Sit4 Genetically Interacts with Vps27 to Regulate Mitochondrial Function and Lifespan in Saccharomyces cerevisiae.

The Sit4 protein phosphatase plays a key role in orchestrating various cellular processes essential for maintaining cell viability during aging. We have previously shown that SIT4 deletion promotes vacuolar acidification, mitochondrial derepression, and oxidative stress resistance, increasing yeast chronological lifespan. In this study, we performed a proteomic analysis of isolated vacuoles and yeast genetic interaction analysis to unravel how Sit4 influences vacuolar and mitochondrial function. By employing high-resolution mass spectrometry, we show that sit4Δ vacuolar membranes were enriched in Vps27 and Hse1, two proteins that are part of the endosomal sorting complex required for transport-0. In addition, SIT4 exhibited a negative genetic interaction with VPS27, as sit4∆vps27∆ double mutants had a shortened lifespan compared to sit4∆ and vps27∆ single mutants. Our results also show that Vps27 did not increase sit4∆ lifespan by improving protein trafficking or vacuolar sorting pathways. However, Vps27 was critical for iron homeostasis and mitochondrial function in sit4∆ cells, as sit4∆vps27∆ double mutants exhibited high iron levels and impaired mitochondrial respiration. These findings show, for the first time, cross-talk between Sit4 and Vps27, providing new insights into the mechanisms governing chronological lifespan.

Beta-Adrenergic Blockade in Advanced Non-Small Cell Lung Cancer Patients Receiving Immunotherapy: A Multicentric Study.

Introduction The standard treatment of cancer has dramatically improved with immune checkpoint inhibitors (ICIs). Despite their proven advantage, many patients fail to exhibit a meaningful and lasting response. The beta-adrenergic signalling pathway may hold significant promise due to its role in promoting an immunosuppressive milieu within the tumour microenvironment. Inhibiting ß-adrenergic signalling could enhance ICI activity; however, blocking this pathway for this purpose has yielded conflicting results. The primary objective of this study was to evaluate the effect of beta-blocker use on overall survival and progression-free survival during ICI therapy. Methods A multicentric, retrospective, observational study was conducted in four Portuguese institutions. Patients with advanced non-small cell lung cancer treated with ICIs between January 2018 and December 2019 were included. Those using beta blockers for non-oncological reasons were compared with non-users. Results Among the 171 patients included, 36 concomitantly received beta blockers and ICIs. No significant increase was found in progression-free survival among patients who took ß-blockers (HR 0.74, 95% confidence interval (CI) 0.48-1.12, p = 0.151), and no statistically significant difference was found in overall survival. An apparent trend was observed towards better outcomes in the beta-blocker group, with a median overall survival of 9.93 months in the group not taking ß-blockers versus 14.90 months in the ß-blocker group (p = 0.291) and a median progression-free survival of 5.37 in the group not taking ß-blockers versus 10.87 months in the ß-blocker group (p = 0.151). Nine (25%) patients in the beta-blocker group and 16 (12%) in the non-beta-blocker group were progressive disease-free at the end of follow-up. This difference between the two groups is statistically significant (p = 0.047). Conclusion Our study found no statistically significant evidence that beta blockers enhance the effectiveness of immunotherapy. Using adrenergic blockade to modulate the immune system shows promise, warranting the need to develop prospective clinical studies.

Communication preferences and perceptions of cancer patient during their first medical oncology appointment.

BACKGROUND: The first medical oncology appointment serves as a platform for patients to comprehend their diagnosis and prognostic implications of cancer. This study aimed to determine patients' communication preferences during their first medical oncology appointment and to assess the disparities between patients' preferences and perceptions. METHODS: A total of 169 cancer patients participated by completing the Communication in First Medical Oncology Appointment Questionnaire (C-FAQ), a two-section questionnaire designed to assess patients' preferences and perceptions regarding Content (information provided and its extent), Facilitation (timing and location of information delivery), and Support (emotional support) during their first medical oncology appointment. A comparative analysis was conducted to assess the variations between preferences and perceptions. RESULTS: Content emerged as the most significant dimension compared to Facilitation and Support. The physician's knowledge, honesty, and ability to provide clear information were considered the most important attributes. Patients evaluated most of their preferences as "very important". Patients' perception of the communication dimensions present during their appointment was below preferences for 11 items, indicating significant discrepancies in clinical practice. CONCLUSIONS: Patients highly valued their preferences concerning Content, Facilitation, and Support dimensions of communication. However, patient preferences were more prominently oriented towards the Content dimension. The discrepancies between preferences and perceptions should be viewed as an opportunity for enhancing communication skills through training.

