Evaluation of enterotoxins and antimicrobial resistance in microorganisms isolated from raw sheep milk and cheese : ensuring the microbiological safety of these products in southern Brazil

This study emphasizes the importance of monitoring the microbiological quality of animal products, such as raw sheep’s milk and cheese, to ensure food safety. In Brazil, there is currently no legislation governing the quality of sheep’s milk and its derivatives. Therefore, this study aimed to evaluate: (i) the hygienic-sanitary quality of raw sheep’s milk and cheese produced in southern Brazil; (ii) the presence of enterotoxins and Staphylococcus spp. in these products; and (iii) the susceptibility of the isolated Staphylococcus spp. to antimicrobial drugs and the presence of resistance genes. A total of 35 samples of sheep’s milk and cheese were examined. The microbiological quality and presence of enterotoxins were accessed using Petrifilm and VIDAS SET2 methods, respectively. Antimicrobial susceptibility tests were conducted using VITEK 2 equipment and the disc diffusion method. The presence of resistance genes tet(L), sul1, sul2, ermB, tetM, AAC(6)’, tetW, and strA were evaluated through PCR. In total, 39 Staphylococcus spp. were obtained. The resistance genes tetM, ermB, strA, tetL, sul1, AAC(6)’, and sul2 were detected in 82%, 59%, 36%, 28%, 23%, 3%, and 3% of isolates, respectively. The findings revealed that both raw sheep’s milk and cheese contained Staphylococcus spp. that exhibited resistance to antimicrobial drugs and harbored resistance genes. These results underscore the immediate need for specific legislation in Brazil to regulate the production and sale of these product

Epidemiologic and clinical characteristics of pregnant women living with HIV/AIDS in a region of Southern Brazil where the subtype C of HIV-1 infection predominates

AbstractSouthern Brazil has the highest prevalence rate of AIDS in the country and is the only region in the Americas where HIV-1 subtype C prevails.ObjectiveWe evaluated the epidemiologic and clinical characteristics of pregnant women living with HIV/AIDS in the South region of Santa Catarina, Brazil.MethodsAll pregnant women with HIV infection attending the obstetric outpatient clinic of Criciúma, State of Santa Catarina, in 2007 (n=46) were invited to participate. Data of 36 eligible participants were obtained through a standardized questionnaire.ResultsThe great majority were young, with a steady partner, low family income, low education level and referring early first sexual intercourse. Many reported use of illicit non-injecting drugs (55.5%) and unprotected sex with partners that were HIV-positive (57.7%), injecting drug user (22.2%), male inmate (19.4%), truck driver (13.8%), with history of sexually transmitted disease (11.1%) or men who have sex with men (MSM) (2.8%). Most (66.7%) of the participants had their HIV diagnosis done during the pregnancy, 7 (19.4%) had a previous history of HIV mother-to-child transmission. Therapy based on highly active antiretroviral therapy (94%) was initiated at 19.3 weeks on average and 33% showed irregular antiretroviral adherence.ConclusionThese results confirm previous data on HIV epidemiology in Brazil and suggest that the women partners’ sexual behavior and unprotected sexual intercourse are important aspects of HIV epidemic. Additional efforts in education, prophylaxis and medication adherence are needed

Evaluation of the genotoxic and antigenotoxic potential of Melissa officinalis in mice

Melissa officinalis (L.) (Lamiaceae), a plant known as the lemon balm, is native to the east Mediterranean region and west Asia. Also found in tropical countries, such as Brazil, where it is popularly known as “erva-cidreira” or “melissa”, it is widely used in aqueous- or alcoholic-extract form in the treatment of various disorders. The aim was to investigate in vivo its antigenotoxicity and antimutagenicity, as well as its genotoxic/mutagenic potential through comet and micronucleus assaying. CF-1 male mice were treated with ethanolic (Mo-EE) (250 or 500 mg/kg) or aqueous (Mo-AE) (100 mg/kg) solutions of an M. officinalis extract for 2 weeks, prior to treatment with saline or Methyl methanesulfonate (MMS) doses by intraperitoneal injection. Irrespective of the doses, no genotoxic or mutagenic effects were observed in blood and bone-marrow samples. Although Mo-EE exerted an antigenotoxic effect on the blood cells of mice treated with the alkylating agent (MMS) in all the doses, this was not so with Mo-AE. Micronucleus testing revealed the protector effect of Mo-EE, but only when administered at the highest dose. The implication that an ethanolic extract of M. officinalis has antigenotoxic/antimutagenic properties is an indication of its medicinal relevance

