Unravelling the role of 17ß-estradiol on advancing uterine luteolytic cascade in cattle.

In cattle, 17ß-estradiol (E2) stimulates prostaglandin F2α (PGF2α) synthesis, which causes luteolysis. Except for the well-established upregulation of oxytocin receptor gene (OXTR), molecular mechanisms of E2-induced PGF2α release in vivo remain unknown. We hypothesized that E2-induced PGF2α release requires de novo transcription of components of the PGF2α synthesis machinery. Beef cows (n = 52) were assigned to remain untreated (Control; n = 10), to receive 50% ethanol infusion intravenously (Placebo; n = 21), or 3 mg E2 in 50% ethanol infusion intravenously (Estradiol; n = 21) on day 15 (D15) after estrus. We collected a single endometrial biopsy per animal at the time of the treatment (0h; Control B0h group), 4 hours (4h; Placebo B4h group and Estradiol B4h group), or 7 hours (7h; Placebo B7h group and Estradiol B7h group) post-treatment. Compared to the Placebo group, the Estradiol group presented significantly greater 13,14-dihydro-15-keto-PGF2α concentrations between 4h and 7h and underwent earlier luteolysis. At 4h, the qPCR analysis showed a lower abundance of ESR1, ESR2 and aldo-keto reductase family 1 member B1 (AKR1B1) genes in the Estradiol B4h group, and a greater abundance of OXTR compared to the Placebo B4h group. Similarly, the E2 treatment significantly reduced the abundance of AKR1B1, and AKR1C4 in the Estradiol B7h group, compared to the placebo group. Overall, E2-induced PGF2α release and luteolysis involved an unexpected and transient downregulation of components of the PGF2α-synthesis cascade, except for OXTR, which was upregulated. Collectively, our data suggest that E2 connects newly-synthesized OXTR to pre-existing cellular machinery to synthesize PGF2α and cause luteal regression.

Lung injury caused by occupational exposure to particles from the industrial combustion of cashew nut shells: a mice model.

Cashew nut shells (CNS) is already used in the energy matrix of some industries. However, it is necessary to know the harmful health effects generated by exposure to pollutants of its combustion, especially in the workers exposed to industrial pollutants. In addition, it is known that the incidence of asthma grows among workers in industries, and due to its previously reported biological effects of anethole, these will also be objects of the present study. We used 64 Balb/C mice, randomly divided into eight groups. Groups were sensitized and challenged with saline or ovalbumin, then subjected to intranasal instillation of 30 µg PM4.0 (occupational exposure) from the combustion of CNS or saline, and then were subsequently treated with oral anethole 300 mg/kg or 0.1% Tween 80. Our results serve as a starting point for the development of public policies for the prevention of diseases in workers that are exposed to the pollutants coming from industries.

PRL3 enhances T-cell acute lymphoblastic leukemia growth through suppressing T-cell signaling pathways and apoptosis.

T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive malignancy of thymocytes and is largely driven by the NOTCH/MYC pathway. Yet, additional oncogenic drivers are required for transformation. Here, we identify protein tyrosine phosphatase type 4 A3 (PRL3) as a collaborating oncogenic driver in T-ALL. PRL3 is expressed in a large fraction of primary human T-ALLs and is commonly co-amplified with MYC. PRL3 also synergized with MYC to initiate early-onset ALL in transgenic zebrafish and was required for human T-ALL growth and maintenance. Mass-spectrometry phosphoproteomic analysis and mechanistic studies uncovered that PRL3 suppresses downstream T-cell phosphorylation signaling pathways, including those modulated by VAV1, and subsequently suppresses apoptosis in leukemia cells. Taken together, our studies have identified new roles for PRL3 as a collaborating oncogenic driver in human T-ALL and suggest that therapeutic targeting of the PRL3 phosphatase will likely be a useful treatment strategy for T-ALL.

Supplemental progesterone induces temporal changes in luteal development and endometrial transcription in beef cattle.

