The importance of well protected forests for the conservation genetics of West African colobine monkeys.

In tropical forests, anthropogenic activities are major drivers of the destruction and degradation of natural habitats, causing severe biodiversity loss. African colobine monkeys (Colobinae) are mainly folivore and strictly arboreal primates that require large forests to subsist, being among the most vulnerable of all nonhuman primates. The Western red colobus Piliocolobus badius and the King colobus Colobus polykomos inhabit highly fragmented West African forests, including the Cantanhez Forests National Park (CFNP) in Guinea-Bissau. Both species are also found in the largest and best-preserved West African forest-the Taï National Park (TNP) in Ivory Coast. Colobine monkeys are hunted for bushmeat in both protected areas, but these exhibit contrasting levels of forest fragmentation, thus offering an excellent opportunity to investigate the importance of well-preserved forests for the maintenance of evolutionary potential in these arboreal primates. We estimated genetic diversity, population structure, and demographic history by using microsatellite loci and mitochondrial DNA. We then compared the genetic patterns of the colobines from TNP with the ones previously obtained for CFNP and found contrasting genetic patterns. Contrary to the colobines from CFNP that showed very low genetic diversity and a strong population decline, the populations in TNP still maintain high levels of genetic diversity and we found no clear signal of population decrease in Western red colobus and a limited decrease in King colobus. These results suggest larger and historically more stable populations in TNP compared to CFNP. We cannot exclude the possibility that the demographic effects resulting from the recent increase of bushmeat hunting are not yet detectable in TNP using genetic data. Nevertheless, the fact that the TNP colobus populations are highly genetically diverse and maintain large effective population sizes suggests that well-preserved forests are crucial for the maintenance of populations, species, and probably for the evolutionary potential in colobines.

Human Antigen R (HuR) Facilitates miR-19 Synthesis and Affects Cellular Kinetics in Papillary Thyroid Cancer.

BACKGROUND/AIMS: Pre-mRNA splicing is an essential step in eukaryotic gene expression regulation. Genes are composed of exons that remain in the mature mRNAs and intervening sequences named introns. Splicing is the removal of introns and ligation of exons in a mature transcript. Splice site or spliceosome component mutations can lead to different diseases, including neurodegenerative diseases and several cancer types. HuR is an RNA-binding protein that preferentially binds to U- and AU-rich elements, usually found at the 3' UTRs of some mRNAs. We previously observed HuR specifically associated with spliceosomes assembled on introns containing miR-18a and miR-19a. miR-18a and miR-19a are components of the intronic miR-17-92 cluster, along with other five miRNAs. This cluster has been reported to regulate proliferation, migration, and angiogenesis in cells. In this context, we reasoned HuR could be controlling the splicing and processing of these miRNAs, leading to altered cellular phenotypes. METHODS: We induced HuR overexpression in BCPAP and HEK-293T and analyzed the expression of miRNAs using qPCR, as well as the phenotypic effects in those cells. Cell counting to analyze cell growth was performed after trypan blue staining. Migration and invasion assays were performed using transwell filters and cells were counted after staining with crystal violet. We knocked down HuR using a specific siRNA and analyzed expression of miRNAs by qPCR, as well as cellular kinetics. RESULTS: Our results revealed HuR is associated with miR-19a in BCPAP and HEK-293T cells. Conversely, silencing HuR led to reduced miR-17-5p and miR-19a in BCPAP cells. Our data support that HuR stimulates the expression of miR-19, which is further processed and capable of finding its target sequence in a reporter plasmid. Cells overexpressing HuR showed increased cellular proliferation, migration, and invasion rates. Notably, under the presence of antimiR-19a, BCPAP-HuR cells showed reduced cell growth. Taken together, these results indicate the molecular alterations observed are associated with upregulation of miR-19a, leading to cellular processes involved in cancer development. CONCLUSION: Our findings propose a connection between HuR, miRNA biogenesis and cellular modifications. HuR stimulates miR-19a and miR-19b expression, which leads to up-regulation of cell proliferation, migration and invasion, promoting cancer development.

Shifts of acidogenic bacterial group and biogas production by adding two industrial residues in anaerobic co-digestion with cattle manure.

