Collagen IV significantly enhances migration and transplantation of embryonic stem cells: involvement of α2ß1 integrin-mediated actin remodeling.

Embryonic stem (ES) cell transplantation represents a potential means for the treatment of degenerative diseases and injuries. As appropriate distribution of transplanted ES cells in the host tissue is critical for successful transplantation, the exploration of efficient strategies to enhance ES cell migration is warranted. In this study we investigated ES cell migration under the influence of various extracellular matrix (ECM) proteins, which have been shown to stimulate cell migration in various cell models with unclear effects on ES cells. Using two mouse ES (mES) cell lines, ESC 26GJ9012-8-2 and ES-D3 GL, to generate embryoid bodies (EBs), we examined the migration of differentiating cells from EBs that were delivered onto culture surfaces coated with or without collagen I, collagen IV, Matrigel, fibronectin, and laminin. Among these ECM proteins, collagen IV exhibited maximal migration enhancing effect. mES cells expressed α2 and ß1 integrin subunits and the migration enhancing effect of collagen IV was prevented by RGD peptides as well as antibodies against α2 and ß1 integrins, indicating that the enhancing effect of collagen IV on cell migration was mediated by α2ß1 integrin. Furthermore, staining of actin cytoskeleton that links to integrins revealed well-developed stress fibers and long filopodia in mES cells cultured on collagen IV, and the actin-disrupting cytochalasin D abolished the collagen IV-enhanced cell migration. In addition, pretreatment of undifferentiated or differentiated mES cells with collagen IV resulted in improved engraftment and growth after transplantation into the subcutaneous tissue of nude mice. Finally, collagen IV pretreatment of osteogenically differentiated mES cells increased osteogenic differentiation-like tissue and decreased undifferentiation-like tissue in the grafts grown after transplantation. Our results demonstrated that collagen IV significantly enhanced the migration of differentiating ES cells through α2ß1 integrin-mediated actin remodeling and could promote ES cell transplantation efficiency, which may be imperative to stem cell therapy.

Small supernumerary marker chromosome originating from chromosome 10 associated with an apparently normal phenotype.

Small supernumerary marker chromosomes (sSMC) originating from chromosome 10 are rare. Only seven cases have been documented, and among those three cases were diagnosed prenatally. We reported on another prenatal diagnosis of a de novo mosaic sSMC in an apparently normal female fetus whose mother had conceived with assisted reproductive technology (ART) procedures. G-banding analysis of amniotic cells was performed. Spectral karyotyping (SKY) and fluorescence in situ hybridization (FISH) studies with chromosome 10-specific alphoid satellite DNA probe were used to identify the chromosome 10 origin of the sSMC. Further FISH study with telomeric sequence probes showed that the sSMC lacked a hybridization signal, suggesting that the marker could be a ring chromosome. FISH studies using BAC clone probes specific for the regions within 10p11.2, 10q11.1, and 10q11.2 showed that the short arm breakpoint was located between 29.8 and 30.7 Mb from the 10p telomere, and that the long arm breakpoint was located less than 43.6 Mb from the 10p telomere. The karyotype of the fetus was 47,XX,+mar. ish der(10)(SKY+ CEP 10+, CTD-2130I7+, RP11-89J23-)/46,XX. Oligonucleotide microarray-based copy number variations (CNV) analysis was also performed and showed a 6.7 Mb duplication from 10p11.2 to 10q11.2 (36.2-42.9 Mb) with Affymetrix SNP-array 6.0 genotype: arr cgh. 10p11.2q11.2(CN_519687 --> CN_541524) X 3. At the 1-year follow-up, the baby did not have any findings of the trisomy 10p syndrome. This observation provided further credence to the concept that additional chromosome material of proximal 10p11.2 may not contribute to the trisomy 10p syndrome phenotype.

Port wound closure assisted by Foley catheter: an easier way to provide fascia security.

