JumpSTART Triage Protocol in Disaster Pediatric Patients: A Systematic Literature Review.

INTRODUCTION: In a mass-casualty incident (MCI) involving children, there is a need to apply accurate triage tools in order to help those who require important care, and at the same time, to avoid unnecessary use of resources. Thus, it is discussed which would be the best triage device to use in these situations. One of the most used is a modification of Simple Triage and Rapid Treatment, JumpSTART, whose performative quality this review focuses on. STUDY OBJECTIVE: This review sought to compare the performance parameters of JumpSTART with other triage algorithms used in pediatric disaster victims. METHODS: This systematic review was performed according to the recommendations of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) and registered with the PROSPERO database of systematic reviews with the number CRD42021258415. The last update of the search in the databases was on August 12, 2021 and resulted in six documents to be analyzed. The inclusion criteria included the peer-reviewed academic papers in English, Portuguese, Spanish, and Italian languages, and the databases used were PubMed, Scopus, MEDLINE/Bireme (Virtual Library of Health), Web of Science, and CINAHL, which executes the query on the topic, keywords, or abstracts. Also to be included, documents that were available with full-text access through CAPES, Google, or Google Scholar. Books, non-academic research, and content in languages other than the presented ones were represented as exclusion criteria. The Joanna Briggs Institute (JBI) checklists were used to evaluate the methodological quality of the retrieved studies. The results were presented through narrative synthesis. This review was not funded. RESULTS: Of the collected publications, five articles were used to carry out this review, with the addition of an extra article captured by citation tracking. The findings from the obtained results were that JumpSTART was the preferred tool and presented the fastest speed of use. Only one of the five studies that dealt with accuracy showed JumpSTART as the most accurate algorithm, while three of the other four showed its inferiority in most aspects. In one study, no significant difference was observed amongst the chosen protocols. CONCLUSIONS: There is insufficient evidence to validate JumpSTART as a universal triage tool, given the disparities in the results obtained from the comparisons. No tool performed satisfactorily well, therefore there is an urgent need to create a reliable algorithm.

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.

Subjective qualitative assessment of the finish line of prosthetic preparations submitted to different finishing instruments.

STATEMENT OF PROBLEM: Different instruments have been developed to improve crown finish lines after conventional preparation with diamond rotary instruments. However, more evidence on the effectiveness of these instruments is required. PURPOSE OF STUDY: The purpose of this in vitro study was to assess the effectiveness of 4 different instruments used for cervical margin finishing in regard to the external line angle (ELA). MATERIAL AND METHODS: Forty-eight freshly extracted third molar teeth with similar dimensions were conventionally prepared for crowns and randomly allocated into 4 groups (n=12) according to the finishing instrument: hand trimmer (HT), tungsten carbide bur (TCB), ultrasonic instrument (UI), and diamond rotary instrument (DRI) mounted in a contra angle handpiece with reduced speed. Photomicrographs from the margin profiles of each tooth were obtained with scanning electron microscopy (×40) before and after finishing procedures. The images were qualitatively analyzed (scores 1-4) by 9 experienced dental professionals. Average results were assessed with the paired t test and repeated measures ANOVA followed by Tukey HSD test (α=.05). RESULTS: HT, TCB, and UI demonstrated no significant differences between unfinished and finished mean ELA scores. DRI was the only group to demonstrate a significantly improved ELA (P<.001). Except for HT in relation to TCB (P=.989), all the other groups demonstrated statistically significant different ELA scores. CONCLUSION: DRI mounted in a contra angle handpiece was the only finishing instrument capable of significantly improving the ELA, while TCB and UI produced nonsignificant improvement, and HT was detrimental to the ELA.

Deferasirox in patients with iron overload secondary to hereditary hemochromatosis: results of a 1-yr Phase 2 study.

This open-label, prospective, phase 2 study evaluated the safety and efficacy of deferasirox (10 ± 5 mg/kg/d) in patients with hereditary hemochromatosis (HH) and iron overload refractory to or intolerant of phlebotomy. Ten patients were enrolled and all completed the 12-month treatment period. There were significant decreases from baseline to end of study (i.e., 12 months) in median serum ferritin (P < 0.001), mean transferrin saturation (P < 0.05), median liver iron concentration (P < 0.001), and mean alanine aminotransferase (P < 0.05). The median time to achieve serum ferritin reduction ≥50% compared to baseline was 7.53 months. The most common adverse events were mild, transient diarrhea (n = 5) and nausea (n = 2). No patient experienced an increase in serum creatinine that exceeded the upper limit of normal. These data confirm that deferasirox was well tolerated and effective in reducing iron burden in patients with hereditary hemochromatosis and could be a safe alternative to phlebotomy in selected patients.

