Bee Venom Toxic Effect on MDA-MB-231 Breast Cancer Cells and Caenorhabditis Elegans.

INTRODUCTION: Bee venom has therapeutics and pharmacological properties. Further toxicological studies on animal models are necessary due to the severe allergic reactions caused by this product. METHOD: Here, Caenorhabditis elegans was used as an in vivo toxicity model, while breast cancer cells were used to evaluate the pharmacological benefits. The bee venom utilized in this research was collected from Apis mellifera species found in Northeast Brazil. The cytotoxicity caused by bee venom was measured by MTT assay on MDA-MB-231 and J774 A.1 cells during 24 - 72 hours of exposure. C. elegans at the L4 larval stage were exposed for three hours to M9 buffer or bee venom. Survival, behavioral parameters, reproduction, DAF-16 transcription factor translocation, the expression of superoxide dismutase (SOD), and metabolomics were analyzed. Bee venom suppressed the growth of MDA-MB-231 cancer cells and exhibited cytotoxic effects on macrophages. Also, decreased C. elegans survival impacted its behaviors by decreasing C. elegans feeding behavior, movement, and reproduction. RESULTS: Bee venom did not increase the expression of SOD-3, but it enhanced DAF-16 translocation from the cytoplasm to the nucleus. C. elegans metabolites differed after bee venom exposure, primarily related to aminoacyl- tRNA biosynthesis, glycine, serine and threonine metabolism, and sphingolipid and purine metabolic pathways. Our findings indicate that exposure to bee venom resulted in harmful effects on the cells and animal models examined. CONCLUSION: Thus, due to its potential toxic effect and induction of allergic reactions, using bee venom as a therapeutic approach has been limited. The development of controlled-release drug strategies to improve this natural product's efficacy and safety should be intensified.

Narrowing the Relationship between Human CCR5 Gene Polymorphisms and Chagas Disease: Systematic Review and Meta-Analysis.

Our aim was to carry out a qualitative and quantitative synthesis of the influence of CCR5 genetic variants on Chagas disease (CD) through a systematic review. A total of 1197 articles were analyzed, and eleven were included in the review. A meta-analysis was conducted along with principal component analyses (PCAs). The polymorphisms found were analyzed using the SNP2TFBS tool to identify possible variants that influence the interaction with gene binding sites. Eleven studied variants were identified: rs2856758, rs2734648, rs1799987, rs1799988, rs41469351, rs1800023, rs1800024, Δ32/rs333, rs3176763, rs3087253 and rs11575815. The studies analyzed were published between 2001 and 2019, conducted in Argentina, Brazil, Spain, Colombia and Venezuela, and included Argentine, Brazilian, Colombian, Peruvian and Venezuelan patients. Eight polymorphisms were subjected to the meta-analysis, of which six were associated with the development of the cardiac form of CD: rs1799987-G/G and G/A in the dominance model and G/G in the recessiveness model; rs2856758-A/G in the codominance model; rs2734648-T/T and T/G in the dominance model; rs1799988-T/T in both the codominance and recessiveness models; rs1800023-G allele and the G/G genotype in the codominance and recessiveness models, and the G/G and G/A genotypes in the dominance model; and rs1800024-T allele. The PCA analyses were able to indicate the relationships between the alleles and the genotypes of the polymorphisms. The SNP2TFBS tool identified rs1800023 as an influencer of the Spi1 transcription factor (p < 0.05). A correlation was established between the alleles associated with the cardiac form of CD in this review, members of the C haplotype of the gene (HHC-TGTG), and the cardiac form of CD.

Perfil ginecológico e obstétrico de usuárias que realizam o exame papanicolau / Gynecological and obstetric profile of users who perform the pap smear / Perfil ginecológico y obstétrico de las usuarias que realizan la prueba de papanicolaou

Objetivo: Identificar o perfil ginecológico e obstétrico de mulheres que realizam o exame Papanicolau em uma população do Nordeste, Brasil. Métodos: Estudo descritivo com abordagem quantitativa, realizado em unidades básicas de saúde entre os anos de 2014 e 2018. Resultados: Do total de 724 mulheres atendidas, 33,7% (n=244) tinham idade ≥48 anos e 64,2% (n=465) se autodeclararam pardas. A faixa etária mais prevalente da menarca foi de 13 a 15 anos, com 46,1% (n=334), e a da coitarca foi de 16 a 18 anos, com 39,1% (n=283). Os dados ainda evidenciaram que 58,6% (n=424) tiveram de 1 a 5 gestações e 32% (n=232) relataram a primeira gestação entre 18 e 21 anos. Conclusão: Conhecer o perfil desta população é de suma importância para identificação das principais vulnerabilidades do grupo, de modo que as estratégias de promoção, proteção e recuperação da saúde sejam condizentes com a realidade vivenciada por essas mulheres (AU).

