Effect of retinol dehydrogenase gene transfer in a novel rat model of Stargardt disease.

Dysfunction of the ATPase-binding Cassette Transporter protein (ABCA4) can lead to early onset macular degeneration, in particular to Stargardt disease. To enable translational research into this form of blindness, we evaluated the effect of Cas9-induced disruptions of the ABCA4 gene to potentially generate new transgenic rat models of the disease. We show that deletion of the short exon preceding the second nucleotide-binding domain is sufficient to drastically knock down protein levels and results in accumulation of retinoid dimers similar to that associated with Stargardt disease. Overexpression of the retinol dehydrogenase enzymes RDH8 and RDH12 can to a limited extent offset the increase in the bisretinoid levels in the Abca4Ex42-/ - KO rats possibly by restricting the time window in which retinal can dimerize before being reduced to retinol. However, in vivo imaging shows that overexpression of RDH8 can induce retinal degeneration. This may be due to the depletion in the outer segment of the cofactor NADPH, needed for RDH function. The translational potential of RDH therapy as well as other Stargardt disease therapies can be tested using the Abca4 knockdown rat model.

Microencapsulated and uncoated butyric acid as alternative additives to the regeneration of intestinal mucosa in broilers challenged with Eimeria spp.

1. The effect of microencapsulated and uncoated butyric acid as an alternative to antibiotics on performance, intestinal morphology and regeneration of intestinal mucosa was studied in birds experimentally infected with Eimeria spp. 1 to 42 d-old.2. A total of 1,320 male Cobb® broiler chicks were allocated to one of five treatments in a completely randomised design, comprising a negative control, uncoated butyric acid (UA), microencapsulated butyric acid (MA), combined U + M butyric acid and a positive control (antibiotic+anticoccidial) in six replications. At 16 d-old, the birds were inoculated orally with 0.5 ml of a solution containing an Eimeria spp. pool.3. At 21 d of age, the birds receiving butyric acid alone had higher body weight gain (BWG) and feed intake (FI) compared to those supplemented with the blend of acids. For the total rearing period, in all variables, the positive control performed best (P < 0.001).4. At 14 d of age, birds that received diets containing UA had a deeper crypt depth in the jejunum than those fed diets containing microencapsulated acid (P = 0.0194). At 21 d of age, the birds fed the acids had higher villi (P = 0.0058) in the duodenum, compared to the negative control group.5. Supplementation with microencapsulated acid contributed to the intestinal health and recovery of post-challenge birds, but did not result in improvements in performance.

Agaricus subrufescens and Pleurotus ostreatus mushrooms as alternative additives to antibiotics in diets for broilers challenged with Eimeria spp.

1. The effect of A. subrufescens and P. ostreatus mushrooms as an alternative to antibiotics (avilamycin or monensin sodium) on performance, intestinal morphometry, immunity, and biochemical profile of broilers challenged with Eimeria spp. was studied from 1 to 42 d old. A total of 900 male Cobb® broiler chicks were distributed, according to a completely randomised design, into five treatments with six replicates each.2. The treatments consisted of: negative control (NC) - basal diet (BD) with no anticoccidial or antibiotic (non-challenged birds); negative control challenged (NCC) - NC fed to Eimeria spp. challenged birds; BD with 0.2% A. subrufescens inclusion for challenged birds (As), BD with 0.2% P. ostreatus inclusion for challenged birds (Po); and a positive control - BD with anticoccidial and antibiotic inclusion for challenged birds (ATB).3. At 11 d.o., the birds were each inoculated orally with 1 ml solution containing 2 × 105 sporulated oocysts/ml Eimeria acervulina and 2 × 104 sporulated oocysts/ml E. maxima and E. tenella.4. Birds subjected to Eimeria spp. challenge up to 21 d of age had greater crypt depth, indicating that the presence of undesirable microorganisms had an effect on cell proliferation.5. At 21 d old, the birds receiving ATB had higher average weight gain (AWG), feed intake (AFI), and feed conversion ratio (FCR) compared to those fed diets supplemented with mushrooms (As or Po). For the total rearing period (42 days), the birds that received ATB had higher AWG and AFI (P < 0.001) compared to those that received As or Po diets. Feeding avilamycin did not affect (P = 0.0676) FCR compared to the As or Po diet groups.6. From the morphometric and blood analyses there were no differences between broilers fed ATB, Po or As diets in either rearing periods. However, Po and As supplementation lowered blood triglyceride levels. At 21d there was a difference (P < 0.05) for MCV and haemoglobin, in which the mushrooms were similar to the antibiotic. At 42 d, there was a difference (P < 0.05) in haematocrit, erythrocyte, MCV, H: L, protein and albumin variables, in which the use of mushrooms was similar to the positive control, demonstrating that both (mushrooms and antibiotics) promoted a certain improvement in the health of the chickens.7. A. subrufescens and P. ostreatus can be used in broiler diets without compromising intestinal or haematological status, however, these ingredients did not result in improvements in performance.

