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Background: Childhood obesity has been growing steadily, at an earlier age, and currently comprises a public health issue. A number of studies have pointed to perinatal factors as possible determinants in the development of childhood obesity. Objective: To evaluate the influence of perinatal factors on the development of obesity in children and adolescents in southern Brazil. Design: Retrospective cohort study in which a linkage was made between anthropometric data of children and adolescents aged 0 to 15 years who had been registered in the Food and Nutrition Surveillance System (SISVAN) from 2008 to 2016 and their perinatal data registered in the Live Birth Information System (SINASC) from 2000 to 2014. The SINASC was used to extract maternal covariables (age, schooling, marital status), prenatal variables (parity and number of prenatal visits), and perinatal variables (type of delivery, sex, and birth weight). Variables such as age, inclusion in the Bolsa Família income transfer program, and the number of anthropometric evaluations were extracted from SISVAN. Results: The sample comprised 537 children and adolescents. The median age was 8 years (interquartile range: 2-11 years). The prevalence of obesity was 15.1%. Poisson regression revealed a higher risk of obesity in children born via cesarean delivery (relative risk [RR] = 1.48; 95% CI: 1.01-2.17), children of primiparous mothers (RR = 1.72; 95% CI: 1.16-2.53), girls (RR = 1.77; 95% CI:1.21-2.60), and those aged between 5 and 9 years (RR = 26.8; 95% CI: 3.75-191.55) and older than 10 years (RR = 20.74; 95% CI: 2.89-148.61). Conclusions: The linkage between SINASC and SISVAN allowed identification of prenatal and perinatal risk factors for the development of childhood obesity. These findings should contribute to the development of health promotion and prevention policies.

Background: Drug resistance is the main cause of therapy failure in advanced lung cancer. Although non-genetic mechanisms play important roles in tumor chemoresistance, drug-induced epigenetic reprogramming is still poorly understood. Materials and Methods: The A549 cell line was used to generate cells with non-genetic resistance to cisplatin (CDDP), namely A549/CDDP cells. Bioorthogonal non-canonical amino acid tagging (BONCAT) and mass spectrometry were used to identify proteins modulated by CDDP in A549 and A549/CDDP cells. Results: Proteins related to proteostasis, telomere maintenance, cell adhesion, cytoskeletal remodeling, and cell redox homeostasis were found enriched in both cell lines upon CDDP exposure. On the other hand, proteins involved in drug response, metabolic pathways and mRNA processing and splicing were up-regulated by CDDP only in A549/CDDP cells. Conclusion: Our study revealed proteome dynamics involved in the non-genetic response to CDDP, pointing out potential targets to monitor and overcome epigenetic resistance in lung cancer.

Dental wear is a multifactorial progressive condition of hard tissue loss, often decreasing the vertical dimension and leading to functional and esthetic consequences. Regaining space by using contemporary restorative materials has enabled less invasive treatment options. This clinical report describes a complete mouth restoration of a patient with severe generalized dental wear by progressively increasing the vertical dimension and using conservative prosthetic treatment options to maintain pulp vitality. This multidisciplinary approach is considered safe and predictable, providing long-term comfort and esthetics.

Erythema Nodosum Leprosum (ENL) is a recurrent acute inflammatory complication of leprosy affecting up to 50% of all Borderline Lepromatous and Lepromatous Leprosy (BL/LL) patients. Although ENL is described as an immune reaction mediated by neutrophils, studies demonstrating the direct role of neutrophils in ENL are still rare. One subpopulation of low-density neutrophils (LDNs), present within the fraction of peripheral blood mononuclear cells (PBMC), has been associated with the pathogenesis and severity of diseases like sepsis, lupus, and tuberculosis. We herein analyzed LDNs and high-density neutrophils (HDNs) in terms of frequency, phenotype, and morphology. Serum levels of MMP-9 (a neutrophilic degranulation marker) were evaluated by ELISA; and LDNs were generated in vitro by stimulating healthy-donor, whole-blood cultures. PBMC layers of ENL patients presented segmented/hypersegmented cells that were morphologically compatible with neutrophils. Immunofluorescence analyses identified LDNs in ENL. Flow cytometry confirmed the elevated frequency of circulating LDNs (CD14−CD15+) in ENL patients compared to healthy donors and nonreactional Borderline Tuberculoid (BT) patients. Moreover, flow cytometry analyses revealed that ENL LDNs had a neutrophilic-activated phenotype. ENL patients under thalidomide treatment presented similar frequency of LDNs as observed before treatment but its activation status was lower. In addition, Mycobacterium leprae induced in vitro generation of LDNs in whole blood in a dose-dependent fashion; and TGF-β, an inhibitor of neutrophilic degranulation, prevented LDNs generation. MMP-9 serum levels of BL/LL patients with or without ENL correlated with LDNs frequency at the same time that ultrastructural observations of ENL LDNs showed suggestive signs of degranulation. Together, our data provide new insights into the knowledge and understanding of the pathogenesis of ENL while enriching the role of neutrophils in leprosy.

The aim of this study was to establish a peptidomic profile based on LC-MS/MS and random forest (RF) algorithm to distinguish the urinary peptidomic scenario of type 2 diabetes mellitus (T2DM) patients with different stages of diabetic kidney disease (DKD). Urine from 60 T2DM patients was collected: 22 normal (stage A1), 18 moderately increased (stage A2) and 20 severely increased (stage A3) albuminuria. A total of 1080 naturally occurring peptides were detected, which resulted in the identification of a total of 100 proteins, irrespective of the patients’ renal status. The classification accuracy showed that the most severe DKD (A3) presented a distinct urinary peptidomic pattern. Estimates for peptide importance assessed during RF model training included multiple fragments of collagen and alpha-1 antitrypsin, previously associated to DKD. Proteasix tool predicted 48 proteases potentially involved in the generation of the 60 most important peptides identified in the urine of DM patients, including metallopeptidases, cathepsins, and calpains. Collectively, our study lightened some biomarkers possibly involved in the pathogenic mechanisms of DKD, suggesting that peptidomics is a valuable tool for identifying the molecular mechanisms underpinning the disease and thus novel therapeutic targets.

Resumo O objetivo deste artigo é verificar a associação entre fatores maternos e antropométricos e o consumo de alimentos ultraprocessados em crianças de 4 a 24 meses de idade. Métodos: Estudo transversal, com 300 crianças internadas em um hospital terciário e suas mães. A entrevista deu-se nas primeiras 72 horas de internação para evitar interferência nas respostas sobre a alimentação da criança. Os fatores maternos investigados foram: idade, escolaridade, renda, paridade, IMC e orientação sobre alimentação complementar. As variáveis referentes às crianças investigadas foram: idade, aleitamento materno, escola infantil, IMC/idade, estatura/idade, peso/idade e introdução de alimentos ultraprocessados. A associação entre os fatores estudados e a introdução de alimentos ultraprocessados foi testada por regressão linear. O nível de significância considerado foi de 0.05. Verificou-se que apenas 21% das crianças ainda não haviam recebido nenhum tipo de alimento ultraprocessado, sendo que 56.5% recebeu algum destes alimentos antes dos seis meses. Na análise multivariada, escolaridade materna, renda familiar, idade materna e paridade foram associadas à oferta de alimentos ultraprocessados. As práticas alimentares de crianças entre 4 e 24 meses estão inadequadas frente às recomendações para a faixa etária. Abstract Objective To verify the association of maternal and anthropometric factors with consumption of ultra-processed foods in children between 4 to 24 months. Methods cross-sectional study with 300 children hospitalized in a tertiary hospital and their mothers. The interview took place during the first 72 hours of hospitalization to avoid interference in the responses about the child’s diet. Maternal factors investigated: age, schooling, income, parity, BMI and guidance on complementary feeding. Variables related to the child investigated: age, breastfeeding, infant school, BMI/age, height/age, weight/age and introduction of ultra-processed food. The association between the factors studied and introduction of ultra-processed food was tested by linear regression. The significance level considered was 0.05. Results . It was verified that only 21% of the children had not yet received any type of ultra-processed food, and 56.5% received any of these foods before 6 months. In the multivariate analysis, maternal schooling, family income, maternal age and parity were associated with ultra-processed food supply. Conclusions The feeding practices of children between 4 and 24 months are inadequate when compared to the recommendations for the age group.

