Characterization of the nigroestriatal system in a sample of patients with amyotrophic lateral sclerosis / Caracterização do sistema nigroestriatal em uma amostra de pacientes com esclerose lateral amiotrófica

Abstract Background The coexistence of amyotrophic lateral sclerosis (ALS) with clinical forms of Parkinson disease (PD), although uncommon, is found to a greater degree than one would expect by chance. The pathological mechanisms of ALS and PD are still not fully understood, and the coexistence of these two diseases suggests that they could share mechanisms in common. Objective Here we present a sample of patients with clinically definitive or probable ALS who were evaluated with single-photon emission computed tomography SPECT/TRODAT and compared with non-ALS controls. Methods Patients with clinically definite or probable ALS were assessed with the amyotrophic lateral sclerosis functional rating scale (ALSFRS) to define severity and had their demographic data collected. The TRODAT results of patients with ALS were compared with those of patients with a diagnosis of PD with less than 10 years of duration, and with patients with a diagnosis of others movement disorders not associated with neurodegenerative diseases. Results A total of 75% of patients with ALS had TRODAT results below the levels considered normal; that was also true for 25% of the patients in the control group without neurodegenerative disease, and for 100% of the patients in the PD group. A statistically significant difference was found between patients with ALS and the control group without neurodegenerative disease in the TRODAT values < 0.05. Conclusions Our study fits with the neuropathological and functional evidence that demonstrates the existence of nigrostriatal dysfunction in patients with ALS. Further research to better understand the role of these changes in the pathophysiological process of ALS needs to be performed.

Resumo Antecedentes A coexistência da esclerose lateral amiotrófica (ELA) com formas clínicas da doença de Parkinson (DP), embora incomum, é encontrada em um grau maior do que seria esperado ao acaso. Os mecanismos patológicos da ELA e da DP ainda não são totalmente compreendidos e a coexistência dessas duas doenças sugere que elas podem compartilhar mecanismos em comum. Objetivo Apresentamos uma amostra de pacientes com ELA clinicamente definida ou provável que foram avaliados com tomografia computadorizada por emissão de fóton único (SPECT)/TRODAT e comparados com controles sem ELA. Métodos Pacientes com ELA clinicamente definida ou provável foram avaliados com a escala funcional de esclerose lateral amiotrófica (ALSFRS) para definir a gravidade e foram coletados os seus dados demográficos. Os resultados do TRODAT de pacientes com ELA foram comparados com aqueles de pacientes com diagnóstico de DP com menos de 10 anos de duração e com pacientes com diagnóstico de outros distúrbios do movimento não associados a doenças neurodegenerativas. Resultados Um total de 75% dos pacientes com ELA apresentou resultados de TRODAT abaixo dos níveis considerados normais; 25% no grupo controle sem doença neurodegenerativa e 100% no grupo DP. Uma diferença estatisticamente significativa foi encontrada entre os pacientes com ELA e o grupo controle sem doença neurodegenerativa nos valores de TRODAT p< 0,05. Conclusões Nosso estudo está de acordo com as evidências neuropatológicas e funcionais que demonstram a existência de disfunção nigroestriatal em pacientes com ELA. Mais pesquisas para entender melhor o papel dessas mudanças no processo fisiopatológico da ELA precisam ser realizadas.

Ivermectina: ¿Un antiparasitario frente a SARS-CoV-2? / Ivermectin: an antiparasitic drug to fight SARS-CoV-2?

RESUMEN La capacidad de propagación y letalidad del SARS-CoV-2 en todo el mundo motiva la urgente necesidad de desarrollar una estrategia terapéutica apropiada para controlar los casos de COVID-19. El desarrollo de nuevos fármacos frente a este nuevo virus es apremiante debido a su rápida diseminación. Se han propuesto alternativas paralelas empleando fármacos ya disponibles para fines similares. Esta revisión describe el potencial antiviral de la ivermectina, así como sus mecanismos de acción frente a algunos virus, y discute su probable aplicación contra el SARS-CoV-2.

ABSTRACT The global spread and lethality of SARS-CoV-2 prompt the urgent need to develop an appropriate therapeutic strategy to control COVID-19 cases. The development of new drugs to fight this novel virus is urgent due to its rapid spread. Parallel alternatives have been proposed by using drugs already available for similar purposes. This review article describes the antiviral potential of ivermectin as well as its mechanisms of action against some viruses, and discusses its probable use to fight SARS-CoV-2.

Verapamil inhibits efflux pumps in Candida albicans, exhibits synergism with fluconazole, and increases survival of Galleria mellonella.

