Selection of autochthonous lactic acid bacteria from goat dairies and their addition to evaluate the inhibition of Salmonella typhi in artisanal cheese.

This study aimed to select autochthonous lactic acid bacteria (LAB) with probiotic and functional properties from goat dairies and test their addition to artisanal cheese for the inhibition of Salmonella typhi. In vitro tests, including survival in the gastrointestinal tract (GIT), auto- and co-aggregation, the hemolytic test, DNase activity, antimicrobial susceptibility, antibacterial activity, tolerance to NaCl and exopolysaccharide (EPS), gas and diacetyl production were conducted for sixty isolates. Based on these tests, four LAB isolates (UNIVASF CAP 16, 45, 84 and 279) were selected and identified. Additional tests, such as production of lactic and citric acids by UNIVASF CAP isolates were performed in addition to assays of bile salt hydrolase (BSH), ß-galactosidase and decarboxylase activity. The four selected LAB produced high lactic acid (>17 g/L) and low citric acid (0.2 g/L) concentrations. All selected strains showed BSH and ß-galactosidase activity and none showed decarboxylase activity. Three goat cheeses (1, 2 and control) were produced and evaluated for the inhibitory action of selected LAB against Salmonella typhi. The cheese inoculated with LAB (cheese 2) decreased 0.38 log10 CFU/g of S. Typhy population while in the cheese without LAB inoculation (cheese 1) the pathogen population increased by 0.29 log units. Further, the pH value increased linearly over time, by 0.004 units per day in cheese 1. In the cheese 2, the pH value decreased linearly over time, by 0.066 units per day. The cocktail containing selected Lactobacillus strains with potential probiotic and technological properties showed antibacterial activity against S. typhi in vitro and in artisanal goat cheese. Thus, goat milk is important source of potential probiotic LAB which may be used to inhibit the growth of Salmonella population in cheese goat, contributing to safety and functional value of the product.

Concurrent Loss of Heterozygosity and Mosaic Deletion of 12p13.32pter.

Deletions in the short arm of chromosome 12 are the rarest subtelomeric imbalances. Less than 20 patients have been reported to date, and their microdeletions were identified either by FISH or array-CGH without SNP data. Here, we report a patient with a 12p13.32pter mosaic deletion detected by chromosome microarray analysis with loss of heterozygosity (LOH) of the deleted segment in addition to the adjacent distal segment. LOH is indicative of a complex rearrangement, suggestive of mitotic microhomology-mediated break-induced replication.

Partial 1q Duplications and Associated Phenotype.

Duplications of the long arm of chromosome 1 are rare. Distal duplications are the most common and have been reported as either pure trisomy or unbalanced translocations. The paucity of cases with pure distal 1q duplications has made it difficult to delineate a partial distal trisomy 1q syndrome. Here, we report 2 patients with overlapping 1q duplications detected by G-banding. Array CGH and FISH were performed to characterize the duplicated segments, exclude the involvement of other chromosomes and determine the orientation of the duplication. Patient 1 presents with a mild phenotype and carries a 22.5-Mb 1q41q43 duplication. Patient 2 presents with a pure 1q42.13qter inverted duplication of 21.5 Mb, one of the smallest distal 1q duplications ever described and one of the few cases characterized by array CGH, thus contributing to a better characterization of distal 1q duplication syndrome.

