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Leishmaniasis are endemic diseases caused by different species of intracellular parasites of the genus

Introduction: Carriers of oncohematological diseases are at high risk for hepatitis B virus (HBV) infection. Objective and method: To investigate the epidemiology of HBV infection in Goiânia, Central Brazil, 322 individuals with oncohematological diseases (leukemias, Hodgkin lymphoma and non-Hodgkin lymphoma) were interviewed and blood samples were collected for the detection of serological markers of HBV-DNA by polymerase chain reaction (PCR). Medical records of participants were also reviewed. Results: Non-Hodgkin’s lymphomas (n = 99) and chronic myeloid leukemia (n = 108) were the most frequent oncohematological diseases. The overall prevalence of HBV was 13.97% (45/322). Of the total participants, 8.69% (28/322) presented isolated positivity for anti-HBs, suggesting low vaccine coverage. HBV-DNA was detected in 25% (1/4) of HBsAg positive samples and in 25% (3/12) of anti-HBc isolated, suggesting HBV occult infection. All samples were identified as subgenotype A1. Entries in patient records and the findings of this investigation suggest anti-HBc seroconversion during oncologic treatment. Age 50 years or over and use of a central catheter during therapy were associated with HBV exposure. Conclusion: The low frequency of hepatitis B immunized individuals, detection of HBV DNA in HBsAg negative samples, and the suggestion of HBV exposure during treatment evidenced the potential for health-related viral dissemination in people with oncohematological diseases in our region, reinforcing the importance of serological monitoring, vaccination against hepatitis B, and adoption of strict infection control measures in these individuals. Keywords: Epidemiology, Hepatitis B, Leukemia, Lymphoma, Oncology

Anthropogenic actions, including deforestation, disorganized urbanization, and globalization, contribute to emergence and reemergence of arboviruses worldwide, where Flavivirus is the most prevalent, and its continuous monitoring can help in preventive control strategies. Thus, the aim of this study was to detect flavivirus RNA in single hematophagous insects, which are used as sentinels. Total RNA was extracted from six Aedes aegypti stored since 2003 and from 100 Culicidae and collected through CDC trap in a public park of a Brazilian Northwest city of São Paulo State. Flavivirus was detected through RT/PCR targeting 230–250 bp of the RNA polymerase coding sequence (NS5). PCR amplicons were sequenced by Sanger method, used in comparative analysis over Basic Local Alignment Search Tool (BLAST) in GenBank, and subjected to Neighbor-Joining phylogenetic analyses. Efficiency of Flavivirus diagnosis was confirmed by detection of Dengue virus serotype 2 in Ae. aegypti. From the 100 collected insects, 19 were positive for Culex flavivirus (CxFV). NS5 partial sequence phylogenetic analysis clustered all CxFV in one branch separated from vertebrate flaviviruses, being applicable to the identification of Flavivirus species. The dipteran RNA extraction methodology described in this work supports detection of flaviviruses in single insects maintained in 80% ethanol, which can be used to constant arbovirus surveillance.

