Cytotoxic screening of plants from the Brazilian Atlantic Forest has led to the identification of Casearia arborea and Sorocea hilarii as sources of antitumor compounds.

In the present study, we have evaluated the cytotoxic activity of 282 extracts from 72 native plant species of the Brazilian Atlantic Forest biome. As a result, Casearia arborea and Sorocea hilarii leaves extracts showed cytotoxic activity against three tumour cell lines tested (B16F10, SW480 and Jurkat). After bioassay-guided fractionation, the bioactive fractions were submitted to the dereplication study via High-performance Liquid Chromatography, connected to High-resolution Mass Spectrometry (HPLC-ESI-QTOF/MS) analysis, combined with a Global Natural Products Social Molecular Networking (GNPS) tool. A combination of bioactivity-guided and dereplication approaches resulted in the putative annotation of 27 clerodane diterpenes and 9 flavonoids as main compounds present in the cytotoxic fractions of C. arborea. Regarding the active fraction of S. hilarii, 10 megastigmans, 17 spirostane steroids derivatives and 2 lignans were putatively identified. In conclusion, Casearia arborea and Sorocea hilarii are potential sources of antitumor compounds.

Enteric viruses' dissemination in a private reserve of natural heritage.

This study aimed to assess anthropogenic impact of surrounding population in the Private Reserve of Natural Heritage at Pantanal, the world's largest freshwater wetland ecosystem located in the centre of South America. Viral aetiological agents of acute gastroenteritis as rotavirus A (RVA), noroviruses, human adenoviruses, klassevirus and of hepatitis, as hepatitis A virus, were investigated in different aquatic matrices. Annual collection campaigns were carried out from 2009 to 2012, alternating dry and rainy seasons. Viral particles present in the samples were concentrated by the adsorption-elution method, with negatively charged membranes, and detected by qualitative and quantitative PCR. From a total of 43 samples at least one virus was detected in 65% (28) of them. Viruses were detected in all matrices with concentrations ranging from 2 × 102 to 8·3 × 104 genome copies per litre. A significant higher RVA frequency was observed in the dry season. Our data revealing dissemination of human enteric viruses in water matrices both inside and outside the reserve could be useful to trace faecal contamination in the environment and to minimize the risk of infection by exposure of susceptible individuals. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is part of a collaborative project designed to investigate the environmental and health conditions of the Private Reserve of Natural Heritage at Pantanal, the largest seasonally flooded wetland in the world. The project aimed to promote health and quality of human and wildlife extending technical-scientific knowledge about pathogens present in the region. By assessing the occurrence of human enteric viruses in different water matrices we demonstrated the anthropogenic impact of surrounding population and pointed out the potential risk of infection by exposure of susceptible individuals.

Human enteric viruses in a wastewater treatment plant: evaluation of activated sludge combined with UV disinfection process reveals different removal performances for viruses with different features.

This study assess the quality of wastewater through the detection and quantification of important viruses causing gastroenteritis at different stages of the wastewater treatment process in an activated-sludge wastewater treatment plant with ultraviolet disinfection. Ten sampling events were carried out in a campaign along a period of 18 months collecting wastewater samples from the influent, after the activated-sludge treatment, and after the final disinfection with UV radiation. Samples were concentrated through ultracentrifugation and analysed using retro-transcription, PCR and real time quantitative PCR protocols, for detection and quantification of Group A Rotavirus (RVA), Human Astrovirus (HAstV), Norovirus Genogroup II (NoV GII) and Human Adenovirus (HAdV). HAdV (100%), NoV GII (90%), RVA (70%) and HAstV (60%) were detected in influent samples with concentration from 1·4 (NoV GII) to 8·0 (RVA) log10  gc l-1 . Activated-sludge treatment reached well quality effluents with low organic material concentration, although nonstatistical significant differences were registered among influent and postactivated sludge treatment samples, regarding the presence and concentration for most viruses. All post-UV samples were negative for NoV GII and HAstV, although RVA and HAdV were detected in 38% and 63% of those samples respectively, with concentration ranging from 2·2 to 5·5 and 3·1 to 3·4 log10  gc l-1 . SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates that an activated-sludge wastewater treatment plant with UV disinfection reduces to levels below the detection limit those single-stranded RNA viruses as noroviruses and astroviruses and reach significant lower levels of rotaviruses and adenoviruses after the complete treatment process.