Objective: To identify the gynecological and obstetrical profile of women who undergo the Pap smear in a population in Northeast Brazil. Methods: Descriptive study with a quantitative approach, carried out in basic health units between 2014 and 2018. Results: Of the total of 724 women assisted, 33.7% (n=244) were aged ≥48 years and 64.2% (n=465) self-declared brown. The most prevalent age group at menarche was 13 to 15 years old, with 46.1% (n=334), and that of coitarche was 16 to 18 years old, with 39.1% (n=283). The data also showed that 58.6% (n=424) had 1 to 5 pregnancies and 32% (n=232) reported their first pregnancy between 18 and 21 years old. Conclusion: Knowing the profile of this population is of paramount importance for identifying the main vulnerabilities of the group, so that health promotion, protection and recovery strategies are consistent with the reality experienced by these women (AU).

Objetivo: Identificar el perfil ginecoobstétrico de mujeres que se realizan el Papanicolaou en una población del Nordeste de Brasil. Métodos: Estudio descriptivo con abordaje cuantitativo, realizado en unidades básicas de salud entre 2014 y 2018. Resultados: Del total de 724 mujeres atendidas, 33,7% (n=244) tenían edad ≥48 años y 64,2% (n=465) marrón autodeclarado. El grupo etario más prevalente de menarquia fue de 13 a 15 años, con 46,1% (n=334), y el de coitarquia fue de 16 a 18 años, con 39,1% (n=283). Los datos también mostraron que el 58,6% (n=424) tuvo de 1 a 5 embarazos y el 32% (n=232) reportó su primer embarazo entre los 18 y los 21 años. Conclusión: Conocer el perfil de esta población es de suma importancia para identificar las principales vulnerabilidades del grupo, para que las estrategias de promoción, protección y recuperación de la salud sean acordes a la realidad que viven estas mujeres (AU).

Human papillomavirus infection and risk factors to cervical cancer in asymptomatic women in the region of the Northeast of Brazil / Infecção pelo papillomavirus humano e fatores de risco para o cancer cervical em mulheres assintomáticas em uma região do Nordeste do Brasil

Objective: This study sought to identify the presence of HPV infection and the risk factors related to susceptibility to cervical cancer in asymptomatic women. Methods: It is a cross-sectional study with 428 users attended Basic Health Units, in Arapiraca, Alagoas, Brazil. Sociodemographic, behavioral variables, and cytopathological reports were collected. Molecular detection of the HPV virus was performed by Nested-PCR. Statistical analysis was conducted with SPSS version 22.0. Results: A total of 428 women were studied, HPV DNA detected in 39.2% (n = 168), with a mean age of 41 years old. There was an association of HPV with use of oral contraceptives (p <0.016) and alcoholism (p <0.038). It was showed a higher frequency of positive HPV in women older than 25 years old (88.7%), up to 5 sexual partners (93.4%), up to 3 pregnancies (71.4%), and with the cytopathologic results within the limits of normality (61.9%). HPV was identified in 40.3% (104/258) of the women with results within the limits of normality. Conclusion: Our results suggest that the use of oral contraceptives and alcoholism may be considered as possible risk factors related to cervical oncogenesis. With this, it is necessary to propose interventions aimed at the health education of this population, actions of prevention, and early detection.

