Genomic and physiological characterization of Kitasatospora sp. nov., an actinobacterium with potential for biotechnological application isolated from Cerrado soil.

An Actinobacteria - Kitasatospora sp. K002 - was isolated from the soil of Cerrado, a savanna-like Brazilian biome. Herein, we conducted a phylogenetic, phenotypic and physiological characterization, revealing its potential for biotechnological applications. Kitasatospora sp. K002 is an aerobic, non-motile, Gram-positive bacteria that forms grayish-white mycelium on solid cultures and submerged spores with vegetative mycelia on liquid cultures. The strain showed antibacterial activity against Bacillus subtilis, Pseudomonas aeruginosa and Escherichia coli. Genomic analysis indicated that Kitasatospora xanthocidica JCM 4862 is the closest strain to K002, with a dDDH of 32.8-37.8% and an ANI of 86.86% and the pangenome investigations identified a high number of rare genes. A total of 60 gene clusters of 22 different types were detected by AntiSMASH, and 22 gene clusters showed low similarity (< 10%) with known compounds, which suggests the potential production of novel bioactive compounds. In addition, phylogenetic analysis and morphophysiological characterization clearly distinguished Kitasatospora sp. K002 from other related species. Therefore, we propose that Kitasatospora sp. K002 should be recognized as a new species of the genus Kitasatospora - Kitasatospora brasiliensis sp. nov. (type strains = K002).

Association of coronary calcification with prognosis of Covid-19 patients without known heart disease.

Risk factors that determine the severity of Covid-19 have not been fully elucidated. The aim of this study was to evaluate the role of coronary artery calcification (CAC) as a risk factor for death or mechanical ventilation (MV) of patients without known heart disease infected with Covid-19. We analyzed 283 consecutive in-patients with acute respiratory symptoms with chest computed tomography (chest-CT), without previous heart disease, and criteria for Covid-19 (RT-PCR positive and/or typical clinical and chest-CT findings). CAC was classified by the number of coronary segments affected as absent (0), mild (1-3), and severe calcification (more than 3). The association between CAC, CAC severity, and death or MV due to severe respiratory failure was assessed by logistic regression. The mean age was 58.7±15.7 years and 54.1% were men. Patients with CAC were older, more likely to have hypertension, and less likely to be obese. CAC was present in 75 patients (26.5%), of which 42 had a mild calcification and 33 had severe calcification, and was associated with death (OR=2.35, 95%CI: 1.01-5.48) or MV (OR=2.72, 95%CI: 1.20-6.20) adjusted for multiple confounders, with significant and increased odds ratio for the severe form of CAC (death: OR=3.70, 95%CI: 1.20-11.42; MV: OR=3.30, 95%CI: 1.09-9.95). We concluded that CAC was an independent risk factor for death or MV in Covid-19 patients without previous heart disease, particularly for those with severe calcification. CAC can be easily visualized on common chest-CT, widely used in evaluation of moderate to severe Covid-19.

The applicability of fingernail lead and cadmium levels as subchronic exposure biomarkers for preschool children.

Preschool children are exposed daily to metals in their homes and at daycare centers (DCC). Metal exposure and health effects are associated even at low levels, and children comprise a group of public health concern. Nail metals have been studied for exposure biomonitoring and compared to other biological media. The aim of this study was to explore the applicability of preschool fingernail lead and cadmium as subchronic exposure biomarkers. Nail lead and cadmium levels (NLL and NCL) of 602 preschool children (age: 1-4 years) who attended 21 DCC in São Paulo, Brazil, in 2013 were analyzed. Results were compared against blood lead and cadmium levels (BLL and BCL) found in a previous study. Inductively coupled plasma mass spectrometry (ICP-MS) analyses were performed for both samples. DCC and metal contaminated sites (MCS) were georeferencing. Logistic regression tests were applied to verify associations between nail metal levels and risk factors (sex, age, maternal education, secondary smoking, DCC geographic district, vehicle flow density, relative altitude and distance between DCC and nearest MCS) (p < 0.05). BLL was stratified by exposure level (low: <5 µg.dL-1; high: >13.9 µg.dL-1; medium: ≥5 µg.dL-1 and ≤13.9 µg.dL-1) and also tested the associations in order to verify if nail lead levels are affected by exposure intensity defined by blood metals concentrations. Radius distance and relative altitude of DCC to nearest MCS were associated with high NLL and NCL. Abnormal appearance of nails was associated with high NLL and low NCL. Lead and cadmium exposure magnitude had no significant impact on NLL. NLL should only be used for initial screening, and when financial resources are scarce, especially in areas located near contaminated sites. Preschool children were co-exposed to both lead and cadmium, reinforcing the need for broader studies evaluating exposure to environmental pollutants for more than one chemical element.