Cerebral blood flow and vasoreactivity in aging: an arterial spin labeling study.

Regional cerebral blood flow (CBF) and cerebrovascular reactivity (CVR) in young and elderly participants were assessed using pulsed arterial spin labeling (ASL) and blood oxygenation level-dependent (BOLD) magnetic resonance imaging (MRI) techniques in combination with inhalation of CO2. Pulsed ASL and BOLD-MRI were acquired in seventeen asymptomatic volunteers (10 young adults, age: 30±7 years; 7 elderly adults, age: 64±8 years) with no history of diabetes, hypertension, and neurological diseases. Data from one elderly participant was excluded due to the incorrigible head motion. Average baseline CBF in gray matter was significantly reduced in elderly (46±9 mL·100 g-1·min-1) compared to young adults (57±8 mL·100 g-1·min-1; P=0.02). Decreased pulsed ASL-CVR and BOLD-CVR in gray matter were also observed in elderly (2.12±1.30 and 0.13±0.06 %/mmHg, respectively) compared to young adults (3.28±1.43 and 0.28±0.11 %/mmHg, respectively; P<0.05), suggesting some degree of vascular impairment with aging. Moreover, age-related decrease in baseline CBF was observed in different brain regions (inferior, middle and superior frontal gyri; precentral and postcentral gyri; superior temporal gyrus; cingulate gyri; insula, putamen, caudate, and supramarginal gyrus). In conclusion, CBF and CVR were successfully investigated using a protocol that causes minimal or no discomfort for the participants. Age-related decreases in baseline CBF and CVR were observed in the cerebral cortex, which may be related to the vulnerability for neurological disorders in aging.

Strychnos pseudoquina A. St. Hil.: a Brazilian medicinal plant with promising in vitro antiherpes activity.

AIMS: To investigate the anti-HSV and anti-inflammatory effects of a standardized ethyl acetate extract (SEAE) prepared with the stem bark of Strychnos pseudoquina, along with two isolated compounds: quercetin 3-O-methyl ether (3MQ) and strychnobiflavone (SBF). METHODS AND RESULTS: The mechanisms of action were evaluated by different methodological strategies. SEAE and SBF affected the early stages of viral infection and reduced HSV-1 protein expression. Both flavonoids elicited a concentration-dependent inhibition of monocyte chemoattractant protein-1 (MCP-1), whereas 3MQ reduced the chemokine release more significantly than SBF. Conversely, both compounds stimulated the production of the cytokines TNF-α and IL-1-ß in LPS-stimulated cells, especially at the intermediate and the highest tested concentrations. CONCLUSIONS: SEAE and SBF interfered with various steps of HSV replication cycle, mainly adsorption, postadsorption and penetration, as well as with ß and γ viral proteins expression; moreover, a direct inactivation of viral particles was observed. Besides, both flavonoids inhibited MCP-1 selectively, a feature that may be beneficial for the development of new anti-HSV agents. SIGNIFICANCE AND IMPACT OF THE STUDY: The results indicated that the samples present anti-HSV and anti-inflammatory activities, at different levels, which is an interesting feature since cold and genital sores are accompanied by an inflammation process.

Norovirus molecular detection in Uruguayan sewage samples reveals a high genetic diversity and GII.4 variant replacement along time.