Iron Limitation Restores Autophagy and Increases Lifespan in the Yeast Model of Niemann-Pick Type C1.

Niemann-Pick type C1 (NPC1) is an endolysosomal transmembrane protein involved in the export of cholesterol and sphingolipids to other cellular compartments such as the endoplasmic reticulum and plasma membrane. NPC1 loss of function is the major cause of NPC disease, a rare lysosomal storage disorder characterized by an abnormal accumulation of lipids in the late endosomal/lysosomal network, mitochondrial dysfunction, and impaired autophagy. NPC phenotypes are conserved in yeast lacking Ncr1, an orthologue of human NPC1, leading to premature aging. Herein, we performed a phosphoproteomic analysis to investigate the effect of Ncr1 loss on cellular functions mediated by the yeast lysosome-like vacuoles. Our results revealed changes in vacuolar membrane proteins that are associated mostly with vesicle biology (fusion, transport, organization), autophagy, and ion homeostasis, including iron, manganese, and calcium. Consistently, the cytoplasm to vacuole targeting (Cvt) pathway was increased in ncr1∆ cells and autophagy was compromised despite TORC1 inhibition. Moreover, ncr1∆ cells exhibited iron overload mediated by the low-iron sensing transcription factor Aft1. Iron deprivation restored the autophagic flux of ncr1∆ cells and increased its chronological lifespan and oxidative stress resistance. These results implicate iron overload on autophagy impairment, oxidative stress sensitivity, and cell death in the yeast model of NPC1.

Mitochondrial respiration promotes Cdc37-dependent stability of the Cdk1 homolog Cdc28.

Cdc28, the homolog of mammalian Cdk1, is a conserved key regulatory kinase for all major cell cycle transitions in yeast. We have found that defects in mitochondrial respiration (including deletion of ATP2, an ATP synthase subunit) inhibit growth of cells carrying a degron allele of Cdc28 (cdc28td) or Cdc28 temperature-sensitive mutations (cdc28-1 and cdc28-1N) at semi-permissive temperatures. Loss of cell proliferation in the atp2Δcdc28td double mutant is associated with aggravated cell cycle arrest and mitochondrial dysfunction, including mitochondrial hyperpolarization and fragmentation. Unexpectedly, in mutants defective in mitochondrial respiration, steady-state protein levels of mutant cdc28 are strongly reduced, accounting for the aggravated growth defects. Stability of Cdc28 is promoted by the Hsp90-Cdc37 chaperone complex. Our results show that atp2Δcdc28td double-mutant cells, but not single mutants, are sensitive to chemical inhibition of the Hsp90-Cdc37 complex, and exhibit reduced levels of additional Hsp90-Cdc37 client kinases, suggesting an inhibition of this complex. In agreement, overexpression of CDC37 improved atp2Δcdc28td cell growth and Cdc28 levels. Overall, our study shows that simultaneous disturbance of mitochondrial respiration and Cdc28 activity reduces the capacity of Cdc37 to chaperone client kinases, leading to growth arrest.

Phosphoregulation of the ATP synthase beta subunit stimulates mitochondrial activity for G2/M progression.

Mitochondrial ATP synthase is a multifunctional enzyme complex involved in ATP production. We previously reported that the ATP synthase catalytic beta subunit (Atp2p in yeast) is regulated by the 2A-like protein phosphatase Sit4p, which targets Atp2p at T124/T317 impacting on ATP synthase levels and mitochondrial respiration. Here we report that Atp2-T124/T317 is also potentially regulated by Cdc5p, a polo-like mitotic kinase. Since both Cdc5p and Sit4p have established roles in cell cycle regulation, we investigated whether Atp2-T124/T317 phosphorylation was cell cycle-related. We present evidence that Atp2p levels and phosphorylation vary during cell cycle progression, with an increase at G2/M phase. Atp2-T124/T317 phosphorylation stimulates mitochondrial membrane potential, respiration and ATP levels at G2/M phase, indicating that dynamic Atp2p phosphorylation contributes to mitochondrial activity at this specific cell cycle phase. Preventing Atp2p phosphorylation delays G2/M to G1 transition, suggesting that enhanced bioenergetics at G2/M may help meet the energetic demands of cell cycle progression. However, mimicking constitutive T124/T317 phosphorylation or overexpressing Atp2p leads to mitochondrial DNA instability, indicating that reversible Atp2p phosphorylation is critical for homeostasis. These results indicate that transient phosphorylation of Atp2p, a protein at the core of the ATP production machinery, impacts on mitochondrial bioenergetics and supports cell cycle progression at G2/M.