Reabilitação de maxila edêntula com prótese fixa implantossuportada: relato de caso / Rehabilitation of edentulous maxilla with implant-supported fixed prosthesis: a case report

Introdução: os problemas funcionais, estéticos e psicológicos decorrentes do edentulismo têm levado a uma busca crescente por reabilitação bucal com próteses sobre implantes. Esse tipo de reabilitação necessita de um planejamento meticuloso, especialmente para os casos de maxila edêntula com prótese fixa sobre implantes, em decorrência da anatomia dessa região. Objetivo: o objetivo do presente caso clínico é descrever a reabilitação de uma maxila edêntula com prótese total fixa implantossuportada, cuja infraestrutura recebeu elementos metalocerâmicos cimentados individualmente.

Introduction: Functional, aesthetic and psychological issues arising from edentulism have led to a growing search for oral rehabilitation with implant-supported prostheses. This type of rehabilitation requires detailed planning, especially for treating edentulous maxilla with implantsupported fixed prostheses due to the anatomy of this region. Objective: The aim of this case report is to describe the rehabilitation of an edentulous maxilla with implantsupported full fixed prosthesis of which framework received metal-ceramic elements individually cemented.

BRAF mutation in the elderly submitted to thyroidectomy.

OBJECTIVE: To evaluate the frequency of the BRAF V600E mutation in patients over 65 years of age undergoing thyroidectomy, correlating its presence or absence with the different histologic lesions, their variants and with prognostic factors of papillary carcinoma. METHODS: We evaluated 85 patients over 65 years of age who underwent thyroidectomy, analyzing the BRAF V600E mutation by RT-PCR performed after DNA extraction from the paraffin blocks. RESULTS: The study detected the presence or absence of BRAF V600E mutation in 47 patients (55.3%). Among the 17 papillary carcinomas studied, seven had the mutation (41.2%). There was a statistical association between the presence of this mutation and the classic variant of papillary carcinoma, and a trend of association with thyroid extravasation. CONCLUSION: BRAF mutation in the elderly is also exclusive of papillary carcinoma and is often significant. Furthermore, it is related to the classic variant and possibly to thyroid extravasation.

Mutação BRAF em pacientes idosos submetidos à tireoidectomia / BRAF mutation in the elderly submitted to thyroidectomy

OBJETIVO: Avaliar a frequência da mutação V600E do gene BRAF em pacientes com mais de 65 anos de idade submetidos à tireoidectomia, correlacionando sua presença ou ausência com as diferentes lesões histológicas, com as variantes e com fatores prognósticos do carcinoma papilífero. MÉTODOS: Foram avaliados 85 pacientes com mais de 65 anos de idade submetidos à tireoidectomia, analisando a mutação BRAF V600E através de reação de PCR-RT realizada após a extração do DNA dos blocos de parafina. RESULTADOS: Detectou-se ausência ou presença da mutação BRAF V600E em 47 pacientes (55,3%). Entre os 17 carcinomas papilíferos estudados, sete apresentavam a mutação (41,2%). Demonstrou-se associação estatística entre a presença desta mutação e a variante clássica do carcinoma papilífero, além de tendência de associação com o extravasamento tireoideano. CONCLUSÃO: A mutação BRAF nos pacientes idosos também é exclusiva do carcinoma papilífero e tem frequência expressiva. Além disso, está relacionada à variante clássica e, possivelmente, ao extravasamento tireoideano.

OBJECTIVE: To evaluate the frequency of the BRAF V600E mutation in patients over 65 years of age undergoing thyroidectomy, correlating its presence or absence with the different histologic lesions, their variants and with prognostic factors of papillary carcinoma. METHODS: We evaluated 85 patients over 65 years of age who underwent thyroidectomy, analyzing the BRAF V600E mutation by RT-PCR performed after DNA extraction from the paraffin blocks. RESULTS: The study detected the presence or absence of BRAF V600E mutation in 47 patients (55.3%). Among the 17 papillary carcinomas studied, seven had the mutation (41.2%). There was a statistical association between the presence of this mutation and the classic variant of papillary carcinoma, and a trend of association with thyroid extravasation. CONCLUSION: BRAF mutation in the elderly is also exclusive of papillary carcinoma and is often significant. Furthermore, it is related to the classic variant and possibly to thyroid extravasation.