Orientadora: Profa. Dra. Márcia Aparecida Sperança Dissertação (mestrado) - Universidade Federal do ABC, Programa de Pós-Graduação em Evolução e Diversidade, Santo André, 2019. As arboviroses estão distribuídas mundialmente, com mais de 390 milhões de casos humanos por ano. Especialmente em regiões tropicais e subtropicais, onde a presença de seus vetores está consolidada devido ao clima favorável, os arbovírus são responsáveis por grandes desafios para a saúde pública dos países afetados, como o Brasil. O gênero Flavivirus compreende os arbovírus mais prevalentes, dentre eles os vírus da dengue, da Zika e da febre amarela. De acordo com o Ministério da Saúde, entre 2016 e 2017, ocorreram 1.732.679 casos de dengue, com 864 óbitos; 233.800 casos de febre Zika, com 08 mortes; e 1.908 casos de febre amarela, com 548 mortes. Além da perda de vidas humanas, há grandes impactos econômicos e sociais envolvidos. O monitoramento das ocorrências de flavivírus durante períodos interepidêmicos pode ajudar as instituições de saúde a desenvolver estratégias de controle preventivo. Assim, o objetivo deste estudo foi detectar a ocorrência de flavivírus diretamente em insetos hematófagos coletados em ambientes naturais, de forma a preservar o espécime, permitindo sua identificação para estudos filogenéticos e ecológicos. Nesta pesquisa, foram processados 102 insetos hematófagos coletados na cidade de Marília, estado de São Paulo. Os insetos foram incubados em um tampão de lise com proteinase K, para liberação de ácidos nucleicos e em seguida o RNA foi purificado. A detecção dos flavivírus foi realizada por transcrição reversa e reação em cadeia da polimerase, utilizando os oligonucleotídeos cFD2 e MAMD, complementares à proteína não-estrutural 5 (NS5) do gênero Flavivirus. O fragmento de 220-250 pb foi obtido em 19 amostras de mosquitos, e a análise por "Basic Local Alignment Search Tool - BLAST" dos dados de sequenciamento dessas dezenove amostras apresentou similaridade de 94-96% com Culex Flavivirus. Sabendo que os mosquitos vetores podem se alimentar de sangue infectado e que podem transmitir vírus durante todo o seu ciclo de vida, esses mosquitos podem ser usados como indicadores epidemiológicos (sentinelas) da circulação de arbovírus em uma determinada área. Arboviruses are distributed worldwide, with more than 390 million human cases per year. Especially in tropical and subtropical regions, where the presence of their vectors is consolidated due to the favorable climate, arboviruses are responsible for major public health challenges in affected countries, such as Brazil. The genus Flavivirus comprises the most prevalent arboviruses, among them the dengue virus, Zika virus and yellow fever virus. According to the Ministry of Health, between 2016 and 2017, there were 1,732,679 cases of dengue, with 864 deaths; 233,800 cases of Zika fever, with 8 deaths; and 1,908 cases of yellow fever, with 548 deaths. In addition to the loss of human lives, there are major economic and social impacts involved. Monitoring of occurrences of flaviviruses during interepidemic periods may help health institutions develop preventive control strategies. Thus, the objective of this study was to detect the occurrence of flavivirus directly in hematophagous insects collected in natural environments, in order to preserve the specimen, allowing its identification for phylogenetic and ecological studies. In this research, 102 hematophagous insects were collected in the city of Marília, state of São Paulo. The insects were incubated in a lysate buffer with proteinase K, to release nucleic acids and then the obtained RNA was purified. The detection of flavivirus was performed by reverse transcription and polymerase chain reaction, using the cFD2 and MAMD oligonucleotides, complementary to the nonstructural protein 5 (NS5) of the genus Flavivirus. The 220-250 bp fragment was obtained from 19 mosquito samples, and the Basic Local Alignment Search Tool - BLAST analysis of the sequencing data from these nineteen samples showed 94-96% similarity with Culex Flavivirus. Knowing that vector mosquitoes can feed on infected blood and can transmit viruses throughout their life cycle, these mosquitoes can be used as epidemiological indicators (sentinels) of the arbovirus circulation in a given area.

Infection control measures have been responsible for a decline in the prevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections in hemodialysis patients. In Brazil, these measures have been in place since 1996. The aim of this study was to evaluate the current HBV and HCV epidemiology among hemodialysis patients in the State of Tocantins comparing them with those found 14 years ago. There was a significant decline in hepatitis B surface antigen (HBsAg) and anti-HCV prevalence from 4% and 13% in 2001 to 0.8% and 2.8% in 2014-2015, respectively (p < 0.05). Variables related to hemodialysis environment such as working shift and length of time on hemodialysis treatment were no longer associated to HCV and HBV exposure in 2014-2015. A high prevalence of self-reported hepatitis B vaccination was observed in both periods, but only 30% of the individuals showed serological profile of effective previous immunization, suggesting a low compliance with surveillance of hepatitis B immunization in hemodialysis centers. The significant decline in viral hepatitis B and C prevalence in hemodialysis patients in Tocantins underscores the importance of infection control measures, but the low frequency of protective serological profile after immunization against hepatitis B points to the need for greater vigilance of the patients’ vaccination.

To date, a tissue depletion study of moxidectin (MOX) in lambs is not available. Thus, considering that lamb meat is of great commercial interest in the world, the aim of the present study was to determine the residue levels of MOX in lamb target-tissues (muscle, liver, kidney and fat) and subsequently calculate the MOX withdrawal period. For this purpose, the target-tissues were analysed by ultra-high-performance liquid chromatography-tandem mass spectrometry. Method validation was performed based on Commission Decision 2002/657/EC and VICH GL49. To quantify the analyte, matrix-matched analytical curves were constructed with spiked blank tissues. The limits of detection and quantitation were 1.5 and 5 ng g−1, respectively, for all matrices. The linearity, decision limit, detection capability accuracy and inter- and intra-day precision of the method are reported. The lambs were treated with a single subcutaneous dose of 0.2 mg MOX kg−1 body weight and were slaughtered in accordance with accepted animal care protocols. Samples of target-tissues were collected on 2, 4, 7, 14, 28 and 42 days after MOX administration. During the whole study, the highest drug residue level occurred in the fat. For the other target-tissues (muscle, liver and kidney), MOX concentrations were below the maximum residue limit (MRL). Considering the MRL value of 500 µg kg−1 for MOX residues in sheep fat, our results in lambs allowed the estimation of a MOX withdrawal period of 31 days. This indicates that the withdrawal period established for MOX in adult sheep (28 days) does not apply for lambs.