Association of coronary calcification with prognosis of Covid-19 patients without known heart disease

Risk factors that determine the severity of Covid-19 have not been fully elucidated. The aim of this study was to evaluate the role of coronary artery calcification (CAC) as a risk factor for death or mechanical ventilation (MV) of patients without known heart disease infected with Covid-19. We analyzed 283 consecutive in-patients with acute respiratory symptoms with chest computed tomography (chest-CT), without previous heart disease, and criteria for Covid-19 (RT-PCR positive and/or typical clinical and chest-CT findings). CAC was classified by the number of coronary segments affected as absent (0), mild (1-3), and severe calcification (more than 3). The association between CAC, CAC severity, and death or MV due to severe respiratory failure was assessed by logistic regression. The mean age was 58.7±15.7 years and 54.1% were men. Patients with CAC were older, more likely to have hypertension, and less likely to be obese. CAC was present in 75 patients (26.5%), of which 42 had a mild calcification and 33 had severe calcification, and was associated with death (OR=2.35, 95%CI: 1.01-5.48) or MV (OR=2.72, 95%CI: 1.20-6.20) adjusted for multiple confounders, with significant and increased odds ratio for the severe form of CAC (death: OR=3.70, 95%CI: 1.20-11.42; MV: OR=3.30, 95%CI: 1.09-9.95). We concluded that CAC was an independent risk factor for death or MV in Covid-19 patients without previous heart disease, particularly for those with severe calcification. CAC can be easily visualized on common chest-CT, widely used in evaluation of moderate to severe Covid-19.

Intra-annual variation in rainfall and its influence of the adults Cyprideis spp (Ostracoda, Crustacea) on a eutrophic estuary (Guanabara Bay, Rio de Janeiro, Brazil) / Variação intra-annual na chuva e a sua influência nos adultos de Cyprideis spp (Ostracoda, Crustacea) em um estuário eutrofizado (Guanabara Bay, Rio de Janeiro, Brasil)

Abstract Spatial and temporal distribution of two species of adult´s ostracods (Cyprideis sp. and Cyprideis salebrosa) were studied as a function of the rainfall patterns in the Guanabara Bay, Rio de Janeiro, Brazil. Samples were taken in ten stations, along six surveys representing three periods (Dry, Early and Late Rainy) for two years. Stations were nested in four areas (Outer, Central, EPA Guapimirim and Impacted). The bottom water (temperature, salinity, dissolved oxygen and oxygen saturation) were measured in each area to characterize the influence of seasonal variations by rainfall. Cyprideis sp. and Cyprideis salebrosa showed patterns distribution to seasonality/surveys (p = 0,002 and p <0,001, respectly). The spatial distribution of Cyprideis sp was significantly different areas studied (p <0.001) indicated well defined areas and distribution in along of the surveys. However, C. salebrosa showed homogeneous distribution in along of the areas within of each survey (p <0.001). Redundancy Analysis (RDA) for the two years evidenced environment preference of the Cyprideis sp. for areas with marine conditions (high influence for channel central) and C. salebrosa for brackish water (high influence of the rivers). This observation reinforces of the existence of areas created by the seasonality of pluviometric regime, a possible dispersion of the adult's ostracods and possibility the use with bioindicators.

Resumo A distribuição espacial e temporal de duas espécies de ostracodes adultos (Cyprideis sp. e Cyprideis salebrosa) foi estudada em função do padrão pluviométrico na Baía de Guanabara, Rio de Janeiro, Brasil. As amostras foram coletadas em dez estações ao longo de seis campanhas, representando três períodos (Seco, Pré e Pós Chuvoso) por dois anos. As estações foram agrupadas em quatro áreas (Externo, Central, EPA Guapimirim e Impactado). A água do fundo (temperatura, salinidade, oxigênio dissolvido e saturação de oxigênio) foi medida em cada área para caracterizar a influência das variações sazonais pela chuva. Cyprideis sp. e Cyprideis salebrosa mostrou distribuição de padrões para sazonalidade/campanhas (p = 0,002 e p <0,001, respctivamente). A distribuição espacial de Cyprideis sp. foi significativamente diferente das áreas estudadas (p <0,001) indicou áreas bem definidas e distribuição ao longo das campanhas. No entanto, C. salebrosa mostrou distribuição homogênea ao longo das áreas dentro de cada campanha (p <0,001). A Análise de Redundância (RDA) para os dois anos evidenciou a preferência ambiental dos Cyprideis sp. para áreas com condições marinhas (alta influência para o canal central) e C. salebrosa para águas salobras (alta influência dos rios). Essa observação reforça a existência de áreas criadas pela sazonalidade do regime pluviométrico, uma possível dispersão dos adultos de ostracodes e a possibilidade do uso como bioindicadores.