The emergence of resistance requires alternative methods to treat Candida albicans infections. We evaluated efficacy of the efflux pump inhibitor (EPI) verapamil (VER) with fluconazole (FLC) against FLC-resistant (CaR) and -susceptible C. albicans (CaS). The susceptibility of both strains to VER and FLC was determined, as well as the synergism of VER with FLC. Experiments were performed in vitro for planktonic cultures and biofilms and in vivo using Galleria mellonella. Larval survival and fungal recovery were evaluated after treatment with VER and FLC. Data were analyzed by analysis of variance and Kaplan-Meier tests. The combination of VER with FLC at sub-lethal concentrations reduced fungal growth. VER inhibited the efflux of rhodamine 123 and showed synergism with FLC against CaR. For biofilms, FLC and VER alone reduced fungal viability. The combination of VER with FLC at sub-lethal concentrations also reduced biofilm viability. In the in vivo assays, VER and FLC used alone or in combination increased the survival of larvae infected with CaR. Reduction of fungal recovery was observed only for larvae infected with CaR and treated with VER with FLC. VER reverted the FLC-resistance of C. albicans. Based on the results obtained, VER reverted the FLC-resistance of C. albicans and showed synergism with FLC against CaR. VER also increased the survival of G. mellonella infected with CaR and reduced the fungal recovery.

Beta-adrenergic pathway activation in head and neck cancer: a comprehensive analysis of the clinical and genomic features using integrated bioinformatics / Ativação da via beta-adrenérgica no câncer de cabeça e pescoço: uma análise abrangente das características clínicas e gênomicas utilizando a bioinformática

O estresse crônico leva à ativação da via de sinalização beta-adrenérgica. Sua ativação tem sido implicada na progressão de diferentes tipos de câncer, mas seu papel nos carcinomas espinocelulares de cabeça e pescoço (CECPs) permanece indefinido. O objetivo deste estudo foi investigar o papel da ativação da via betaadrenérgica na progressão dos CECPs, avaliar seu impacto na sobrevida dos pacientes e buscar possíveis terapias para pacientes que encontravam-se com a via beta-adrenérgica ativa. Quinhentos e vinte pacientes do The Cancer Genome Atlas com CECPs primários foram divididos em dois grupos: ADRB2baixa / SLC6A2baixa e ADRB2alta / SLC6A2alta. A associação de características clinicopatológicas e genômicas entre os grupos foram analisadas utilizando bioinformática. Os genes diferencialmente expressos (DEGs) foram identificados através da análise da expressão diferencial. A análise de sobrevida também foi realizada com base nas expressões ADRB2 e SLC6A2. Foram identificados medicamentos em potencial para tratamento de CECPs com base nos DEGs. Houve associação entre as expressões ADRB2 e SLC6A2 com idade, raça, localização do tumor, grau histológico, invasão perineural e status do HPV p16. Foram identificados 898 DEGs entre os grupos. Foi demonstrado que a expressão ADRB2alta / SLC6A2alta influenciou a proliferação, adesão e invasão de células CECPs além da angiogênese. Pacientes com carcinomas espinocelular de laringe e faringe apresentando expressão ADRB2alta / SLC6A2alta tiveram menor sobrevida. Por fim, 56 drogas antineoplásicas e imunoterápicas aprovadas pelo Food Drugs Administration foram identificadas como potenciais alvos para o tratamento personalizado. Significância: Estes achados sugerem fortemente um papel proeminente da sinalização beta-adrenérgica no CECPs ao estimular um fenótipo tumoral mais agressivo. Estas alterações tiveram um impacto negativo no prognóstico dos pacientes com CECP em região de faringe e laringe(AU)

Chronic stress leads to the activation of the beta-adrenergic pathway. Its activation has been implicated in the progression of different types of cancer but its role on head and neck squamous cell carcinomas (HNSCCs) remains undefined. The aim of this study was to investigate the influence of the beta-adrenergic pathway activation in the progression of HNSCCs, assess its impact in the survival of the patients, and explore the potential targets. Five hundred and twenty The Cancer Genome Altas patients with primary HNSCCs were divided in two groups: ADRB2low / SLC6A2low and ADRB2high / SLC6A2high. The association of clinicopathological and genomic features between the groups was analyzed using a bioinformatic approach. Differentially expressed genes (DEGs) were identified through differential expression analysis. Survival analysis was also performed based on ADRB2 and SLC6A2 expressions. Potential drugs for treatment of HNSCC were identified based on the DEGs. There was association between ADRB2 and SLC6A2 expressions with age, race, tumor site, histologic grade, perineural invasion, and HPV p16 status. It was identified 898 DEGs between the groups. It was demonstrated that ADRB2high / SLC6A2high expression influenced HNSCC cells proliferation, adhesion, invasion, and angiogenesis. Patients with larynx and pharynx squamous cell carcinomas presenting ADRB2high / SLC6A2high expression showed had lower survival rates. Finally, 56 Food Drugs Administration-approved antineoplastic and immunotherapeutic drugs were identified as potential targets for the personalized treatment. Significance: These findings strongly suggest a prominent role of beta-adrenergic pathway in HNSCC by stimulating a more aggressive tumoral phenotype. These alterations were shown to negatively impact the prognosis of patients with larynx and pharynx squamous cell carcinomas(AU)