Oral anticoagulants are among the drugs with the greatest number of drug interactions. The concomitant use of several medications is a common practice in patients with cardiovascular problems, who often also present with depression; therefore, the probability of an interaction occurring between warfarin and the antidepressants is high, and may result in increased or decreased anticoagulant activity. Since the possible interactions between these two classes of drugs have been poorly explored in literature, with a risk to the patients who use them, we reviewed the pharmacology of warfarin and its possible interactions with antidepressants. Of the antidepressants analyzed, those that showed relevant effects on the interaction with warfarin were, in decreasing order: paroxetine, venlafaxine, fluoxetine, and duloxetine. Os anticoagulantes orais estão entre as drogas com maior número de interações medicamentosas. O uso concomitante de vários medicamentos é uma prática comum em pacientes com problemas cardiovasculares, os quais frequentemente também apresentam depressão; assim, a probabilidade de ocorrer alguma interação entre a varfarina e os antidepressivos é bem expressiva, podendo resultar em um aumento ou uma diminuição da atividade anticoagulante. Como as possíveis interações entre essas duas classes de medicamentos se mostraram pouco exploradas na literatura, com risco aos pacientes que fazem uso delas, revisamos a farmacologia da varfarina e suas possíveis interações com antidepressivos. Dos antidepressivos analisados, os que apresentaram efeitos relevantes na interação com a varfarina foram, em ordem decrescente: paroxetina, venlafaxina, fluoxetina e duloxetina.

The aim of the study was to analyze the distribution of the methylenetetrahydrofolate reductase (MTHFR), methionine synthase reductase (MTRR), and methionine synthase (MTR) polymorphisms in idiopathic infertile Brazilian men and fertile men. Case–control study comprising 133 idiopathic infertile Brazilian men with nonobstructive azoospermia ([NOA] n = 55) or severe oligozoospermia ([SO] n = 78) and 173 fertile men as controls. MTHFR C677T, A1298C, and G1793A; MTRR A66G; and MTR A2756G polymorphisms were studied by quantitative polymerase chain reaction (qPCR). The results were analyzed statistically and a P value

Many theories have been proposed to explain the development of endometriosis, and recently, autoimmune aetiology has been suggested. Besides, it is well known that endometriosis, especially the advanced disease, may impair fertility. B lymphocyte stimulator (BLyS) is a cytokine produced by macrophages and is necessary for normal B cell development. One of the most studied polymorphisms is the -817C/T in the promoter region of the gene. We aimed to assess the association between endometriosis-related infertility and idiopathic infertility and the BLyS -817C/T polymorphism in a Brazilian population. We performed a case–control study comprising 165 infertile women with endometriosis, 83 with idiopathic infertility and 145 fertile and assessed the association with BLys -817C/T polymorphism. BLyS -817C/T polymorphism was detected using TaqMan PCR. The results were analysed statistically, and a P-value

An aberrant immunologic mechanism has been suggested to be involved in the pathogenesis of endometriosis. Fc receptor–like 3 gene ( FCRL3 ) has been proposed as a novel autoimmune predisposing factor. The authors have hypothesized a possible relationship between endometriosis, infertility, and FCRL3 polymorphisms. This was a case-control study that included 170 women with endometriosis-related infertility, 91 women with idiopathic infertility, and 166 controls. Detection of FCRL3 polymorphisms (−169C/T, −110G/A, +358C/G and +1381 A/G) was performed using TaqMan PCR. The results were analyzed statistically and a p value Results Single-marker analysis revealed that FCRL3 −169C/T was significantly associated with endometriosis ( p = 0.004), regardless of the stage of the disease, p = 0.011 and p = 0.035, respectively to minimal/mild and to moderate/severe endometriosis. No association was found considering −110A/G, +358C/G, and +1381 A/G polymorphisms either for the endometriosis-related infertility group or the idiopathic infertility group. Haplotype analysis of four FCRL3 polymorphisms identified a haplotype GGGC associated with endometriosis ( p = 0.026). The haplotype AGAT was associated with protection against endometriosis ( p = 0.011) and infertility ( p = 0.041). The data from this study point to a possible association of the FCRL3 −169C/T polymorphisms with endometriosis, especially minimal/mild endometriosis, and the haplotype AGAT may be protective against the development of the disease, in Brazilian women. However, these findings clearly need to be replicated in an independent sample and in different populations.

Objective To evaluate FOXP3 polymorphisms (rs3761549, rs3761548, rs2232368, rs2232366, and rs2280883) in a group of infertile women with and without endometriosis and controls. Design Case control study. Setting Human Reproduction Outpatient Clinic of Faculdade de Medicina do ABC. Patient(s) The study groups were 177 infertile women with endometriosis, 71 women with idiopathic infertility, and 171 fertile women as controls. Intervention(s) The FOXP3 polymorphisms were identified by TaqMan polymerase chain reaction (PCR). The results were analyzed statistically. Main Outcome Measure(s) Genotype distribution, allele frequency, and haplotype analysis of the FOXP3 polymorphisms. Result(s) : Single-marker analysis revealed that FOXP3 rs3761549 was significantly associated with endometriosis. In the infertile group without endometriosis, single-marker analysis revealed statistical difference for rs2280883 and rs2232368 FOXP3 polymorphisms. No associations were found with rs3761548 and rs2232366 either for endometriosis-related infertility group or idiopathic infertility group. Haplotype analysis of five FOXP3 polymorphisms identified a haplotype CTTGA associated with endometriosis and ACTAG associated with idiopathic infertility. Conclusion(s) : This is the first study to report an association between FOXP3 polymorphisms and endometriosis and/or infertility. These findings require replication in other populations but suggest that the FOXP3 polymorphisms can be associated with risk of idiopathic infertility (rs2280883 and rs2232368) and endometriosis (rs3761549) in Brazilian women.

