RESUMO
Glycosphingolipid biosynthesis-globo series pathway genes (FUT1, FUT2, ST3GAL1, HEXA, HEXB, B3GALNT1, and NAGA) play an important regulatory role in the defense against Escherichia coli F18 in piglets. In this study, we identified the transcription initiation site and promoter of this gene cluster by mined previous RNA-seq results using bioinformatics tools. The FUT1 transcription initiation region included five alternative splicing sites and two promoter regions, whereas each of the six other genes had one promoter. Dual luciferase reporter results revealed significantly higher transcriptional activity by FUT1 promoter 2, indicating that it played a more important role in transcription. The promoters of glycosphingolipid biosynthesis genes identified contained a CpG island within the first 500 bp, except for the B3GALNT1 promoter which included fewer CpG sites. These results provide a deeper insight into methylation and the regulatory mechanisms of glycosphingolipid biosynthesis-globo series pathway genes in piglets.
Assuntos
Fucosiltransferases/genética , Glicoesfingolipídeos/biossíntese , Regiões Promotoras Genéticas , Suínos/genética , Animais , Ilhas de CpG , Metilação de DNA , Fucosiltransferases/metabolismo , Ativação Transcricional , Galactosídeo 2-alfa-L-FucosiltransferaseRESUMO
The cucumber expansin 10 (CsEXP10) gene was previously cloned from young cucumber fruits but its role has not been defined. To determine the role of this gene in plant growth and development, a CsEXP10 gene transformation system was established. The open reading frame of the gene was inserted behind the CaMV35S promoter of vector pCAMBIA1301, and the construct was introduced into tomato plants by Agrobacterium-mediated transformation. In total, 19 kanamycin-positive lines were produced and nine independent transgenic lines were identified by ß-glucuronidase and polymerase chain reaction (PCR) analysis. Quantitative real-time PCR analysis showed that levels of the CsEXP10 transcript were higher in transgenic lines than in a non-transgenic line.
Assuntos
Agrobacterium tumefaciens/genética , Proteínas de Plantas/genética , Solanum lycopersicum/genética , Transformação Genética , Regulação da Expressão Gênica de Plantas , Ordem dos Genes , Vetores Genéticos/genética , Plantas Geneticamente ModificadasRESUMO
The super antibiotic bactericidal/permeability-increasing (BPI) protein is a member of a new generation of proteins that have been implicated as endotoxin-neutralizing agents. In this study, recombinant porcine BPI protein was obtained by generating porcine BPI encoding prokaryotic, eukaryotic, and yeast expression vectors. Recombinant protein expression was detected in yeast GS115, Escherichia coli, and 293-6E cells by gel electrophoresis and Western blotting. Escherichia coli F18 is the primary Gram-negative bacteria in the gut and the main pathogen leading to diarrhea and edema dis-ease in weaning piglets. Therefore, E. coli F18-resistant and -sensitive Sutai piglets were used to test differential expression of BPI protein by Western blotting and to investigate the potential correlation between BPI protein expression and E. coli F18-susceptibility. Recombinant porcine BPI protein expression was not detected in the prokaryotic and yeast expression systems; however, soluble protein was detected in the eukaryotic expression system. These data indicate the strong bacterio-static action of the BPI protein and confirm the feasibility of obtaining large amounts of recombinant porcine BPI recombinant protein using this eukaryotic expression system. In addition, the BPI protein expres-sion levels in the E. coli F18-resistant group were significantly higher than those in the sensitive group, indicating that high BPI protein ex-pression is associated with resistance to E. coli F18. Our findings pro-vide a basis for further investigations into the development of a drug designed to confer resistance to E. coli F18 in weaning piglets.