Immunohistochemical expressionof sodium-dependent glucose transporter - 2 (SGLT-2) in clear cell renal carcinoma: possible prognostic implications

ABSTRACT Purpose: Glucose is a major energy resource for tumor cell survival and growth, and its influx into cells is mainly carried out by facilitative glucose transporters (GLUTs). Sodium - dependent glucose transporters (SGLTs) have been highlighted as playing important roles in diabetic treatment. However, their potential roles in cancer remain unclear. We examined expression patterns of SGLTs in tumor tissues together with conventional pathological variables to determine prognostic significance in patients with renal cell carcinoma (RCC). Materials and Methods: Nephrectomy specimens were obtained from 68 patients. GLUT - 1, - 2 and SGLT - 1, - 2 expression in tumor and adjacent normal tissues were analyzed by immunohistochemical staining, and intensity was quantified using an image analyzer. Results: The four glucose transporters evaluated were broadly distributed in tumor tissues as well as throughout the normal parenchyma. There was no significant correlation between transporter expression and conventional pathological variables. However, increased SGLT - 2 expression was significantly associated with shorter overall survival (p < 0.01), regardless of metastatic status. Conclusions: We propose possible prognostic significance of SGLT - 2 expression in human RCC. Given that glucose is a major energy resource for tumor cells and that glucose transport is largely mediated by SGLT, SGLT - 2 may serve as a possible therapeutic target in RCC.

Immunohistochemical expressionof sodium-dependent glucose transporter - 2 (SGLT-2) in clear cell renal carcinoma: possible prognostic implications.

PURPOSE: Glucose is a major energy resource for tumor cell survival and growth, and its influx into cells is mainly carried out by facilitative glucose transporters (GLUTs). Sodium - dependent glucose transporters (SGLTs) have been highlighted as playing important roles in diabetic treatment. However, their potential roles in cancer remain unclear. We examined expression patterns of SGLTs in tumor tissues together with conventional pathological variables to determine prognostic significance in patients with renal cell carcinoma (RCC). MATERIALS AND METHODS: Nephrectomy specimens were obtained from 68 patients. GLUT - 1, - 2 and SGLT - 1, - 2 expression in tumor and adjacent normal tissues were analyzed by immunohistochemical staining, and intensity was quantified using an image analyzer. RESULTS: The four glucose transporters evaluated were broadly distributed in tumor tissues as well as throughout the normal parenchyma. There was no significant correlation between transporter expression and conventional pathological variables. However, increased SGLT - 2 expression was significantly associated with shorter overall survival (p < 0.01), regardless of metastatic status. CONCLUSIONS: We propose possible prognostic significance of SGLT - 2 expression in human RCC. Given that glucose is a major energy resource for tumor cells and that glucose transport is largely mediated by SGLT, SGLT - 2 may serve as a possible therapeutic target in RCC.

Reduced Hepatocellular Expression of Canalicular Transport Proteins in Infants with Neonatal Cholestasis and Congenital Hypopituitarism.

