Patients with KCNJ11-related diabetes frequently have neuropsychological impairments compared with sibling controls.

AIMS: KCNJ11-related diabetes is the most common form of permanent neonatal diabetes and has been associated with a spectrum of neurodevelopmental problems. We compared neurodevelopmental outcomes in patients with KCNJ11 mutations and their sibling controls. METHODS: Through our Monogenic Diabetes Registry (http://monogenicdiabetes.uchicago.edu/), we evaluated 23 patients with KCNJ11 mutations with (n = 9) and without (n = 14) global developmental delay successfully treated with sulfonylurea and 20 healthy sibling controls, using a battery of targeted neuropsychological and behavioural assessments with scaled scores that are comparable across a wide range of ages. RESULTS: Patients with KCNJ11-related diabetes without global developmental delay had significant differences compared with sibling controls on a range of assessments including IQ, measures of academic achievement and executive function. KCNJ11 patients with global delay exhibited significant differences in behavioural symptoms with a tendency to avoid social contact and displayed a reduced ability to adapt to new circumstances. Parents reported more immature behaviour, gross mood swings, bizarre thoughts, other unusual and severe behaviours, and there were also significant deficits in all subdomains of daily living skills. CONCLUSIONS: This series represents the largest and most comprehensive study of neuropsychological and behavioural dysfunction of individuals with KCNJ11 diabetes and is the first to compare outcome with sibling controls. Our data demonstrate the variety of neurodevelopmental problems seen in those with KCNJ11 mutations, even in those without recognized global developmental delays. These data can be used to counsel families and guide structured neurodevelopmental assessments and treatments based on the initial genetic diagnosis in patients with neonatal diabetes.

Successful transvaginal aspiration of interstitial pregnancy after failed methotrexate treatment.

Interstitial pregnancy is an uncommon condition that is challenging, not only in making an accurate diagnosis, but also in the choice of treatment. Systemic methotrexate (MTX) treatment has been favored to prevent scarring of the uterus. Nevertheless, surgery is generally indicated when this treatment fails. Transvaginal aspiration of the gestational tissue has been proposed as an alternative to surgery. The authors present a case of interstitial pregnancy in which the patient failed to respond to multidose MTX treatment and was successfully treated with transvaginal sonography-guided transvaginal aspiration of the gestational tissue, thereby bypassing the risk associated with undergoing major surgery. Transvaginal aspiration of conceptive tissue may be a novel treatment for patients with unruptured interstitial pregnancy.

Effect of cetrorelix dose on premature LH surge during ovarian stimulation.

Previous studies have shown that ovarian stimulation with clomiphene citrate (CC), human menopausal gonadotrophin (HMG), and multiple-dose gonadotrophin-releasing hormone (GnRH) antagonist is associated with a high rate of premature LH surge. This study assessed whether administration of the GnRH antagonist cetrorelix at an incremental dose or at a high dose (0.5mg) from the start could prevent premature LH surge. Couples with male factor or unexplained infertility who were going to undergo intrauterine insemination were randomized into two stimulation protocols. All women were stimulated with CC and HMG. In protocol A, cetrorelix was given at 0.25 mg per day when the leading follicles reached 14 mm, and increased to 0.5 mg when the leading follicles were 16 mm. With protocol B, cetrorelix was given at 0.5 mg per day when the leading follicles reached 14 mm. The primary outcome measure was the incidence of premature LH surge. Premature LH surge occurred in 21.6% of patients undergoing protocol A, and in 18.9% of patients undergoing protocol B. Cetrorelix at incremental dose or at 0.5 mg per day does not prevent premature LH surges associated with the CC/HMG/multiple-dose cetrorelix stimulation protocol.

Induction of topoisomerase II gene expression in human lymphocytes upon phytohemagglutinin stimulation.

The carboxyl-terminal one-third of human topoisomerase II polypeptide expressed in Escherichia coli was used as antigen to generate polyclonal antibodies in rabbits. With the use of antiserum, DNA topoisomerase II levels of phytohemagglutinin-stimulated human lymphocytes were measured by immunoblotting. Our results showed that the increase in intracellular topoisomerase II level paralleled the entry of cells into proliferation. We also found that the increase in the topoisomerase II level resulted from an increase in the amount of topoisomerase II mRNA. The time course study indicated that the appearance of topoisomerase II mRNA was first observed at 36 h after phytohemagglutinin stimulation. The maximal level of topoisomerase II mRNA was seen at 45 h after stimulation. The same RNA blot was rehybridized with a thymidine kinase probe. The maximal level of thymidine kinase mRNA was observed at 39 h after phytohemagglutinin stimulation. In a comparison of the time course of topoisomerase II gene expression with that of [3H]thymidine incorporation and thymidine kinase gene expression, it was found that the expression of the topoisomerase II gene was later than the onset of DNA replication. Thus, this study suggests that topoisomerase I, which is constantly expressed throughout the cell cycle, might participate in the initiation of DNA replication, while topoisomerase II is involved in solving the DNA topological problems accompanying DNA strand separation during DNA replication.

