Emergence of clonal complex 5 (CC5) methicillin-resistant Staphylococcus aureus (MRSA) isolates susceptible to trimethoprim-sulfamethoxazole in a Brazilian hospital

In this study, genotyping techniques including staphylococcal chromosomal cassette mec (SCCmec) typing, pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and restriction-modification tests were used to compare the molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) isolates recovered at two times within a 10-year interval (1998 and 2008) from a tertiary Brazilian hospital. In addition, the antimicrobial susceptibility profiles were analyzed. All 48 MRSA isolates from 1998 and 85.7% from 2008 (48/56 isolates) displayed multidrug-resistance phenotypes and SCCmec III. All but one of the 13 representative SCCmec III isolates belonged to CC8 and had PFGE patterns similar to that of the BMB9393 strain (Brazilian epidemic clone of MRSA; BEC). In 2008, we found an increased susceptibility to rifampicin and chloramphenicol among the SCCmec III isolates. In addition, we detected the entrance of diverse international MRSA lineages susceptible to trimethoprim-sulfamethoxazole (SXT), almost all belonging to CC5. These non-SCCmec III isolates were related to the USA 300 (ST8-SCCmec IV; PFGE-type B), USA 800 (ST5-SCCmec IV; subtype D1), USA 100 (ST5-SCCmec II; subtype D2), and EMRSA-3/Cordobes (ST5-SCCmec I, type C) clones. To the best of our knowledge, this is the first report of the emergence of isolates genetically related to the EMRSA-3/Cordobes clone in southeast Brazil. In this regard, these isolates were the most common non-SCCmec III MRSA in our institution, accounting for 8.9% of all isolates recovered in 2008. Thus, despite the supremacy of BEC isolates in our country, significant changes may occur in local MRSA epidemiology, with possible consequences for the rationality of MRSA empiric therapy.

Emergence of clonal complex 5 (CC5) methicillin-resistant Staphylococcus aureus (MRSA) isolates susceptible to trimethoprim-sulfamethoxazole in a Brazilian hospital.

In this study, genotyping techniques including staphylococcal chromosomal cassette mec (SCCmec) typing, pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and restriction-modification tests were used to compare the molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) isolates recovered at two times within a 10-year interval (1998 and 2008) from a tertiary Brazilian hospital. In addition, the antimicrobial susceptibility profiles were analyzed. All 48 MRSA isolates from 1998 and 85.7% from 2008 (48/56 isolates) displayed multidrug-resistance phenotypes and SCCmec III. All but one of the 13 representative SCCmec III isolates belonged to CC8 and had PFGE patterns similar to that of the BMB9393 strain (Brazilian epidemic clone of MRSA; BEC). In 2008, we found an increased susceptibility to rifampicin and chloramphenicol among the SCCmec III isolates. In addition, we detected the entrance of diverse international MRSA lineages susceptible to trimethoprim-sulfamethoxazole (SXT), almost all belonging to CC5. These non-SCCmec III isolates were related to the USA 300 (ST8-SCCmec IV; PFGE-type B), USA 800 (ST5-SCCmec IV; subtype D1), USA 100 (ST5-SCCmec II; subtype D2), and EMRSA-3/Cordobes (ST5-SCCmec I, type C) clones. To the best of our knowledge, this is the first report of the emergence of isolates genetically related to the EMRSA-3/Cordobes clone in southeast Brazil. In this regard, these isolates were the most common non-SCCmec III MRSA in our institution, accounting for 8.9% of all isolates recovered in 2008. Thus, despite the supremacy of BEC isolates in our country, significant changes may occur in local MRSA epidemiology, with possible consequences for the rationality of MRSA empiric therapy.

Uncaria tomentosa for Reducing Side Effects Caused by Chemotherapy in CRC Patients: Clinical Trial.

To evaluate the effectiveness of Uncaria tomentosa in minimizing the side effects of chemotherapy and improving the antioxidant status of colorectal cancer (CRC) patients, a randomized clinical trial was conducted. Patients (43) undergoing adjuvant/palliative chemotherapy with 5-Fluorouracil/leucovorin + oxaliplatin (FOLFOX4) were split into two groups: the UT group received chemotherapy plus 300 mg of Uncaria tomentosa daily and the C group received only FOLFOX4 and served as a control. Blood samples were collected before each of the 6 cycles of chemotherapy, and hemograms, oxidative stress, enzymes antioxidants, immunologic parameters, and adverse events were analyzed. The use of 300 mg of Uncaria tomentosa daily during 6 cycles of FOLFOX4 did not change the analyzed parameters, and no toxic effects were observed.

Postobturation pain and associated factors in adolescent patients undergoing one- and two-visit root canal treatment.

