Probabilistic estimation of the source component of seismic hazard in North-Eastern Brazil.

Stable continental regions pose unique challenges for conducting Probabilistic Seismic Hazard Analysis because the earthquake activity driving mechanisms are poorly understood. For instance, the lower seismicity (hence the paucity of data) and the absence of well-defined active fault systems complicate accurately determining seismic source parameters. Northeastern Brazil is a stable continental region exhibiting moderate-size events recorded with significant seismic intensities and provoking the collapse of poorly constructed buildings in the last century. Thus, assessing the seismic hazard is critical for seismic risk mitigation. The seismic hazard depends on three components: source, path, and site, and here, we present the probabilistic seismic hazard analysis of the source component for NE Brazil. Spatial aggregation of earthquake sources outlined four areal seismic zones. A goodness-of-fit test rejected the Gutenberg-Richter model of magnitude frequency distribution in one of the studied seismic zones. For this reason, we estimated the magnitude probability distribution function in that zone using a nonparametric adaptive kernel estimator. In other zones the Gutenberg-Richter magnitude frequency model was applied. In either way of the magnitude probability distribution modelling we considered the upper bound for magnitude equal to 6.6 mR, based on the upper bound of a 95 % confidence interval for the standard normal distribution of palaeoearthquake sizes. Our findings suggests that potentially damaging events are likely to occur, and we cannot neglect chances for the occurrence of earthquakes exceeding 5.2 mR. The calculated mean return periods indicate significantly shorter intervals between consecutive large events than palaeoseismic records.

A Crystallographic Snapshot of SARS-CoV-2 Main Protease Maturation Process.

SARS-CoV-2 is the causative agent of COVID-19. The dimeric form of the viral Mpro is responsible for the cleavage of the viral polyprotein in 11 sites, including its own N and C-terminus. The lack of structural information for intermediary forms of Mpro is a setback for the understanding its self-maturation process. Herein, we used X-ray crystallography combined with biochemical data to characterize multiple forms of SARS-CoV-2 Mpro. For the immature form, we show that extra N-terminal residues caused conformational changes in the positioning of domain-three over the active site, hampering the dimerization and diminishing its activity. We propose that this form preludes the cis and trans-cleavage of N-terminal residues. Using fragment screening, we probe new cavities in this form which can be used to guide therapeutic development. Furthermore, we characterized a serine site-directed mutant of the Mpro bound to its endogenous N and C-terminal residues during dimeric association stage of the maturation process. We suggest this form is a transitional state during the C-terminal trans-cleavage. This data sheds light in the structural modifications of the SARS-CoV-2 main protease during its self-maturation process.

Hepatic injury induced by thioacetamide causes aortic endothelial dysfunction by a cyclooxygenase-dependent mechanism.

Liver cirrhosis is associated with a wide range of cardiovascular abnormalities including hyperdynamic circulation and cirrhotic cardiomyopathy. The pathogenic mechanisms of these cardiovascular changes are multifactorial and include vascular dysregulations. AIM: The present study tested the hypothesis that the systemic vascular hyporesponsiveness in thioacetamide (TAA)-induced liver injury model is dependent on nitric oxide (NO) and cyclooxygenase (COX) derivatives. MAIN METHODS: Wistar rats were treated with TAA for eight weeks to induce liver injury. KEY FINDINGS: The maximal contractile response in concentration-effect curves to phenylephrine was decreased in aorta from TAA-treated rats, but no differences were found in aorta without endothelium, suggesting an endothelium-dependent mechanism in decreased contractile response. There was no difference in the contractile response with and without L-NAME (N(ω)-nitro-l-arginine methyl ester) in rats with liver injury, showing that the TAA treatment impairs NO synthesis. Pre-incubation of the aorta with indomethacin, a COX-inhibitor, normalized the reduced contractile response to phenylephrine in arteries from TAA group. Also, COX-2 and iNOS (inducible nitric oxide syntase) protein expression was increased in aorta from TAA group compared to control group. Animals submitted to TAA treatment had a reduction in systolic blood pressure. Our findings demonstrated that liver injury induced by TAA caused a decrease in aortic contractile response by a COX-dependent mechanism but not by NO release. Also, it was demonstrated an inflammatory process in the aorta of TAA-treated rats by increased expression of COX-2 and iNOS. SIGNIFICANCE: Therefore, there is an essential contribution of COX-2 activation in extra-hepatic vascular dysfunction and inflammation present in cirrhosis induced by TAA.