An aberrant immunological mechanism is thought to be involved in the pathogenesis of endometriosis. The present study aimed to determine whether there is a relationship between endometriosis and/or infertility and the FCRL3 C-169T polymorphism. This case-control study included 167 infertile women with endometriosis, 60 women with idiopathic infertility and 167 fertile women. Detection of the FCRL3 C-169T polymorphism was performed using TaqMan PCR. A significant difference in the genotype and allele frequencies of the FCRL3 C-169T polymorphism between endometriosis-related infertility ( p =0.003 and p =0.001) and idiopathic infertility ( p =0.027 and p =0.0185) versus controls was demonstrated. In conclusion, the results suggest that the FCRL 3 C-169T polymorphism may play an important role in the pathogenesis of endometriosis and/or infertility.

Estrogens are important factors in the development of endometriosis, and can induce cell proliferation and stimulate cell division. COMT constitutes a crucial element in estrogen metabolism and has been suggested to be involved in the development of endometriosis. This study had the objective of to determine whether the presence of COMT val/met polymorphism (rs4680) increases the risk to endometriosis in infertile patients. A case-control study that included 198 infertile women with endometriosis, 71 infertile women without endometriosis, and 168 fertile women as control group of the Faculdade de Medicina do ABC. COMT (val/met) genotypes were identified by real time PCR (genotyping TaqMan assay) and the results were analyzed statistically by χ² test. The data showed no statistical difference in the distribution of COMT genotypes neither between infertile patients with endometriosis and control group (p = 0.567), regardless disease degree, nor between infertile patients without endometriosis and control group (p = 0.460). In conclusion, the COMT val/met polymorphism is not associated to endometriosis-related infertility in the Brazilian population evaluated. However, more studies in larger populations are necessary to confirm these results.

An aberrant immunologic mechanism has been suggested to be involved in the pathogenesis of endometriosis. Genetic alterations in the vitamin D receptor gene (VDR) may lead to important defects in gene activation that principally affect immune function. We have hypothesized a possible relationship between endometriosis and/or infertility and the VDR polymorphisms (ApaI, TaqI, FokI, and BmsI). The study was a case–control study including 132 women with endometriosis-related infertility, 62 women with idiopathic infertility, and 133 controls. VDR polymorphisms were studied by restriction fragment length polymorphism. We found relatively similar VDR polymorphism genotype frequencies in cases and controls. When patients with minimal/mild and moderate/severe endometriosis were studied separately, no difference was found. When we compared infertile groups with and without endometriosis there was no statistically significant difference. The data suggest that VDR polymorphisms did not play an important role in the pathogenesis of endometriosis and/or infertility in the Brazilian women studied.

Summary An autoimmune aetiology has been suggested for endometriosis mostly on the basis of an increased prevalence of autoimmune diseases in affected women. Cytotoxic T lymphocyte antigen (CTLA) 4 gene is recognized as a primary determinant for autoimmunity, since specific polymorphisms have been associated with predisposition to most autoimmune disorders. Thus, the objective of the study was to evaluate CTLA4 polymorphism (+49A/G) in a group of infertile women with and without endometriosis and controls. Case–control study comprising 244 infertile women (177 with endometriosis and 67 without endometriosis) and 172 fertile women as controls. CTLA4 polymorphism was identified by qPCR. The results were analysed statistically and a P-value

Citation Gomes FMCS, Bianco B, Teles JS, Christofolini DM, de Souza AMB, Guedes AD, Barbosa CP. PTPN22 C1858T polymorphismin women with endometriosis. Am J Reprod Immunol 2010; 63: 227–232 Problem Endometriosis has been suggested to be an autoimmune disease and recently, an allelic variation of the PTPN22 (C1858T) gene was revealed to be associated with the development of autoimmunity. The aim of the study was to determine the frequency of the PTPN22 (C1858T) polymorphism in Brazilian women with endometriosis as compared with controls. Method of study Case–control study included 140 women with endometriosis and a control group consisting of 180 healthy fertile women without a history of endometriosis and/or autoimmune diseases from the ABC School of Medicine. The PTPN22 (C1858T) polymorphism was studied by restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR). Results Genotypes CC, CT and TT of PTPN22 polymorphism presented frequencies of 67.9, 30.0 and 2.1% in the women with endometriosis (P = 0.008); 76.2, 19.0 and 4.8% in women with minimal/mild endometriosis (P = 0.173); 61.0, 39.0 and 0.0% in women with moderate/severe endometriosis (P ≤ 0.001) and 82.8, 16.1 and 1.1% in control group. Allele C and T were present in 82.9 and 17.1%; 85.7 and 14.3%; 80.5 and 19.5%; and 90.8 and 9.2% respectively, in women with endometriosis (P = 0.004), women with minimal/mild endometriosis (P = 0.148), women with moderate/severe endometriosis (P = 0.002) and control group. Conclusion The data suggest that in Brazilian women polymorphism PTPN22 (C1858T) may be an important genetic predisposing factor for endometriosis, especially, in advanced disease.