Objetivo: Este estudo buscou identificar a presença de infecção pelo HPV e os fatores de risco relacionados à suscetibilidade ao câncer do colo do útero em mulheres assintomáticas. Métodos: Trata-se de um estudo transversal com 428 usuários atendidos em Unidades Básicas de Saúde, em Arapiraca, Alagoas, Brasil. Foram coletados relatórios sociodemográficos, variáveis comportamentais e citopatológicos. A detecção molecular do vírus HPV foi realizada por Nested-PCR. A análise estatística foi realizada com SPSS versão 22.0. Resultados: Foram estudadas 428 mulheres, com DNA de HPV detectado em 39,2% (n =168), com média de idade de 41 anos. Houve associação do HPV com o uso de anticoncepcional oral (p<0,016) e alcoolismo (p <0,038). Foi evidenciada maior frequência de HPV positivo em mulheres maiores de 25 anos (88,7%), até cinco parceiros sexuais (93,4%), até três gestações (71,4%) e com resultados citopatológicos dentro dos limites da normalidade (61,9%). O HPV foi identificado em 40,3% (104/258) das mulheres com resultados dentro dos limites da normalidade. Conclusão: Nossos resultados sugerem que o uso de anticoncepcionais orais e o alcoolismo podem ser considerados como possíveis fatores de risco relacionados à oncogênese cervical. Com isso, é necessário propor intervenções voltadas para a educação em saúde dessa população, ações de prevenção e detecção precoce.

Gestation and breastfeeding in schistosomotic mice differentially alters the expression of histone deacetylases (HDACs) in adult offspring.

BACKGROUND: Breastfeeding or gestation in schistosomotic mothers can cause long-term alterations in the immune response of offspring. OBJECTIVES: Evaluate the expression of histone deacetylases (HDACs) (all classes), the production of cytokines by T and B lymphocytes and macrophages, and the frequency of CD4+CD25+FoxP3+-cells in adult offspring born and/or suckled by schistosomotic mothers. METHODS: We harvested splenocytes from offspring born to (BIM), suckled by (SIM), or born to/suckled by (BSIM) schistosomotic mothers and animals from noninfected mothers (Control) at seven-weeks old and cultured them with/without Concanavalin A. HDAC expression was evaluated by real-time quantitative polymerase chain reaction (qPCR), and cytokines and membrane markers were evaluated by fluorescence-activated cell sorting (FACS). FINDINGS: Compared to Control, BIM mice showed increased expression of HDAC9 and frequency of CD4+IL-10+-cells. The SIM group had increased expression of HDAC1, HDAC2, HDAC6, HDAC7, HDAC10, Sirt2, Sirt5, Sirt6, and Sirt7. The BSIM group only had increased HDAC10 expression. The SIM and BSIM groups exhibited decreased frequencies of CD4+IL-4+-cells and CD4+CD25+FoxP3+-cells, along with a higher frequency of CD14+IL-10+-cells and an increase in CD45R/B220+IL-10+-cells. The BSIM group also showed a high frequency of CD4+IL10+-cells. MAIN CONCLUSIONS: Breastfeeding induced the expression of HDACs from various classes involved in reducing inflammatory responses. However, gestation enhanced the expression of a single HDAC and breastfeeding or gestation appears to favour multiple IL-10-dependent pathways, but not cells with a regulatory phenotype.

Gestation and breastfeeding in schistosomotic mice differentially alters the expression of histone deacetylases (HDACs) in adult offspring

BACKGROUND Breastfeeding or gestation in schistosomotic mothers can cause long-term alterations in the immune response of offspring. OBJECTIVES Evaluate the expression of histone deacetylases (HDACs) (all classes), the production of cytokines by T and B lymphocytes and macrophages, and the frequency of CD4+CD25+FoxP3+-cells in adult offspring born and/or suckled by schistosomotic mothers. METHODS We harvested splenocytes from offspring born to (BIM), suckled by (SIM), or born to/suckled by (BSIM) schistosomotic mothers and animals from noninfected mothers (Control) at seven-weeks old and cultured them with/without Concanavalin A. HDAC expression was evaluated by real-time quantitative polymerase chain reaction (qPCR), and cytokines and membrane markers were evaluated by fluorescence-activated cell sorting (FACS). FINDINGS Compared to Control, BIM mice showed increased expression of HDAC9 and frequency of CD4+IL-10+-cells. The SIM group had increased expression of HDAC1, HDAC2, HDAC6, HDAC7, HDAC10, Sirt2, Sirt5, Sirt6, and Sirt7. The BSIM group only had increased HDAC10 expression. The SIM and BSIM groups exhibited decreased frequencies of CD4+IL-4+-cells and CD4+CD25+FoxP3+-cells, along with a higher frequency of CD14+IL-10+-cells and an increase in CD45R/B220+IL-10+-cells. The BSIM group also showed a high frequency of CD4+IL10+-cells. MAIN CONCLUSIONS Breastfeeding induced the expression of HDACs from various classes involved in reducing inflammatory responses. However, gestation enhanced the expression of a single HDAC and breastfeeding or gestation appears to favour multiple IL-10-dependent pathways, but not cells with a regulatory phenotype.

