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This paper presents a surface and subsurface geochemical survey of the Buracica EOR-CO2 field onshore Brazil. We adopted a methodology coupling the stable isotopes of carbon with noble gases to investigate the adequacy of geochemical monitoring to track deep fluid leakage at the surface. Three campaigns of CO2 flux and concentration in soils were performed to understand the CO2 variability across the field. The distribution of the CO2 soil contents between 0.8 and 14% is in great part controlled by the properties of the soil, with a first-order topographic dependency. These results, together with a δ13CCO2 between –15 and –23‰, suggest that the bulk of the soil CO2 flux is biological. The gas injected and produced at numerous wells across the field showed a great spatial and somewhat temporal heterogeneity with respect to molecular, δ13CCO2 and noble gas compositions. This heterogeneity is a consequence of the EOR-induced sweeping of the petroleum fluids by the injected CO2, producing a heterogeneous mixing controlled by the production scheme and the distribution in reservoir permeability. In light of the δ13CCO2 found in the reservoir, the stable isotopic composition of carbon was insufficient to track CO2 leaks at the surface. We demonstrate how noble gases may be powerful leak discriminators, even for CO2 abundances in soils in the bottom range of the biological baseline (~1%). The results presented in this study show the potential of geochemical monitoring techniques, involving stable isotopes and noble gases at the reservoir and soil levels, for tracing CO2 in CCS projects. Le monitoring géochimique du gisement de Buracica, qui produit des hydrocarbures par récupération assistée et injection de dioxyde de carbone, est présenté dans cet article. Une méthodologie permettant de coupler l’utilisation des isotopes stables du carbone et des isotopes des gaz rares pour étudier la faisabilité de traçage d’une fuite de CO2 du réservoir à la surface est détaillée. Les campagnes de prélèvement se sont déroulées sur trois périodes en deux ans, permettant de mesurer la variabilité des teneurs en CO2 dans les sols et dans les réservoirs pétroliers. La distribution du CO2 en surface varie de 0,8 à 14 % et peut être contrôlée en partie par les propriétés des sols et la topographie. Les résultats isotopiques du δ13CCO2 varient entre –15 et –23 ‰ et suggèrent que le flux de CO2 soit principalement biogénique. Le gaz injecté, puis produit à Buracica, est hétérogène en composition de par la récupération tertiaire du gisement induisant une infiltration de CO2 à des niveaux variables et les variations de perméabilité au sein même du réservoir. Les résultats montrent qu’il est, dans ce cas, difficile de tracer le CO2 du réservoir à la surface en fonction des valeurs isotopiques δ13CCO2. Notre approche démontre le fort potentiel des gaz rares pour discriminer des mélanges, même à des teneurs faibles de CO2 (~1 %). Les résultats illustrent le potentiel de ces nouvelles techniques de monitoring géochimiques pour suivre le CO2 dans les projets de séquestration combinant les signatures du carbone stable avec celles des gaz rares.

Background: In prolonged laparoscopic procedures, hypothermia is frequently observed. The possible influence of the vasodilating action of CO(2), due to its increased levels in the blood during the laparoscopic procedures, has yet to be studied. The objective of this study was, therefore, to evaluate body temperature patterns in pigs subjected to pneumoperitoneum with CO(2).Methods: Thirty male pigs were allocated into three groups of ten animals each: group I, anesthetic procedure and abdominal puncture only; group II, the same as for group I and insufflation with CO(2); and group III, the same as for group I and insufflation with medical grade compressed air. After anesthetic induction and surgical preparation, rectal and esophageal temperatures were measured every 10 min. Blood was collected during the experiment for the gasometric measurement of pCO(2). Animals were insufflated with no gas loss and were kept anesthetized for 180 min. For statistical analysis, Friedman and Kruskal-Wallis tests were used at a level of significance of 95% (P < 0.05).Results: Animals in groups I and II (P = 0.000) had a statistically significant drop in both esophageal and rectal temperatures during the experiment, but not animals in group III. However, when the groups were compared among themselves, no statistically significant differences were found at any of the times measured. A statistically significant drop in pCO(2) levels was observed for groups I and III, but not for animals in groups II.Conclusions: The use of CO(2) did not significantly affect body temperature variation in pigs subjected to pneumoperitoneum. However, CO(2) produced a temperature drop pattern different than that of compressed air, indicating that CO(2) may lead to thermoregulatory changes and influence the peripheral temperature drop. [ABSTRACT FROM AUTHOR]