Two-year analysis of efficacy and safety of deferasirox treatment for transfusional iron overload in sickle cell anemia patients.

The efficacy and safety of a 2-year treatment with deferasirox was evaluated in 31 patients with sickle cell anemia and transfusional iron overload. At 24 months, there were significant decreases from baseline in mean serum ferritin (from 2,344.6 to 1,986.3 µg/l; p = 0.040) and in mean liver iron concentration (from 13.0 ± 5.4 to 9.3 ± 5.7 mg Fe/g dry weight; p < 0.001). Myocardial T2* values were normal (>20 ms) in all patients at baseline and did not change significantly over the course of the study. However, there was a significant improvement from baseline in left ventricular ejection fraction at 24 months (62.2-64.6%; p = 0.02). Deferasirox was generally well tolerated with no progressive increases in serum creatinine or renal failure observed. These data confirm that deferasirox is effective in reducing body iron burden in patients with sickle cell anemia and transfusional iron overload.

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.

Mutação BRAF em pacientes idosos com carcinoma papilífero / BRAF mutations in the elderly with papillary carcinoma

Introdução: A mutação BRAF representa a alteração genética maiscomum do câncer de tireoide e esta foi a descoberta mais notávelneste campo de pesquisa, nos últimos anos; estão presentes em23-83% dos carcinomas papilíferos e são altamente específicaspara este tipo histológico. Método: Estudamos 22 pacientes,com mais de 65 anos, e diagnóstico de carcinoma papilíferosubmetidos a tireoidectomia, analisando a mutação BRAFV600E através de reação de PCR-RT realizada após a extraçãodo DNA dos blocos de parafina. Resultados: Conseguimosdetectar a ausência ou presença da mutação BRAF V600E em17 pacientes (77,3%). Entre estes 17 carcinomas papilíferos, seteapresentavam a mutação (41,2%). Demonstramos associaçãoestatística entre a presença desta mutação e a variante clássicado carcinoma papilífero, além de tendência de associação como extravasamento tireoideano. Discussão: Embora a mutaçãoBRAF seja a anormalidade genética mais comum no carcinomapapilífero, o seu significado prognóstico ainda não está bemestabelecido. Estudos contraditórios têm sido publicados,provavelmente pela heterogeneidade do carcinoma papilífero aoredor do mundo. Conclusão: a mutação BRAF, nos pacientesidosos com carcinoma papilífero tem frequência expressiva, eestá relacionada com sua variante clássica.

Correlação entre a ultrassonometria óssea do calcâneo e a densitometria em mulheres pós-menopausadas com fraturas por fragilidade óssea / Correlation between calcaneal bone ultrasound measurements and densitometry among postmenopausal women with fractures caused by bone fragility

OBJETIVO: Avaliar a correlação entre a ultrassonometria (US) do calcâneo e a densitometria (DEXA) em mulheres pós-menopausadas que já apresentavam uma fratura por fragilidade. MÉTODOS: Realizada coorte retrospectiva em 35 mulheres com fraturas osteoporóticas (punho ou coluna), deambulando, acima dos 40 anos, pós-menopausadas, sem tratamento prévio para osteoporose. Dessas, 16 com menos de 60 anos e 19 acima. Foram comparadas a Broadband Ultrasound Attenuation (BUA) e a Speed of Sound (SOS) com os sítios de DEXA (L1-L4, fêmur total, colo de fêmur e punhos), sendo utilizados dois valores de BUA diferentes como ponto de corte para osteoporose: BUA < 60dB/MHz e BUA < 64dB/MHz, e o SOS < 1.600m/s. O intervalo de confiança foi de 95 por cento. Os valores da DEXA e US foram lançados em um gráfico de dispersão, sendo possível, através de regressão linear, estabelecer correlações. Em seguida, a amostra foi estratificada segundo faixas etárias (até 60 e acima ou igual 60 anos). Desta forma, os valores foram novamente confrontados e correlacionados. RESULTADOS: A melhor correlação obtida entre DEXA e US foi entre o T score do punho e BUA < 64dB/MHz, com sensibilidade de 92 por cento e especificidade de 95 por cento. Foram obtidas melhores sensibilidades em todos os sítios da DEXA quando a US foi realizada em pacientes acima dos 60 anos. O SOS compatível com osteoporose é < 1.592,5m/s (sensibilidade de 89 por cento e especificidade de 85 por cento). CONCLUSÃO: US do calcâneo pode ser utilizada para rastreamento do risco de fratura por osteoporose quando utilizado ponto de corte BUA < 64dB/ MHz, principalmente em pacientes acima dos 60 anos.

