OBJECTIVE: We previously described that Diva is highly expressed in matured metaphase II (MII) oocytes compared to immature germinal vesicle (GV) oocytes in mouse.1 We report here that the expression of Diva transcript as well as protein is oocyte-specific. To elucidate its physiological role in oocyte, the binding partner(s) of Diva has been identified by using immunoprecipitation (IP) followed by Mass Spectrometry. METHODS: NIH/3T3 cells were transiently transfected for 24 h with either empty vector for control or FLAG-tagged mouse Diva construct, and IP was performed with anti-FLAG antibody. The immuno-isolated complexes were resolved by SDS-PAGE on a 12% gel followed by Coomassie Blue staining. For in-gel digestion, 15 bands of interest were excised manually and digested with trypsin. All mass spectra were acquired at a positive reflector mode by a 4700 Proteomics Analyzer (Applied Biosystems, Framingham, MA). Proteins were identified by searching the NCBI nonredundant database using MASCOT Peptide Mass Fingerprint software (Matrixscience, London). RESULTS: Diva-associated complexes were formed in FLAG-tagged mouse Diva-overexpressed NIH/3T3 cells via IP using anti-FLAG-conjugated beads. Among the excised 15 bands, actin and actin-binding proteins such as tropomyosin, tropomodulin 3, and alpha-actinin were identified. Binding between Diva and actin or tropomyosin was confirmed by IP followed by Western blot analysis. Both bindings were also detected endogenously in mouse ovaries, indicating that Diva works with actin and tropomyosin. CONCLUSIONS: This is the first report that immuno-isolated Diva-associated complexes are related to actin filament of the cytoskeletal system. When we consider the association of Diva with actin and tropomyosin, oocyte-specific Diva may play a role in modulating the cytoskeletal system during oocyte maturation.
Actin Cytoskeleton ; Actinin ; Actins ; Animals ; Blotting, Western ; Dermatoglyphics ; Digestion ; Electrophoresis, Polyacrylamide Gel ; Female ; Immunoprecipitation ; Mass Spectrometry* ; Metaphase ; Mice ; Microfilament Proteins ; Oocytes ; Ovary ; Proteomics ; Tropomodulin ; Tropomyosin ; Trypsin

No abstract available.
HIV-1* ; Humans* ; Tumor Necrosis Factor-alpha*

BACKGROUND: Thromboembolism is the major complication in patients with the insertion of cardiac prosthetic valves. The purpose of this study is evaluate the risk factors about the formation of left atrial thrombi after cardiac valve replacement. METHOD: Transesophageal(TEE) and transthoracic echocardiography(TTE) were done to evaluate postoperative cardiac condition in 98 patients with cardiac prosthetic valves from Jan. 1991 to Oct 1991. Several clinical and echocardiographic parameters were analyzied to evaluate the relationship with the formation of left atrial thrombi. RESULT: In univariate analysis, important factors related to the formation of left atrial thrombi are type of operation (p=0.027), postoperative left ventricular function(p=0.003), preoperative(p=0.037) and postoperative systolic ventricular size(p=0.024). However, in multivariate analysis postoperative left ventricular size(p=0.017), presence of previous thrombi(p=0.014), preoperative left atrial size(p=0.014) and postoperative left atrial size(p=0.014) are significant factors. CONCLUSION: Patients with high risk and low risk for the formation of left atrial thrombi after valve replacement can be identified by readily available clinical and echocardiographic variables.
Echocardiography ; Heart Valves ; Humans ; Multivariate Analysis ; Risk Factors* ; Thromboembolism

Visceral afferent nerve fibers containing substance P or calcitonin gene-related peptide (CGRP) are distributed in the bladder wall, and are known to be stimulated by and then desensitized by capsaicin. Recently, there have been some reports on the effectiveness of intravesical capsaicin administration for the treatment of hypersensitive lower urinary tract disorder or neurogenic bladder. In this study, the effects of intravesical capsaicin on the substance P or CGRP immunoreactivities in the spinal dorsal horn were investigated and the mechanism of capsaicin treatment for bladder disorders was revealed. After intravesical administration of capsaicin, the substance P and CGRP immunoreactive areas were measured at the dorsal horn of L4 and S1 spinal cord. Before capsaicin treatment, the substance P immuno- reactive area was 2.61+/-0.78 x 105 mm2 in L4 and 1.66+/-0.49 x 105 mm2 in S1. The substance P immunoreactivity was markedly reduced 1~2 weeks after capsaicin treatment in both L4 and S1 spinal cord. The CGRP immunoreactive area was 1.74+/-0.52 x 105 mm2 in L4 and 1.14+/-0.69 x 105 mm2 in S1, but was not reduced after capsaicin treatment. Therefore, capsaicin administered intravesically desensitizes nerve fibers containing substance P and consequently suppresses pain and voiding reflex.
Administration, Intravesical ; Animals ; Calcitonin Gene-Related Peptide ; Capsaicin* ; Horns* ; Nerve Fibers ; Rats* ; Reflex ; Spinal Cord ; Substance P ; Urinary Bladder ; Urinary Bladder, Neurogenic ; Urinary Tract ; Visceral Afferents

