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OBJECTIVE: We aimed to simplify our fetal RHD genotyping protocol by changing the method to attach Illumina's sequencing adaptors to PCR products from the ligation-based method to a PCR-based method, and to improve its reliability and robustness by introducing unique molecular indexes, which allow us to count the numbers of DNA fragments used as PCR templates and to minimize the effects of PCR and sequencing errors. RESULTS: Both of the newly established protocols reduced time and cost compared with our conventional protocol. Removal of PCR duplicates using UMIs reduced the frequencies of erroneously mapped sequences reads likely generated by PCR and sequencing errors. The modified protocols will help us facilitate implementing fetal RHD genotyping for East Asian populations into clinical practice.
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Diagnóstico Prenatal , Sistema del Grupo Sanguíneo Rh-Hr , Alelos , Femenino , Genotipo , Humanos , Embarazo , Atención Prenatal , Reproducibilidad de los Resultados , Sistema del Grupo Sanguíneo Rh-Hr/genéticaRESUMEN
Differentially methylated regions (DMRs) have been widely explored as epigenetic biomarkers. Here, we developed a novel approach combining methylation-sensitive restriction enzyme (MSRE) and next-generation sequencing (NGS) to identify DMRs between chorionic villi (CV) and maternal blood cells (MBC). During NGS library preparation, adapter-ligated genomic DNA of CV and MBC were digested with the MSRE, HpaII, and PCR-amplified. As unmethylated HpaII sites were cleaved, the resulted library should contain only methylated HpaII sites. By sequencing both HpaII-digested CV and MBC libraries, 9 differentially methylated-HpaII sites on chromosome 21 which exhibited more than 50% methylation increase in CV were identified. These DMRs are epigenetic biomarkers to tell the difference between CV and MBC. Our approach will also be applicable to screen various tissue-specific epigenetic biomarkers.
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Metilación de ADN , Enzimas de Restricción del ADN/metabolismo , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Adulto , Células Sanguíneas/metabolismo , Vellosidades Coriónicas/metabolismo , Cromosomas Humanos Par 21/genética , ADN/química , ADN/metabolismo , Femenino , Biblioteca de Genes , Humanos , Reacción en Cadena de la Polimerasa , EmbarazoRESUMEN
The value of the cardio-ankle vascular index (CAVI) increases with age. All large-scale studies of the CAVI have investigated patients <80 years old. Thus, the clinical characteristics of high CAVI in patients aged 80 or more remain unclear. Therefore, we investigated (1) the CAVI in very elderly patients and (2) the determinants of a high CAVI in high-risk patients, including very elderly patients. The Cardiovascular Prognostic Coupling Study in Japan (Coupling Registry) is a prospective observational study of Japanese outpatients with any cardiovascular risk factors. We enrolled 5109 patients from 30 institutions (average age 68.7 ± 11.4 years, 52.4% males). We investigated the determinants of the CAVI by separating the patients into three groups: 970 middle-aged (<60 years), 3252 elderly (60-79 years), and 887 very elderly (≥80 years) patients. The CAVI values of the males were significantly higher those of the females in all age groups (<60 years: 7.81 ± 1.11 vs. 7.38 ± 0.99, P < .001; 60-79 years: 9.20 ± 1.29 vs. 8.66 ± 1.07, P < .001; ≥80 years: 10.26 ± 1.39 vs. 9.51 ± 1.12, P < .001). In all age groups, the CAVI of the patients with diabetes/glucose tolerance disorder was higher than that of the patients without diabetes/glucose tolerance disorder (<60 years: 7.82 ± 1.22 vs 7.58 ± 1.03, P = .002; 60-79 years: 9.23 ± 1.20 vs 8.78 ± 1.19, P < .001; ≥80 years: 10.04 ± 1.24 vs 9.75 ± 1.32, P = .002). The determinants of the CAVI in these very elderly patients were age, male sex, low BMI, and mean blood pressure. Diabetes/glucose tolerance disorder and glucose were independently associated with the CAVI in the patients aged <60 years and 60-79 years, but not in those aged ≥80 years after adjusting for other covariates.