Cotton genotypes selection through artificial neural networks.

Breeding programs currently use statistical analysis to assist in the identification of superior genotypes at various stages of a cultivar's development. Differently from these analyses, the computational intelligence approach has been little explored in genetic improvement of cotton. Thus, this study was carried out with the objective of presenting the use of artificial neural networks as auxiliary tools in the improvement of the cotton to improve fiber quality. To demonstrate the applicability of this approach, this research was carried out using the evaluation data of 40 genotypes. In order to classify the genotypes for fiber quality, the artificial neural networks were trained with replicate data of 20 genotypes of cotton evaluated in the harvests of 2013/14 and 2014/15, regarding fiber length, uniformity of length, fiber strength, micronaire index, elongation, short fiber index, maturity index, reflectance degree, and fiber quality index. This quality index was estimated by means of a weighted average on the determined score (1 to 5) of each characteristic of the HVI evaluated, according to its industry standards. The artificial neural networks presented a high capacity of correct classification of the 20 selected genotypes based on the fiber quality index, so that when using fiber length associated with the short fiber index, fiber maturation, and micronaire index, the artificial neural networks presented better results than using only fiber length and previous associations. It was also observed that to submit data of means of new genotypes to the neural networks trained with data of repetition, provides better results of classification of the genotypes. When observing the results obtained in the present study, it was verified that the artificial neural networks present great potential to be used in the different stages of a genetic improvement program of the cotton, aiming at the improvement of the fiber quality of the future cultivars.

Haplotypes and polymorphism in the CCR5 gene in sickle cell disease.

Sickle cell disease shows several clinical manifestations in distinct levels of severity. This heterogeneity is due to the haplotype variability associated with the HbS gene, levels of fetal hemoglobin and environmental conditions, which modify the disease expression. Science community believes that the presence of a polymorphism in the CCR5 gene, which is related to chronic inflammatory state, could confer a higher survival rate and a lower number of inflammatory events to these patients since the deletion in CCR5Δ32 would knock out the CCR5 gene. Therefore, this study aimed to identify the haplotypes in ßS and ßC genes, as well as to investigate the presence of the CCR5Δ32 deletion in patients with sickle cell disease. For this purpose, DNA was isolated with the QIAamp DNA Investigator Kit, and PCR was the method chosen to detect the mutant allele CCR5Δ32. The haplotypes in ßS and ßC genes were detected by RFLP with the restriction enzymes XmnI, HindIII, HincII, and HinfI analyzing six polymorphic sites on the ß cluster, succeeded by electrophoresis. The atypical haplotype was the most common (54.3%), followed by Benin (28.6%), Bantu (11.5%), Senegal (2.8%), and Cameroon (2.8%). No patients presented CCR5Δ32 deletion. The increase in the frequency of atypical haplotypes suggests that these patients passed by variation in the genetic pattern from ancestral haplotypes throughout the years.

The molecular motor Myosin Va interacts with the cilia-centrosomal protein RPGRIP1L.