Long-term expression of melanopsin and channelrhodopsin causes no gross alterations in the dystrophic dog retina.

Several preclinical studies have investigated the potential of algal channelrhodopsin and human melanopsin as optogenetic tools for vision restoration. In the present study, we assessed the potentially deleterious effects of long-term expression of these optogenes on the diseased retina in a large animal model of retinal degeneration, the RPE65-deficient Briard dog model of Leber congenital amaurosis. Intravitreal injection of adeno-associated virus vectors expressing channelrhodopsin and melanopsin had no effect on retinal thickness over a 16-month period post injection. Our data support the safety of the optogenetic approach for the treatment of blindness.

The Joule heating problem in silver nanowire transparent electrodes.

Silver nanowire transparent electrodes have shown considerable potential to replace conventional transparent conductive materials. However, in this report we show that Joule heating is a unique and serious problem with these electrodes. When conducting current densities encountered in organic solar cells, the average surface temperature of indium tin oxide (ITO) and silver nanowire electrodes, both with sheet resistances of 60 ohms/square, remains below 35 °C. However, in contrast to ITO, the temperature in the nanowire electrode is very non-uniform, with some localized points reaching temperatures above 250 °C. These hotspots accelerate nanowire degradation, leading to electrode failure after 5 days of continuous current flow. We show that graphene, a commonly used passivation layer for these electrodes, slows nanowire degradation and creates a more uniform surface temperature under current flow. However, the graphene does not prevent Joule heating in the nanowires and local points of high temperature ultimately shift the failure mechanism from nanowire degradation to melting of the underlying plastic substrate. In this paper, surface temperature mapping, lifetime testing under current flow, post-mortem analysis, and modelling illuminate the behaviour and failure mechanisms of nanowires under extended current flow and provide guidelines for managing Joule heating.

Efficient central nervous system AAVrh10-mediated intrathecal gene transfer in adult and neonate rats.

Intracerebral administration of recombinant adeno-associated vector (AAV) has been performed in several clinical trials. However, delivery into the brain requires multiple injections and is not efficient to target the spinal cord, thus limiting its applications. To assess widespread and less invasive strategies, we tested intravenous (IV) or intrathecal (that is, in the cerebrospinal fluid (CSF)) delivery of a rAAVrh10-egfp vector in adult and neonate rats and studied the effect of the age at injection on neurotropism. IV delivery is more efficient in neonates and targets predominantly Purkinje cells of the cerebellum and sensory neurons of the spinal cord and dorsal root ganglia. A single intra-CSF administration of AAVrh10, single strand or oversized self-complementary, is efficient for the targeting of neurons in the cerebral hemispheres, cerebellum, brainstem and spinal cord. Green fluorescent protein (GFP) expression is more widespread in neonates when compared with adults. More than 50% of motor neurons express GFP in the three segments of the spinal cord in neonates and in the cervical and thoracic regions in adults. Neurons are almost exclusively transduced in neonates, whereas neurons, astrocytes and rare oligodendrocytes are targeted in adults. These results expand the possible routes of delivery of AAVrh10, a serotype that has shown efficacy and safety in clinical trials concerning neurodegenerative diseases.

Body mass index, cardiorespiratory fitness and cardiometabolic risk factors in youth from Portugal and Mozambique.