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Enfermedades Cardiovasculares , Hipertensión , Rigidez Vascular , Anciano , Anciano de 80 o más Años , Tobillo , Índice Tobillo Braquial , Presión Sanguínea , Enfermedades Cardiovasculares/diagnóstico , Enfermedades Cardiovasculares/epidemiología , Femenino , Factores de Riesgo de Enfermedad Cardiaca , Humanos , Japón/epidemiología , Masculino , Persona de Mediana Edad , Pronóstico , Sistema de Registros , Factores de RiesgoRESUMEN
Advances in prenatal molecular testing have made it possible to diagnose most genetic disorders early in gestation. In utero mesenchymal stem cell (MSC) therapy can be a powerful tool to cure the incurable. With this in mind, this method could ameliorate potential physical and functional damage. However, the presence of maternal T cells trafficking in the fetus during pregnancy is thought to be the major barrier to achieving the engraftment into the fetus. We investigated the possibility of using maternal adipose-derived stem cells (ADSCs) for in utero transplantation to improve engraftment, thus lowering the risk of graft rejection. Herein, fetal brain engraftment using congenic and maternal ADSC grafts was examined via in utero stem cell transplantation in a mouse model. ADSCs were purified using the mesenchymal stem cell markers, PDGFRα, and Sca-1 via fluorescence-activated cell sorting. The PDGFRα+Sca-1+ ADSCs were transplanted into the fetal intracerebroventricular (ICV) at E14.5. The transplanted grafts grew for at least 28 days after in utero transplantation with PDGFRα+Sca-1+ ADSC, and mature neuronal markers were also detected in the grafts. Furthermore, using the maternal sorted ADSCs suppressed the innate immune response, preventing the infiltration of CD8 T cells into the graft. Thus, in utero transplantation into the fetal ICV with the maternal PDGFRα+Sca-1+ ADSCs may be beneficial for the treatment of congenital neurological diseases because of the ability to reduce the responses after in utero stem cell transplantation and differentiate into neuronal lineages.
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OBJECTIVE: We examined whether maternal serum cytokine profiles of mothers with early-onset fetal growth restriction (FGR) were associated with delivery within 2 weeks after sampling during the third trimester. STUDY DESIGN: This exploratory prospective cross-sectional study included a total of 20 singleton fetuses with early-onset FGR and 31 healthy controls. Maternal serum samples during the early third trimester were analyzed for 23 cytokines. RESULTS: Of 20 fetuses with early-onset FGR, 14 had delivery within 2 weeks after sampling. Multivariate analysis revealed that maternal serum concentrations of soluble vascular endothelial growth factor receptor-1 (sVEGFR-1) and soluble CD40 ligand (sCD40L) were independently associated with delivery within 2 weeks in early-onset FGR. Among cases of early-onset FGR, concentrations of almost all maternal serum cytokines were similar. Maternal serum sVEGFR-1 concentrations were high when delivery occurred within 2 weeks. Maternal serum sCD40L concentrations were elicited only in cases in which delivery within 2 weeks occurred due to fetal deterioration. CONCLUSION: We identified two biomarkers, one specific for FGR and the other dependent on severity, that were significant components of angiogenic activities and inflammation factors. Imbalances in serum protein expression may have a substantial effect on the pathogenesis of FGR.
