A large-scale retrospective study enabled deep-learning based pathological assessment of frozen procurement kidney biopsies to predict graft loss and guide organ utilization.

Lesion scores on procurement donor biopsies are commonly used to guide organ utilization for deceased-donor kidneys. However, frozen sections present challenges for histological scoring, leading to inter- and intra-observer variability and inappropriate discard. Therefore, we constructed deep-learning based models to recognize kidney tissue compartments in hematoxylin & eosin-stained sections from procurement needle biopsies performed nationwide in years 2011-2020. To do this, we extracted whole-slide abnormality features from 2431 kidneys and correlated with pathologists' scores and transplant outcomes. A Kidney Donor Quality Score (KDQS) was derived and used in combination with recipient demographic and peri-transplant characteristics to predict graft loss or assist organ utilization. The performance on wedge biopsies was additionally evaluated. Our model identified 96% and 91% of normal/sclerotic glomeruli respectively; 94% of arteries/arterial intimal fibrosis; 90% of tubules. Whole-slide features of Sclerotic Glomeruli (GS)%, Arterial Intimal Fibrosis (AIF)%, and Interstitial Space Abnormality (ISA)% demonstrated strong correlations with corresponding pathologists' scores of all 2431 kidneys, but had superior associations with post-transplant estimated glomerular filtration rates in 2033 and graft loss in 1560 kidneys. The combination of KDQS and other factors predicted one- and four-year graft loss in a discovery set of 520 kidneys and a validation set of 1040 kidneys. By using the composite KDQS of 398 discarded kidneys due to "biopsy findings", we suggest that if transplanted, 110 discarded kidneys could have had similar survival to that of other transplanted kidneys. Thus, our composite KDQS and survival prediction models may facilitate risk stratification and organ utilization while potentially reducing unnecessary organ discard.

A Phase I Trial of VEGF-A Inhibition Combined with PD-L1 Blockade for Recurrent Glioblastoma.

Purpose: The treatment of glioblastoma (GBM) poses challenges. The use of immune checkpoint inhibition (ICI) has been disappointing as GBM is characterized by low mutational burden and low T-cell infiltration. The combination of ICI with other treatment modalities may improve efficacy. Patient and Methods: Patients with recurrent GBM were treated with avelumab, a human IgG1 antibody directed against PD-L1 (part A), or avelumab within a week after laser interstitial thermal therapy (LITT) and continuation of avelumab (part B). Bevacizumab was allowed to be combined with ICI to spare steroid use. The primary objective was to characterize the tolerability and safety of the regimens. The secondary objectives included overall survival, progression-free survival (PFS), signatures of plasma analytes, and immune cells. Results: A total of 12 patients (median age 64; range, 37-73) enrolled, five in part A and seven in part B. Two serious adverse events occurred in the same patient, LITT treated, not leading to death. The median survival from enrollment was 13 months [95% confidence interval (CI), 4-16 months] with no differences for part A or B. The median PFS was 3 months (95% CI, 1.5-4.5 months). The decrease in MICA/MICB, γδT cells, and CD4+ T cell EMRA correlated with prolonged survival. Conclusions: Avelumab was generally well tolerated. Adding bevacizumab to ICI may be beneficial by lowering cytokine and immune cell expression. The development of this combinatorial treatment warrants further investigation. Exploring the modulation of adaptive and innate immune cells and plasma analytes as biomarker signatures may instruct future studies in this dismal refractory disease. Significance: Our phase I of PD-L1 inhibition combined with LITT and using bevacizumab to spare steroids had a good safety profile for recurrent GBM. Developing combinatory treatment may help outcomes. In addition, we found significant immune modulation of cytokines and immune cells by bevacizumab, which may enhance the effect of ICI.

NKG2A and HLA-E define an alternative immune checkpoint axis in bladder cancer.

Programmed cell death protein 1 (PD-1)/programmed death-ligand 1 (PD-L1)-blockade immunotherapies have limited efficacy in the treatment of bladder cancer. Here, we show that NKG2A associates with improved survival and responsiveness to PD-L1 blockade immunotherapy in bladder tumors that have high abundance of CD8+ T cells. In bladder tumors, NKG2A is acquired on CD8+ T cells later than PD-1 as well as other well-established immune checkpoints. NKG2A+ PD-1+ CD8+ T cells diverge from classically defined exhausted T cells through their ability to react to human leukocyte antigen (HLA) class I-deficient tumors using T cell receptor (TCR)-independent innate-like mechanisms. HLA-ABC expression by bladder tumors is progressively diminished as disease progresses, framing the importance of targeting TCR-independent anti-tumor functions. Notably, NKG2A+ CD8+ T cells are inhibited when HLA-E is expressed by tumors and partly restored upon NKG2A blockade in an HLA-E-dependent manner. Overall, our study provides a framework for subsequent clinical trials combining NKG2A blockade with other T cell-targeted immunotherapies, where tumors express higher levels of HLA-E.

1-Deoxysphinganine initiates adaptive responses to serine and glycine starvation in cancer cells via proteolysis of sphingosine kinase.