Microbiological and physical-chemical profile of goat milk in the semiarid region of the San Francisco valley / Perfil microbiológico e físico-químico do leite caprino na região semiárida do vale do São Francisco

In Brazil, the largest goat herd is in the Northeast, especially in Bahia and Pernambuco, which are the first and second states in number of goats. The semiarid region of San Francisco Valley still lacks studies on determination of parameters for the goat milk produced. The aim of this study was to characterize the microbiological and physical-chemical profile of goat milk produced in the semiarid region of the San Francisco Valley in accordance with current legislation. Forty-two samples of goat milk were obtained from six municipalities in the semiarid region of the San Francisco Valley (Juazeiro, Uauá, Senhor do Bonfim, Curaçá, Jaguarari and Petrolina), with seven samples per municipality, and the microbiological and physical-chemical parameters were analyzed. The microbiological analyses were Total Aerobic Mesophilic Bacteria (TAMB) and Lactic Acid Bacteria (LAB) counts, enumeration of total and thermotolerant coliforms and Escherichia coli research. Indirect measurement of Somatic Cell Count (SCC) was also performed. The physical-chemical tests performed were acidity, Freezing Point Depression (FDP) and quantitative determination of fat by the Gerber method. There were significant differences for microbiological analyses in the cities studied. In the samples, LAB was predominant among the microorganisms examined and E. coli was detected in all municipalities. For physicalchemical parameters, there was no significant difference only for FDP. The microbiological and physical-chemical characteristics were markedly separate by Principal Component Analysis (PCA). The cities of Petrolina and Juazeiro were characterised by greater quantification of fat and indirect SCC, respectively. In Uauá, Curaçá, Jaguarari and Senhor do Bonfim, the parameters for total coliforms, FDP, E. coli and acidity stood out, respectively. The microbiological results are indicative of the need for hygienic practices during milking to obtain better milk quality.

No Brasil, o maior rebanho de caprinos encontra-se no nordeste, com destaque para Bahia e Pernambuco, no qual são a primeira e segunda Unidade de Federação com o maior efetivo de caprinos. A região semiárida do Vale do São Francisco ainda é carente de estudos para fixar parâmetros para o leite caprino produzido. O objetivo deste trabalho foi caracterizar o perfil microbiológico e físicoquímico do leite caprino produzido na região semiárida do Vale do São Francisco em acordo com a legislação No Brasil, o maior rebanho de caprinos encontra-se no nordeste, com destaque para Bahia e Pernambuco, no qual são a primeira e segunda Unidade de Federação com o maior efetivo de caprinos. A região semiárida do Vale do São Francisco ainda é carente de estudos para fixar parâmetros para o leite caprino produzido. O objetivo deste trabalho foi caracterizar o perfil microbiológico e físicoquímico do leite caprino produzido na região semiárida do Vale do São Francisco em acordo com a legislação Petrolina e Juazeiro foram caracterizadas pela maior quantificação da gordura e contagem indireta de células somáticas, respectivamente. Em Uauá, Curaçá, Jaguarari e Senhor do Bonfim destacaram-se os parâmetros: coliformes totais, FDP, E. coli e acidez, respectivamente. Os resultados microbiológicos são indicativos da necessidade de uma melhor prática higiênica durante a ordenha para a obtenção de um leite de melhor qualidade.

Microbiological and physical-chemical profile of goat milk in the semiarid region of the San Francisco valley / Perfil microbiológico e físico-químico do leite caprino na região semiárida do vale do São Francisco

In Brazil, the largest goat herd is in the Northeast, especially in Bahia and Pernambuco, which are the first and second states in number of goats. The semiarid region of San Francisco Valley still lacks studies on determination of parameters for the goat milk produced. The aim of this study was to characterize the microbiological and physical-chemical profile of goat milk produced in the semiarid region of the San Francisco Valley in accordance with current legislation. Forty-two samples of goat milk were obtained from six municipalities in the semiarid region of the San Francisco Valley (Juazeiro, Uauá, Senhor do Bonfim, Curaçá, Jaguarari and Petrolina), with seven samples per municipality, and the microbiological and physical-chemical parameters were analyzed. The microbiological analyses were Total Aerobic Mesophilic Bacteria (TAMB) and Lactic Acid Bacteria (LAB) counts, enumeration of total and thermotolerant coliforms and Escherichia coli research. Indirect measurement of Somatic Cell Count (SCC) was also performed. The physical-chemical tests performed were acidity, Freezing Point Depression (FDP) and quantitative determination of fat by the Gerber method. There were significant differences for microbiological analyses in the cities studied. In the samples, LAB was predominant among the microorganisms examined and E. coli was detected in all municipalities. For physicalchemical parameters, there was no significant difference only for FDP. The microbiological and physical-chemical characteristics were markedly separate by Principal Component Analysis (PCA). The cities of Petrolina and Juazeiro were characterised by greater quantification of fat and indirect SCC, respectively. In Uauá, Curaçá, Jaguarari and Senhor do Bonfim, the parameters for total coliforms, FDP, E. coli and acidity stood out, respectively. The microbiological results are indicative of the need for hygienic practices during milking to obtain better milk quality.(AU)