How likely is Brazil to achieve its NDC commitments in the energy sector? A review on Brazilian low-carbon energy perspectives.

This paper offers perspectives on the development of low-carbon energy technology in Brazil, pinpointing changes that have occurred since our former publication in 2011. It takes a fresh approach in terms of how likely Brazil will achieve its Nationally Determined Contributions Commitments in the energy sector. Many countries have implemented national climate policies to accomplish their pledged NDC and contribute to the temperature objectives of the Paris Agreement on climate change. Based on official reports and databases of energy development projections in Brazil and the socioeconomic context, we discuss what can be expected for the future of the Brazilian energy sector, the probability of implementing selected technologies, and the prospects of reaching the NDC targets for 2025 and 2030. In addition, this paper provides an overview of the current stage of development of these technologies, main directions, and bottlenecks in Brazil. Analyses have shown that the Brazilian renewable matrix tends to remain significant, driven by the development of solar and mostly small hydroelectric power sources, as well as different types of biomass. In addition, the system will include the replacement of thermoelectric plants powered by diesel and fuel oil by natural gas plants. The prospects for Brazil's official energy plan for 2027 are aligned with the reference technology scenario, which represents the business as usual scenario. Despite this, low-carbon technologies could be implemented far beyond the NDC's goals, given the abundance of renewable natural resources in the country.

Intra-annual variation in rainfall and it's influence of the adult's Cyprideis spp (Ostracoda, Crustacea) on a eutrophic estuary (Guanabara Bay, Rio de Janeiro, Brazil).

Spatial and temporal distribution of two species of adult´s ostracods (Cyprideis sp. and Cyprideis salebrosa) were studied as a function of the rainfall patterns in the Guanabara Bay, Rio de Janeiro, Brazil. Samples were taken in ten stations, along six surveys representing three periods (Dry, Early and Late Rainy) for two years. Stations were nested in four areas (Outer, Central, EPA Guapimirim and Impacted). The bottom water (temperature, salinity, dissolved oxygen and oxygen saturation) were measured in each area to characterize the influence of seasonal variations by rainfall. Cyprideis sp. and Cyprideis salebrosa showed patterns distribution to seasonality/surveys (p = 0,002 and p <0,001, respectly). The spatial distribution of Cyprideis sp was significantly different areas studied (p <0.001) indicated well defined areas and distribution in along of the surveys. However, C. salebrosa showed homogeneous distribution in along of the areas within of each survey (p <0.001). Redundancy Analysis (RDA) for the two years evidenced environment preference of the Cyprideis sp. for areas with marine conditions (high influence for channel central) and C. salebrosa for brackish water (high influence of the rivers). This observation reinforces of the existence of areas created by the seasonality of pluviometric regime, a possible dispersion of the adult's ostracods and possibility the use with bioindicators.

Mirna biogenesis pathway is differentially regulated during adipose derived stromal/stem cell differentiation.

Stromal/stem cell differentiation is controlled by a vast array of regulatory mechanisms. Included within these are methods of mRNA gene regulation that occur at the level of epigenetic, transcriptional, and/or posttranscriptional modifications. Current studies that evaluate the posttranscriptional regulation of mRNA demonstrate microRNAs (miRNAs) as key mediators of stem cell differentiation through the inhibition of mRNA translation. miRNA expression is enhanced during both adipogenic and osteogenic differentiation; however, the mechanism by which miRNA expression is altered during stem cell differentiation is less understood. Here we demonstrate for the first time that adipose-derived stromal/stem cells (ASCs) induced to an adipogenic or osteogenic lineage have differences in strand preference (-3p and -5p) for miRNAs originating from the same primary transcript. Furthermore, evaluation of miRNA expression in ASCs demonstrates alterations in both miRNA strand preference and 5'seed site heterogeneity. Additionally, we show that during stem cell differentiation there are alterations in expression of genes associated with the miRNA biogenesis pathway. Quantitative RT-PCR demonstrated changes in the Argonautes (AGO1-4), Drosha, and Dicer at intervals of ASC adipogenic and osteogenic differentiation compared to untreated ASCs. Specifically, we demonstrated altered expression of the AGOs occurring during both adipogenesis and osteogenesis, with osteogenesis increasing AGO1-4 expression and adipogenesis decreasing AGO1 gene and protein expression. These data demonstrate changes to components of the miRNA biogenesis pathway during stromal/stem cell differentiation. Identifying regulatory mechanisms for miRNA processing during ASC differentiation may lead to novel mechanisms for the manipulation of lineage differentiation of the ASC through the global regulation of miRNA as opposed to singular regulatory mechanisms.