Putative biomarkers for cervical cancer: SNVs, methylation and expression profiles.

Cervical cancer is primarily caused by Human papillomavirus (HPV) infection, but other factors such as smoking habits, co-infections and genetic background, can also contribute to its development. Although this cancer is avoidable, it is the fourth most frequent type of cancer in females worldwide and can only be treated with chemotherapy and radical surgery. There is a need for biomarkers that will enable early diagnosis and targeted therapy for this type of cancer. Therefore, a systems biology pipeline was applied in order to identify potential biomarkers for cervical cancer, which show significant reports in three molecular aspects: DNA sequence variants, DNA methylation pattern and alterations in mRNA/protein expression levels. CDH1, CDKN2A, RB1 and TP53 genes were selected as putative biomarkers, being involved in metastasis, cell cycle regulation and tumour suppression. Other ten genes (CDH13, FHIT, PTEN, MLH1, TP73, CDKN1A, CACNA2D2, TERT, WIF1, APC) seemed to play a role in cervical cancer, but the lack of studies prevented their inclusion as possible biomarkers. Our results highlight the importance of these genes. However, further studies should be performed to elucidate the impact of DNA sequence variants and/or epigenetic deregulation and altered expression of these genes in cervical carcinogenesis and their potential as biomarkers for cervical cancer diagnosis and prognosis.

Collagenolytic enzymes produced by fungi: a systematic review

Abstract Specific proteases capable of degrading native triple helical or denatured collagen have been required for many years and have a large spectrum of applications. There are few complete reports that fully uncover production, characterization and purification of fungi collagenases. In this review, authors searched through four scientific on line data bases using the following keywords (collagenolytic OR collagenase) AND (fungi OR fungus OR fungal) AND (production OR synthesis OR synthesize) AND (characterization). Scientific criteria were adopted in this review to classify found articles by score (from 0 to 10). After exclusion criteria, 21 articles were selected. None obtained the maximum of 10 points defined by the methodology, which indicates a deficiency in studies dealing simultaneously with production, characterization and purification of collagenase by fungi. Among microorganisms studied the non-pathogenic fungi Penicillium aurantiogriseum and Rhizoctonia solani stood out in volumetric and specific collagenase activity. The only article found that made sequencing of a true collagenase showed 100% homology with several metalloproteinases fungi. A clear gap in literature about collagenase production by fungi was verified, which prevents further development in the area and increases the need for further studies, particularly full characterization of fungal collagenases with high specificity to collagen.

Infecção cervical por papilomavírus humano: genotipagem viral e fatores de risco para lesão intraepitelial de alto grau e câncer de colo do útero / Human papillomavirus cervical infection: viral genotyping and risk factors for high-grade squamous intraepithelial lesion and cervix cancer

OBJETIVO: analisar, em mulheres com HPV em colo do útero, as características da infecção viral e os fatores de risco para lesão intraepitelial de alto grau e carcinoma cervical. MÉTODOS: realizou-se um estudo caso-controle com mulheres com HPV em colo do útero atendidas em serviço de Ginecologia de referência vinculado ao SUS, em Recife, Nordeste do Brasil. No grupo de casos (72 mulheres com lesão intraepitelial de alto grau ou carcinoma cervical) e de controles (176 mulheres com colpocitologia normal ou com alterações benignas), foram pesquisados seis genótipos virais (HPV 16, 18, 31, 33, 6 e 11) em material da ecto- e endocérvice com primers MY09/MY11. As variáveis independentes foram hierarquizadas em três níveis de determinação: distal (sociodemográficas), intermediário (comportamentais) e proximal (realização anterior de colpocitologia). A homogeneidade das proporções foi testada (χ2). Obtiveram-se ORs não ajustadas e, na modelagem final, realizou-se regressão logística hierarquizada com o ajuste do efeito de cada variável sobre o desfecho pelas variáveis do mesmo nível e de níveis anteriores de causalidade. RESULTADOS: em 76,6 por cento das 248 mulheres participantes do estudo, o genótipo viral da infecção cervical foi identificado. Predominaram genótipos de alto risco oncogênico (83,4 por cento nos casos e 67,1 por cento nos controles), principalmente HPV 16 e 31. Foram identificados como fatores de risco (a) distais: residir em zona rural (OR=2,7; IC95 por cento: 1,1-6,2), menos de três anos de estudo (OR=3,9; IC95 por cento: 2,0-7,5) e renda familiar inferior a dois salários mínimos (OR=3,3; IC95 por cento: 1,0-10,5); (b) intermediário: número de gestações igual ou superior a quatro (OR=2,0; IC95 por cento: 1,0-3,7); (c) proximal: ausência de colpocitologia anterior (OR=9,7; IC95 por cento: 2,4-38,2). CONCLUSÕES: em mulheres usuárias do SUS do Nordeste do Brasil predominam os genótipos virais 16 e 31 em infecções cervicais ...