AIM: Specific laparoscopy-related complications, such as incisional hernia (trocar hernia) or hemorrhage, are worthy of our special attention. Preventing and managing these complications safely and efficiently are important, suggesting that a definite closure of the 10-12 mm port wound after laparoscopy is needed; for this, a newer, simpler method was used. METHODS: Ninety-six patients with benign ovarian tumor warranting laparoscopic surgery were enrolled into the study. Forty-eight patients (50%) underwent a Foley catheter-assisted port wound closure (Foley group) and the remaining patients (50%) underwent a conventional port would closure (control group). The outcome was measured by comparing operative time, the amount of suture material used (difficulty of wound closure), therapeutic efficacy (hernia), postoperative complications (bleeding, hematoma, and wound pain by a self-reported six-point verbal numeric rating scale (VNRS-6)) and anesthesia use as measured by an analgesic usage score (AUS), and dissatisfaction with the cosmetic results, in both groups. RESULTS: The general characteristics of the patients were similar in both groups. There were no statistical differences in mean operative time, therapeutic efficacy, and postoperative complications between the two groups. However, the amount of extra suture material needed was significantly less in the Foley group compared to the control group (1.04 +/- 0.08 vs 1.29 +/- 0.44, P = 0.015, and 4.2 vs 25%, P = 0.02, respectively). CONCLUSION: Wound closure with the assistance of a Foley catheter offers an easy and secure way to close a 10-12 mm port wound.

Roles of estrogen and progesterone in endometrial hemodynamics and vascular endothelial growth factor production.

BACKGROUND: The endometrium becomes receptive to the embryo after sequential actions of estrogen and progesterone. The purpose of this study was to examine the effects of estrogen and progesterone on endometrial hemodynamics and on secretion of vascular endothelial growth factor (VEGF) from endometrial epithelial cells (EEC). METHODS: Six early postmenopausal women taking sequential estrogen and progestin [days 1-11: estradiol valerate (estrogen) 2 mg daily; days 12-21: estradiol valerate 2 mg plus norethisterone acetate (progestin) 1 mg daily] were recruited. Three-dimensional power Doppler angiography (3D-PDA) was performed before hormone treatment (phase 0), on days 10-11 of hormone treatment (phase E), and on days 18-20 of hormone treatment (phase E + P). Ishikawa EEC were treated with or without 17-beta-estradiol and progesterone for 24 hours, followed by determination of VEGF concentrations in the supernatants. RESULTS: The endometrial volume was significantly increased in phase E and phase E + P as compared with that in phase 0. The vascularization index, flow index, and vascularization flow index in the subendometrial region, as measured by 3D-PDA, were significantly higher in phase E + P than in phase 0, but there were no significant differences in these indices between phase 0 and phase E. While treatment of EEC with 17-beta-estradiol had little enhancing effect on VEGF production, progesterone alone or in combination with 17-beta-estradiol significantly increased VEGF secretion from EEC. CONCLUSION: Our data suggested that progesterone could stimulate VEGF secretion from EEC and subsequently increase subendometrial vascularity and blood flow.

A randomized, parallel, comparative study of the efficacy and safety of nafarelin versus danazol in the treatment of endometriosis in Taiwan.

BACKGROUND: The purpose of this study was to evaluate the efficacy and safety of nafarelin, a gonadotropin-releasing hormone (GnRH) analogue, versus danazol in the treatment of women with endometriosis in Taiwan. METHODS: Fifty-nine women with laparoscopically and pathologically confirmed endometriosis were randomized to receive nafarelin or danazol for 180 days. Efficacy was assessed from mean changes in laparoscopy score (LS) and total symptom severity score (TSSS). Adverse events (AEs) and laboratory parameters, including hematology, hepatic function, blood pressure, and lipid levels, were monitored for safety evaluations. RESULTS: All demographic and baseline factors, except body weight, were comparable between the 2 treatment groups. Both nafarelin and danazol satisfactorily resolved pelvic tenderness, induration, pelvic pain, dysmenorrhea and dyspareunia. No significant differences were noted in efficacy endpoints between nafarelin and danazol regarding LS and TSSS at 90 and 180 days of treatment. No significant difference was observed between the 2 groups regarding the overall incidence of AEs, except for laboratory-related AEs. However, nafarelin tended to have less impact than danazol on aspartate transaminase and alanine transaminase, and nafarelin was better tolerated than danazol regarding changes in lipid profiles. Both treatments had little or no effect on hematologic parameters. CONCLUSION: Nafarelin and danazol demonstrated similar clinical efficacy, but nafarelin was associated with fewer laboratory changes and a stable lipid profile, relative to danazol. Moreover, intranasally administered nafarelin is noninvasive, and may be a more comfortable and safer alternative to slow-release injectable GnRH agonists. Based on this study, we suggest that nafarelin, like other GnRH analogues, may be a treatment of choice for Taiwanese women with endometriosis. However, direct comparative studies of nafarelin with slow-release injectable GnRH agonists are now required.