No Brasil, o maior rebanho de caprinos encontra-se no nordeste, com destaque para Bahia e Pernambuco, no qual são a primeira e segunda Unidade de Federação com o maior efetivo de caprinos. A região semiárida do Vale do São Francisco ainda é carente de estudos para fixar parâmetros para o leite caprino produzido. O objetivo deste trabalho foi caracterizar o perfil microbiológico e físicoquímico do leite caprino produzido na região semiárida do Vale do São Francisco em acordo com a legislação No Brasil, o maior rebanho de caprinos encontra-se no nordeste, com destaque para Bahia e Pernambuco, no qual são a primeira e segunda Unidade de Federação com o maior efetivo de caprinos. A região semiárida do Vale do São Francisco ainda é carente de estudos para fixar parâmetros para o leite caprino produzido. O objetivo deste trabalho foi caracterizar o perfil microbiológico e físicoquímico do leite caprino produzido na região semiárida do Vale do São Francisco em acordo com a legislação Petrolina e Juazeiro foram caracterizadas pela maior quantificação da gordura e contagem indireta de células somáticas, respectivamente. Em Uauá, Curaçá, Jaguarari e Senhor do Bonfim destacaram-se os parâmetros: coliformes totais, FDP, E. coli e acidez, respectivamente. Os resultados microbiológicos são indicativos da necessidade de uma melhor prática higiênica durante a ordenha para a obtenção de um leite de melhor qualidade.(AU)

Transient neonatal hypothyroidism in a boy with unbalanced translocation t(8;16) / Hipotireoidismo neonatal transitório em um menino com translocação não balanceada t(8;16)

Genetic defects resulting in deficiency of thyroid hormone synthesis can be found in about 10% of the patients with permanent congenital hypothyroidism, but the identification of genetic abnormalities in association with the transient form of the disease is extremely rare. We report the case of a boy with transient neonatal hypothyroidism that was undiagnosed in the neonatal screening, associated with extrathyroid malformations and mental retardation. The boy carries an unbalanced translocation t(8;16), and his maternal uncle had a similar phenotype. Chromosomal analysis defined the patient's karyotype as 46,XY,der(8)t(8;16)(q24.3;q22)mat,16qh+. Array-CGH with patient's DNA revealed a ~80 kb terminal deletion on chromosome 8q24.3qter, and a ~21 Mb duplication on chromosome 16q22qter. ZNF252 gene, mapped to the deleted region on patient's chromosome 8, is highly expressed in the thyroid, and may be a candidate gene for our patient's transient neonatal thyroid dysfunction. This is the first report on the association of a chromosomal translocation with the transient form of congenital hypothyroidism. This description creates new hypothesis for the physiopathology of transient congenital hypothyroidism, and may also contribute to the definition of the unbalanced translocation t(8;16)(q24.3;q22) phenotype, which has never been described before. Arq Bras Endocrinol Metab. 2012;56(8):564-9.