OBJECTIVES: The objectives of this study are to examine differences in cardiometabolic risk indicators, as well as their prevalences, in Portuguese and Mozambican youth, and to investigate the associations between weight status and cardiorespiratory fitness levels with cardiometabolic risk. METHODS: The sample comprises 721 adolescents (323 Mozambican and 398 Portuguese), aged 10-15 years. Anthropometry (height, sitting height, weight and waist circumference), blood pressure, serum-fasting triglycerides, high-density lipoprotein cholesterol and glucose, and cardiorespiratory fitness were measured. Maturity offset was estimated and a cardiometabolic risk score adjusted for sex, age and biological maturity was computed. Adolescents were classified as normal weight and overweight/obese as well as fit or unfit (cardiorespiratory fitness). RESULTS: Portuguese youth have better cardiometabolic and cardiorespiratory fitness profiles. About 32% and 30% of Portuguese boys and girls, respectively, are overweight/obese; in Mozambicans, these prevalences are 7.5% for boys and 21% for girls; in addition, 81.6% of Portuguese boys and 77.7% of Portuguese girls were classified as cardiorespiratory fit, against 54% and 44.4% of Mozambican boys and girls, respectively. No statistically significant differences (P>0.05) were found between Mozambicans and Portuguese for the cluster of three or more cardiometabolic risk indicators. A positive relationship (P<0.001) was found between weight status and cardiometabolic risk in adolescents from both countries; however, a negative association (P<0.001) between cardiorespiratory fitness and cardiometabolic risk was only found among Portuguese youth. CONCLUSIONS: Portuguese and Mozambican youth differ in their cardiometabolic risk profiles, body weight and cardiorespiratory fitness, favoring Portuguese. Overweight/obesity and low cardiorespiratory fitness levels are related to a worse cardiometabolic risk profile, being relevant to design public health intervention strategies to reduce excess weight and increase cardiorespiratory fitness.

Sensitivity of field isolates of Eimeria acervulina and E. maxima from three regions in Brazil to eight anticoccidial drugs.

Rotation with different active ingredients is among the most effective and recommended strategies to preserve the efficacy of anticoccidial drugs and reduce the emergence of resistance. Tools such as anticoccidial sensitivity tests (ASTs) are ideally used to make rational rotation programs and bring benefits to production. The objective of this study was to evaluate the sensitivity of E. acervulina (EA) and E. maxima (EM) from 3 different regions in Brazil, by using four ASTs. Feces samples weighing 6 to 7 kg were collected in the regions of São Paulo, Paraná, and Minas Gerais. Prevalent oocysts from feces were filtered, identified, and quantified to conduct 2 ASTs with EA and 2 with EM. The same experimental design was used in every AST (4 replicates per treatment, with 6 birds each, for a total of 240 birds). Treatment groups were a nonchallenged and nonmedicated control group (T1), a challenged and nonmedicated control group (T2), and the other groups challenged and treated with the following compounds: lasalocid (90 ppm - T3), maduramycin (6 ppm - T4), decoquinate (30 ppm - T5), nicarbazin+semduramicin (66 ppm - T6), monensin (110 ppm - T7), salinomycin (66 ppm - T8), narasin+nicarbazin (100 ppm - T9), and nicarbazin (125 ppm - T10). At the end of each AST (20 d), the percent change (delta value) between the treated group (T3 to T10) and the control group (T2) was calculated for the following variables: body weight gain, feed conversion ratio, lesion score, and an indicator of percentage of optimal anticoccidial activity (POAA) that included T2. Different sensitivity levels of EA and EM isolates could be identified. As a whole, drugs from T5 and T3 groups showed higher delta values when compared to other compounds, whereas the lowest sensitivity levels of these isolates were observed in groups T4 and T7. Despite some limiting factors, ASTs can be a good tool for strategic selection of anticoccidial drugs in order to maintain efficacy and extend the lifespan of these molecules.

Survivin drives tumor-associated macrophage reprogramming: a novel mechanism with potential impact for obesity.