Anaerobic biodigestion figures as a sustainable alternative to avoid discharge of cattle manure in the environment, which results in biogas and biofertilizer. The anaerobic bioconversion of biomass to methane via anaerobic biodigestion requires a multi-step biological process, including microorganisms with distinct roles. Here, the dynamics of acidogenic bacterial populations by classical microbiology, as well as biogas productivity by gasometer and chromatography, in the anaerobic co-digestion process were studied. This paper presents a performance evaluation of co-digestion systems for biogas production using cattle manure and wastes from the Sewage Treatment Station of a brewery and ricotta cheese whey. The search revealed that the type of substrate added in co-digestion with cattle manure, Carbon/Nitrogen ratio, and Ammonia Nitrogen were the most influential factors that explained many of the variations of the microbiota in the biodigesters fed. This study demonstrated a good potential for the use of ricotta cheese whey in the production of biogas and its further conversion into energy. These findings could provide some fundamental and technical information for the co-treatment of industrial derived wastes in centralized anaerobic biodigestion facilities in a sustainable manner with high process capacity and methane recovery.

Protein disulfide isomerase-mediated transcriptional upregulation of Nox1 contributes to vascular dysfunction in hypertension.

Nox1 signaling is a causal key element in arterial hypertension. Recently, we identified protein disulfide isomerase A1 (PDI) as a novel regulatory protein that regulates Nox1 signaling in VSMCs. Spontaneously hypertensive rats (SHR) have increased levels of PDI in mesenteric resistance arteries compared with Wistar controls; however, its consequences remain unclear. Herein, we investigated the role of PDI in mediating Nox1 transcriptional upregulation and its effects on vascular dysfunction in hypertension. We demonstrate that PDI contributes to the development of hypertension via enhanced transcriptional upregulation of Nox1 in vascular smooth muscle cells (VSMCs). We show for the first time that PDI sulfenylation by hydrogen peroxide contributes to EGFR activation in hypertension via increased shedding of epidermal growth factor-like ligands. PDI also increases intracellular calcium levels, and contractile responses induced by ANG II. PDI silencing or pharmacological inhibition in VSMCs significantly decreases EGFR activation and Nox1 transcription. Overexpression of PDI in VSMCs enhances ANG II-induced EGFR activation and ATF1 translocation to the nucleus. Mechanistically, PDI increases ATF1-induced Nox1 transcription and enhances the contractile responses to ANG II. Herein we show that ATF1 binding to Nox1 transcription putative regulatory regions is augmented by PDI. Altogether, we provide evidence that HB-EGF in SHR resistance vessels promotes the nuclear translocation of ATF1, under the control of PDI, and thereby induces Nox1 gene expression and increases vascular reactivity. Thus, PDI acts as a thiol redox-dependent enhancer of vascular dysfunction in hypertension and could represent a novel therapeutic target for the treatment of this disease.

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells.

MicroRNAs (miRNAs) are short RNA molecules that are widespread in eukaryotes. Most miRNAs are transcribed from introns, and their maturation involves different RNA-binding proteins in the nucleus. Mature miRNAs frequently mediate gene silencing, and this has become an important tool for comprehending post-transcriptional events. Besides that, it can be explored as a promising methodology for gene therapies. However, there is currently a lack of direct methods for assessing miRNA expression in mammalian cell cultures. Here, we describe an efficient and simple method that aids in determining miRNA biogenesis and maturation through confirmation of its interaction with target sequences. Also, this system allows the separation of exogenous miRNA maturation from its endogenous activity using a doxycycline-inducible promoter capable of controlling primary miRNA (pri-miRNA) transcription with high efficiency and low cost. This tool also allows modulation with RNA-binding proteins in a separate plasmid. In addition to its use with a variety of different miRNAs and their respective targets, it can be adapted to different cell lines, provided these are amenable to transfection.

hnRNP A1 and hnRNP C associate with miR-17 and miR-18 in thyroid cancer cells.

