 tests (2.7 percent) and the clinical pathologist only nine tests. There was discordance in 20 cases; rules did not hold only two cases: a beta-hCG in a male patient (that prompted the creation of a new rule) and a complete thyroid profile lacking only TSH request (pathologist opted to review the original request). CONCLUSION: Creation of an efficient set of Boolean rules proved to be a complex task requiring both technical and logics knowledge, but allowing optimization of laboratory workload. We achieved excellent concordance between the set of rules and clinical pathologist manual review, in a safe, fast and low cost system.