The genus Echinococcus consists of parasites that have a life cycle with two mammalian hosts. Their larval stage, called the hydatid cyst, develops predominantly in the liver and lungs of intermediate hosts. The hydatid cyst is the causative agent of cystic hydatid disease and the species Echinococcus granulosus, G1 haplotype, is responsible for the vast majority of cases in humans, cattle and sheep. Protein characterization in hydatid cysts is essential for better understanding of the host-parasite relationship and the fertility process of Echinococcus. The aims of this work were the identification and quantitative comparison of proteins found in hydatid fluid from fertile and infertile cysts from E. granulosus, in order to highlight possible mechanisms involved in cyst fertility or infertility. Hydatid fluid samples containing proteins from both E. granulosus and Bos taurus were analysed by LC-MS/MS. Our proteomic analysis of fertile and infertile cysts allowed identification of a total of 498 proteins, of which 153 proteins were exclusively identified in the fertile cyst, 271 in the infertile cyst, and 74 in both. Functional in silico analysis allowed us to highlight some important aspects: (i) clues about the possible existence of an "arms race" involving parasite and host responses in fertile and infertile cysts; (ii) a number of proteins in hydatid fluid without functional annotation or with possible alternative functions; (iii) the presence of extracellular vesicles such as exosomes, which was confirmed by transmission electron microscopy.

Toxoplasma gondii is the causative agent of toxoplasmosis, an infectious disease that affects over 30% of the human world population, causing fatal infections in immunocompromised individuals and neonates. The life cycle of T. gondii is complex, and involves intermediate hosts (birds and mammals) and definitive hosts (felines, including domestic cats). The innate immune repertoire against the parasite involves the production of neutrophil extracellular traps (NET), and neutrophils from several intermediate hosts produce NET induced by T. gondii. However, the mechanisms underlying NET release in response to the parasite have been poorly explored. Therefore, the aims of this study were to investigate whether neutrophils from cats produce NET triggered by T. gondii and to understand the mechanisms thereby involved. Neutrophils from cats were stimulated with T. gondii tachyzoites and NET-derived DNA in the supernatant was quantified during the time. The presence of histone H1 and myeloperoxidase was detected by immunofluorescence. We observed that cat neutrophils produce both classical and rapid/early NET stimulated by T. gondii. Inhibition of elastase, intracellular calcium, and phosphatidylinositol 3-kinase (PI3K)-&delta, partially blocked classical NET release in response to the parasite. Electron microscopy revealed strands and networks of DNA in close contact or completely entrapping parasites. Live imaging showed that tachyzoites are killed by NET. We conclude that the production of NET is a conserved strategy to control infection by T. gondii amongst intermediate and definitive hosts.

The prion protein (PrPC) is predominantly expressed in the nervous and immune systems and is involved in relevant cell signaling. Microglia participate in neuroimmune interactions, and their regulatory mechanisms are critical for both health and disease. Despite recent reports with a microglial cell line, little is known about the relevance of PrPC in brain microglia. We investigated the role of PrPC in mouse primary microglia, and found no differences between wild type and Prnp-null cells in cell morphology or the expression of a microglial marker. Translocation of NF-κB to the nucleus also did not differ, nor did cytokine production. The levels of iNOS were also similar and, finally, microglia of either genotype showed no differences in either rates of phagocytosis or migration, even following activation. Thus, functional roles of PrPC in primary microglial cells are — if present — much more subtle than in transformed microglial cell lines.

Here we provide the LC–MS/MS data from a comparative analysis of Listeria monocytogenes ATCC 7644 treated and non-treated with a sublethal concentration of nisin (10 −3 mg/mL). Protein samples were analyzed by multidimensional protein identification technology (MudPIT) approach, in an off-line configuration. The raw MS/MS data allowed the detection of 49,591 spectra which resulted in 576 protein identifications. After Scaffold validation, 179 proteins were identified with high confidence. A label-free quantitative analysis based of normalized spectral abundance factor (NSAF) was used and 13 proteins were found differentially expressed between nisin-treated and non-treated cells. Gene ontology analysis of differentially expressed proteins revealed that most of them are correlated to metabolic process, oxidative stress response mechanisms and molecular binding. A detailed analysis and discussion of these data may be found in Miyamoto et al. [1] .

Echinococcus granulosus protoscolex proteins were separated using two-dimensional electrophoresis and then identified using mass spectrometry; we identified 61 proteins, 28 which are newly described of which 4 could be involved in hydatid cyst fertility molecular mechanisms.

Este trabalho se insere nos debates sobre a diversidade de significados dos esportes nas situações de lazer. Resulta de uma pesquisa etnográfica multilocalizada desenvolvida num grande circuito de ‘futebol varzeano’ da cidade de Porto Alegre. O objetivo foi compreender as dinâmicas de constituição das noções de violência nessa esfera de lazer, tendo como enfoque analítico dois ‘personagens empíricos’: o ‘guri’ e o ‘nego véio da várzea’. Esse empreendimento é efetivado num debate com a teoria elisiana em relação à violência e à busca da excitação agradável nas práticas miméticas do esporte. Ao final, apontamos dois aspectos como contribuições para os estudos do lazer: a existência de violências desagradáveis que não representam rupturas, mas que ‘fazem parte’ do contexto mimético, apontadas como desproporcionalidades; a vivência da tensão-excitação agradável não apenas num nível ótimo, mas também nos deslizamentos emocionais entre distintos equilíbrios de tensão. This work is part of the discussions on the diversity of sports meanings in leisure situations. It is the result of a multisite ethnographic research developed in a large circuit of ‘outskirts football’ in the city of Porto Alegre, Brazil. Its aim was to understand the dynamics of formation of the concepts of violence in leisure by focusing the analysis on two ‘empirical characters’: the ‘kid’ and the ‘old hand in outskirts football’. The enterprise is carried out within a debate with Eliasian theory on violence and the pursuit of pleasurable excitement in mimetic sporting practices. Finally, we point out two aspects as contributions to leisure studies: the existence of unpleasant types of violence that do not represent ruptures, but which ‘are part’ of that mimetic context, pointed out as disproportionalities; the experience of pleasant tension-excitement not only at an ideal level, but also in the emotional slidings between different tension balances. Este trabajo se inserta en los debates sobre a diversidad de significados de los deportes en las situaciones de ocio. Es resultado de una investigación etnográfica multilocalizada desarrollada en un gran circuito de ‘futbol de barrio’ de la ciudad de Porto Alegre. El objetivo fue comprender las dinámicas de constitución de las nociones de violencia en esa esfera de ocio, utilizando como foco analítico a dos ‘personajes empíricos’: el ‘chaval’ y el ‘pillo del barrio’. Ese trabajo é efectivado en un debate con la teoría elisiana en relación a la violencia y a la búsqueda de excitación agradable en las prácticas miméticas del deporte. Al final, señalamos como contribuciones: la existencia de violencias desagradables que no representan rupturas, sino que ‘forman parte’ del contexto mimético, apuntadas como desproporcionalidades; la vivencia de la tensiónexcitación agradable no sólo en un nivel óptimo, sino también en los deslizamientos emocionales entre distintos equilibrios de tensión.