AIMS: To determine the prevalence and molecular epidemiology of norovirus (NoV) genogroup I (GI) and GII in Uruguay. METHODS AND RESULTS: One hundred and sixteen sewage samples were collected in six cities (Bella Unión, Salto, Paysandú, Fray Bentos, Melo and Treinta y Tres) from March 2011 to April 2013, viruses were concentrated by ultracentrifugation and NoV studies were performed by semi-nested RT-PCR (partial capsid region). NoV were detected in samples from all the cities and detected in 72% (84/116) of the samples with nine of them belonging to GI, 48 to GII and 27 to both genogroups. Remarkably, a high genetic diversity was identified: GII.2 (n = 13), GII.4 (n = 13), GI.1 (n = 5), GI.4 (n = 5), GI.8 (n = 4), GII.13 (n = 4), GII.1 (n = 3), GII.6 (n = 3), GI.3 (n = 1), GI.5 (n = 1), GI.6 (n = 1), GII.3 (n = 1), GII.17 (n = 1). Interestingly, a complete replacement of GII.4 New Orleans 2009 by GII.4 Sydney 2012 variants during 2012 was evidenced. CONCLUSION: This study reveals a high circulation of different NoV GI and GII genotypes in sewage evidencing a replacement of GII.4 variants. SIGNIFICANCE AND IMPACT OF STUDY: This approach can be used as an indicator of the presence of a new GII.4 variant which can originate an increase in acute gastroenteritis outbreaks worldwide.

Sewage surveillance reveals the presence of canine GVII norovirus and canine astrovirus in Uruguay.

Canine norovirus (NoV) and astrovirus (AstV) were studied in 20 domestic sewage samples collected in two cities in Uruguay. Four samples were characterized as canine AstV after phylogenetic analysis clustering with strains detected in Italy and Brazil in 2008 and 2012, respectively. One sample was characterized as canine NoV and clustered with a strain detected in Hong Kong and recently classified as GVII. This study shows the occurrence of a canine NoV GVII strain for the first time in the American continent and also warns about possible zoonotic infection, since canine strains were detected in domestic sewage.

Environmental assessment reveals the presence of MLB-1 human astrovirus in Uruguay.

AIMS: The aim of this study was to investigate the presence of the recently identified human astrovirus (HAstV) and to increase the knowledge of the molecular epidemiology of classical HAstV detected in Uruguay. METHODS AND RESULTS: Recently identified and classical HAstV genotypes were investigated by RT-PCR targeting the ORF1b and ORF2 genome regions in 20 samples obtained between September 2011 and April 2013 in two cities of the eastern region of Uruguay. Four of 20 samples (20%) were identified as MLB-1 genotype and it was found a new MLB-1 classification through the segregation of the worldwide reported MLB-1 strains in two genetic lineages proposed and named: MLB-1a and MLB-1b. Fourteen (70%) samples were positive for classical HAstV and 12 of them were successfully sequenced and genotyped as: HAstV-1 (n = 10), HAstV-2 and HAstV-5 (one sample each). CONCLUSION: These results constitute the first report in the Latin American region concerning the molecular detection and characterization of MLB-1 HAstV strains in environmental samples. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights the benefits of an environmental surveillance to study emerging enteric viruses circulating in human societies.

Environmental Assessment of Classical Human Astrovirus in Uruguay.

The aim of this study was to determine the molecular epidemiology of classical human astrovirus (HAstV) strains in sewage samples from four Uruguayan cities: Bella Unión, Salto, Paysandú, and Fray Bentos, located in the Northwestern region of the country. Overall, 96 sewage samples were collected biweekly between March 2011 and February 2012 and were subject to ultracentrifugation methodology in order to concentrate the viruses. RT-PCR directed to the ORF2 genome region was performed followed by sequencing and phylogenetic analysis. Forty-three (45 %) out of 96 analyzed samples were positive for HAstV (Mamastrovirus 1) and 31 of them were successfully sequenced being 21 (49 %) of them classified as HAstV-1 genotype (1a lineage) and 10 (23 %) as HAstV-2 genotype (eight strains belonging to the 2d lineage and two strains to the 2c lineage). The 1a lineage circulated throughout the year, while the 2d lineage only in the coldest months (June to October). Strikingly, the 2c lineage was detected only in Salto city during March 2011. In this city it was observed the highest frequency of HAstV and the greatest genetic diversity, probably due to its role as high touristic spot with an important influx of visitants from others regions of Uruguay and also from other countries. This study constitutes the first report in Uruguay that describes the phylogenetic diversity and genotype distribution of HAstV strains circulating in the Northwestern region evidencing a high frequency and also the presence of several different lineages.