Long-acting injectable progesterone (iP4) supplementation during early diestrus is a strategy to increase conception rates in cow-calf beef operations. However, iP4 treatment causes early functional and structural regression of the corpus luteum (CL) in a proportion of iP4-treated animals, resulting in pregnancy loss. The hypothesis evaluated was that iP4 accelerates downregulation of sex-steroid receptors (PGR, ESR1, ESR2) during early to mid-diestrus and the upregulation of genes controlling PGF2α secretion (OXTR, PTGS2, AKR1B1) during late diestrus in the endometrium. Ovulations of cyclic, multiparous Nelore (Bos indicus) cows were synchronized, and cows were divided to receive placebo or 300 mg iP4 3 d postovulation (D3). Growth and vascularization of luteal tissue were evaluated by ultrasonography. Blood samples were collected from 3 d postovulation to 3 d after luteolysis, and P4 plasma concentrations were measured by radioimmunoassay. On days 3, 5, 7, 9, 11, 13, and 16 luminal endometrial samples were taken using a cytologic brush. Transcript abundance was measured by qPCR. Structural luteolysis occurred 3 d earlier in cows receiving iP4 compared to the control group. Analyzing only cows that received iP4, those that presented early luteolysis (ie, ≤ D16) showed a decrease in CL area and P4 concentration after D5, compared to the control group. Cows that presented early luteolysis showed a reduced abundance of transcripts on D5 for the ESR2 gene and a greater abundance of transcripts for OXTR and ESR1 on D16, compared to cows that did not present early luteolysis. The iP4-induced early luteolysis can be explained by two nonexclusive possibilities: the activation of uterine mechanisms that trigger early secretion of endometrial PGF pulses and the formation of a subfunctional CL that is prone to early regression.

Historical records of orchid bees (Apidae: Euglossini) in Belém Endemism Center: species list of 92 years sampling / Registros históricos de abelhas-das-orquídeas (Apidae: Euglossini) no Centro de Endemismo Belém: lista de espécies de 92 anos de amostragens

Abstract The distribution of most species occurs in delimited regions with unique characteristics called "centers of endemism". In Eastern Amazon is located the Belém Endemism Center (BEC), one of the most intensely deforested in Brazilian Amazon. Here, we show information about orchid bee assemblages based on historical records from entomological collections. For each species, we calculated occurrence frequency and dominance, and we classified them in 3 statuses: common, intermediate or rare species. Curves of observed and estimated richness were built, based on Jackknife estimator. We found 1,257 specimens from 56 species, constituting records from 1917 to 2009, and one species is a new record for BEC. Higher number of specimens and species was concentrated in a few locations and surveys increased from the 70's. The results suggest a high richness of orchid bees in the BEC, although this scenario is far from what is expected for the entire area. The high occurrence of rare species may be related to their low representativeness in the collections, and the proximity between the areas had favored samplings. Even so, the species list and the conservation status presented here may be useful information in studies comparing past and current orchid bee fauna, and, allied to data on bees' responses to land use changes occurred in BEC over the years, can fit as a basis for defining priority areas for conservation.

Resumo A distribuição da maioria das espécies ocorre em regiões delimitadas com características únicas conhecidas como "centros de endemismo". Na Amazônia Oriental está localizado o Centro de Endemismo Belém (CEB), uma das áreas mais intensivamente desmatadas na Amazônia Brasileira. Aqui, apresentamos informações sobre assembleias de abelhas orquídeas baseadas em registros históricos de coleções entomológicas. Para cada espécie, foram calculadas a frequência de ocorrência e a dominância, classificando-as em 3 estados: espécies comuns, intermediárias ou raras. Foram geradas curvas de riqueza observada e estimada, com base no estimador Jackknife. Encontramos 1.257 espécimes de 56 espécies, constituindo registros de 1917 a 2009, e uma espécie é um novo registro no CEB. Maior número de espécimes e espécies foi concentrado em poucos locais, com intensificação nas coletas a partir dos anos70. Os resultados sugerem alta riqueza de abelhas orquídeas no CEB, embora esse cenário esteja longe do que é esperado para toda a área. A elevada ocorrência de espécies raras pode estar relacionada à baixa representatividade nas coleções e a proximidade entre as áreas favoreceu as amostragens. Mesmo assim, a lista de espécies e o estado de conservação aqui apresentados podem ser informações úteis em estudos interessados em comparar a fauna passada e atual de abelhas das orquídeas e, aliado a dados sobre as respostas das abelhas às mudanças de uso da terra ocorridas no CEB ao longo dos anos, podem servir de base para definição de áreas prioritárias para conservação.

Cytobrush: A tool for sequential evaluation of gene expression in bovine endometrium.