Glutamic acid analogues as potent dipeptidyl peptidase IV and 8 inhibitors.

To find potent and selective inhibitors of dipeptidyl peptidase IV (DPP-IV), we synthesized a series of 2-cyanopyrrolidine with P2-site 4-substituted glutamic acid derivatives and tested their activities against DPP-IV, DPP8, and DPP-II. Analogues that incorporated a bulky substituent at the first carbon position of benzylamine or isoquinoline showed over 30-fold selectivity for DPP-IV over both DPP8 and DPP-II. From structure-activity relationship studies, we speculate that the S2 site of DPP8 might be similar to that of DPP-IV, while DPP-IV inhibitor with N-substituted glycine in the P2 site and/or with a moiety involving in hydrophobic interaction with the side chain of Phe357 might provide a better selectivity for DPP-IV over DPP8.

Magnetic resonance experience of a twin pregnancy with a normal fetus and hydatidiform mole: a case report.

A case of twin pregnancy with a coexistent hydatidiform mole after in vitro fertilization is presented. Prenatal magnetic resonance (MR) imaging at 20 gestational weeks demonstrated a normal fetus and a distinct junction between the molar tissue and myometrium suggesting no evidence of myometrial invasion. Clinically, the rare disease entity involves a high risk of maternal complications and intrauterine fetal death. The application of ultrafast MR imaging for prenatal examination provides important additional information for prenatal counseling and obstetric management.

Novel isoindoline compounds for potent and selective inhibition of prolyl dipeptidase DPP8.

DPP8 is a prolyl dipeptidase homologous to DPP-IV, which is a drug target for Type II diabetes. The biological function of DPP8 is not known. To identify potent and selective chemical compounds against DPP8, we have synthesized a series of isoquinoline and isoindoline derivatives and have tested their inhibitory activity against DPP8, DPP-IV and DPP-II. Isoindoline derivatives were found to be more potent DPP8 inhibitors than isoquinoline derivatives. Isoindoline with a 1-(4,4'-difluor-benzhydryl)-piperazine group at the P2 site was observed to be a very potent DPP8 inhibitor, having an IC(50) value of 14nM with at least a 2500-fold selectivity over either DPP-IV or DPP-II. From SAR results, we speculate that the S1 site of DPP8 may be larger than that of DPP-IV, which would allow the accommodation of larger C-terminal residues, such as isoquinoline or isoindoline.

Effect of continuous administration of conjugated estrogen plus medroxyprogesterone acetate (Premelle) in postmenopausal women in Taiwan.

BACKGROUND: To compare the metabolic effects on lipids and acceptability and safety of, and compliance with, a continuous administration of conjugated estrogen plus medroxyprogesterone acetate (Premelle) versus a placebo in non-hysterectomized postmenopausal women. METHODS: Sixty-six generally healthy, female, early post-menopausal women, from 45-60 years of age, were randomized for an administration of conjugated estrogen plus medroxyprogesterone acetate (Premelle, Premarin 0.625 mg plus medroxyprogesterone acetate 2.5 mg/tablet orally) or a placebo for 6 months. The changes in each patient's lipid profiles from baseline, the frequency of hot flushes, bleeding occurrences, and climacteric symptoms, were evaluated. Safety was monitored by means of physical examination, Papanicolau smear, transvaginal ultrasonography, and laboratory check-up. Adverse events were also recorded. RESULTS: The difference before and after treatment in serum LDL-C and total cholesterol (TC) was statistically significant in the Premelle group (LDL-C, p = 0.006, TC, p = 0.040). No statistically significant difference in the change from baseline was observed in the levels of LDL-C and TC in the placebo treatment group. There was a statistically significant change from baseline in menopausal symptoms, which were evaluated by the Greene Climacteric Scales in the Premelle group. There was no clinically significant finding in the physical examination, vital signs, laboratory data, or endometrial thickness in either treatment group. The difference in the number of patients who reported an adverse event was not statistically significant between the 2 treatment groups. CONCLUSIONS: This study demonstrated that Premelle was effective in decreasing LDL-C and total cholesterol levels, and also showed an improvement in some menopausal symptoms, such as vasomotor and sexual dysfunction symptoms. No significant bleeding was observed with Premelle, which was well tolerated in this study. The results of this study could support the use of Premelle tablets as a convenient alternative hormone therapy.