Defeitos genéticos resultando em deficiência hormonal tireoidiana podem ser encontrados em cerca de 10% dos pacientes com hipotireoidismo congênito permanente, porém a identificação de anormalidades genéticas associadas à forma transitória da doença é extremamente rara. Relatamos o caso de um menino com hipotireoidismo neonatal transitório não diagnosticado no teste de triagem neonatal, associado a malformações extratireoidianas e retardo mental. O paciente é portador de translocação não balanceada t(8;16), e seu tio materno tinha fenótipo similar. A análise cromossômica definiu o cariótipo do paciente como 46,XY,der(8)t(8;16)(q24.3;q22)mat,16qh+. A análise cromossômica array-CGH com o DNA do paciente revelou deleção terminal de ~80 kb no cromossomo 8q24.3qter, e duplicação de ~21 Mb no cromossomo 16q22qter. O gene ZNF252, mapeado na região da deleção no cromossomo 8 do paciente, é altamente expresso na tireoide e pode ser um gene candidato no hipotireoidismo neonatal transitório do paciente. Esse é o primeiro relato de associação de uma translocação cromossômica com a forma transitória do hipotireoidismo congênito. Essa descrição descortina novas hipóteses para a fisiopatologia do hipotireoidismo congênito transitório e também pode contribuir para a definição do fenótipo da translocação não balanceada t(8;16)(q24.3;q22), nunca descrito anteriormente. Arq Bras Endocrinol Metab. 2012;56(8):564-9.

Does celiac disease occur in Afro-derived Brazilian populations?

BACKGROUND: Celiac disease is an autoimmune disorder that occurs in genetically susceptible individuals in whom the ingestion of dietary gluten induces intestinal mucosa inflammation. Previous studies suggest that celiac disease may either be very rare or underdiagnosed in African and/or African-derived population. AIM: Determine the prevalence of celiac disease in Sub-Saharan African-derived Brazilian communities using serological screening. SUBJECTS AND METHODS: Inhabitants from 10 African-derived communities from Northeastern of Brazil were screened for celiac disease. All sera were tested for endomysial class IgA antibody using indirect immunofluorescence. RESULTS: No positive test for IgA-endomysial was observed in the 860 individuals tested. CONCLUSION: Our data suggests a low prevalence of celiac disease in African-derived Brazilian populations.

Transient neonatal hypothyroidism in a boy with unbalanced translocation t(8;16).

Genetic defects resulting in deficiency of thyroid hormone synthesis can be found in about 10% of the patients with permanent congenital hypothyroidism, but the identification of genetic abnormalities in association with the transient form of the disease is extremely rare. We report the case of a boy with transient neonatal hypothyroidism that was undiagnosed in the neonatal screening, associated with extrathyroid malformations and mental retardation. The boy carries an unbalanced translocation t(8;16), and his maternal uncle had a similar phenotype. Chromosomal analysis defined the patient's karyotype as 46,XY,der(8)t(8;16)(q24.3;q22)mat,16qh+. Array-CGH with patient's DNA revealed a ~80 kb terminal deletion on chromosome 8q24.3qter, and a ~21 Mb duplication on chromosome 16q22qter. ZNF252 gene, mapped to the deleted region on patient's chromosome 8, is highly expressed in the thyroid, and may be a candidate gene for our patient's transient neonatal thyroid dysfunction. This is the first report on the association of a chromosomal translocation with the transient form of congenital hypothyroidism. This description creates new hypothesis for the physiopathology of transient congenital hypothyroidism, and may also contribute to the definition of the unbalanced translocation t(8;16)(q24.3;q22) phenotype, which has never been described before.