Lithium response in bipolar disorder - study of the molecular response variability in lymphoblastoid cell-lines from excellent-responders and non-responders

Les troubles bipolaires (TBs) (2% de la population générale) sont une maladie psychiatrique chronique caractérisée par des fluctuations répétées entre des épisodes d'humeur maniaque/hypomaniaque ou mixte et des épisodes dépressifs et ayant un impact sociétal considérable. Les sels de lithium (Li) sont le choix thérapeutique de première intention dans la prévention des rechutes lors du traitement des TBs. Néanmoins les taux de réponse restent très variables et actuellement aucun biomarqueur prédictif de la réponse n'est disponible en pratique clinique. Dans cette étude à l'aide d'un modèle de lignées lymphoblastoïdes (LLB) ; provenant de patients Caucasiens atteints par un TB et caractérisés pour leur réponse prophylactique au Li (échelle Alda) en tant que excellents-répondeurs (ER) et non-répondeurs (NR) au traitement ; nous avons analysé sur des puces pangénomiques et par RT-qPCR les effets du LiCl (1 mM) sur le transcriptome (gènes et microARNs) et sur le profil d'activité fonctionnelle de serine-theronine kinases. Sur une approche in silico, des réseaux d'interaction (gènes/microARNs) basés sur les résultats des analyses transcriptomiques ont été construits afin de mieux appréhender la réponse moléculaire au Li entre ERs et NRs. Des approches gène-candidat ont aussi été conduites au sein de cette étude. Dans l'approche ciblée, nous avons étudié les niveaux d'expression du gène GADL-1, un gène-candidat présent dans la littérature et pour lequel une forte association entre ses polymorphismes et la réponse au Li avait été démontrée. Au sein de nos échantillons aucune différence significative n'a été trouvée au niveau transcriptomique entre ER, NR et sujets contrôles. L'approche pangénomique indique que le Li possède des effets distincts sur les profils d'expression des gènes et microARNs dans les lignées d'ERs et NRs, notamment avec très peu de transcrits candidats en commun. L'analyse in silico a démontré que les réseaux associés au groupe ER possèdent moins des connexions, ce qui suggère que le phénotype de réponse dans le groupe ER repose sur un nombre réduit de gènes ou un moindre degré d'hétérogénéité. Par ailleurs, les réseaux obtenus dans le groupe NRs présentent un plus grand nombre de noeuds et des gènes, ce que suggère un effet plus diffus du Li chez les NRs comparé aux ERs. Nous avons ainsi identifié des gènes centraux dans les réseaux d'interaction au sein du groupe ER (NTRK1, NEDD4, ERBB2) et NR (UBC, MAPK8, INS). Enfin, les résultats sur les profils kinomiques semblent indiquer que le LiCl (1 mM) ne modulerait pas de façon différentielle l'activité des kinases dans les groupes ER et NR. Néanmoins, lors de l'analyse des profils kinomiques en absence de Li, une tendance a été trouvée, ce qui indiquerait que l'activité basale des kinases pourrait être distincte entre les groupes ER et NR. Ces résultats permettent d'élaborer de nouvelles hypothèses sur les voies de signalisations et réseaux d'interaction de protéines associés à la réponse thérapeutique au Li. Cependant, la validation et la réplication de ces résultats dans d'autres conditions (e.g.augmentation des effectifs, cellules mononuclées du sang..) sont nécessaires. Ces travaux permettent de souligner l'intérêt des approches multidimensionnelles (génétiques, épigénétiques, transcriptionnelles et fonctionnelles) dans la recherche de biomarqueurs de la réponse au Li dans le TBBipolar disorder (BD) (2% of the general population) is a major affective disorder characterized by recurrent manic and depressive episodes and associated with considerable burden and costs. Lithium (Li) is the leading treatment for relapse prevention in BD, although the level of response is highly variable and no biomarker to indicate clinical utility is currently available. Using bipolar lymphoblastoid cell-lines (LCL) from Caucasian BD patients, selected for their excellent (ER) or poor (NR) prophylactic Li response (Alda scale) to lithium; we assessed by microarrays and RT-qPCR the effects of LiCl (at 1 mM) on the transcriptome (genes and microRNAs) and at the functional activity profile of serine-threonine kinases. In silico network approaches based on the transcriptomic results (gene/microRNAs interactions) were conducted to better characterize the molecular response to Li between ER and NR. Individual gene-candidate approaches were also explored. In the targeted approach, we compared the expression levels of GADL-1 gene, a candidate-gene presenting polymorphisms highly associated with response to lithium. In our sample we did not find any significant differences between ER, NR and healthy volunteers. In the whole genome approach, we found that lithium induces distinct effects of on the gene and miRNA expression profiles in ER and NR cell-lines, with few candidates shared by both groups. Our network analysis reveals that ER networks exhibits fewer connections, which may indicate that the Li-response phenotype relies on fewer genes or a lower heterogeneity in ER samples. On the other hand, NR networks show a higher number of nodes and genes, suggesting a more diffuse Li-effect in NR than in ER. With this approach we were able to identify hub genes candidates in the ER (NTRK1, NEDD4, ERBB2) and NR groups. Finally, the results from the kinome profiling, seem to indicate that lithium at 1mM does not modulate differentially kinase activity in ER and NR groups. However, a trend was found when comparing baseline profile (i.e. in the absence of lithium). These data suggest that basal kinases activities are different between ER and NR phenotypes. These results allow to develop new hypotheses on the signaling pathways and interaction networks of proteins associated with the therapeutic response to Li. However, validation and replication of these results under other conditions (number increase, peripheral blood mononuclear cells...) are necessary. This work highlights the value of multidimensional approaches (genetic, epigenetic, transcriptional and functional) in the search for biomarkers of the response to Li in BD