OBJECTIVE: To assess whether prolonged neonatal cholestasis, described in congenital hypopituitarism and septo-optic dysplasia (SOD), is associated with altered expression of selected canalicular ectoenzymes and canalicular transport proteins. STUDY DESIGN: Children with congenital hypopituitarism (n = 21), SOD (n = 18), and cholestasis seen in our center over 26 years were reviewed. Histopathologic findings in archival liver biopsy specimens were assessed (n = 10) and in those with low/normal levels of serum γ-glutamyltransferase (GGT) activity despite conjugated hyperbilirubinemia, expression of canalicular ectoenzymes and canalicular transport proteins was evaluated immunohistochemically. RESULTS: Patients presented at a median age of 8 weeks (range 3-20 weeks) with median total bilirubin 116 µmol/L (45-287 µmol/L), GGT 95 IU/L (25-707 UI/L), and serum cortisol 51 nmol/L (17-240 nmol/L). All but 3 had low free thyroxin (median 9.6 pmol/L [6.8-26.9]) with increased thyroid-stimulating hormone levels (median 5.95 mU/L [<0.1-9.24]). Liver histologic features included moderate-to-severe intralobular cholestasis with nonspecific hepatitis, giant-cell transformation of hepatocytes, and fibrosis. In all, immunohistochemical staining for canalicular ectoenzymes and canalicular transport proteins revealed a degree of reduced expression, associated with normal serum GGT values in 6 of the 10 patients, and another 6 nonbiopsied infants with cholestasis also had low/normal serum GGT activity. Sequencing of ABCB11 and ATP8B1 performed in 6 of the biopsied patients did not identify pathogenic mutations. Following replacement therapy, biochemical evidence of hepatobiliary injury resolved in all children within a median period of 6 months. CONCLUSION: Hepatobiliary involvement in congenital hypopituitarism associated with SOD has a good prognosis, but its etiology remains uncertain. Immunohistochemical expression of canalicular transport proteins was reduced in available liver samples.

Horizontal Gene Transfer Building Prokaryote Genomes: Genes Related to Exchange Between Cell and Environment are Frequently Transferred.

Horizontal gene transfer (HGT) has a major impact on the evolution of prokaryotic genomes, as it allows genes evolved in different contexts to be combined in a single genome, greatly enhancing the ways evolving organisms can explore the gene content space and adapt to the environment. A systematic analysis of HGT in a large number of genomes is of key importance in understanding the impact of HGT in the evolution of prokaryotes. We developed a method for the detection of genes that potentially originated by HGT based on the comparison of BLAST scores between homologous genes to 16S rRNA-based phylogenetic distances between the involved organisms. The approach was applied to 697 prokaryote genomes and estimated that in average approximately 15% of the genes in prokaryote genomes originated by HGT, with a clear correlation between the proportion of predicted HGT genes and the size of the genome. The methodology was strongly supported by evolutionary relationships, as tested by the direct phylogenetic reconstruction of many of the HGT candidates. Studies performed with Escherichia coli W3110 genome clearly show that HGT proteins have fewer interactions when compared to those predicted as vertical inherited, an indication that the number of protein partners imposes limitations to horizontal transfer. A detailed functional classification confirms that genes related to protein translation are vertically inherited, whereas interestingly, transport and binding proteins are strongly enriched among HGT genes. Because these genes are related to the cell exchange with their environment, their transfer most likely contributed to successful adaptation throughout evolution.

Binding targets of termiticidal lectins from the bark and leaf of Myracrodruon urundeuva in the gut of Nasutitermes corniger workers.

BACKGROUND: Lectins, carbohydrate-binding proteins, from the bark (MuBL) and leaf (MuLL) of Myracrodruon urundeuva are termiticidal agents against Nasutitermes corniger workers and have been shown to induce oxidative stress and cell death in the midgut of these insects. In this study, we investigated the binding targets of MuBL and MuLL in the gut of N. corniger workers by determining the effects of these lectins on the activity of digestive enzymes. In addition, we used mass spectrometry to identify peptides from gut proteins that adsorbed to MuBL-Sepharose and MuLL-Sepharose columns. RESULTS: Exoglucanase activity was neutralized in the presence of MuBL and stimulated by MuLL. α-l-Arabinofuranosidase activity was not affected by MuBL but was inhibited by MuLL. Both lectins stimulated α-amylase activity and inhibited protease and trypsin-like activities. Peptides with homology to apolipophorin, trypsin-like enzyme, and ABC transporter substrate-binding protein were detected from proteins that adsorbed to MuBL-Sepharose, while peptides from proteins that bound to MuLL-Sepharose shared homology with apolipophorin. CONCLUSION: This study revealed that digestive enzymes and transport proteins found in worker guts can be recognized by MuBL and MuLL. Thus, the mechanism of their termiticidal activity may involve changes in the digestion and absorption of nutrients. © 2018 Society of Chemical Industry.

Discovery of Novel Leptospirosis Vaccine Candidates Using Reverse and Structural Vaccinology.