The objective of this study was to evaluate the disposition kinetics of enrofloxacin (ENR) in the plasma and its distribution in the muscle tissue of Nile tilapia (Oreochromis niloticus) after a single oral dose of 10 mg/kg body weight via medicated feed. The fish were kept at a temperature between 28 and 30 °C. The collection period was between 30 min and 120 h after administration of the drug. The samples were analyzed by high-performance liquid chromatography with a fluorescence detector (HPLC-FLD). The ENR was slowly absorbed and eliminated from the plasma (Cmax = 1.24 ± 0.37 μg/mL; Tmax = 8 h; T1/2Ke = 19.36 h). ENR was efficiently distributed in the muscle tissue and reached maximum values (2.17 ± 0.74 μg/g) after 8 h. Its metabolite, ciprofloxacin (CIP), was detected and quantified in the plasma (0.004 ± 0.005 μg/mL) and muscle (0.01 ± 0.011 μg/g) for up to 48 h. After oral administration, the mean concentration of ENR in the plasma was well above the minimum inhibitory concentrations (MIC50 ) for most bacteria already isolated from fish except for Streptococcus spp. This way the dose used in this study allowed for concentrations in the blood to treat the diseases of tilapia.

O lupus eritematoso discoide e uma desordem autoimune que geralmente afeta areas da pele expostas ao sol. A apresentacao de lesoes palpebrais na ausencia de outras anormalidades cutâneas e incomum, sendo o envolvimento da palpebra inferior prevalente em apenas 6% dos pacientes com lupus eritematoso cutâneo cronico. Relatamos o caso de uma paciente do sexo feminino de 40 anos, com hiperemia, madarose e ulceracao na palpebra inferior do olho esquerdo refrataria ao tratamento para blefarite. Inicialmente, outra lesao semelhante havia sido descrita na palpebra superior do olho direito. No entanto, as biopsias incisionais mostraram-se inconclusivas. Diante de uma lesao migratoria palpebral de caracteristicas semelhantes a primeira, a hipotese clinica de lupus eritematoso discoide foi aventada e o diagnostico confirmado por meio de revisao histopatologica. Uma forte suspeita clinica e o reconhecimento precoce podem evitar erros diagnosticos, complicacoes clinicas e tratamentos inapropriados, como descrito neste caso de lesao palpebral como manifestacao primaria do lupus eritematoso discoide.