OBJECTIVES: This prospective study evaluated the frequency and intensity of postobturation pain and associated factors in adolescents undergoing one- and two-visit root canal treatment. METHODS: 121 patients aged 11-18 years presenting with molars with pulp necrosis were assigned randomly into two treatment groups: one- and two-visit (including interappointment dressing with calcium hydroxide paste). The canals of all teeth were prepared using a preflaring (2/3 middle-cervical) and step-back (1/3 apical) preparation techniques and filled with laterally compacted gutta-percha and sealer. Postobturation pain was recorded on a visual analogue scale (VAS) of 0-5. Data were statistically analyzed using multivariate logistic regression. RESULTS: The frequencies of postobturation pain were 10.5% (6/57) in the one-visit group and 23.0% (14/61) in the two-visit group. There were no statistically significant differences between the groups (p=0.07). The intensity of the pain was similar in both groups, particularly flare-ups, with a prevalence of 1.75% in the one-visit group and 1.65% in the two-visit group. Postobturation pain was significantly associated with the presence of preoperative pain (p=0.04; OR=3.54; CI 95%=1.02-12.30) and a positive culture at the time of obturation (p=0.00; OR=9.43; CI 95%=2.93-30.35). CONCLUSIONS: Postobturation pain was more present in the two-visit group, but was not statistically significant. The intensity of postobturation pain was similar. Effective microbiological control and the presence of preoperative pain may influence the postobturation pain in adolescents.

Características seminais, corpóreas e anatômicas do aparelho reprodutor de reprodutores da raça canchim aos 14 e 48 meses de idade. / Anatomical, corporal and seminal characteristics of the reproductive system of canchim breed reproductors at ages from 14 to 48 months / Características seminales, corpóreas y anatómicas del aparato reproductor de reproductores de la raza canchim a los 14 y 48 meses de edad

Foram utilizados 20 touros da raça Canchim em inatividade sexual, divididos em grupo 1 (G1), constituído de 10 animais com 14 meses de idade e grupo 2 (G2), com 10 animais de 48 meses de idade. A proposta desse estudo foi investigar as características dos ejaculados e das mensurações anatômicas do trato reprodutivo da raça Canchim. Os parâmetros obtidos foram: peso médio de 445,5 e 706,02 kg; circunferência escrotal de 31,80 e 36,25 cm e índice de massa corpórea de 270,33 e 346,73 kg/m2 para o G1 e G2, respectivamente, com diferenças significativas (p < 0,05) entre eles. Realizaram-se quatro colheitas de sêmen por touro por meio de eletroejaculação, com intervalos de 14 dias, verificando-se diferença significativa (p < 0,05) para os defeitos espermáticos totais, entre a primeira (5,48) e a terceira colheitas (1,08), no G2. Os resultados sugerem que o desenvolvimento do aparelho reprodutivo e a espermatogênese estão presentes aos 14 meses de idade.

Twenty bulls of the Canchin breed in sexual inactivity were divided in group 1(G1) with ten animals at the age of 14 months old and group 2 (G2) with ten animals at the age of 48 months old. The purpose of this study was to investigate the characteristics of the ejaculators and anatomic measurements of reproductive tract in Canchim breed. The parameters obtained were the average weight from 445.5 to 706.02 kg, scrotal circumference from 31.8 to 36.25 cm and Corporal Mass Index from 270.33 to 346.73 kg/m² for G1 and G2 respectively with a significant difference (p<0.05). Four semen collections per bull were made with electro ejaculation with intervals of 14 days, and it showed significant difference (p<0.05) for the total spermatic flaws, between the first (5.48) and the third collect (1.08) in G2. These results suggest that the development of the reproductive tract and the spermatogenesis are present at the age of 14 months old.

Fueron utilizados 20 toros de la raza Canchim en inactividad sexual, divididos en grupos:Grupo 1 (G1) con 10 animales, con edades de 14 meses y el grupo 2 (G2) con 10 animales, con edades de 48 meses, teniendo como objetivo verificar la adaptación al clima tropical, por medio de las características de los eyaculados y medidas anatómicas del aparato reproductivo. Los parámetros obtenidos fueron peso medio de 445,5 y 706,02 kg; circunferencia escrotal de 31,80 y 36,25 cm e índice de masa corpórea de 270,33 y 346,73 kg/m² para el G1 y G2, respectivamente, con diferencias significativas (p<0,05). Fueron realizadas cuatro colectas de semen por toro, con intervalo de 14 días, por medio de electroeyaculación verificando diferencias significativas (p<0,05) para los defectos espermáticos totales entre la primera (5,48) y la tercera colecta (1,08) en el G2. Los resultados sugieren que el desarrollo del aparato reproductor y la espermatogénesis están presentes a los 14 meses de edad.