Aspectos clínicos da leucemia mieloide crônica em pacientes do Hospital de Base do Distrito Federal / Clinical features of chronic myeloid leukemia in patients at the Hospital de Base do Distrito Federal

Objetivo. Descrever aspectos clínicos, hematológicos e de sobrevida de pacientes com leucemia mieloide crônica em hospital especializado de Brasília, Brasil. Método. Estudo descritivo e retrospectivo de 223 pacientes atendidos no período de maio de 2002 a abril de 2009. Resultados. A mediana de idade foi 45 anos. Aproximadamente, 92% dos pacientes foram classificados como tendo doença em fase crônica, 5% em fase acelerada e 3% em crise blástica. Observou-se esplenomegalia em64,7% de 150 pacientes, inclusive as de grandes dimensões (29,3%), além de hiperleucocitose, com mediana 120.200/mm3, e elevada proporção de pacientes (67,5%) em grupos de risco intermediário e de alto risco de acordo com escore prognóstico de Sokal. A sobrevida global foi 84,7% aos oito anos, com intervalo de confiança de 95%:75,5% a 90,7%, e a sobrevida livre de progressão foi 73,5% no mesmo período com intervalo de confiança de 95%: 61,9% a 82%, embora diversas modalidades de tratamento tenham sido usadas. A sobrevida global aos oito anos foi significativamente maior em doentes do grupo de baixo risco no escore de Sokal (p = 0,017). Conclusão. Os pacientes se apresentaram com características de doença avançada ao diagnóstico, quando analisados em conjunto e foram seguidos em desfechos de longo prazo com resultados inferiores aos observados na literatura médica.

Objective. The authors described clinical, hematological and survival features of patients with chronic myeloid leukemia in a specialized hospital of Brasília, Brazil. Method. This is a retrospective and descriptive study which included 223 patients treated from May 2002 to April 2009.Results. The median age was 45 years. Approximately, 92% of patients were classified as having chronic phase, 5% accelerated phase and 3% in blast crisis. Splenomegaly was observed in a significant proportion of them, 64.7 % out of 150, and it was large in 29.3%. Hyperleucocytosis, median 120.200/mm3, was observed and a high proportion of patients(67.5%) were classified as having intermediate and high risk group according to Sokal prognostic score. The at eight years overall survival was 84.7%, with 95% confidence interval of 75.5% to 90.7%, and progression free survival was 73.5%, with 95% confidence interval of 61.9% to 82%, in the same period, although several treatment protocols were employed. Overall survival was significantly better in patients with low risk Sokal score (p = 0,017). Conclusion. The patients presented features of advanced disease at diagnosis and long-term outcomes were worse than those from published data in the medical literature.

Genotoxic effects of white fluorescent light on human lymphocytes in vitro.

Sources of light beams such as white fluorescent light, are present in our daily life to meet the needs of life in the modern world. This study was conducted with the objective of determining the possible genotoxic, cytotoxic and aneugenic effects caused by this agent in different stages of the cell cycle (G0/early G1, S, and late G2), using different cytogenetic parameters (sister chromatid exchanges--SCE, chromosome aberrations--CA, and detection of aneugenic effects) in lymphocytes from temporary cultures of human peripheral blood. WFL showed a genotoxic effect in vitro, expressed by an increase in the frequency of SCE's, regardless of the cell cycle stage. However, no increase in the frequency of CAs was observed. In addition, disturbances in cell cycle kinetics and chromosomal segregation were also observed. Taken together, such data may contribute to a better understanding and a different management in the use of phototherapy for some pathological conditions.

Influência do resfriamento na incidência de aneuploidia de ovócitos bovinos maturados in vitro / Influence of cooling on the incidence of aneuploidy of in vitro matured bovine oocytes

