Understanding the relation between Zika virus infection during pregnancy and adverse fetal, infant and child outcomes: a protocol for a systematic review and individual participant data meta-analysis of longitudinal studies of pregnant women and their infants and children

IntroductionZika virus (ZIKV) infection during pregnancy is a known cause of microcephaly and other congenital and developmental anomalies. In the absence of a ZIKV vaccine or prophylactics, principal investigators (PIs) and international leaders in ZIKV research have formed the ZIKV Individual Participant Data (IPD) Consortium to identify, collect and synthesise IPD from longitudinal studies of pregnant women that measure ZIKV infection during pregnancy and fetal, infant or child outcomes.Methods and analysisWe will identify eligible studies through the ZIKV IPD Consortium membership and a systematic review and invite study PIs to participate in the IPD meta-analysis (IPD-MA). We will use the combined dataset to estimate the relative and absolute risk of congenital Zika syndrome (CZS), including microcephaly and late symptomatic congenital infections; identify and explore sources of heterogeneity in those estimates and develop and validate a risk prediction model to identify the pregnancies at the highest risk of CZS or adverse developmental outcomes. The variable accuracy of diagnostic assays and differences in exposure and outcome definitions means that included studies will have a higher level of systematic variability, a component of measurement error, than an IPD-MA of studies of an established pathogen. We will use expert testimony, existing internal and external diagnostic accuracy validation studies and laboratory external quality assessments to inform the distribution of measurement error in our models. We will apply both Bayesian and frequentist methods to directly account for these and other sources of uncertainty.Ethics and disseminationThe IPD-MA was deemed exempt from ethical review. We will convene a group of patient advocates to evaluate the ethical implications and utility of the risk stratification tool. Findings from these analyses will be shared via national and international conferences and through publication in open access, peer-reviewed journals.Trial registration numberPROSPERO International prospective register of systematic reviews (CRD42017068915).</jats:sec

Monitoramento da reativação do BK poliomavírus em pacientes transplantados renais no Hospital Universitário Antonio Pedro

