Function of the pentose phosphate pathway and its key enzyme, transketolase, in the regulation of the meiotic cell cycle in oocytes / 대한생식의학회지

OBJECTIVE: Previously, we identified that transketolase (Tkt), an important enzyme in the pentose phosphate pathway, is highly expressed at 2 hours of spontaneous maturation in oocytes. Therefore, this study was performed to determine the function of Tkt in meiotic cell cycle regulation, especially at the point of germinal vesicle breakdown (GVBD). METHODS: We evaluated the loss-of-function of Tkt by microinjecting Tkt double-stranded RNAs (dsRNAs) into germinal vesicle-stage oocytes, and the oocytes were cultured in vitro to evaluate phenotypic changes during oocyte maturation. In addition to maturation rates, meiotic spindle and chromosome rearrangements, and changes in expression of other enzymes in the pentose phosphate pathway were determined after Tkt RNA interference (RNAi). RESULTS: Despite the complete and specific knockdown of Tkt expression, GVBD occurred and meiosis was arrested at the metaphase I (MI) stage. The arrested oocytes exhibited spindle loss, chromosomal aggregation, and declined maturation promoting factor and mitogen-activated protein kinase activities. The modified expression of two enzymes in the pentose phosphate pathway, Prps1 and Rbks, after Tkt RNAi and decreased maturation rates were amended when ribose-5-phosphate was supplemented in the culture medium, suggesting that the Tkt and pentose phosphate pathway are important for the maturation process. CONCLUSION: We concluded that Tkt and its associated pentose phosphate pathway play an important role in the MI-MII transition of the oocytes' meiotic cell cycle, but not in the process of GVBD.

Maturação biológica e desempenho físico de jovens atletas de ginástica rítmica / Biological maturation and physical performance of young athletes in rhythmic gymnastics

Objetivo: analisar a maturação biológica e desempenho da força em jovens atletas de ginástica rítmica. Materiais e Métodos: A amostra foi composta por nove moças da seleção paraibana (idade cronológica 12,44±3,00 anos), tempo de prática de 45,66+30,45meses submetidas aos testes físicos de: força de membros inferiores (dinamometria dorsal, salto horizontal e vertical) e membros superiores (dinamometria manual e medicineball) e testes antropométricos de: massa corporal, imc, altura total, envergadura, estatura, estadiamento de maturação sexual (EMS), por meio de auto-avaliação, pelas fotografias de Tanner, maturação óssea pelo método de Greulich-Pyle em Raio X com laudo médico no diagnóstico da idade. Resultados: Encontrou-se: massa corporal 45,9+5,6 kg; estatura 1,56±0,02 m; altura total 2,06±0,04 cm; envergadura 1,63±0,05 cm; IMC 18,6+1,8 kg/m2; idade óssea 15,9±2,2 anos e cronológica de 13,28±12,44 anos, apresentando diferenças significativas (p=0,001); EMS Púbis 2,33±1,22e Mama 3,11±0,93, sem diferenças significativas (p=0,065), sendo 33,3% pré-púberes e 66,7% púberes; o desempenho físico de salto horizontal foi de 150,52±10,01 cm, vertical 217,77±25,55 cm; força de membros superiores: arremesso de medicine ball 221±0,52cm; força em dinamometria manual 22,44±6,89 kg/f e dorsal 54,33±18,44 kg/f. Não foi encontrado diferenças entre os graus de maturação sexual e óssea e desempenho nos testes (p>0,076). Conclusão: A idade óssea é maior do que a cronológica, sendo que as maturações sexuais e ósseas não causaram impacto no desempenho da força.