Myosin Va (MyoVa) is an actin-based molecular motor abundantly found at the centrosome. However, the role of MyoVa at this organelle has been elusive due to the lack of evidence on interacting partners or functional data. Herein, we combined yeast two-hybrid screen, biochemical studies and cellular assays to demonstrate that MyoVa interacts with RPGRIP1L, a cilia-centrosomal protein that controls ciliary signaling and positioning. MyoVa binds to the C2 domains of RPGRIP1L via residues located near or in the Rab11a-binding site, a conserved site in the globular tail domain (GTD) from class V myosins. According to proximity ligation assays, MyoVa and RPGRIP1L can interact near the cilium base in ciliated RPE cells. Furthermore, we showed that RPE cells expressing dominant-negative constructs of MyoVa are mostly unciliated, providing the first experimental evidence about a possible link between this molecular motor and cilia-related processes.

Genomic ancestry evaluated by ancestry-informative markers in patients with sickle cell disease.

The ß(s) mutation is responsible for the most aggressive form of sickle cell disease, has a predominantly African origin, and arrived in Brazil through the slave trade. However, the Brazilian population is highly miscegenated, underscoring the importance of ancestry-informative markers (AIMs) for the identification of the genetic structure of a population. In this study, we have estimated the genetic contributions of various ethnicities in individuals with sickle cell disease in the microregion of Jequié, Bahia, in Brazil, by using AIMs, and compared the findings to those from a phenotypic characterization. Eight AIMs were analyzed: AT3 (rs3138521), DRD2 (rs1079598), APO (rs3138522), PV92, Sb19.3 (rs3138524), CKM (rs4884), LPL (rs285), and CCR5Δ32 (rs333). Samples were subjected to polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism. The amplified products were electrophoresed on agarose gels, and the data were statistically analyzed using Genepop, FSTAT 2.9, and Admix3. Phenotypic classification showed a high frequency of mulattos  (85%) in the Brazilian population; however, ancestry-informative markers indicated that 44, 42, and 11% of the population had European, African, and native American ancestries, respectively. The phenotypic classification is justified as a complementary method for the characterization of the genetic ancestry in patients with sickle cell disease, as it confirms the molecular findings regarding ancestry.

Classification of different degrees of adiposity in sedentary rats.

In experimental studies, several parameters, such as body weight, body mass index, adiposity index, and dual-energy X-ray absorptiometry, have commonly been used to demonstrate increased adiposity and investigate the mechanisms underlying obesity and sedentary lifestyles. However, these investigations have not classified the degree of adiposity nor defined adiposity categories for rats, such as normal, overweight, and obese. The aim of the study was to characterize the degree of adiposity in rats fed a high-fat diet using cluster analysis and to create adiposity intervals in an experimental model of obesity. Thirty-day-old male Wistar rats were fed a normal (n=41) or a high-fat (n=43) diet for 15 weeks. Obesity was defined based on the adiposity index; and the degree of adiposity was evaluated using cluster analysis. Cluster analysis allowed the rats to be classified into two groups (overweight and obese). The obese group displayed significantly higher total body fat and a higher adiposity index compared with those of the overweight group. No differences in systolic blood pressure or nonesterified fatty acid, glucose, total cholesterol, or triglyceride levels were observed between the obese and overweight groups. The adiposity index of the obese group was positively correlated with final body weight, total body fat, and leptin levels. Despite the classification of sedentary rats into overweight and obese groups, it was not possible to identify differences in the comorbidities between the two groups.

Classification of different degrees of adiposity in sedentary rats

In experimental studies, several parameters, such as body weight, body mass index, adiposity index, and dual-energy X-ray absorptiometry, have commonly been used to demonstrate increased adiposity and investigate the mechanisms underlying obesity and sedentary lifestyles. However, these investigations have not classified the degree of adiposity nor defined adiposity categories for rats, such as normal, overweight, and obese. The aim of the study was to characterize the degree of adiposity in rats fed a high-fat diet using cluster analysis and to create adiposity intervals in an experimental model of obesity. Thirty-day-old male Wistar rats were fed a normal (n=41) or a high-fat (n=43) diet for 15 weeks. Obesity was defined based on the adiposity index; and the degree of adiposity was evaluated using cluster analysis. Cluster analysis allowed the rats to be classified into two groups (overweight and obese). The obese group displayed significantly higher total body fat and a higher adiposity index compared with those of the overweight group. No differences in systolic blood pressure or nonesterified fatty acid, glucose, total cholesterol, or triglyceride levels were observed between the obese and overweight groups. The adiposity index of the obese group was positively correlated with final body weight, total body fat, and leptin levels. Despite the classification of sedentary rats into overweight and obese groups, it was not possible to identify differences in the comorbidities between the two groups.