Cancer cells may depend on exogenous serine, depletion of which results in slower growth and activation of adaptive metabolic changes. We previously demonstrated that serine and glycine (SG) deprivation causes loss of sphingosine kinase 1 (SK1) in cancer cells, thereby increasing the levels of its lipid substrate, sphingosine (Sph), which mediates several adaptive biological responses. However, the signaling molecules regulating SK1 and Sph levels in response to SG deprivation have yet to be defined. Here, we identify 1-deoxysphinganine (dSA), a noncanonical sphingoid base generated in the absence of serine from the alternative condensation of alanine and palmitoyl CoA by serine palmitoyl transferase, as a proximal mediator of SG deprivation in SK1 loss and Sph level elevation upon SG deprivation in cancer cells. SG starvation increased dSA levels in vitro and in vivo and in turn induced SK1 degradation through a serine palmitoyl transferase-dependent mechanism, thereby increasing Sph levels. Addition of exogenous dSA caused a moderate increase in intracellular reactive oxygen species, which in turn decreased pyruvate kinase PKM2 activity while increasing phosphoglycerate dehydrogenase levels, and thereby promoted serine synthesis. We further showed that increased dSA induces the adaptive cellular and metabolic functions in the response of cells to decreased availability of serine likely by increasing Sph levels. Thus, we conclude that dSA functions as an initial sensor of serine loss, SK1 functions as its direct target, and Sph functions as a downstream effector of cellular and metabolic adaptations. These studies define a previously unrecognized "physiological" nontoxic function for dSA.

Women's obstetric history and midtrimester cervical length measurements by 2d/3d and doppler ultrasound / História obstétrica e medida do comprimento do colo uterino de mulheres no segundo trimestre gestacional por ultrassonografia bi/tridimensional e Doppler

Abstract Objective The aim of the present study was to compare the obstetric history and both two- and tri-dimensional ultrasound parameters according to different cervical lengths. Methods The present cross-sectional study analyzed 248 midtrimester pregnant women according to cervical length and compared the data with the obstetric history and 2D/3D ultrasound parameters. Patients were divided into 3 groups according to cervical length: The Short Cervix group for cervical lengths ≥ 15mm and< 25mm(n= 68), the Very Short Cervix group for cervical lengths< 15mm (n = 18) and the Control group, composed of 162 pregnant women with uterine cervical lengths ≥ 25mm. Results When analyzing the obstetric history of only non-nulliparous patients, a significant association between the presence of a short cervix in the current pregnancy and at least one previous preterm birth was reported (p = 0.021). Cervical length and volume were positively correlated (Pearson coefficient = 0.587, p < 0.0001). The flow index (FI) parameter of cervical vascularization was significantly different between the Control and Very Short Cervix groups. However, after linear regression, in the presence of volume information, we found no association between the groups and FI. Uterine artery Doppler was also not related to cervical shortening. Conclusion The present study showed a significant association between the presence of a short cervix in the current pregnancy and at least one previous preterm birth. None of the vascularization indexes correlate with cervical length as an independent parameter. Uterine artery Doppler findings do not correlate with cervical length.

Resumo Objetivo O objetivo do presente estudo foi comparar a história obstétrica e os parâmetros bi- e tridimensionais ultrassonográficos de acordo com os diferentes comprimentos cervicais. Métodos O presente estudo transversal analisou 248 gestantes no segundo trimestre de acordo com o comprimento cervical e comparou os dados com a história obstétrica e os parâmetros ultrassonográficos 2D/3D. As pacientes foram divididas em 3 grupos de acordo com o comprimento do colo uterino: grupo Colo Curto para comprimentos cervicais ≥ 15mm e < 25mm (n = 68), grupo Colo Muito Curto para comprimentos cervicais < 15mm (n = 18) e grupo Controle, composto por 162 gestantes com comprimento cervical uterino ≥ 25 mm. Resultados Ao analisar a história obstétrica apenas de pacientes não nulíparas, foi relatadauma associação significativa entre a presença de colo uterino curto na gravidez atual e pelo menos um episódio de parto prematuro anterior (p = 0,021). Comprimento e volume do colo uterino foram correlacionados positivamente (coeficiente de Pearson = 0,587, p < 0,0001). O parâmetro índice de fluxo (IF) da vascularização cervical foi significativamente diferente entre os grupos Controle e Colo Muito Curto. Entretanto, após regressão linear, na presença de informações de volume, não encontramos associação entre os grupos e o parâmetro IF. Também não foi encontrada relação entre o Doppler da artéria uterina e o encurtamento cervical. Conclusão O presente estudo mostrou uma associação significativa entre a presença de colo uterino curto na gravidez atual e pelo menos um episódio de parto prematuro anterior. Nenhum dos índices de vascularização se correlaciona com o comprimento cervical como parâmetro independente, assim como o Doppler da artéria uterina também não está relacionado ao comprimento do colo uterino.

Women's Obstetric History and Midtrimester Cervical Length Measurements by 2D/3D and Doppler Ultrasound.

OBJECTIVE: The aim of the present study was to compare the obstetric history and both two- and tri-dimensional ultrasound parameters according to different cervical lengths. METHODS: The present cross-sectional study analyzed 248 midtrimester pregnant women according to cervical length and compared the data with the obstetric history and 2D/3D ultrasound parameters. Patients were divided into 3 groups according to cervical length: The Short Cervix group for cervical lengths ≥ 15 mm and < 25 mm (n = 68), the Very Short Cervix group for cervical lengths < 15 mm (n = 18) and the Control group, composed of 162 pregnant women with uterine cervical lengths ≥ 25 mm. RESULTS: When analyzing the obstetric history of only non-nulliparous patients, a significant association between the presence of a short cervix in the current pregnancy and at least one previous preterm birth was reported (p = 0.021). Cervical length and volume were positively correlated (Pearson coefficient = 0.587, p < 0.0001). The flow index (FI) parameter of cervical vascularization was significantly different between the Control and Very Short Cervix groups. However, after linear regression, in the presence of volume information, we found no association between the groups and FI. Uterine artery Doppler was also not related to cervical shortening. CONCLUSION: The present study showed a significant association between the presence of a short cervix in the current pregnancy and at least one previous preterm birth. None of the vascularization indexes correlate with cervical length as an independent parameter. Uterine artery Doppler findings do not correlate with cervical length.