Abstract: The political and social acceptability of the geological storage of CO2 depends on the existence of reliable site surveillance technologies. The ground surface is the widest available area over which a permanent control of possible leaks may be performed, and also a privileged area where to investigate to which extent a leak at the reservoir depth may disturb the superficial environment. In this article, we suggest several surface surveillance guidelines inspired from data originating from an EOR- CO2 oil field case study where an extensive survey of surface and reservoir gas has been performed. This study shows that the gases that may leak from a CO2 geological storage can have a wide range of compositions, both molecular and isotopic. The complexity in the gas source is further impacted by the likely changes in gas (or more generally fluid) composition on the leaking pathway. With such heterogeneity and variability in mind, the task of monitoring a leak at the surface becomes a serious challenge. Furthermore, comparing natural background fluxes of CO2 with expected fluxes in worst case leak scenarios shows that both have comparable amplitudes (∼0.4 mol/m2/day). The flux of CO2 itself, or the CO2 content of the soil are of limited interest with respect to a leak detection. Using calculations based on the Dusty Gas Model, we show that the coupling of carbon isotopes and the noble gases with flux measurements is a powerful monitoring method, able to discriminate the flux contribution of a leak in the flux range of the natural baseline. [Copyright &y& Elsevier]

An attempt was made to quantify the influence of prey scarcity on longevity and on the reproductive variables of females of the predator Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae). The predators received prey ( Alabama argillacea) of different sizes, viz., 50 mg (small prey) or 220 mg (large prey), for a period of 24 h, daily or every 2, 4, 8 or 16 days. In an additional trial, the predators were provided only with water or kept without food, in which latter condition they lived a significantly shorter time. Individuals that received large prey in excess (every day) and at 16-day intervals lived significantly less, while the longevity of those that fed on small prey did not differ significantly between feeding intervals. Predators that fed less frequently laid fewer eggs, with increased intervals between clutches. The results were more marked in females fed on small prey; however, evidence of trade-off between reproduction and survival was detected only in predators that received large prey. The results suggest a type of trade-off, where longevity is maintained at the expense of reproduction. This strategy can contribute to the establishment and survival of P. nigrispinus in an environment of fluctuating food supply. [ABSTRACT FROM AUTHOR]

The food extraction by adult females of Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae) from 3 rd instar Alabama argillacea (Hübner) larvae was evaluated in the laboratory. The densities studied were converted to weight of prey offered: 15.7 (one larva); 44.1 (three larvae); 71.4 (five larvae); 117.4 (seven larvae) and 148.8 mg (nine larvae). The adult weight, the amount of food consumed, the weight of remaining larvae and the time taken to ingest the food were recorded every 24 h during 9 days to estimate the percentage of food consumed, the quantity and the percentage of food extracted from each larva, the quantity of food extracted per minute and the relative consumption rate. The quantity of food consumed (88.5% to 35.5%) and extracted (90.8% to 35.5%) by P. nigrispinus females from each larva decreased as the food available increased. The time spent by P. nigrispinus to ingest food and the quantity of remaining food were similar during the 9-day period of the adult stage of this predator. No interactions were observed between the age of females of this predator and the prey density. The weight gained (1.2–5.8 mg/9 days), body weight (35.5–42.3 mg) and the relative consumption rate (0.4–1.4 mg/mg/day) by females of this predator increased at a declining rate with the quantity of food available. P. nigrispinus may change its predatory behavior as a function of food available. [ABSTRACT FROM AUTHOR]

During the past decade, the Portuguese political system has been under close scrutiny. As in other Western democracies, the topic of citizens' dissatisfaction with democratic institutions has been at the centre of a wide debate amongst public opinion and politicians. This debate has focused on parliament and parliamentary reform has often been seen as a way of enhancing parliament's image. However, as this note will show, there is no clear evidence that the Portuguese are dissatisfied with this institution. Indeed, this is still an under-researched area. Why is there an assumption that the Portuguese parliament has a negative image? And why should this have consequences for parliamentary reform? In the context of the creation of a Portuguese Election Study, this note aims to raise questions for future research on this topic. The note will look into the concept of legislative support, assess the evidence available on the Portuguese parliament's popularity and suggest indicators that would help to clarify the Portuguese views on this. It will also explore the connection between legislative support and the role of parliament, to enquire whether popularity indicators can make a case for parliamentary reform. [ABSTRACT FROM PUBLISHER]