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Ligando de CD40/sangre , Cesárea , Citocinas/sangre , Retardo del Crecimiento Fetal/sangre , Trabajo de Parto Inducido , Preeclampsia/sangre , Receptor 1 de Factores de Crecimiento Endotelial Vascular/sangre , Adulto , Biomarcadores/sangre , Peso al Nacer , Estudios de Casos y Controles , Estudios Transversales , Procedimientos Quirúrgicos Electivos , Endoglina/sangre , Femenino , Factor de Crecimiento Similar a EGF de Unión a Heparina/sangre , Humanos , Recién Nacido , Recién Nacido Pequeño para la Edad Gestacional , Leptina/sangre , Masculino , Análisis Multivariante , Factor de Crecimiento Placentario/sangre , Embarazo , Tercer Trimestre del Embarazo , Nacimiento Prematuro , Factores de Tiempo , Factor A de Crecimiento Endotelial Vascular/sangreRESUMEN
Vascular biomarkers, including the cardio-ankle vascular index (CAVI), are increasingly being recognized as important indicators of cardiovascular risk. CAVI has been shown to have good discriminative ability for detecting new-onset hypertension, but results of studies investigating cardiovascular risk prediction are inconsistent. Furthermore, there is a lack of data on the prognostic value of changes in CAVI over time. The Cardiovascular Prognostic Coupling study was designed to determine the impact of baseline CAVI and changes in CAVI on cardiovascular events in a Japanese cohort. The design of the ongoing, multicenter, prospective, observational registry and baseline characteristics of the enrolled population are reported. Eligible consecutive patients were aged ≥30 years, had ≥1 cardiovascular risk factor, and were being treated according to relevant Japanese guidelines. The primary outcome is time to onset of a major cardiovascular event (a composite of cerebral infarction, cerebral hemorrhage, subarachnoid hemorrhage, stroke of unknown etiology, myocardial infarction, cardiovascular intervention for angina pectoris, and sudden death). Screening and enrollment occurred over a period of 3 years, followed by ≥7 years of follow-up, with CAVI determined annually. A total of 5279 patients were registered, of whom 5109 had baseline data available and will be included in future analyses. Mean CAVI at baseline was 8.8 ± 1.4. The proportion of patients with CAVI of <8, 8-10 or >10 was 25.3%, 57.0%, and 17.7%, respectively. Data from this registry should provide information on the significance of baseline CAVI and change in CAVI as indicators of cardiovascular prognosis in a representative patient population.
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Enfermedades Cardiovasculares , Hipertensión , Rigidez Vascular , Anciano , Enfermedades Cardiovasculares/diagnóstico , Enfermedades Cardiovasculares/epidemiología , Humanos , Japón/epidemiología , Pronóstico , Estudios Prospectivos , Sistema de Registros , Factores de RiesgoRESUMEN
Objectives: To evaluate the usefulness of color Doppler in fetal cardiac ultrasound screening in the second trimester.Methods: Fetuses who underwent ultrasound screening at 18-20 weeks' gestation at Showa University Hospital between 2011 and 2016 were evaluated. After delivery, neonatal congenital heart abnormalities were reviewed and compared with the antenatal ultrasound findings. Since 2014, we have added color Doppler to the routine B mode evaluation of the fetal heart. Congenital heart diseases (CHDs) found antenatally and postnatally were compared before and after protocol alternation. Medical records of all fetuses who underwent ultrasound screening at 18-20 weeks' gestation at Showa University Hospital between 2011 and 2016 were retrospectively reviewed.Results: There were 47 cases of CHDs confirmed postnatally. The detection rates of CHDs were 45.0% (9/20) in 2011-2013 and 55.6% (15/27) in 2014-2016. In 2011-2013, cases with antenatal diagnosis showed obvious abnormal findings of three-vessel view and four-chamber view with the B mode. In 2014-2016, the detection rate of isolated ventricular septal defect (VSD) was elevated from 10 to 42.9% using color Doppler.Conclusions: In this study, color Doppler improved the detection rate of CHDs. Color Doppler could give us additional information on blood flow although the canal or vessel is too small to detect morphological changes in the second trimester. It might be a useful tool for screening of CHDs with stenosis, regurgitation, and shunt that are difficult to detect by only the B mode in the second trimester.
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Cardiopatías Congénitas , Ultrasonografía Prenatal , Femenino , Corazón Fetal/diagnóstico por imagen , Cardiopatías Congénitas/diagnóstico por imagen , Humanos , Recién Nacido , Embarazo , Segundo Trimestre del Embarazo , Diagnóstico Prenatal , Estudios Prospectivos , Estudios RetrospectivosRESUMEN
BACKGROUND: To avoid hemolytic disease of the fetus and newborn resulting from maternal alloantibodies against fetal Rh antigens, anti-D immunoglobulin is routinely administered to RhD-negative pregnant women in Japan. Fetal RHD genotyping using cell-free DNA may prevent unnecessary antibody administration; however, current PCR-based methods, which detect RHD deletion, do not address the higher rates of RHD-positive D antigen-negative alleles in nonwhite populations without additional inspections. METHODS: We developed an amplicon-sequencing method that could estimate the type of paternally inherited fetal RHD allele from 4 major RHD alleles in the Japanese population: the D antigen-positive allele (RHD*01, 92.9%) and 3 D antigen-negative alleles (RHD*01N.01, 6.6%; RHD*01EL.01, 0.3%; RHD*01N.04, 0.1%) using cell-free DNA obtained from the blood plasma of pregnant women. RESULTS: The method correctly determined the fetal RhD type even when RhD-negative pregnant women possessed an RHD-positive D antigen-negative allele: RHD*01EL.01 or RHD*01N.04. CONCLUSIONS: This method is a reliable noninvasive fetal RHD genotyping method for Japanese and other East Asian populations. The genotyping principle of amplifying 2 different regions using the same primer pair and distinguishing them by their sequence difference during the subsequent mapping procedure is also theoretically applicable to RHD-positive D antigen-negative alleles prevalent in Africans. Therefore, this method offers an opportunity to consider targeted administration of anti-D immunoglobulin to RhD-negative pregnant women in East Asian and African countries and to increase the specificity of the fetal RHD genotyping implemented nationwide in several European countries.