Objective: To analyze the bone and sexual maturation, and strength performance in young athletes in rhythmic gymnastics. Methods: The sample consisted of nine girls of the Paraiba team (chronological age 12.44 +3.00 years), with practice time of 45.66 +30.45 months, subjected to physical tests: lower limb strength (dorsal dynamometry, horizontal and vertical jump) and upper limbs (handgrip and medicineball) and anthropometric tests: of body mass, BMI, height, wingspan, overall height, stage of sexual maturation (SSM), through self-assessment by photographs of Tanner, bone maturation by the method of Greulich-Pyle in X-ray medical report with the diagnosis of age. Results: There was a body mass 45.9±5.6 kg; height 1.56±0.02 m; overall height of 2.06±0.04 cm; wingspan 1.63±0.05 cm; BMI 18.6±1,8kg/m2, bone age 15.9 ± 2.2 years and chronological 13.28 ± 12.44 years, with significant differences (p=0.001); SSM Pubis 2.33±1.22 and Mama 3,11±0.93, without significant differences (p=0.065), being that 33.3% were prepubertal and 66.7% are pubescent; the physical performance of horizontal jump was 150.52±10.01 cm, Vertical 217.77±25.55 cm; strength of upper limbs: medicine ball throw of 221±0.52 cm; handgrip strength in 22.44±6.89 kg/f and dorsal 54.33±18.44 kg/f. No differenceswere found between the degree of sexual maturation and bone test performance (p> 0.076). Conclusion:The bone age is greater than the chronological, and the sexual maturation and bone caused no impact on the performance of the force.

Análise da relação entre a maturação biológica e estresse na coordenação motora grossa em escolares de 5 a 10 anos / Analysis of the relationship between stress and biological maturation in motor coordination in large school of 5 to 10 years

A infância é o período em que o desenvolvimento motor está sendo construído, onde, o aparecimento e a extensão do desenvolvimento de habilidades na fase de movimentos dependem de muitos fatores dos estímulos externos do ambiente. A coordenação motora é a capacidade do cérebro de equilibrar os movimentos do corpo, mais especificamente dos músculos e das articulações. Pode-se verificar o desempenho motor de uma pessoa através de sua agilidade, velocidade e energia. O objetivo da pesquisa foi identificar a interação entre maturação biológica, coordenação motora grossa e o estresse em escolares. A amostra contou com 380 crianças de 5 a 10 anos de idades de escolas públicas do município de Cacoal/RO. Para verificar a coordenação motora utilizou-se o teste de TGMD-2, para o teste de estresse infantil foi utilizado o Inventário de Estresse Infantil e para a maturação utilizou-se o protocolo de Tanner adaptado. Para a análise da normalidade amostral, foi utilizado o teste de Kolmogorov-Smirnov, sendo utilizado o programa SPSS 17,0. Através das medidas de tendência central e média, foi realizado a caracterização das variáveis medidas e das escalas medidas contínuas. Para verificar a interação das variáveis dependentes e independentes foi utilizado o Modelo Geral Linear (GLM). Sabe-se que o desenvolvimento motor sofre influências diretas de vários fatores, sendo esses genéticos, ambientais e psicológicos. Influencias essas que são um processo contínuo, onde acabam por provocar variações individuais, tornando-se assim único a cada indivíduo.

Childhood is the time when the motor development is being built where the appearance and extent of skills development in the movement phase is dependent on many factors of external stimuli from the environment. Motor coordination is the brain's ability to balance the body's movements, specifically the muscles and joints. You can check the motor performance of a person through their agility, speed andpower. The objective of the research was to identify the interaction between biological maturation, gross motor coordination and stress in schoolchildren. The sample consisted of 380 children 5-10 years of ageattending public schools Cacoal / RO. TGMD-2 test was used to verify motor coordination and to evaluate children stress an inventory was used, maturation was tested by the adapted protocol of Tanner. For the analysis of normal sample, we used the Kolmogorov-Smirnov, and used SPSS 17.0. Through the measuresof central tendency and mean, was conducted to characterize the variables measured and continuous measures of scales. To verify the interaction of independent and dependent variables, we used the GeneralLinear Model (GLM). It is known that motor development suffers direct influences of various factors, and these genetic, environmental and psychological. These influences are a continuous process, whereeventually cause individual variations, thus making it unique to each individual.