PURPOSE: Recent studies point to adipose-derived stem cells (ASCs) as a link between obesity and cancer. We aimed to determine whether survivin, which is highly secreted by ASCs from subjects with obesity, might drive a pro-tumoral phenotype in macrophages. METHODS: The effect of ASC conditioned medium on the macrophage phenotype was assessed by expression studies. Survivin intracellular localization and internalization were examined by subcellular fractionation and immunofluorescence, respectively. Loss- and gain-of-function studies were performed using adenoviral vectors, and gene expression patterns, migration and invasion capacities of cancer cells were examined. Heterotypic cultures of ASCs, macrophages and cancer cells were established to mimic the tumor microenvironment. Survivin-blocking experiments were used to determine the impact of survivin on both macrophages and cancer cells. Immunohistochemical analysis of survivin was performed in macrophages from ascitic fluids of cancer patients and healthy controls. RESULTS: We found that obese-derived ASCs induced a phenotypic switch in macrophages characterized by the expression of both pro- and anti-inflammatory markers. Macrophages were found to internalize extracellular survivin, generating hybrid macrophages with a tumor-associated phenotype that included secretion of survivin. Exogenous expression of survivin in macrophages generated a similar phenotype and enhanced the malignant characteristics of cancer cells by a mechanism dependent on survivin phosphorylation at threonine 34. Survivin secreted by both ASCs from subjects with obesity and tumor-associated macrophages synergistically boosted the malignancy of cancer cells. Importantly, survivin was mainly detected in ascites-associated macrophages from patients with a malignant diagnosis. CONCLUSION: Our data indicate that survivin may serve as a molecular link between obesity and cancer and as a novel marker for tumor-associated macrophages.

Towards diversity in science - a glance at gender disparity in the Brazilian Society of Neuroscience and Behavior (SBNeC).

Gender equity is far from being achieved in most academic institutions worldwide. Women representation in scientific leadership faces multiple obstacles. Implicit bias and stereotype threat are considered important driving forces concerning gender disparities. Negative cultural stereotypes of weak scientific performance, unrelated to true capacity, are implicitly associated with women and other social groups, influencing, without awareness, attitudes and judgments towards them. Meetings of scientific societies are the forum in which members from all stages of scientific careers are brought together. Visibility in the scientific community stems partly from presenting research as a speaker. Here, we investigated gender disparities in the Brazilian Society of Neuroscience and Behavior (SBNeC). Across the 15 mandates (1978-2020), women occupied 30% of the directory board posts, and only twice was a woman president. We evaluated six meetings held between 2010 and 2019. During this period, the membership of women outnumbered that of men in all categories. A total of 57.50% of faculty members, representing the potential pool of speakers and chairs, were female. Compared to this expected value, female speakers across the six meetings were scarce in full conferences (χ2(5)=173.54, P<0.001) and low in symposia (χ2(5)=36.92, P<0.001). Additionally, women chaired fewer symposia (χ2(5)=47.83, P<0.001). Furthermore, men-chaired symposia had significantly fewer women speakers than women-chaired symposia (χ2(1)=56.44, P<0.001). The gender disparities observed here are similar to those in other scientific societies worldwide, urging them to lead actions to pursue gender balance and diversity. Diversity leads not only to fairness but also to higher-quality science.

SET protein (TAF1beta, I2PP2A) is involved in neuronal apoptosis induced by an amyloid precursor protein cytoplasmic subdomain.

When overexpressed, a short cytoplasmic domain of the amyloid precursor protein (APP), normally unmasked in the brain of Alzheimer's disease patients, activates caspase-3 and induces neuronal death. Death induction by this "Jcasp" domain is lost when tyrosine 653 is changed into an aspartate, suggesting specific interactions with unknown partners. To identify these putative partners and start to elucidate the mechanisms involved in Jcasp-induced cell death, we internalized a biotinylated version of the peptide into primary neurons and analyzed intracellular interacting proteins by pull-down and mass spectrometry. We find that SET protein, also called template-activating factor (TAF1beta) or phosphatase 2A inhibitor 2 (I2(PP2A)), specifically binds Jcasp early after internalization and that SET and Jcasp interact directly in vitro. Down-regulation of SET reduces Jcasp-induced cell death, confirming a role of this protein in Jcasp-induced apoptosis. Conversely, SET gain of function increases cell death, which suggests that SET level is crucial for neuronal survival/death. Taken together, these results suggest that SET is part of a neuronal apoptotic pathway related to Alzheimer's disease and provides a new entry in the analysis of this pathology.