Relation between mast cells concentration and serotonin expression in chagasic megacolon development.

Chagas' disease is still reaching about 10 million people in the world. In South America, one of the most severe forms of this disease is the megacolon, characterized by severe constipation, dilated sigmoid colon and rectum and severe malnutrition. Previous data suggested that mast cells and serotonin (5-hydroxytryptamine [5-HT]) expression could be involved in intestinal homeostasis control, avoiding the chagasic megacolon development. The aim at this study was to characterize the presence of mast cells and expression of serotonin in chagasic patients with and without megacolon and evaluate the relation between mast cells, serotonin and megacolon development. Our results demonstrated that patients without megacolon feature a large amount of serotonin and few mast cells, while patients with megacolon feature low serotonin expression and a lot of mast cells. We believe that serotonin may be involved in the inflammatory process control, triggered by mast cells, and the presence of this substance in large quantities of the intestine could represent a mechanism of megacolon prevention.

Intra-annual variation in rainfall and its influence of the adults Cyprideis spp (Ostracoda, Crustacea) on a eutrophic estuary (Guanabara Bay, Rio de Janeiro, Brazil)

Abstract Spatial and temporal distribution of two species of adult´s ostracods (Cyprideis sp. and Cyprideis salebrosa) were studied as a function of the rainfall patterns in the Guanabara Bay, Rio de Janeiro, Brazil. Samples were taken in ten stations, along six surveys representing three periods (Dry, Early and Late Rainy) for two years. Stations were nested in four areas (Outer, Central, EPA Guapimirim and Impacted). The bottom water (temperature, salinity, dissolved oxygen and oxygen saturation) were measured in each area to characterize the influence of seasonal variations by rainfall. Cyprideis sp. and Cyprideis salebrosa showed patterns distribution to seasonality/surveys (p = 0,002 and p 0,001, respectly). The spatial distribution of Cyprideis sp was significantly different areas studied (p 0.001) indicated well defined areas and distribution in along of the surveys. However, C. salebrosa showed homogeneous distribution in along of the areas within of each survey (p 0.001). Redundancy Analysis (RDA) for the two years evidenced environment preference of the Cyprideis sp. for areas with marine conditions (high influence for channel central) and C. salebrosa for brackish water (high influence of the rivers). This observation reinforces of the existence of areas created by the seasonality of pluviometric regime, a possible dispersion of the adults ostracods and possibility the use with bioindicators.

Resumo A distribuição espacial e temporal de duas espécies de ostracodes adultos (Cyprideis sp. e Cyprideis salebrosa) foi estudada em função do padrão pluviométrico na Baía de Guanabara, Rio de Janeiro, Brasil. As amostras foram coletadas em dez estações ao longo de seis campanhas, representando três períodos (Seco, Pré e Pós Chuvoso) por dois anos. As estações foram agrupadas em quatro áreas (Externo, Central, EPA Guapimirim e Impactado). A água do fundo (temperatura, salinidade, oxigênio dissolvido e saturação de oxigênio) foi medida em cada área para caracterizar a influência das variações sazonais pela chuva. Cyprideis sp. e Cyprideis salebrosa mostrou distribuição de padrões para sazonalidade/campanhas (p = 0,002 e p 0,001, respctivamente). A distribuição espacial de Cyprideis sp. foi significativamente diferente das áreas estudadas (p 0,001) indicou áreas bem definidas e distribuição ao longo das campanhas. No entanto, C. salebrosa mostrou distribuição homogênea ao longo das áreas dentro de cada campanha (p 0,001). A Análise de Redundância (RDA) para os dois anos evidenciou a preferência ambiental dos Cyprideis sp. para áreas com condições marinhas (alta influência para o canal central) e C. salebrosa para águas salobras (alta influência dos rios). Essa observação reforça a existência de áreas criadas pela sazonalidade do regime pluviométrico, uma possível dispersão dos adultos de ostracodes e a possibilidade do uso como bioindicadores.

MicroRNA regulation of stem cell differentiation and diseases of the bone and adipose tissue: Perspectives on miRNA biogenesis and cellular transcriptome.

MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression through targeting and suppression of mRNAs. miRNAs have been under investigation for the past twenty years and there is a large breadth of information on miRNAs in diseases such as cancer and immunology. Only more recently have miRNAs shown promise as a mechanism for intervention with respect to diseases of the bone and adipose tissue. In mesenchymal stem cell (MSC) differentiation, alterations in miRNA expression patterns can differentially promote an osteogenic, adipogenic, or myogenic phenotype. This manuscript reviews the current literature with respect to miRNAs in the context of MSC function with a particular focus on novel avenues for the examination of miRNA associated with bone and adipose tissue biology and disease. Specifically we highlight the need for a greater depth of investigation on MSCs with respect to miRNA biogenesis, processing, strand selection, and heterogeneity. We discuss how these mechanisms facilitate both altered miRNA expression and function.

Crescimento e tolerância à salinidade em três espécies medicinais do gênero Plectranthus expostas a diferentes níveis de radiação / Growth and salinity tolerance in three medicinal species of the genus Plectranthus exposed to different levels of solar radiation

No cultivo de plantas medicinais as condições de salinidade e de luz podem exercer influência no rendimento e na qualidade final da produção de biomassa. O objetivo deste trabalho foi avaliar o efeito de diferentes luminosidades e níveis de salinidade na água de irrigação sobre o crescimento e tolerância de três espécies de plantas do gênero Plectranthus. O ensaio foi conduzido em parcelas subsubdivididas com cinco repetições, sendo as parcelas referente ao fator ambiente (pleno sol e telado), as subparcelas aos níveis de salinidade na água de irrigação - CEa (0,7; 1,9; 3,1; 4,3 e 5,5 dS m-1), e as subsubparcelas às três espécies do gênero Plectranthus (P. amboinicus, P. barbatus e P. grandis). As variáveis de resposta foram: índice relativo de clorofila (IRC), área foliar (AF), razão de área foliar (RAF), área foliar específica (AFE), matéria seca da parte aérea (MSPA), matéria seca das raízes (MSR), matéria seca total (MST), relação MSR/MSPA, grau de tolerância à salinidade, e teor dos íons Na+ e K+. O estresse salino reduziu o crescimento das plantas, sendo as maiores reduções observadas nas plantas expostas a pleno sol. A salinidade influenciou a partição de matéria seca, sendo as raízes mais afetadas do que a parte aérea. Com o aumento da CEa houve aumento expressivo no teor foliar de Na+, enquanto o teor de K+ e o IRC foram reduzidos. Entretanto, o acúmulo de Na+ foi menor em P. grandis. Considerando-se a MST, verificou-se que as três espécies se mostraram moderadamente tolerantes à salinidade de até 3,1 dS m-1, exceto P. grandis cultivada em telado, classificada como tolerante. Em relação ao grau de redução na produção MSPA (parte de interesse comercial), poderia se recomendar o cultivo de P. grandis quando a água de irrigação contiver CEa de até 3,1 dS m-1.

In the cultivation of medicinal plants, salinity and light can affect the yield and quality of biomass. The objective of this study was to evaluate the effect of light and salinity levels of the irrigation water on the growth and salt tolerance of three medicinal plant species of the genus Plectranthus. A completely randomized split-plot design with five repetitions was used. The plots were formed by the environment of cultivation (full sunlight and greenhouse), the subplots by the salinity in the irrigation water - ECw (0.7, 1.9, 3.1, 4.3 and 5.5 dS m-1), and the subsubplots by the three plant species (P. amboinicus, P. barbatus and P. grandis). The following variables were evaluated: relative chlorophyll index (RCI), leaf area (LA), leaf area ratio (LAR), specific leaf area (SLA), shoot dry mass (SDM), root dry mass (RDM), total dry mass (TDM), RDM/SDM ratio, relative salt tolerance, and concentration of Na+ and K+ in leaves. Salinity reduced plant growth, with the greatest reductions observed in plants exposed to full sunlight. Salinity affected the dry matter partitioning, and the roots were more affected than the shoot. With increasing ECw, there was a significant increase in the concentration of Na+ in the leaves, while K+ and RCI decreased. However, the increase in Na+ accumulation was lower in P grandis. The three species were moderately tolerant to salinity up to 3.1 dS m-1, considering the total dry mass production, except the P. grandis grown in greenhouse, classified as tolerant. Regarding the relative reduction in shoot dry mass (part of commercial interest), we could recommend the cultivation of P. grandis when the available irrigation water presents values of ECw up to 3.1 dS m-1.