The effects of protooncogene on oocyte maturation mediated by cytokines / 中国应用生理学杂志

<p><b>AIM</b>The mechanisms of cytokines in regulating oocyte maturation is still little known. The present study attempt to investigate whether the protooncogene of c-erbB2, c-myb are involved in introducing of cytokines to regulate oocyte maturation.</p><p><b>METHODS</b>This research used mouse GV stage oocyte culture model in vitro and RT-PCR, Western blotting method to explore the effect of EGF, TNFalpha, ET-1 and NO on oocyte maturation; to analyze the c-erbB2 mRNA and c-myb mRNA expression and the phosphorylation of MAPK and cyclinB1 expression in oocytes affected by above cytokines.</p><p><b>RESULTS</b>EGF(10 microg/L) stimulated meiosis of oocytes significantly, the level of c-erbB2 mRNA, c-myb mRNA were increased, and promoted the phosphorylation of MAPK and cyclinB1 expression; TNFalpha (1 microg/L) and ET-1 ((10(-1) mol/L) had the results to EGF. Low dose of SNP (10(-5)mol/L) had no effect on oocyte maturation, but could significantly reverse the suppression of dbcAMP on oocyte maturation.</p><p><b>CONCLUSION</b>c-erbB2 and c-myb were involved in introducing of cytokines to regulate oocyte maturation, might be the middle link in connection of the cytokines with MAPK and MPF in regulation oocyte maturation.</p>

Cdc25B overexpression reverses two-cell block in mouse embryos / 中国应用生理学杂志

<p><b>AIM</b>To explore the effect of Cdc25B overexpression on the development of mouse two-cell embryos.</p><p><b>METHODS</b>The pBSK-Cdc25B was in vitro transcribed into 5'-capped mRNA for microinjection by using mMESSAGE mMACHINE kit. The Cdc25B mRNA was microinjected into mouse embryos at two-cell stage in order to observe the embryonic development and cleavage rate. Using protein kinase activity assay and Western blot to detect the MPF activity as well as the phosphorylation status of Cdc2-Tyr15 in Cdc25B overexpression group respectively.</p><p><b>RESULTS</b>The mouse embryos with Cdc25B overexpression developed to the four-cell stage 48 h after the hCG injection with the percentage of cleavage over 40% compared with the embryos in control groups which still remained at the two-cell stage. Moreover, MPF activity increased significantly after Cdc25B mRNA injection. The phosphorylation status of Cdc2-Tyr15 was coincident with MPF activity.</p><p><b>CONCLUSION</b>The results indicate that Cdc25B overexpression in early mouse two-cell embryos reverses two-cell block and promotes their development into four-cell stage by activating MPF.</p>

c-erbB(2) and c-myb induce oocyte maturation via activation of mitogen-activated protein kinase and maturation promoting factor / 生理学报

It is important to study the mechanism of oocyte maturation because oocyte maturation is essential for the female procreation. The present study was designed to observe the effects of protooncogenes c-erbB(2) and c-myb on oocyte maturation and the upstream and downstream relationship with mitogen-activated protein kinase (MAPK) and maturation promoting factor (MPF). The investigation was designed as follows: (1) In order to explore the effects of protooncogenes on oocyte maturation, the dose- and time-dependent effects of c-erbB(2) antisense oligodeoxynucleotide (ASODN) and c-myb ASODN on oocyte maturation were examined, and the effects of oocyte microinjection with recombinant c-erbB(2) and c-myb proteins on oocyte maturation were investigated; (2) In order to study the upstream and downstream relationship among protooncogenes of c-erbB(2), c-myb and protein kinases of MAPK and MPF in regulating oocyte maturation, mouse oocytes were cultured in the medium treated with c-erbB(2) ASODN, c-myb ASODN, PD98059 (the MAPK inhibitor) or roscovitine (the MPF inhibitor) for 8 h, respectively, and the expressions of c-erbB(2) mRNA, c-myb mRNA, MAPK and MPF were examined. The results showed that both c-erbB(2) ASODN and c-myb ASODN inhibited the rate of germinal vesicle breakdown (GVBD) and the first polar (PB1) extrusion of denuded oocytes (DOs) in a dose- and time-dependent way, and delayed their maturation time significantly. When recombinant c-erbB(2) and c-myb proteins were microinjected into cytoplasm of germinal vesicle stage oocyte, we found that the GVBD rate increased by 23.1% (P<0.05) and 32.2% (P<0.05), respectively, for 6-hour culture, and the PB1 extrusion rate increased by 17.3% (P<0.05) and 23.5% (P<0.05), respectively, for 12-hour culture. RT-PCR showed that the mRNA expressions of c-erbB(2) and c-myb were detected in oocytes; c c-erbB(2) ASODN inhibited c-erbB(2) mRNA and c-myb mRNA expressions; c-myb ASODN inhibited c-myb mRNA expression but had no effect on c-erbB(2) mRNA expression. Nonsense tat ODN had no effects on the expressions of c-erbB(2) mRNA and c-myb mRNA. Neither PD98059 nor roscovitine changed the expressions of c-erbB(2) mRNA and c-myb mRNA though both of them inhibited recombinant c-erbB(2) and c-myb proteins-induced oocyte maturation. Furthermore, MAPK phosphorylation and cyclin B1 synthesis in oocytes were inhibited remarkably when oocytes were treated with c-erbB(2) ASODN, c-myb ASODN, PD98059 and roscovitine. Nonsense tat ODN had no effects on MAPK phosphorylation and cyclinB1 content. The results suggest that protooncogenes c-erbB(2) and c-myb play an important role in oocyte maturation; the effects of c-erbB(2) and c-myb depend upon the action of MAPK and MPF, and their activation is the event that occurs downstream of c-erbB(2) and c-myb in the maturation signal pathway.