Towards diversity in science - a glance at gender disparity in the Brazilian Society of Neuroscience and Behavior (SBNeC)

Gender equity is far from being achieved in most academic institutions worldwide. Women representation in scientific leadership faces multiple obstacles. Implicit bias and stereotype threat are considered important driving forces concerning gender disparities. Negative cultural stereotypes of weak scientific performance, unrelated to true capacity, are implicitly associated with women and other social groups, influencing, without awareness, attitudes and judgments towards them. Meetings of scientific societies are the forum in which members from all stages of scientific careers are brought together. Visibility in the scientific community stems partly from presenting research as a speaker. Here, we investigated gender disparities in the Brazilian Society of Neuroscience and Behavior (SBNeC). Across the 15 mandates (1978-2020), women occupied 30% of the directory board posts, and only twice was a woman president. We evaluated six meetings held between 2010 and 2019. During this period, the membership of women outnumbered that of men in all categories. A total of 57.50% of faculty members, representing the potential pool of speakers and chairs, were female. Compared to this expected value, female speakers across the six meetings were scarce in full conferences (χ2(5)=173.54, P<0.001) and low in symposia (χ2(5)=36.92, P<0.001). Additionally, women chaired fewer symposia (χ2(5)=47.83, P<0.001). Furthermore, men-chaired symposia had significantly fewer women speakers than women-chaired symposia (χ2(1)=56.44, P<0.001). The gender disparities observed here are similar to those in other scientific societies worldwide, urging them to lead actions to pursue gender balance and diversity. Diversity leads not only to fairness but also to higher-quality science.

Antinociceptive properties of the methanolic extract obtained from Ipomoea pes-caprae (L.) R. Br.

Ipomoea pes-caprae is a medicinal plant used in many countries for the treatment of several ailments, including inflammatory and algesic processes. The present study describes the antinociceptive effects of the methanolic extract and two fractions obtained from aerial parts of this plant. The results indicated that both methanolic extract and two fractions (ethyl acetate and aqueous) exhibited considerable antinociceptive activity against two classical models of pain in mice. Methanolic extract presented a calculated ID50 value of 33.8 mg/kg, i.p. against writhing test and also inhibited both phases of pain (neurogenic and inflammatory) of the formalin test with ID50 of 37.7 and 12.5 mg/kg, i.p. for the first and second phase, respectively. Preliminary phytochemical analysis suggested the presence of steroids, terpenoids, alkaloids and flavonoids. These findings support, at least in part, the popular use of I. pes-caprae to treat dolorous processes.

Isolation and identification of compounds with antinociceptive action from Ipomoea pes-caprae (L.) R. Br.

This study describes the isolation and identification of several constituents from Ipomoea pes-caprae (L.) R. Br., a medicinal plant frequently employed in folk medicine of many countries as a remedy against several diseases, including inflammation and pain. Our results demonstrate that some of these compounds, such as glochidone, betulinic acid, alpha- and beta-amyrin acetate, isoquercitrin, etc. showed pronounced antinociceptive properties in the writhing test and formalin test in mice. These data confirm our previous work concerning the antinociceptive action of the hydroalcoholic extract of I. pes-caprae and justify, at least in part, the popular use of this plant for the treatment of dolorous processes.

[Medical audit: prenatal care program in a health center in Southern Brazil]. / Auditoria médica: programa de pré-natal em posto de saúde na região Sul do Brasil.

INTRODUCTION: A medical audit on the prenatal care program in the Vila Municipal Health Center, Pelotas, RS, Brazil, was described with the purpose of verifying the aspects of the medical process and improving the program's effectiveness. METHODS: Data from prenatal specific records were collected. Pregnant women with delivery due date in 1997 and in the first semester of 1998 were included in the study. Women registered in the program when they were 4-month pregnant and who had had at least 5 visits were also enrolled. Bivariate analysis was used to detect health care indicators. RESULTS: A total of 73 pregnant women were registered in the program in 1997 and 75 in 1998. In 1997, the average number of medical visits was 5.2, while in 1998 this average was 6.2. The difference between the means was statistically significant (p<0.05). Some medical process indicators were analysed to verify the quality of the care. CONCLUSION: The use of the epidemiological method to organize health services was discussed. This type of study requires few resources and time and it can provide guidelines to health service actions.