Impairment of Interleukin-17A Expression in Canine Visceral Leishmaniosis is Correlated with Reduced Interferon-γ and Inducible Nitric Oxide Synthase Expression.

Dogs are the primary urban reservoir of Leishmania infantum and play a crucial role in the transmission of this parasite to man via sandflies. The spleen and liver are the main target organs of L. infantum infection, but few studies have evaluated the immune response to this infection in the canine liver. To identify the immunological mediators involved in resistance and/or susceptibility to canine visceral leishmaniosis (CVL), we selected 21 dogs naturally infected by L. infantum and classified as asymptomatic or symptomatic. Immunological parameters were analysed and correlations with clinical signs were determined. Symptomatic dogs showed higher numbers of parasites and less leucocyte infiltration in the liver compared with asymptomatic dogs. The progression of this disease was characterized not only by the down regulation of T helper (Th) 1-related cytokines, such as interferon (IFN)-γ and tumour necrosis factor (TNF)-α, but also by the down regulation of genes encoding interleukin (IL)-17A, inducible nitric oxide synthase (iNOS) and IL-10 in the spleen and liver in symptomatic dogs compared with asymptomatic dogs. Importantly, IL-17A gene transcription level was positively correlated with mRNA expression for iNOS and IFN-γ. Th1- and Th17-related cytokines therefore appear to play a role in restricting parasite growth via iNOS activation and decrease susceptibility of dogs to CVL.

Lycopene-rich tomato oleoresin modulates plasma adiponectin concentration and mRNA levels of adiponectin, SIRT1, and FoxO1 in adipose tissue of obese rats.

AIM: To investigate whether lycopene can modulate adiponectin levels and SIRT1 and FoxO1 gene expression in the adipose tissue of diet-induced obese rats. METHODS: Male Wistar rats were first fed with hypercaloric diet (HD, n = 12) for 6 weeks, and afterward, these rats were randomly assigned to receive HD (n = 6) or HD with lycopene-rich tomato oleoresin (equivalent to lycopene 10 mg/kg body weight (BW)/day, HD + L, n = 6) by gavage for additional 6 weeks. Plasma lycopene and adiponectin levels were analyzed by high-performance liquid chromatography and immunoassay, respectively. The messenger RNA (mRNA) expressions of adiponectin, Sirtuin 1 (SIRT1), Forkhead box O 1 (FoxO1), fatty acid translocase/cluster of differentiation 36 (FAT/CD36), and PPARγ in adipose tissues were determined by quantitative polymerase chain reaction. RESULTS: Lycopene was detected in the plasma of rats in HD + L group but not in the HD group. Although both BW and adiposity were not different between the two groups, there was a significant increase in both plasma concentration and mRNA expression of adiponectin in the adipose tissue of the HD + L group. In addition, the lycopene supplementation upregulated mRNA expressions of SIRT1, FoxO1, and FAT/CD36 but downregulated PPARγ in adipose tissue of obese rats. CONCLUSION: These data suggest that lycopene, in the concentration used, is not toxic and also its health benefits in adipose tissue may play a role against obesity-related complications.

Lycopene supplementation reduces TNF-α via RAGE in the kidney of obese rats.