Prevalence of depressive and anxiety disorders in an assisted reproductive technique clinic.

BACKGROUND: Little is known about the prevalence of specific depressive and anxiety disorders in women before a new course of assisted reproductive technology treatment. Few studies have adopted the proper psychiatric diagnostic procedures. METHODS: All consecutive women visiting the assisted reproduction clinic of a university-affiliated medical centre, with the intention of starting a new assisted reproduction treatment course, were recruited. A psychiatrist made a diagnosis of psychiatric disorders using a structured interview, the Mini-International Neuropsychiatric Interview (MINI). RESULTS: Of a total of 112 participants, 40.2% had a psychiatric disorder. The most common diagnosis was generalized anxiety disorder (23.2%), followed by major depressive disorder (17.0%), and dysthymic disorder (9.8%). Participants with a psychiatric morbidity did not differ from those without in terms of age, education, income, or years of infertility. Women with a history of previous assisted reproduction treatment did not differ from those without in depression or anxiety. CONCLUSIONS: Depressive and anxiety disorders were highly prevalent among women who visited an assisted reproduction clinic for a new course of the treatment. Demographic features and a history of previous assisted reproduction treatment were not risk factors for these psychiatric morbidities in the assisted reproduction clinic.

A detachable porous vaginal mold facilitates reconstruction of a modified McIndoe neovagina.

OBJECTIVE: To release a new design for a detachable porous vaginal mold to facilitate reconstruction of a modified McIndoe vaginal mold. DESIGN: We constructed a detachable, porous vaginal mold with a plastic laboratory centrifuge tube with multiple holes throughout the entire tube. SETTING: Patients in a national tertiary medical center. PATIENT(S): Four patients of congenital vaginal agenesis received the modified McIndoe procedure. INTERVENTION(S): Two full-thickness skin grafts removed from the inguinal region were used to cover the detachable porous plastic vaginal mold; the mold was then inserted into the neovaginal cavity and kept in place by sutures between the mold and labia majora. The vaginal mold was removed on day 12 after the operation. All patients adhered to the follow-up instructions. MAIN OUTCOME MEASURE(S): Description of the accessibility of a neovaginal mold. RESULT(S): The vaginal mold is helpful in taking care of the vaginal wound in that it allows easy removal of wound secretion and local cleansing douches. All grafts took completely and recovered well. No detachment of the graft occurred. CONCLUSION(S): The advantages of the detachable porous vaginal mold made from a plastic centrifuge tube are that it is readily available, allows for easy wound care, and, as a fixed point, prevents graft detachment or inversion and thus may lead to a promising result for a modified McIndoe vaginal reconstruction.

Interleukin-8 can stimulate progesterone secretion from a human trophoblast cell line, BeWo.