Lithium response in bipolar disorder: No difference in GADL1 gene expression between cell lines from excellent-responders and non-responders

International audienc

Lithium effects on serine-threonine kinases activity: High throughput kinomic profiling of lymphoblastoid cell lines from excellent-responders and non-responders bipolar patients

International audienc

Lithium response in bipolar disorders and core clock genes expression

<p><b>Objectives:</b> We examine whether the lithium response is associated with changes in the expression of core clock genes.</p> <p><b>Methods:</b> The effect of a therapeutic concentration of lithium (1 mM) on the expression levels of 17 circadian genes was examined in lymphoblastoid cell lines (LCLs) derived from two well-characterized groups of bipolar disorder patients, defined as lithium non-responders (NR, <i>n</i> = 20) or excellent responders (ER, <i>n</i> = 16). Quantitative real-time PCR (qRT-PCR) was conducted at 2, 4 and 8 days (d2, d4 and d8) with and without lithium exposure.</p> <p><b>Results:</b> At d2, in ER only, <i>BHLHE41</i>, <i>RORA</i>, <i>PER1</i>, <i>ARNTL</i>, <i>CRY2</i>, <i>BHLHE40</i> and <i>CSNK1D</i> were upregulated, whereas <i>NR1D1</i> was downregulated. At d4, in ER only, <i>CRY1</i> was downregulated. At d8, in NR only, <i>GSK3β</i> was upregulated and <i>DBP</i>, <i>TIMELESS</i> and <i>CRY1</i> were downregulated. Significant Group × Lithium interactions existed for <i>NR1D1</i> at d2 (<i>P</i> = 0.02), and <i>CRY1</i> at d4 (<i>P</i> = 0.02). Longitudinal analyses showed differential temporal evolutions between NR and ER (significant Time × Group interaction) for <i>PER3</i>, <i>NR1D1</i>, <i>DBP</i>, <i>RORA</i>, <i>CSNK1D and TIMELESS</i>; and a significant Time × Lithium interaction for <i>NR1D1</i>. Coexpression data analyses suggested distinct groups of circadian genes concurrently modulated by lithium.</p> <p><b>Conclusions:</b> In LCLs, lithium influences expression of circadian genes with differences in amplitude and kinetics according to the patient’s lithium response status.</p