Características seminais, corpóreas e anatômicas do aparelho reprodutor de reprodutores da raça canchim aos 14 e 48 meses de idade / Anatomical, corporal and seminal characteristics of the reproductive system of canchim breed reproductors at ages from 14 to 48 months / Características seminales, corpóreas y anatómicas del aparato reproductor de reproductores de la raza canchim a los 14 y 48 meses de edad

Foram utilizados 20 touros da raça Canchim em inatividade sexual, divididos em grupo 1 (G1), constituído de 10 animais com 14 meses de idade e grupo 2 (G2), com 10 animais de 48 meses de idade. A proposta desse estudo foi investigar as características dos ejaculados e das mensurações anatômicas do trato reprodutivo da raça Canchim. Os parâmetros obtidos foram: peso médio de 445,5 e 706,02 kg; circunferência escrotal de 31,80 e 36,25 cm e índice de massa corpórea de 270,33 e 346,73 kg/m2 para o G1 e G2, respectivamente, com diferenças significativas (p < 0,05) entre eles. Realizaram-se quatro colheitas de sêmen por touro por meio de eletroejaculação, com intervalos de 14 dias, verificando-se diferença significativa (p < 0,05) para os defeitos espermáticos totais, entre a primeira (5,48%) e a terceira colheitas (1,08%), no G2. Os resultados sugerem que o desenvolvimento do aparelho reprodutivo e a espermatogênese estão presentes aos 14 meses de idade.(AU)

Twenty bulls of the Canchin breed in sexual inactivity were divided in group 1(G1) with ten animals at the age of 14 months old and group 2 (G2) with ten animals at the age of 48 months old. The purpose of this study was to investigate the characteristics of the ejaculators and anatomic measurements of reproductive tract in Canchim breed. The parameters obtained were the average weight from 445.5 to 706.02 kg, scrotal circumference from 31.8 to 36.25 cm and Corporal Mass Index from 270.33 to 346.73 kg/m² for G1 and G2 respectively with a significant difference (p<0.05). Four semen collections per bull were made with electro ejaculation with intervals of 14 days, and it showed significant difference (p<0.05) for the total spermatic flaws, between the first (5.48%) and the third collect (1.08%) in G2. These results suggest that the development of the reproductive tract and the spermatogenesis are present at the age of 14 months old.(AU)

Fueron utilizados 20 toros de la raza Canchim en inactividad sexual, divididos en grupos:Grupo 1 (G1) con 10 animales, con edades de 14 meses y el grupo 2 (G2) con 10 animales, con edades de 48 meses, teniendo como objetivo verificar la adaptación al clima tropical, por medio de las características de los eyaculados y medidas anatómicas del aparato reproductivo. Los parámetros obtenidos fueron peso medio de 445,5 y 706,02 kg; circunferencia escrotal de 31,80 y 36,25 cm e índice de masa corpórea de 270,33 y 346,73 kg/m² para el G1 y G2, respectivamente, con diferencias significativas (p<0,05). Fueron realizadas cuatro colectas de semen por toro, con intervalo de 14 días, por medio de electroeyaculación verificando diferencias significativas (p<0,05) para los defectos espermáticos totales entre la primera (5,48%) y la tercera colecta (1,08%) en el G2. Los resultados sugieren que el desarrollo del aparato reproductor y la espermatogénesis están presentes a los 14 meses de edad.(AU)

Secreção de interferon-tau em embriões bovinos produzidos in vitro frescos e congelados / Interferon tau secretion in cattle embryos in vitro fertilized before and after cryopreservation

Avaliou-se a interferência da criopreservação sobre a secreção de interferon-tau (IFN-t) por embriões bovinos produzidos in vitro. Usaram-se dois grupos de tratamentos: I) constituído por embriões não criopreservados (fresco) e II) embriões criopreservados. Os embriões, após atingirem a fase de blastocisto (fresco ou imediatamente após o descongelamento dos criopreservados), continuaram a ser cultivados individualmente por mais sete dias. Do meio de cultivo em que se mantiveram os blastocistos retiraram-se alíquotas com três e sete dias do início do cultivo, para a avaliação da secreção de IFN-t pelos embriões cultivados. Os embriões congelados secretaram menos IFN-t do que aqueles não criopreservados(P<0,05), e com sete dias houve maior secreção do interferon do que com três dias (P<0,05). A criopreservação prejudicou a produção de IFN-t pelo trofoblasto e pode comprometer o reconhecimento materno da gestação e o desenvolvimento do embrião pós-descongelamento.

Secreção de interferon-tau em embriões bovinos produzidos in vitro frescos e congelados / Interferon tau secretion in cattle embryos in vitro fertilized before and after cryopreservation

Avaliou-se a interferência da criopreservação sobre a secreção de interferon-tau (IFN-t) por embriões bovinos produzidos in vitro. Usaram-se dois grupos de tratamentos: I) constituído por embriões não criopreservados (fresco) e II) embriões criopreservados. Os embriões, após atingirem a fase de blastocisto (fresco ou imediatamente após o descongelamento dos criopreservados), continuaram a ser cultivados individualmente por mais sete dias. Do meio de cultivo em que se mantiveram os blastocistos retiraram-se alíquotas com três e sete dias do início do cultivo, para a avaliação da secreção de IFN-t pelos embriões cultivados. Os embriões congelados secretaram menos IFN-t do que aqueles não criopreservados (P<0,05), e com sete dias houve maior secreção do interferon do que com três dias (P<0,05). A criopreservação prejudicou a produção de IFN-t pelo trofoblasto e pode comprometer o reconhecimento materno da gestação e o desenvolvimento do embrião pós-descongelamento.