Heterogeneous nuclear ribonucleoproteins (hnRNPs) are essential players in the regulation of gene expression. The majority of the twenty different hnRNP proteins act through the modulation of pre-mRNA splicing. Most have been shown to regulate the expression of critical genes for the progression of tumorigenic processes and were also observed to be overexpressed in several types of cancer. Moreover, these proteins were described as essential components for the maturation of some microRNAs (miRNAs). In the human genome, over 70% of miRNAs are transcribed from introns; therefore, we hypothesized that regulatory proteins involved with splicing could be important for their maturation. Increased expression of the miR-17-92 cluster has already been shown to be related to the development of many cancers, such as thyroid, lung, and lymphoma. In this article, we show that overexpression of hnRNP A1 and hnRNP C in BCPAP thyroid cancer cells directly affects the expression of miR-17-92 miRNAs. Both proteins associate with the 5'-end of this cluster, strongly precipitate miRNAs miR-17 and miR-18a and upregulate the expression of miR-92a. Upon overexpression of these hnRNPs, BCPAP cells also show increased proliferation, migration, and invasion rates, suggesting upregulation of these proteins and miRNAs is related to an enhanced tumorigenic phenotype.

Age, but not hand preference, is related to personality traits in common marmosets (Callithrix jacchus).

The proximate mechanisms underlying animal personalities, i.e. consistent inter-individual differences in behaviour, are a matter of discussion. Brain lateralization, expressed as the preferred use of the contralateral limb, has been suggested as one of these mechanisms. In this study, we measured a proxy of brain lateralization in captive common marmosets (N = 28) by testing hand preference in a simple food-reaching task and evaluated personality by coding a wide range of behaviours observed in daily situations. We explored the links between personality and both direction and strength of hand preference, as well as age and sex, using linear models. Principal component analysis revealed that the stable behavioural variables were organized in three personality dimensions: Agreeableness, Extraversion and Neuroticism. Regarding hand preference, 14 individuals were left-handed, seven were right-handed and seven were ambilateral. Contrary to our predictions, we did not find any relationship between personality scores and hand preference or sex. Instead, age was a significant predictor of personality scores, with older individuals being more agreeable and less extraverted. The link between brain lateralization and personality seems to be equivocal and dependent on personality and brain lateralization assessment methods. Further examinations of other proximate mechanisms, such as physiology or (epi)genetics, may elucidate what drives personality variation in common marmosets.

Function Over Mass: A Meta-Analysis on the Importance of Skeletal Muscle Quality in COVID-19 Patients.

COVID-19 caused by SARS-CoV-2 infection is a highly contagious disease affecting both the higher and lower portions of the respiratory tract. This disease reached over 265 million people and has been responsible for over 5.25 million deaths worldwide. Skeletal muscle quality and total mass seem to be predictive of COVID-19 outcome. This systematic review aimed at providing a critical analysis of the studies published so far reporting on skeletal muscle mass in patients with COVID-19, with the intent of examining the eventual association between muscle status and disease severity. A meta-analysis was performed to evaluate whether skeletal muscle quantity, quality and function were related to disease severity. Systematic reviews and meta-analyses were conducted according to the guidelines of the Cochrane Handbook for Systematic Reviews of Interventions and reported according to the guidelines of the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analysis) guide. From a total of 1,056 references found, 480 were selected after removing duplicates. Finally, only 7 met the specified inclusion criteria. The results of this meta-analysis showed that skeletal muscle quality, rather than quantity, was associated with COVID-19 severity, as confirmed by lower skeletal muscle density and lower handgrip strength in patients with severe disease. Muscle function assessment can thus be a valuable tool with prognostic value in COVID-19.

Assessing the Metabolic Impact of Ground Chia Seed in Overweight and Obese Prepubescent Children: Results of a Double-Blind Randomized Clinical Trial.

Childhood obesity is a medical condition of major public health concern. Chia seeds are used to treat certain noncommunicable diseases, and they are rich in omega-3 fatty acids, which contribute to the absorption of vitamins. A randomized double-blind clinical trial of 30 obese children was performed. The sample was composed of prepubertal 5- to 10-year-old children of both sexes with body mass indexes equal to or above the 95th percentile who were recruited through the Pediatric Department of the Faculdade de Medicina do ABC. Blood samples were drawn, the children were weighed and measured, and a 24-h dietary recall was obtained before and after the treatment. Not only were significant differences observed for fibrinogen (P = .011) but a correlation between the changes in markers and the presence of fibers was also observed for two inflammatory parameters: tumor necrosis factor-α (P = .027) and nuclear factor-κß (P = .059). These results indicate that chia seeds may have anti-inflammatory effects related to their fiber content in the context of childhood obesity.