Phytocannabinoids and epilepsy.

WHAT IS KNOWN AND OBJECTIVE: Antiepileptic drugs often produce serious adverse effects, and many patients do not respond to them properly. Phytocannabinoids produce anticonvulsant effects in preclinical and preliminary human studies, and appear to produce fewer adverse effects than available antiepileptic drugs. The present review summarizes studies on the anticonvulsant properties of phytocannabinoids. METHODS: Literature search using the PubMed database to identify studies on phytocannabinoids and epilepsy. RESULTS AND DISCUSSION: Preclinical studies suggest that phytocannabinoids, especially cannabidiol and cannabidivarin, have potent anticonvulsant effects which are mediated by the endocannabinoid system. Human studies are limited in number and quality, but suggest that cannabidiol has anticonvulsant effects in adult and infantile epilepsy and is well tolerated after prolonged administration. WHAT IS NEW AND CONCLUSION: Phytocannabinoids produce anticonvulsant effects through the endocannabinoid system, with few adverse effects. Cannabidiol and cannabidivarin should be tested in randomized, controlled clinical trials, especially in infantile epileptic syndromes.

Overexpression of the activated form of the AtAREB1 gene (AtAREB1ΔQT) improves soybean responses to water deficit.

Abscisic acid-responsive element binding protein (AREB1) is a basic domain/leucine zipper transcription factor that binds to the abscisic acid (ABA)-responsive element motif in the promoter region of ABA-inducible genes. Because AREB1 is not sufficient to direct the expression of downstream genes under non-stress conditions, an activated form of AREB1 (AREB1ΔQT) was created. Several reports claim that plants overexpressing AREB1 or AREB1ΔQT show improved drought tolerance. In our studies, soybean plants overexpressing AREB1ΔQT were characterized molecularly, and the phenotype and drought response of three lines were accessed under greenhouse conditions. Under conditions of water deficit, the transformed plants presented a higher survival rate (100%) than those of their isoline, cultivar BR 16 (40%). Moreover, the transformed plants displayed better water use efficiency and had a higher number of leaves than their isoline. Because the transgenic plants had higher stomatal conductance than its isoline under well-watered conditions, it was suggested that the enhanced drought response of AREB1ΔQT soybean plants might not be associated with altered transpiration rates mediated by ABA-dependent stomatal closure. However, it is possible that the smaller leaf area of the transgenic plants reduced their transpiration and water use, causing delayed stress onset. The difference in the degree of wilting and percentage of survival between the 35S-AREB1ΔQT and wildtype plants may also be related to the regulation of genes that protect against dehydration because metabolic impairment of photosynthesis, deduced by an increasing internal CO2 concentration, was not observed in the transgenic plants.

Gastroenteric virus dissemination and influence of rainfall events in urban beaches in Brazil.

AIMS: This study was conducted to assess rotavirus A (RV-A), genogroup II (GII) norovirus (NoV), and human adenovirus (HAdV) dissemination in recreational water in an urban beach located in the city of Rio de Janeiro and their persistence during rainfall events. METHODS AND RESULTS: Viruses, including bacteriophage (PP7), used as internal control, were concentrated, reverse transcribed and quantified by a low-cost method based on organic flocculation with skimmed milk coupled with quantitative polymerase chain reaction protocols. The analysis of 74 superficial water samples obtained during 6 months of monitoring detected HAdV (66%), RV-A (37%) and GII NoV (14%), with a mean viral load of 4·1 log10 genome copies l(-1) (g.c. l(-1) ), 4·3 log10 g.c l(-1) and 3·8 log10 g.c. l(-1) , respectively. Investigation of those viruses during two rainfall events showed a longer permanence after rainfall events compared with bacterial indicators. CONCLUSIONS: The results highlight the need for further monitoring using viral parameters to determine the microbiological quality of recreational waters to allow bath in these waters, especially during rainy events. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides data on virus contamination in recreational waters on tourist beaches frequented throughout the year, emphasizing the importance of viral parameters for assessing microbiological quality of water, as well as the potential risk of waterborne infections.