Assuntos
Peptídeos Catiônicos Antimicrobianos/biossíntese , Proteínas Sanguíneas/biossíntese , Resistência à Doença/genética , Infecções por Escherichia coli/genética , Escherichia coli/genética , Animais , Peptídeos Catiônicos Antimicrobianos/genética , Proteínas Sanguíneas/genética , Suscetibilidade a Doenças/microbiologia , Suscetibilidade a Doenças/veterinária , Endotoxinas/genética , Endotoxinas/metabolismo , Escherichia coli/patogenicidade , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/patologia , Infecções por Escherichia coli/veterinária , Vetores Genéticos , Genótipo , Suínos , DesmameAssuntos
Cesárea/métodos , Complemento C3a/imunologia , Embolia Amniótica , Lúpus Eritematoso Sistêmico , Complicações na Gravidez , Adulto , Ativação do Complemento , Embolia Amniótica/diagnóstico , Embolia Amniótica/etiologia , Embolia Amniótica/imunologia , Embolia Amniótica/terapia , Feminino , Ruptura Prematura de Membranas Fetais/cirurgia , Humanos , Lúpus Eritematoso Sistêmico/complicações , Lúpus Eritematoso Sistêmico/imunologia , Lúpus Eritematoso Sistêmico/fisiopatologia , Trabalho de Parto Prematuro/cirurgia , Gravidez , Complicações na Gravidez/imunologia , Complicações na Gravidez/fisiopatologia , Resultado do Tratamento , Contração UterinaRESUMO
The guinea pig is an excellent animal model for studying reproductive biology of adult humans and most domestic animals. Yet, whether this animal might serve as a good model for embryonic stage investigations and determinations of signals affecting or directing ovary development remains unknown. These questions were addressed by examining morphological evolution and the expression of biomarkers of cell proliferation and apoptosis in the ovaries of fetal and neonatal guinea pigs in the present study. Embryonic and neonatal guinea pigs at 30, 40, 50, 60, and 68 days postcoitum (dpc) and at 1 day postpartum (dpp) were evaluated, and the dynamic changes in follicles between 30 dpc and 1 dpp were observed. Results also showed that a critical period of follicular development in guinea pig embryos occurred at 40 to 50 dpc. Moreover, the proliferating-cell nuclear antigen, a cell proliferation marker, immunohistochemically stained healthy follicles, while caspase-3, an apoptosis marker, was mainly observed in atretic follicles. Together, these results demonstrate that cell proliferation and apoptosis contribute to follicular formation, development, and atresia in fetal and neonatal guinea pig ovaries. Furthermore, this study confirmed that the guinea pig is also an excellent animal model for studying reproductive biology in human and domestic animal embryos.
Assuntos
Apoptose/genética , Proliferação de Células , Desenvolvimento Embrionário/genética , Ovário/crescimento & desenvolvimento , Animais , Feminino , Feto/metabolismo , Cobaias , Humanos , Ovário/metabolismoRESUMO
Choriocarcinoma is a highly malignant disease of trophoblastic cells, which affects young women in the reproductive years. The main sites of metastasis from choriocarcinoma are lung, vagina, liver, gastrointestinal tract and kidneys, and the involvement of the ovaries is extremely rare. The diagnosis of ovarian metastasis is made mainly by two-dimensional ultrasonography (2DUS) with color Doppler, which shows a large vessel in the center of the mass. The three-dimensional power Doppler ultrasound (3D power Doppler) and dynamic contrast-enhanced magnetic resonance imaging (MRI) are new diagnostic modalities not yet published in literature. We report a case of metastatic choriocarcinoma with left ovary involvement in a 48-year-old woman with history of molar pregnancy and irregular follow-up of this disease. We emphasize the main findings by 2DUS with color Doppler, 3D power Doppler and dynamic contrast-enhanced MRI. 3D power Doppler is able to improve the assessment of anatomical relationships of vessels with the ovarian mass, while the resonance angiography allows us to evaluate the anatomic relations of the mass and adjacent structures, as well as the iliac vessels. The 3D power Doppler and dynamic contrast-enhanced MRI are promising methods in the evaluation of metastasis arising from gestational trophoblastic tumors.
RESUMO
No período de janeiro de 1980 a dezembro de 1989 foram atendidas na Escola Paulista de Medicina, um centro de referência, 75 pacientes com mola hidatiforme, das quais 65 se submeteram a esvaziamento em nosso serviço, e dez em outros. Para o cálculo da freqüência da mola hidatiforme utilizamos somente as 65 pacientes cujo esvaziamento molar se procedeu na Escola Paulista de Medicina em relaçäo a 13.986 gravidezes (partos, abortos e gravidezes sectópicas) ocorridas no período estudado. Deste modo, a freqüência da mola hidatiforme foi de 1:215 gravidezes, valor comparável aos países asiáticos e africanos
Assuntos
Humanos , Feminino , Gravidez , Mola Hidatiforme/epidemiologia , Brasil/epidemiologia , IncidênciaRESUMO
Between January 1980 and December 1989, at Escola Paulista de Medicina, a referral centre, 75 patients with hydatiform mole were assisted. The frequency of the hydatiform mole was calculated in 65 patients, whose treatment was performed in our Hospital, out of 13,986 pregnancies, which had occurred in this period of study. Thus, the incidence was 1:215 pregnancies, similar to that found in Asian and African countries.