Background In conditions of immunosuppression, the central nervous sty 5ystem (CNS) is the main target tissue for the reactivation of infection by Trypanosoma cruzi, the causative agent of Chagas disease. In experimental T. cruzi infection, interferon gamma (IFNγ)+ microglial cells surround astrocytes harboring amastigote parasites. In vitro, IFNγ fuels astrocyte infection by T. cruzi, and IFNγ-stimulated infected astrocytes are implicated as potential sources of tumor necrosis factor (TNF). Pro-inflammatory cytokines trigger behavioral alterations. In T. cruzi-infected mice, administration of anti-TNF antibody hampers depressive-like behavior. Herein, we investigated the effects of TNF on astrocyte susceptibility to T. cruzi infection and the regulation of cytokine production. Methods Primary astrocyte cultures of neonatal C57BL/6 and C3H/He mice and the human U-87 MG astrocyte lineage were infected with the Colombian T. cruzi strain. Cytokine production, particularly TNF, and TNF receptor 1 (TNFR1/p55) expression were analyzed. Recombinant cytokines (rIFNγ and rTNF), the anti-TNF antibody infliximab, and the TNFR1 modulator pentoxifylline were used to assess the in vitro effects of TNF on astrocyte susceptibility to T. cruzi infection. To investigate the role of TNF on CNS colonization by T. cruzi, infected mice were submitted to anti-TNF therapy. Results rTNF priming of mouse and human astrocytes enhanced parasite/astrocyte interaction (i.e., the percentage of astrocytes invaded by trypomastigote parasites and the number of intracellular parasite forms/astrocyte). Furthermore, T. cruzi infection drove astrocytes to a pro-inflammatory profile with TNF and interleukin-6 production, which was amplified by rTNF treatment. Adding rTNF prior to infection fueled parasite growth and trypomastigote egression, in parallel with increased TNFR1 expression. Importantly, pentoxifylline inhibited the TNF-induced increase in astrocyte susceptibility to T. cruzi invasion. In T. cruzi-infected mice, anti-TNF therapy reduced the number of amastigote nests in the brain. Conclusions Our data implicate TNF as a promoter of T. cruzi invasion of mouse and human astrocytes. Moreover, the TNF-enriched inflammatory milieu and enhanced TNFR1 expression may favor TNF signaling, astrocyte colonization by T. cruzi and egression of trypomastigotes. Therefore, in T. cruzi infection, a self-sustaining TNF-induced inflammatory circuit may perpetuate the parasite cycle in the CNS and ultimately promote cytokine-driven behavioral alterations. Electronic supplementary material The online version of this article (doi:10.1186/s12974-017-0952-0) contains supplementary material, which is available to authorized users.

Acanthamoeba polyphaga is a free-living protozoan pathogen, whose infective trophozoite form is capable of causing a blinding keratitis and fatal granulomatous encephalitis in humans. The damage caused by A. polyphaga trophozoites in human corneal or brain infections is the result of several different pathogenic mechanisms that have not yet been elucidated at the molecular level. We performed a comprehensive analysis of the proteins expressed by A. polyphaga trophozoites, based on complementary 2-DE MS/MS and gel-free LC–MS/MS approaches. Overall, 202 non-redundant proteins were identified. An A. polyphaga proteomic map in the pH range 3–10 was produced, with protein identification for 184 of 370 resolved spots, corresponding to 142 proteins. Additionally, 94 proteins were identified by gel-free LC–MS/MS. Functional classification revealed several proteins with potential importance for pathogen survival and infection of mammalian hosts, including surface proteins and proteins related to defense mechanisms. Our study provided the first comprehensive proteomic survey of the trophozoite infective stage of an Acanthamoeba species, and established foundations for prospective, comparative and functional studies of proteins involved in mechanisms of survival, development, and pathogenicity in A. polyphaga and other pathogenic amoebae.

Introduction. A systematic bias against women, resulting from the use of creatinine as a measure of renal function, has been identified in Model for End-stage Liver Disease (MELD)-based liver allocation. Correction of this bias by calculation of female creatinine levels using the Modification of Diet in Renal Disease (MDRD) formula has been suggested. Material and methods. A cohort of 639 cirrhotic candidates for firsttime liver transplantation was studied. Creatinine levels were corrected for gender using the MDRD formula. The accuracy of MELD, with or without creatinine correction, to predict 3- and 6-month mortality after inclusion in a transplant waiting list was estimated. Results. Women exhibited significantly lower creatinine levels, glomerular filtration rate, and MELD scores than men. After creatinine correction, female MELD scores had a mean increase of 1.1 points. Creatinine correction yielded an increase of 3 points in the MELD score in 15.2% of patients, 2 points in 22.4%, and 1 point in 17.6% of patients. The likelihood of death at 3 and 6 months after enrollment in the transplant waiting list was similar in males and females and the likelihood of receiving a transplant, as assessed by Kaplan-Meier survival curves, was also similar in males and females. Conclusion. The survival or the likelihood of receiving a transplant while on the waiting list were similar in men and women in both pre- and post-MELD eras and creatinine correction did not increase the accuracy of the MELD score in estimating 3- and 6-month mortality in female candidates for liver transplantation.

The hydatid fluid (HF) that fills Echinococcus multilocularis metacestode vesicles is a complex mixture of proteins from both parasite and host origin. Here, a LC-MS/MS approach was used to compare the HF composition of E. multilocularis H95 and G8065 isolates (EmH95 and EmG8065, respectively), which present differences in terms of growth and fertility. Overall, 446 unique proteins were identified, 392 of which (88%) were from parasite origin and 54 from culture medium. At least 256 of parasite proteins were sample exclusive, and 82 of the 136 shared proteins presented differential abundance between E. multilocularis isolates. The parasite's protein repertoires in EmH95 and EmG8065 HF samples presented qualitative and quantitative differences involving antigens, signaling proteins, proteolytic enzymes, protease inhibitors and chaperones, highlighting intraspecific singularities that could be correlated to biological features of each isolate. The repertoire of medium proteins found in the HF was also differential between isolates, and the relevance of the HF exogenous protein content for the parasite's biology is discussed. The repertoires of identified proteins also provided potential molecular markers for important biological features, such as parasite growth rate and fertility, as well potential protein targets for the development of novel diagnostic and treatment strategies for alveolar echinococcosis. Biological significance E. multilocularis metacestode infection of mammal hosts involve complex interactions mediated by excretory/secretory (ES) products. The hydatid fluid (HF) that fills the E. multilocularis metacestode vesicles contains complex repertoires of parasite ES products and host proteins that mediate important molecular interactions determinant for parasite survival and development, and, consequently, to the infection outcome. HF has been also extensively reported as the main source of proteins for the immunodiagnosis of echinococcosis. The performed proteomic analysis provided a comprehensive profiling of the HF protein composition of two E. multilocularis isolates. This allowed us to identify proteins of both parasite and exogenous (medium) origin, many of which present significant differential abundances between parasite isolates and may correlate to their differential biological features, including fertility and growth rate.