OBJECTIVE: To assess the correlation between ultrasound (US) measurement on the calcaneus and bone densitometry (DEXA), among postmenopausal women who already presented fragility fractures. METHODS: 35 postmenopausal women over 40 years of age, with the ability to walk and presenting osteoporotic fractures of the wrist or spine, without previous treatment for osteoporosis, were analyzed in a retrospective cohort. Of these, 16 were under 60 and 19 were over 60. The broadband ultrasound attenuation (BUA) and speed of sound (SOS) were compared using DEXA (L1-L4, total femur, femoral neck and wrist). Two different values of BUA were used as cutoff points for osteoporosis: BUA < 60 dB/MHz and BUA < 64 dB/MHz (P < 0.05); and SOS < 1600 m/s. The confidence interval was 95 percent. The DEXA and US data were plotted on dispersion graphs and, through linear regression, it was possible to establish correlations. Following this, the sample was stratified according to age (up to 60 years and 60 years and over). Thus, the values were again compared and correlated. RESULTS: The best correlation obtained between DEXA and US was between the T-score of the wrist and BUA < 64 dB/ MHz, with 92 percent sensitivity and 95 percent specificity. Better sensitivity at all DEXA sites was obtained when US was performed on patients over 60 years of age. The SOS compatible with osteoporosis was < 1592.5 m/s (89 percent sensitivity and 85 percent specificity). CONCLUSION: US on the calcaneus can be used for screening the risk of osteoporosis fractures, using a cutoff of BUA < 64 dB/ MHz, especially among patients over 60 years of age.

New clinical score to diagnose nonalcoholic steatohepatitis in obese patients.

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is the most frequent disease associated with abnormal liver tests that is characterized by a wide spectrum of liver damage, ranging from simple macro vesicular steatosis to steatohepatitis (NASH), cirrhosis or liver carcinoma. Liver biopsy is the most precise test to differentiate NASH from other stages of NAFLD, but it is an invasive and expensive method. This study aimed to create a clinical laboratory score capable of identify individual with NASH in severely obese patients submitted to bariatric surgery. METHODS: The medical records from 66 patients submitted to gastroplasty were reviewed. Their chemistry profile, abdominal ultrasound (US) and liver biopsy done during the surgical procedure were analyzed. Patients were classified into 2 groups according to liver biopsy: Non-NASH group - those patients without NAFLD or with grade I, II or III steatosis; and NASH group - those with steatohepatitis or fibrosis. The t-test was used to compare each variable with normal distribution between NASH and Non-NASH groups. When comparing proportions of categorical variables, we used chi-square or z-test, where appropriate. A p-value < 0.05 was considered statistically significant. RESULTS: 83% of patients with obesity grades II or III showed NAFLD, and the majority was asymptomatic. Total Cholesterol (TC)≥200 mg/dL, alanine aminotransferase (ALT) ≥30, AST/ALT ratio (AAR)≤ 1, gammaglutaril-transferase (γGT)≥30 U/L and abdominal US, compatible with steatosis, showed association with NASH group. We proposed 2 scores: Complete score (TC, ALT, AAR, γGT and US) and the simplified score, where US was not included. The combination of biochemical and imaging results improved accuracy to 84.4% the recognition of NASH (sensitivity 70%, specificity 88.6%, NPV 91.2%, PPV 63. 6%). CONCLUSION: Alterations in TC, ALT, AAR, γGT and US are related to the most risk for NASH. The combination of biochemical and imaging results improved accuracy to 84.4% the recognition of NASH. Additionally, negative final scores exclude the presence of an advanced illness. Using this score, the severity of fatty liver infiltration would be predicted without the risks associated with hepatic biopsy.

CORRELATION BETWEEN CALCANEAL BONE ULTRASOUND MEASUREMENTS AND DENSITOMETRY AMONG POSTMENOPAUSAL WOMEN WITH FRACTURES CAUSED BY BONE FRAGILITY.