In silico Analysis of Downstream Target Genes of Transcription Factors / 대한불임학회지

OBJECTIVE: In the previous study, we compiled the differentially expressed genes during early folliculogenesis.1 Objective of the present study was to identify downstream target genes of transcription factors (TFs) using bioinformatics for selecting the target TFs among the gene lists for further functional analysis. MATERIALS AND METHODS: By using bioinformatics tools, constituent domains were identified from database searches using Gene Ontology, MGI, and Entrez Gene. Downstream target proteins/genes of each TF were identified from database searches using TF database (TRANSFAC(R) 6.0) and eukaryotic promoter database (EPD). RESULTS: DNA binding and trans-activation domains of all TFs listed previously were identified, and the list of downstream target proteins/genes was obtained from searche of TF database and promoter database. Based on the known function of identified downstream genes and the domains, 3 (HNF4, PPARg, and TBX2) out of 26 TFs were selected for further functional analysis. The genes of wee1-like protein kinase and p21WAF1 (cdk inhibitor) were identified as potential downstream target genes of HNF4 and TBX2, respectively. PPARg, through protein-protein interaction with other protein partners, acts as a transcription regulator of genes of EGFR, p21WAF1, cycD1, p53, and VEGF. Among the selected 3 TFs, further study is in progress for HNF4 and TBX2, since wee1-like protein kinase and cdk inhibitor may involved in regulating maturation promoting factor (MPF) activity during early folliculogenesis. CONCLUSIONS: Approach used in the present study, in silico analysis of downstream target genes, was useful for analyzing list of TFs obtained from high-throughput cDNA microarray study. To verify its binding and functions of the selected TFs in early folliculogenesis, EMSA and further relevant characterizations are under investigation.

Interação entre células do cumulus e atividade da proteína quinase C em diferentes fases da maturação nuclear de oócitos bovinos / Interaction between cumulus cells and the activity of protein kinase C at different stages of bovine oocyte nuclear maturation

Verificou-se a influência da proteína quinase C (PK-C) no reinício e na progressão da meiose em oócitos bovinos, determinando se as células do cumulus são mediadoras da PK-C na regulação da maturação dos oócitos. Complexos cumulus-oócitos (CCO) e oócitos desnudos (OD), distribuídos aleatoriamente em seis tratamentos (T) com base na presença de um ativador da PK-C (PMA) (T1 e T2), de um forbol éster incapaz de ativar a PK-C (4alfa-PDD-controle) (T3 e T4) ou de apenas o meio básico (TCM-199-controle) (T5 e T6), foram cultivados por 7, 9, 12, 18 e 22 horas. A percentagem de rompimento da vesícula germinativa no grupo cultivado com PMA foi maior do que nos dois grupos controle, com e sem células do cumulus. O cultivo de CCO e OD por 12 e 18 horas demonstrou que a PK-C influencia a progressão para os estádios de metáfase I (MI) e metáfase II (MII) de maneira dependente das células do cumulus. Nos períodos de 9 e 22 horas, não foi possível observar diferença entre os grupos quanto aos diferentes estádios de maturação. A ativação da PK-C acelera o reinício da meiose independentemente das células somáticas e acelera a progressão até os estádios de MI e MII na dependência das células do cumulus.