PURPOSE: to analyze the characteristics of viral infection and the risk factors for high-grade squamous intraepithelial lesion and cervical carcinoma in women with cervical HPV infection. METHODS: a case-control study was conducted on women with cervical HPV at a Gynecology reference service enrolled at the Public Health System, located in Recife, Northeastern Brazil. The groups of cases (72 women with high-grade squamous intraepithelial lesion or cervical cancer) and controls (176 women with normal Pap smear or benign alterations) were investigated for six viral genotypes (HPV 16, 18, 31, 33, 6, 11) in ecto- and endocervical material using MY09/MY11 primers. The independent variables were ranked in three levels of determination: distal (sociodemographic), intermediate (behavioral) and proximal (previous Pap smear). The homogeneity of proportions was tested (χ2), unadjusted Odds Ratios (OR) were obtained and hierarchical logistic regression was applied to the final model, with adjustment of the effect of each variable to the outcome based on the variables in the same and previous levels of causality. RESULTS: the viral genotype of cervical infection was identified in 76.6 percent of the 248 women participating in the study. High-risk HPV genotypes (83.4 percent of cases and 67.1 percent of controls) were predominant, especially HPV 16 and 31. The distal risk factors identified were: living in a rural area (OR=2.71, 95 percentCI: 1.18-6.23), less than three years of study (OR=3.97, 95 percentCI: 2.09-7.54) and family income below two minimum wages (OR=3.30, 95 percentCI: 1.04-10.51); intermediate: four or more pregnancies (OR=2.00, 95 percentCI: 1.06-3.76); and proximal: absence of a previous Pap smear (OR=9.74, 95 percentCI: 2.48-38.28). CONCLUSIONS: genotypes 16 and 31 of cervical HPV infection are predominant among women assisted by the Public Health System in Northeastern Brazil. Socioeconomic and reproductive factors, as well ...

Avaliação microbiológica da água de consumo empregada em unidades de alimentação / Microbiological evaluation of drinking water used in feeding units

A água de consumo humano é o principal veículo de transmissão de patógenos capazes de causar infecções gastrointestinais, sendo vital o seu controle microbiológico. O presente trabalho teve a finalidade de avaliar a qualidade bacteriológica da água para o consumo humano, quanto à presença de coliformes totais e termotolerantes, em diferentes unidades de alimentação na cidade do Recife (PE), conforme os padrões de potabilidade estabelecidos pela Portaria nº 518 de 2004 do Ministério da Saúde/ANVISA. Foram coletadas quarenta amostras de água em unidades de alimentação, localizadas no entorno da Universidade Federal de Pernambuco e da Universidade Federal Rural de Pernambuco. Foram realizados os testes bacteriológicos para pesquisa de coliformes totais e termotolerantes, conforme o protocolo do APHA (1995). Das amostras analisadas, 62,5 por cento apresentaram coliformes totais e 42,5 por cento, coliformes termotolerantes. De acordo com os resultados obtidos, 62,5 por cento das amostras não estão em conformidade com a legislação vigente.