Orientador: Felix Guillermo Reyes Reyes Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos Resumo: A enrofloxacina (ENR) é um antimicrobiano de exclusiva aplicação veterinária e possui como metabólito a ciprofloxacina (CIP), exclusiva de uso humano. No Brasil, a ENR não é permitida na aquicultura, porém, há evidências do seu uso nesse sistema de produção de alimentos. Por este motivo, em 2010, o MAPA (Ministério da Agricultura Pecuária e Abastecimento) através do Plano Nacional de Controle de Resíduos e Contaminantes em produtos de origem animal (PNCRC/Animal), a incluiu na lista dos antimicrobianos a serem monitorados em peixes de cultivo. Diante da intensificação na produção de tilápias no Brasil, da possível utilização indevida da ENR na piscicultura brasileira, da falta de informações terapêuticas com ENR para esta espécie e dos danos à saúde pública que isto pode gerar através do uso indiscriminado desse antimicrobiano, o presente trabalho teve o objetivo de realizar estudo farmacocinético de dose única ENR (10 mg kg-1 p.v) através de ração medicada (0.1% de ENR) em tilápia-do-Nilo (Oreochromis niloticus). A ENR (padrão analítico = 98%) foi dissolvida em solução alcoólica acidificada e adicionada a uma solução contendo o polímero, polivinilpirrolidona (PVP), e incorporada à ração comercial. O teor de ENR na ração foi quantificado (1.047 mg g-1) utilizando cromatografia líquida de alta eficiência, com detector de fluorescência (HPLC-FLD). Os peixes foram medicados através dessa ração e amostras de plasma e músculo foram coletadas nos seguintes tempos: 0.5, 1, 2, 4, 8, 12, 24, 48, 72, 96 e 120h após a administração. Foram desenvolvidos e validados métodos analíticos em HPLC-FLD para quantificação da ENR e CIP nas matrizes plasma e músculo. Os métodos analíticos apresentaram desempenho satisfatório na matriz plasma com limites de quantificação (LOQ) de 3 ng mL-1 obtidos para ambos analitos e 10.0 e 7.0 ng g-1, no músculo, para CIP e ENR, respectivamente. Em todos os métodos foram utilizadas para quantificação dos analitos, curvas analíticas construídas pela fortificação das matrizes. Quanto aos parâmetros farmacocinéticos obtidos da ENR, para plasma e músculo, a concentração máxima obtida foi de 1.241 ± 0.423 µg mL-1 e 2.166 ± 0.742 µg g-1, respectivamente, no tempo de 8h demonstrando absorção lenta. O alto volume de distribuição (Vd) no plasma (7.03L kg-1), os valores da relação entre concentração músculo/plasma (1.12 a 3.82) indicaram boa distribuição no tecido muscular. Foi possível detectar e quantificar o metabólito (CIP), porém a taxa de conversão foi baixa (1.3%). Por último, o T1/2Ke de 2.08 dias no músculo, indica eliminação lenta e longo período de carência para que o produto cárneo possa ser consumido. O uso de antimicrobianos sem dados toxicológicos e farmacológicos para a espécie a qual se destinam tem dentre várias consequências, o aumento do risco à exposição de resíduos com valores acima daqueles permitidos (LMRs) e aumento na incidência/prevalência de resistência bacteriana no animal e no ser humano Abstract: Enrofloxacin (ENR) is an exclusive antimicrobial veterinary application and has as metabolite ciprofloxacin (CIP), exclusive for human use. In Brazil, the ENR is not permitted in aquaculture, however, there is evidence of its use in this system of food production. For this reason, in 2010, MAPA (Ministry of Agriculture, Livestock and Supply) through the National Plan for Control of Residues and Contaminants in animal products (PNCRC/Animal), included in the list of antimicrobials to be monitored in fish cultivation. Given the intensification of tilapia production in Brazil, the possible improper use of ENR in Brazilian farming, the lack of information with therapeutic ENR for this species and harm to public health this could generate through the indiscriminate use of this antimicrobial, the aim of this work was to realize a single-dose pharmacokinetic study with ENR (10 mg k-1 bw) through medicated feed (0.1% ENR) in the Nile tilapia (Oreochromis niloticus). ENR (analytical standard = 98 %) was dissolved in acidified alcohol solution and added to a solution containing the polymer, polyvinylpyrrolidone (PVP ) and incorporated into commercial feed. The content of ENR in feed medicated was quantified (1.047 mg g-1) using high performance liquid chromatography with fluorescence detection (HPLC-FLD). The fish were medicated and its plasma and muscle samples were collected at the following times: 0.5, 1, 2, 4, 8, 12, 24, 48, 72, 96 and 120h after administration. Analytical methods were developed and validated for HPLC-FLD for quantification of ENR and CIP in plasma and muscle. Analytical methods showed satisfactory performance in plasma with limits of quantification (LOQ ) of 3 ng mL-1obtained for both analytes, and in muscle,7.0 and 10.0 ng g-1 for CIP and ENR, respectively. In all methods were used for quantification of analytes, standard curves constructed by fortification of matrices. About the pharmacokinetic parameters obtained from the ENR for plasma and muscle, the maximal concentration of ENR was 1.241 ± 0.423 mg ml-1 for plasma and 2.166 ± 0.742 mg g- 1 for muscle, after 8h showing slow absorption. The high volume of distribution (Vd) in plasma (7.03 L kg- 1), the values of the ratios between muscle concentration/plasma (1.12 to 3.82 ) indicated good distribution in muscle tissue. It was possible to detect and quantify the metabolites (CIP), but the conversion rate was low (1.3%). Finally, the T1/2Ke 2.08 days in muscle indicates slow elimination and suggested long period of time for the meat product to be consumed. The use of antimicrobial agents without pharmacological and toxicological data for the species which it is applied have several consequences, like increasing the risk of exposure of residues above those permitted ( MRLs) and increased incidence/prevalence of bacterial resistance in animals and humans Doutorado Ciência de Alimentos Doutora em Ciência de Alimentos

Objective To consider a possible cumulative effect of two genetic polymorphisms ( FOXP3 C-2383T/rs3761549 and FCRL3 C-169T/rs7528684) that were previously shown to be associated with endometriosis. Design Genetic association study. Setting Human reproduction outpatient clinic of Faculdade de Medicina do ABC. Patient(s) One hundred eighty-eight infertile women with endometriosis and 169 controls. Intervention(s) Detection of polymorphisms FOXP3 (C-2383T/rs3761549) and FCRL3 (C-169T/rs7528684) by TaqMan real-time polymerase chain reaction. The results were analyzed statistically. Main Outcome Measure(s) Genotype distribution, allele frequency, and combination analysis of the FOXP3 and FCRL3 polymorphisms. Result(s) Single-marker analysis revealed a significant association of FOXP3 C-2383T and FCRL3 C-169T, independently, with endometriosis-related infertility, regardless of the stage of the disease. Considering the combined genotypes of FCRL3 and FOXP3 polymorphisms, a positive association was found between genotypes FCRL3 TT/ FOXP3 CT, FCRL3 CT/ FOXP3 CT, and FCRL3 CC/ FOXP3 CT and the risk of endometriosis development. Moreover, a progression of the disease risk was observed according to the presence of one or two copies of risk allele FCRL3 C and only one copy of risk allele FOXP3 T (odds ratio [OR] = 2.14, OR=3.25, and OR=6.0, respectively, for genotypes FCRL3 TT/ FOXP3 CT, FCRL3 CT/ FOXP3 CT, and FCRL3 CC/ FOXP3 CT). Conclusion(s) Our findings support a possible gene-gene interaction leading to a cumulative effect on endometriosis development.