The objective of this study was to evaluate hormone and energy profiles in the postpartum period and to correlate these profiles with the resumption of ovarian cyclicity, as well as characterizing the postpartum short estrous cycle of Curraleiro Pé-Duro cows. Twelve Curraleiro Pé-Duro cows were examined via rectal palpation and ultrasound at 10 days postpartum, and subsequently examined daily to evaluate the resumption of ovarian cyclicity as well as every five days in order to evaluate uterine involution. Upon analysis of the data, it was possible to observe the formation of two distinct groups, one of which was comprised of those animals which returned to cyclicity within 60 days postpartum and another comprised of those animals which returned to cyclicity more than 105 days postpartum. Therefore, animals were divided into two groups; precocious, designated Ov Group, and delayed, designated NOv Group, wherein the cut-off time for all tests was 60 days postpartum. Statistically significant differences (P 0.01) between the groups occurred only regarding the day of 1st ovulation, which in the Ov Group averaged 51.4 ± 9.3 days and in the NOv Group averaged 138.3 ± 19.8 days postpartum. The other postpartum short estrous cycle variables assessed did not show statistically significant differences (P 0.05) between the groups. NEFA, BHBA and thyroxine concentration levels did not differ (P 0.05) between the groups in any of the statistical analyses. However, in the analysis comparing growth curves, triglycerides levels were higher for the Ov Group (P = 0.04) and cholesterol levels were higher for the NOv Group (P = 0.02). In this experiment, a small influence of a negative energy balance between the groups was observed, suggesting that these animals can present significant genetic variability due to natural selection, as evidenced by the formation of groups of animals with precocious and delayed reproductive characteristics.

Olfactory ensheathing cells (OECs) are a type of specialized glial cell currently considered as having a double function in the nervous system: one regenerative, and another immune. Streptococcus pneumoniae is a major agent of severe infections in humans, including meningitis. It is commonly found in the nasopharynx of asymptomatic carriers, and, under certain still unknown conditions, can invade the brain. We evaluated whether pneumococcal cells recovered from lysed OECs and microglia are able to survive by manipulating the host cell activation. An intracellular-survival assay of S. pneumoniae in OECs showed a significant number of bacterial CFU recovered after 3 h of infection. In contrast, microglia assays resulted in a reduced number of CFU. Electron-microscopy analysis revealed a large number of pneumococci with apparently intact morphology. However, microglia cells showed endocytic vesicles containing only bacterial cell debris. Infection of OEC cultures resulted in continuous NF-κB activation. The IFN-γ-induced increase of iNOS expression was reversed in infected OECs. OECs are susceptible to S. pneumoniae infection, which can suppress their cytotoxic mechanisms in order to survive. We suggest that, in contrast to microglia, OECs might serve as safe targets for pneumococci, providing a more stable environment for evasion of the immune system.

The objective of this study was to evaluate and compare the transfer of passive immunity and the proteinogram in Criollo Lageano (CL) and Black and White Holstein (BWH) calves. Two groups were utilized with 13 Criollo Lageano and 10 BWH calves. Blood samples were collected for the measurement of total serum protein, electrophoresis of serum proteins, activity of the gamma glutamyl transferase, and concentration of IgG by the method of the zinc sulfate turbidity in periods between 24 and 36 hours of life, 15, 30, 60, 90, 120, 150 and 180 days. Statistical analysis was performed by ANOVA and Tukey test at 5% significance level, and correlations between variables were calculated. Variations of serum proteins followed a pattern of physiological behavior over the first six months of life and production of immunoglobulins was active earlier in BWH calves and slower in the Criollo Lageano, without causing any impact on their health. Gamma globulin in the first days of life (24-36h) was correlated with IgG (r=0.87 for CL and r=0.89 for BWH), PTS (r=0.91 for CL and r=0.92 for BWH), Glob (r=0.99 for CL and r=0.98 for BWH) and GGT (r=0.14 for CL and r=0.83 for BWH). It was concluded that there was no failure in the transfer of passive immunity in Criollo Lageano calves but this failure occurred in the BWH calves. IgG values estimated by the zinc sulfate turbidity and serum proteins were considered good indicators of the transfer of passive immunity in calves between 24 and 36 hours of life. O objetivo do presente trabalho foi verificar e comparar a transferência de imunidade passiva e o proteinograma em bezerros da raça Crioula Lageana (CL) e Holandesa Preta e Branca (HPB). Foram utilizados dois grupos sendo 13 bezerros da raça Crioula Lageana e 10 da raça HPB. Amostras de sangue foram colhidas para a mensuração da proteína sérica total, eletroforese das proteínas séricas, atividade da gama-glutamiltransferase e concentração de IgG pelo método de turvação pelo sulfato de zinco nos períodos entre 24 e 36 horas de vida, 15, 30, 60, 90, 120, 150 e 180 dias. A análise estatística foi realizada pelo teste de ANOVA e Tukey ao nível de 5% de significância, e correlações entre as variáveis. As variações das proteínas séricas obedeceram a um padrão de comportamento fisiológico ao longo dos seis primeiros meses de vida e a produção ativa de imunoglobulinas foi mais precoce nos bezerros da raça Holandesa e mais lenta nos da raça Crioula Lageana, sem provocar qualquer impacto sobre a saúde dos mesmos. A gamaglobulina no primeiro dia de vida (24-36h) correlacionou-se com a IgG (r=0,87 CL e r=0,89 HPB), PTS (r=0,91 CL e r=0,92 HPB), Glob (r=0,99 CL e r=0,98) e GGT (r=0,14 CL e r=0,83 HPB). Conclui-se que não houve falha na transferência de imunidade passiva nos bezerros da raça Crioula Lageana e falha na raça Holandesa Preta e Branca. Os valores de IgG estimados pelo método da turvação pelo sulfato de zinco e das proteínas séricas foram bons indicadores da transferência de imunidade passiva nos bezerros entre 24 e 36 horas de vida.

Cystic hydatid disease (CHD) is caused by infection with Echinococcus granulosus metacestodes and affects humans and livestock. Proteins secreted or excreted by protoscoleces, pre-adult worms found in the metacestode, are thought to play fundamental roles in the host-parasite relationship. In this work, we performed an LC-MS/MS proteomic analysis of the excretory-secretory products obtained from the first 48 h of an in vitro culture of the protoscoleces. We identified 32 proteins, including 18 that were never detected previously in metacestode proteomic studies. Among the novel identified excretory-secretory products are antigenic proteins, such as EG19 and P-29 and a calpain protease. We also identified other important protoscolex excretory-secretory products, such as thioredoxin peroxidase and 14-3-3 proteins, which are potentially involved in evasion mechanisms adopted by parasites to establish infection. Several intracellular proteins were found in the excretory-secretory products, revealing a set of identified proteins not previously thought to be exposed at the host-parasite interface. Additionally, immunological analyses established the antigenic profiles of the newly identified excretory-secretory products and revealed, for the first time, the in vitro secretion of the B antigen by protoscoleces. Considering that the excretory-secretory products obtained in vitro might reflect the products released and exposed to the host in vivo, our results provide valuable information on parasite survival strategies in adverse host environments and on the molecular mechanisms underpinning CHD immunopathology.