Avaliação das boas práticas de fabricação e da contaminação de preparações cárneas de restaurantes institucionais e comerciais na cidade de Americana, SP / Evaluation of good manufacturing practices and contamination in meat preparations from institutional and commercial restaurants in the city of Americana/SP

Diante do ritmo acelerado da vida contemporânea, observa-se um aumento na tendência dos indivíduos em optar por realizar suas refeições fora de casa. A carne, reconhecida como um componente essencial na alimentação dos brasileiros, está suscetível à contaminação pois apresenta ambiente favorável à proliferação de microrganismos patogênicos. Fazendo-se necessária uma análise de contaminação pós-produção afim de evitar Doenças Transmitidas por Alimentos. No presente estudo objetivouse avaliar as boas práticas de fabricação e contaminação de preparações de carne bovina assada, de restaurantes particulares e institucionalizados no município de Americana-SP. Amostras de carne prontas para o consumo foram obtidas de seis estabelecimentos comerciais e seis institucionais. Durante a coleta, foram verificadas as temperaturas e realizadas análises de conformidades com a RDC n° 275, de 2002. As amostras foram examinadas para detectar a presença ou ausência de E. coli e coliformes termotolerantes a 45° C. Para a análise foi realizada a técnica de tubos múltiplos para quantificar a totalidade dos coliformes. Observou-se que, conforme estipulado pela Resolução n°43 de 2015, nenhuma das amostras oriundas de restaurantes comerciais, e a maioria das provenientes de restaurantes institucionais, atingiram as temperaturas requeridas. No que concerne à identificação de E. coli através de testes microbiológicos, foi constatado que seis amostras de restaurantes comerciais e quatro de restaurantes institucionais testaram positivo para a presença deste microrganismo. Conclui-se que as amostras de restaurantes comerciais apresentaram níveis de contaminação superiores em comparação com as amostras de restaurantes institucionais.

Given the fast-paced rhythm of contemporary life, there is an increase in individuals choosing to have their meals outside the home. Meat, recognized as an essential component in the Brazilian diet, is susceptible to contamination as it provides a favorable environment for the proliferation of pathogenic microorganisms. It is necessary to conduct post-production contamination analysis to prevent Foodborne Diseases. This study aimed to evaluate the good manufacturing practices and contamination of roasted beef preparations from private and institutional restaurants in the city of Americana-SP. Samples of ready-to-eat meat were obtained from six commercial establishments and six institutional ones. During collection, temperatures were checked, and conformity analyses were conducted according to RDC No. 275, 2002. The samples were examined for the presence or absence of E. coli and thermotolerant coliforms at 45°C using the multiple tube technique to quantify the total coliforms. It was observed that, as stipulated by Resolution No. 43, 2015, none of the samples from commercial restaurants and the majority from institutional restaurants reached the required temperatures. Regarding the identification of E. coli through microbiological tests, it was found that six samples from commercial restaurants and four from institutional ones tested positive for the presence of this microorganism. It is concluded that samples from commercial restaurants showed higher contamination levels compared to institutional restaurant samples.

A new model of experimental hemispherotomy in young adult Rattus norvegicus: a neural tract tracing and SPECT in vivo study.

OBJECTIVE: The objective of this study was to describe a new experimental model of hemispherotomy performed on laboratory animals. METHODS: Twenty-six male young adult Wistar rats were distributed into two groups (surgery and control). The nonfluorescent anterograde neurotracer biotinylated dextran amine (BDA; 10,000 MW) was microinjected into the motor cortex area (M1) according to The Rat Brain in Stereotaxic Coordinates atlas to identify pathways and fibers disconnected after the experimental hemispherectomy. SPECT tomographic images of 99mTc hexamethylpropyleneamine oxime were obtained to verify perfusion in functioning areas of the disconnected and intact brain. A reproducible and validated surgical procedure is described in detail, including exact measurements and anatomical relationships. An additional 30 rodents (n = 10 rats per group) were divided into naïve, sham, and hemispherotomy groups and underwent the rotarod test. RESULTS: Cortico-cortical neural pathways were identified crossing the midline and contacting neuronal perikarya in the contralateral brain hemisphere in controls, but not in animals undergoing hemispherotomy. There was an absence of perfusion in the left side of the brain of the animals undergoing hemispherotomy. Motor performance was significantly affected by brain injuries, increasing the number of attempts to maintain balance on the moving cylinder in the rotarod test at 10 and 30 days after the hemispherotomy, with a tendency to minimize the motor performance deficit over time. CONCLUSIONS: The present findings show that the technique reproduced neural disconnection with minimal resection of brain parenchyma in young adult rats, thereby duplicating the hemispherotomy procedures in human patients.