Aims were to (i) compare specific transcript abundance between endometrial samples collected by transcervical biopsy and cytobrush and (ii) measure the abundance of endometrial transcripts involved in PGF2α synthesis in samples collected by cytobrush. In Experiment 1, endometrial samples were taken transcervically by cytobrush and biopsy 10 days after ovulation. Compared to biopsy samples, abundance of transcripts for MSTN, AKR1C4 and PGR was similar, VIM, FLT1 and PTGES was lower (p < .05) and KRT18 and CD3D was greater in cytobrush samples (p < .05). Thus, there was an enrichment of epithelial and immune cells in the cytobrush samples. In Experiment 2, endometrial samples were collected by cytobrush on days 10, 13, 16 and 19 after ovulation. Abundance of PGR2 mRNA was maximum on day 10 then decreased (p < .05). Abundance of ESR1 decreased gradually from day 10 to day 16 then increased again on day 19. The greatest abundance of OXTR was noted on day 19. The sequential alterations in abundance of these transcripts are consistent with the release of PGF2α associated with luteolysis. In summary, cytobrush sampling provides representative, physiologically relevant samples of the luminal epithelium in cattle.

Cerebral hemodynamics with intra-aortic balloon pump: business as usual?

OBJECTIVE: Intra-aortic balloon pump (IABP) is commonly used as mechanical support after cardiac surgery or cardiac shock. Although its benefits for cardiac function have been well documented, its effects on cerebral circulation are still controversial. We hypothesized that transfer function analysis (TFA) and continuous estimates of dynamic cerebral autoregulation (CA) provide consistent results in the assessment of cerebral autoregulation in patients with IABP. APPROACH: Continuous recordings of blood pressure (BP, intra-arterial line), end-tidal CO2, heart rate and cerebral blood flow velocity (CBFV, transcranial Doppler) were obtained (i) 5 min with IABP ratio 1:3, (ii) 5 min, starting 1 min with the IABP-ON, and continuing for another 4 min without pump assistance (IABP-OFF). Autoregulation index (ARI) was estimated from the CBFV response to a step change in BP derived by TFA and as a function of time using an autoregressive moving-average model during removal of the device (ARI t ). Critical closing pressure and resistance area-product were also obtained. MAIN RESULTS: ARI with IABP-ON (4.3 ± 1.2) were not different from corresponding values at IABP-OFF (4.7 ± 1.4, p = 0.42). Removal of the balloon had no effect on ARI t , CBFV, BP, cerebral critical closing pressure or resistance area-product. SIGNIFICANCE: IABP does not disturb cerebral hemodynamics. TFA and continuous estimates of dynamic CA can be used to assess cerebral hemodynamics in patients with IABP. These findings have important implications for the design of studies of critically ill patients requiring the use of different invasive support devices.

Visceral Marek's disease in white-peafowl (Pavo cristatus) / Doença de Marek visceral em pavão-branco (Pavo cristatus)

Marek's disease (MD) is a lymphoproliferative disorder caused by Gallid herpesvirus 2 (MDV) that infects mainly domestic gallinaceous birds although wild birds may occasionally be affected. The current report describes the anatomopathological and molecular findings of a case of MD in a white-peafowl (Pavo cristatus). The signs included apathy, hyporexia, and diarrhea. Grossly, 0.5 to 1.5cm in diameter, yellow, soft nodules were observed in the skeletal muscle, lung, kidney, air sacs, small intestine, heart, ovary, ventriculus, and proventriculus. Microscopically, numerous atypical round neoplastic cells were noted. The molecular detection of MDV DNA was implemented to amplify part of the meq gene and products were sequenced for the phylogenetic analysis. Template DNA was obtained from tissues of the affected bird and from blood of all the gallinaceous birds of the Zoo. The expected amplicon for the partial amplification of MDV meq gene was obtained and the amplicons were sequenced. Sequences obtained enabled grouping the strain (accession no. KT768121) with MDV serotype 1 strains from the GenBank. Based on the anatomopathological and molecular findings, the diagnosis of MD in a white-peafowl was reached, and to the authors' knowledge, no previous report regarding MD was published in Pavo cristatus.(AU)