Target of Rapamycin Complex 1 (TORC1), Protein Kinase A (PKA) and Cytosolic pH Regulate a Transcriptional Circuit for Lipid Droplet Formation.

Lipid droplets (LDs) are ubiquitous organelles that fulfill essential roles in response to metabolic cues. The identification of several neutral lipid synthesizing and regulatory protein complexes have propelled significant advance on the mechanisms of LD biogenesis in the endoplasmic reticulum (ER). However, our understanding of signaling networks, especially transcriptional mechanisms, regulating membrane biogenesis is very limited. Here, we show that the nutrient-sensing Target of Rapamycin Complex 1 (TORC1) regulates LD formation at a transcriptional level, by targeting DGA1 expression, in a Sit4-, Mks1-, and Sfp1-dependent manner. We show that cytosolic pH (pHc), co-regulated by the plasma membrane H+-ATPase Pma1 and the vacuolar ATPase (V-ATPase), acts as a second messenger, upstream of protein kinase A (PKA), to adjust the localization and activity of the major transcription factor repressor Opi1, which in turn controls the metabolic switch between phospholipid metabolism and lipid storage. Together, this work delineates hitherto unknown molecular mechanisms that couple nutrient availability and pHc to LD formation through a transcriptional circuit regulated by major signaling transduction pathways.

Production of IL-1ß, IL-6 and CXCL8 by endometrium of crossbred heifers stimulated with various pathogen-associated molecular patterns.

Uterine bacterial infections are common during the post-partum period of dairy herds and, apparently, incidences in crossbred cattle are less than in Holsteins. The aims of this study were (I) to evaluate production of interleukin 1-ß (IL-1ß), interleukin-6 (IL-6) and chemokine CXCL8 using endometrial explants from Bos indicus crossbred heifers at diestrous, stimulated by various pathogen-associated molecular patterns (PAMP), and (II) assess production of these cytokines by lipopolysaccharide-stimulated endometrial explants from heifers when samples were collected at different stages of estrous cycle. In the first experiment, endometrial explants from heifers at diestrous were stimulated by ten-fold serial dilutions of lipopolysaccharide (LPS), triacylated lipopeptide (PAM3) or peptidoglycan (PGN). In the second experiment, endometrial explants collected at different stages of the estrous cycle were treated with LPS. Concentrations of IL-1ß, IL-6 and CXCL8 were quantified in supernatant. There was a marked (P < 0.05) production of IL-1ß, IL-6, and CXCL8 in response to LPS treatment. There was also production of IL-1ß (P < 0.05) in response to PGN treatment. Explant samples collected at different stages of the estrous cycle responded to LPS treatment with production of IL-1ß and IL-6, but with no differences (P > 0.05) between stages of estrous cycle. In conclusion, endometrial samples of crossbred Zebu-based heifers collected during diestrous produced IL-1ß, IL-6 and CXCL8 in response to LPS and IL-1ß in response to PGN. The cytokine production in response to LPS, however, was not affected by the stage of the estrous cycle in Bos indicus crossbred heifers.

Incentives and barriers to HIV testing among female sex workers in Ceará.