Precise paracrine cross-talk between the embryo and the endometrium is essential for the establishment of a successful pregnancy. Previous studies have demonstrated that the expression of interleukin-8 (IL-8) in the endometrium is enhanced during the late-secretory phase and early pregnancy. Furthermore, IL-8 receptor (IL-8R) expression has been detected in trophoblast cells of the developing embryo. To clarify the roles of IL-8 in the endometrium-embryo interactions, the effects of IL-8 on hormone secretion by trophoblast cells were studied using the BeWo trophoblast cell line that retains hormone-secreting properties of normal trophoblast cells. Using reverse transcription-polymerase chain reaction, we found that IL-8R messenger ribonucleic acid (mRNA) was expressed in BeWo cells. The levels of IL-8R mRNA and protein expression in BeWo cells were similar to those in primary first-trimester trophoblast cells. Progesterone (P4) secretion of BeWo cells was comparable with that of first-trimester trophoblast cells but higher than that of third-trimester trophoblast cells. Treatment of BeWo cells with recombinant human IL-8 (rhIL-8) had no effect on cell proliferation, as determined by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Interestingly, secretion of P4, but not human chorionic gonadotropin, from cultured BeWo cells was significantly enhanced when the cells were incubated with rhIL-8. Our results demonstrate that IL-8 may play an important role in the endometrium-embryo interactions by stimulating trophoblast secretion of P4 for maintenance of a successful pregnancy.

Acute urinary retention after use of assisted reproductive technology. A report of 2 cases.

BACKGROUND: Urinary retention in women is an unusual occurrence. Various conditions can cause it, including surgery, childbirth, pelvic masses, procidentia, vulva hematoma, incarceration of a gravid uterus, urologic disease, neurogenic disease and psychiatric disorders. CASES: Two infertile women underwent assisted reproductive technology (ART) and luteal-phase support during which acute urinary retention occurred. The patients were evaluated by means of physical examination, urodynamic study and determination of serum progesterone levels. Bladder distention was found during luteal-phase support. Urodynamic studies revealed detrusor areflexia. Serial serum progesterone levels were > 100 ng/mL. CONCLUSION: Cases of acute urine retention during luteal-phase support are rarely reported. Urine retention should be considered in the differential diagnosis when abdominal pain occurs after patients undergo ART. Transient elevation of the progesterone level may be the cause of the acute urine retention.

Induction of p38 mitogen-activated protein kinase-mediated apoptosis is involved in outgrowth of trophoblast cells on endometrial epithelial cells in a model of human trophoblast-endometrial interactions.

During embryo implantation in species with hemochorial placentation, such as the mouse and human, trophoblast cells of the attached blastocyst penetrate the luminal epithelium of the endometrium before invasion into the endometrial stroma. Signs of apoptosis were demonstrated in luminal endometrial epithelial cells (EEC) adjacent to the trophoblast cells; however, the signaling mechanisms leading to apoptosis in EEC remain unclear. Because mitogen-activated protein kinases (MAPK) were shown to mediate apoptosis in several model systems and found to be activated in the uterus during decidualization, the possible involvement of MAPK during trophoblast-EEC interactions was studied. By coculturing BeWo human trophoblast spheroids with RL95-2 human EEC monolayers to mimic the blastocyst-endometrial interaction, we found that most spheroids rapidly attached to EEC monolayers and then progressively expanded, with marked dislodgment of EEC adjacent to the spreading trophoblast cells. Immunoblotting analysis showed that both p38 MAPK and extracellular signal-regulated kinase (ERK) were activated in EEC after coculture. However, only SB203580 (a p38 MAPK inhibitor), but not PD98059 (an ERK inhibitor), inhibited trophoblast outgrowth on EEC monolayers through the suppression of p38 MAPK activation in EEC. Furthermore, trophoblast expansion caused prominent EEC apoptosis at the spheroid-EEC interface, as detected by annexin V labeling and valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone (which binds activated caspases) staining, and SB203580 significantly decreased the percentage of apoptotic cells. Our results, based on a model of human trophoblast-EEC interactions, establish that trophoblast cells cause activation of p38 MAPK in EEC and, consequently, induce apoptosis and displacement of EEC, a process that may facilitate implantation.

Pregnancy following treatment of symptomatic myomas with laparoscopic bipolar coagulation of uterine vessels.