OBJECTIVE: To determine the frequency of the estrogen receptor b gene (ERβ) +1730 G/A polymorphism in infertile women with and without endometriosis and controls. SUBJECTS AND METHODS: Case-control study that included 136 women with endometriosis, 69 women without endometriosis and 209 fertile women as controls. The ERβ gene + 1730 G/A polymorphism was identified by RFLP-PCR (Restriction Fragment Length Polymorphism - Polymerase Chain Reaction). RESULTS: Genotypes GG, GA and AA of the ERβ gene presented frequencies of 60.3%, 38.2% and 1.5%, respectively, in the women with endometriosis (p < 0.0022). Of the infertile women without endometriosis, 63.8% presented the normal homozygous genotype GG, 30.4% the GA heterozygous genotype, and 5.8% the homozygous mutated genotype AA (p < 0.0275). In the control group, 77.5% presented the normal homozygous genotype GG, 21.1% the heterozygous genotype GA, and 1.4% the homozygous mutated genotype AA. CONCLUSION: The data suggest that the estrogen receptor β gene (ERβ) +1730 G/A polymorphism can be associated with risk of infertility and endometriosis-associated infertility. OBJETIVO: Determinar a frequência do polimorfismo +1730 G/A do gene do receptor beta de estrógeno (ERβ) em mulheres inférteis com e sem endometriose e controles. SUJEITOS E MÉTODOS: Estudo caso-controle que incluiu 136 mulheres com endometriose, 69 mulheres sem endometriose e 209 mulheres férteis como controles. O polimorfismo ERβ + 1730 G/A foi identificado por RFLP-PCR (Restriction Fragment Length Polymorphism - Polymerase Chain Reaction). RESULTADOS: Os genótipos GG, GA e AA do polimorfismo ERβ + 1730 G/A apresentaram frequência de 60,3%, 38,2% e 1,5%, respectivamente, nas mulheres com endometriose (p = 0,0022). Das mulheres inférteis sem endometriose, 63,8% apresentaram o genótipo homozigoto normal GG, 30,4% o genótipo heterozigoto GA e 5,8% o genótipo homozigoto mutado AA (p = 0,0275). No grupo controle, os genótipos GG, GA e AA apresentaram frequência de 77,5%, 21,1% e 1,4%. CONCLUSÃO: Os dados sugerem que o polimorfismo ERβ +1730G/ pode estar associado ao risco de infertilidade e infertilidade associada à endometriose.

Endometriosis has been suggested to be an autoimmune disease and recently, an allelic variation of the PTPN22 (C1858T) gene was revealed to be associated with the development of autoimmunity. The aim of the study was to determine the frequency of the PTPN22 (C1858T) polymorphism in Brazilian women with endometriosis as compared with controls.Case-control study included 140 women with endometriosis and a control group consisting of 180 healthy fertile women without a history of endometriosis and/or autoimmune diseases from the ABC School of Medicine. The PTPN22 (C1858T) polymorphism was studied by restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR).Genotypes CC, CT and TT of PTPN22 polymorphism presented frequencies of 67.9, 30.0 and 2.1% in the women with endometriosis (P = 0.008); 76.2, 19.0 and 4.8% in women with minimal/mild endometriosis (P = 0.173); 61.0, 39.0 and 0.0% in women with moderate/severe endometriosis (Por = 0.001) and 82.8, 16.1 and 1.1% in control group. Allele C and T were present in 82.9 and 17.1%; 85.7 and 14.3%; 80.5 and 19.5%; and 90.8 and 9.2% respectively, in women with endometriosis (P = 0.004), women with minimal/mild endometriosis (P = 0.148), women with moderate/severe endometriosis (P = 0.002) and control group.The data suggest that in Brazilian women polymorphism PTPN22 (C1858T) may be an important genetic predisposing factor for endometriosis, especially, in advanced disease.