The effect of cryopreservation in IFN-tau, from bovine embryos produced in vitro was evaluated. Two treated groups (G1= fresh bovine embryos, n=59 and G2= freezed embryos, n=84) were used to study the effect of cryopreservation on IFN-tau secretion. After reaching the blastocyst phase, the embryos were kept on individual culture for additional period of 7 days. On days 3 and 7 after the beginning of embryos cultivation, samples of the media culture were taken for IFN-tau secretion titration. Oocysts taken from follicles ranging from 3 to 5mm in diameter were obtained from ovaries of females at slaughterhouse. The embryos were frozen, after being dehydrated with ethylene glycol (1.8m), conditioned on 0.5ml palletes and frozen. Frozen embryos secreted lower IFN-tau than fresh embryos (P<0.05). At day 7 it was registered higher IFN-tau secretion from trophoblast than at day 3 (P<0.05). The increasing of IFN-tau secretion was observed when the blastocyst began to longed and it was directly related to the embryos development. The synthesis of IFN-tau is related to the capability of development of the blastocyst. Cryopreservation is a method that affects the maternal recognition of pregnancy and the post-freezing embryo development.

Secreção de interferon-tau em embriões bovinos produzidos in vitro frescos e congelados / Interferon tau secretion in cattle embryos in vitro fertilized before and after cryopreservation

Avaliou-se a interferência da criopreservação sobre a secreção de interferon-tau (IFN-t) por embriões bovinos produzidos in vitro. Usaram-se dois grupos de tratamentos: I) constituído por embriões não criopreservados (fresco) e II) embriões criopreservados. Os embriões, após atingirem a fase de blastocisto (fresco ou imediatamente após o descongelamento dos criopreservados), continuaram a ser cultivados individualmente por mais sete dias. Do meio de cultivo em que se mantiveram os blastocistos retiraram-se alíquotas com três e sete dias do início do cultivo, para a avaliação da secreção de IFN-t pelos embriões cultivados. Os embriões congelados secretaram menos IFN-t do que aqueles não criopreservados (P<0,05), e com sete dias houve maior secreção do interferon do que com três dias (P<0,05). A criopreservação prejudicou a produção de IFN-t pelo trofoblasto e pode comprometer o reconhecimento materno da gestação e o desenvolvimento do embrião pós-descongelamento.(AU)

The effect of cryopreservation in IFN-tau, from bovine embryos produced in vitro was evaluated. Two treated groups (G1= fresh bovine embryos, n=59 and G2= freezed embryos, n=84) were used to study the effect of cryopreservation on IFN-tau secretion. After reaching the blastocyst phase, the embryos were kept on individual culture for additional period of 7 days. On days 3 and 7 after the beginning of embryos cultivation, samples of the media culture were taken for IFN-tau secretion titration. Oocysts taken from follicles ranging from 3 to 5mm in diameter were obtained from ovaries of females at slaughterhouse. The embryos were frozen, after being dehydrated with ethylene glycol (1.8m), conditioned on 0.5ml palletes and frozen. Frozen embryos secreted lower IFN-tau than fresh embryos (P<0.05). At day 7 it was registered higher IFN-tau secretion from trophoblast than at day 3 (P<0.05). The increasing of IFN-tau secretion was observed when the blastocyst began to longed and it was directly related to the embryos development. The synthesis of IFN-tau is related to the capability of development of the blastocyst. Cryopreservation is a method that affects the maternal recognition of pregnancy and the post-freezing embryo development.(AU)

A continuous fluorescent assay for the determination of plasma and tissue angiotensin I-converting enzyme activity.

A continuous assay using internally quenched fluorescent peptides with the general sequence Abz-peptidyl-(Dnp)P-OH (Abz = ortho-aminobenzoic acid; Dnp = 2,4-dinitrophenyl) was optimized for the measurement of angiotensin I-converting enzyme (ACE) in human plasma and rat tissues. Abz-FRK(Dnp)P-OH, which was cleaved at the Arg-Lys bond by ACE, was used for the enzyme evaluation in human plasma. Enzymatic activity was monitored by continuous recording of the fluorescence (lambda ex = 320 nm and lambda em = 420 nm) at 37 degrees C, in 0.1 M Tris-HCl buffer, pH 7.0, with 50 mM NaCl and 10 microM ZnCl2. The assays can be performed directly in the cuvette of the fluorimeter and the hydrolysis followed for 5 to 10 min. ACE measurements in the plasma of 80 healthy patients with Hip-His-Leu and with Abz-FRK(Dnp)P-OH correlated closely (r = 0.90, P < 0.001). The specificity of the assay was demonstrated by the complete inhibition of hydrolysis by 0.5 microM lisinopril or captopril. Abz-FRK(Dnp)P-OH cleavage by ACE was monitored in rat lung, kidney, heart, and liver homogenates in the presence of a cocktail of inhibitors containing trans-epoxy-succinyl-L-leucylamido-(4-guanido)-butene, pepstatin, phenyl-methylsulfonyl fluoride, N-tosyl-L-phenylalanyl-chloromethyl ketone, and N-tosyl-lysyl-chloromethyl ketone to prevent undesirable hydrolysis. ACE activity in lung, heart and kidney homogenates, but not in liver homogenates, was completely abolished by 0.5 microM lisinopril or captopril. The advantages of the method are the procedural simplicity and the high sensitivity providing a rapid assay for ACE determinations.