Doença de Marek (MD) é uma desordem linfoproliferativa causada pelo Gallid herpesvirus 2 (MDV), que infecta principalmente galináceos domésticos, porém aves silvestres podem ser ocasionalmente afetadas. O presente relato descreve os achados anatomopatológicos e moleculares de um caso de MD em um pavão-branco (Pavo cristatus). Os sinais clínicos incluíram apatia, hiporexia e diarreia. Macroscopicamente, foram observados nódulos macios, de 0,5 a 1,5cm de diâmetro, no músculo esquelético, no pulmão, nos rins, nos sacos aéreos, no intestino delgado, no coração, no ovário, no ventrículo e no proventrículo. Microscopicamente, numerosas células redondas neoplásicas atípicas foram notadas. A detecção molecular do DNA do MDV foi implementada para amplificar parte do gene meq, e os produtos foram sequenciados para análise filogenética. DNA foi obtido de tecidos de aves afetadas e do sangue de todos os galináceos do zoológico. A esperada amplificação de parte do gene meq de MDV amplificado foi ampliada e sequenciada. As sequências obtidas permitiram o agrupamento da cepa (acesso KT768121) com cepas do sorotipo 1 de MDV do GenBank.. O diagnóstico de MD em pavão-branco foi obtido com base nos achados anatomopatológicos e moleculares e, pelo conhecimento dos autores, não há relatos anteriores publicados de MD em Pavo cristatus.(AU)

The Receptive Endometrial Transcriptomic Signature Indicates an Earlier Shift from Proliferation to Metabolism at Early Diestrus in the Cow.

This study aimed to characterize the endometrial transcriptome and functional pathways overrepresented in the endometrium of cows treated to ovulate larger (≥13 mm) versus smaller (≤12 mm) follicles. Nelore cows were presynchronized prior to receiving cloprostenol (large follicle [LF] group) or not (small follicle [SF] group), along with a progesterone (P4) device on Day (D) -10. Devices were withdrawn and cloprostenol administered 42-60 h (LF) or 30-36 h (SF) before GnRH agonist treatment (D0). Tissues were collected on D4 (experiment [Exp.] 1; n = 24) or D7 (Exp. 2; n = 60). Endometrial transcriptome was obtained by RNA-Seq, whereas proliferation and apoptosis were assessed by immunohistochemistry. Overall, LF cows developed larger follicles and corpora lutea, and produced greater amounts of estradiol (D-1, Exp. 1, SF: 0.7 ± 0.2; LF: 2.4 ± 0.2 pg/ml; D-1, Exp. 2, SF: 0.5 ± 0.1; LF: 2.3 ± 0.6 pg/ml) and P4 (D4, Exp. 1, SF: 0.8 ± 0.1; LF: 1.4 ± 0.2 ng/ml; D7, Exp. 2, SF: 2.5 ± 0.4; LF: 3.7 ± 0.4 ng/ml). Functional enrichment indicated that biosynthetic and metabolic processes were enriched in LF endometrium, whereas SF endometrium transcriptome was biased toward cell proliferation. Data also suggested reorganization of the extracellular matrix toward a proliferation-permissive phenotype in SF endometrium. LF endometrium showed an earlier onset of proliferative activity, whereas SF endometrium expressed a delayed increase in glandular epithelium proliferation. In conclusion, the periovulatory endocrine milieu regulates bovine endometrial transcriptome and seems to determine the transition from a proliferation-permissive to a biosynthetic and metabolically active endometrial phenotype, which may be associated with the preparation of an optimally receptive uterine environment.

Uso da técnica de swim-up para a remoção do Vírus da Artrite Encefalite Caprina do sêmen de reprodutores infectados / Use of swim-up technique for removal of the Caprine Arthritis Encephalitis Virus in the semen of infect bucks