Leptospira spp. are diderm (two membranes) bacteria that infect mammals causing leptospirosis, a public health problem with global implications. Thousands of people die every year due to leptospirosis, especially in developing countries with tropical climates. Prophylaxis is difficult due to multiple factors, including the large number of asymptomatic hosts that transmit the bacteria, poor sanitation, increasing numbers of slum dwellers, and the lack of an effective vaccine. Several leptospiral recombinant antigens were evaluated as a replacement for the inactivated (bacterin) vaccine; however, success has been limited. A prospective vaccine candidate is likely to be a surface-related protein that can stimulate the host immune response to clear leptospires from blood and organs. In this study, a comprehensive bioinformatics approach based on reverse and structural vaccinology was applied toward the discovery of novel leptospiral vaccine candidates. The Leptospira interrogans serovar Copenhageni strain L1-130 genome was mined in silico for the enhanced identification of conserved ß-barrel (ßb) transmembrane proteins and outer membrane (OM) lipoproteins. Orthologs of the prospective vaccine candidates were screened in the genomes of 20 additional Leptospira spp. Three-dimensional structural models, with a high degree of confidence, were created for each of the surface-exposed proteins. Major histocompatibility complex II (MHC-II) epitopes were identified, and their locations were mapped on the structural models. A total of 18 ßb transmembrane proteins and 8 OM lipoproteins were identified. These proteins were conserved among the pathogenic Leptospira spp. and were predicted to have epitopes for several variants of MHC-II receptors. A structural and functional analysis of the sequence of these surface proteins demonstrated that most ßb transmembrane proteins seem to be TonB-dependent receptors associated with transportation. Other proteins identified included, e.g., TolC efflux pump proteins, a BamA-like OM component of the ßb transmembrane protein assembly machinery, and the LptD-like LPS assembly protein. The structural mapping of the immunodominant epitopes identified the location of conserved, surface-exposed, immunogenic regions for each vaccine candidate. The proteins identified in this study are currently being evaluated for experimental evidence for their involvement in virulence, disease pathogenesis, and physiology, in addition to vaccine development.

Polimorfismo Ala54Thr do gene fatty acid-binding protein-2 (FABP-2): uma revisão bibliográfica / The Ala54Thr polymorphism of the fatty acid-binding protein-2 (FABP-2) gene: a literature review

O polimorfismo p.Ala54Thr (rs1799883) do gene fatty acid-binding protein-2 (FABP2) tem associação com resistência insulínica, síndrome metabólica e obesidade. A hipótese é de que o alelo mutante aumente a absorção de ácidos graxos intestinais, a concentração lipídica plasmática e a oxidação de gordura. Assim, o objetivo deste trabalho foi revisar o papel do polimorfismo p.Ala54Thr do gene FABP-2 na obesidade. A busca da literatura foi realizada na base de dados MEDLINE, através do PubMed e no Portal de Periódicos de Aperfeiçoamento de Pessoal de Nível Superior (Capes) com termos relacionados com o polimorfismo e obesidade. Parece não haver uma associação significativa da presença do alelo Thr54 com obesidade, apesar de ser uma doença complexa e que possivelmente não tenha sido captada por estudos de associação; diferente do colesterol total e lipoproteína de baixa densidade (low density level cholesterol, LDL-c), maior nos portadores do alelo Thr54. Alterações de adipocitocinas devem estar associadas a estas diferenças de perfil lipídico.

The p.Ala54Thr polymorphism (rs1799883) of the fatty acid-binding Protein-2 (FABP-2) gene has been associated with insulin resistance, metabolic syndrome and obesity. The hypothesis that the mutant allele increases the absorption of fatty acid by the bowel, plasma lipid concentration, and fat oxidation. Thus, the aim of this study was to review the role of FABP-2 Ala54Thr polymorphism in obesity. A literature search was conducted in MEDLINE database, using PubMed and Capes Portal with terms related to polymorphism and obesity. It does not seem to be a significant association between Thr54 allele and obesity, although being a complex disease and that possibly has not been captured by association studies; unlike total cholesterol and low density level cholesterol (LDL-c), which were higher in Thr54 allele carriers. Adipocytokines changes should be associated with these differences in lipid profile.