Morphological aspects of spermatogenic cells at different stagesof sexual maturation in black isogenic C57BL/6/Uni mice / Aspectos morfológicos de células de la espermatogénesis en diferentes etapas de maduración sexual del ratón negro isogénico C57BL/6/Uni

In order to study the morphological changes that occur in cells of the testes of isogenic black mouse C57BL/6/Uni into three periods during spermatogenetic used 15 mice divided into 3 groups of 5 animals with 40, 50 and 60 days of age. The mice were sacrificed and weighed. Testicles were weighed and measured, and histologically processed and stained with HE, PAS and Masson Massom-H and evaluated under light microscopy. It was observed that group I with 40 days of age in the seminiferous tubules had a lumen with sparse small amount of interstitial tubular cells. In the seminiferous epithelium type A spermatogonia, intermediate and B were identified, which occupied the compartment adbasal and intermingled with these cells in spermatocytes I in Pachytene and leptotene was observed, whereas in the adluminal compartment Golgi phase spermatids we observed the presence of acrosomal granule. In group II, the cells of the seminiferous epithelium were developed and it was observed in round spermatids cephalic hood phase plus many elongated spermatids in acrosome phase and Sertoli cells. In group III, 60 days old, it was found that seminiferous epithelium which was of the tubules had elongated spermatids in acrosome phase and maturation, with elongated nuclei and acrosomal system typical of spermiation in the presence of sperm and residual bodies near the tubular lumen. Therefore morphological evolution of germ cell testicular spermatids can be checked and recognized in its four phases: golgi, cap, acrosome and maturation over the age of the animal.

Con el fin de estudiar los cambios morfológicos que ocurren en las células de los testículos del ratón negro isogénico C57BL/6/Uni en tres períodos durante el proceso espermatogenético, se utilizaron 15 ratones divididos en 3 grupos de 5 animales con 40, 50 y 60 días de edad. Los ratones fueron sacrificados y pesados. Luego se pesaron y midieron los testículos, para ser finalmete procesados histológicamente y teñidos con HE, PAS y tricrómico Massom-H y evaluados bajo microscopía de luz. Se observó en el grupo I con 40 días de edad que los túbulos seminíferos tenían un lumen pequeño con escaza cantidad de células tubulares intersticiales. En el epitelio seminífero se identificaron espermatogonias tipo A, intermedio y B, que ocuparon el compartimiento adbasal y entremezcladas con estas células, se observó los espermatocitos I en paquiteno y leptoteno, mientras que en el compartimiento adluminal se observaron espermátidas de fase Golgi en presencia de gránulos acrosomales. En el grupo II, las células del epitelio seminífero se desarrollaron y fue posible observar espermátidas redondas en fase de capuz cefálico además de numerosas espermátidas elongadas en fase acrosómica y células sustentaculares. En el grupo III, de 60 días de edad, se encontró que el epitelio seminífero que revestía los túbulos presentaban espermátidas alargadas en fase acrosómica y de maduración, con sus núcleos alargados y el sistema acrosomal típico de la espermiación, con la presencia de espermatozoides y cuerpos residuales cerca del lumen tubular. Así, se puede comprobar con el paso de la edad del animal, la evolución morfológica de las células germinativas testiculares y reconocer las espermátidas en sus cuatro fases: golgi, capuchón, acrosomal y de maduración.

Morphological aspects of spermatogenic cells at different stages of sexual maturation in black isogenic C57BL/6/Uni Mice / Aspectos morfológicos de las células de la espermatogénesis en diferentesetapas de maduración sexual del ratón negro C57BL/6/Uni

In order to study the morphological changes that occur in cells of the testes of isogenic black mouse C57BL/6/Uni into three periods during spermatogenetic used 15 mice divided into 3 groups of 5 animals with 40, 50 and 60 days of age. The mice were sacrificed and weighed. Then weighed and measured the testicles, to be processed finalmete histologically and stained with HE, PAS and Masson Massom-H and evaluated under light microscopy. Was observed in group I with 40 days of age in the seminiferous tubules had a lumen with sparse small amount of interstitial tubular cells. In the seminiferous epithelium were identified type A spermatogonia, intermediate and B, which occupied the compartment adbasal and intermingled with these cells was observed in spermatocytes I in Pachytene and leptotene, whereas in the adluminal compartment Golgi phase spermatids observed in the presence of acrosomal granule. In group II, the cells of the seminiferous epithelium were developed and it was observed in round spermatids cephalic hood phase plus many elongated spermatids in acrosome phase and Sertoli cells. In Group III, 60 days old, it was found that the seminiferous epithelium which was of the tubules had elongated spermatids in acrosome phase and maturation, with elongated nuclei and acrosomal system typical of spermiation in the presence of sperm and residual bodies near the tubular lumen. So you can check the morphological evolution of germ cell testicular spermatids and recognize its four phases: Golgi, cap, acrosome and maturation over the age of the animal.