Neste estudo, 67 ejaculados foram avaliados, antes e depois da técnica de swim-up, em relação à qualidade seminal e à presença do CAEV. Das 67 amostras testadas por PCRn, antes do swim-up, 47 (70,15%) foram positivas para o DNA pró-viral. No entanto, quatro amostras adicionais foram positivas ao RT-nested PCR após o swim-up, o que permite dizer que, pelo menos, 76,12% (51/67) delas estavam infectadas antes da lavagem. Todavia, em 23,88% (16/67) das amostras não foi detectada a presença do CAEV. Após a aplicação da técnica de swim-up, constatou-se, pela PCRn e RT-nested PCR, que houve uma redução significativa (χ²= 9,078; p<0,001) da presença do CAEV nas amostras seminais, pois 28 de 51 amostras positivas resultaram livres do vírus (54,90%), tanto para DNA pró-viral quanto para o vírus livre. Em relação à motilidade individual progressiva (MIP) e vigor espermático obtidos antes e depois da técnica de swim-up, observou-se uma diminuição significativa em suas médias, sendo o MIP de 86,42% para 71,49%, já o vigor espermático de 4,16 para 3,93. Conclui-se que a eliminação do CAEV no sêmen é de caráter intermitente, e que a associação da PCRn e RT-nested PCR é uma opção segura para a certificação sanitária individual das amostras seminais quanto à presença ou ausência do CAEV. Finalmente, a técnica de swim-up promove uma redução na infectividade de amostras de sêmen contaminadas, e, além disso, é possível promover a recuperação de espermatozoides viáveis.

In this study, 67 ejaculates were assessed before and after the swim-up technique in relation to semen quality and presence of CAEV. Of the 67 samples tested by Nested PCR, before swim-up 47 (70.15%) were positive for viral DNA. Furthermore, four additional samples were positive for RT-nested PCR after swim-up, which allows us to affirm that at least 76.12% (51/67) were infected before washing. However, 23.88% (16/67) of the samples did not detect the presence of CAEV. After application of the swim-up technique it was found, by Nested PCR and RT-nested PCR, that there was a significant decrease (χ² = 9.078, p <0.001) in the presence of CAEV in semen samples, once 28 of 51 positive samples were free from the virus (54.90%) for both proviral DNA and the free form of the virus. Regarding individual progressive motility (IPM) and spermatic vigor obtained before and after the swim-up technique, a significant decrease was observed in the average, being 86.42% of the IPM to 71.49% and the spermatic vigor from 4.16 for the 3.93. It is concluded that the removal of CAEV in semen has an intermittent character, and the combination of PCR and RT-nested PCR is a safe option for health certification of individual semen samples for the presence or absence of CAEV. Finally, the swim-up technique promotes a reduction in the infectivity of contaminated semen samples, and it is possible to promote the recovery of high individual progressive motility sperm and sperm vigor.

Uso da técnica de swim-up para a remoção do Vírus da Artrite Encefalite Caprina do sêmen de reprodutores infectados / Use of swim-up technique for removal of the Caprine Arthritis Encephalitis Virus in the semen of infect bucks

Neste estudo, 67 ejaculados foram avaliados, antes e depois da técnica de swim-up, em relação à qualidade seminal e à presença do CAEV. Das 67 amostras testadas por PCRn, antes do swim-up, 47 (70,15%) foram positivas para o DNA pró-viral. No entanto, quatro amostras adicionais foram positivas ao RT-nested PCR após o swim-up, o que permite dizer que, pelo menos, 76,12% (51/67) delas estavam infectadas antes da lavagem. Todavia, em 23,88% (16/67) das amostras não foi detectada a presença do CAEV. Após a aplicação da técnica de swim-up, constatou-se, pela PCRn e RT-nested PCR, que houve uma redução significativa (²= 9,078, p<0,001) da presença do CAEV nas amostras seminais, pois 28 de 51 amostras positivas resultaram livres do vírus (54,90%), tanto para DNA pró-viral quanto para o vírus livre. Em relação à motilidade individual progressiva (MIP) e vigor espermático obtidos antes e depois da técnica de swim-up, observou-se uma diminuição significativa em suas médias, sendo o MIP de 86,42% para 71,49%, já o vigor espermático de 4,16 para 3,93. Conclui-se que a eliminação do CAEV no sêmen é de caráter intermitente, e que a associação da PCRn e RT-nested PCR é uma opção segura para a certificação sanitária individual das amostras seminais quanto à presença ou ausência do CAEV. Finalmente, a técnica de swim-up promove uma redução na infectividade de amostras de sêmen contaminadas, e, além disso, é possível promover a recuperação de espermatozoides viáveis.