Introdução: A nefropatia associada ao BK poliomavírus acomete cerca de 10 % dos pacientes transplantados renais, com perda de função do enxerto em até 50 % dos casos da doença. Muitos autores recomendam a PCR no plasma/urina como estratégia de monitoramento dos pacientes no pós-transplante, visando diagnóstico precoce da reativação do poliomavírus. Hipótese científica: O monitoramento citológico urinário dos pacientes transplantados renais por meio da pesquisa de decoy cells é importante para o acompanhamento clínico póstransplante, objetivando identificação precoce da reativação viral e seleção dos pacientes com maior risco de desenvolvimento da nefropatia associada ao poliomavírus. Objetivo geral: Avaliar a eficácia do monitoramento plasmático e urinário com fins de detecção do DNA do BKV e pesquisa urinária de decoy cells em uma coorte de pacientes transplantados renais, visando identificar os indivíduos com maior risco de desenvolvimento da nefropatia. Objetivos específicos: Pesquisar decoy cells em urina e DNA do BK poliomavírus em sangue e urina do grupo de pacientes; analisar as amostras de biopsia do enxerto renal realizadas no período do estudo; avaliar o curso clínico dos pacientes, correlacionando-o com os resultados do monitoramento molecular e citológico. Material e métodos: Pacientes transplantados renais foram acompanhados com coletas quinzenais de urina até o terceiro mês, uma coleta ao sexto mês e uma após um ano de transplante. A urina foi submetida a pesquisa de decoy cells e PCR quantitativa para BKV. Amostras de sangue foram coletadas ao final do segundo e terceiro mês pós-transplante para realização da PCR. Foi realizada pesquisa do poliomavírus por imuno-histoquímica nas amostras de biopsia do enxerto renal. Amostras urinárias contendo decoy cells foram submetidas a exame ultraestrutural, imunocitoquímico e por microscopia de contraste de fase. Resultados: O estudo envolveu 29 pacientes. A pesquisa viral urinária foi realizada em 19 pacientes e mostrou positividade em 16 (84 %), sendo que em 13 (81 %) detectada nos primeiros três meses. A pesquisa de decoy cells foi realizada em 25 pacientes, sendo positiva em seis (24 %) dos quais quatro (67 %) apresentaram positividade no terceiro mês pós-transplante. A pesquisa viral plasmática foi realizada em 22 indivíduos sendo positiva em seis (27 %), todos ao final do terceiro mês pós-transplante. A positividade para decoy cells associou-se ao histórico prévio de transfusão sanguínea do receptor (p=0,012). A análise temporal das três técnicas de monitoramento revelou poder de detecção precoce de positividade na seguinte ordem: DNA viral urinário, decoy cells e DNA viral plasmático. Apenas um caso de nefropatia foi diagnosticado ao final do primeiro ano do transplante. Este paciente eliminou, por 45 semanas, numerosas decoy cells isoladas e constituindo cilindros, além de apresentar numerosas partículas virais intra e extracelulares sob exame ultraestrutural da urina. Conclusão: O monitoramento precoce dos pacientes no pós-transplante renal, por meio da pesquisa de decoy cells, foi efetivo na detecção da reativação do poliomavírus. A positividade para decoy cells mostrou-se preditora de nefropatia pel o BK poliomavírus em caso de eliminação sustentada (≥ 6 semanas) associada a fundo citológico produtivo, mesmo na ausência de disfunção clínica do enxerto. O monitoramento, ao ser desenvolvido contínua e periodicamente pelo período mínimo de um ano após o transplante, revelou-se estratégia promissora para seleção e tratamento tempestivo de pacientes com maior risco de nefropatia associada ao poliomavírusIntroduction: BK polyomavirus-associated nephropathy occurs in about 10 % of kidney transplant patients, leading to loss of graft in up to 50% of the cases of nephropathy. Many authors recommend plasma/urine PCR for BKV-DNA as a strategy to monitoring posttransplant patients for early detection of polyomavirus reactivation. Scientific hypothesis: Urinary monitoring in renal transplant patients through decoy cells screening is an important strategy during the post-transplant period, allowing early detection of polyomavirus reactivation and identification of patients with a higher risk of developing BKV-associated nephropathy. General objective: Evaluate the importance of monitoring urine and plasma to detect BKVDNA and screening urinary decoy cells in a group of renal transplant recipients, aiming to identify patients at higher risk of developing nephropathy. Specific objectives: Search for decoy cells in urine samples and BKV-DNA in blood and urine samples obtained from the group of patients; analyze the graft samples obtained during the study; evaluate the patient follow-up and correlate it with the monitoring results. Material and methods: Renal transplant patients were monitored with biweekly urine collection up to the third month post-transplant, one collection at sixth months and one after a year. The urine samples were submitted to cytological examination to search for decoy cells and to quantitative PCR assays to identify BKV-DNA. Blood samples were collected at the end of the second and third month posttransplant for PCR testing for BKV-DNA. Immunohistochemistry analysis for polyomavirus were performed in the graft samples. In addition, urine samples presenting decoy cells were submitted to ultrastructural analyses, immunocytochemistry for polyomavirus and phasecontrast microscopy studies. Results: The study included 29 patients. PCR urinary test was performed in 19 patients, and they were positive in 16 (84 %) of the cases. Thirteen (81 %) of these cases were detected during the first three months. Decoy cells analyses were performed in 25 patients, with positive results in six (24 %). Four of them (67 %) were detected at the third month. BKV-DNA blood tests were performed in 22 patients, with positive results in six patients (27 %), at end of third post-transplant month. Decoy cells positivity was associated to previous history of blood transfusion for the receptor (p=0,012). A timeline analysis of the three monitoring techniques revealed the following order in terms of early detection power: urinary BKV-DNA, decoy cells, and plasma BKV-DNA. Just one case of BKV-associated nephropathy was diagnosed at end of the first year post-transplant. This patient presented shedding of a huge number of decoy cells for 45 weeks, as single cells or arranged in casts, and presented abundant intra and extracellular viral particles, detected through ultrastructural analysis of urine samples. Conclusion: Early post-transplant monitoring by urinary decoy cells analysis was effective in BKV reactivation detection. Decoy cells positivity was predictive for BKVassociated nephropathy in the case of sustained shedding (≥ 6 weeks) related to dirty cytological background, even in the absence of clinical graft dysfunction. Monitoring, when carried out continuously and periodically for a minimum of one year after transplantation, was shown to be a promising strategy for the selection and timely treatment of patients at higher risk of developing BKV-associated nephropathy131f

Condyloma acuminatum: its histopathological pattern

Condyloma acuminatum is one of the clinical manifestations of papillomavirus infection. The classical histopathological features are already known and do not constitute a diagnostic problem. Clinically, it has been classified into growth or proliferative, full-expression, and regressive or persistent phases, with the histopathological aspects of these distinct phases being well documented in equine cutaneous papillomas. We have designed a protocol of histopathological analysis in order to investigate the possibility of identifying the evolutional phases in human condylomata acuminata. Sixty condylomata acuminata from the files of the Department of Pathology, Universidade Federal Fluminense, were studied regarding koilocytosis, paraceratosis, acantosis, basal cell hyperplasia and mono-nuclear cell infiltrate. After an individual analysis and comparison of the cases, the main differential aspects of condyloma acuminatum were: koilocytosis, transepithelial lymphocytic infiltrate and basal cell hyperplasia. Thus, condylomatous lesions can be histopathologically differentiated in three major patterns: proliferative, viral replication activity and regressive

Hemophagocytic lymphohistiocytosis, a rare condition in renal transplant - a case report