OBJECTIVE: Estimating HIV prevalence and describing the incentives and barriers for HIV testing among female sex workers. METHODS: This cross-sectional study recruited 402 women aged 18 years or older, residing in Fortaleza, state of Ceará, Brazil, who reported having had sexual intercourse in exchange for money in last four months. The sample was recruited using Respondent Driven Sampling, between August and November 2010. RESULTS: The 84.1% of the sample tested and the estimated prevalence of HIV infection was 3.8%. The sample was young (25 to 39 years ), single (80.0%), with one to three children (83.6%), had eight or more years of schooling (65.7%), and belonged to social classes D/E (53.1%). The majority worked in fixed locations (bars, motels, hotels, sauna - 88.9%), and prostitution was their only source of income (54.1%). About 25% of the sample did not know where to test in the public health sector and 51.8% either never tested or hadn't tested for over a year or more. The main reported barriers to testing were the perceptions that there was no risk of becoming infected (24.1%), and, alternatively, fear of discrimination if the test was positive (20.5%). Incentives for testing were the greater availability of testing sites (57.0%) and health facilities with alternative schedules (44.2%). CONCLUSIONS: Prevalence for HIV was similar to that found in other Brazilian cities in different regions of the country, although higher than the general female population. Non-traditional venues not associated with the health system and availability of testing in health units during non-commercial hours are factors that encourage testing. Not considering oneself to be at risk, fear of being discriminated against and not knowing testing locations are barriers.

Signaling pathways governing iron homeostasis in budding yeast.

Iron is an essential element for eukaryotes as it participates as a redox-active co-factor in many biological processes. Since iron is also potentially toxic, iron levels are carefully regulated. In the yeast Saccharomyces cerevisiae, iron homeostasis is maintained by the transcriptional control of the iron acquisition systems (iron regulon), mainly by the iron-responsive transcriptional factors Aft1p and Yap5p. Intracellular iron is stored in the vacuole, mobilized for other locations when necessary, particularly for the mitochondria, the major site of iron-utilizing pathways. Mitochondria also play an additional role as a sensor for the regulation of cellular iron acquisition and intracellular distribution. Mounting evidence suggest that iron acquisition systems are not only responsive to iron levels but also to signaling pathways. The most recognized is the activation of the iron regulon at the diauxic shift, oppositely regulated by PKA and SNF1 kinases, major regulators of glucose signaling. Hog1p, a MAP kinase involved in stress responses, also negatively regulates iron uptake by phosphorylating Aft1p. In this review, we address organellar signaling and signal transduction pathways that play a major role in the coordination of iron homeostasis with cell growth and division.

The ceramide activated protein phosphatase Sit4 impairs sphingolipid dynamics, mitochondrial function and lifespan in a yeast model of Niemann-Pick type C1.

The Niemann-Pick type C is a rare neurodegenerative disease that results from loss-of-function point mutations in NPC1 or NPC2, which affect the homeostasis of sphingolipids and sterols in human cells. We have previously shown that yeast lacking Ncr1, the orthologue of human NPC1 protein, display a premature ageing phenotype and higher sensitivity to oxidative stress associated with mitochondrial dysfunctions and accumulation of long chain bases. In this study, a lipidomic analysis revealed specific changes in the levels of ceramide species in ncr1Δ cells, including decreases in dihydroceramides and increases in phytoceramides. Moreover, the activation of Sit4, a ceramide-activated protein phosphatase, increased in ncr1Δ cells. Deletion of SIT4 or CDC55, its regulatory subunit, increased the chronological lifespan and hydrogen peroxide resistance of ncr1Δ cells and suppressed its mitochondrial defects. Notably, Sch9 and Pkh1-mediated phosphorylation of Sch9 decreased significantly in ncr1Δsit4Δ cells. These results suggest that phytoceramide accumulation and Sit4-dependent signaling mediate the mitochondrial dysfunction and shortened lifespan in the yeast model of Niemann-Pick type C1, in part through modulation of the Pkh1-Sch9 pathway.

The Hog1p kinase regulates Aft1p transcription factor to control iron accumulation.

Iron acquisition systems have to be tightly regulated to assure a continuous supply of iron, since it is essential for survival, but simultaneously to prevent iron overload that is toxic to the cells. In budding yeast, the low­iron sensing transcription factor Aft1p is a master regulator of the iron regulon. Our previous work revealed that bioactive sphingolipids modulate iron homeostasis as yeast cells lacking the sphingomyelinase Isc1p exhibit an upregulation of the iron regulon. In this study, we show that Isc1p impacts on iron accumulation and localization. Notably, Aft1p is activated in isc1Δ cells due to a decrease in its phosphorylation and an increase in its nuclear levels. Consistently, the expression of a phosphomimetic version of Aft1p-S210/S224 that favours its nuclear export abolished iron accumulation in isc1Δ cells. Notably, the Hog1p kinase, homologue of mammalian p38, interacts with and directly phosphorylates Aft1p at residues S210 and S224. However, Hog1p-Aft1p interaction decreases in isc1Δ cells, which likely contributes to Aft1p dephosphorylation and consequently to Aft1p activation and iron overload in isc1Δ cells. These results suggest that alterations in sphingolipid composition in isc1Δ cells may impact on iron homeostasis by disturbing the regulation of Aft1p by Hog1p. To our knowledge, Hog1p is the first kinase reported to directly regulate Aft1p, impacting on iron homeostasis.