OBJETIVO: O objetivo do presente estudo foi comparar a história obstétrica e os parâmetros bi- e tridimensionais ultrassonográficos de acordo com os diferentes comprimentos cervicais. MéTODOS: O presente estudo transversal analisou 248 gestantes no segundo trimestre de acordo com o comprimento cervical e comparou os dados com a história obstétrica e os parâmetros ultrassonográficos 2D/3D. As pacientes foram divididas em 3 grupos de acordo com o comprimento do colo uterino: grupo Colo Curto para comprimentos cervicais ≥ 15 mm e < 25 mm (n = 68), grupo Colo Muito Curto para comprimentos cervicais < 15 mm (n = 18) e grupo Controle, composto por 162 gestantes com comprimento cervical uterino ≥ 25 mm. RESULTADOS: Ao analisar a história obstétrica apenas de pacientes não nulíparas, foi relatada uma associação significativa entre a presença de colo uterino curto na gravidez atual e pelo menos um episódio de parto prematuro anterior (p = 0,021). Comprimento e volume do colo uterino foram correlacionados positivamente (coeficiente de Pearson = 0,587, p < 0,0001). O parâmetro índice de fluxo (IF) da vascularização cervical foi significativamente diferente entre os grupos Controle e Colo Muito Curto. Entretanto, após regressão linear, na presença de informações de volume, não encontramos associação entre os grupos e o parâmetro IF. Também não foi encontrada relação entre o Doppler da artéria uterina e o encurtamento cervical. CONCLUSãO: O presente estudo mostrou uma associação significativa entre a presença de colo uterino curto na gravidez atual e pelo menos um episódio de parto prematuro anterior. Nenhum dos índices de vascularização se correlaciona com o comprimento cervical como parâmetro independente, assim como o Doppler da artéria uterina também não está relacionado ao comprimento do colo uterino.

Emergency cerclage: gestational and neonatal outcomes

SUMMARY BACKGROUND: The gestational and neonatal outcomes of women with early cervical dilatation undergoing emergency cerclage were evaluated and compared with women treated with expectant management and bed rest. METHODS: Retrospective analysis of pregnant women admitted between 2001 and 2017 with a diagnosis of early cervical dilatation and/or bulging membranes. Patients with a singleton pregnancy of a fetus without malformations, between 16 and 25 weeks and 6 days, with cervical dilatation of 1 to 3 cm were included; patients who delivered or miscarried within 2 days after admission were excluded. RESULTS: The study enrolled 30 patients: 19 in the cerclage group and 11 in the rest group. There was a significant difference, with the cerclage group showing better results concerning gestational age at delivery (28.7 vs. 23.3 weeks; p=0.031) and latency between hospital admission and delivery (48.6 vs. 16 days; p=0.016). The fetal death rate was lower in the cerclage group (5.3% vs. 54.5%, p=0.004). Considering gestational age at delivery of live newborns, no difference was observed between the cerclage and rest groups (29.13 vs. 27.4 weeks; p=0.857). CONCLUSIONS: Emergency cerclage was associated with longer latency, a significant impact on gestational age at delivery and reduction in the fetal death rate.

RESUMO OBJETIVO: Os resultados gestacionais e neonatais de mulheres com cervicodilatação precoce submetidas à cerclagem de emergência foram avaliados e comparados com mulheres tratadas com manejo expectante com repouso no leito. MÉTODOS: Análise retrospectiva de gestantes admitidas entre 2001 e 2017 com diagnóstico de cervicodilatação precoce e/ou membranas protrusas. Foram incluídas pacientes com gestação única de feto sem malformações, entre 16 semanas e 25 semanas e 6 dias, com dilatação cervical de 1 a 3 cm; as pacientes que tiveram parto ou aborto dentro de 2 dias após admissão foram excluídas. RESULTADOS: O estudo envolveu 30 pacientes: 19 no grupo cerclagem e 11 no grupo repouso. Houve diferença significativa, com o grupo cerclagem apresentando melhores resultados em relação à idade gestacional no parto (28,7 vs. 23,3 semanas; p=0,031) e à latência entre a admissão hospitalar e o parto (48,6 vs. 16 dias; p=0,016). A taxa de mortalidade fetal foi menor no grupo cerclagem (5,3% vs. 54,5%, p=0,004). Considerando a idade gestacional no nascimento dos recém-nascidos vivos, não houve diferença entre os grupos cerclagem e expectante (29,13 vs. 27,4 semanas; p=0,857). CONCLUSÕES: A cerclagem de emergência foi associada a maior período de latência com impacto significativo na idade gestacional do parto e à redução da taxa de mortalidade fetal.