Assessment of gastroenteric viruses from wastewater directly discharged into Uruguay River, Uruguay.

The aim of this study was to assess the viral contamination of group A rotavirus (RVA), norovirus (NoV), and human astrovirus (HAstV) in sewage directly discharged into Uruguay River and to characterize RVA genotypes circulating in Uruguay. For this purpose, sewage samples (n = 96) were collected biweekly from March 2011 to February 2012 in four Uruguayan cities: Bella Unión, Salto, Paysandú, and Fray Bentos. Each sample was concentrated by ultracentrifugation method. Qualitative and quantitative RT-PCR for RVA, NoV, and HAstV were performed. A wide dissemination of gastroenteric viruses was observed in the sewage samples analyzed with 80% of positivity, being NoV (51%) the most frequently detected followed by RVA with a frequency of 49% and HAstV with 45%. Genotypes of RVA were typed using multiplex semi-nested RT-PCR as follows: P[8] (n = 15), P[4] (n = 8), P[10] (n = 1), P[11] (n = 1), G2 (n = 29), and G3 (n = 2). The viral load ranged from 10(3) to 10(7) genomic copies/liter, and they were detected roughly with the same frequency in all participant cities. A peak of RVA and HAstV detection was observed in colder months (June to September), whereas no seasonality was observed for NoV. This study demonstrates for the first time, the high degree of gastroenteric viral contamination in the country; highlighting the importance of developing these analyses as a tool to determine the viral contamination in this hydrographic boundary region used by the local populations for recreation and consumption, establishing an elevated risk of gastroenteric diseases for human health.

Avaliação fitoquímica e atividade antioxidante, antimutagênica e toxicológica do extrato aquoso das folhas de Ocimum gratissimum L / Phytochemical Assessment And Antioxidant, Antimutagenic And Toxicological Activity Of Ocimum gratissimum L. Leaf aqueous extract

A espécie Ocimum gratissimum L., popularmente conhecida como alfavaca, é uma planta muito usada na medicina tradicional brasileira, à qual são atribuídas diversas atividades terapêuticas quando usada na forma de infuso de suas folhas. Neste estudo foi realizada a caracterização fitoquímica, a avaliação da ação antioxidante e a investigação dos efeitos antimutagênico e antigenotóxico, além do efeito mutagênico e genotóxico potencial do extrato aquoso liofilizado a parir das folhas de O. gratissimum (EAOG). O conteúdo de polifenóis totais no extrato foi determinado pelo método Folin-Ciocalteu, sendo encontrado 11,3 µg EAG/mg de EAOG. A atividade antioxidante foi avaliada pelo teste do 1,1-difenil-2-picril hidrazil (DPPH•), apresentando IC50 de 83,0 µg/mL. A antimutagenicidade e mutagenicidade foram avaliadas em cepas de Salmonella typhimurium (TA98 e TA100) utilizando o teste Salmonella/microssoma (Salmonella typhimurium/microssomas) em diferentes concentrações. EAOG induziu a atividade antimutagênica para a cepa TA98. A mutagenicidade não foi observada para o extrato em ambas as linhagens. Adicionalmente, a ação antigenotoxica avaliada pelo teste de clivagem do DNA-plasmidial também foi observada para EAOG. Os resultados também demonstraram que o extrato não foi capaz de induzir a genotoxicidade pelo teste empregado. Este estudo relata, pela primeira vez, as propriedades antimutagênica e antigenotóxica do extrato aquoso de O. gratissimum.