BACKGROUND: The kidney is a target organ for injuries caused by advanced glycation end products (AGEs) in obesity. The receptor of AGEs (RAGE) is proinflammatory and appears to have a role in the pathogenesis of renal disease due to obesity. OBJECTIVE: The aim was to verify the effect of obesity on renal damage and the effect of lycopene on these complicationsDesign and Methods:Male Wistar rats were randomly assigned to receive a control diet (C, n=7) or a high-fat diet plus sucrose (HD+S, n=14) for 6 weeks. After this period, the HD+S animals were randomized into two groups: HD+S (n=7) and HD+S supplemented with lycopene (HD+S+L, n=7). The animals received maize oil (C and HD+S) or lycopene (HD+S+L) for a 6-week period. RESULTS: The HD+S and HD+S+L animals demonstrated insulin resistance (OGTT glucose after 150 min; C: 117.6±3.9

Initiation and s uppression of sperm motility is osmolality dependent in two South American fish species: streaked prochilod (Prochilodus lineatus) and piracanjuba(Brycon orbignyanus)

Solutions to induce or suppress the initiation of sperm motility in fish ha ve been used to improve reproductive success during artificial fertilization and preservation techniques . The aim of the present study was to evaluate the effects of three solutions (NaCl, glucose , and BTS™) - each prepared with 10 different osmolalities - on the initiation and suppression of fresh sperm motility in Prochilodus lineat us and Brycon orbignyanus . Sperm was diluted in each of the 30 solution s and immediately observed under a light microscope to determine which solution s trigger ed or suppress ed the initiation of sperm motility. When present, motility rate ( % motile sperm ) w as determined at 0, 30 , and 120 s post - activation and the motility quality score ranging from 0 ( no movement ) to 5 ( rapidly swimming sperm) was determined at 0 and 30 s post - activation . Osmolality , but not solution composition , significantly affected both motility rate and quality score . Solutions at osmolali ties up to 270 mOsm/kg in P. lineatus and up to 180 mOsm/kg in B. orbignyanus induced motility in at least 60% of sperm , with a minimum quality score of 3.0 , and were therefore classified as activating agents. The greatest motility at 0 , 30 , and 120 s post - activation was observed with solutions ranging from 135 to 225 mOsm/kg for P. lineatus and at 135 mOsm/kg for B. orbignyanus . On the other hand, solutions ranging from 360 to 450 mOsm/kg in P. lineatus and 270 to 450 mOsm/kg in B. orbignyanus suppressed motility in at least 95% of sperm and were classified as immobilizing media . The osmolality of the surrounding medium is the key factor in the initiation or suppression of sperm motility in P. lineatus a nd B. orbignyanu.

Efeito materno e paterno sobre as taxas de fertilização e eclosão em curimba (Prochilodus lineatus) / Maternal and paternal effect on fertilization and haching rate in curimba (Prochilodus lineatus)

Avaliou-se o quanto fêmeas e machos contribuem para a variação total das taxas de fertilização e de eclosão em curimba (Prochilodus lineatus). Utilizou-se sêmen criopreservado proveniente de cinco machos para fertilizar ovócitos de seis fêmeas em um esquema fatorial cruzado 5x6, totalizando 30 famílias. Além das características reprodutivas dos machos e fêmeas, foram avaliadas as taxas de fertilização e eclosão para cômputo dos efeitos materno e paterno. Os componentes da variância foram estimados por meio da máxima verossimilhança restrita, sendo construídos intervalos Highest Posterior Density (HPD) para cada componente. Verificou-se que as fêmeas contribuíram muito mais para a variação total em relação aos machos para as taxas de fertilização e eclosão. Para a taxa de fertilização, as fêmeas contribuíram com 26,3% da variação total e os machos com 8,9%. Em relação à taxa de eclosão, as fêmeas contribuíram com 11,9% e os machos com 1,6%. Concluiu-se que houve efeito materno sobre as taxas de fertilização e eclosão e que o efeito paterno avaliado individualmente foi pouco expressivo ou até mesmo insignificante.