A case of sporotrichosis caused by different Sporothrix brasiliensis strains: mycological, molecular, and virulence analyses

BACKGROUND Sporotrichosis is a subcutaneous mycosis caused by dimorphic pathogenic fungi belonging to the Sporothrix genus. Pathogenic Sporothrix species typically produce melanin, which is known to be a virulence factor. OBJECTIVES The aim of this study was to perform phenotypic, genotypic, and virulence analyses of two distinct Sporothrix brasiliensis strains isolated from the same lesion on a patient from Rio de Janeiro. METHODS AND FINDINGS Genotypic analyses by partial sequencing of the calmodulin, β-tubulin, and chitin synthase genes, as well as polymerase chain reaction (PCR)-fingerprinting by T3B, M13, and GACA, showed that the isolates were very similar but not identical. Both isolates had similar phenotypic characteristics and effectively produced melanin in their yeast forms, accounting for their ability of causing disease in a murine sporotrichosis model. Remarkably, isolate B was albino in its environmental form but caused more severe disease than the pigmented A isolate. CONCLUSIONS These findings indicate that the patient was infected by two genetically and biologically distinct S. brasiliensis that vary in their production of melanin in their environmental forms. The results underscore the importance of characterizing phenotypically different isolates found in the same clinical specimen or patient.

Hepatite B e C em gestantes atendidas em um programa de pré-natal em um hospital universitário no Rio de Janeiro, Brasil: estudo retrospectivo de rastreamento da soroprevalência / Hepatitis b and c in pregnant women attended by a prenatal program in an universitary hospital in rio de janeiro, brazil: retrospective study of seroprevalence screening

ABSTRACT BACKGROUND: Hepatitis B and C are diseases with high morbimortality and constitute a global public health problem. In Brazil, the prevalence is not homogeneous, oscillating among different regions, but it is estimated that currently about 1% of the population present chronic disease related to the B virus and that there are 1.5 million infected with the C virus. Despite the development of hepatitis B vaccine, improvement in diagnostic methods and therapeutic advances in the field of viral hepatitis, there is still a large number of people who continues to be infected by these viruses, especially in populations at risk and also due to several factors, including vaccination and migration policies. Vertical and perinatal transmissions are of great importance in the epidemiology of viral hepatitis and the blood tests performed during prenatal care constitute a great opportunity for screening and identifying these viruses. OBJECTIVE: To evaluate the seroprevalence of markers for B and C viruses in women who underwent prenatal care at the Hospital Universitário Antônio Pedro (Antonio Pedro University Hospital) from 2006 to 2013 and to compare the results found with regional data and those described in the specific literature. METHODS: A descriptive, cross-sectional, quantitative study with retrospective data collected from 635 records of pregnant women attended at the Prenatal Service of the Hospital Universitário Antônio Pedro, Niterói, state of Rio de Janeiro, from March 2006 until December 2013. The database was built in the Microsoft Office Access program and was later exported to Microsoft Office Excel. For the processing and analysis of the data, it was used the SPSS (Statistical Package for Social Science, IBM) version 22.0, for Windows. RESULTS: Twelve cases with positive HBsAg (1.9%), 189 cases with positive anti-HBs (35.9%) and seven positive anti-HCV patients (1.3%) were observed. There was no significant association between age and positivity for HBsAg, anti-HBs and anti-HCV (P =0.205, 0.872 and 0.676, respectively). There was a direct relationship between the anti-HBs positivity and the last four years of the study (P<0.0001). CONCLUSION: A high prevalence of HBsAg was observed, higher than the expected for the evaluated region; there was a prevalence of anti-HCV, consistent with the current Brazilian reality; and a likely low rate of hepatitis B immunization, with a relatively high rate of susceptibility to this infection and no case of co-infection between B and C viruses and HIV. It is emphasized not only the need to trace hepatitis B and C, without exceptions, during prenatal care, since even though the current advances in therapy may not cure, at least they may allow a better quality of life for patients with chronic disease and the mandatory completion of immunoprophylaxis in all newborns. Special attention should be given to those patients susceptible to HBV, with prompt diagnosis and referral for specific vaccination.

RESUMO CONTEXTO: As hepatites pelo vírus B e C são doenças com elevada morbimortalidade e um problema de saúde pública global. No Brasil a prevalência não é homogênea, variando entre as diferentes regiões, mas estima-se que atualmente cerca de 1% da população apresente doença crônica relacionada ao vírus B e que haja 1,5 milhões de infectados pelo vírus C. Apesar do desenvolvimento da vacina contra a hepatite B, da melhoria nos métodos diagnósticos e dos avanços terapêuticos no campo das hepatites virais, ainda é grande o número de pessoas que continuam sendo infectadas por esses vírus, principalmente nas populações sob algum tipo de risco e devido a vários fatores incluindo políticas de vacinação e migração. A transmissão vertical e também a perinatal têm grande importância na epidemiologia das hepatites virais e os exames realizados durante o pré-natal constituem uma oportunidade única de rastreio e identificação destes vírus. OBJETIVO: Avaliar a soroprevalência de marcadores para os vírus B e C em mulheres que realizaram a assistência pré-natal no Hospital Universitário Antônio Pedro no período de 2006 a 2013 e comparar os resultados encontrados com os dados regionais e os descritos na literatura específica. MÉTODOS: Estudo transversal, descritivo, do tipo quantitativo, com coleta retrospectiva de dados em 635 prontuários de gestantes atendidas no Serviço de pré-natal do Hospital Universitário Antônio Pedro, Niterói, estado do Rio de Janeiro no período de março de 2006 a dezembro de 2013. O banco de dados foi construído no programa Microsoft Office Access, sendo posteriormente exportado para Microsoft Office Excel. Para o processamento e análise dos dados, foi utilizado o pacote estatístico SPSS (Statistical Package for Social Science, IBM) versão 22.0, para Windows. RESULTADOS: Foram observados 12 casos com HBsAg positivo (1,9%), 189 casos com anti-HBs positivo (35,9%) e sete pacientes positivas para o anti-HCV (1,3%). Não foi observada associação significativa entre a faixa etária e a positividade do HBsAg, anti-HBs e anti-HCV (P=0, 205, 0,872 e 0,676 respectivamente). Houve relação direta entre a positividade do anti-HBs e os últimos quatro anos da pesquisa (P<0,0001). CONCLUSÃO: Foi observada uma prevalência alta do HBsAg, acima daquela esperada para a região avaliada; uma prevalência para o anti-HCV concordante com a realidade brasileira atual; um índice provavelmente baixo de imunização contra a hepatite B, com índice relativamente alto de susceptibilidade para esta infecção e nenhum caso de coinfecção entre o vírus B, C e o HIV. Enfatiza-se não só a necessidade da triagem das hepatites B e C, sem exceções, durante o pré-natal, já que os avanços atuais na terapêutica poderão se não curar, pelo menos possibilitar uma melhor qualidade de vida para as pacientes com doença crônica e da realização mandatória da imunoprofilaxia em todos os recém-natos. Atenção especial deverá ser dada àquelas pacientes susceptíveis ao HBV, com pronto diagnóstico e encaminhamento para a realização da vacinação específica.