[The refusal of nursing consultation. A comprehensive analysis of the phenomenon]. / O abandono da consulta de enfermagem uma análise compreensiva do fenômeno.

The purpose of this study was to understand the abandonment of nursing consultation focused on children 0 to 2 years old. A phenomenological approach was used to make apprehension of the essence of the phenomenon and to try to understand it without becoming attached to its determinant causes. Interviews were done with 12 mothers to describe their experiences on nursing consultation and, therefore, making possible to see the essence of the phenomenon.

[Adolescent mothers caring for their child]. / Mãe adolescente cuidando do filho.

This is a qualitative research that had the objective of understanding the perception of adolescent mothers about their child[symbol: see text]s care. The phenomenological approach was used to analyze data gathered from 7 mothers. To the adolescent mothers caring for their child means feeding the child, do his/her hygiene, care for his/her illness, promote rest, give attention, give love, give educational support and promote leisure. These themes converge to 3 main categories: attend to the biological, psychological and social necessities of the child.

[An analysis of the nursing care given to the child on parenteral nutrition]. / Análise da assistência de enfermagem prestada à criança em nutrição parenteral.

The purpose of this study was to collect information on nursing car of children receiving parenteral nutrition in order to gain knowledge to improve nursing care according to the resources available in the hospital. Our results showed that in the three hospitals studied there were no adequate physical space to prepare parenteral nutrition. There were no suitable accommodations for the children, the nursing care on P.N. were not standardized and the follow up was not well done.

[Evaluation of the nursing care for children 0-2 years old in a health center]. / Avaliação da consulta de enfermagem prestada à criança de 0 a 2 anos de idade em um centro de saúde.

The study is an exploratory-descriptive research that has as a main goal to evaluate the nursing consultation to 0 to 2 years old children in a health center. To do that the authors analysed the data collected in the children records from 1985 to 1988 and from interviews with mothers whose children had the nursing consultation in that time.

Molecular characteristics of Human Endogenous Retrovirus type-W in schizophrenia and bipolar disorder.

Epidemiological and genome-wide association studies of severe psychiatric disorders such as schizophrenia (SZ) and bipolar disorder (BD), suggest complex interactions between multiple genetic elements and environmental factors. The involvement of genetic elements such as Human Endogenous Retroviruses type 'W' family (HERV-W) has consistently been associated with SZ. HERV-W envelope gene (env) is activated by environmental factors and encodes a protein displaying inflammation and neurotoxicity. The present study addressed the molecular characteristics of HERV-W env in SZ and BD. Hundred and thirty-six patients, 91 with BD, 45 with SZ and 73 healthy controls (HC) were included. HERV-W env transcription was found to be elevated in BD (P<10-4) and in SZ (P=0.012) as compared with HC, but with higher values in BD than in SZ group (P<0.01). The corresponding DNA copy number was paradoxically lower in the genome of patients with BD (P=0.0016) or SZ (P<0.0003) than in HC. Differences in nucleotide sequence of HERV-W env were found between patients with SZ and BD as compared with HC, as well as between SZ and BD. The molecular characteristics of HERV-W env also differ from what was observed in Multiple Sclerosis (MS) and may represent distinct features of the genome of patients with BD and SZ. The seroprevalence for Toxoplasma gondii yielded low but significant association with HERV-W transcriptional level in a subgroup of BD and SZ, suggesting a potential role in particular patients. A global hypothesis of mechanisms inducing such major psychoses is discussed, placing HERV-W at the crossroads between environmental, genetic and immunological factors. Thus, particular infections would act as activators of HERV-W elements in earliest life, resulting in the production of an HERV-W envelope protein, which then stimulates pro-inflammatory and neurotoxic cascades. This hypothesis needs to be further explored as it may yield major changes in our understanding and treatment of severe psychotic disorders.