The water for human consumption is the main vehicle for the transmission of pathogens that are capable of causing enteric parasitic diseases; therefore, its microbiological control is vital. This work had the purpose of evaluating the bacteriological quality of drinking water, in terms of the presence of total and thermotolerant coliforms, in different Feeding Units in the city of Recife, Pernambuco State, according to the potableness standards established by Ordinance No. 518 of 2004 of the Brazilian Ministry of Health/ANVISA. Forty water samples were collected in the Feeding Units located near the UFPE and UFRPE. The bacteriological tests for research on total and thermotolerant coliforms were carried out in accordance with the protocol of APHA (1995). Of the analyzed samples, 62.5 percent presented total coliforms and 42.5 percent thermotolerant coliforms. According to the results obtained, 62.5 percent of the samples are not in compliance with the law in force

Perfil de proteases de lesões cutâneas experimentais em camundongos tratadas com a lectina isolada das sementes de Canavalia brasiliensis / Proteases profile of skin wounds treated with lectin from Canavalia brasiliensis seeds

O objetivo deste estudo foi determinar o perfil de proteases em lesões cutâneas experimentais tratadas com a lectina isolada das sementes da Canavalia brasiliensis (ConBr) livre e conjugada com o seu açúcar específico. Lesões cirúrgicas foram produzidas assepticamente na região dorsal de camundongos (n=120), divididos de acordo com o tratamento empregado: Grupo NaCl (NaCl 150mM), Grupo manose (manose 100mM), Grupo ConBr (ConBr 100µg mL-1) e Grupo ConBr/manose (solução contendo ConBr 100µg mL-1 preparada em manose 100mM). Amostras da área lesada foram coletadas para determinação do perfil de proteases e atividade colagenolítica no 2°, no 7° e no 12° dia de pós-operatório. O perfil das proteínas realizado através de eletroforese SDS-PAGE demonstrou a presença de proteínas com massa molecular de 67kDa em todos os grupos. O Grupo ConBr/manose apresentou a maior atividade colagenolítica no 12° dia de pós-operatório. A lectina isolada das sementes da Canavalia brasiliensis influenciou a expressão de proteases com atividade colagenolítica podendo assim interferir no processo cicatricial das lesões cutâneas em camundongos.

The objective of the present study was determining the proteases profile of cutaneous healings treated with free and conjugated lectin of Canavalia brasiliensis (ConBr) and their specific sugar. An aseptic wound was produced in the thoracic area of the mice (n=120), divided according to the employed treatment: NaCl Group (150mM NaCl), manose Group (100mM manose), ConBr Group (100µg mL-1 ConBr) and ConBr/manose Group (solution containing 100µg mL-1 ConBr prepared in 100mM manose). Samples of the injured area were collected for determination of proteases profile and collagenolytic activity on 2nd, 7th e 12th days after the surgery. Electrophoresis SDS-PAGE demonstrated proteins with molecular mass of 67kDa in all groups. Group IV presented the highest collagenolytic activity on the 12th day post surgery. ConBr lectin influenced proteases expression with collagenolytic activity thus being able to intervene on skin wound healing in mice.

Proteínas do plasma seminal de caprinos relacionadas com o índice pluviométrico e a qualidade do sêmen / Proteins of goat seminal plasma related with precipitation index and semen quality

O objetivo deste estudo foi identificar as proteínas do plasma seminal de caprinos da raça Alpina Americana criados na região Nordeste do Brasil que estão relacionadas ao índice pluviométrico e à qualidade do sêmen. O sêmen foi obtido pelo método de vagina artificial a partir de três reprodutores e foi avaliado quanto aos parâmetros macroscópicos e microscópicos. O perfil de proteínas do plasma seminal foi realizado por eletroforese bidimensional. Os parâmetros volume do sêmen, integridade do acrossoma e proteínas totais evidenciaram diferença significativa (P<0,05) entre os períodos de alto (1,7mL, 90,3% e 372μg mL-1,respectivamente) e baixo (1,2mL, 80,3% e 494μg mL-1, respectivamente) índice pluviométrico. Foram detectados, nos períodos de alto e baixo índice pluviométrico, 47 e 49 spots de proteínas com massa molecular relativa de 4 a 106kDa e de 15 a 97kDa e ponto isoelétrico de 3,00 a 8,96 e de 4,48 a 9,83, respectivamente. Apenas no período de alto índice pluviométrico foram observados os grupo de proteínas de 13kDa e 45kDa. Conclui-se que o sêmen de caprinos da raça Alpina Americana criados na região Nordeste do Brasil apresenta-se com melhor qualidade quando colhido no período de alto índice pluviométrico, o que pode ser atribuído à presença das proteínas de 13kDa e 45kDa.