PURPOSE: To evaluate, in dogs, the biliary sphincter subjected to dilation by hydrostatic balloon by the point of view of structural alterations of the papilla and the biochemestry and bacterial contamination of the bile. METHODS: Twenty dogs were submitted to laparotomy, duodenotomy, and enlargement of the major duodenal papilla- GA(n=10) - with balloon of 8mm inflated with pressure of 0,5atm, during 2 minutes or to the sham procedure - GB(n=10). Blood samples collected on times t(0day), t(7days) and t(28days) were subjected to dosages of alkaline phosphatase (ALP) and gamma-glutamyltransferase (GGT) for cholestasis evaluation. The collected material from the gall bladder at the same times were registered and numbered to be submitted to culture in BHI, blood agar (rich, non-selective element) and Mac Conkey (selective element for Gram-negative bacillus. On the 28th day three fragments of the papilla were tranversally cut by the choledoc axis 3mm from the duodenal papilla and the cuts, stained with hematoxylin-eosin and Masson's tricome, were evaluated according to their inflammatory reaction. RESULTS: The GGT and ALP averages on the three periods in the groups A and B did not show significant differences, not being characterizes the cholestasis. The bacterian contamination was significantly higher in GA (2,19) than in GB (1,96); the contamination was lower in the initial time compared with 7 and 28 days (t0

Várias substâncias identificáveis estão implicadas no processo de crescimento hepático, entre elas os fatores de crescimento. A maioria deles possui diferentes ações, estimulando a proliferação de células, ou mesmo inibindo na dependência de suas concentrações. Dentre os fatores de crescimento, ou somatomedinas, com ação sobre o fígado pode-se destacar: HGF; EGF; TGF-alpha; TGF b ; Interleucina 6; IGF; FGF; VEGF; KGF; HSS e ALR. A ação conjunta dos hormônios HGF, TGF-alpha, IL-6, TNF-alpha, norepinefrina, EGF, permite que insulina, glucagon e o próprio EGF manifestem seus efeitos. O HGF tem papel vital, talvez o principal "gatilho" deste processo, gerando um sinal endócrino que ativa fortemente a mitogênese nos hepatócitos já "preparados" pelo EGF, IL-6, insulina, matriz remanescente e outros, levando à síntese de DNA. Admite-se que o EGF também participe dos eventos iniciais do processo logo após a hepatectomia e que FGF, VEGF e KGF também participem dos eventos ligados à recomposição de outros tecidos.

O objetivo deste trabalho foi comparar, de modo prospectivo e casualizado, duas técnicas de sutura: uma com revestimento interno com dreno de PENROSE e outra utilizando a invaginação submucosa-mucosa nas anastomoses esôfago-esofágicas cervicais. Foram utilizados trinta e dois cães machos com pesos entre 8 e 10 Kg, distribuídos em dois grupos de dezesseis animais. Cada grupo foi dividido em dois subgrupos baseado nos períodos observados até o 7o e 14o dias de pós-operatórios. Em cada grupo, as anastomoses foram avaliadas em seus aspectos clínicos, macroscópicos e microscópicos. Nenhuma fístula ou óbito foi observado no pós-operatório. Todos os animais do grupo com revestimento interno com dreno de PENROSE apresentaram disfagia por tempo mínimo de sete dias, enquanto apenas quatro animais do grupo sutura com invaginação apresentaram disfagia transitória com duração máxima de dois dias. À microscopia óptica, foi observada deficiência na regeneração das túnicas submucosa e mucosa nas anastomoses com prótese. Os resultados observados permitem concluir que a anastomose esôfago-esofágica cervical com revestimento interno com dreno de PENROSE apresentou piores resultados do que a por invaginação.

O objetivo deste trabalho foi avaliar o efeito da heparinização regional durante a isquemia arterial temporária em coelhos. Os parâmetros avaliados foram a medida do tempo de tromboplastina parcial ativada (TTPa) e a presença ou ausência de trombos ou edema endotelial nos vasos estudados. Foram utilizados 40 coelhos Nova Zelândia, distribuídos em dois grupos de 20 animais. Após dissecção e reparo da artéria ilíaca esquerda, realizou uma arteriotomia e introduziu-se um cateter de polietileno para injeção de solução de heparina no grupo experimento ou de soro fisiológico no grupo controle. O TTPa foi medido no início do experimento e após 90 minutos de isquemia, não sendo encontrado diferenças significantes entre os valores iniciais e ao final do experimento. Na avaliação microscópica não foi encontrada a presença de trombos nos vasos dos animais de ambos os grupos. Em apenas um animal do grupo controle houve a ocorrência de espessamento intimal e desarranjo da camada média, interpretado como edema endotelial, porém este dado não foi significante. Concluiu-se que a injeção intra-arterial de heparina na dose de 60 UI/kg peso não provocou alterações significantes na coagulação sistêmica do coelho, e que na ausência de lesão endotelial não ocorreu trombose dos vasos estudados, com ou sem a utilização de heparina.