Orientador: Roberto Herminio Moretti Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos Resumo: O uso de extrato de malte concentrado como matéria-prima cervejeira até hoje foi pouco estudado. É sabido que produzir cervejas claras e sem resíduo adocicado, é um desafio para este ingrediente. No entanto, cervejas produzidas a partir de extrato de malte concentrado, necessitam de menores investimentos quando se comparado ao método tradicional pois, economiza em mão-de-obra, espaço e energia. Pensando nisso, o trabalho teve como objetivo, produzir diferentes mostos concentrados lupulados formulados conforme planejamento experimental aplicado (DCCR), caracterizar as cervejas produzidas com estes mostos. Estudar o uso de xarope de alta maltose com a finalidade de redução da cor e proteína no produto final e verificar durante a fervura, interações ocorridas entre malte/adjunto, lúpulo e tempo quanto à cor, amargor, polifenóis e proteínas. Verificou-se que mostos produzidos com extrato de malte concentrado possuem fermentações mais lentas. Os produtos obtidos tiveram características físico-químicas semelhantes à cerveja tipo Pilsen. Durante a fervura, o tempo foi mais influente para a produção de cervejas mais escuras. Apesar disso, o uso de xarope de alta maltose como substituição parcial dos açúcares fermentescíveis do malte, na redução da cor, foi totalmente eficaz, como também para o aumento na luminosidade e menores quantidades de proteínas. Por fim, para promover mais eficiente complexação entre proteínas-polifenóis, seriam necessárias quantidades maiores de lúpulo. Assim sendo, mostos concentrados feitos a partir de extrato de malte concentrado possuem grandes potencialidades e levam a crer que acrescentam mais vantagens do que desvantagens para a produção de cerveja Abstract: The use of extract malt concentrated as beer raw material until today little was studied. It is known that to produce clear beers and without sweet residue, it is a challenge for this ingredient. However, beers produced from extract malt concentrated, need lesser investments when if compared with the traditional method, therefore it is saved in man power, space and energy. Thinking about this, the work had as objective, to produce differents worts hopped concentrated formulated as experimental design applied, to characterize the beers produced with these worts, to study the use of boiled must of high maltose with the purpose of reduction of the color and protein and to verify, during the boil, occurred interactions between malt/adjunct, hop and time how much to the color, bitter taste, polyphenol and proteins. It was verified that worts produced with extract of malt concentrated have slower fermentations, the gotten products had characteristics similar to the beer Pilsen type. During the boil, the time was more important for production of darker beers. Despite this, the use of high maltose syrup as partial substitution of the fermentable sugars of malt, in the reduction of the color was total efficient, as also for the increase of the luminosity and minors amounts of protein. Finally, to promote more efficient complex between proteins-polyphenols, they would be necessary more of hop. Thus being, made intent worts from extract of malt concentrated have great potentialities and lead to believe that they add more advantages of that disadvantages for beer production Mestrado Mestre em Tecnologia de Alimentos

A method for the identification and quantification of macrolides (erythromycin, josamycin, tilmicosin, tylosin, spiramycin and neospiramycin) in tilapia fillets by liquid chromatography coupled to quadrupole time of flight mass (LC–QToF) spectrometry is presented. Sample preparation was quite simple and low cost: proteins were precipitated and the analytes were extracted with ethanol, extract was defatted with hexane and concentrated by solvent evaporation. The matrix effect was statistically demonstrated during method validation, in which matrix-matched calibration was applied. Matrix effect mechanism was clarified thanks to the capability of QToF mass spectrometer of generating full scan spectra with accurate mass measurement. The limits of quantification were at least 45% lower than the maximum residue limits. The method was able to identify the studied macrolides with relative m/z errors lower than 2.5 ppm and to monitor two fragment ions per analyte, which is in accordance with the European Community recommendations for the analysis of contaminants in foods. Samples from the retail market of Sao Paulo State, Brazil, were analyzed by the developed method and none of them presented positive results for the macrolides studied.

Anthropogenic actions, including deforestation, disorganized urbanization, and globalization, contribute to emergence and reemergence of arboviruses worldwide, where Flavivirus is the most prevalent, and its continuous monitoring can help in preventive control strategies. Thus, the aim of this study was to detect flavivirus RNA in single hematophagous insects, which are used as sentinels. Total RNA was extracted from six Aedes aegypti stored since 2003 and from 100 Culicidae and collected through CDC trap in a public park of a Brazilian Northwest city of São Paulo State. Flavivirus was detected through RT/PCR targeting 230–250 bp of the RNA polymerase coding sequence (NS5). PCR amplicons were sequenced by Sanger method, used in comparative analysis over Basic Local Alignment Search Tool (BLAST) in GenBank, and subjected to Neighbor-Joining phylogenetic analyses. Efficiency of Flavivirus diagnosis was confirmed by detection of Dengue virus serotype 2 in Ae. aegypti. From the 100 collected insects, 19 were positive for Culex flavivirus (CxFV). NS5 partial sequence phylogenetic analysis clustered all CxFV in one branch separated from vertebrate flaviviruses, being applicable to the identification of Flavivirus species. The dipteran RNA extraction methodology described in this work supports detection of flaviviruses in single insects maintained in 80% ethanol, which can be used to constant arbovirus surveillance.