Echinaster (Othilia) guyanensis Clark, 1987 is an endangered starfish distributed throughout the Caribbean and Atlantic Ocean. Even though it has been extensively harvested, little is known about the biology and ecology of this starfish. Here, we examine reproduction seasonality in E. (O.) guyanensis. Individuals were collected monthly for one year, including four complete lunar phases. The gonad index (GI) was calculated to determine annual and monthly reproductive peaks. Gametogenesis stages were also determined. Sex ratio was 1:1.33 (M:F). Gonadosomatic index, body weight, central disc width and arm length were similar for both sexes. Gonads were present in all animals with arm length greater than 36.2 mm. Lunar phase was not associated with E. (O.) guyanensis reproduction. GI and gametogenesis patterns suggest that starfish have an annual reproductive peak with spawning during autumn months (March to May).

Alcohol use is highly prevalent in patients with bipolar disorder (BD) and is associated with significant mortality and morbidity. The detrimental effects of each condition are compounded by the presence of the other. The objective of this study was to examine the impact of alcohol abuse and of alcohol dependence in BD in a Brazilian sample, as indicated by clinical severity, functional impairment, and quality of life (QOL). A cross-sectional survey of 186 bipolar outpatients were interviewed using the Structured Clinical Interview for the Diagnostic and Statistical Manual of Mental Disorders-4th Edition. The primary outcome measures were functioning, as indicated by the Global Assessment of Functioning Scale scores and QOL, as indicated by the World Health Organization Quality of Life Instrument. Secondary outcomes were clinical severity features. Alcohol abuse and dependence were associated with male gender, lower education, earlier age of onset, psychosis within first episode, depressive symptoms, and worse functioning. In addition, the presence of alcohol abuse or dependence was associated with remarkably high rates of suicide attempt. Our findings suggest that the co-occurrence of alcohol abuse/dependence with BD increases the risk for suicide attempt, which may reflect in part the greater severity of symptoms and impaired functioning. This subgroup of bipolar patients requires a treatment tailored to address both conditions.

Accumulating evidences have assigned a central role to parasite-derived proteins in immunomodulation. Here, we report on the proteomic identification and characterization of immunomodulatory excretory-secretory (ES) products from the metacestode larva (tetrathyridium) of the tapeworm Mesocestoides corti (syn. M. vogae). We demonstrate that ES products but not larval homogenates inhibit the stimuli-driven release of the pro-inflammatory, Th1-inducing cytokine IL-12p70 by murine bone marrow-derived dendritic cells (BMDCs). Within the ES fraction, we biochemically narrowed down the immunosuppressive activity to glycoproteins since active components were lipid-free, but sensitive to heat- and carbohydrate-treatment. Finally, using bioassay-guided chromatographic analyses assisted by comparative proteomics of active and inactive fractions of the ES products, we defined a comprehensive list of candidate proteins released by M. corti tetrathyridia as potential suppressors of DC functions. Our study provides a comprehensive library of somatic and ES products and highlight some candidate parasite factors that might drive the subversion of DC functions to facilitate the persistence of M. corti tetrathyridia in their hosts., Author Summary The metacestode larval stages of life-threatening tapeworms grow within the organs of its mammalian hosts, thus causing severe and long-lasting morbidity. Immunosuppression, which mainly depends on factors that are released or leaking from the parasite, plays an important role in both survival and proliferation of the larvae. These parasite-derived molecules are potential targets for developing new anti-parasitic drugs and/or improving the effectiveness of current therapies. Moreover, an optimized use of such factors could help to minimize pathologies resulting from uncontrolled immune responses, like allergies and autoimmune diseases. The authors herein demonstrate that larvae from a parasitic cestode release factors that sufficiently support the suppression of dendritic cells, a set of innate immune cells that recognizes and initiates host immune responses against invading pathogens. Employing modern analytic proteomic tools combined with immunological bioassays, several cestode-derived candidate immunomodulators were identified. This is the first bioassay-guided comprehensive library of candidate immunomodulators from a tissue-dwelling cestode larva. This work validates the unmet value of the Mesocestoides corti system in characterizing the mechanisms of host immunomodulation by metacestodes and reveals the largest database of candidate metacestode-derived immunomodulators until date.

Background: Ultraviolet B (UVB) radiation can cause morphological and biological alterations similar to those of melanoma in situ in irradiated melanocytic nevi. matrix metalloproteinase (MMP-2) and -9 appear to be involved with tumour invasion, the formation of metastases and neoangiogenesis in melanomas. The effects of UVB on the immunohistochemical expression of gelatinases in different cell types in melanocytic nevi have not been completely studied. Purpose: Evaluate the effects of UVB on the immunohistochemical expression of MMP-2 and -9 in different cell types in melanocytic nevi. Methods: Forty-two melanocytic nevi had one half irradiated with 2 MEDs of UVB and were excised 1 week later. Three different observers compared the intensity of the immunohistochemical expression of gelatinases in keratinocytes, melanocytes, endothelial cells and fibroblasts on the irradiated (IS) and non-irradiated sides (NIS). Results: All the cell types showed an increase in the expression of MMP-2 on the IS, especially the epidermal melanocytes. No significant increase in the expression of MMP-9 in keratinocytes was detected on the IS; significant increase was observed in all the remaining cells. Conclusions: One single irradiation of UVB increases the immunohistochemical expression of gelatinases in almost all evaluated cell lines, with the exception of MMP-9 in keratinocytes.