OBJECTIVE: To assess the correlation between ultrasound (US) measurement on the calcaneus and bone densitometry (DEXA), among postmenopausal women who already presented fragility fractures. METHODS: 35 postmenopausal women over 40 years of age, with the ability to walk and presenting osteoporotic fractures of the wrist or spine, without previous treatment for osteoporosis, were analyzed in a retrospective cohort. Of these, 16 were under 60 and 19 were over 60. The broadband ultrasound attenuation (BUA) and speed of sound (SOS) were compared using DEXA (L1-L4, total femur, femoral neck and wrist). Two different values of BUA were used as cutoff points for osteoporosis: BUA < 60 dB/MHz and BUA < 64 dB/MHz (P < 0.05); and SOS < 1600 m/s. The confidence interval was 95%. The DEXA and US data were plotted on dispersion graphs and, through linear regression, it was possible to establish correlations. Following this, the sample was stratified according to age (up to 60 years and 60 years and over). Thus, the values were again compared and correlated. RESULTS: The best correlation obtained between DEXA and US was between the T-score of the wrist and BUA < 64 dB/MHz, with 92% sensitivity and 95% specificity. Better sensitivity at all DEXA sites was obtained when US was performed on patients over 60 years of age. The SOS compatible with osteoporosis was < 1592.5 m/s (89% sensitivity and 85% specificity). CONCLUSION: US on the calcaneus can be used for screening the risk of osteoporosis fractures, using a cutoff of BUA < 64 dB/MHz, especially among patients over 60 years of age.

Myocilin mt.1 gene promoter single nucleotide polymorphism (-1000C>G) in Brazilian patients with primary open angle glaucoma.

BACKGROUND: The myocilin (MYOC) gene promoter polymorphism -1000C>G (MYOC mt.1) can be associated with faster progression of primary open angle glaucoma (POAG). The purpose of this study was to investigate the MYOC mt.1 in Brazilian patients with POAG and to evaluate its possible role on the phenotype and the severity of the disease. MATERIAL AND METHODS: One hundred sixty-seven POAG patients and 130 normal controls were enrolled. DNA samples were prepared and the MYOC mt.1 polymorphism was screened by real-time polymerase chain reaction (RT-PCR) in an Single-nucleotide polymorphism (SNP) assay. Frequencies of the MYOC mt.1 promoter polymorphism were determined for both groups and compared by Fisher's exact test and Chi-square test with Yate's correction. Intraocular pressure (IOP), cup-to-disc ratio (C/D), number of glaucoma medications, and number of glaucoma surgeries were compared between MYOC mt.1 carriers and non-carriers. RESULTS: MYOC mt.1 genotype frequencies did not differ between POAG and controls (P = 0.420); 14.6% of controls and 16.4% of POAG patients were MYOC mt.1 carriers (CG or GG). Frequencies of the G allele were similar between glaucomatous patients and controls (7.3% and 9.2%, respectively; P = 0.477). Among POAG patients, there were no differences in mean C/D ratio, IOP, number of glaucoma medications, and surgical procedures for IOP control between carries and non-carriers of the MYOC mt.1 promoter polymorphism (p>0.05). CONCLUSION: The G allele of the MYOC mt.1 promoter polymorphism was equally distributed among POAG patients and healthy subjects and it is possibly unrelated to the risk and severity of disease in the Brazilian population.

Miniaturization and globalization of clinical laboratory activities.

Clinical laboratories provide an invaluable service to millions of people around the world in the form of quality diagnostic care. Within the clinical laboratory industry the impetus for change has come from technological development (miniaturization, nanotechnology, and their collective effect on point-of-care testing; POCT) and the increasingly global nature of laboratory services. Potential technological gains in POCT include: the development of bio-sensors, microarrays, genetics and proteomics testing, and enhanced web connectivity. In globalization, prospective opportunities lie in: medical tourism, the migration of healthcare workers, cross-border delivery of testing, and the establishment of accredited laboratories in previously unexplored markets. Accompanying these impressive opportunities are equally imposing challenges. Difficulty transitioning from research to clinical use, poor infrastructure in developing countries, cultural differences and national barriers to global trade are only a few examples. Dealing with the issues presented by globalization and the impact of developing technology on POCT, and on the clinical laboratory services industry in general, will be a daunting task. Despite such concerns, with appropriate countermeasures it will be possible to address the challenges posed. Future laboratory success will be largely dependent on one's ability to adapt in this perpetually shifting landscape.

Coleta, transporte e armazenamento de amostras para diagnóstico molecular / Collection, transport and storage of samples for molecular diagnosis

A Sociedade Brasileira de Patologia Clínica/Medicina Laboratorial (SBPC/ML) lançou, em seu 44º Congresso Brasileiro de Patologia Clínica e Medicina Laboratorial, uma série de recomendações para minimizar os erros na fase pré-analítica, sendo este texto uma delas. Além do crescimento do mercado de testes moleculares, é frequente a percepção errônea de que esses testes são menos sujeitos a erros. Enfatizamos, neste documento, que certos cuidados na fase pré-analítica são extremamente importantes para garantir a confiabilidade de testes moleculares.