The aim of this study was to evaluate how much females and males contribute to the total variation of reproductive traits such as fertilization and hatching rate in curimba Prochilodus lineatus. Cryopreserved semen from five males was used to fertilize eggs from six females in a cross-factorial 5x6, totaling 30 families. In addition to the reproductive characteristics of males and females, fertilization and hatching rates were evaluated for computation of maternal and paternal effects. The variance components were estimated by restricted maximum likelihood, and the Highest Posterior Density (HPD) intervals were estimated for each component. The female contributed more to the total variation than males for the fertilization and hatching rates. The female contributed 26.3% of the total variation in the fertilization rate against 8.9% of males. Regarding the hatching rate, the female contributed 11.9% versus 1.6% of males. Thus, there is maternal effect on rates of fertilization and hatching, and the paternal effect assessed individually was lackluster or even negligible.

Effects of extenders, cryoprotectants and freezing methods on sperm quality of the threatened Brazilian freshwater fish pirapitinga-do-sul Brycon opalinus (Characiformes).

The objective was to develop a suitable freezing method to cryopreserve Brycon opalinus (Characiformes) sperm. Extenders (NaCl and glucose at 325 and 365 mOsm/kg), cryoprotectants (dimethyl sulfoxide=dimethyl sulfoxide (DMSO) and methyl glycol=methyl glycol (MG)), equilibration times (15 and 30 min), thawing temperatures (30 and 60 °C), and straw sizes (0.5 and 4.0 mL) were tested. Sperm were frozen in a liquid nitrogen vapor vessel at -170 °C and subsequently stored in liquid nitrogen. Post-thaw sperm quality was always evaluated in terms of motility (expressed as percentage of motile sperm), duration of motility and vitality (eosin-nigrosin staining, expressed as percentage of intact sperm). The best freezing method was also tested for fertility and hatching (expressed as the percentage of fertilized eggs). Post-thaw sperm quality was highest when sperm were cryopreserved in Glucose 365 mOsm/kg and MG, after a 30-min equilibration and thawed at 60 °C for 8 s, of regardless straw size: 74±7% motile sperm, 47±4 s of motility duration, 69±3% intact sperm, 64±4% fertilization and 63±3% hatching. The freezing method developed in the present study was efficient and can be used to maximize larvae production for both aquaculture purposes and for conservational programs, since B. opalinus is a threatened species.

The use of medicinal plants by an indigenous Pataxó community in NE Brazil / O uso de plantas medicinais por uma comunidade indígena Pataxó no NE do Brasil

We identified and classified 48 medicinal plants used by the Pataxó Indians in south Bahia, Brazil. The location is an ecologically threatened area designated by UNESCO as a World Heritage Site. The aim of this paper is to document phytotherapeutic practices in the indigenous community of Mata Medonha. We conducted interviews with the 25 families present at the area. Of the 48 medicinal species identified, only 14 (29%) had been examined for mechanism of action or isolation of biochemical compounds, according to bibliographic research. The plants were deposited at the Alexandre Leal Costa Herbarium of the Federal University of Bahia. We gathered information about the preparation and uses of the plants. The species are used for a variety of maladies, including flu, congestion, bronchitis and headaches, pain, snake bites, and some were only used for women's disorders. The Pataxó ethnopharmacological knowledge is under pressure from the economic outmigration of the community and threats to the biodiversity from logging, mining, and tourism. The plants studied here include important drug candidates. Additional research on the molecular aspects of the species cited should be performed.

Foram identificadas e classificadas 48 plantas medicinais usadas por Índios Pataxós no sul da Bahia. A região é área de risco ecológico designada pela UNESCO como Sítio do Patrimônio Mundial. O objetivo deste estudo foi de documentar as práticas fitoterápicas na comunidade indígena da Mata Medonha. Foram conduzidas entrevistas com as 25 famílias presentes no local. Dentre as 48 espécies identificadas, apenas 14 (29%) foram avaliadas quanto ao mecanismo de ação ou isolamento de compostos químicos, de acordo com levantamento bibliográfico. As plantas foram depositadas no Herbário Alexandre Leal Costa da Universidade Federal da Bahia. Foram registradas informações sobre o preparo e uso das plantas citadas. As espécies são usadas para uma variedade de doenças incluindo gripe, congestão, bronquite, dores em geral, picadas de cobra e algumas apenas para doenças femininas. O conhecimento etnofarmacológico Pataxó está sobre pressão pelas migrações para fora da comuindade e ameaças à biodiversidade por desmatamento, mineração e turismo. As plantas estudadas incluem importantes candidatos a fármacos. Pesquisas adicionais sobre os aspectos moleculares devem ser realizadas.