Efeito da redução da função colinérgica na mecânica pulmonar e na histopatologia pulmonar em modelo experimental de inflamação aguda induzida por instilação de LPS em camundongos geneticamente modificados / Cholinergic function reduction effect of pulmonary mechanics and pulmonary histopathology acute inflammation model of experimental induced by LPS in mice genetically modified

A lesão pulmonar aguda (LPA) é caracterizada por inflamação pulmonar de início súbito com recrutamento de polimorfonucleares e liberação de mediadores próinflamatórios. É uma condição grave que evolui com óbito em aproximadamente 40% dos casos. Diversos estudos que elucidaram a fisiopatologia da LPA, o tratamento ainda é insatisfatório. O sistema colinérgico anti-inflamatório foi descrito no pulmão e está relacionado a um reflexo via nervo vago que inibe a liberação de citocinas inflamatórias por efeitos relacionados a ação da acetilcolina em receptores nicotínicos. Nossa hipótese é de que a redução de VAChT, que está relacionada ao déficit na liberação de ACh, module a resposta inflamatória pulmonar em modelo de LPS. Objetivo: 1. Avaliar se a deficiência de VAChT modula a resposta pulmonar em animais geneticamente modificados; 2. Avaliar se a deficiência colinérgica induzida por redução de VAChT está envolvida na resposta pulmonar ao LPS e elucidar alguns mecanismos envolvidos; 3. Avaliar o potencial terapêutico do PNU, um agonista de alfa7nAChR nas alterações funcionais e histopatológicas em modelo de LPA em animais C57Bl6. Metodologia: Foram utilizados camundongos machos geneticamente modificados mutante (VAChT KDHOM) ou selvagem (WT) e C57BL/6. Inicialmente avaliamos a função pulmonar e a histopatologia pulmonar em animais VAChT KDHOM. Após, animais WT e VAChT KDHOM receberam instilação intranasal de LPS ou salina e a resposta inflamatória foi avaliada de 1,5h até 72 horas após. Ainda, foi avaliado a resposta pulmonar em VAChT KDHOM e WT após a instilação de LPS intraperitoneal. Por fim, animais C57BL/6 instilados com LPS intranasal, receberam tratamento prévio ou após com PNU, agonista do receptor nicotínico alfa7. Resultados: Animais mutante apresentaram maior quantidade de células recuperadas no lavado bronco alveolar (LBA) e aumento de citocinas próinflamatórias, aumento de edema peribrônquico e piora da função pulmonar. Ainda,...

Acute lung injury (ALI) is characterized by acute lung inflammation with recruitment of polymorphonuclear and release of proinflammatory mediators. It is a severe condition since leads to death 40% of the cases. Several studies have elucidated the pathophysiology of ALI, however the treatment is still unsatisfactory. The anti-inflammatory cholinergic system was described in the lung and is related to a vagal nerve reflex that inhibits the release of inflammatory cytokines by the action o ACh on nicotinic receptors. Our hypothesis is that the VAChT reduction, which is related to the deficit in the release of ACh, modulates the pulmonary inflammatory response in a model of LPS. Aim: 1. To assess whether VAChT deficiency modulates the pulmonary response in genetically modified animals; 2. Assess whether cholinergic deficiency induced reduction VAChT is involved in pulmonary response to LPS and elucidate some mechanisms involved; 3. To evaluate the therapeutic potential of PNU, an agonist alfa7nAChR, in functional and histological changes in C57BL6 mice with LPA. Methods: Mutant genetically modified male mice (VAChT KDHOM) or wild (WT) and C57BL/6 were used. First, we evaluated lung function and lung histopathology in VAChT KDHOM animals. After, WT animals and VAChT KDHOM received intranasal instillation of LPS or saline and the inflammatory response was assessed 1.5 hours to 72 hours. Moreover, the pulmonary response was evaluated in WT and VAChT KDHOM after instillation of LPS intraperitoneally. Finally, C57BL6 instilled with intranasal LPS received prior or post-treatment with PNU, an alfa7 nicotinic receptor agonist. Results: Mutant animals had higher number of cells recovered in brochoalveolar lavage (BAL) and increased pro-inflammatory cytokines, peribronchial edema and worsening of lung function. Still, there was an increase of NF_kB expression and reduction of JAK2. The VAChT deficiency induced increase in inflammatory cells...

Avaliação da imunoexpressão do transportador de glicose-1 em cistos radiculares e dentígeros