A continuous fluorescent assay for the determination of plasma and tissue angiotensin I-converting enzyme activity

A continuous assay using internally quenched fluorescent peptides with the general sequence Abz-peptidyl-(Dnp)P-OH (Abz = ortho-aminobenzoic acid; Dnp = 2,4-dinitrophenyl) was optimized for the measurement of angiotensin I-converting enzyme (ACE) in human plasma and rat tissues. Abz-FRK(Dnp)P-OH, which was cleaved at the Arg-Lys bond by ACE, was used for the enzyme evaluation in human plasma. Enzymatic activity was monitored by continuous recording of the fluorescence (lambdaex = 320 nm and lambdaem = 420 nm) at 37°C, in 0.1 M Tris-HCl buffer, pH 7.0, with 50 mM NaCl and 10 æM ZnCl2. The assays can be performed directly in the cuvette of the fluorimeter and the hydrolysis followed for 5 to 10 min. ACE measurements in the plasma of 80 healthy patients with Hip-His-Leu and with Abz-FRK(Dnp)P-OH correlated closely (r = 0.90, P < 0.001). The specificity of the assay was demonstrated by the complete inhibition of hydrolysis by 0.5 æM lisinopril or captopril. Abz-FRK(Dnp)P-OH cleavage by ACE was monitored in rat lung, kidney, heart, and liver homogenates in the presence of a cocktail of inhibitors containing trans-epoxy-succinyl-L-leucylamido-(4-guanido)-butene, pepstatin, phenyl-methylsulfonyl fluoride, N-tosyl-L-phenylalanyl-chloromethyl ketone, and N-tosyl-lysyl-chloromethyl ketone to prevent undesirable hydrolysis. ACE activity in lung, heart and kidney homogenates, but not in liver homogenates, was completely abolished by 0.5 æM lisinopril or captopril. The advantages of the method are the procedural simplicity and the high sensitivity providing a rapid assay for ACE determinations.

IgA secretora total e específica no colostro e leite de mães da cidade do Natal-Rio Grande do Norte, Brasil / Total and specific IgA in colostrum and milk of mothers of Natal-Rio Grande do Norte, Brasil

PURPOSE: To determine the concentration of total secretory IgA and evaluate the repertoire of IgA antibodies to enteropathogenic Escherichia coli and Shigella flexneri antigens in colostrums and milk from mothers in Natal, RN. METHODS: The sample was constituted by 22 healthy clinically women whose babies were born at public hospital in Natal, RN. To determine total secretory IgA a radial immunedifusion tecnique (Mancini et al, 1965), was employed and to detect specific antibodies, immuneenzimatic assays, ELISA was used. RESULTS: The median values of total secretory IgA concentration presented individual variations with high levels in colostrums samples, decreasing during lactation, it was observed a p < 0.001 among the samples from the first day of lactation, to the thirtieth for total IgA concentration. All the donators present in colostrum and milk specific antibodies to Escherichia coli enteropathogenic (EPEC) and Shigella flexneri with titles higer in colostrum. There was parallel and directional pattern between total IgA and IgA anti-EPEC and Shegella flexneri, during period. CONCLUSION: The concentrations of total SIgA and specific antibodies to enteropathogenic Escherichia coli and Shigella flexneri in colostrums and milk in our study do not differ from others accomplished among populations with the same social and econimic features, stressing the importance of human milk as a protector agent against pathogens.

Desenvolvimento pós-fecundaçäo de oócitos bovinos pré-maturados em fluido folicular / Post-fertilization development of bovine oocytes pre-matured in follicular fluid

Estudou-se o efeito da pré-maturaçäo em fluido folicular bovino (FFb) sobre o potencial de desenvolvimento de ovócitos bovinos imaturos. Complexo cumulus- ovócitos (CCO) e FFb foram obtidos de ovários coletados em matadouro. O FFb foi inativado e os CCOs imaturos distribuídos em quatro tratamentos: (T1) 70 por cento de FFb em Talp Hepes, (T2) 100 por cento de FFb, (T3) 100 por cento de Talp Hepes, e (T4) controle. Em T1, T2 e T3 os CCOs foram pré - maturados por cinco horas a 37ºC em ar e posteriormente maturados in vitro. Em T4 a maturaçäo ocorreu logo após a aspiraçäo. Depois de fecundados in vitro, os ovócitos foram co-cultivados com células do cumulus por 10 dias. Avaliaram-se as taxas de clivagem, de produçäo de blastocistos no sétimo e oitavo dias pós- fecundaçäo (PF), de produçäo total e de blastocistos eclodidos no oitavo e nono dias PF. Calcularam-se as taxas de blastocistos no sétimo e oitavo dia e de blastocistos eclodidos em funçäo do total de blastocistos produzidos. As taxas de clivagem, de produçäo total e de blastocistos eclodidos näo diferiram entre os tratamentos (P>0,05), entretanto a produçäo de blastocistos no sétimo dia foi menor nos tratamentos com FFb e Talp Hepes (P<0,05). Concluiu-se que FFb e Talp Hepes na pré-maturaçäo por cinco horas atrasam o desenvolvimento embrionário sem comprometer a taxa de produçäo total de embriöes ou sua viabilidade.