The aim of this study was to identify proteins in seminal plasma of goats raised in the Northeast of Brazil relatedwith precipitation index and semen quality. Semen was obtained from three bucks and evaluated to the microscopic and macroscopic parameters. The profile of seminal plasma proteins was performed by analysis of two-dimensional electrophoresis. Volume, acrosome integrity and total proteins had significant difference (P<0.05) between the periods of high (1.7mL, 90.3% and 372g mL-1, respectively) and low (1.2mL, 80.3% and 494μgmL-1, respectively) precipitation index. It was detected during high and low precipitation index, 47 and 49 spots of proteins with molecular weight of 4 to 106kDa and 15 to 97kDa, and isoelectric point of 3.00 to 8.96, and 4.48 to 9.83, respectively. Only in the period of high precipitation index were observed groups of proteins with 13kDa and 45kDa. It can be concluded that semen of Alpine American goats raised in the Northeast of Brazil has best quality when obtained in the period of high precipitation index, which can be attributed to the presence of protein with 13kDa and 45kDa.

Jacaratia corumbensis O. Kuntze a new vegetable source for milk-clotting enzymes

The partial characterization and purification of milk clotting enzyme obtained from the (root latex) of Jacaratia corumbensis O. kuntze was studied, by fractional precipitation with ammonium sulphate and ion exchange chromatography. The ammonium sulphate precipitate showed five fractions (AS1- 0-20 percent; AS2 - 20-40 percent; AS3 - 40-60 percent; AS4 - 60-80 percent; AS5 - 80-100 percent) and among the fractions obtained, the 40-60 percent fraction (AS3) showed the highest milk clotting activity with a purification factor of 1.2 fold in relation to the crude extract. This fraction when applied on Mono Q column yielded two protein peaks (p1 and p2), but p1 pool showed the best milk-clotting activity. The optimal pH for the crude and partially purified extract was 6.5 and 7.0, respectively. The maximum milk-clotting activity was at 55ºC for the both crude and partially purified extracts. The enzyme was inhibited by iodoacetic acid which suggested that this enzyme was a cysteine protease, with molecular weight of 33 kDa.

A enzima coagulante de leite obtida de látex de raiz de Jacaratia corumbensis O. kuntze foi caracterizada parcialmente e purificada, por precipitação fracionária com sulfato de amônio e cromatografia de troca de íon. Foram utilizadas cinco frações de sulfato de amônio (AS1 - 0-20 por cento; AS2 - 20-40 por cento; AS3 - 40-60 por cento; AS4 - 60-80 por cento; AS5 - 80-100 por cento), a fração 40-60 por cento (AS3) mostrou alta atividade coagulante com um fator de purificação de 1,2 vezes em relação ao extrato bruto. Esta fração foi aplicada em coluna Mono Q obtendo dois picos de proteína (p1 e p2), o p1 mostrou melhor atividade coagulante. O pH ótimo para o extrato bruto e parcialmente purificado foi 6,5 e 7,0, respectivamente. A atividade coagulante foi atingida a 55ºC para ambos os extratos, bruto e parcialmente purificado. A enzima foi inibida por ácido iodoacético que sugere que esta enzima é uma cisteína protease, com peso molecular de 33 kDa.

Comparative performance of microbial lipases immobilized on magnetic polysiloxane polyvinyl alcohol particles

Microbial lipase from Mucor miehei and Candida rugosa were immobilized by covalent binding onto magnetized polysiloxane polyvinyl alcohol particles (POS-PVA). The resulting immobilized derivatives were evaluated in aqueous solution (olive oil hydrolysis) and organic solvent (butyl butyrate synthesis). Higher catalytic activities were found when the coupling procedure was made with M. miehei lipase. Immobilized M. miehei lipase also showed a better operational stability and a higher half-life than C. rugosa lipase after the successive batches of esterification. The performance of C. rugosa immobilized derivative was constrained by the low lipase loading used in the immobilizing step. Further information regarding the both immobilized derivatives was also obtained through chemical composition (FTIR).

Lipases microbianas de Mucor miehei e Candida rugosa foram imobilizadas por ligação covalente em partículas magnetizadas de polisiloxano-álcool polivinílico (POS-PVA). Os derivados imobilizados resultantes foram avaliados em solução aquosa (hidrólise de azeite de oliva) e em solvente orgânico (síntese de butirato de butila). As maiores atividades catalíticas foram encontradas quando o procedimento de ligação foi realizado com lipase de M. miehei. O derivado imobilizado de lipase de M. miehei também apresentou melhores resultados de estabilidade operacional e tempo de meia-vida do que o de lipase de C. rugosa, após sucessivas bateladas de esterificação. O desempenho do derivado imobilizado de C. rugosa foi restringido pelo baixo carregamento de lipase usado na etapa de imobilização. Informações adicionais a respeito de ambos derivados imobilizados também foram obtidas através da composição química (FTIR).