The species Ocimum gratissimum L., popularly known as Clove Basil, is a plant widely used in traditional Brazilian medicine, and several therapeutic activities are attributed to it when used as infusion of its leaves. In this study, we carried out a phytochemical characterization and the assessment evaluation and investigation of the antioxidant action of the antimutagenic and antigenotoxic effects and the potential mutagenic and genotoxic effects of the freeze-dried aqueous extract of the O. gratissimum (EAOG) leaves. The total polyphenol content in the extract was determined by the Folin-Ciocalteu method, and we found 11.3 µg EAG/mg of EAOG. The antioxidant activity was assessed by the 1,1-diphenyl-2-picryl hidrazil (DPPH·), with IC50 of 83.0 µg/mL. Antimutagenicity and mutagenicity were assessed in Salmonella typhimurium (TA98 and TA100) strains using the Salmonella/microsome (Salmonella typhimurium/microsome) test in different concentrations. EAOG induced antimutagenic activity for strain TA98. Mutagenicity was not observed for the extract in both strains. Additionally, antigenotoxic action, assessed by cleavage of the DNA-damage, was also observed for EAOG. The results also show that the extract was not able to induce genotoxicity by the test used. This study reports for the first time the antimutagenic and antigenotoxic properties of the O. gratissimum aqueous extract.

Fitodefensivos em plantas medicinais: macromoléculas hidrofílicas de folhas de mil folhas (Achillea millefolium L. ) inibem o crescimento in vitro de bactérias fitopatogênicas / Agrochemicals in medicinal plants: hydrophilic macromolecules from leaves of "mil folhas" (Achillea millefolium L. ) inhibit in vitro growth of phytopathogenic bacteria

Extratos aquosos da planta medicinal Achillea millefolium contêm macromoléculas de interesse para desenvolver fitodefensivos para a agricultura. Duas frações de mil folhas foram obtidas por ultrafiltração, E1 (contendo moléculas maiores que 30 kDa), e E3 (peptídeos entre 1 e 10 kDa) que inibiram o crescimento das bactérias fitopatogênicas Ralstonia solanacearum, gram-negativa, e Clavibacter michiganensis subsp. michiganensis, gram-positiva, com dependência de concentração. Os valores de concentração inibitória mínima (CIM) para ambos os extratos e bactérias foram baixos, entre 20 e 80µM. A CIM relativa à proteína total evidenciou a presença de macromoléculas muito ativas em E3, embora com baixa concentração proteica. E3 se aplica à prospecção de peptídeos antimicrobianos. Estimar a CIM relativa à quantidade de amostra vegetal valorizou o potencial antimicrobiano natural de E1, que contém alta concentração proteica. E1e E3 se aplicam ao desenvolvimento de fitodefensivos para uso biotecnológico. A ultrafiltração fracionou as amostras de forma nativa, rápida, e com baixo custo; além de dessalinizar, clarificar, purificar, e concentrar E1 e E3. Esse estudo inédito sobre a separômica e a ação antimicrobiana de extratos macromoleculares aquosos de mil folhas sugere que plantas cicatrizantes podem apresentar grande potencial para desenvolver fitodefensivos agrícolas naturais não danosos, à semelhança de medicamentos fitoterápicos.

Aqueous extracts from the medicinal plant Achillea millefolium contain macromolecules of interest to develop agrochemicals for agriculture. Two fractions of "mil folhas" were obtained by ultrafiltration, E1 (containing molecules larger than 30 kDa) and E3 (peptides between 1 and 10 kDa), which inhibited the growth of phytopathogenic bacteria Ralstonia solanacearum, gram-negative, and Clavibacter michiganensis subsp. michiganensis, gram-positive, concentration-dependent. The values of minimum inhibitory concentration (MIC) for both extracts and both bacteria were low, ranging from 20 to 80µM. The MIC relative to total protein evidenced the presence of very active macromolecules in E3, although showing low protein concentration. E3 applies to the prospection of antimicrobial peptides. The estimated MIC relative to the amount of plant sample valued the natural antimicrobial potential of E1, which contains high protein concentration. E1 and E3 can be used in the development of agrochemicals for biotechnological purposes. The ultrafiltration procedure fractionated the samples in a rapid and native way and at a low cost; it also desalted, clarified, concentrated and purified both E1 and E3. This pioneering study on the separomics and on the antimicrobial activity of macromolecular aqueous extracts from "mil folhas" suggests that healing plants have great potential to develop non-harmful agricultural natural agrochemicals, similarly to the available phytotherapic drugs.