Public engagement has become a noticeable activity for parliaments across the world. However, we lack understanding of its role despite considerable developments in scholarly work on public engagement in the sciences and on deliberative and participatory democracy by social scientists. This article provides a framework to understand the significance of parliamentary public engagement and to evaluate its effectiveness. It explains how parliamentary public engagement has emerged because of a representational shift in who is doing the representing in parliament and in what is represented, following key societal changes. We define parliamentary public engagement, showing the importance of differentiating between the activity, its effects and broader democratic ideals. We identify information and education as the types of engagement activity most developed by parliaments, with much still to do in consultation and participation activities. The article finishes with a discussion of seven key challenges in developing and implementing effective institutional parliamentary public engagement practices., Competing Interests: No potential conflict of interest was reported by the authors., (© 2023 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

The importance of family farming in food systems worldwide is recognized by different international bodies, as well as the leading role played by women and the inequalities they face in this sector of activity. The most recent data from Portugal highlight the importance of this type of agriculture in this Southern European country. In 2019, 68% of the total agricultural workforce in the country was concentrated in family farming, with almost half of them being women. This high permanence of women in agriculture is the result of a long process of feminization on this sector that is similar to other contexts. Despite this strong feminization of family farming, there are few studies that portrait agricultural activity from the women's viewpoint, since the voice of men is always predominant in all references. Based on the exploratory qualitative data from two focus groups , carried out in two Portuguese inner regions, we intend to address the perceptions and meanings of a small group of women farmers regarding their activity, the role taken by them in agriculture and the difficulties they experience. Issues such as changes in agriculture and the sexual division of labor will also be addressed in this article. Within these groups, women work in agriculture is perceived as long, solitary and uncertain. Also, the public/private dichotomy is evident, with decision-making and public places dominated by men. A prevalence of the discourse of "masculinization" still exists with certain tasks being attributed to men (e.g., operations with machinery). Younger women (34 and 40 years old) tend to overcome these gender differences choosing agriculture as a profession and healthy and sustainable life for their families., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Gomes, Jesus, Rosa, Bandeira and Costa.)

Background: Morphology of the human cerebral cortex differs across psychiatric disorders, with neurobiology and developmental origins mostly undetermined. Deviations in the tangential growth of the cerebral cortex during pre/perinatal periods may be reflected in individual variations in cortical surface area later in life., Methods: Interregional profiles of group differences in surface area between cases and controls were generated using T1-weighted magnetic resonance imaging from 27,359 individuals including those with attention-deficit/hyperactivity disorder, autism spectrum disorder, bipolar disorder, major depressive disorder, schizophrenia, and high general psychopathology (through the Child Behavior Checklist). Similarity of interregional profiles of group differences in surface area and prenatal cell-specific gene expression was assessed., Results: Across the 11 cortical regions, group differences in cortical area for attention-deficit/hyperactivity disorder, schizophrenia, and Child Behavior Checklist were dominant in multimodal association cortices. The same interregional profiles were also associated with interregional profiles of (prenatal) gene expression specific to proliferative cells, namely radial glia and intermediate progenitor cells (greater expression, larger difference), as well as differentiated cells, namely excitatory neurons and endothelial and mural cells (greater expression, smaller difference). Finally, these cell types were implicated in known pre/perinatal risk factors for psychosis. Genes coexpressed with radial glia were enriched with genes implicated in congenital abnormalities, birth weight, hypoxia, and starvation. Genes coexpressed with endothelial and mural genes were enriched with genes associated with maternal hypertension and preterm birth., Conclusions: Our findings support a neurodevelopmental model of vulnerability to mental illness whereby prenatal risk factors acting through cell-specific processes lead to deviations from typical brain development during pregnancy., (Copyright © 2022 Society of Biological Psychiatry. Published by Elsevier Inc. All rights reserved.)

Zika virus (ZIKV) infection has caused severe unexpected clinical outcomes in neonates and adults during the recent outbreak in Latin America, particularly in Brazil. Congenital malformations associated with ZIKV have been frequently reported; nevertheless, the mechanism of vertical transmission and the involvement of placental cells remains unclear. In this study, we applied quantitative proteomics analysis in a floating explant model of chorionic villi of human placental tissues incubated with ZIKV and with ZIKV pre-adsorbed with anti-ZIKV envelope protein. Proteomic data are available via ProteomeXchange with identifier PXD025764. Altered levels of proteins were involved in cell proliferation, apoptosis, inflammatory processes, and the integrin-cytoskeleton complex. Antibody-opsonized ZIKV particles differentially modulated the pattern of protein expression in placental cells; this phenomenon may play a pivotal role in determining the course of infection and the role of mixed infections. The expression of specific proteins was also evaluated by immunoperoxidase assays. These data fill gaps in our understanding of early events after ZIKV placental exposure and help identify infection control targets., (Copyright © 2021 Elsevier B.V. All rights reserved.)