Alterações bioquímicas de plantas e morfológicas de gemas de cafeeiro associadas a eventos do florescimento em resposta a elementos meteorológicos / Biochemical alterations of plants and bud morphology of coffee tree associated to events on flowering in response to meteorological elements

O florescimento do cafeeiro envolve diversos fatores, tanto da planta como do ambiente. A compreensão das interações entre esses fatores pode contribuir para o desenvolvimento de práticas de manejo mais dequadas, principalmente em relação à irrigação. Tais práticas podem vir a promover uma uniformização da floração, conseqüentemente uniformização da maturação dos frutos, minimizando os custos de produção. O objetivo deste trabalho foi avaliar alterações morfológicas de gemas, síntese de prolina e de ácido abscísico em plantas de cafeeiros em resposta a elementos meteorológicos, visando assim contribuir para o conhecimento de fatores que influenciam a uniformização da floração. Foram feitas análises, por meio de técnicas de microscopia eletrônica de varredura, em gemas de ramos plagiotrópicos de primeira ordem, coletadas semanalmente durante o período de novembro/04 a fevereiro/05. No período de julho a setembro de 2005, foram realizadas avaliações em plantas irrigadas e não-irrigadas, de potencial hídrico foliar antes do amanhecer e teor de prolina, em folhas totalmente expandidas, posicionadas em ramos do terço superior da copa. Foi também avaliado o teor de ácido abscísico na seiva do xilema e nas gemas florais. Os resultados mostram modificações na estrutura das gemas associadas a um período de déficit hídrico seguido de precipitação e, conseqüentemente, de menor amplitude térmica. Em relação ao potencial hídrico, há diferenças entre os tratamentos irrigado e não-irrigado e foram observados valores entre -0,3 e -0,8MPa e -0,6 e -1,5MPa respectivamente. Os maiores valores de teores de prolina e de ABA correspondem a períodos de menor oferta de precipitação, entretanto, não há uma relação direta entre a síntese destes compostos e as fases do florescimento avaliadas.

Flowering on coffee plants involves several aspects from the plant as well as from the environment. Understanding these interactions can enhance the knowledge and promote better handling of the crop in field, mainly related to irrigation to promote synchronized flowering and consequently the uniform fruit maturation, lowering the production costs. The aim of the present study was to evaluate bud morphology, proline and abscisic acic biosynthesis on coffee plants under different environmental conditions in order to contribute to the knowledge in factors that influence flowering synchronization. Scanning electron microscopic was performed on buds of plagiotropic shoots weekly collected from November/04 to February/05. Predawn water potential and proline content analysis were carried out from July to September/05 using complete superior third expanded leaves from plagiotropic shoots of non and irrigated plants. Abcisic acid content was also evaluated in xylem sap and on flower buds. The results show alterations on bud morphology associated to the period of water deficit followed by precipitation and also, consequently, lower thermic amplitude. Regarding to the water potential, differences could be observed in the treatments having -0.3 and 0.8MPa for irrigated and -0.6 and -1.5MPa for non-irrigated plants. The highest proline and abcisic acid content are related to dryer period, however there is no direct relationship between these biosynthesis compounds and the different flowering stages evaluated.

As invasöes de áreas públicas municipais em Säo Paulo: uma abordagem sistêmica / The invasions in public areas of Säo Paulo city: a systemic approach

Enfoca a relaçäo entre a propriedade pública da terra e sua apropriaçäo por interesses privados. Articula o fenômeno das invasöes das áreas municipais de Säo Paulo com a história da cidade e do país, quanto à aquisiçäo de terras por particulares. Apresenta um levantamento sistemático do fenômeno das invasöes na última década do século XX, com análises estatísticas quantitativas e espaciais, caracterizando a apropriaçäo da área pública municipal como um fenômeno social generalizado, näo restrito às classes desfavorecidas.