In this study, 67 ejaculates were assessed before and after the swim-up technique in relation to semen quality and presence of CAEV. Of the 67 samples tested by Nested PCR, before swim-up 47 (70.15%) were positive for viral DNA. Furthermore, four additional samples were positive for RT-nested PCR after swim-up, which allows us to affirm that at least 76.12% (51/67) were infected before washing. However, 23.88% (16/67) of the samples did not detect the presence of CAEV. After application of the swim-up technique it was found, by Nested PCR and RT-nested PCR, that there was a significant decrease (² = 9.078, p <0.001) in the presence of CAEV in semen samples, once 28 of 51 positive samples were free from the virus (54.90%) for both proviral DNA and the free form of the virus. Regarding individual progressive motility (IPM) and spermatic vigor obtained before and after the swim-up technique, a significant decrease was observed in the average, being 86.42% of the IPM to 71.49% and the spermatic vigor from 4.16 for the 3.93. It is concluded that the removal of CAEV in semen has an intermittent character, and the combination of PCR and RT-nested PCR is a safe option for health certification of individual semen samples for the presence or absence of CAEV. Finally, the swim-up technique promotes a reduction in the infectivity of contaminated semen samples, and it is possible to promote the recovery of high individual progressive motility sperm and sperm vigor.

AVE 0991, a non-peptide mimic of angiotensin-(1-7) effects, attenuates pulmonary remodelling in a model of chronic asthma.

BACKGROUND AND PURPOSE: AVE 0991 (AVE) is a non-peptide compound, mimic of the angiotensin (Ang)-(1-7) actions in many tissues and pathophysiological states. Here, we have investigated the effect of AVE on pulmonary remodelling in a murine model of ovalbumin (OVA)-induced chronic allergic lung inflammation. EXPERIMENTAL APPROACH: We used BALB/c mice (6-8 weeks old) and induced chronic allergic lung inflammation by OVA sensitization (20 µg·mouse(-1) , i.p., four times, 14 days apart) and OVA challenge (1%, nebulised during 30 min, three times per·week, for 4 weeks). Control and AVE groups were given saline i.p and challenged with saline. AVE treatment (1 mg·kg(-1) ·per day, s.c.) or saline (100 µL·kg(-1) ·per day, s.c.) was given during the challenge period. Mice were anaesthetized 72 h after the last challenge and blood and lungs collected. In some animals, primary bronchi were isolated to test contractile responses. Cytokines were evaluated in bronchoalveolar lavage (BAL) and lung homogenates. KEY RESULTS: Treatment with AVE of OVA sensitised and challenged mice attenuated the altered contractile response to carbachol in bronchial rings and reversed the increased airway wall and pulmonary vasculature thickness and right ventricular hypertrophy. Furthermore, AVE reduced IL-5 and increased IL-10 levels in the BAL, accompanied by decreased Ang II levels in lungs. CONCLUSIONS AND IMPLICATIONS: AVE treatment prevented pulmonary remodelling, inflammation and right ventricular hypertrophy in OVA mice, suggesting that Ang-(1-7) receptor agonists are a new possibility for the treatment of pulmonary remodelling induced by chronic asthma.

Grupo Cooperativo Iberoamericano de Medicina Transfusional. Programa Consulta al Experto. El uso adecuado de la sangre y sus alternativas en cirugía / Ibero-American Cooperative Group on Transfusion Medicine. Consulting the Expert. Appropiate use of blood products and its alternatives in surgical procedures

La transfusión de productos sanguíneos tiene como objetivo restaurar o mantener la capacidad de transportar oxígeno, y la homeostasis del volumen sanguíneo. En los últimos años se han introducido mejoras en las indicaciones de los productos sanguíneos ya que se ha demostrado que su uso supone un riesgo para el paciente y también es costoso para el sistema de salud. Hay grupos que están trabajando con la práctica de la "cirugía sin sangre". El propósito principal de realizar intervenciones utilizando poca sangre ("cirugía sin sangre") está enfocada en minimizar las pérdidas y reducir o eliminar la exposición a los componentes sanguíneos alogeneicos. Los riesgos asociados con las transfusiones de sangre han sido bien documentados, y el objetivo de los centros de "cirugía sin sangre" es evitar complicaciones y el uso innecesario de las transfusiones. Hay una estrategia universal de uso conservador de la sangre que es aplicable a todos los pacientes y poblaciones. Factores tales como las condiciones preexistentes van a cambiar el enfoque en casos específicos. Un protocolo de cirugía exitosa con poca pérdida de sangre requiere trabajo en equipo y la cooperación entre el banco de sangre, hematólogos, cirujanos y anestesiólogos, para asegurar que se cumplan los objetivos de minimizar la pérdida de sangre y evitar las transfusiones, sin poner en riesgo la vida y el bienestar del paciente.