Salmonella spp., importante agente patogênico veiculado em alimentos / Salmonella spp., important pathogenic agent transmitted through foodstuffs

A ocorrência de doenças transmitidas por alimentos (DTAs) tem sido foco de discussões nos últimos anos, devido à preocupação mundial com estratégias que permitam seu controle e, conseqüentemente, garantam a colocação de produtos seguros no mercado consumidor. A Salmonella spp. é um dos microrganismos mais amplamente distribuídos na natureza, sendo o homem e os animais seus principais reservatórios naturais, com ocorrência de sorotipos regionais, reconhecidos como salmoneloses, e considerado como um dos principais agentes envolvidos em surtos de origem alimentar em países desenvolvidos. O aumento da incidência da salmonelose provocada por alimentos contaminados demonstra que, na atualidade, apesar dos avanços tecnológicos alcançados, este problema ainda ocorre mundialmente. As aves e bovinos são responsáveis pela maior disseminação desse agente patogênico. A ampla distribuição da Salmonella spp. entre os animais, a existência de portadores assintomáticos e sua permanência no ambiente e nos alimentos contribuem para que este microrganismo assuma um papel de grande relevância na saúde pública mundial e, portanto, programas permanentes de controle e erradicação devem ser adotados.

The occurrence of foodborne diseases has been a matter of discussion over the last years due to the worldwide concern with strategies for their control and for ensuring that safe food products reach the consumer. Salmonella spp. is among the most widespread microorganisms in nature, having man and animals as main natural reservoirs. With occurrence of regional serotypes causing salmonellosis, this pathogen is considered one of the main agents responsible for outbreaks of foodborne disease in the developing countries. The increasing incidence of salmonellosis caused by contaminated food has shown that, despite the recent technological improvements, this problem still occurs in all countries. Cattle and poultry are the main responsible for the transmission of this pathogenic agent. Due to its wide distribution in animals, the existence of asymptomatic carriers and its presence in foodstuff and in the environment Salmonella spp. represents a significant public health problem worldwide calling for permanent control programs and eradication strategies.

Qualidade bacteriológica do coxão bovino em mercados públicos da cidade do Recife, PE / Bacteriological quality of beef in public markets in the city of Recife, PE

Foram analisadas 45 amostras de coxão duro bovino, comercializados em dois mercados públicos, localizados na Região Metropolitana do Recife, tendo como finalidade avaliar a qualidade bacteriológica e obter informações sobre parâmetros higiênico-sanitários. Os resultados confirmaram a presença de coliformes totais e Escherichia coli em 100 por cento das amostras, Staphylococcus spp. em 73 por cento, Clostridium perfringens em 46,6 por cento, Streptococcus spp. em 20 por cento, e ausência para Salmonella spp. em 100 por cento das amostras. Pode-se concluir que o coxão duro bovino comercializado nos mercados públicos indicaram condições inadequadas de manipulação e de estocagem, apesar de atender às exigência da legislação vigente, uma vez que esta estabelece ausência de Salmonella spp. para amostra indicativa em carne bovina.

Purification of plasmid (pVaxLacZ) by hydrophobic interaction chromatography

O DNA plasmidial tem sua aplicação na terapia gênica, tendo que apresentar elevado grau de pureza. Um método de purificação foi descrito neste trabalho, que inclui precipitação com sulfato de amônio seguida da cromatografia de interação hidrofóbica, usando a resina phenyl sepharose 6 fast flow. Ocorreu aumento da pureza das amostras analisadas, em torno de 30% após a precipitação com sulfato de amônio. Houve significativa redução dos contaminantes após a eluição das amostras na cromatografia de interação. O maior rendimento obtido foi 53% e fator de purificação 3,1 vezes após a eluição na cromatografia de interação hidrofóbica. A vantagem deste método é a possibilidade de explorar o caráter hidrofóbico, constituído de material genético de fita simples, tais como RNA e DNA genômico e proteínas, que podem interferir na liberação deste material para possíveis aplicações em terapia gênica.