Calcitonin gene-related peptide (CGRP), a well-known neuropeptide in primary sensory neurons carrying noci-ceptive information, includes two different peptides of similar structure, the alpha- and beta-CGRP. The distribution of these two peptides in the central nervous system is known to be similar and no functional differences have been reported. The aim of this study is to investigated the changes of alpha- and beta-CGRP expression following efferent or afferent disconnection of anterior horn cells in the rat spinal cord. One week after ventral rhizotomy (left L4-6), dorsal rhizotomy (left L4-6) or spinal cord transection (at lower thoracic level), the animals were sacrified and the L5 segments of the spinal cord were excised to perform immunohistochemistry and in situ hybridization histochemistry. In the control group, 4.45+/-1.51 anterior horn cells showed CGRP immunoreactivity per tissue section in one side. After ventral rhizotomy, the number of CGRP immunoreactive neurons increased to 9.12+/-2.52 at the ipsilateral ventral horn. After dorsal rhizotomy, CGRP immunoreactive neurons increased to 7.29+/-3.69 at the ipsilateral ventral horn and 6.26+/-1.53 at the contralateral ventral horn. In cases of spinal cord transection, almost all the anterior horn cells lost CGRP immunoreactivity in both sides. Neurons expressing alpha- or beta-CGRP mRNA could be distinguished by in situ hybridization histochemistry. In the control group, anterior horn cells expressing alpha-CGRP mRNA numbered 4.16+/-1.32 per section and beta-CGRP cells numbered 5.50+/-1.38. After ventral rhizotomy, the number of cells expressing alpha-CGRP mRNA increased to 10.07+/-2.86 in the ipsilateral side without any changes in beta-CGRP mRNA expression. After dorsal rhizotomy, no significant changes in alpha-CGRP mRNA expression were detected, but the number of cells expressing beta-CGRP mRNA increased to 7.45+/-2.04 in the ipsilateral side and to 7.02+/-1.38 in the contralateral side. In cases of spinal cord transection, the anterior horn cells lost alpha- and beta-CGRP mRNA signals almost completely in both sides. These results showed that alpha-CGRP expression increased in axotomized anterior horn cells of the spinal cord, and that beta-CGRP expression increased in anterior horn cells which had lost their afferent input through the primary sensory neurons. These findings provide evidence showing the functional difference of the two peptides in anterior horn cells of the spinal cord.
Animals ; Anterior Horn Cells ; Calcitonin Gene-Related Peptide ; Central Nervous System ; Horns ; Immunohistochemistry ; In Situ Hybridization ; Neurons ; Neuropeptides ; Peptides ; Rats* ; Rhizotomy* ; RNA, Messenger ; Sensory Receptor Cells ; Spinal Cord Injuries* ; Spinal Cord*

PURPOSE: Chromosomal abnormalities of abortuses have been used to investigate common etiologies of spontaneous abortion, but the frequencies and types of spontaneous abortions have demonstrated considerable variation among different countries and races. MATERIALS AND METHODS: A cytogenetic analysis of 75 abortuses was performed at GenDix, Inc. from January 2006 to December 2007. RESULTS: The frequency of chromosome abnormalities in abortuses was 32.0% (24/75 cases). Among the chromosomal abnormalities, trisomy was present in 62.5% (15/24 cases) of cases and the most frequent trisomy was trisomy 21 with an occurrence rate of 26.6% (4/15 cases). The following was trisomy 22 (3/15 cases) and trisomy 20 (2/15 cases). The average maternal age for abnormal karyotypes was 34.3+/-3.3. CONCLUSION: Cytogenetic analysis of abortus is important for diagnosis and genetic counseling of patients with spontaneous abortion.
Abnormal Karyotype ; Abortion, Spontaneous ; Chromosome Aberrations ; Chromosomes, Human, Pair 20 ; Chromosomes, Human, Pair 22 ; Cytogenetic Analysis ; Cytogenetics ; Down Syndrome ; Female ; Genetic Counseling ; Humans ; Karyotype ; Maternal Age ; Mosaicism ; Pregnancy ; Trisomy