Dentre as lesões que levam à destruição óssea nos maxilares, os cistos odontogênicos são as mais comuns. A inflamação que participa na patogênese de alguns desses cistos pode estimular modificações no metabolismo celular energético, possibilitando a expressão do transportador de glicose-1 (GLUT-1). A partir deste contexto, o presente estudo objetiva analisar a imuno expressão do GLUT-1 em cistos radiculares (CR) e dentígeros (CD).A amostra foi constituída de 36 casos, sendo 18 de CR e 18 de CD. Foi realizado estudo morfológico para o diagnóstico histopatológico de coloração por hematoxilina & eosina (HE),bem como a análise da intensidade do infiltrado inflamatório dos CRs (leve/intenso) no aumento de 100x. Para avaliação imuno-histoquímica, foi utilizada a técnica da estreptoavidina-biotina, com o uso do anticorpo anti-GLUT-1 (GeneTex®, 1:300, citrato pH6, Pascal). A análise quantitativa foi realizada por meio de contagem percentual de células imunomarcadas em cinco campos fotografados no aumento de 400x; enquanto a análise da intensidade de imunomarcação (sem expressão/fraca expressão/forte expressão) ocorreu através da avaliação de um campo fotografado no aumento de 100x; para ambas as avaliações,utilizou-se o programa Image J. Os dados clínicos foram comparados entre grupos por meio do teste qui-quadrado de Pearson ou Exato de Fisher e por meio de Regressão Logística Multinomial. As contagens de células positivas foram analisadas por meio do teste de Mann Whineyou Kruskall-Wallis/Dunn (dados não paramétricos). Adicionalmente, utilizou-se a correlação de Spearman entre alguns grupos. Todos os testes estatísticos tiveram como baseos níveis de significância de 5%...

Odontogenic cysts are one of the most common osseous-destructive lesions in the jaws.Inflammation involved in the pathogenesis of some of these cysts can stimulate changes in theenergy cellular metabolism, enabling an increase of glucose transporter-1 expression (GLUT1).In this way, the present study aims to evaluate the immunohistochemical expression ofGLUT-1 in radicular cysts (RC) and dentigerous cysts (DC). The sample consists of 36 cases,18 RC and 18 cases of DC. Morphological study was conducted for histopathologic diagnosisby staining with HE as well as the intensity of the RCs’ inflammatory infiltrate analysis(slight/heavy) using 100x magnification. For immunohistochemical assessment, the techniquestreptavidin-biotin was used, with anti-GLUT-1 antibody (GeneTex®, 1: 300, Citrate pH 6).Quantitative analysis was performed by counting of immunostained cells in 05 photographedfields at 400x magnification. The analysis of immunostaining intensity (no expression / weakexpression / strong expression) occurred by evaluating 01 photographed field at 100xmagnification. The software Image J was used for both reviews. Clinical data between groupswere compared using Pearson's chi-square test or Fisher's exact test and through LogisticMultinomial Regression. Scores of positive cells were analyzed using the Mann-Whiney orKruskall-Wallis/Dunn test (nonparametric data). Additionally, the Spearman correlation wasused among some groups. All statistical tests were based on the significance level of 5%...

El ácido nicotínico aumenta el transporte celular de colesterol de las lipoproteínas de alta densidad en pacientes con hipoalfalipoproteinemia / Nicotinic acid increases cellular transport of high density lipoprotein cholesterol in patients with hypoalphalipoproteinemia

Background: Plasma high density lipoproteins (HDL) are involved in reverse cholesterol transport mediated by the scavenger receptor class B type I (SR-BI). Nicotinic acid increases HDL cholesterol levels, even though its specific impact on SR-BI dependent-cellular cholesterol transport remains unknown. Aim: To determine the effect of nicotinic acid on HDL particle functionality in cholesterol efflux and uptake mediated by SR-BI in cultured cells in hypoalphalipoproteinemic patients. Material and Methods: In a pilot study, eight patients with low HDL (≤ 40 mg/dL) were treated with extended release nicotinic acid. HDL cholesterol and phospholipid levels, HDL2 and HDL3 fractions and HDL particle sizes were measured at baseline and post-therapy. Before and after nicotinic acid treatment, HDL particles were used for cholesterol transport studies in cells transfected with SR-BI. Results: Nicotinic acid treatment raised total HDL cholesterol and phospholipids, HDL2 levels as well as HDL particle size. Nicotinic acid significantly increased HDL cholesterol efflux and uptake capacity mediated by SR-BI in cultured cells. Conclusions: Nicotinic acid therapy increases SR-BI-dependent HDL cholesterol transport in cultured cells, establishing a new cellular mechanism by which this lipid-lowering drug appears to modulate HDL metabolism in patients with hypoalphalipoproteinemia.

Impact of a cholesterol membrane transporter's inhibition on vitamin D absorption: A double-blind randomized placebo-controlled study.