The activity of M-phase promoting factor in oral normal tissue and tumor / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the content and activity of M-phase promoting factor (MPF) in pleomorphic adenoma, mucoepidermoid carcinoma, buccal carcinoma and normal tissue, in order to evaluate the role of MPF in the development of tumor and the relationship between MPF and malignant degree.</p><p><b>METHODS</b>The content and activity of MPF were assessed by immunobloting and Gollicano method.</p><p><b>RESULTS</b>The cdc2 and cyclinB (two subunits of MPF) were found both in normal and tumor tissues, and their content in tumor was higher than normal tissues. Buccal carcinoma was 64% higher than normal tissues. The activity of MPF in carcinoma was higher than normal tissue and had positive relation with the malignant extent.</p><p><b>CONCLUSIONS</b>The content and activity of MPF in tumor are higher than normal tissue. PKC can activate MPF. These results show PKC may promote tumor proliferation by activating MPF and also, the activity of MPF has some relation with malignant extent.</p>

Atenuación térmica de cultivos lácticos destinados a la maduración acelerada de quesos / Attenuation thermophilus of lactic cultivation destinated maturation acelerated of cheese

En el presente trabajo se evalúa el efecto de la atenuación mediante choque térmico a varias temperaturas de cultivos de streptococcus salivarius ssp. thermophilus y lactobacillus delbrueckii ssp. bulgaricus destinados a la maduración acelerada de queso tipo Dambo. El objetivo fue establecer condiciones adecuadas para el tratamiento térmico, suficiente para reducuir la producción de ácido láctico, sin dañar el sistema enzimático proteolítico importante para la maduración del queso. Suspensiones de bacterias se cultivaron a pH constante y se calentaron a 63, 67, 70 y 72ºC. Los cambios en el pH y en la actividad enzimática mediante el método del ácido trinitrobencenosulfónico se evaluaron para cada temperatura de atenuación. Las células calentadas a 70 y 72ºC produjeron ácido lentamente después de un período de retraso de horas, mientras que las células calentadas a 65ºC lo hicieron rápidamente, con un retraso de 2 horas. Los cultivos calentados a 63ºC se comportaron de forma similar al control la actividad proteolítica disminuyó en 30, 70, 83 y 93 por ciento respectivamente, comparada con la de las células sin ningún tratamiento

Efecto de la clonidina en la velocidad de crecimiento en niños y adolescentes con retardo constitucional de maduración: Barquisimeto 1995-97 / Effect by the clonidine at the velocity of growth in boys with constitucional delay growth: Barquisimeto 1995-97

Se hizo seguimiento, cada 3 meses durante un año, de 34 niños entre la edad de 9 a 14 años, que ingresaron a la Clínica de Crecimiento y Desarrollo del Hospital "Antonio María Pineda" de Barquisimeto, lapso 1995-97, con el diagnóstico de talla baja familiar y retardo constitucional de maduración; diecisiete recibieron clonidina a la dosis de 0.1 mg por metro cuadrado de superficie corporal/día durante 6 meses y los diecisiete restantes formaron el grupo control. Se calculó la velocidad de crecimiento del grupo en estudio durante los seis meses previos al tratamiento y seis meses durante el mismo. Se analizó la velocidad de crecimiento del grupo control y la velocidad de crecimiento del grupo estudio, previo al tratamiento (basal) y durante el suministro de clonidina. Se aplicó T de Student y sus factores predictivos: regresión lineal múltiple, encontrándose un incremento significativo de la velocidad de crecimiento (P< 0.004) durante la administración de clonidina al compararla con la velocidad previa (basal); igualmente fue significativamente mayor al relacionarla con el grupo control (P<0.02)

Avaliaçäo da atividade citotóxica, antiviral e antimicrobiana de Apocynaceae amazônicas / Evaluation of the activity cytotoxic, antiviral and antimicrobial of Apocynaceae Amazonian