This study adapted and examined the psychometric properties of the Youth Life Orientation Test (YLOT)-a measure to evaluate dispositional optimism and pessimism-in a Brazilian sample of children and adolescents aged between 5 and 16 years. The factor structure, reliability, convergent validity, and invariance of the YLOT were evaluated within and across four age groups ( N 1 = 266, N 2 = 246, N 3 = 211, and N 4 = 253). Participants were 976 students (51.9% girls) who answered the YLOT and subjective well-being measures and 350 parents (51.6% female) who answered a parent-report measure of optimism. Confirmatory factor analyses suggested an oblique two-factor model best fit the data, indicating that optimism and pessimism are independent factors. These factors demonstrated adequate reliability and convergent validity evidence with well-being measures. Additionally, scalar invariance was obtained, and latent mean analyses indicated optimism decreases and pessimism increases in participants from 5 to 13 years old. These results support the use of YLOT in Brazil. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Experimental studies have demonstrated that methylphenidate (MPH) modulates the synaptic vesicle trafficking and synaptotagmin-1 (SytI) mRNA levels. SytI is a regulatory protein of the SNARE complex, a neurotransmitter exocytosis mediator. Despite this evidence, most SNARE complex-related genes have never been evaluated in attention-deficit/hyperactivity disorder (ADHD) pharmacogenetics. This study evaluates, for we believe the first time, polymorphisms on the SNARE complex-related genes STX1A (rs2228607), VAMP2 (26bp Ins/Del) and SYT1 (rs1880867 and rs2251214) on the response to immediate-release methylphenidate (IR-MPH) in a naturalistic sample of adults with ADHD. The sample comprised 433 subjects, of which 272 (62.8%) have completed the short-term IR-MPH treatment (at least 30 days). The main outcome measure was the categorical variable of short-term response to IR-MPH based on the Swanson, Nolan and Pelham Rating Scale version 4 (SNAP-IV), and on the clinical global impression-improvement scale. Additional analyses evaluated the percentage of SNAP-IV symptom reduction for each dimension as well as short- and long- (7 years) term treatment persistence. SYT1-rs2251214 was associated with the categorical short-term response to IR-MPH (P=0.006, P FDR =0.028), and with the percentage of inattention and oppositional defiant disorder symptoms reduction (P=0.007, P FDR =0.028 and P=0.017, P FDR =0.048, respectively). SYT1-rs2251214 was also associated with short-term treatment persistence (P=0.018, P FDR =0.048), and with months of treatment (P=0.002, P FDR =0.016) in the long-term protocol. Our findings suggest that SYT1-rs2251214 presents a broad influence in IR-MPH response variability in adults with ADHD, being involved with both symptom response and treatment persistence. If such findings are replicated, SytI could represent a key element in MPH pharmacodynamics in adults with ADHD.