Oral supplements are important to prevent and treat vitamin D deficiency. Despite the growing number of prescriptions, vitamin D's absorptive mechanisms are not clearly elucidated. By evaluating the effect of ezetimibe on vitamin D absorption, we aim to determine if the cholesterol transporter Niemann-Pick C1-Like 1 transporter contributes to it. This randomized, double-blind, placebo-controlled trial (ClinicalTrials.govNCT02234544) was developed in a South Brazilian University Hospital. Fifty-one medical students were randomized to ezetimibe 10mg/day or placebo for 5 days. On the fifth and 19th days, blood samples for 25-hydroxycholecalciferol (25OHD), parathyroid hormone (PTH), calcium, and albumin were collected. After the first blood sample collection, all participants received a single oral 50,000 IU cholecalciferol dose during a 15 g-fat meal. Serum 25OHD levels were measured by the immunoassay Diasorin Liaison®. Measurements were compared in a general linear model adjusted for multiple comparisons by the Bonferroni test. Before cholecalciferol administration, 25OHD was <30 ng/mL and <20 ng/mL, respectively, in all and in 82.3% of the participants. Fourteen days after a single 50,000 IU oral dose of cholecalciferol, mean (SD) changes in serum 25OHD were similar in both groups, after adjustment to BMI and 25OHD levels before cholecalciferol administration (p=0.26): 8.7 (3.7) ng/mL in the ezetimibe group, versus 10.0 (3.8) ng/mL in the placebo group. Mean serum 25OHD, PTH, calcium and albumin levels remained similar in both groups. We conclude that ezetimibe had no effect on the mean change in serum 25OHD after a single oral dose of cholecalciferol, in these healthy and young adults.

Asociación de polimorfismos de los genes CRF-BP, SLC6A4 y restricción alimentaria crónica: un estudio preliminar / CRF-BP and SLC6A4 gene polymorphisms among restrained eaters

Background: Restrained eaters (RE) are a group of individuals who constantly restrict their eating. However, they usually alternate restriction with periods of overeating. Aim: To evaluate the possible association of CRF-BP and SLC6A4 gene polymorphisms with chronic alimentary restriction. Material and Methods: The Spanish version of the Revised Restraint Scale was applied to 132 women aged 18 to 25 years. They were divided in a group classified as restrained eaters (RE) and a group of unrestrained eaters. The 5-HTTLPR and CRF-BPs11 polymorphisms of the SLC6A4 and CRF-BP genes were evaluated by polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP), respectively. Results: There was a significant association between the s/s homozygous genotype for the 5-HTTLPR polymorphism of SLC6A4 gene and RE condition (p = 0.033). However, this association was not observed for the CRF-BPs11 polymorphism. Conclusions: The presence of s/s genotype is associated with the RE condition, being the presence of a s allele, a risk factor for this condition.

Multidrug efflux systems in Gram-negative bacteria

Multidrug efflux mechanisms in bacteria contribute significantly to intrinsic and acquired resistance to antimicrobial agents. Genome analysis have confirmed the broad distribution of these systems in Gram-negative as well as in Gram-positive bacteria. Among resistance mechanisms, the multidrug efflux system or pump deserves special attention, since a cell that has acquired it can simultaneously diminish or even suppress the susceptibility to a wide range of antimicrobials. The efflux system is mediated by transport proteins which confer resistance to toxic compounds. In Gram-negative bacteria, a tripartite efflux system is necessary to expel the drug to the outer medium: a protein localized in the cytoplasmic membrane; another in the periplasmatic space (membrane fusion protein - MFP); and a third in the outer membrane (outer membrane factor - OMF). The drug transport is active, and depends either on the energy provided by ATP hydrolysis or is directly driven by the proton motive force. The transport proteins are grouped in families, according to the homology of the amino acid sequences and to similarity of mechanisms. Among Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa have most of the hitherto identified and studied multidrug efflux systems.

Multidrug efflux systems in Gram-negative bacteria

Multidrug efflux mechanisms in bacteria contribute significantly to intrinsic and acquired resistance to antimicrobial agents. Genome analysis have confirmed the broad distribution of these systems in Gram-negative as well as in Gram-positive bacteria. Among resistance mechanisms, the multidrug efflux system or pump deserves special attention, since a cell that has acquired it can simultaneously diminish or even suppress the susceptibility to a wide range of antimicrobials. The efflux system is mediated by transport proteins which confer resistance to toxic compounds. In Gram-negative bacteria, a tripartite efflux system is necessary to expel the drug to the outer medium: a protein localized in the cytoplasmic membrane; another in the periplasmatic space (membrane fusion protein - MFP); and a third in the outer membrane (outer membrane factor - OMF). The drug transport is active, and depends either on the energy provided by ATP hydrolysis or is directly driven by the proton motive force. The transport proteins are grouped in families, according to the homology of the amino acid sequences and to similarity of mechanisms. Among Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa have most of the hitherto identified and studied multidrug efflux systems.