Incentives and barriers to HIV testing among female sex workers in Ceará / Incentivos e barreiras ao teste de HIV entre mulheres profissionais do sexo no Ceará

ABSTRACT OBJECTIVE Estimating HIV prevalence and describing the incentives and barriers for HIV testing among female sex workers. METHODS This cross-sectional study recruited 402 women aged 18 years or older, residing in Fortaleza, state of Ceará, Brazil, who reported having had sexual intercourse in exchange for money in last four months. The sample was recruited using Respondent Driven Sampling, between August and November 2010. RESULTS The 84.1% of the sample tested and the estimated prevalence of HIV infection was 3.8%. The sample was young (25 to 39 years ), single (80.0%), with one to three children (83.6%), had eight or more years of schooling (65.7%), and belonged to social classes D/E (53.1%). The majority worked in fixed locations (bars, motels, hotels, sauna - 88.9%), and prostitution was their only source of income (54.1%). About 25% of the sample did not know where to test in the public health sector and 51.8% either never tested or hadn't tested for over a year or more. The main reported barriers to testing were the perceptions that there was no risk of becoming infected (24.1%), and, alternatively, fear of discrimination if the test was positive (20.5%). Incentives for testing were the greater availability of testing sites (57.0%) and health facilities with alternative schedules (44.2%). CONCLUSIONS Prevalence for HIV was similar to that found in other Brazilian cities in different regions of the country, although higher than the general female population. Non-traditional venues not associated with the health system and availability of testing in health units during non-commercial hours are factors that encourage testing. Not considering oneself to be at risk, fear of being discriminated against and not knowing testing locations are barriers.

RESUMO OBJETIVO Estimar a prevalência do HIV e descrever os incentivos e barreiras à realização do teste para o HIV entre mulheres profissionais do sexo. MÉTODOS Este estudo transversal recrutou 402 mulheres de 18 anos ou mais, residentes em Fortaleza, CE, que informaram ter tido relação sexual em troca de dinheiro nos últimos quatro meses. A amostra foi recrutada por meio da técnica Respondent Driven Sampling, entre agosto e novembro de 2010. RESULTADOS A adesão ao teste de HIV foi de 84,1% e a prevalência estimada da infecção pelo HIV foi de 3,8%. A amostra era jovem (25 a 39 anos), solteira (80,0%), com um a três filhos (83,6 %), tinham oito anos ou mais de estudo (65,7%) e pertencia às classes sociais D/E (53,1%). A maioria exercia a profissão em locais fechados (bares, motéis, hotéis, sauna - 88,9%), e a prostituição era a única fonte de renda (54,1%). Cerca de 25% da amostra desconhecia onde o teste de HIV era realizado na rede pública e 51,8% nunca fez o teste ou se testou há um ano ou mais. As principais barreiras ao teste foram acreditar que não corre risco de se infectar (24,1%) e o medo da discriminação caso o teste fosse reagente (20,5%). Os incentivos foram relacionados à maior oferta de locais para o teste (57,0%) e de unidades de saúde com horários alternativos (44,2%). CONCLUSÕES A prevalência foi semelhante à encontrada em outras cidades brasileiras de diferentes regiões do país, apesar de superiores a de mulheres não profissionais do sexo. A disponibilidade do teste em locais não relacionados à saúde e a oferta nas unidades básicas em horário não comercial são fatores que incentivam a realização do teste. Não se considerar sob-risco, medo de ser discriminada e desconhecimento dos locais onde o teste é realizado podem ser uma barreira para a realização do exame.

[Evaluation of implementation of the protocol for managing tuberculosis/ human immunodeficiency virus coinfection in specialized care services in ceará state]. / Evaluación de la implementación del protocolo de manejo de coinfección de tuberculosis y virus de inmunodeficiencia humana en los servicios de asistencia especializada del estado de Ceará.