O objetivo do estudo foi verificar o efeito do resfriamento de ovócitos bovinos na incidência de aneuploidia. Ovócitos bovinos foram obtidos de ovários de abatedouro e divididos em cinco grupos: grupo controle (ovócitos não resfriados); grupo 0/4 (ovócitos resfriados a 4 °C antes do inicio da maturação); grupo 0/29 (ovócitos resfriados a 29 °C antes do inicio da maturação); grupo 12/4 (ovócitos resfriados a 4 °C após 12 de maturação); e grupo 12/29 (ovócitos resfriados a 29 °C após 12 horas de maturação). Os ovócitos permaneceram resfriados por 45 minutos. Em todos os grupos os ovócitos completaram 24 horas de maturação. Em seguida, os ovócitos foram fixados em lâminas e corados com orceína acética. Não foi observada diferença significativa (P > 0,05) na incidência de metáfase II aneuploides entre o grupo controle (0/21; 0%) e os grupos resfriados a 4 °C (25/26; 3,8%) e 29°C (27/29; 6,8%) antes do começo da maturação e 12 horas após o inicio da maturação (21/24; 12,5% e 27/30; 10%, respectivamente). Conclui-se que o resfriamento a 4 e 29 °C antes do início da maturação ou 12 após não afeta a taxa de aneuploidia, de ovócitos bovinos, após completarem 24 horas de maturação in vitro.

The objective of the study was to verify the incidence of bovine aneuploid oocytes when cooled. Bovine cumulus-oocyte complexes were recovered from ovaries at a slaughterhouse and then divided into five groups: control group (uncooled oocytes), 0/4 h group (composed of oocytes cooled before the onset of maturation at 4°C), 0/29 (composed of oocytes cooled before the onset of maturation at 29 °C), 12/4 (cooled 12 h after the onset of maturation at 4°C), and 12/29 h group (cooled 12 h after the onset of maturation at 29 °C). The oocytes remained cooled for 45 min. In all groups, the oocytes completed 24 h of maturation. Subsequently, the cumulus cells were removed, and the denuded oocytes fixed on slides and stained with aceto-orcein. No differences (P > 0.05) in the incidence of aneuploid metaphase II oocytes were observed between the control group (0/21; 0%) and oocytes cooled at 4°C and 29 °C before (3.8% and 6.8%) and after 12 h the onset of maturation (12.5% and 10%). It is concluded that the cooling at 4 and 29 °C before the maturity or 12 after not affect the rate of aneuploidy of bovine oocytes, after completing 24 hours of in vitro maturation.

INFLUENCE OF COOLING ON THE INCIDENCE OF ANEUPLOIDY OF IN VITRO MATURED BOVINE OOCYTES / INFLUÊNCIA DO RESFRIAMENTO NA INCIDÊNCIA DE ANEUPLOIDIA DE OVÓCITOS BOVINOS MATURADOS IN VITRO

The objective of the study was to verify the incidence of bovine aneuploid oocytes when cooled. Bovine cumulus-oocyte complexes were recovered from ovaries at a slaughterhouse and then divided into five groups: control group (uncooled oocytes), 0/4 h group (composed of oocytes cooled before the onset of maturation at 4ºC), 0/29 (composed of oocytes cooled before the onset of maturation at 29 ºC ), 12/4 (cooled 12 h after the onset of maturation at 4ºC), and 12/29 h group (cooled 12 h after the onset of maturation at 29 ºC). The oocytes remained cooled for 45 min. In all groups, the oocytes completed 24 h of maturation. Subsequently, the cumulus cells were removed, and the denuded oocytes fixed on slides and stained with aceto-orcein. No differences (P > 0.05) in the incidence of aneuploid metaphase II oocytes were observed between the control group (0/21; 0%) and oocytes cooled at 4ºC and 29 ºC before (3.8% and 6.8%) and after 12 h the onset of maturation (12.5% and 10%). It is concluded that the cooling at 4 and 29 C before the maturity or 12 after not affect the rate of aneuploidy of bovine oocytes, after completing 24 hours of in vitro maturation.