Immunization of mice with Lactobacillus casei expressing a beta-intimin fragment reduces intestinal colonization by Citrobacter rodentium.

Enteropathogenic Escherichia coli (EPEC) is a common cause of diarrhea in children from developing countries. Intimate adhesion of the bacteria to intestinal cells occurs via binding of the adhesin intimin to the TIR receptor exposed on cell surfaces. Here, Lactobacillus casei expressing a fragment of ß-intimin (L. casei-Int(cv)) was tested as mucosal vaccines in mice against intestinal colonization with the murine pathogen Citrobacter rodentium. Oral or sublingual immunization of C57BL/6 mice with L. casei-Int(cv) induced anti-Int(cv) IgA in feces but no IgG in sera. Conversely, anti-Int(cv) IgG was induced in the sera of mice after sublingual immunization with purified Int(cv). All vaccines were able to decrease C. rodentium recovery from feces. However, this reduction was more evident and sustained over time in mice immunized with L. casei-Int(cv) by the sublingual route. These mice also displayed an increase in interleukin 6 (IL-6) and gamma interferon (IFN-γ) secretion by spleen cells 10 days after infection. Additionally, oral or sublingual immunization of C3H/HePas mice, which are highly susceptible to C. rodentium infection, with L. casei-Int(cv) induced anti-Int(cv) antibodies and significantly increased survival after challenge. Immunohistological analysis of colon sections revealed that C. rodentium was located in deep fractions of the tissue from C3H/HePas mice immunized with L. casei whereas superficial staining was observed in colon sections from mice immunized with L. casei-Int(cv.) The results indicate that vaccines composed of L. casei expressing intimin may represent a promising approach and that the C3H/HePas infection model with C. rodentium can be used to evaluate potential vaccines against EPEC.

Accuracy of routine diagnostic tests used in paracoccidioidomycosis patients at a university hospital.

The identification of appropriate laboratory measures to confirm clinical hypotheses is important in routine paracoccidioidomycosis medical care. The clinical records and laboratory reports of 401 paracoccidioidomycosis patients attended at the Tropical Diseases Area, Faculdade de Medicina de Botucatu, from 1974 to 2008 were reviewed. Direct mycological (DM), cell block (CB), histopathological (HP), and double immunodiffusion (DID) tests were evaluated before treatment. Typical Paracoccidioides brasiliensis yeast forms were observed in clinical specimens of 86% of the patients, but 14% were detected only by serological test. DM of 51 different tissue specimens produced 74.5% sensitivity, and 62.5% sensitivity was observed in 112 sputum samples. CB in 483 sputum samples generated 55.3% sensitivity. HP performed in 239 samples from different tissues revealed 96.7% sensitivity. Serology carried out in 351 patients and 200 healthy controls provided 90.0% sensitivity, 100.0% specificity, 100.0% positive predictive value, 85.1% negative predictive value and 93.6% accuracy. Comparisons of laboratory measurements performed in the same patient showed that sensitivity decreases from HP to DID to CB and DM, with the last two assays providing similar sensitivities. This study demonstrated that P. brasiliensis identification by HP, CB, and/or DM associated with DID is sufficient to establish the laboratorial diagnosis of paracoccidioidomycosis in practically all cases.

Trends in adjuvant development for vaccines: DAMPs and PAMPs as potential new adjuvants

Aluminum salts have been widely used in vaccine formulations and, after their introduction more than 80 years ago, only few vaccine formulations using new adjuvants were developed in the last two decades. Recent advances in the understanding of how innate mechanisms influence the adaptive immunity opened up the possibility for the development of new adjuvants in a more rational design. The purpose of this review is to discuss the recent advances in this field regarding the attempts to determine the molecular basis and the general mechanisms underlying the development of new adjuvants, with particular emphasis on the activation of receptors of innate immune recognition. One can anticipate that the use of these novel adjuvants will also provide a window of opportunities for the development of new vaccines.