The Brazilian Society of Clinical Pathology/Laboratory Medicine (SBPC/ML) launched a series of recommendations on how to minimize errors on the pre-analytical testing phase during the 44th Brazilian Congress of Clinical Pathology and Laboratory Medicine. This is one of them. The molecular diagnostics market is growing fast and many people believe these tests are less error-prone. In this study we highlight that proper care during the pre-analytical phase is extremely important in order to ensure the reliability of molecular diagnostic tests.

Avaliação de métodos de concentração e detecção molecular de adenovírus em águas não tratadas - uma metanálise / Evaluación de métodos para concentración y detección molecular de adenovirus en agua no tratada: Un meta-análisis / Assessment of methods for the concentration and molecular detection of adenovirus in untreated water: A meta-analysis

Metodologias moleculares baseadas em PCR têm sido utilizadas para detectar adenovírus (AdVs) em amostras ambientais. É consenso entre os pesquisadores que estas metodologias oferecem vantagens sobre os métodos tradicionais de isolamento viral por cultura celular, sendo mais sensíveis, específicas e propiciando menor tempo de diagnóstico; contudo, há divergências em relação ao método de concentração viral a ser utilizado em amostras ambientais. Assim sendo, realizou-se uma metanálise com o intuito de responder aos questionamentos concernentes à eficácia do uso de três metodologias de concentração viral associadas à detecção molecular de AdVs em amostras de águas não tratadas, para a qual foram selecionados 33 estudos. Concluiu-se que: a) PCR não deve ser o método de escolha para detectar AdVs em amostras ambientais, devendo-se priorizar qPCR ou Nested-PCR; b) para detectar AdVs em amostras de rios ou lagos, a metodologia de escolha deve ser a associação entre ultracentrifugação e Nested-PCR; c) é aconselhável utilizar associação entre microfiltração em membrana, ultrafiltração e qPCR para detectar AdVs em amostras de esgotos tratados e não tratados. Estudos adicionais são necessários para avaliar os métodos que foram empregados em um único estudo e/ou com número escasso de amostras.

La detección de adenovirus (AdVs) en muestras ambientales se realiza por medio de metodologías moleculares basadas en PCR. Existe consenso entre los investigadores que estas metodologías ofrecen algunas ventajas en comparación con los métodos tradicionales de aislamiento de virus por medio de cultivos celulares; sin embargo, el método que se debe usar para concentrar los virus en muestras ambientales todavía es controversial. Por consiguiente, realizamos un meta-análisis dirigido a responder las preguntas respecto a la eficacia de tres métodos de concentración de virus asociados a la detección molecular de AdVs en muestras de agua no tratada, seleccionando 33 estudios. Concluimos que: a) el PCR no debe ser el método de elección para la detección de AdVs en muestras ambientales y que en vez debe usarse prioritariamente el método de qPCR o PCR-Anidada; b) para la detección de AdVs en muestras de agua tomadas de ríos o lagos, el método de elección debe ser una asociación de ultracentrifugación y PCR-Anidada; c) es aconsejable usar una asociación de membrana de microfiltración, ultrafiltración y qPCR para la detección de AdVs en muestras de aguas negras tratadas o no tratadas. Se necesitan más estudios para evaluar los métodos que se han usado en un solo estudio y/o con un número escaso de muestras.

Molecular methodologies based on PCR have been used for the detection of adenovirus (AdVs) in environmental samples. It is a consensus among researchers that these methodologies offer some advantages compared with traditional methods for the isolation of virus by cell culture, since they are more sensitive and specific and also require less processing time; however, the method to be used for virus concentration in environmental samples is still controversial. Consequently, we carried out a meta-analysis, aiming at responding the questions concerning the efficacy of three methods for virus concentration associated to the molecular detection of AdVs in untreated water samples, by selecting 33 studies. We concluded that: a) PCR should not be the method of choice for the detection of AdVs in environmental samples, and instead the use of qPCR or Nested-PCR should be prioritized; b) for the detection of AdVs in water samples collected in rivers or lakes, the method of choice should be an association of ultracentrifugation and Nested-PCR; c) it is advisable to use an association of microfiltration membrane, ultrafiltration, and qPCR for the detection of AdVs in treated and untreated sewage samples. Further studies are needed to evaluate the methods that have been used in only one study and/or with a limited number of samples.