Desenvolvimento pós-fecundação de oócitos bovinos pré-maturados em fluido folicular / Post-fertilization development of bovine oocytes pre-matured in follicular fluid

Estudou-se o efeito da pré-maturação em fluido folicular bovino (FFb) sobre o potencial de desenvolvimento de ovócitos bovinos imaturos. Complexo cumulus- ovócitos (CCO) e FFb foram obtidos de ovários coletados em matadouro. O FFb foi inativado e os CCOs imaturos distribuídos em quatro tratamentos: (T1) 70 por cento de FFb em Talp Hepes, (T2) 100 por cento de FFb, (T3) 100 por cento de Talp Hepes, e (T4) controle. Em T1, T2 e T3 os CCOs foram pré - maturados por cinco horas a 37ºC em ar e posteriormente maturados in vitro. Em T4 a maturação ocorreu logo após a aspiração. Depois de fecundados in vitro, os ovócitos foram co-cultivados com células do cumulus por 10 dias. Avaliaram-se as taxas de clivagem, de produção de blastocistos no sétimo e oitavo dias pós- fecundação (PF), de produção total e de blastocistos eclodidos no oitavo e nono dias PF. Calcularam-se as taxas de blastocistos no sétimo e oitavo dia e de blastocistos eclodidos em função do total de blastocistos produzidos. As taxas de clivagem, de produção total e de blastocistos eclodidos não diferiram entre os tratamentos (P>0,05), entretanto a produção de blastocistos no sétimo dia foi menor nos tratamentos com FFb e Talp Hepes (P<0,05). Concluiu-se que FFb e Talp Hepes na pré-maturação por cinco horas atrasam o desenvolvimento embrionário sem comprometer a taxa de produção total de embriões ou sua viabilidade.(AU)

The effect of the pre-maturation with bovine follicular fluid (bFF) on developmental competence of immature bovine oocytes was studied. Cumulus-oocytes complexes (COC) and bFF were obtained from ovaries collected at slaughterhouse. bFF was inactivated prior to use and COCs were distributed in four treatments: (T1) 70% of bFF in Talp hepes, (T2) 100% of bFF, (T3) 100% of Talp Hepes medium, and (T4) control group. In T1, T2, and T3, COCs were incubated during 5h at 37ºC before proceeding with in vitro maturation. In T4, maturation was performed soon after follicular aspiration. After in vitro fertilization, the presumptive zygotes were co-cultured with cumulus cells during 10 days. It was evaluated cleavage, blastocyst on seventh and eighth days post-fertilization (PF), overall blastocyst and hatched blastocyst on eighth and ninth days PF. It was also calculated the ratio of blastocyst on seventh and eighth days, and hatched blastocyst in function of overall blastocyst production. Cleavage, overall blastocyst and hatched blastocyst rates were similar (P>0.05) among treatments. However, blastocyst production on seventh day was lower (P<0.05) for bFF and Talp Hepes. These results indicate that bFF and Talp Hepes in pre-maturation for five hours caused a delay on embryonic development, although the overall blastocyst production and viability were not affected.(AU)

Taurina no desenvolvimento de embriöes bovinos fecundados in vitro / Taurine on the development of in vitro fertilized bovine embryos

O objetivo deste trabalho foi avaliar o efeito de diferentes concentraçöes de taurina no desenvolvimento de embriöes bovinos fecundados in vitro em meio de cultivo com diferentes fontes de soro. No experimento 1, zigotos (n=440) fecundados in vitro foram distribuídos aleatoriamente nos tratamentos com 0, 3, 7 ou 14 mM de taurina em meio de cultivo acrescido de 10 por cento de soro fetal bovino (SFB) e 3g/l de albumina sérica bovina (BSA). No experimento 2, os zigotos (n=940) foram divididos nos tratamentos com 0, 3 ou 14 mM de taurina em meio acrescido de 10 por cento de SFB ou 3g/l de BSA. No experimento 3, os zigotos (n=191) foram divididos nos tratamentos com 0 ou 3 mM de taurina em meio de cultivo sem fonte de soro, porém adicionado de 3 g/l de álcool polivinil. Nos experimentos 1 e 2 näo se observou diferença (P>0,05) na taxa de clivagem, na produçäo de blastocistos e no número de células entre as concentraçöes de taurina avaliadas. No experimento 3 encontraram-se maior (P<0,05) taxa de clivagem (68,5 por cento vs. 16,9 por cento) e produçäo de blastocistos (8,3 por cento vs. 0 por cento) na presença de taurina. O cultivo de zigotos em meio adicionado de SFB produziu maior (P<0,01) taxa de blastocistos no sétimo (25,6 por cento vs. 6,7 por cento) e oitavo (30,8 por cento vs. 13,9 por cento) dia pós-fecundaçäo e número de células/blastocistos (104,8± 2,63 vs. 84,7± 3,86) do que no cultivo com BSA, apesar de menor (P<0,01) taxa de clivagem (58,1 por cento vs. 71,3 por cento). Conclui-se que o efeito benéfico da taurina no desenvolvimento embrionário somente é observado na ausência de SFB e BSA. O SFB produz menor taxa de clivagem mas melhora o desenvolvimento embrionário após as primeiras divisöes celulares