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Técnicas de Genotipaje , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Diagnóstico Prenatal/métodos , Sistema del Grupo Sanguíneo Rh-Hr/genética , Alelos , Pueblo Asiatico/genética , Ácidos Nucleicos Libres de Células , Femenino , Frecuencia de los Genes , Humanos , Recién Nacido , Embarazo , Sistema del Grupo Sanguíneo Rh-Hr/sangreRESUMEN
OBJECTIVES: To investigate the effects of mold-fermented cheese (MFC) on brain-derived neurotrophic factor (BDNF) in community-dwelling older Japanese women with mild cognitive impairment (MCI). DESIGN: Randomized controlled crossover trial. INTERVENTION: Participants were randomly assigned to 2 groups. The MFC group was provided with 33.4 g MFC (camembert cheese) daily for 3 months, and the non-MFC group was provided with the same amount of non-MFC (processed cheese made from mozzarella cheese and cream cheese) for 3 months. After the post-intervention analysis (primary analysis), there was a 3-month washout period, followed by a crossover period (secondary analysis). SETTING AND PARTICIPANTS: Urban community in Tokyo, Japan. A total of 71 older women aged ≥70 years with MCI based on selected criteria in 689 community-dwelling women. MEASURES: Face-to-face interviews were conducted to administer the Geriatric Depression Scale (GDS) and Mini-Mental State Examination (MMSE) and collect data on medical history. Physical function measures included grip strength, knee extension strength, and usual walking speed. Blood samples were obtained to determine the levels of albumin, vitamin D, high-sensitivity C-reactive protein, and BDNF. RESULTS: Significant interactions were observed in BDNF after intervention of MFC intake in the secondary (F = 5.368, P = .024) and combined analyses (F = 4.354, P = .039) but not the primary analysis. There were no significant changes in the 3 categories of MMSE score (normal, MCI, moderate or severe cognitive impairment), GDS score, physical function, and blood indicators. CONCLUSIONS AND IMPLICATIONS: Three months of MFC ingestion had beneficial effects on BDNF levels in community-dwelling older women with MCI; however, the BDNF increases did not translate into MMSE scores. Further study into the effects of interventions on cognitive function and depression in older people with MCI is necessary.