Estudo químico e da atividade biológica cardiovascular do óleo essencial de folhas de Alpinia zerumbet (Pers.) B.L.Burtt & R.M.Sm. em ratos / Phytochemistry and cardiovascular biological activity of the essential oil from leaves of Alpinia zerumbet (Pers.) B.L. Burtt & R.M.Sm. in rats

A espécie vegetal Alpinia zerumbet (Pers.) B.L.Burtt & R.M. Sm. é popularmente empregada para o tratamento de diversas enfermidades, entre elas a hipertensão. Avaliar a composição química, a atividade antihipertensiva e ação na hipertrofia cardíaca do óleo essencial das folhas de Alpinia zerumbet (OEAZ) em ratos foram os objetivos deste estudo. O OEAZ, obtido por hidrodestilação em aparelho Clevenger, teve sua composição química analisada em cromatografia gasosa acoplada à espectrometria de massas (CG-EM). Foram identificados 14 constituintes, sendo terpinen-4-ol (37,45 por cento) o majoritário, seguido pelos óxido de cariofileno (7,56 por cento), trans-hidrato de sabineno (6,61 por cento) e 1,8-cineol (4,02 por cento). A avaliação cardiovascular foi feita após o tratamento crônico de ratos espontaneamente hipertensos (SHR) e seus respectivos controles, ratos Wistar-Kyoto (WKY). Os dados hemodinâmicos revelaram redução da pressão arterial média (PAM) no grupo tratado (SHRP: 160 ± 7 mm Hg; p<0,01) em relação ao não tratado (SHR: 180 ± 5 mm Hg). A relação entre peso do ventrículo esquerdo e peso corporal (VE/PC) do SHRP (2,50 ± 0,03 mg g-1; p<0,01) mostrou-se inferior ao SHR (2,61 ± 0,01 mg g-1), confirmando a redução da hipertrofia cardíaca (HC). Os dados de PAM e VE/PC dos animais SHRP foram estatisticamente diferentes quando comparados com os ratos controle (WKY: 116 ± 2 mm Hg e WKYP: 119 ± 4 mm Hg; p<0,05; WKY: 2,15 ± 0,04 mg g-1 e WKYP: 2,17 ± 0,04 mg g-1 ; p<0,01), indicando não ter havido normalização dos mesmos. Conclui-se que o tratamento crônico com OEAZ foi capaz de determinar redução, mas não a normalização, da PAM e da HC de ratos SHR, provavelmente pela presença dos componentes terpinen-4-ol e 1,8-cineol. Estudos com doses maiores ou período de tratamento superior são necessários para avaliar a possibilidade de o OEAZ normalizar os parâmetros analisados (PAM e HC).

Alpinia zerumbet (Pers.) B.L. Burtt & R.M.Sm. is traditionally employed to treat several diseases such as hypertension. The aim of this study was to evaluate the chemical composition, the anti-hypertensive activity and the capacity to reduce cardiac hypertrophy of the essential oil of A. zerumbet leaves (EOAZ) in rats. EOAZ was obtained through hydrodistillation in Clevenger apparatus and its chemical composition was analyzed by gas chromatography-mass spectrometry (GC-MS). Several constituents (14) were identified, terpen-4-ol (37.45 percent) being the major component, followed by caryophyllene oxide (7.56 percent), trans-sabinene hydrate (6.61 percent) and 1,8-cineol (4.02 percent). The cardiovascular effect was investigated after chronic treatment with spontaneously hypertensive rats (SHR) and their respective controls, Wistar-Kyoto rats (WKY). The treated group showed a lower mean arterial pressure (MAP) (SHRP: 160 ± 7 mm Hg; p<0.01) than the untreated group (SHR: 180 ± 5 mm Hg). The ratio of left ventricle-to-body weight (LV/BW) for SHRP was lower (2.504 ± 0.03 mg g-1; p<0.01) than that for SHR (2.162 ± 0.01 mg g-1), confirming the cardiac hypertrophy (CH) reduction. There were significant differences in MAP and CH between SHRP animals and control rats (WKY: 116 ± 2 mm Hg and WKYP: 119 ± 4 mm Hg; p<0.05. WKY: 2.152 ± 0.04 mg g-1 and WKYP: 2.168 ± 0.04 mg g-1; p<0.01), indicating that these values were not normalized. Those data showed that the chronic treatment with EOAZ reduces MAP and CH in SHR probably due to the presence of the compounds terpinen-4-ol and 1,8-cineol. Studies with higher doses or longer treatment periods are necessary to evaluate whether EOAZ can reduce the analyzed parameters (MAP and CH) to normal values.