Este foi um estudo prospectivo que visou identificar a microbiologia do meato médio em pacientes com rinossinusite crônica (RSC) e compará-la com a de indivíduos sadios. MATERIAL E MÉTODOS: Foram incluídos 134 pacientes RSC e 50 voluntários sadios, que constituíram o grupo controle. As amostras foram coletadas endoscopicamente e submetidas a exames pelo método de Gram com contagem leucocitária e culturas para aeróbios, anaeróbios e fungos. RESULTADOS: Nos pacientes com RSC foram cultivados 220 microorganismos, dentre os quais os mais freqüentes foram o Staphylococcus aureus, presente em 31% das amostras, e o Staphylococcus coagulase-negativo (SCN) em 23%. Gram-negativos ou facultativos foram isolados em 37% das amostras, anaeróbios em 12%, e fungos em 14%. Ao exame bacterioscópico evidenciou-se alguns ou numerosos leucócitos em 74% das amostras com culturas positivas. Nos indivíduos sadios o SCN foi isolado em 40% das amostras e o Staphylococcus aureus em 18%. Em 12% dos indivíduos a cultura para fungos foi positiva, e o exame direto negativo. Todas as culturas anaeróbias foram estéreis. Quanto à contagem leucocitária todos apresentaram nenhum ou raros leucócitos. CONCLUSÃO: Os grupos apresentaram resultados semelhantes quanto à microbiologia, entretanto, diferiram em relação à contagem leucocitária, o que auxilia na diferenciação um microorganismo infectante de um colonizante. This was a prospective study which assessed endoscopically collected middle meatus secretions in patients with chronic rhinosinusitis (CRS) and compared those findings with microbiological data of healthy individuals. METHODS: Middle meatus samples were collected from 134 CRS patients. In the laboratory, samples were Gram stained for microscopic examination with white blood cels (WBCs) count and also send for aerobic, anaerobic and fungal cultures. Fifty volunteers served as control. RESULTS: In CRS patients a total of 220 microorganisms were isolated. The most frequent microorganisms were Staphylococcus aureus (31%), coagulase-negative Staphylococcus (CNS) (23%). Gram-negative or facultative microorganisms were isolated in 37% of the samples, anaerobes in 12% and fungi in 14%. Seventy four percent of the samples with positive cultures presented many or few WBC. In the control group, 76% of cultures were positive for aerobes and 12% for fungi. No anaerobes were isolated. There were rare or no WBC in the fifty samples. The most frequent microorganisms were CNS (40%), Staphylococcus aureus (18%). CONCLUSION: The microbiology of the middle meatus is similar in CRS patients and healthy individuals. Despite this, there was an important difference between the WBC count in these two groups, which helps to distinguish an infective from a saprophitic microorganism.

Tritrichomonas foetus is an amitochondriate parasite that possesses hydrogenosomes, unusual anerobic energy-producing organelles. In these organisms the "mitochondrial cell death machinery" is supposed to be absent, and the mechanisms that lead to cell demise remain to be elucidated. The presence of a cell death program in trichomonads has already been reported, suggesting the existence of a caspase-like execution pathway in such organisms. Here we demonstrate the alterations provoked by the fungicide griseofulvin and raise the possibility that other cell death pathways may exist in T. foetus. Dramatic changes in trichomonads morphology are presented after griseofulvin treatment, such as intense plasma membrane and nuclear envelope blebbing, nucleus fragmentation, and an abnormal number of oversized vacuoles. One important finding was the exposition of phosphatidylserine (PS) in the outer leaflet of the plasma membrane in cells after drug treatment, and also the presence of a high amount of misshapen flagella and tubulin precipitates as vacuolar contents, suggesting an autophagic process of abnormal cellular elements. Interestingly, immunoreactivity for activated caspase-3 was not detected during griseofulvin treatment, a finding distinct from the observed when this cell was treated with H(2)O(2). The possibility of the existence of different pathways to cell death in trichomonads is discussed.

BACKGROUND: Patients infected with the human immunodeficiency virus (HIV) have generally been excluded from consideration for liver transplantation. Recent advances in the management and prognosis of these patients suggest that this policy must be reevaluated. AIM: To identify the current position of Brazilian transplant centers concerning liver transplantation in asymptomatic HIV-infected patients with end-stage liver disease. METHODS: A structured questionnaire was submitted by e-mail to Brazilian groups who perform liver transplantation and were active in late 2003, according to the Brazilian Association of Organ Transplantation. RESULTS: Of the 53 active groups, 30 e-mail addresses have been found of professionals working in 41 of these groups. Twenty-one responses (70%) were obtained. Most of the professionals (62%) reported that they do not include HIV-infected patients in waiting lists for transplants, primarily on account of the limited world experience. They also reported, however, that this issue will soon be discussed by the group. Those who accept these patients usually follow the guidelines provided by the literature: patients must fulfill the same inclusion criteria as the other patients with end-stage liver diseases, present low or undetectable HIV viral load, and a CD4 count above 250/mm³. They reported that there are 10 HIV-infected patients in waiting list and that only one patient has received a liver transplant in the country. CONCLUSION: Most centers do not accept in waiting lists for liver transplantation patients with HIV infection, even asymptomatic ones. However, advances in the management of HIV-infected patients suggest that this policy must be reevaluated. In Brazil, there is practically no experience in liver transplantation in HIV-positive patients.

Giardia lamblia is the most commonly detected parasite in the intestinal tract of humans and other mammals causing giardiasis. Giardia presents several cytoskeletal structures with microtubules as major components such as the ventral adhesive disk, eight flagella axonemes, the median body and funis. Many drugs have already been tested as antigiardial agents, such as albendazole and mebendazole, which act by specifically inhibiting tubulin polymerization and hence microtubule assembly. In the present work, we used the microtubule inhibitors nocodazole and colchicine in order to investigate their direct and indirect effects on Giardia ultrastructure and attachment to the glass surface, respectively. Axenically grown G. lamblia trophozoites were treated with nocodazole or colchicine for different time intervals and analyzed by light and electron microscopy. It was observed that trophozoites became completely misshapen, detached from the glass surface and failed to complete cell division. The main alterations observed included disc fragmentation, presence of large vacuoles, and appearance of electrondense deposits made of tubulin. The cytokinesis was blocked, but not the karyokinesis, and membrane blebs were observed. These findings show that Giardia behavior and cytoskeleton are clearly affected by the commonly used microtubule targetting agents colchicine and nozodazole.

Tritrichomonas foetus is a parasitic protist of the urogenital tract of cattle. It presents the trophozoite stage, a motile elongated form that constitutes most of the cells in a normal population, and a pseudocyst stage, an immotile rounded form that appears under unfavourable environmental conditions. In the present report pseudocysts were studied in natural conditions and after induction by chemicals or cycles of cooling and warming of cultures. The capacity of T. foetus to adhere to vaginal epithelial cells (VECs) was compared for both trophozoite and pseudocyst forms. By the use of video-enhanced-contrast microscopy, scanning and transmission electron microscopy, and immunofluorescence microscopy techniques, we present evidence that: (1) T. foetus easily internalizes the flagella and forms pseudocysts under several unfavourable conditions; (2) T. foetus in both pseudocyst and trophozoite forms is able to adhere to VECs; (3) the adhesion rate is higher for pseudocysts than for trophozoites; (4) the adhesin Tf190 is expressed in both forms during interaction; (5) the adhesion process of pseudocysts seems to occur in a contact-dependent manner. Thus, we propose that the pseudocyst stage is not a degenerative form, but a functional life form that is able to interact with and firmly adhere to VECs.

We present observations on an unusual tetratrichomonad species isolated from preputial smegma of virgin bulls. Ultrastructural studies were performed using scanning and electron microscopy techniques. This protozoan presents four anterior flagella of unequal length and a recurrent one forming the undulating membrane. It shows one anterior nucleus, a Golgi complex, an axostyle, and a costa. The hydrogenosomes are rather elongated, seen in groups, and presenting different electron densities. Vacuoles of different sizes containing bacteria and material in process of digestion were frequently found. PCR was also used in order to compare the species herein described with other trichomonad species. The amplification products were seen only with primers TFR1 and TFR2 (specific to trichomonads), but not with TFR3 and TFR4 (specific to Tritrichomonas foetus), suggesting that although collected from the genital tract of the bull, this protist was not T. foetus. We propose that the appearance of these tetratrichomonads were probably due to the sodomy practiced among bulls. Concomitant contamination of preputial cavity with feces could explain the presence of the opportunistic organism. The observations presented here show the importance of the correct diagnostic when investigating samples obtained from the urogenital tract of cattle. We also suggest that this flagellate belongs to the species Tetratrichomonas buttreyi.