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Factor Neurotrófico Derivado del Encéfalo/sangre , Queso , Disfunción Cognitiva/sangre , Disfunción Cognitiva/dietoterapia , Fermentación , Hongos , Anciano , Pueblo Asiatico , Biomarcadores/sangre , Proteína C-Reactiva/análisis , Queso/microbiología , Estudios Cruzados , Depresión/epidemiología , Femenino , Humanos , Vida Independiente , Japón/epidemiología , Pruebas de Estado Mental y Demencia , Fuerza Muscular , Pruebas Neuropsicológicas , Vitamina D/sangre , Velocidad al CaminarRESUMEN
Calcium-binding proteins regulate ion metabolism and the necessary signaling pathways for the maturational events of sperm. Our aim is to identify the novel calcium-binding proteins in testis. The gene EFCAB2 (GenBank NM_026626.3, NP_080902.1) was not previously examined, and its properties and exact mechanisms of action are unknown. In this study, we performed phylogenetic and structure prediction analyses of EFCAB2, which displays definitive structural features. Additionally, the distribution, localization, and calcium binding ability of mouse EFCAB2 were investigated. Results revealed extensive conservation of EFCAB2 among different eukaryotic orthologs. The constructed 3D model predicted that mouse EFCAB2 contains seven α-helices and two EF-hand motifs. The first EF-hand motif is located in N-terminal, while the second is located in C-terminal. By aligning the 3D structure of Ca2+-binding loops from EFCAB2 with calmodulin, we predicted six residues that might be involved in Ca2+ binding. The distribution of the Efcab2 mRNA, as determined by northern blotting, was detected only in the testis among mouse tissues. Native and recombinant EFCAB2 protein were detected by western blotting as one band at 20 kDa. In situ hybridization and immunohistochemical analyses showed its localization specifically in spermatogenic cells from primary spermatocytes to elongate spermatids within the seminiferous epithelium, but neither spermatogonia nor somatic cells were expressed. Moreover, EFCAB2 was specifically localized to the principal piece of cauda epididymal sperm flagellum. Furthermore, the analyses of purified recombinant EFCAB2 by Stains-all, ruthenium red staining, and by applying in vitro autoradiography assay showed that the physiological function of this protein is Ca2+ binding. These results suggested that EFCAB2 might be involved in the control of sperm flagellar movement. Altogether, here we describe about EFCAB2 as a novel calcium-binding protein in mouse testis and sperm.
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Proteínas de Unión al Calcio/fisiología , Proteínas del Citoesqueleto/fisiología , Espermatozoides/metabolismo , Testículo/metabolismo , Secuencia de Aminoácidos , Animales , Proteínas de Unión al Calcio/metabolismo , Proteínas del Citoesqueleto/metabolismo , Células HEK293 , Humanos , Masculino , Ratones , Ratones Endogámicos ICR , Espermatogénesis/genéticaRESUMEN
A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.
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The discovery of circulating tumour DNA molecules created a paradigm shift in tumour biomarkers as predictors of recurrence. Non-invasive prenatal testing (NIPT) to detect circulating cell-free foetal DNA in maternal plasma is increasingly recognised as a valuable substitute to perceive foetal copy number variation (CNV). This study aimed to determine whether the copy number detection in plasma samples using NIPT platform could be used as a prognostic biomarker in patients with gynaecological cancer. We conducted a prospective study using samples containing preoperative plasma from 100 women with gynaecological cancers. Samples were randomly rearranged and blindly sequenced using a low-coverage whole-genome sequencing plasma DNA, NIPT platform. The NIPT pipeline identified copy number alterations (CNAs) were counted in plasma as a gain or loss if they exceeded 10 Mb from the expected diploid coverage. Progression-free survival (PFS) and overall survival (OS) were analysed according to the presence of CNA in plasma using Kaplan-Meier analyses. The NIPT pipeline detected 19/100 cases of all gynaecological cancers, including 6/36 ovarian cancers, 3/11 cervical cancers, and 10/53 endometrial cancers. Patients with CNA in plasma had a significantly poorer prognosis in all stages concerning PFS and OS. Therefore, low-coverage sequencing NIPT platform could serve as a predictive marker of patient outcome.
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Ácidos Nucleicos Libres de Células/sangre , Variaciones en el Número de Copia de ADN/genética , Neoplasias de los Genitales Femeninos/sangre , Secuenciación de Nucleótidos de Alto Rendimiento , Adulto , Anciano , Anciano de 80 o más Años , Ácidos Nucleicos Libres de Células/genética , ADN Tumoral Circulante/sangre , ADN Tumoral Circulante/genética , Femenino , Pruebas Genéticas , Neoplasias de los Genitales Femeninos/clasificación , Neoplasias de los Genitales Femeninos/genética , Neoplasias de los Genitales Femeninos/patología , Humanos , Persona de Mediana Edad , Supervivencia sin Progresión , Análisis de Secuencia de ADN , Secuenciación Completa del GenomaRESUMEN
The inner cell mass of the mouse blastocyst gives rise to the pluripotent epiblast (EPI), which forms the embryo proper, and the primitive endoderm (PrE), which forms extra-embryonic yolk sac tissues. All inner cells coexpress lineage markers such as Nanog and Gata6 at embryonic day (E) 3.25, and the EPI and PrE precursor cells eventually segregate to exclusively express Nanog and Gata6, respectively. Fibroblast growth factor (FGF)-extracellular signal-regulated kinase (ERK) signalling is involved in segregation of the EPI and PrE lineages; however, the mechanism involved in Fgf4 regulation is poorly understood. Here, we identified Klf5 as an upstream repressor of Fgf4Fgf4 was markedly upregulated in Klf5 knockout (KO) embryos at E3.0, and was downregulated in embryos overexpressing Klf5 Furthermore, Klf5 KO and overexpressing blastocysts showed skewed lineage specification phenotypes, similar to FGF4-treated preimplantation embryos and Fgf4 KO embryos, respectively. Inhibitors of the FGF receptor (Fgfr) and ERK pathways reversed the skewed lineage specification of Klf5 KO blastocysts. These data demonstrate that Klf5 suppresses Fgf4-Fgfr-ERK signalling, thus preventing precocious activation of the PrE specification programme.