Taurina no desenvolvimento de embriões bovinos fecundados in vitro

The effect of different taurine concentrations on bovine embryo development in medium supplemented with different serum sources was studied. In the first experiment, in vitro fertilized zygotes (n=440) were divided into treatments with 0, 3, 7 or 14 mM of taurine in culture medium supplemented with 10% of fetal calf serum (FCS) and 3g/l of bovine serum albumin (BSA). In the second experiment, zygotes (n=940) were divided into treatments with 0, 3 or 14 mM of taurine in cultured medium supplemented with 10% of FCS or 3g/l of BSA. In the third experiment, zygotes (n=191) were divided into treatments with 0 or 3 mM of taurine in culture medium without serum source, even so supplemented with 3g/l of polyvinyl alcohol. In the first and second experiments no differences (P>0.05) in cleavage rate, blastocyst production and cells number among the concentrations of taurine were observed. In the third experiment, taurine increased (P 0.05) cleavage rate (68.5% vs. 16.9%) and blastocysts production (8.3% vs. 0%). The culture of zygotes in medium supplemented with FCS produced more (P 0.01) blastocyst in the seventh (25.6% vs. 6.7%) and eighth (30.8% vs. 13.9%) day post-fertilization and total cells number/blastocysts (104.8± 2.63 vs. 84.7± 3.86) than in medium with BSA, despite lower (P 0.01) cleavage rate (58.1% vs. 71.3%). In conclusion, taurine only has a beneficial effect in the embryo development in culture medium in the absence of FCS and BSA. Fetal calf serum decreases cleavage rate, however, it improves the embryo development after the early cleavage.

O objetivo deste trabalho foi avaliar o efeito de diferentes concentrações de taurina no desenvolvimento de embriões bovinos fecundados in vitro em meio de cultivo com diferentes fontes de soro. No experimento 1, zigotos (n=440) fecundados in vitro foram distribuídos aleatoriamente nos tratamentos com 0, 3, 7 ou 14 mM de taurina em meio de cultivo acrescido de 10% de soro fetal bovino (SFB) e 3g/l de albumina sérica bovina (BSA). No experimento 2, os zigotos (n=940) foram divididos nos tratamentos com 0, 3 ou 14 mM de taurina em meio acrescido de 10% de SFB ou 3g/l de BSA. No experimento 3, os zigotos (n=191) foram divididos nos tratamentos com 0 ou 3 mM de taurina em meio de cultivo sem fonte de soro, porém adicionado de 3 g/l de álcool polivinil. Nos experimentos 1 e 2 não se observou diferença (P>0,05) na taxa de clivagem, na produção de blastocistos e no número de células entre as concentrações de taurina avaliadas. No experimento 3 encontraram-se maior (P 0,05) taxa de clivagem (68,5% vs. 16,9%) e produção de blastocistos (8,3% vs. 0%) na presença de taurina. O cultivo de zigotos em meio adicionado de SFB produziu maior (P 0,01) taxa de blastocistos no sétimo (25,6% vs. 6,7%) e oitavo (30,8% vs. 13,9%) dia pós-fecundação e número de células/blastocistos (104,8± 2,63 vs. 84,7± 3,86) do que no cultivo com BSA, apesar de menor (P 0,01) taxa de clivagem (58,1% vs. 71,3%). Conclui-se que o efeito benéfico da taurina no desenvolvimento embrionário somente é observado na ausência de SFB e BSA. O SFB produz menor taxa de clivagem mas melhora o desenvolvimento embrionário após as primeiras divisões celulares.

Taurina no desenvolvimento de embriões bovinos fecundados in vitro

The effect of different taurine concentrations on bovine embryo development in medium supplemented with different serum sources was studied. In the first experiment, in vitro fertilized zygotes (n=440) were divided into treatments with 0, 3, 7 or 14 mM of taurine in culture medium supplemented with 10% of fetal calf serum (FCS) and 3g/l of bovine serum albumin (BSA). In the second experiment, zygotes (n=940) were divided into treatments with 0, 3 or 14 mM of taurine in cultured medium supplemented with 10% of FCS or 3g/l of BSA. In the third experiment, zygotes (n=191) were divided into treatments with 0 or 3 mM of taurine in culture medium without serum source, even so supplemented with 3g/l of polyvinyl alcohol. In the first and second experiments no differences (P>0.05) in cleavage rate, blastocyst production and cells number among the concentrations of taurine were observed. In the third experiment, taurine increased (P 0.05) cleavage rate (68.5% vs. 16.9%) and blastocysts production (8.3% vs. 0%). The culture of zygotes in medium supplemented with FCS produced more (P 0.01) blastocyst in the seventh (25.6% vs. 6.7%) and eighth (30.8% vs. 13.9%) day post-fertilization and total cells number/blastocysts (104.8± 2.63 vs. 84.7± 3.86) than in medium with BSA, despite lower (P 0.01) cleavage rate (58.1% vs. 71.3%). In conclusion, taurine only has a beneficial effect in the embryo development in culture medium in the absence of FCS and BSA. Fetal calf serum decreases cleavage rate, however, it improves the embryo development after the early cleavage.