Prevalence of Helicobacter spp in chronic cholecystitis and correlation with changes on the histological pattern of the gallbladder / Prevalência do Helicobacter spp na colecistite crônica calculosa e correlação com as alterações histológicas da vesícula biliar

PURPOSE: Establish the prevalence of Helicobacter spp in chronic cholecystitis and its correlation with the gallbladder's histological findings. METHODS: 100 patients were operated for chronic cholecystitis with cholecystolithiasis. In pathological examination of the gallbladder, were evaluated the presence of metaplasia, dysplasia, lymphoid follicles, anaplasia and tumors that might be related to the presence of Helicobacter plus the presence of the bacilli Giemsa by optical microscopy. From the DNA extracted from the gallbladder's bile, PCR was performed by using specific primers for the identification of Helicobacter spp with amplification of the 400bp segment of rRNA gene16S, with positive control DNA from Helicobacter pylori. All the cases negative for isolation of genetic material were excluded. The cases of PCR and Giemsa were used as negative control group. The histological findings were compared to the presence of bacilli and PCR data using a chi-square and Fisher's Exact test (CI = 95.0 percent, p <0.05). RESULTS: Of 68 patients, 42 (61.8 percent) were PCR for Helicobacter spp and 19 (27.9 percent) had Giemsa. There was no correlation between the two findings. The PCR for Helicobacter spp was not correlated to the histological findings. The presence of lymphoid follicles and metaplasia was related to the Giemsa (p = 0.025 and p= 0.039). CONCLUSION: There is high prevalence of Helicobacter spp in patients with chronic cholecystitis and cholecystolithiasis without be correlated with the histological patterns studied.

OBJETIVO: Estabelecer a prevalência do Helicobacter spp nos doentes com colecistopatia crônica calculosa e correlacioná-la com as alterações histológicas da vesícula biliar. MÉTODOS: Foram operados 100 doentes portadores de colecistite crônica calculosa. No anátomo-patológico foram avaliadas a presença de, metaplasias, displasias, folículos linfóides, anaplasias e tumores que pudessem se relacionar à presença do helicobacter e a presença de bacilos Giemsa à microscopia. A partir do DNA extraído da bile foi realizada PCR utilizando-se primers específicos para identificação de Helicobacter spp com amplificação de segmento de 400bp do gene16S rRNA, com controle positivo de DNA de Helicobacter Pylori. Os casos negativos para isolamento de material genético na bile foram excluídos. Os casos de PCR e Giemsa negativos foram utilizados como grupo controle. Os achados histológicos foram comparados ao Giemsa e à PCR utilizando-se Teste do Qui-Quadrado e Exato de Fisher (IC=95,0 por cento; p<0,05). RESULTADOS: De 68 doentes, 42(61,8 por cento) apresentaram PCR para Helicobacter spp e 19(27,9 por cento) Giemsa. Não houve correlação entre os dois achados. Não houve correlação entre alterações histológicas e a PCR (p=0,378). A presença de folículos linfóides e metaplasia estiveram relacionadas ao Giemsa (p=0,025 e 0,039). CONCLUSÃO: Há prevalência elevada de Helicobacter spp nos doentes com colecistite crônica calculosa sem haver correlação com os achados histológicos estudados.

Evaluation of weight loss and metabolic profile of obese patients with type 2 diabetes mellitus (dm2) versus non-dm2 subjects, one and three years after bariatric surgery / Avaliação da perda ponderal e perfil metabólico pós-cirurgia bariátrica em pacientes obesos portadores de diabetes mellitus tipo 2 versus não diabéticos

BACKGROUND: There is a debate over results obtained from type 2 diabetes mellitus (DM2) obese patients and non-DM2 patients, in reference to metabolic control and ponderal loss, after bariatric surgery. AIM: To evaluate weight loss and metabolic profile of obese patients with DM2 versus non-DM2 subjects, one and three years after bariatric surgery. METHODS: Data from 38 non-DM2 patients and 44 DM2 patients submitted to Roux-en-Y gastric-bypass were analysed retrospectively. For the pre-operatory, first and third year of post-operatory, were compared: weight, body mass index (BMI), fasting glucose (FG), high density lipoprotein (HDL) and triglycerides (TG). RESULTS: Preoperatively, both groups were statistically equivalent in regards to weight, BMI (P = 0.90) and HDL (P = 0.73). This was not the case when TG (P = 0.043) and FG (P<0.01) were analyzed. In PO1, both DM2 and non-DM2 groups showed a reduction in weight, BMI and TG, just as FG in the DM2 group (P < 0.05). HDL increased (P < 0.05) in PO1 in both groups. In the following period, between PO1 and PO3, only TG continued to decrease in the non-DM2 group (P = 0.039), while the other variables did not change. In the DM2 group mean A1c in PO3 was 6.2