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Endodermo/metabolismo , Factor 4 de Crecimiento de Fibroblastos/metabolismo , Regulación del Desarrollo de la Expresión Génica , Factores de Transcripción de Tipo Kruppel/metabolismo , Animales , Blastocisto/metabolismo , Diferenciación Celular , Linaje de la Célula , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Ratones , Ratones Noqueados , Microscopía Confocal , Células Madre Pluripotentes/citología , Receptores de Factores de Crecimiento de Fibroblastos/metabolismo , Transducción de Señal , Factores de TiempoRESUMEN
BACKGROUND/AIMS: Probiotics appear to improve Helicobacter pylori-associated dyspepsia via an inhibitory effect on H. pylori; however, uncertainty exists regarding their effects in H. pylori-uninfected individuals. We evaluated the efficacy of Lactobacillus gasseri OLL2716 (L. gasseri OLL2716) on H. pylori-uninfected individuals with functional dyspepsia (FD). METHODS: A double-blind, parallel-group, placebo-controlled, randomized, controlled trial was performed. Participants were randomly assigned to ingest L. gasseri OLL2716-containing yogurt (L. gasseri OLL2716 group) or L. gasseri OLL2716-free yogurt (placebo group) for 12 weeks. Participants completed questionnaires that dealt with a global assessment as well as symptom severity. The per-protocol (PP) population was evaluated for efficacy in accordance with a plan prepared beforehand. RESULTS: Randomization was performed on 116 individuals; the PP population consisted of 106 individuals (mean age 42.8 ± 9.0). The impressions regarding the overall effect on gastric symptoms were more positive in the L. gasseri OLL2716 group compared to that in the placebo group (statistical trend; p = 0.073). The elimination rate for major FD symptoms was 17.3 and 35.3% in the placebo and L. gasseri OLL2716 groups respectively (p = 0.048). CONCLUSION: L. gasseri OLL2716 has beneficial effects on FD without H. pylori involvement.
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Dispepsia/terapia , Infecciones por Helicobacter/terapia , Lactobacillus gasseri , Probióticos/uso terapéutico , Adulto , Método Doble Ciego , Dispepsia/microbiología , Femenino , Infecciones por Helicobacter/microbiología , Helicobacter pylori/aislamiento & purificación , Humanos , Masculino , Persona de Mediana Edad , Encuestas y Cuestionarios , Resultado del Tratamiento , Yogur/microbiologíaRESUMEN
OBJECTIVES: To clarify whether ultrasonographic evaluations of fetoplacental underperfusion using umbilical artery (UmA) Doppler indices at 36 weeks' gestation can predict maternal hypertension at later gestation. METHODS: Normotensive pregnant women who underwent an ultrasound scan at 36 weeks' gestation and delivered singleton infants at term between 2012 and 2013 were prospectively enrolled. UmA Doppler and maternal blood pressure results at 36 weeks' gestation in cases with pregnancy-induced hypertension (PIH) at later gestation were compared with a control group. RESULTS: Thirty-nine and 775 cases were classified into the PIH and control group, respectively. The UmA pulsatility index (PI) and maternal systolic blood pressure (SBP) at 36 weeks' gestation were higher in the PIH group than in control group (UmA-PI: 0.88 vs. 0.80, p = 0.002; SBP: 126 mmHg vs. 112 mmHg, p < 0.001). The area under the ROC curve for the prediction of PIH by combining the UmA-PI and SBP was 0.867 (95% confidence interval (CI): 0.781, 0.954). The detection rate for PIH was 64.0% with a 10% false-positive rate. CONCLUSIONS: An increased UmA-PI at 36 weeks' gestation is associated with the occurrence of PIH at later gestation. This result may indicate the possibility to detect fetoplacental underperfusion ultrasonically.