OBJECTIVES: Identify barriers and strategies for implementation of the protocol for managing tuberculosis/ human immunodeficiency virus (TB/HIV) coinfection in specialized care services in Ceará state, through evaluation research. METHODS: The study followed iPIER methodology, a new initiative to help improve the implementation of health programs through the use of research that is conducted as a part of the implementation process. Data were collected on the structure and processes of 22 services and barriers were explored in four focus groups with 28 participants, from the standpoint of the health team, administrators, and users. The discussions were transcribed and interpreted with regard to the objectives of the study. RESULTS: The information on structure and process revealed that six services manage TB/HIV coinfection and 16 do not. The barriers were: team members were unaware of the protocol; lack of clinical practice guidelines in the services; specialized care services working at all three levels of the health system; inadequate spaces for treatment of airborne diseases; and lack of communication with primary health care sectors for patient transfers. The results were discussed with teams and administrators in seminars held in the services and with program managers for sexually transmitted infections, human immunodeficiency virus, AIDS, hepatitis, and tuberculosis. CONCLUSIONS: Direct dialogue between administrators, implementers, users, and researchers generated knowledge about the services and led to joint preparation of modifications in workflow aimed at acceptance and use of the protocol; however, users continue to resist adherence to treatment.

Hazardous alcohol use among transwomen in a Brazilian city.

There is a lack of information about alcohol use by transgender women. We estimated the prevalence of dangerous alcohol use in the last 12 months by transgender women, who are known as travestis in Brazil, and we identified the associated risk factors. Three hundred travestis were recruited using Respondent Driving Sampling (RDS). We applied the Alcohol Use Disorders Identification Test (AUDIT). We controlled the sample by applying a weight to each interviewee. Three quarters (74.2%) of travestis were regular drinkers, half (48.7%) scored over eight in the AUDIT and 14.8% scored over 20. The risk factors for alcohol use were: aged over 24, low income and unprotected sex. The dangerous use of alcohol is prevalent among travestis. Given that this group has a greater risk of HIV infection and transmission, and that the dangerous use of alcohol was associated with unsafe sex, specific intervention strategies are required.

Evaluación de la implementación del protocolo de manejo de coinfección de tuberculosis y virus de inmunodeficiencia humana en los servicios de asistencia especializada del estado de Ceará / Evaluation of implementation of the protocol for managing tuberculosis/ human immunodeficiency virus coinfection in specialized care services in ceará state

RESUMEN Objetivos Identificar barreras y estrategias para la implementación del protocolo “Manejo de coinfección por tuberculosis y virus de inmunodeficiencia humana (TBC/VIH)” en los servicios de asistencia especializada (SAE) de Ceará, mediante investigación evaluativa. Métodos Estudio desarrollado siguiendo la metodología iPIER, una nueva iniciativa con el propósito de apoyar mejoras en la ejecución de programas de salud a través de investigaciones integradas en ellos acerca de su ejecución. Se recogieron datos de estructura y procesos de 22 servicios y se exploraron las barreras mediante cuatro grupos focales, con 28 participantes, desde el punto de vista del equipo de salud, los administradores y los usuarios. Las discusiones fueron transcritas e interpretadas según los objetivos del estudio. Resultados Los datos de estructura y procesos revelaron que seis servicios realizan acciones de manejo de coinfección TB/VIH y 16, no lo hacen. Las barreras fueron: desconocimiento del protocolo en los equipos, ausencia de guías de práctica clínica en los servicios, inserción de los SAE en los tres niveles del sistema de salud, espacios inadecuados para tratar enfermedades de transmisión aérea y falta de comunicación con los sectores de atención primaria de salud para los traslados. Se discutieron los resultados con equipos y administradores en seminarios en los servicios y con los responsables de los programas de enfermedades de transmisión sexual, virus de inmunodeficiencia humana, sida, hepatitis y tuberculosis. Conclusiones El diálogo directo entre administradores, ejecutores, usuarios e investigadores generó conocimiento sobre los servicios y elaboración conjunta de modificaciones de flujos para la aceptación y utilización del protocolo; sin embargo, entre los usuarios persiste la resistencia para adherirse al tratamiento.