Abstract Hemophagocytic lymphohistiocytosis (HLH) is an uncommon and life-threating condition characterized by major immune activation and massive cytokine production by mononuclear inflammatory cells, due to defects in cytotoxic lymphocyte function. It is even more unusual in renal transplant recipients, in which it is often associated with uncontrolled infection. The mortality is high in HLH and differential diagnosis with sepsis is a challenge. The approach and management depend on the underlying trigger and comorbidities. We report a case of a 50-year-old renal transplant female admitted with fever and malaise 3 months post-transplant and presenting anemia, fever, hypertriglyceridemia, high levels of serum ferritin, and positive CMV antigenemia. Urine was positive for decoy cells and BKV-DNA. Graft biopsy showed CMV nephritis. Both blood and urine cultures where positive for E. coli. Hemophagocytosis was confirmed by bone marrow aspiration. Immunosuppression was reduced, and the patient received high-dose intravenous immunoglobulin and dexamethasone, with complete response after 3 weeks. We highlight the importance of early diagnosis and proper management of a rare and serious condition in a renal transplant patient, which can allow a favorable clinical course and improve survival rate

BK polyomavirus nephropathy in two kidney transplant patients with distinct diagnostic strategies for BK virus and similar clinical outcomes: two case reports

Abstract Background BK polyomavirus-associated nephropathy is an important cause of post-transplantation renal failure. We present two cases of BK polyomavirus-associated nephropathy who were submitted to contrasting strategies of clinical follow-up to BK polyomavirus reactivation, but progressed to a similar final outcome. Case presentation Case 1 is a 37-year-old white man whose graft had never presented a good glomerular filtration rate function, with episodes of tacrolimus nephrotoxicity, and no urinary monitoring for BK polyomavirus; stage B BK polyomavirus-associated nephropathy was diagnosed by biopsy at 14 months post-transplant. Despite clinical treatment (dosage decrease and immunosuppressive drug change), he progressed to stage C BK polyomavirus-associated nephropathy and loss of graft function 30 months post-transplant. Case 2 is a 49-year-old mulatto man in his second renal transplantation who was submitted to cytological urinary monitoring for BK polyomavirus; he presented early, persistent, and massive urinary decoy cell shedding and concomitant tacrolimus nephrotoxicity. Even with decreasing immunosuppression, he developed BK polyomavirus-associated nephropathy 1-year post-transplant. Loss of graft function occurred 15 months post-transplant. Conclusions Cytological urinary monitoring was an efficient strategy for monitoring BK virus reactivation. Decoy cell shedding may be related to BK polyomavirus-associated nephropathy when extensive and persistent. The presence of associated tacrolimus nephrotoxicity may be a confounding factor for the clinical diagnosis of BK polyomavirus-associated nephropathy

Nanographene oxide-methylene blue as phototherapies platform for breast tumor ablation and metastasis prevention in a syngeneic orthotopic murine model

Abstract Background In the photodynamic therapy (PDT), the photosensitizer absorbs light and transfers the energy of the excited state to the oxygen in the cell environment producing reactive oxygen species (ROS), that in its turn, may cause cell damage. In the photothermal therapy (PTT), light also is responsible for activating the photothermal agent, which converts the absorbed energy in heat. Graphene oxide is a carbon-based material that presents photothermal activity. Its physical properties allow the association with the photosensitizer methylene blue and consequently the production of ROS when submitted to light irradiation. Therefore, the association between nanographene oxide and methylene blue could represent a strategy to enhance therapeutic actions. In this work, we report the nanographene oxide-methylene blue platform (NanoGO-MB) used to promote tumor ablation in combination with photodynamic and photothermal therapies against a syngeneic orthotopic murine breast cancer model. Results In vitro, NanoGO-MB presented 50% of the reactive oxygen species production compared to the free MB after LED light irradiation, and a temperature increase of ~ 40 °C followed by laser irradiation. On cells, the ROS production by the nanoplatform displayed higher values in tumor than normal cells. In vivo assays demonstrated a synergistic effect obtained by the combined PDT/PTT therapies using NanoGO-MB, which promoted complete tumor ablation in 5/5 animals. Up to 30 days after the last treatment, there was no tumor regrowth compared with only PDT or PTT groups, which displayed tumoral bioluminescence 63-fold higher than the combined treatment group. Histological studies confirmed that the combined therapies were able to prevent tumor regrowth and liver, lung and spleen metastasis. In addition, low systemic toxicity was observed in pathologic examinations of liver, spleen, lungs, and kidneys. Conclusions The treatment with combined PDT/PTT therapies using NanoGO-MB induced more toxicity on breast carcinoma cells than on normal cells. In vivo, the combined therapies promoted complete tumor ablation and metastasis prevention while only PDT or PTT were unable to stop tumor development. The results show the potential of NanoGO-MB in combination with the phototherapies in the treatment of the breast cancer and metastasis prevention