Con el fin de estudiar los cambios morfológicos que ocurren en las células del testículo del ratón negro isogénico C57BL/6/Uni en tres períodos diferentes del proceso espermatogenético; fueron utilizados 15 ratones divididos en 3 grupos (n=5) con 40, 50 y 60 días de edad respectivamente. Todos los animales fueron sacrificados y pesados. Posteriormente sus testículos se pesaron, midieron y procesaron histológicamente para HE, PAS y tricrómico Massom-H. Las muestras obtenidas fueron evaluados con microscopía de luz. En el grupo I con 40 días se observaron túbulos seminíferos con un lúmen pequeño y escaza cantidad de células intersticiales. En el epitelio seminífero se identificaron espermatogonias tipo A, intermedio y B, quienes ocuparon el compartimiento basal entremezclándose con espermatocitos I en paquiteno y leptoteno. En el compartimiento adluminal se observaron espermatidas de fase Golgi y presencia de gránulos acrosomales. En el grupo II de 50 días, se observaron células del epitelio seminífero desarrolladas, espermatidas en fase de capuz cefálico, muchas espermatidas elongadas en fase acrosomica y células sustentaculares. En el Grupo III de 60 días se observó el epitelio del túbulo seminífero con espermátidas alargadas en fase acrosómica y de maduración, con núcleos alargados y un sistema acrosomal típico de la espermiación, con presencia de espermatozoides y cuerpos residuales cerca del lúmen tubular. En conclusión se observa la evolución morfológica de las células germinativas testiculares y se reconocen las espermatides en sus cuatro fases: Golgi, capuchón, acrosomal y de maduración en las diferentes edades del animal.

Association between IGF2 and CYP21 gene polymorphisms and characteristics of economic interest in Nellore cattle.

We analyzed two single nucleotide polymorphisms (SNPs) of the IGF2 and CYP21 genes in Nellore cattle participating in the Brazilian Animal Breeding Program. The SNPs were found in exon 6 of the IGF2 (insulin-like growth factor 2) gene (RFLP/MboII) as well as in the promoter region of the CYP21 (steroid 21-hydroxylase) gene (RFLP/HpaII) of these animals. The TC heterozygotes were significantly more frequent than CC and TT homozygotes in the RFLP/MboII polymorphism. The T allele was significantly more frequent than the C allele in RFLP/HpaII polymorphism. This population was found to be in Hardy-Weinberg equilibrium for these SNPs. Association of these polymorphisms with expected progeny differences of reproductive and productive traits was investigated, but proved to be significant only for DP550 (expected progeny differenced for weight at 365 days - IGF2 - RFLP/MboII) and DP450 (expected progeny differenced for weight at 450 days - CYP21 - RFLP/HpaII). This is the first study on the occurrence of these two polymorphisms in this Zebu breed of cattle. A total of 147 Nellore animals participating in the Breeding Program of the Nellore Breed (PMGRN) under the management of the National Association of Breeders and Researchers (ANCP) in the city of Ribeirão Preto were analyzed.

Identification of a DNA methylation point in the promoter region of the bovine CYP21 gene.

The CYP21 (steroid 21-hydroxylase) gene is involved in the synthesis of steroid hormones. Bov-A2 is a retroposon that is common in ruminant genomes. The promoter region of bovine CYP21 contains a short interspersed nucleotide element of Bov-A2, which overlaps a putative Sp1 binding site. We looked for RFLP/HpaII polymorphism in the Bov-A2 element in bovine Zebu breeds by PCR-RFLP, and examined whether polymorphism in this element is associated with methylation. Among DNA samples from 135 Brazilian Zebu breed cattle, we identified an RFLP/HpaII polymorphism (T/C), which, based on a restriction methylation-sensitive assay employing HpaII and isoschizomer MspI enzymes (methylation-sensitive and -non-sensitive enzymes, respectively), appears to be a DNA methylation point. This is the first report of this polymorphism and on DNA methylation in the bovine CYP21 promoter region in Brazilian Zebu cattle.

IGF2/Mboll polymorphism in Gir and Nelore cattle.