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Hipertensión Inducida en el Embarazo/diagnóstico por imagen , Arterias Umbilicales/diagnóstico por imagen , Adulto , Estudios de Casos y Controles , Femenino , Edad Gestacional , Humanos , Recién Nacido , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Embarazo , Tercer Trimestre del Embarazo , Estudios Prospectivos , Curva ROC , Ultrasonografía Doppler de Pulso , Ultrasonografía Prenatal , Adulto JovenRESUMEN
Previously, we have identified a calcium-binding protein that is specifically expressed in spermatids and localized to the flagella of the mature sperm in mouse, so-called mCABS1. However, the physiological roles of CABS1 in the male reproductive system have not been fully elucidated yet. In the current study, we aimed to localize and clarify the role of CABS1 in porcine (pCABS1). We determined for the first time the full nucleotides sequence of pCABS1 mRNA. pCABS1 protein was detected on SDS-PAGE gel as two bands at 75 kDa and 70 kDa in adult porcine testis, whereas one band at 70 kDa in epididymal sperm. pCABS1 immunoreactivity in seminiferous tubules was detected in the elongated spermatids, and that in the epididymal sperm was found in the acrosome as well as flagellum. The immunoreactivity of pCABS1 in the acrosomai region disappeared during acrosome reaction. We also identified that pCABS1 has a transmembrane domain using computational prediction of the amino acids sequence. The treatment of porcine capacitated sperm with anti-pCABS1 antiserum significantly decreased acrosome reactions. These results suggest that pCABS1 plays an important role in controlling calcium ion signaling during the acrosome reaction.
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Proteínas de Unión al Calcio/aislamiento & purificación , Proteínas de Unión al Calcio/fisiología , Testículo/metabolismo , Acrosoma/metabolismo , Reacción Acrosómica/genética , Reacción Acrosómica/fisiología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Señalización del Calcio/genética , Señalización del Calcio/fisiología , Proteínas de Unión al Calcio/química , Proteínas de Unión al Calcio/genética , Flagelos/metabolismo , Masculino , Ratones , ARN Mensajero/genética , Espermátides/metabolismo , PorcinosRESUMEN
The gene 1700040L02Rik (GenBank accession number NM_028491, NP_082767.1) was selected by in silico screening as candidate that encodes a calcium-binding protein in sperm from a database of predicted mouse cilia-related genes. The predicted amino acid sequence revealed the presence of coiled-coil domain at the C-terminus and a CLAMP motif containing a leucine zipper domain in the middle of the protein. Assessment of a recombinant version of this protein by Stains-all and ruthenium red staining and by direct measurement of terbium binding revealed its calcium-binding activities. We therefore named this protein CABCOCO1 for calcium-binding coiled-coil protein-1. Immunohistochemical analyses showed its localization in spermatogenic cells of mouse testis. CABCOCO1 was first observed in the cytoplasm of murine spermatocytes, concentrated around centrioles of spermatids and co-localized with the centrosomal protein pericentrin. During the stage when centrosome number is reduced, CABCOCO1 relocalized to the murine sperm flagellum. On the other hand, in porcine sperm, whose proximal centriole remains intact while the distal centriole degenerates during spermiogenesis, CABCOCO1 localized both in the basal body and the flagellum. These results suggested that CABCOCO1 is involved in the control of sperm flagellar movement. Mol. Reprod. Dev. 83: 912-926, 2016 © 2016 Wiley Periodicals, Inc.