AKT as a key target for growth promoting functions of neutral ceramidase in colon cancer cells.

Despite advances in the field, colorectal cancer (CRC) remains a leading cause of cancer-related mortality worldwide. Research into bioactive sphingolipids over the past two decades has played an important role in increasing our understanding of the pathogenesis and therapeutics of CRC. In the complex metabolic network of sphingolipids, ceramidases (CDases) have a key function. These enzymes hydrolyze ceramides into sphingosine (SPH) which in turn is phosphorylated by sphingosine kinases (SK) 1 and 2 to generate sphingosine-1 phosphate (S1P). Importantly, we have recently shown that inhibition of neutral CDase (nCDase) induces an increase of ceramide in colon cancer cells which decreases cellular growth, increases apoptosis and modulates the WNT/ß-catenin pathway. We have also shown that the deletion of nCDase protected mice from the onset and progression of colorectal cancer in the AOM carcinogen model. Here, we demonstrate that AKT is a key target for the growth suppressing functions of ceramide. The results show that inhibition of nCDase activates GSK3ß through dephosphorylation, and thus is required for the subsequent phosphorylation and degradation of ß-catenin. Our findings show that inhibition of nCDase also inhibits the basal activation status of AKT, and we further establish that a constitutively active AKT (AKT T308D, S473D; AKTDD) reverses the effect of nCDase on ß-catenin degradation. Functionally, the AKTDD mutant is able to overcome the growth suppressive effects of nCDase inhibition in CRC cells. Moreover, nCDase inhibition induces a growth delay of xenograft tumors from control cells, whereas xenograft tumors from constitutively active AKT cells become resistant to nCDase inhibition. Taken together, these results provide important mechanistic insight into how nCDase regulates cell proliferation. These findings demonstrate a heretofore unappreciated, but critical, role for nCDase in enabling/maintaining basal activation of AKT and also suggest that nCDase is a suitable novel target for colon cancer therapy.

Alkaline ceramidase 2 is essential for the homeostasis of plasma sphingoid bases and their phosphates.

Sphingosine-1-phosphate (S1P) plays important roles in cardiovascular development and immunity. S1P is abundant in plasma because erythrocytes-the major source of S1P-lack any S1P-degrading activity; however, much remains unclear about the source of the plasma S1P precursor, sphingosine (SPH), derived mainly from the hydrolysis of ceramides by the action of ceramidases that are encoded by 5 distinct genes, acid ceramidase 1 ( ASAH1)/ Asah1, ASAH2/ Asah2, alkaline ceramidase 1 ( ACER1)/ Acer1, ACER2/ Acer2, and ACER3/ Acer3, in humans/mice. Previous studies have reported that knocking out Asah1 or Asah2 failed to reduce plasma SPH and S1P levels in mice. In this study, we show that knocking out Acer1 or Acer3 also failed to reduce the blood levels of SPH or S1P in mice. In contrast, knocking out Acer2 from either whole-body or the hematopoietic lineage markedly decreased the blood levels of SPH and S1P in mice. Of interest, knocking out Acer2 from whole-body or the hematopoietic lineage also markedly decreased the levels of dihydrosphingosine (dhSPH) and dihydrosphingosine-1-phosphate (dhS1P) in blood. Taken together, these results suggest that ACER2 plays a key role in the maintenance of high plasma levels of sphingoid base-1-phosphates-S1P and dhS1P-by controlling the generation of sphingoid bases-SPH and dhSPH-in hematopoietic cells.-Li, F., Xu, R., Low, B. E., Lin, C.-L., Garcia-Barros, M., Schrandt, J., Mileva, I., Snider, A., Luo, C. K., Jiang, X.-C., Li, M.-S., Hannun, Y. A., Obeid, L. M., Wiles, M. V., Mao, C. Alkaline ceramidase 2 is essential for the homeostasis of plasma sphingoid bases and their phosphates.