Produtos naturais ativadores de PPAR e marcadores associados ao processo inflamatório na Síndrome Metabólica / The inflammatory process in the metabolic syndrome: PPAR activators of natural products and markers associated with metabolic disorders

O processo inflamatório é o elo entre a síndrome metabólica e as doenças cardiovasculares. Para verificar a presença e o grau da inflamação, vários biomarcadores têm sido propostos e investigados. Este trabalho tem como objetivo revisar as recentes pesquisas que associam alguns marcadores expressos no tecido adiposo, enfatizando, dentre eles, a adiponectina, a resistina, a leptina e o transportador de glicose GLUT-4 na síndrome metabólica, a relação da inflamação decorrente desse conjunto de desordens metabólicas sob os receptores proliferadores peroxissomais (PPARs), bem como o efeito de diferentes extratos vegetais e produtos naturais bioativos na ativação desses receptores.

The inflammatory process is the link between metabolic syndrome and cardiovascular diseases. To verify the presence and degree of inflammation, several biomarkers have been proposed and different receptors have been investigated. This study aims to review recent researches involving some markers expressed in the adipose tissue, emphasizing, among them, adiponectin, resistin, leptin and glucose transporter GLUT-4 in the metabolic syndrome, the relationship of inflammation arising from this set of metabolic disorders on the peroxisome proliferator receptors (PPARs) and the effect of different bioactive compounds in the activation of these receptors.

Analysis of the NMDA in focal cerebral ischemia in rats / Análisis del NMDA en isquemia cerebral focal en ratas

NMDAR (N-methyl-D-aspartate receptor) is one subtype of ionotrophic glutamate receptor which is extensively distributed in the central nervous system (CNS). In the mammalian CNS, NMDAR serves prominent roles in the pathophysiologic process of cerebral ischemia. This study aimed to investigate the pattern of expression of protein and gene of the excitatory neurotransmitter NMDAR in experimental focal cerebral ischemia and the hole of neuroprotection with hypothermia and ketoprofen. 120 rats were randomly divided into 6 groups (20 animals each): control - no surgery; sham - simulation of surgery; ischemic - focal ischemia for 1 hour, without reperfusion; ischemic + intraischemic hypothermia; ischemic + previous intravenous ketoprofen, and ischemic + hypothermia and ketoprofen. Ten animals from each experimental group were used to establish the volume of infarct. Transient focal cerebral ischemia was obtained in rats by occlusion of the middle cerebral artery with an intraluminal suture. The infarct volume was measured using morphometric analysis of infarct areas defined by triphenyl tetrazolium chloride and the patterns of expression of the protein and gene NMDA were evaluated by immunohistochemistry and quantitative real-time PCR, respectively. Increases in the protein and gene NMDA receptor in the ischemics areas were observed and these increases were reduced by hypothermia and ketoprofen. The increase in the NMDA receptor protein and gene expression observed in the ischemic animals was reduced by neuroprotection (hypothermia and ketoprofen). The NMDA receptor increases in the ischemic area suggests that the NMDA mediated neuroexcitotoxicity plays an important role in cell death and that the neuroprotective effect of both, hypothermia and ketoprofen is directly involved with the NMDA...