A necessidade de se introduzir novos fármacos quimioterápicos é patente. Para tanto, a modelagem molecular, a terapia gênica e os produtos naturais têm sido os caminhos escolhidos para a obtenção de novas moléculas. As florestas brasileiras estão entre os principais celeiros de biodiversidade, grande parte não estudada do ponto de vista fitoquímico e farmacológico. Isso implica em grandes possibilidades de identificação de novos fármacos, uma vez que a riqueza da biodiversidade biológica pode ser refletida na riqueza da biodiversidade química. Trinta e oito extratos provenientes de espécies de Apocynaceae foram submetidos a um estudo de triagem farmacológica antiviral, antimicrobiana e citotoxicidade, in vitro...

Participación de los factores de crecimiento durante el desarrollo y maduración folicular en el ovario del mamifero / Participation of growth factors during follicular development and maturation in the mammal ovary

El progreso del desarrollo y la maduración folicular requiere de la participación en conjunto de diferentes moduladores del crecimiento tales como: gonadotropinas, hormonas esteroides, interleucinas y factores de crecimiento, en este caso tratamos los aspectos relacionados a los factores de crecimiento, su efecto en la regulación de la mitosis y la diferenciación de los componentes celulares del folículo, mediante acciones autócrinas y/o parácrinas. La acción sinérgica de los factores de crecimiento (EGF, TGFa,TGFß, FGF e IGFs) en la estimulación de la mitosis, está dada por un mecanismo de mutuo reforzamiento de sus actividades además de su interacción con las gonadotropinas y con las hormonas esteroides, favoreciendo con ello la proliferación y citodiferenciación del folículo, al estimular la producción y activación de enzimas esteroidogénicas y la utilización de colesterol provenientes de las lipoproteínas de alta y baja densidad, regulando de esta manera la disponibilidad de colesterol, que es el sustrato común para las hormonas esteroides producidas por las células de la granulosa y de la teca durante la maduración folicular

Human gastrointestinal sphincters: study on the maturation of the nitric oxide innervation

Estudos histoqyímicos relevam uma inervaçäo noradrenérgica dos esfincteres gastrointestinais mais densa do que as regiöes näo-esfincterianas. também tem sido mostrado uma rica rede interconectante de fibras nervosas óxido-nitrérgicas inibitórias adivindas da gânglia mioentérica e se distribuindo dentro da camada muscular circular, especialmente nas regioSes esfincterianas. a presente investigaçäo estudou, no intestino humano em desenvolvimento, a histomorfometria da inervaçäo óxido-nitrérgica diárias intestinais selecionadas, particularmente, das regiSes esfincterianas. Segmnetos da junçäo gastro-esofagiana, regiäo gastro-piloro-duodenal, regiäo íleo-cecal e reto distal de 14 fetos de idade gestacional entre 12 e 23 semanas, foram usados para mapeamento histoquímico da nicotinamida adenosina de óxido nuleotídeo fosfato diaforase. Imagens de secçSes randonizadas foram selecionadas para histofometria, usando um sistema computadorizado de análise de imagens. A partir dos resultados, as seguintes conclusSes foram tiradas: 1- existe uma rede muito rica em nervos óxido-nitrérgicos interconectado a gânglia mioentérica e a camada muscular circular de todos os níveis selecionados. sendo mais densa na junçäo gastro-esofagiana, piloro, junçäo íleo-cecal e esfincter anal interno; 2- existe uma correlaçäo linear negativa entre a atividade neuronal mioentérica óxido-nitrérgica (densidade ganglionar) e a idade gestacional, a qual pode ser expressa pela equaçäo: Densidade ganglionar = 30,158- 1,0313 x idade gestacional. O esôfago, o piloro e esfíncter anal interno foram as regiöes com as mais baixas densidades ganglionares. Esses achados sugerem que a inervaçäo óxodo-nitrergica naqulas áreas, associada com densidade ganglionares relativamente baixas, torna possível levantar a hipótese de ue a normalidade ou retardo da maturaçäo desta inervaçäo inibitória poderia estar envolvida na patogênes de algumas anomalias congênitas como a estemose hipertrófica do piloro e acalásia do esfíncter anal interno