Tumor suppressor p53 links ceramide metabolism to DNA damage response through alkaline ceramidase 2.

p53 mediates the DNA damage response (DDR) by regulating the expression of genes implicated in cell cycle arrest, senescence, programmed cell death (PCD), and metabolism. Herein we demonstrate that human alkaline ceramidase 2 (ACER2) is a novel transcriptional target of p53 and that its transactivation by p53 mediates the DDR. We found that p53 overexpression or its activation by ionizing radiation (IR) upregulated ACER2 in cells. Two putative p53 responsive elements (p53REs) were found in its first intron of the ACER2 gene, and Chromatin Immunoprecipitation (ChIP) assays in combination with promoter activity assays demonstrated that these p53REs are the bona fide p53 binding sites that mediate ACER2 transactivation by p53. As ACER2 catalyzes the hydrolysis of ceramides into sphingosine, which in turn is phosphorylated to form sphingosine-1-phosphate (S1P), ACER2 upregulation increased the levels of both sphingosine and S1P while decreasing the levels of ceramides in cells. A moderate upregulation of ACER2 inhibited cell cycle arrest and cellular senescence in response to low-level expression of p53 or low-dose IR by elevating S1P, a pro-proliferative and pro-survival bioactive lipid, and/or decreasing ceramides whereas its robust upregulation mediated PCD in response to high-level expression of p53 or high-dose IR likely by accumulating cellular sphingosine, a pro-death bioactive lipid. ACER2 is frequently inactivated in various cancers due to its deletion or mutations, and restoring its expression inhibited the growth of tumor xenografts in mice. These results suggest that p53 mediates DDR and exerts its tumor suppressive role in part by regulating the expression of ACER2, which in turn regulates the bioactive sphingolipid lipids.

Targeting acid sphingomyelinase with anti-angiogenic chemotherapy.

Despite great promise, combining anti-angiogenic and conventional anti-cancer drugs has produced limited therapeutic benefit in clinical trials, presumably because mechanisms of anti-angiogenic tissue response remain only partially understood. Here we define a new paradigm, in which anti-angiogenic drugs can be used to chemosensitize tumors by targeting the endothelial acid sphingomyelinase (ASMase) signal transduction pathway. We demonstrate that paclitaxel and etoposide, but not cisplatin, confer ASMase-mediated endothelial injury within minutes. This rapid reaction is required for human HCT-116 colon cancer xenograft complete response and growth delay. Whereas VEGF inhibits ASMase, anti-VEGFR2 antibodies de-repress ASMase, enhancing endothelial apoptosis and drug-induced tumor response in asmase+/+, but not in asmase-/-, hosts. Such chemosensitization occurs only if the anti-angiogenic drug is delivered 1-2h before chemotherapy, but at no other time prior to or post chemotherapy. Our studies suggest that precisely-timed administration of anti-angiogenic drugs in combination with ASMase-targeting anti-cancer drugs is likely to optimize anti-tumor effects of systemic chemotherapy. This strategy warrants evaluation in future clinical trials.

Temporal and spatial evolution of maternal and neonatal mortality rates in Brazil, 1997-2012 / Evolução temporal e espacial das taxas de mortalidade materna e neonatal no Brasil, 1997-2012

Abstract Objective: Maternal and neonatal mortality are important public health issues in low-income countries. This study evaluated spatial and temporal maternal and neonatal mortality trends in Brazil between 1997 and 2012. Methods: This study employed spatial analysis techniques using death records from the mortality information system. Maternal mortality rates per 100,000 and neonatal mortality rates (early and late) per 1000 live births were calculated by state, region, and period (1997-2000, 2001-2004, 2005-2008, and 2009-2012). Multivariate negative binomial models were used to explain the risk of death. Results: The mean Brazilian maternal mortality rate was 55.63/100,000 for the entire 1997-2012 period. The rate fell 10% from 1997-2000 (58.92/100,000) to 2001-2004 (52.77/100,000), but later increased 11% during 2009-2012 (58.69/100,000). Early and late neonatal mortality rates fell 33% (to 7.36/1000) and 21% (to 2.29/1000), respectively, during the 1997-2012 period. Every Brazilian region witnessed a drop in neonatal mortality rates. However, maternal mortality increased in the Northeast, North, and Southeast regions. Conclusion: Brazil's neonatal mortality rate has improved in recent times, but maternal mortality rates have stagnated, failing to meet the Millennium Development Goals. Public policies and intersectoral efforts may contribute to improvements in these health indicators.

Resumo Objetivo: A mortalidade materna e neonatal é um importante problema de saúde pública em países de baixa renda. Este estudo avaliou as tendências de mortalidade materna e neonatal espacial e temporal no Brasil entre 1997 e 2012. Métodos: Este estudo usou técnicas de análise espacial com registros de óbito do Sistema de Informações sobre Mortalidade. As taxas de mortalidade materna a cada 100.000 e as taxas de mortalidade neonatal (precoce e tardia) a cada 1.000 nascidos vivos foram calculadas por estado, região e período (1997-2000, 2001-2004, 2005-2008 e 2009-2012). Os modelos binomiais negativos multivariados foram usados para explicar o risco de morte. Resultados: A taxa de mortalidade materna no Brasil foi 55,63/100.000 em todo o período entre 1997 e 2012. A taxa caiu 10% de 1997-2000 (58,92/100.000) a 2001-2004 (52,77/100.000), porém, mais tarde, aumentou 11% até 2009-2012 (58,69/100.000). As taxas de mortalidade neonatal precoce e tardia caíram 33% (para 7,36/1.000) e 21% (para 2.29/1.000), respectivamente, 1997-2012. Toda região brasileira testemunhou uma queda nas taxas de mortalidade neonatal. Contudo, a mortalidade materna aumentou nas regiões Nordeste, Norte e Sudeste. Conclusão: A taxa de mortalidade neonatal do Brasil melhorou nos últimos anos, porém as taxas de mortalidade materna estagnaram, deixaram de atingir os Objetivos de Desenvolvimento do Milênio. As políticas públicas e os esforços intersetoriais poderão contribuir para as melhorias nesses indicadores de saúde.