NMDAR (N-metil-D-aspartato) es un tipo de receptor de glutamato ionotrópico y está ampliamente distribuido en el sistema nervioso central (SNC). En el SNC de mamíferos, NMDAR se destaca de manera importante en el proceso fisiopatológico de la isquemia cerebral. Este estudio tuvo como objetivo investigar el patrón de expresión de proteínas y genes para el NMDA neurotransmisor excitatorio experimental de la isquemia cerebral focal y el vacío en la neuroprotección con hipotermia y ketoprofeno. Se dividieron 120 ratas aleatoriamente en grupos de 6 animales cada uno (20): Control - sin cirugía; Sham - simulación de cirugía; isquémicas - isquemia focal durante 1 hora, sin reperfusión isquémica; hipotermia intra-isquémica; isquemia; previa aplicación de ketoprofeno intravenoso, e hipotermia isquémica y ketoprofeno. Diez animales de cada grupo experimental fueron utilizados para establecer el volumen de infarto.La isquemia cerebral focal transitoria fue obtenida en ratas mediante oclusión de la arteria cerebral media con una sutura intraluminal. El volumen de infarto fue medido mediante análisis morfométrico de las áreas de infarto definidas por cloruro de trifenil tetrazolio y patrones de expresión de la proteína y el gen de NMDA, fueron evaluados por inmunohistoquímica y PCR cuantitativa en tiempo real, respectivamente. Se observaron aumentos en la proteína y en el gen del receptor de NMDA en las áreas isquémicas y estos aumentos fueron reducidos por la hipotermia y ketoprofeno. El aumento de la proteína del receptor de NMDA y la expresión génica observada en los animales isquémicos fue reducido mediante hipotermia y ketoprofeno. Los aumentos del receptor de NMDA en el área isquémica sugiere que la neuro excitotoxicidad mediada por NMDA desempeña un papel importante en la muerte celular y que el efecto neuroprotector de ambos, hipotermia y ketoprofeno está directamente relacionado al NMDA...

Extratemporal damage in temporal lobe epilepsy: magnetization transfer adds information to volumetric MR imaging.

BACKGROUND AND PURPOSE: MTS is characterized by gliosis and atrophy of the hippocampus and related limbic structures. However, the damage is not limited to those structures with atrophy and has been reported in extratemporal regions. Because volumetric studies are nonspecific, the pathophysiology of the brain damage remains to be solved. MTI is an MR imaging technique more sensitive to subtle neuropathologic changes than conventional MR imaging. Here we combined MTI with VBM analysis to evaluate extratemporal damage in patients with TLE. MATERIALS AND METHODS: We studied 23 healthy controls and 21 patients with TLE with mean ages, respectively, of 37.6 ± 10.9 and 38.6 ± 9.02 years. All subjects had a full clinical follow-up and MR imaging. We processed the images with VBM for volumetric analysis of WM and GM, as well as with voxel-based analysis of MTR for macromolecular integrity analysis. RESULTS: In addition to MTR decrease in the temporal lobes, we found a significant decrease in GM and WM volumes. In the WM, the MTR decrease was correlated to volume loss detected by VBM, indicating that brain atrophy may explain part of the MTR decrease. We also found areas in which the MTR decrease was not associated with volume loss, suggesting an additional pathophysiologic process other than neuronal loss and atrophy underlying the MTR changes. CONCLUSIONS: Our results support the hypothesis that there are widespread lesions in the brain, including the corpus callosum and the frontal lobe, affecting both GM and WM.

Monitoring of canine parvovirus (CPV) strains detected in vaccinated puppies in Brazil.

The objective of this study was to investigate, by partial sequencing of VP2 protein, the variability of CPV detected in 37 fecal samples collected from vaccinated puppies with enteritis. Laboratorial diagnosis of CPV was confirmed by HA/HI and PCR and, for sequencing analyses, two different regions of the VP2 gene were amplified by PCR. From 1995 to 2004, all strains were characterized as CPV-2a. After that, both CPV-2a and CPV-2b were detected. All CPV-2a showed a non-synonymous mutation in the residue 297 (Ser→Ala). A synonymous substitution at the AA 574 was also observed in 15/37 samples. Our findings indicate that the cases of vaccine failure are most likely not associated to the mutations detected in the sequenced regions. However, the monitoring of genotyping mutations that led to new CPV strains is essential to determinate if current vaccines will keep providing protection against all new future variants.