Placental barrier regulates maternal-fetal interchange protecting the baby from damage caused by substances found in the uterine environment or circulating in the vascular system. Organophosphate (OP) pesticides are a paramount group of environmental pollutants used in intensive agriculture for protection against diseases and pests. While many studies have reported an increased risk of pregnancy alterations in pregnant women exposed to OPs, few have analyzed the effects caused by these pesticides in the placenta. Herein, we evaluated the effects of chlorpyrifos (CPF), one of the most widely used OP insecticides, on human placenta using in vitro and ex vivo exposure models. Villous cytotrophoblast cells isolated from normal human term placentas maintained their cell viability, differentiated into syncytiotrophoblast-like structures, and increased the expression of β-hCG, ABCG2, and P-gp in the presence of CPF at concentrations of 10 to 100μM. The same doses of CPF induced marked changes in chorionic villi samples. Indeed, CPF exposure increased stroma cell apoptosis, altered villi matrix composition, basement membrane thickness, and trophoblastic layer integrity. Histomorphological and ultrastructural alterations are compatible with those found in placentas where maternal-placenta injury is chronic and able to impair the placental barrier function and nutrient transport from mother to the fetus. Our study shows that placental ex vivo exposure to CPF produces tissue alterations and suggest that human placenta is a potential target of CPF toxicity. In addition, it highlights the importance of using different models to assess the effects of a toxic on human placenta., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The teratogenic mechanisms triggered by ZIKV are still obscure due to the lack of a suitable animal model. Here we present a mouse model of developmental disruption induced by ZIKV hematogenic infection. The model utilizes immunocompetent animals from wild-type FVB/NJ and C57BL/6J strains, providing a better analogy to the human condition than approaches involving immunodeficient, genetically modified animals, or direct ZIKV injection into the brain. When injected via the jugular vein into the blood of pregnant females harboring conceptuses from early gastrulation to organogenesis stages, akin to the human second and fifth week of pregnancy, ZIKV infects maternal tissues, placentas and embryos/fetuses. Early exposure to ZIKV at developmental day 5 (second week in humans) produced complex manifestations of anterior and posterior dysraphia and hydrocephalus, as well as severe malformations and delayed development in 10.5 days post-coitum (dpc) embryos. Exposure to the virus at 7.5-9.5 dpc induces intra-amniotic hemorrhage, widespread edema, and vascular rarefaction, often prominent in the cephalic region. At these stages, most affected embryos/fetuses displayed gross malformations and/or intrauterine growth restriction (IUGR), rather than isolated microcephaly. Disrupted conceptuses failed to achieve normal developmental landmarks and died in utero. Importantly, this is the only model so far to display dysraphia and hydrocephalus, the harbinger of microcephaly in humans, as well as arthrogryposis, a set of abnormal joint postures observed in the human setting. Late exposure to ZIKV at 12.5 dpc failed to produce noticeable malformations. We have thus characterized a developmental window of opportunity for ZIKV-induced teratogenesis encompassing early gastrulation, neurulation and early organogenesis stages. This should not, however, be interpreted as evidence for any safe developmental windows for ZIKV exposure. Late developmental abnormalities correlated with damage to the placenta, particularly to the labyrinthine layer, suggesting that circulatory changes are integral to the altered phenotypes.

The transcription factor Krüppel-Like Factor 6 (KLF6) has important roles in cell differentiation, angiogenesis, apoptosis, and proliferation. Furthermore, there is evidence that KLF6 is required for proper placental development. While oxygen is a critical mediator of trophoblast differentiation and function, the involvement of oxygen in the regulation of KLF6 expression remains unexplored. In the present study we examined the expression of KLF6 in placental tissue from uncomplicated and preeclamptic pregnancies, the latter often characterized by an inadequately perfused placenta. We also determined the effect of hypoxia and the involvement of Hypoxia-Inducible Factor 1α (HIF-1α) on the expression of KLF6 in cultured trophoblast cells and placental tissues. Results revealed that villous, interstitial and endovascular extravillous cytotrophoblasts from placentas from normal and preeclamptic pregnancies express KLF6. In addition, KLF6 immunoreactivity was higher in the placental bed of preeclamptic pregnancies than in those of uncomplicated pregnancies. We demonstrated that hypoxia induced an early and transient increase in KLF6 protein levels in HTR8/SVneo extravillous cytotrophoblast cells and in placental explants. Reoxygenation returned KLF6 protein to basal levels. Moreover, hypoxia-induced up-regulation of KLF6 expression was dependent on HIF-1α as revealed by siRNA knockdown in HTR8/SVneo cells. These results indicate that KLF6 may mediate some of the effects of hypoxia in placental development. The regulation of KLF6 protein levels by oxygen has significant implications for understanding its putative role in diseases affected by tissue hypoxia., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Denaturing high-performance liquid chromatography (DHPLC) has been described as a suitable method to study DNA polymorphisms. Here, cassava (Manihot esculenta Crantz) fermentation liquor was examined using DHPLC analysis to characterize the bacterial diversity during the fermentation process. GC-clamped amplicons corresponding to a variable region of the bacterial community 16S rDNA were synthesized using polymerase chain reaction (PCR) and then resolved on a base-composition basis using preparative DHPLC. Eluate fractions were collected at random and used as a source of whole community DNA that could be used to determine the bacterial diversity. As a first approach, GC-clamps were removed from the eluted DNA fragments using PCR to avoid the possible bias these clamps could cause during the construction of clone libraries. As a second approach, a clone library of each eluate sample was constructed, preserving the GC-clamps of the DNA fragments. The first approach generated 132 bacterial rDNA sequences with an average size of 200 bp, 45% of which had similarity to unculturable or non-classified bacteria. The second approach produced 194 sequences identified as Proteobacteria (48%), uncultured or non-classified environmental bacteria (40%) and Firmicutes (12%). We detected a remarkably greater bacterial diversity using the first approach than the second approach. The DHPLC-PCR method allowed for the fast and non-laborious detection of a vast bacterial diversity that was associated with cassava fermentation, and we conclude that it is a promising alternative for the characterization of the overall microbial diversity in complex samples.