Role of neutral ceramidase in colon cancer.

Alterations in sphingolipid metabolism, especially ceramide and sphingosine 1-phosphate, have been linked to colon cancer, suggesting that enzymes of sphingolipid metabolism may emerge as novel regulators and targets in colon cancer. Neutral ceramidase (nCDase), a key enzyme in sphingolipid metabolism that hydrolyzes ceramide into sphingosine, is highly expressed in the intestine; however, its role in colon cancer has not been defined. Here we show that molecular and pharmacological inhibition of nCDase in colon cancer cells increases ceramide, and this is accompanied by decreased cell survival and increased apoptosis and autophagy, with minimal effects on noncancerous cells. Inhibition of nCDase resulted in loss of ß-catenin and inhibition of ERK, components of pathways relevant for colon cancer development. Furthermore, inhibition of nCDase in a xenograft model delayed tumor growth and increased ceramide while decreasing proliferation. It is noteworthy that mice lacking nCDase treated with azoxymethane were protected from tumor formation. Taken together, these studies show that nCDase is pivotal for regulating initiation and development of colon cancer, and these data suggest that this enzyme is a suitable and novel target for colon cancer therapy.-García-Barros, M., Coant, N., Kawamori, T., Wada, M., Snider, A. J., Truman, J.-P., Wu, B. X., Furuya, H., Clarke, C. J., Bialkowska, A. B., Ghaleb, A., Yang, V. W., Obeid, L. M., Hannun, Y. A. Role of neutral ceramidase in colon cancer.

ML264, A Novel Small-Molecule Compound That Potently Inhibits Growth of Colorectal Cancer.

Colorectal cancer is one of the leading causes of cancer mortality in Western civilization. Studies have shown that colorectal cancer arises as a consequence of the modification of genes that regulate important cellular functions. Deregulation of the WNT and RAS/MAPK/PI3K signaling pathways has been shown to be important in the early stages of colorectal cancer development and progression. Krüppel-like factor 5 (KLF5) is a transcription factor that is highly expressed in the proliferating intestinal crypt epithelial cells. Previously, we showed that KLF5 is a mediator of RAS/MAPK and WNT signaling pathways under homeostatic conditions and that it promotes their tumorigenic functions during the development and progression of intestinal adenomas. Recently, using an ultrahigh-throughput screening approach we identified a number of novel small molecules that have the potential to provide therapeutic benefits for colorectal cancer by targeting KLF5 expression. In the current study, we show that an improved analogue of one of these screening hits, ML264, potently inhibits proliferation of colorectal cancer cells in vitro through modifications of the cell-cycle profile. Moreover, in an established xenograft mouse model of colon cancer, we demonstrate that ML264 efficiently inhibits growth of the tumor within 5 days of treatment. We show that this effect is caused by a significant reduction in proliferation and that ML264 potently inhibits the expression of KLF5 and EGR1, a transcriptional activator of KLF5. These findings demonstrate that ML264, or an analogue, may hold a promise as a novel therapeutic agent to curb the development and progression of colorectal cancer.

Spatial dynamics of AIDS incidence in the elderly in Rio de Janeiro, Brazil, 1997-2011 / Dinâmica espacial da incidência da AIDS em idosos no Rio de Janeiro, Brasil, 1997-2011 / Dinámicas espaciales de la incidencia del SIDA en adultos de edad avanzada en Río de Janeiro, Brasil, 1997-2011

The dynamics of the spread of the AIDS epidemic ranges according to the characteristics of each geographical region in different population groups. The aim of this study was to evaluate spatial and temporal trends of the AIDS epidemic among the elderly in the State of Rio de Janeiro, Brazil. A retrospective study using spatial analysis techniques was conducted among AIDS cases (≥ 60 years) diagnosed from 1997-2011. The Poisson regression model was used to assess the relationship between year of diagnosis and incidence of AIDS, adjusted by sex. The AIDS epidemic began in the south coast of the state and gradually reached neighboring cities. The highest rates were found in regions around Rio de Janeiro and Niterói cities. The highest smoothed rates of the period were observed in Niterói in 2002-2006: 11.87/100,000 (men) and 8,5/100,000 (women). AIDS incidence rates among the elderly have stabilized in recent decades. To prevent HIV from spreading further among the general population, greater attention should be given to the older population.

A dinâmica de espalhamento da epidemia de AIDS varia segundo as características de cada região geográfica nos diferentes grupos populacionais. O objetivo deste trabalho foi avaliar tendências temporais e espaciais da epidemia de AIDS em idosos no Estado do Rio de Janeiro, Brasil. Estudo retrospectivo com técnicas de análise espacial, utilizando-se casos de AIDS (≥ 60 anos) diagnosticados de 1997-2011. O modelo de regressão de Poisson foi utilizado para acessar a relação entre ano diagnóstico e incidência de AIDS ajustada por sexo. A epidemia de AIDS começou no litoral sul do estado e, gradualmente, chegou às cidades vizinhas. As maiores taxas da doença foram encontradas em regiões em torno do Rio de Janeiro e Niterói. Em 2002-2006, na cidade de Niterói, foram observadas as maiores taxas suavizadas no período: 11,87/100 mil (homens) e 5,08/100 mil (mulheres). Os índices de AIDS em idosos têm estabilizado nas últimas décadas. Maior atenção deve ser dada ao grupo idoso para evitar a progressão da doença na população.