OBJECTIVES: Previously, we sought to compile a list of genes expressed during early folliculogenesis by using cDNA microarray to investigate follicular gene expression and changes during primordialprimary follicle transition and development of secondary follicles (Yoon et al., 2005). Among those genes, a group of genes related to the cell size growth was characterized during the ovarian development in the present study. METHODS: We determined ovarian expression pattern of six genes related to the cell size growth (cyr61, emp1, fhl1, socs2, wig1 and wisp1) and extended into CCN family (connective tissue growth factor/cysteine-rich 61/nephroblastoma-overexpressed), ctgf, nov, wisp2, wisp3, including cyr61 and wisp1 genes. Expression of mRNA and protein according to the ovarian developmental stage was evaluated by in situ hybridization, and/or semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), and immunohistochemistry, respectively. RESULTS: Among 6 genes related to the cell size growth, cyr61 and wisp1 mRNA was detected only in oocytes in the postnatal day5 mouse ovaries. cyr61 mRNA expression was limited to the nucleolus of oocytes, while wisp1 was expressed in the cytoplasm and nucleolus of oocytes, except nucleus. cyr61 mRNA expression, however, was found in granulosa cells from secondary follicles. The rest 4 genes in the cell size growth group were detected in oocytes, granulosa and theca cells. Cyr61 and Wisp1 proteins were expressed in the oocyte cytoplasm from primordial follicle stage. Especially, Cyr61 protein was detected in pre-granulosa cells, Wisp1 protein was not. By using RT-PCR, we evaluated and decided that Cyr61 protein is produced by their own mRNA in pre-granulosa cells that was not detected by in situ hybridization. cyr61 and wisp1 genes are happen to be the CCN family members. The other members of CCN family were also studied, but their expression was detected in oocytes, granulose and theca cells. CONCLUSIONS: We firstly characterized the ovarian expression of genes related to the cell size growth and CCN family according to the early folliculogenesis. Cyr61 protein expression in the pre-granulosa cells is profound in meaning. Further functional analysis for cyr61 in early folliculogenesis is under investigation.
Animals ; Cell Enlargement* ; Cell Size* ; Cysteine-Rich Protein 61 ; Cytoplasm ; Female ; Gene Expression ; Genes, vif ; Granulosa Cells ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Mice* ; Oligonucleotide Array Sequence Analysis ; Oocytes ; Ovary ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Messenger ; Theca Cells

INTRODUCTION There are three factors for successful implantation. These are embryo quality, uterine receptivity, and synchronization between embryonic and endometrial development. Despite remarkable progress in investigating embryos in human IVF, there has been slow progress in exploring the implantation process. It may be due to two reasons as follow. First, it is difficult to directly investigate the mechanism of implantation in the human, because of ethical considerations. Second, there is no sensitive and widely accepted marker for assessing endometrial development. Since the finding of a novel standard for dating endometrial biopsy by Noyes et. al.,. in 1950, there have been many attempts to identify suitable markers for uterine receptivity. Those include ultrasonographic changes (Ueno et.al., 1991; Grunfeld et al.,1991), three dimensional morphological changes of the endometrium such as pinopode formation (Market or alphaf., 1987; Mantel or alphaf., 1991; Nikas et al., 1995; Psychoyos & Nikas, 1994), integrin expression (Ilesanmi et al., 1993; Lessey et.al., 1992; Lessey, 1994), and measurement of endometrial proteins (Hell, 1986;Fay & Crudzinskas, 1991). Investigations in the rat (cartel et al., 1991)and human (cartel et al., 1987; Nikas et al., 1995; Psychoyos & Nikas, 1994) suggested the presence of pinopodes as a marker for the receptive phase.4 chronological barrier in uterine receptivity could be one of the major factors limiting IVF pregnancy rates. If we were able to manage the 'implantation window' we may be able to improve implantation and pregnancy rates in the human IVF program. In 1987, Martel et al., found early appearance of pinopodes in stimulated cycles for IVF compared to natural cycles in humans (Marcel et al., 1987). This effect was found in patients stimulated with clomephene citrate/hMG/hCG. The purpose of the present study was to evaluate the endometrial development in IVF patients stimulated with either by FSH/hMG/hCG or with GnRH agonist down regulation.
Animals ; Biopsy ; Down-Regulation ; Embryonic Structures ; Endometrium ; Female ; Gonadotropin-Releasing Hormone ; Humans* ; Oocyte Retrieval* ; Oocytes* ; Pregnancy Rate ; Rats