ABSTRACT Objectives Identify barriers and strategies for implementation of the protocol for managing tuberculosis/ human immunodeficiency virus (TB/HIV) coinfection in specialized care services in Ceará state, through evaluation research. Methods The study followed iPIER methodology, a new initiative to help improve the implementation of health programs through the use of research that is conducted as a part of the implementation process. Data were collected on the structure and processes of 22 services and barriers were explored in four focus groups with 28 participants, from the standpoint of the health team, administrators, and users. The discussions were transcribed and interpreted with regard to the objectives of the study. Results The information on structure and process revealed that six services manage TB/HIV coinfection and 16 do not. The barriers were: team members were unaware of the protocol; lack of clinical practice guidelines in the services; specialized care services working at all three levels of the health system; inadequate spaces for treatment of airborne diseases; and lack of communication with primary health care sectors for patient transfers. The results were discussed with teams and administrators in seminars held in the services and with program managers for sexually transmitted infections, human immunodeficiency virus, AIDS, hepatitis, and tuberculosis. Conclusions Direct dialogue between administrators, implementers, users, and researchers generated knowledge about the services and led to joint preparation of modifications in workflow aimed at acceptance and use of the protocol; however, users continue to resist adherence to treatment.

Hazardous alcohol use among transwomen in a Brazilian city / Consumo excessivo de álcool entre mulheres transgênero numa cidade brasileira / Consumo excesivo de alcohol entre mujeres transexuales en una ciudad brasileña

Abstract: There is a lack of information about alcohol use by transgender women. We estimated the prevalence of dangerous alcohol use in the last 12 months by transgender women, who are known as travestis in Brazil, and we identified the associated risk factors. Three hundred travestis were recruited using Respondent Driving Sampling (RDS). We applied the Alcohol Use Disorders Identification Test (AUDIT). We controlled the sample by applying a weight to each interviewee. Three quarters (74.2%) of travestis were regular drinkers, half (48.7%) scored over eight in the AUDIT and 14.8% scored over 20. The risk factors for alcohol use were: aged over 24, low income and unprotected sex. The dangerous use of alcohol is prevalent among travestis. Given that this group has a greater risk of HIV infection and transmission, and that the dangerous use of alcohol was associated with unsafe sex, specific intervention strategies are required.

Resumo: Informação sobre uso de álcool entre mulheres transgêneros são escassas. Estimamos a prevalência do uso perigoso de álcool nos últimos 12 meses entre mulheres transgêneros chamadas travestis no Brasil, e identificados os fatores de risco associados. Trezentas travestis foram recrutadas utilizando Respondent Driving Sampling (RDS). Aplicamos o Alcohol Use Disorders Identification Test (AUDIT). Controlamos a amostragem, com um peso aplicado a cada entrevistada. Três quartos (74,2%) das travestis eram bebedores regulares, metade (48,7%) obteve > 8 no AUDIT e 14,8% obtiveram > 20. Fatores de risco para o uso de risco de álcool foram: > 24 anos, baixa renda, raça negra, viver com a família, ter feito sexo por dinheiro, uso de drogas ilícitas nos últimos seis meses e sexo desprotegido. O uso perigoso de álcool é prevalente entre travestis. Tendo em vista que este grupo possui maior risco para a infecção e transmissão de HIV, e que o uso perigoso de álcool foi associado ao sexo inseguro, são necessárias estratégias de intervenção específicas.

Resumen: Existe falta de información sobre el consumo de alcohol por parte de mujeres transexuales. El estudio estimó la prevalencia de consumo excesivo de alcohol durante los últimos 12 meses por parte de mujeres transexuales, conocidas en Brasil como travestis, e identificó los factores de riesgo. Fueron reclutadas 300 travestis, utilizando la técnica de Respondent Driven Sampling (RDS). Aplicamos el Alcohol Use Disorders Identification Test (AUDIT). La muestra fue controlada, aplicando un peso a cada entrevistada. Tres cuartos (74,2%) de la muestra consumían alcohol regularmente, la mitad (48,7%) totalizó más de ocho puntos en el AUDIT y 14,8% sumaron más de 20 puntos. Los factores de riesgo para el consumo excesivo de alcohol fueron: edad por encima de 24 años, baja renta y sexo sin preservativo. El uso excesivo de alcohol es común entre las travestis. Debido a que el grupo presenta riesgo aumentado de transmisión del VIH, y que el uso excesivo de alcohol estuvo asociado al sexo inseguro, se necesitan estrategias específicas para mitigar los riesgos.