Cytochemical localization of hydrogen peroxide-generating sites suggests NADPH (nicotinamide adenine dinucleotide 3-phosphate [reduced form]) oxidase expression at the maternal-fetal interface. To explore this possibility, we have characterized the expression and activity of the NADPH oxidase complex in trophoblast cells during the postimplantation period. Implantation sites and ectoplacental cones (EPCs) from 7.5-gestational day embryos from CD1 mice were used as a source for expression analyses of NADPH oxidase catalytic and regulatory subunits. EPCs grown in primary culture were used to investigate the production of superoxide anion through dihydroxyethidium oxidation in confocal microscopy and immunohistochemical assays. NADPH subunits Cybb (gp91phox), Cyba (p22phox), Ncf4 (p40phox), Ncf1 (p47phox), Ncf2 (p67phox), and Rac1 were expressed by trophoblast cells. The fundamental subunits of membrane CYBB and cytosolic NCF2 were markedly upregulated after phorbol-12-myristate-13-acetate (PMA) treatment, as detected by quantitative real-time PCR, Western blotting, and immunohistochemistry. Fluorescence microscopy imaging showed colocalization of cytosolic and plasma membrane NADPH oxidase subunits mainly after PMA treatment, suggesting assembly of the complex after enzyme activation. Cultured EPCs produced superoxide in a NADPH-dependent manner, associating the NADPH oxidase-mediated superoxide production with postimplantation trophoblast physiology. NADPH-oxidase cDNA subunit sequencing showed a high degree of homology between the trophoblast and neutrophil isoforms of the oxidase, emphasizing a putative role for reactive oxygen species production in phagocytic activity and innate immune responses.

Background: The development and progression of cancer depend on its genetic characteristics as well as on the interactions with its microenvironment. Understanding these interactions may contribute to diagnostic and prognostic evaluations and to the development of new cancer therapies. Aiming to investigate potential mechanisms by which the tumor microenvironment might contribute to a cancer phenotype, we evaluated soluble paracrine factors produced by stromal and neoplastic cells which may influence proliferation and gene and protein expression., Methods: The study was carried out on the epithelial cancer cell line (Hep-2) and fibroblasts isolated from a primary oral cancer. We combined a conditioned-medium technique with subtraction hybridization approach, quantitative PCR and proteomics, in order to evaluate gene and protein expression influenced by soluble paracrine factors produced by stromal and neoplastic cells., Results: We observed that conditioned medium from fibroblast cultures (FCM) inhibited proliferation and induced apoptosis in Hep-2 cells. In neoplastic cells, 41 genes and 5 proteins exhibited changes in expression levels in response to FCM and, in fibroblasts, 17 genes and 2 proteins showed down-regulation in response to conditioned medium from Hep-2 cells (HCM). Nine genes were selected and the expression results of 6 down-regulated genes (ARID4A, CALR, GNB2L1, RNF10, SQSTM1, USP9X) were validated by real time PCR., Conclusions: A significant and common denominator in the results was the potential induction of signaling changes associated with immune or inflammatory response in the absence of a specific protein.

Studies on the prevalence of site-specific osteoporosis in patients with different clinical presentations of primary hyperparathyroidism (PHPT) are scarce in the literature. The present study aims to determine the prevalence of cortical osteoporosis in such patients by using 3-site dual-energy X-ray absorptiometry (lumbar spine, femoral neck, 1/3 radius). We studied 49 patients, 12 males and 37 females, with PHPT, who were separated into 3 groups: asymptomatic (AS), renal stone disease (RS), and osteitis fibrosa cystica (OF). Osteoporosis occurred as follows: lumbar spine-48% for AS, 17.6% for RS, and 100% for OF (p=0.0004); femoral neck-20% for AS, 12% for RS, and 85.7% for OF (p=0.0014); 1/3 radius-71% for AS, 53% for RS, and 86% for OF (p=0.2845). Serum calcium, parathyroid hormone, alkaline phosphatase, and beta-carboxy-terminal telopeptide were significantly higher in the OF group. The mean values for 25-hydroxyvitamin D were lower in OF group than AS and RS groups (15.2+/-6.3 ng/mL vs. 22.7+/-11.9 ng/mL and 20.3+/-7.0 ng/mL; p=0.2139). Based on these results, we conclude that the prevalence of osteoporosis is high in all sites studied and is almost universal in the OF patients. The great prevalence of cortical osteoporosis is seen even in young patients with less severe forms of the disease.