PURPOSE: Leptin is a highly hydrophilic 16-kDa protein which is produced in the adipose tissue and participates in the regulation of food intake and energy expenditure. The aim of the present study was to examine the relation between umbilical cord blood leptin concentration and intrauterine growth. METHODS: Ninety-seven full-term newborn infants who were born in Yeungnam University Hospital from July to August 1998 were included in the study. They were divided into 3 groups related to birth weight : appropriate for gestational age(AGA) group(n=73), large for gestational age(LGA) group(n=17), small for gestational age(SGA) group(n=7). Birth weight, head circumference, mid-arm circumference, mid-arm circumference to head circumference ratio, Ponderal index, and BMI were measured at birth. Maternal body weight and placental weight were measured. Leptin concentrations of cord blood and maternal serum were measured by a RIA method, and testosterone, estradiol, insulin, c-peptide, glucose, white blood cell, hemoglobin, platelet count of cord blood were also measured. RESULTS: Leptin concentration in cord blood was positively correlated to birth weight and body length. Leptin concentrations(microgram/L) in cord blood were significantly different among groups(10.1+/-1.1 in LGA group, 8.7+/-0.9 in AGA group, 1.7+/-0.1 in SGA group). There was a statistically significant difference in leptin concentration of cord blood between female and male infants(11.6+/-1.9, versus 6.7+/-0.9). There was no significant correlations between leptin concentration of cord blood and placental weights or maternal leptin concentration. Therefore leptin concentration of cord blood can not inflect maternal leptin concentration but intrauterine fetal growth. CONCLUSION: Leptin in cord blood might originate mainly from fetal adipose tissue rather than the placenta, and may be related to fetal growth.
Adipose Tissue ; Birth Weight ; Body Weight ; C-Peptide ; Eating ; Energy Metabolism ; Estradiol ; Female ; Fetal Blood* ; Fetal Development* ; Glucose ; Head ; Humans ; Infant, Newborn ; Insulin ; Leptin* ; Leukocytes ; Male ; Parturition ; Placenta ; Platelet Count ; Testosterone ; Weights and Measures

OBJECTIVE: This study was carried out to compare the effects of the stepwise exposure treatments on the morphological normality, fertilization and blastocyst formation rate of the frozen-thawed mouse mature oocytes by vitrification or ultra-rapid freezing and to use as a fundamental data for the cryopreservation of human oocytes. MATERIALS AND METHODS: The morphological normality and fertilization rates of the vitrified and ultra-rapid frozen mouse mature oocytes after three-stepwise exposure treatments (1step, 3step and 5step) were observed. After choosing the 3step exposure treatment groups, we observed the morphological normality and fertilization, blastocyst formation rate vitrified and ultra-rapid frozen mouse mature oocytes. RESULTS: The morphological normality and fertilization rates of the vitrified mouse mature oocytes after three-stepwise exposure treatments (1step, 3step and 5step) were 75%, 85%, 88% and 58%, 61%, 54% respectively. There were no significant differences among treatments (p>0.05). The morphological normality and fertilization rates of the control was 92% and 65%. There were no significant differences in fertilization rate among control and treatments (p>0.05). The morphological normality and fertilization rates of the ultra-rapid frozen mouse mature oocytes after three-stepwise exposure treatments (1step, 3step and 5step) were 83%, 83%, 84% and 75%, 63%, 56% respectively. There were no significant differences among treatments (p>0.05). The morphological normality and fertilization rate of the control was 95% and 67%. There were no significant differences among control and treatments (p>0.05). The morphological normality and fertilization rate of the vitrified or ultra-rapid frozen mouse mature oocytes after 3step exposure treatment were 69% and 75%, respectively. The blastocyst formation rate was 60% and 57%. The results did not differ significantly between vitrification and ultra-rapid freezing (p>0.05). CONCLUSION: As known in the above results, there were no significant differences in the fertilization and blastocyst formation rate of the frozen-thawed mouse mature oocytes by vitrification or ultra-rapid freezing among the control and treatments. It is suggested that vitrification and ultra-rapid freezing method were effective for the cryopreservation of mouse mature oocytes.
Animals ; Blastocyst ; Cryopreservation ; Fertilization ; Freezing* ; Humans ; Mice* ; Oocytes* ; Vitrification*