Changes in endometrial transcription of TLR2, TLR4, and CD14 during the first-week postpartum in dairy cows with retained placenta.

Changes in the endometrial transcription of pattern recognition receptors may increase the susceptibility to postpartum uterine infections in Holstein cows with retained placenta. To test this hypothesis, nine cows with retained placenta and ten cows without retained placenta were submitted to endometrial biopsies at the first and seventh days postpartum. Cows were monitored weekly with clinical and gynecological examinations until 42 days postpartum. Samples of the uterine contents were collected weekly for aerobic bacteria isolation. All cows had endometrial transcription of Toll-like receptors (TLRs) 1/6, 2, 4, 5, and 9; nucleotide-binding oligomerization domain (NOD)-like receptors 1 and 2; and the coreceptors cluster of differentiation 14 (CD14) and myeloid differentiation protein-2 (MD-2), as measured on the first and seventh days postpartum. Escherichia coli was the most common bacterium isolated from the uterine contents of cows with or without retained placenta until 21 days postpartum. Transcription levels of TLR2, TLR4, and CD14 in Holstein cows with retained placenta significantly decreased (P < 0.05) between the first and the seventh day postpartum. Conversely, cows without retained placenta did not have any significant changes in transcription levels between these time points.

Comparative proteomic analyses of urine from rat urothelial carcinoma chemically induced by exposure to N-butyl-N-(4-hydroxybutyl)-nitrosamine.

Bladder cancer is estimated to be the ninth most common malignancy with a high rate of recurrence and progression despite therapy, early diagnosis being crucial for timely intervention. Using a well-established animal model of urothelial carcinoma, we performed a comprehensive analysis of urine proteome profile from healthy animals and animals with urothelial carcinoma at two time-points of disease pathogenesis. GeLC-MS/MS, followed by bioinformatics analysis of unique proteins and the ones present in significantly distinct levels among groups, highlighted the biological processes involved in disease pathogenesis such as, for instance, response to selenium and to drugs, neutral lipid metabolism at earlier stages of disease, and inflammation, immune response and wound healing at advanced stages. Proteins from up-regulated biological processes might be seen as putative disease biomarkers. These include, for example, cadherins, lipoproteins, and glysosyltransferases, which may be included in multimarker strategies. Taken together, the data support the application of urine proteomics for the identification of the biological processes modulated by bladder cancer in an integrative perspective. The present exploratory urinary proteomic analysis might be seen as an important starting point for studies targeting urinary proteins in human, aiming at the implementation of novel laboratory approaches for the detection and successful management of urothelial carcinoma.

Molecular detection of haemotropic Mycoplasma species in urban and rural cats from Portugal.

OBJECTIVES: The aim of the present study was to evaluate the prevalence of haemoplasma infection in cats in Portugal and to assess risk factors for infection. METHODS: Real-time polymerase chain reaction techniques were used to assess 236 urban and rural cats from central and southern Portugal. RESULTS: The overall prevalence of haemoplasma in the target population was 27.1% (64/236), with individual species' prevalences as follows: 17.8% (42/236) 'Candidatus Mycoplasma haemominutum' (CMhm), 14.4% (34/236) Mycoplasma haemofelis (Mhf) and only 5.9% (14/236) 'Candidatus Mycoplasma turicensis' (CMt). Multiple infections were detected in 8.1% (19/236) of the samples, with triple and double infections with Mhf and CMhm being most commonly detected (5.9% [14/236] of cats). Haemoplasma infection was significantly higher in shelter cats (P = 0.015) than in cats with other lifestyles (eg, free-roaming/house pet/blood donors). Haemoplasma prevalence was also higher in cats with feline immunodeficiency virus infection (FIV; P = 0.011). Although sex was not significantly associated with haemoplasma infection (P = 0.050), CMt was predominantly found in males (P = 0.032). Also, the presence of haemoplasma multiple infections was statistically associated with being in a shelter (P = 0.021), male (P = 0.057) and with FIV co-infection (P = 0.004). No evidence of an association between haemoplasma infection and geographical location, age or feline leukaemia virus co-infection was found. CONCLUSIONS AND RELEVANCE: The results obtained in our study are consistent with the documented worldwide prevalence of feline haemoplasma infections, suggesting that the three main feline haemoplasma species are common in Portugal.