Primary hyperparathyroidism often presents as an asymptomatic disorder. In our institution, routine serum calcium measurements have now been used as part of medical examination for 23 years. Out of 124 patients consecutively seen at our institution, 47% presented with no symptoms related to the disease, while 25% presented with severe skeletal involvement and osteitis fibrosa cystica, 25% with renal stone disease without overt bone involvement, and 2% with the typical neuropsychiatric syndrome. This same pattern is seen in the city of São Paulo. In severe disease pathological fractures are frequently seen, especially in long bones of the lower extremities, and also loss of lamina dura of the teeth and salt-and-pepper appearance of the skull. Bone mineral density is extremely low in these patients but usually show remarkable recovery following surgical cure. Serum PTH and bone markers are considerable higher in severely affected patients, who also have a high rate of vitamin D deficiency, and the parathyroid lesion is easier located compared with asymptomatic patients. From pathological specimens 87% had histological confirmation of a single adenoma, 6.4% multiple gland hyperplasia and 3.8% carcinoma.

Paget's disease of bone is a focal disorder of bone remodeling accompanied initially by an increase in bone resorption, followed by a disorganized and excessive formation of bone, leading to pain, fractures and deformities. It exhibits a marked geographical variation in its prevalence. In Brazil it predominantly affects persons of European descent. The majority of the reported cases of the disease in Brazil are from Recife, owing to its peculiar mixed European colonization over approximately four centuries. The etiology is complex and involves both genetic and environmental factors. The disease is often asymptomatic and diagnosis is usually based on biochemical markers of bone turnover, radionuclide bone scan and radiological examination. Bisphosphonates, in particular zoledronic acid, are regarded as the treatment of choice for Paget's disease of bone.

Vitamin D is essential for the maintenance of good health. Its sources can be skin production and diet intake. Most humans depend on sunlight exposure (UVB 290-315 nm) to satisfy their requirements for vitamin D. Solar ultraviolet B photons are absorbed by the skin, leading to transformation of 7-dehydrocholesterol into vitamin D3 (cholecalciferol). Season, latitude, time of day, skin pigmentation, aging, sunscreen use, all influence the cutaneous production of vitamin D3. Vitamin D deficiency not only causes rickets among children but also precipitates and exacerbates osteoporosis among adults and causes the painful bone disease osteomalacia. Vitamin D deficiency has been associated with increased risk for other morbidities such as cardiovascular disease, type 1 and type 2 diabetes mellitus and cancer, especially of the colon and prostate. The prevalence of hypovitaminosis D is considerable even in low latitudes and should be taken into account in the evaluation of postmenopausal and male osteoporosis. Although severe vitamin D deficiency leading to rickets or osteomalacia is rare in Brazil, there is accumulating evidence of the frequent occurrence of subclinical vitamin D deficiency, especially in elderly people.

Objective: To compare vitamin D status represented by serum 25-hydroxyvitamin D (25-OHD) levels in a group of patients with primary hyperparathyroidism, stratified by clinical manifestations., Methods: We studied 22 patients (18 women and 4 men) with primary hyperparathyroidism--5 patients with severe osteitis fibrosa cystica, 10 symptomatic patients with active renal stone disease without overt bone disease, and 7 asymptomatic patients. Bone mineral density (BMD) measurements (T-scores) were done at the lumbar spine, femoral neck, and distal radius, and laboratory data for the three subgroups were analyzed., Results: Although considerably younger than the asymptomatic group, patients with osteitis fibrosa had significantly lower mean serum 25-OHD levels (16.7 +/- 1.1 ng/mL versus 29.9 +/- 2.9 ng/mL; P = 0.003). Moreover, patients with osteitis fibrosa had significantly lower BMD in comparison with patients who had renal stone disease as well as asymptomatic patients for all sites measured. Serum parathyroid hormone levels were significantly higher in patients with osteitis fibrosa than in asymptomatic patients (1,352.8 +/- 297.2 pg/mL versus 145.0 +/- 43.7 pg/mL; P<0.02) as well as in comparison with patients who had renal stone disease (P<0.02). Patients with osteitis fibrosa had urinary N-telopeptide levels that were significantly higher than those in asymptomatic patients (501.5 +/- 201.7 versus 51.3 +/- 6.4 nmol/mmol creatinine; P = 0.02) and those in patients with renal stone disease (P = 0.01)., Conclusion: The findings in this study demonstrate that vitamin D deficiency or insufficiency may contribute to disease severity in primary hyperparathyroidism.