O objetivo do estudo foi verificar o efeito do resfriamento de ovócitos bovinos na incidência de aneuploidia. Ovócitos bovinos foram obtidos de ovários de abatedouro e divididos em cinco grupos: grupo controle (ovócitos não resfriados); grupo 0/4 (ovócitos resfriados a 4 ºC antes do inicio da maturação); grupo 0/29 (ovócitos resfriados a 29 ºC antes do inicio da maturação); grupo 12/4 (ovócitos resfriados a 4 ºC após 12 de maturação); e grupo 12/29 (ovócitos resfriados a 29 ºC após 12 horas de maturação). Os ovócitos permaneceram resfriados por 45 minutos. Em todos os grupos os ovócitos completaram 24 horas de maturação. Em seguida, os ovócitos foram fixados em lâminas e corados com orceína acética. Não foi observado diferença significativa (P > 0,05) na incidência de metáfase II aneuplóides entre o grupo controle (0/21; 0%) e os grupos resfriados a 4 ºC (25/26; 3,8%) e 29ºC (27/29; 6,8%) antes do começo da maturação e 12 horas após o inicio da maturação (21/24; 12,5% e 27/30; 10%, respectivamente). Conclui-se que o resfriamento a 4 e 29 ºC antes do início da maturação ou 12 após não afeta a taxa de aneuploidia, de ovócitos bovinos, após completarem 24 horas de maturação in vitro.

Effect of colling in diploid oocytes after in vitro maturation / Efeito do resfriamento na ploidia de ovócitos bovinos maturados in vitro

The present study aimed to verify the incidence of bovine diploid oocytes when cooled at various maturation stages. Bovine cumulus-oocyte complexes were recovered from ovaries at a local slaughterhouse and divided into five groups: control group (uncooled oocytes), 0/4 h group (oocytes cooled at 4ºC before the onset of maturation), 0/29 (composed of oocytes cooled at 29ºC before the onset of maturation), 12/4 (cooled 12 h at 4ºC after the onset of maturation), and 12/29 h group (cooled 12 h at 29ºC after the onset of maturation). the oocytes remained cooled for 45 min. in all groups, the oocytes completed 24 h maturation. Subsequently, the cumulus cells were removed, and the denuded oocytes fixed on slides and stained with aceto-orcein. No differences (P > 0.05) in the incidence of diploid metaphase ii oocytes were observed between the control group (6.0%) and oocytes cooled at 4ºC and 29ºC before (8.9% and 8.0%) and after 12 h the onset of maturation (3.9% and 0.0%). These results suggest that the nuclear stage at which bovine oocytes are cooled does not affect the incidence of diploid oocytes after 24 h maturation. Key-Words: Bovine, cooling, diploid, in vitro maturation, oocytes.

O presente estudo objetivou verificar o efeito do resfriamento de ovócitos bovinos em diferentes estágios de maturação na ploidia. Ovócitos bovinos foram obtidos de ovários de abatedouro e divididos em cinco grupos: grupo-controle (ovócitos não resfriados); grupo 0/4 (ovócitos resfriados a 4ºC antes do inicio da maturação); grupo 0/29 (ovócitos resfriados a 29ºC antes do início da maturação); grupo 12/4 (ovócitos resfriados a 4ºC após doze horas de maturação); e grupo 12/29 (ovócitos resfriados a 29ºC após doze horas de maturação). Os ovócitos permaneceram resfriados por 45 minutos. Em todos os grupos os ovócitos completaram 24 horas de maturação. Em seguida, as células da corona radiata foram removidas e os ovócitos fixados em lâminas e corados com orceína acética. Não se observou diferença significativa (P>0,05) na incidência de metáfase II diplóide entre o grupo-controle (6,0%) e os grupos resfriados a 4ºC (8,9%) e 29ºC (8,0%) antes do começo da maturação e doze horas após o inicio da maturação (3,9% e 0,0%, respectivamente). Os resultados sugerem que resfriamento em diferentes períodos de maturação não afeta a ploidia de ovócitos bovinos após completarem 24 horas de maturação in vitro. Palavras-chaves: Bovinos, diplóide, ovócitos, maturação in vitro, resfriamento.