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Cuerpos Basales/metabolismo , Proteínas de Unión al Calcio/metabolismo , Centrosoma/metabolismo , Motilidad Espermática/fisiología , Cola del Espermatozoide/metabolismo , Espermátides/metabolismo , Animales , Proteínas de Unión al Calcio/genética , Leucina Zippers , Masculino , RatonesRESUMEN
The isomaltulose based liquid formula (MHN-01), suppresses postprandial plasma glucose and insulin levels in healthy persons and patients with impaired glucose tolerance (IGT) or type 2 diabetes. MHN-01 intake as a part of breakfast also suppresses glucose and insulin levels after lunch, suggesting second meal effect. The objective of this study was to investigate the effects of nutritional counseling and long-term (24 weeks) MHN-01 ingestion on biomarkers of metabolic syndrome. Forty-one subjects with criteria of metabolic syndrome participated in this study composed with the control period (0-12 week) followed by nutritional counseling and the experimental period (12-36 week) followed by 200 kcal (837 kJ) of MHN-01 or dextrin-based standard balanced liquid formula (SBF) loading as a part of breakfast. In 16 of 41 subjects became to out of criteria for liquid formula loading study during control period (unqualified group). In the unqualified group, several biomarkers were improved. In experimental period, serum HbA1c levels significantly increased in SBF group (n = 12) but did not change in MHN-01 group (n = 10). Thus, intake of 837 kJ MHN-01 as a part of breakfast may be effective for suppression of deteriorating glucose metabolism in metabolic syndrome.
RESUMEN
Maternal smoking during early pregnancy is associated with a reduced risk for preeclampsia even after smoking cessation during pregnancy. Although the pathophysiology of preeclampsia has not been established, placental growth factor (PlGF) is believed to be a key factor. The aim of this study was to assess the effect of maternal smoking on the PlGF expression in invasive trophoblasts at early gestation. We collected villous tissues from women requesting surgical termination due to non-medical reasons at 7-8 weeks of gestation. The maternal smoking status was evaluated by measuring the serum cotinine level and patients were divided into two groups: active smokers and non-smokers. After separating invasive trophoblasts from villous tissues cultured initially under 2% O2 for 24 hours, the separated invasive trophoblasts were cultured under 2% or 8% O2 for 48 hours. The expression levels of the PlGF gene in villous tissue specimens and in invasive trophoblasts cultured after the conditions were quantified using qRT-PCR. The levels of PlGF protein in the medium were quantified using an ELISA. The gene expression level of PlGF in the villi in the active-smokers was significantly higher than that of the non-smokers. In comparison of the invasive trophoblasts under normoxia and oxygenated conditions, the ratio of PlGF gene expression and protein expression under oxygenation (2% O2+8% O2 / 2% O2+2% O2) in the active-smokers were both significantly higher than in the non-smokers. Maternal smoking history appears to stimulate PlGF expression in invasive trophoblasts under oxygenated conditions. This may be one of several causes leading to the protective effect of maternal smoking on preeclampsia.
Asunto(s)
Regulación de la Expresión Génica , Proteínas Gestacionales/biosíntesis , Cese del Hábito de Fumar , Fumar/metabolismo , Trofoblastos/metabolismo , Adulto , Femenino , Humanos , Oxígeno/metabolismo , Consumo de Oxígeno , Factor de Crecimiento Placentario , Preeclampsia/etiología , Preeclampsia/metabolismo , Preeclampsia/patología , Embarazo , Fumar/efectos adversos , Trofoblastos/patologíaRESUMEN
We investigated the effect of a formula containing two different prebiotics (bifidogenic growth stimulator and galacto-oligosaccharide) and fermented milk products on intestinal microbiota and antibody responses to an influenza vaccine in enterally fed elderly in-patients. Patients were administered either formula containing prebiotics and fermented milk products (group F: n = 12, 79.9 ± 9.5 years old) or standard formula (group C: n = 12, 80.7 ± 10.1 years old) via percutaneous endoscopic gastrostomy during a 14-week intervention period. Subjects were immunized with an influenza vaccine (A/H1N1, A/H3N2, and B) at week 4 of the intervention. Blood biochemical indices, intestinal bacteria populations and antibody titers were analyzed. Bifidobacterium counts increased significantly in group F compared with group C. The enhanced antibody titers against A/H1N1 were maintained in group F for a longer period compared with group C. The titers against A/H3N2 were unchanged between both groups, and those against B were significantly lower in group F than in group C, although few subjects had seroprotective titers against A/H3N2 and B. These results suggest that administration of the formula containing prebiotics and fermented milk products may maintain antibody titers for longer periods through the improvement of intestinal microbiota.