Tritrichomonas foetus is an amitochondrial parasite protist which lacks typical eukaryote organelles such as mitochondria and peroxisomes, but possesses the hydrogenosome, a double-membrane-bound organelle that produces ATP. The cell death of amitochondrial organisms is poorly studied. In the present work, the cytotoxic effects of hydrogen peroxide on T. foetus and its participation on cell death were analyzed. We took advantage of several microscopy techniques, including videomicroscopy, light microscopy immunocytochemistry for detection of caspase activation, and scanning and transmission electron microscopy. We report here that in T. foetus: (1) H(2)O(2) leads to loss of motility and induces cell death, (2) the dying cells exhibit some characteristics similar to those found during the death of other organisms, and (3) a caspase-like protein seems to be activated during the death process. Thus, we propose that, although T. foetus does not present mitochondria nor any known pathways of cell death, it is likely that it bears mechanisms of cell demise. T. foetus exhibits morphological and physiological alterations in response to H(2)O(2) treatment. The hydrogenosome, a unique organelle which is supposed to share a common ancestral origin with mitochondria and has an important role in oxidative responses in trichomonads, is a candidate for participating in this event.

OBJECTIVE: To assess survival of patients undergoing cerebral cardiopulmonary resuscitation maneuvers and to identify prognostic factors for short-term survival. METHODS: Prospective study with patients undergoing cardiopulmonary resuscitation maneuvers. RESULTS: The study included 150 patients. Spontaneous circulation was re-established in 88 (58%) patients, and 42 (28%) were discharged from the hospital. The necessary number of patients treated to save 1 life in 12 months was 3.4. The presence of ventricular fibrillation or tachycardia (VF/VT) as the initial rhythm, shorter times of cardiopulmonary resuscitation maneuvers and cardiopulmonary arrest, and greater values of mean blood pressure (BP) prior to cardiopulmonary arrest were independent variables for re-establishment of spontaneous circulation and hospital discharge. The odds ratios for hospital discharge were as follows: 6.1 (95% confidence interval [CI] = 2.7-13.6), when the initial rhythm was VF/VT; 9.4 (95% CI = 4.1-21.3), when the time of cerebral cardiopulmonary resuscitation was < 15 min; 9.2 (95% CI = 3.9-21.3), when the time of cardiopulmonary arrest was < 20 min; and 5.7 (95% CI = 2.4-13.7), when BP was > 70 mmHg. CONCLUSION: The presence of VF/VT as the initial rhythm, shorter times of cerebral cardiopulmonary resuscitation and of cardiopulmonary arrest, and a greater value of BP prior to cardiopulmonary arrest were independent variables of better prognosis.

We present observations on the fine structure and the division process of the nucleus in the protist Tritrichomonas foetus, parasite of the urogenital tract of cattle. The nucleus was followed by immunofluorescence and electron microscopy during interphase and mitosis. Conventional karyotyping coupled to image processing and bright field Panotic staining were used to follow nucleus modifications, chromosome number and condensation pattern along the whole cell cycle. Confocal laser scanning microscopy (CLSM) using DNA fluorescent probes, followed by image processing in the SURF-Driver program, produced three-dimensional reconstruction data of the mitotic nucleus under each phase of the division process. Immunocytochemistry in thin-sections revealed the chromosome spatial arrangement after bromodeoxyuridine incorporation and immunogold labeling using anti-DNA monoclonal antibodies. Our results indicate that: (1) the nucleus assumes different size and shapes along mitosis: it appears oval in interphase, becoming lobed or concave in prophase, then undergoing torsion and constriction, displaying an 'S' shape (metaphase). Next, it becomes elongated and it is finally separated in two nuclei at the transition of anaphase to telophase; (2) T. foetus nucleus harbors five chromosomes; (3) chromosomes become condensed in a pre-mitotic phase; (4) the nucleolus persists during the mitosis.

Listeria monocytogenes infections have been frequently reported in many food poisoning outbreaks around the world. In this work, the protein repertoires of L. monocytogenes ATCC 7644 cells treated or not with a 10− 3 mg/mL nisin sublethal concentration, established by antimicrobial susceptibility tests, were analyzed by LC–MS/MS. Overall, 179 proteins were identified, 9 of them more abundant in nisin-treated samples, and 4 more abundant in non-treated control samples. In nisin treated cells, proteins associated to oxidative stress response showed higher abundance. Also, the higher abundance of an enzyme related to the production of cell membrane lipids upon nisin exposure is suggestive of both a failure in conventional cell division mechanism and the activation of an alternative L-form mediated division mechanism. Finally, flagellar and motility proteins' overexpression upon nisin exposure is indicative of increased bacterial motility in response to the bacteriocin. Taken together, these results provide new insights on nisin effects on L. monocytogenes cells and on how this bacterium may overcome a bacteriocin-containing environment. Biological significance The antimicrobial mechanism of nisin on target bacterial cells has been extensively studied since discovery of this bacteriocin. The nisin pore-forming mechanism is mediated by its binding to the pyrophosphate portion of membrane lipid II [1] , but some evidences point out to alternative mechanisms. Results from assays with mutacin 1140 hybrids [2] showed that the portion of nisin that is not involved with lipid II binding could damage the bacterial cell, independently of pore formation [3] , [4] . Moreover, there are insufficient data to explain how nisin affects the bacterial survival. In this scenario, proteomics is an interesting approach, as a comparison between treated and untreated cells may provide insights of both antimicrobial mechanisms of action and bacterial response mechanisms [5] .

Summary With the aim of starting a programme to conserve the Wild Horse of Roraima and the Northeastern Donkey, we adapted techniques used on bovine embryos to these equids. During the breeding season of 1987/88, 120 embryo collection attempts were performed on 11 crossbred donor mares between Days 6 and 8 after ovulation. Most attempts (107) were carried out on Day 6 to give an overall recovery rate of 48.3 per cent. Of 20 Day 6 embryos that were frozen in liquid nitrogen and subsequently thawed, 17 (85 per cent) appeared viable and were transferred non-surgically to recipient mares synchronised to within +1 to −5 days of the embryo donors. This resulted in four pregnancies at Day 60 (23.5 per cent) compared to three pregnancies resulting from the transfer of seven ‘fresh’ unfrozen embryos (43 per cent). Transfer of 10 bisected demi-embryos to individual recipients resulted in one live foal.

The objective of this study was to use morphometric and ultrasound evaluations to estimate the growth curve of the Pantaneiro cattle breed, raised in its natural habitat, aiming at the re-insertion of this breed in production systems. One hundred and three females, aging from months to 11 years, and raised on native pastures, were evaluated. The animals belonged to the Conservation Nucleus of Embrapa Pantanal, located in the Pantanal of Mato Grosso do Sul (Brazil). Weight, thoracic perimeter (TP), body length (BL), rump height (RH), height at withers (HW), hip height (HH), depth (DP), distance between the ilia (DI) (cm) and rib-eye area (REA) were measured. To relate the measurements with the age of the animals, the univariate regression model was used, assigning the variable response to gamma distribution. The Pearson correlation between variables was also estimated. The inflection point of the growth curve was 37 months for HH; between 38 and 39 months for TP and HW; between 40 and 41 months for DI, HH and DP; and 45 months for BL. The REA results could not fit in a statistical model. The majority of the variables presented a correlation above 60% among themselves, except for REA × Age, of 15.81%; REA × HW, of 34.44%; HH × Age, of 46.19; HH × DI, of 58.07%; REA × HH, of 24.57%; and REA × TP, of 39.9%. The cows showed maturity age at 40 months, which may have occurred because they were raised in natural farming conditions. In Pantaneiro cows reared in extensive systems only on natural pastures, the use of ultrasound is not effective to estimate the curve of muscular development, perhaps because this breed was not selected for weight gain.