Supraphysiological triiodothyronine doses diminish leptin and adiponectin gene expression, but do not alter resistin expression in calorie restricted obese rats.

Thyroid hormones regulate energy balance and act on adipokines. However, while it is unclear what the effects are of calorie restriction and high doses of triiodothyronine (T(3)) on adipokines in obesity, thyroid hormones are illicitly administered in isolation or in association with a hypocaloric diet as an obesity treatment. The present study determined the effect of T(3) on serum concentrations and gene expression of the adipokines leptin, resistin, and adiponectin in calorie-restricted obese rats. Male Wistar rats received a hypercaloric diet for 20 weeks followed by calorie restriction for 8 weeks. The animals were then randomly divided into 3 groups: calorie restriction (OR), OR with 5 µg of T(3)/100 g BW (RS1), and OR with 25 µg of T(3)/100 g BW (RS2) for 2 weeks. Blood and adipose tissue samples were collected for biochemical, hormonal, and gene expression analyses. Serum concentrations of leptin (OR: 3.7±0.6, RS1: 3.8±1, RS2 0.2±0.07 ng/dl) and resistin (OR: 2.5±0.6, RS1: 2.5±0.5, RS2 1.6±0.3 ng/dl) were diminished at the higher dose, while serum adiponectin (OR: 31±7, RS1: 24±5, RS2 26±7 ng/dl) levels were lower in the low dose group. Administration of T(3) reduced leptin gene expression (OR: 0.91±0.1, RS1: 0.95±0.1, RS2 0.22±0.1) only at the higher dose, resistin expression (OR: 1.06±0.2, RS1: 1.04±0.1, RS2 0.88±0.2) was not influenced by T(3) treatment, and adiponectin expression (OR: 1.55±0.5, RS1: 0.95±0.15, RS2 0.97±0.13) was diminished independent of the T(3) dose. These results indicate that T(3), directly or indirectly, inhibits the expression of leptin and adiponectin in calorie restricted obese animals.

Inhibitor of PI3Kγ ameliorates TNBS-induced colitis in mice by affecting the functional activity of CD4+CD25+FoxP3+ regulatory T cells.

BACKGROUND AND PURPOSE: Phosphoinositide 3-kinase-γ (PI3Kγ) is implicated in many pathophysiological conditions, and recent evidence has suggested its involvement in colitis. In the present study, we investigated the effects of AS605240, a relatively selective PI3Kγ inhibitor, in experimental colitis and its underlying mechanisms. EXPERIMENTAL APPROACH: Acute colitis was induced in mice by treatment with trinitrobenzene sulphonic acid (TNBS), and the effect of AS605240 on colonic injury was assessed. Pro-inflammatory mediators and cytokines were measured by immunohistochemistry, elisa, real time-polymerase chain reaction and flow cytometry. KEY RESULTS: Oral administration of AS605240 significantly attenuated TNBS-induced acute colitis and diminished the expression of matrix metalloproteinase-9 and vascular endothelial growth factor. The colonic levels and expression of IL-1ß, CXCL-1/KC, MIP-2 and TNF-α were also reduced following therapeutic treatment with AS605240. Moreover, AS605240 reduced MIP-2 levels in a culture of neutrophils stimulated with lipopolysaccharide. The mechanisms underlying these actions of AS605240 are related to nuclear factor-κ (NF-κB) inhibition. Importantly, the PI3Kγ inhibitor also up-regulated IL-10, CD25 and FoxP3 expression. In addition, a significant increase in CD25 and FoxP3 expression was found in isolated lamina propria CD4+ T cells of AS605240-treated mice. The effect of AS605240 on Treg induction was further confirmed by showing that concomitant in vivo blockade of IL-10R significantly attenuated its therapeutic activity. CONCLUSIONS AND IMPLICATIONS: These results suggest that AS605240 protects mice against TNBS-induced colitis by inhibiting multiple inflammatory components through the NF-κB pathway while simultaneously inducing an increase in the functional activity of CD4+CD25+ Treg. Thus, AS605240 may offer a promising new therapeutic strategy for the treatment of inflammatory bowel diseases.