The imprinted gene insulin-like growth factor 2 (IGF2) carries out important functions in the development of placental mammals, during the embryonic and fetal stages and is located on bovine chromosome 29. The main aim of this study was to verify the occurrence of a polymorphism (C/T transversion) in exon 6 of the IGF2 gene in Gir and Nelore cattle, Zebu breeds of great economic importance in Brazil for dairy and beef production, respectively. A 193 bp fragment of bovine IGF2 exon 6 was amplified by PCR with specific primers for this region and digested with Mboll enzyme to analyze the polymorphic segment. In a total of 39 Gir animals (29 females and 10 males), the estimated frequencies of the C and T alleles were 0.42 and 0.58, respectively. The genotypic frequencies were 0.10 for CC, 0.26 for TT and 0.64 for CT. For 28 Nelore animals (17 females and 11 males), the estimated frequencies of the C and T alleles were 0.43 and 0.57, respectively. The genotypic frequencies were 0.18 for CC, 0.32 for TT, and 0.50 for CT. This is the first report of the occurrence of this polymorphism in these breeds. The polymorphic site can be a regulatory motif with functional significance to gene regulation of the IGF2 gene. This polymorphism could be used to investigate the allele-specific expression of the IGF2 gene, its epigenetic status, and its role in developmental, growth and reproductive traits.

Neuronal plasticity of the enteric nervous system is correlated with chagasic megacolon development.

Chagas' disease is one of the few functional gastrointestinal disorders for which a causative agent has been identified. However, some pathological aspects of the chagasic megasyndromes are still incompletely understood. Chagasic megacolon is characterized by an inflammatory process, organ dilatation and neuronal reduction in both plexuses of the enteric nervous system (ENS). Although some studies on the ENS in Chagas' disease have been performed, the process of neuronal destruction and neuronal regeneration still remains unclear. Our hypothesis is that the regeneration process of the ENS may be involved with the mechanisms that prevent or retard organ dilatation and chagasic megacolon development. For that reason, we evaluated the neuronal regeneration with the marker GAP-43 in the colon's neuronal plexuses from chagasic patients with megacolon, and from non-infected individuals. Visual examination and quantitative analysis revealed an increased neuronal regeneration process in the dilated portion from chagasic patients when compared with the non-dilated portion and with non-infected individuals. We believe that this increased regeneration can be interpreted as an accentuated neuronal plasticity that may be a response of the ENS to avoid megacolon propagation to the entire organ and maintain the colon functional innervation.

Alternativas para seleção de touros da raça Nelore considerando características múltiplas de importância econômica / Nellore bull selection alternatives using multiple traits of economic importance

Os dados são provenientes de 234 touros da raça Nelore participantes de um teste de progênie, no período de 1996 a 2003. A diferença esperada na progênie (DEP) de sete características: peso aos 120 e 210 dias, efeito materno (DMPP120 e DMPP210), peso e perímetro escrotal aos 365 e 450 dias, efeito direto (DDP365, DDP450, DDPE365 e DDPE450) e idade ao primeiro parto (DDIPP) foi utilizada para classificar os animais em três grupos, assim como identificar quais as características possuíram maior poder discriminatório na formação de cada grupo. Para tanto, foram utilizados procedimentos estatísticos multivariados de análise de agrupamentos k-médias e componentes principais. Os resultados evidenciaram que, dos três grupos formados, dois se destacaram quanto aos valores médios das DEPs. A importância desses dois grupos de touros foi confirmada pela análise de componentes principais, que associou a eles valores superiores de DEPs diretas de peso e perímetro escrotal. A quantidade da variabilidade original retida pelos dois primeiros componentes principais foi de 70,22 por cento. Estes procedimentos mostraram-se eficientes e constituíram importantes ferramentas para classificar touros, discriminar variáveis, bem como resumir informações multivariadas, podendo ser usados como auxílio valioso na seleção de reprodutores para uso nos programas de melhoramento genético.

The data set is from 234 Nelore bulls which participate of a progeny test in the period from 1996 to 2003. The expected progeny diference (EPDs) of seven economic traits, weights at 120 e 210 days of age, maternal effect (DMPP120 and DMPP210), weights and scrotal circumferences at 365 e 450 days of age, direct effects (DDP365, DDP450, DDPE365 and DDPE450), and age at first calving (DDIPP), were used in order to classify these animals in three groups and verify which EPDs showed greatest discriminating power in forming the groups. The statistical applied techniques were: k-means clusters analysis and principal components analysis. From the three groups formed, two of them stood out in relation to values of the EPDs means. Evidence of the importance of these two groups was observed in the principal component analysis that associate to them higher values of direct EPDs of weight and scrotal circumference. The two first principal components accounted for 70.22 percent of the total original variability. Both techniques could be an important tools to sire classification, variable discrimination, as well as to resume multivariate information, and they could be applied to help selection in animal breeding program.