RACIONAL: Tem havido debate sobre os resultados obtidos da cirurgia bariátrica nos obesos diabéticos versus não diabéticos, no quesito controle metabólico e perda ponderal. OBJETIVO: Avaliar a perda ponderal e perfil metabólico dos obesos diabéticos e não diabéticos após um e três anos da cirurgia bariátrica. MÉTODO: Análise retrospectiva dos prontuários de 44 diabéticos tipo 2 (DM2) e 38 não-DM2 submetidos à gastroplastia com derivação em Y-de-Roux. Dados como peso, índice de massa corpórea (IMC), glicemia de jejum (GJ), lipoproteína de alta densidade (HDL) e triglicérides (TG) foram vistos no pré-operatório, primeiro (PO1) e terceiro ano (PO3) após a operação. RESULTADOS: No pré-operatório, ambos os grupos foram estatisticamente equivalentes em peso, IMC (P=0,90) e HDL (P=0.73). Não se verificou o mesmo quando TG (P=0.043) e GJ (P < 0.01) foram analisados. No PO1, ambos DM2 e não-DM2 mostraram redução no peso, IMC e TG, assim como GJ no grupo DM2 (P < 0.05). HDL aumentou (P < 0.05) no PO1 em ambos os grupos. No período seguinte, entre PO1 e PO3, somente TG continuou a cair nos não-DM2 diabéticos (P=0,039), enquanto as outras variáveis não mudaram. No grupo DM2, a média da A1c no PO3 foi 6,2

+- 0,75 (P=0,027). Compararam-se os dados do pós-operatório de ambos os grupos. A variação de HDL e TG entre os grupos não foi diferente no PO1 e entre PO1 e PO3. Peso no PO1 e PO3, assim como IMC no PO1 e PO3, não foram estatisticamente diferentes. CONCLUSÃO: No PO1, perda de peso e melhora metabólica foi vista em ambos os grupos. Este dado foi sustentado no PO3, sem reganho de peso ou alteração lipídica significativa. A A1c encontrada no pós-operatório mostrou bom controle do diabetes pela operação. Houve tendência à menor perda ponderal no grupo DM2 (P=0,053).

+- 0.75 (P = 0.027). It was compared both group's post-operative data. HDL's and TG's variation between groups did not differ in PO1 or between PO1 and PO3. Weight in PO1 and PO3, just as BMI in PO1 and PO3, were not significantly different either. CONCLUSION: In PO1, weight loss and metabolic improvement was seen in both groups. This was sustained in PO3, with no significant weight regain or lipid/FG change. A1c found suggests a reasonable control of DM2 surgery. A trend towards a less intense weight loss could be noticed in the DM2 group (P = 0.053).

Prevalence of Helicobacter spp in chronic cholecystitis and correlation with changes on the histological pattern of the gallbladder.

PURPOSE: Establish the prevalence of Helicobacter spp in chronic cholecystitis and its correlation with the gallbladder's histological findings. METHODS: 100 patients were operated for chronic cholecystitis with cholecystolithiasis. In pathological examination of the gallbladder, were evaluated the presence of metaplasia, dysplasia, lymphoid follicles, anaplasia and tumors that might be related to the presence of Helicobacter plus the presence of the bacilli Giemsa(+) by optical microscopy. From the DNA extracted from the gallbladder's bile, PCR was performed by using specific primers for the identification of Helicobacter spp with amplification of the 400bp segment of rRNA gene16S, with positive control DNA from Helicobacter pylori. All the cases negative for isolation of genetic material were excluded. The cases of PCRTheta and GiemsaTheta were used as negative control group. The histological findings were compared to the presence of bacilli and PCR data using a chi-square and Fisher's Exact test (CI = 95.0%, p <0.05). RESULTS: Of 68 patients, 42 (61.8%) were PCR(+) for Helicobacter spp and 19 (27.9%) had Giemsa(+). There was no correlation between the two findings. The PCR(+) for Helicobacter spp was not correlated to the histological findings. The presence of lymphoid follicles and metaplasia was related to the Giemsa(+) (p = 0.025 and p= 0.039). CONCLUSION: There is high prevalence of Helicobacter spp in patients with chronic cholecystitis and cholecystolithiasis without be correlated with the histological patterns studied.