Motility and fertility of the subtropical freshwater fish streaked prochilod (Prochilodus lineatus) sperm cryopreserved in powdered coconut water.

Streaked prochilod (Prochilodus lineatus) is a freshwater fish inhabiting many South American rivers. The objective was to determine the effectiveness of coconut water (ACP), combined with methylglycol, as a freezing medium for streaked prochilod sperm. A secondary objective was to compare a computer-assisted sperm analyzer (CASA) system versus subjective microscropic examination as a means of assessing sperm motility. As a control, glucose and methylglycol was used, according to our previous study. Sperm diluted in each medium was loaded into 0.5 mL straws, frozen in liquid nitrogen vapor (in a dry shipper), and stored in liquid nitrogen (-196 degrees C). Half of the samples were evaluated for sperm motility, both subjectively and with CASA; the remainder were evaluated for fertility. There was no difference (P > 0.05) between subjective or CASA assessment of post-thaw sperm motility. Although sperm motility was higher in sperm cryopreserved in ACP (85%) than in glucose (75%), cryopreservation in either extender yielded similar fertilization rates (46-48%) and sperm velocities. There were positive correlations (r = 0.56-0.8) between all sperm velocities and fertilization rate. In conclusion, streaked prochilod sperm cryopreserved in glucose or ACP and methylglycol was fertile, and thus could be used for research or commercial settings. Furthermore, although the CASA system provided objective data regarding sperm motility, in the present study, subjective evaluation of sperm motility was practical and a good indication of sperm quality; it could readily be done by well-trained personnel under field or laboratory conditions.

Out-of-season sperm cryopreserved in different media of the Amazonian freshwater fish pirapitinga (Piaractus brachypomus).

The pirapitinga (Piaractus brachypomus) is a freshwater fish that inhabits the Amazon and Orinoco River basins. The use of cryopreserved sperm has been considered to facilitate procedures during the artificial reproduction. The aim of the present study was to develop a freezing protocol for pirapitinga sperm collected outside the spawning season. Sperm samples were diluted in four freezing media prepared by a combination of two extenders (glucose and BTS-Beltsville Thawing Solution) and two cryoprotectant agents (DMSO and methylglycol) loaded into 0.5-mL straws, frozen in a nitrogen-vapor shipping dewar (dry-shipper) and stored in liquid nitrogen at -196 degrees C. Post-thaw sperm motility was evaluated both subjectively using a light microscope and by a computer-assisted sperm analyzer (CASA). Curvilinear, average path and straight-line velocities were also determined. There were no differences (P>0.05) in post-thaw sperm motility between evaluations performed subjectively and using the CASA. Sperm samples cryopreserved in glucose-methylglycol yielded the greatest post-thaw sperm motility (81%) and fastest sperm velocities when compared to the samples frozen in the other three media (P<0.05). Out-of-season sperm cryopreserved in glucose and methylglycol under the conditions described above is of high quality and can therefore be used to facilitate artificial reproduction procedures, as only females will need handling for hormonal induction and gamete collection during the spawning season. Although the CASA system provides precise data on sperm motility, the subjective evaluation is practical and can be conducted by well-trained personnel at commercial fish farms as an acceptable evaluation of sperm quality.