El objetivo de este estudio fue evaluar las tendencias espaciales y temporales de la epidemia del SIDA en los adultos de edad avanzada en el Estado de Río de Janeiro, Brasil. Se trata de un estudio retrospectivo de técnicas de análisis espacial, utilizando casos de SIDA (≥ 60 años) diagnosticados 1997-2011. El modelo de regresión de Poisson se utiliza para acceder a la relación entre año de diagnóstico y la incidencia del SIDA ajustada por sexo. La epidemia del SIDA comenzó en la costa sur del estado y, poco a poco, llegó a las ciudades aledañas. Se encontró que las tasas más altas de la enfermedad en las regiones que rodean Río de Janeiro y Niterói. En 2002-2006, en la ciudad de Niterói, hubo tasas más altas de la enfermedad suavizadas durante el período: 11,87/100.000 (hombres) y 8,5/100.000 (mujeres). Las tasas de SIDA en las personas mayores se han estabilizado en las últimas décadas. Se debe proporcionar una mayor atención al grupo de adultos de edad avanzada, con el fin de prevenir la progresión de la enfermedad en la población.

Profile of Brazilian smokers in the National Program for Tobacco Control.

OBJECTIVE: The treatment of tobacco addiction in Brazil has expanded in recent years; however, we must increase knowledge about the characteristics of individuals who adhere to cessation programs in order to adjust treatments to specific characteristics of the target population that favor success. The aim of this study was to describe the characteristics of smokers who present to Brazilian public health units seeking help to quit smoking based on the experience of a primary health care unit that covers a poor community in the city of Rio de Janeiro. METHODS: Data were collected at a Teaching Health Center from January 03 2012 to January 03 2014. RESULTS: Mean patient age was 49.32 ± 11.82 years, and 71% were women. About half of the participants successfully quit smoking (n=125, 51%). Higher levels of nicotine dependence were associated with lower levels of smoking cessation. There was a notable decrease in the probability of remaining smoking throughout the first month of treatment. After 3 weeks of treatment, only 19% had not quit smoking. The probability of quitting smoking decreases by 2% for every additional year of age. CONCLUSION: There is a need to revise and expand current strategies to make them more effective in preventing smoking since childhood.

Validation of a serodiagnostic test for sporotrichosis: a follow-up study of patients related to the Rio de Janeiro zoonotic outbreak.

The gold standard for the diagnosis of sporotrichosis consists of the isolation and identification of the fungus from clinical exudates and/or biopsy specimens. However, new technologies offer several advantages over the traditional methods because they are noninvasive and more sensitive in the differential diagnosis of infectious diseases. In the present study, we performed a validation, impact evaluation, and analysis of the applicability of an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of sporotrichosis in different groups of patients in comparison with the reference protocols for the evaluation of diagnostic tests for infectious diseases. We used 177 serum samples that were collected between 1998 and 2008 from patients in a geographic area related to the Rio de Janeiro outbreak of sporotrichosis. The ELISA had a low rate of cross-reactivity based on the reference values for diagnostic tests, and the analysis of the receiver operating characteristic curve revealed an area under the curve of 0.9154. Furthermore, higher accuracy was observed in the serodiagnosis of infections in special groups of patients such as pediatric and human immunodeficiency virus-positive patients. We concluded that this ELISA had a good clinical serological correlation and, therefore, can be considered a new diagnostic tool applicable to all clinical presentations of sporotrichosis.

As diferenças sexuais podem fundamentar estereótipos de gênero: deixem jovens de baixa escolaridade responderem / Can sex differences support gender stereotypes: let young people with low education answer

Os estereótipos de gênero configuram-se por crenças sobre o que caracteriza homens e mulheres. Tais crenças podem refletir nuances em diferenças sexuais que tiveram importância adaptativa para espécie e estão disseminadas na cultura. Este estudo buscou caracterizar crenças sobre homens e mulheres em população com baixa escolaridade a fim de mapear o conteúdo de estereótipos de gênero. A partir de respostas a um questionário aberto sobre características definidoras de homens e mulheres, delinearam-se 45 categorias de análise. Verificaram-sediscrepâncias significativas nas frequências de atribuições de categorias a homens e mulheres, bem como associações entre categorias e o sexo dos participantes. Aos homens foram atribuídas atividades de liderança e busca pelo sustento, eles foram caracterizados como pouco responsáveis e empáticos; as mulheres foram descritas como afetuosas e cuidadoras dos outros e de si mesmas. São discutidas relações entre os conteúdos das crenças estereotípicas e o conhecimento científico sobre diferenças sexuais.

Gender stereotypes are configured by beliefs about what characterizes men and women. These beliefs may reflect nuances on sexual differences that mattered to our species and are disseminated in the culture. This study sought to characterize beliefs about men and women in a sample with low schooling in order to map the content of gender stereotypes. From responses to an open-questionnaire about characteristics of men and women, we outlined 45 categories of analysis. We found significant discrepancies in frequencies of attribution of categories to men and women, as well as associations between categories and the sex of the participants. Men were associated with leadership activities and search for sustenance, less responsible and empathetic; women were described as affectionate and caring for others and of themselves. We discussed relations among the contents of stereotypic beliefs and scientific knowledge about sex differences.