Hemocytes are the first line of defense of the immune system in invertebrates, but despite their important role and enormous potential for the study of gene-environment relationships, research has been impeded by a lack of consensus on their classification. Here we used flow cytometry combined with histological procedures, histochemical reactions and transmission electron microscopy to characterize the hemocytes from the oyster Crassostrea rhizophorae. Transmission electron microscopy revealed remarkable morphological characteristics, such as the presence of membranous cisternae in all mature cells, regardless of size and granulation. Some granular cells contained many cytoplasmic granules that communicated with each other through a network of channels, a feature never previously described for hemocytes. The positive reactions for esterase and acid phosphatase also indicated the presence of mature cells of all sizes and granule contents. Flow cytometry revealed a clear separation in complexity between agranular and granular populations, which could not be differentiated by size, with cells ranging from 2.5 to 25 µm. Based on this evidence we suggest that, at least in C. rhizophorae, the different subpopulations of hemocytes may in reality be different stages of one type of cell, which accumulates granules and loses complexity (with no reduction in size) as it degranulates in the event of an environmental challenge.

The prion protein (PrP(C)) is a cell surface protein expressed mainly in the nervous system. In addition to the role of its abnormal conformer in transmissible spongiform encephalopathies, normal PrP(C) may be implicated in other degenerative conditions often associated with inflammation. PrP(C) is also present in cells of hematopoietic origin, including T cells, dendritic cells, and macrophages, and it has been shown to modulate their functions. Here, we investigated the impact of inflammation and stress on the expression and function of PrP(C) in neutrophils, a cell type critically involved in both acute and chronic inflammation. We found that systemic injection of LPS induced transcription and translation of PrP(C) in mouse neutrophils. Up-regulation of PrP(C) was dependent on the serum content of TGF-β and glucocorticoids (GC), which, in turn, are contingent on the activation of the hypothalamic-pituitary-adrenal axis in response to systemic inflammation. GC and TGF-β, either alone or in combination, directly up-regulated PrP(C) in neutrophils, and accordingly, the blockade of GC receptors in vivo curtailed the LPS-induced increase in the content of PrP(C). Moreover, GC also mediated up-regulation of PrP(C) in neutrophils following noninflammatory restraint stress. Finally, neutrophils with up-regulated PrP(C) presented enhanced peroxide-dependent cytotoxicity to endothelial cells. The data demonstrate a novel interplay of the nervous, endocrine, and immune systems upon both the expression and function of PrP(C) in neutrophils, which may have a broad impact upon the physiology and pathology of various organs and systems.

Background Antigen B (AgB) is the major protein secreted by the Echinococcus granulosus metacestode and is involved in key host-parasite interactions during infection. The full comprehension of AgB functions depends on the elucidation of several structural aspects that remain unknown, such as its subunit composition and oligomeric states. Methodology/Principal Findings The subunit composition of E. granulosus AgB oligomers from individual bovine and human cysts was assessed by mass spectrometry associated with electrophoretic analysis. AgB8/1, AgB8/2, AgB8/3 and AgB8/4 subunits were identified in all samples analyzed, and an AgB8/2 variant (AgB8/2v8) was found in one bovine sample. The exponentially modified protein abundance index (emPAI) was used to estimate the relative abundance of the AgB subunits, revealing that AgB8/1 subunit was relatively overrepresented in all samples. The abundance of AgB8/3 subunit varied between bovine and human cysts. The oligomeric states formed by E. granulosus AgB and recombinant subunits available, rAgB8/1, rAgB8/2 and rAgB8/3, were characterized by native PAGE, light scattering and microscopy. Recombinant subunits showed markedly distinct oligomerization behaviors, forming oligomers with a maximum size relation of rAgB8/3>rAgB8/2>rAgB8/1. Moreover, the oligomeric states formed by rAgB8/3 subunit were more similar to those observed for AgB purified from hydatid fluid. Pressure-induced dissociation experiments demonstrated that the molecular assemblies formed by the more aggregative subunits, rAgB8/2 and rAgB8/3, also display higher structural stability. Conclusions/Significance For the first time, AgB subunit composition was analyzed in samples from single hydatid cysts, revealing qualitative and quantitative differences between samples. We showed that AgB oligomers are formed by different subunits, which have distinct abundances and oligomerization properties. Overall, our findings have significantly contributed to increase the current knowledge on AgB expression and structure, highlighting issues that may help to understand the parasite adaptive response during chronic infection., Author Summary Antigen B (AgB) is the major secretory protein of the Echinococcus granulosus hydatid cyst, the causative agent of cystic hydatid disease. Structurally, AgB is a multisubunit protein formed by 8-kDa subunits, but it is not known which subunits are secreted by a single parasite (cyst) and how they interact in the formation of distinct AgB oligomeric states. Here, we investigated AgB subunit composition and oligomeric states in individual samples from bovine and human cysts. We identified AgB8/1, AgB8/2, AgB8/3 and AgB8/4 subunits in AgB oligomers of all samples analyzed. Quantitative and qualitative differences in the expression of AgB subunits were observed within and between samples. Using recombinant subunits as models, we showed that AgB subunits form distinct oligomeric states, with a rAgB8/3>rAgB8/2>rAgB8/1 maximum size relation. We also demonstrated by different experimental approaches that rAgB8/3 oligomers are more similar, both in size and morphology, to those observed for E. granulosus AgB. Overall, we provided experimental evidences that AgB is composed of different subunits within a single cyst, and that subunits have different abundances and oligomerization properties. These issues are important for the understanding of AgB expression and structure variations, and their impact for the host-parasite cross-talk.

Renal tubular cells are the main target of ischemic insult associated with acute renal injury. Low oxygen and nutrient supplies result in ATP depletion, leading to cell death and loss of renal function. A possible mechanism by which bone marrow-derived cells support renal tissue regeneration relies on the capacity of mononuclear cells (BMMC), particularly mesenchymal stem cells (MSC), to secrete paracrine factors that mediate support for kidney regeneration.BMMC/MSC and renal cells (LLC-PK(1) from pig and IRPTC from rat) were co-cultured under stressful conditions (ATP depletion and/or serum free starvation), physically separated by a microporous membrane (0.4 μm), was used to determine whether bone marrow-derived cells can interact with renal cells in a paracrine manner.This interaction resulted in stimulation of renal cell proliferation and the arrest of cell death. MSC elicit effective responses in renal cells in terms of stimulating proliferation and protection. Such effects are observed in renal cells co-cultured with rat BMMC/MSC, an indication that paracrine mechanisms are not entirely species-specific.The paracrine action of BMMC/MSC was influenced by a renal cell stimulus released during stress, indicating that cross-talk with injured cells is required for renal regeneration supported by bone marrow-derived cells.