BACKGROUND: Laparoscopic bipolar coagulation of uterine vessels (LBCUV) has been employed for women with symptomatic uterine myomas, but its effect on subsequent pregnancy has not been characterized. METHODS: Four-hundred and twenty-three women entered the study between March 1999 and December 2001. Of these, 142 women (33.6%) were under the age of 40 years at the time of LBCUV, 36 of whom (36/142, 25.3%) were sexually active without contraception. In a prospective study of 142 patients (<40 years old) undergoing LBCUV for symptomatic myomas, 15 women became pregnant (17 total pregnancies) and were evaluated by physical and ultrasound examinations. RESULTS: The volume of the dominant myoma was 117.4 +/- 118.4 and 36.8 +/- 56.8 cm(3) before and after LBCUV respectively. Volume of the dominant myoma after pregnancy was 46.2 +/- 76.7 cm(3) (mean +/- SD). There was a significant difference in myoma volume before and after LBCUV (P = 0.002), but no significant difference in myoma volume when comparing post-partum size with post-LBCUV size (P = 0.269). Pregnancy outcomes included seven miscarriages in the first trimester and one premature rupture of membrane (PPROM). Although the other pregnancies were regarded as uncomplicated, only two women were delivered of normal neonates as the other seven pregnancies were terminated secondary to patient request. CONCLUSIONS: The pregnancy and term pregnancy rates in sexually active women without contraception were 41.6% (15/36) and 5.6% (2/36) respectively. Because a relatively high rate (7/17, 41.2%) of early miscarriages was observed, we recommend that this procedure be employed only for women who do not desire additional children.

Fetal meconium peritonitis in the infant of a woman with fulminant hepatitis B. A case report.

BACKGROUND: Simultaneous fulminant maternal hepatitis B infection and fetal meconium peritonitis has never been reported before in the English-language literature. CASE REPORT: Fetal meconium peritonitis was detected at 32 weeks' gestation in a 21-year-old woman suffering from fulminant hepatitis. Fulminant hepatitis B was confirmed by clinical observation and serologic examination results. The course was also complicated with preterm labor. The fetus was diagnosed with meconium peritonitis prenatally. Because of failed tocolytic treatment, the fetus was delivered vaginally. Both the mother and fetus received intensive care, and the mother recovered. In contrast, the fetus's course worsened due to progressive abdominal distension. Although exploratory laparotomy was attempted, the operation was not successful. The infant died five days after birth. CONCLUSION: Recognition of the predisposing factors in fetal meconium peritonitis and immediate referral to a tertiary medical center, where specialists are available, could help physicians determine an accurate diagnosis and might improve prognosis.

Coexistence of gonadal dysgenesis and Mullerian agenesis with two mosaic cell lines 45,X/46,X,del(X)(p22.2).

Gonadal dysgenesis and Mullerian agenesis both are common causes of primary amenorrhea. Coexistence of gonadal dysgenesis and Mullerian agenesis has been previously described as a rare event. The karyotypes, 45,XO,45,X/46,XX,45,X/46,X,dic(X),46,XX, and 46,XY, have been reported in the literature. A 22-year-old woman presented with primary amenorrhea and normal intelligence. Her physical examination confirmed the absence of breast development and axillary hair. The woman weighed 43 kg and was 150 cm tall. Scoliosis of the thoracic spine was noted on a chest X-ray film. Also, her pelvic examination revealed a vaginal introitus with a vaginal depth of 7 cm, measured by sounding. Her external genitalia were female but lacked pubic hair. The rectal examination failed to reveal a uterus. Pelvic ultrasound disclosed the absence of uterus and ovaries, and her serum gonadotropin levels were in the menopausal range (FSH, 118.59 IU/L; LH, 38.94 IU/L). Estradiol was less than 10 pg/ml. Two mosaic cell lines, 45,X (50%) and 46,X,del(X)(p22.2X50%), were found in the chromosomal study. Laparoscopic evaluation confirmed the absence of uterus and ovaries with normal fallopian tubes. Coexistence of gonadal dysgenesis and Mullerian agenesis is a rare event. The two mosaic cell lines 45,X/46,X,del(X)(p22.2) in this combination have not been reported before. In patients with this condition, estrogen will initiate and sustain maturation and function of secondary sexual characteristics, and lifelong hormone therapy will protect against osteoporosis and cardiovascular disease.