O objetivo deste trabalho foi avaliar o efeito de diferentes concentrações de taurina no desenvolvimento de embriões bovinos fecundados in vitro em meio de cultivo com diferentes fontes de soro. No experimento 1, zigotos (n=440) fecundados in vitro foram distribuídos aleatoriamente nos tratamentos com 0, 3, 7 ou 14 mM de taurina em meio de cultivo acrescido de 10% de soro fetal bovino (SFB) e 3g/l de albumina sérica bovina (BSA). No experimento 2, os zigotos (n=940) foram divididos nos tratamentos com 0, 3 ou 14 mM de taurina em meio acrescido de 10% de SFB ou 3g/l de BSA. No experimento 3, os zigotos (n=191) foram divididos nos tratamentos com 0 ou 3 mM de taurina em meio de cultivo sem fonte de soro, porém adicionado de 3 g/l de álcool polivinil. Nos experimentos 1 e 2 não se observou diferença (P>0,05) na taxa de clivagem, na produção de blastocistos e no número de células entre as concentrações de taurina avaliadas. No experimento 3 encontraram-se maior (P 0,05) taxa de clivagem (68,5% vs. 16,9%) e produção de blastocistos (8,3% vs. 0%) na presença de taurina. O cultivo de zigotos em meio adicionado de SFB produziu maior (P 0,01) taxa de blastocistos no sétimo (25,6% vs. 6,7%) e oitavo (30,8% vs. 13,9%) dia pós-fecundação e número de células/blastocistos (104,8± 2,63 vs. 84,7± 3,86) do que no cultivo com BSA, apesar de menor (P 0,01) taxa de clivagem (58,1% vs. 71,3%). Conclui-se que o efeito benéfico da taurina no desenvolvimento embrionário somente é observado na ausência de SFB e BSA. O SFB produz menor taxa de clivagem mas melhora o desenvolvimento embrionário após as primeiras divisões celulares.

Protective effect of thiourea, a hydroxyl-radical scavenger, on curcumin-induced chromosomal aberrations in an in vitro mammalian cell system.

Natural dietary antioxidants are extensively studied for their ability to protect cells from damage to DNA, protein, and lipids induced by antitumor agents or radiation that leads to the generation of free radical in normal cells in vivo and in vitro. Curcumin is a natural antioxidant known to possess therapeutic properties and has been reported to scavenge free radicals and to inhibit clastogenesis in mammalian cells. However, curcumin has been reported to induce a significant increase in the frequency of chromosomal aberrations in Chinese hamster ovary (CHO) cells. To investigate whether the clastogenic activity of curcumin in CHO cells in culture can be ascribed to a pro-oxidant behavior, mediated by free radical generation, experiments were carried out with the combination of curcumin (15 microg/ml) and thiourea (10, 20, or 40 microg/ml), a potent hydroxyl radical scavenger. The results showed that the clastogenic action of curcumin was statistically decreased in a dose-dependent manner in the presence of thiourea. These data have shown that curcumin-induced chromosomal damage in CHO cells can be mediated by hydroxyl radical generation in the present experimental conditions. Teratogenesis Carcinog. Mutagen. 21:175-180, 2001.

Vancomycin monitoring in term newborns: comparison of peak and trough serum concentrations determined by high performance liquid chromatography and fluorescence polarization immunoassay.

INTRODUCTION: Peak and trough serum concentrations of vancomycin were determined in term newborn infants with confirmed or suspected Staphylococcus sp sepsis by high performance liquid chromatography and flourescence polarization immunoassay. OBJECTIVE: To statistically compare the results of the high performance liquid chromatography and flourescence polarization immunoassay techniques for measuring serum vancomycin concentrations. METHODS: Eighteen peak and 20 trough serum samples were assayed for vancomycin concentrations using high performance liquid chromatography and flourescence polarization immunoassay from October 1995 to October 1997. RESULTS: The linear correlation coefficients for high performance liquid chromatography and flourescence polarization immunoassay were 0.27 (peak, P = 0.110) and 0.26 (trough, P = 0.1045) respectively, which were not statistically significant. CONCLUSION: There was wide variation in serum vancomycin concentrations determined by high performance liquid chromatography as compared with those determined by flourescence polarization immunoassay. There was no recognizable pattern in the variability; in an apparently random fashion, the high performance liquid chromatography measurement was sometimes substantially higher than the flourescence polarization immunoassay measurement, and at other times it was substantially lower.

Brain abscess by citrobacter diversus in infancy: case report.

Citrobacter diversus is closely related to brain abscess in newborn infants. We describe a case of brain abscess by this bacteria in a newborn infant and his clinical and cranial computed tomographic evaluation until the fourth month of life and discuss therapeutic management of this patient.