EFFECT OF COLLING IN DIPLOID OOCYTES AFTER IN VITRO MATURATION / EFEITO DO RESFRIAMENTO NA PLOIDIA DE OVÓCITOS BOVINOS MATURADOS IN VITRO

The present study aimed to verify the incidence of bovine diploid oocytes when cooled at various maturation stages. Bovine cumulus-oocyte complexes were recovered from ovaries at a local slaughterhouse and divided into five groups: control group (uncooled oocytes), 0/4 h group (oocytes cooled at 4ºC before the onset of maturation), 0/29 (composed of oocytes cooled at 29ºC before the onset of maturation), 12/4 (cooled 12 h at 4ºC after the onset of maturation), and 12/29 h group (cooled 12 h at 29ºC after the onset of maturation). the oocytes remained cooled for 45 min. in all groups, the oocytes completed 24 h maturation. Subsequently, the cumulus cells were removed, and the denuded oocytes fixed on slides and stained with aceto-orcein. No differences (P > 0.05) in the incidence of diploid metaphase ii oocytes were observed between the control group (6.0%) and oocytes cooled at 4ºC and 29ºC before (8.9% and 8.0%) and after 12 h the onset of maturation (3.9% and 0.0%). These results suggest that the nuclear stage at which bovine oocytes are cooled does not affect the incidence of diploid oocytes after 24 h maturation. Key-Words: Bovine, cooling, diploid, in vitro maturation, oocytes.

O presente estudo objetivou verificar o efeito do resfriamento de ovócitos bovinos em diferentes estágios de maturação na ploidia. Ovócitos bovinos foram obtidos de ovários de abatedouro e divididos em cinco grupos: grupo-controle (ovócitos não resfriados); grupo 0/4 (ovócitos resfriados a 4ºC antes do inicio da maturação); grupo 0/29 (ovócitos resfriados a 29ºC antes do início da maturação); grupo 12/4 (ovócitos resfriados a 4ºC após doze horas de maturação); e grupo 12/29 (ovócitos resfriados a 29ºC após doze horas de maturação). Os ovócitos permaneceram resfriados por 45 minutos. Em todos os grupos os ovócitos completaram 24 horas de maturação. Em seguida, as células da corona radiata foram removidas e os ovócitos fixados em lâminas e corados com orceína acética. Não se observou diferença significativa (P>0,05) na incidência de metáfase II diplóide entre o grupo-controle (6,0%) e os grupos resfriados a 4ºC (8,9%) e 29ºC (8,0%) antes do começo da maturação e doze horas após o inicio da maturação (3,9% e 0,0%, respectivamente). Os resultados sugerem que resfriamento em diferentes períodos de maturação não afeta a ploidia de ovócitos bovinos após completarem 24 horas de maturação in vitro. Palavras-chaves: Bovinos, diplóide, ovócitos, maturação in vitro, resfriamento.

GATA1 mutations in acute leukemia in children with Down syndrome.

It has been reported that somatic mutations in the X-linked GATA1 gene are present in hematological clonal disorders in children with Down syndrome (DS). We analyzed retrospective samples of DS children with acute myeloid leukemia, transient leukemia (TL), and myelodysplastic syndrome (MDS) to test whether the specificity of GATA1 mutations can be helpful in distinguishing these hematopoietic disorders. A total of 49 samples were subjected to GATA1 mutation screening by direct sequencing and denaturing polyacrylamide gel electrophoresis (PAGE). Mutations in exon 2 of GATA1 were detected in six of eight DS-AML M7 samples and in four of six DS-TL; no mutation was detected in 13 children with acute lymphoblastic leukemia (DS-ALL), 6 with DS-AML (M0, M2, and M5), 6 with DS-MDS and in 8 DS infants without hematological disorders and 2 children with AML M7 without DS. Blast cells proportion in the sample represented a critical aspect on the sensitivity of mutation detection in GATA1, and a combination of sequence analysis and PAGE is necessary to detect mutations when blast percentage is low. The absence of detected mutations in any of the DS-MDS cases raises the question whether MDS in DS children is an intermediate stage between TL and AML M7, as previously suggested.