Studies of topoisomerase-specific antitumor drugs in human lymphocytes using rabbit antisera against recombinant human topoisomerase II polypeptide.

A complementary DNA fragment of the human DNA topoisomerase II gene was cloned into a T7 expression vector and overproduced in Escherichia coli. Rabbit polyclonal antibodies were raised against the recombinant topoisomerase II polypeptide which corresponds to the C-terminal one-third of human topoisomerase II polypeptide. Using the antiserum, DNA topoisomerase II levels were measured by immunoblotting human lymphocytes following phytohemagglutinin (PHA) stimulation. Our results showed that the intracellular topoisomerase II but not the topoisomerase I level increased in parallel with the entry of cells into proliferation. At least a 100-fold increase in topoisomerase II was observed at 50 h after PHA stimulation. As topoisomerase II levels increased upon PHA stimulation, DNA damage induced by teniposide (VM26) increased in parallel, as measured by both DNA synthesis inhibition and chromosomal aberrations. However, the damage induced by camptothecin also increased upon PHA stimulation, while the level of topoisomerase I remained relatively constant. Our results suggest that, in addition to cellular contents of topoisomerases, the state of cell proliferation is another important determinant of drug action.

Isolation of human KB cell lines resistant to epidermal growth factor-Pseudomonas exotoxin conjugates.

Mutants of the human KB carcinoma cell line resistant to a cytotoxic conjugate of epidermal growth factor (EGF) and Pseudomonas exotoxin (PE) were selected. EGF-PE and the drug verapamil, which enhanced EGF-PE cytotoxicity, were used in the selection process. These mutants also showed some cross-resistance to PE. All of the EGF-PE resistant variants displayed lower levels of 125I-EGF binding, 20-50% of parental KB levels, without altered affinity for EGF and grew at a slower rate than the parental cell line KB-3-1. These results indicate that EGF-PE resistant KB cells have a complex phenotype which includes a reduction in the number of EGF receptors and reduced sensitivity to unconjugated PE. Resistance to toxin-conjugates, although pleiotropic, is specific and does not lead to resistance to multiple other anticancer drugs, nor are independently selected multidrug resistant KB lines resistant to PE. These results argue that protocols for cancer treatment could effectively use specifically designed cytotoxic toxin conjugates as an adjunct to conventional chemotherapy.

Pregnancy after immature oocyte donation and intracytoplasmic sperm injection.

OBJECTIVE: To report a case of pregnancy from in vitro-matured primary oocytes fertilized by ICSI. The pregnancy occurred in a woman who was in an oocyte donation program; the woman's husband had normal sperm parameters. DESIGN: Case report. SETTING: Private general hospital affiliated with a university hospital. PATIENT(S): A recipient with premature ovarian failure, a recipient's husband with normal sperm, and a pregnant woman who donated her oocytes. INTERVENTION(S): Aspiration of immature oocytes during cesarean section, in vitro culture for maturation, ICSI of matured oocytes, coculture of fertilized oocytes. MAIN OUTCOME MEASURE(S): Fertilization of oocytes by ICSI, and cleavage of embryos by Vero cell coculture. RESULT(S): Two of seven immature oocytes became metaphase II oocytes, and both were fertilized by ICSI. The two zygotes were cocultured on Vero cells to become grade 1 two-cell embryos. Pregnancy was obtained after transfer. CONCLUSION(S): More studies are necessary to clarify whether ICSI can increase the fertilization rate of in vitro-matured primary oocytes, and to clarify the role of coculture in fertilization.

The role of blocking factors and antipaternal lymphocytotoxic antibodies in the success of pregnancy in patients with recurrent spontaneous abortion.

OBJECTIVE: To elucidate the role of mixed lymphocyte reaction blocking factors (BF) and complement-dependent antipaternal lymphocytotoxic antibodies on the outcome of pregnancy in unexplained recurrent spontaneous aborters. DESIGN: A controlled study of immunotherapy in which the treated group was immunized with the husband's or a third party donor's lymphocytes and the control group received autologous lymphocytes. SETTING: Tertiary care institution. PATIENTS: Forty-three patients in the control group and 48 patients in the treated group. INTERVENTION: The before and after immunization levels of BF and antipaternal lymphocytotoxic antibodies were measured. MAIN OUTCOME MEASURES: The existence or changing pattern of BF and antipaternal lymphocytotoxic antibodies levels before and after immunization had no influence on the pregnancy outcome in either group of patients. CONCLUSION: Neither BF nor antipaternal lymphocytotoxic antibodies is essential for successful pregnancy. They probably reflect the immunological response of the mother to exposure to fetal antigens.

Increase in the expression of killer cell inhibitory receptors on peritoneal natural killer cells in women with endometriosis.

OBJECTIVE: Malfunction of peritoneal natural killer cells (NK) may result in endometriosis. The present study was designed to determine whether the decrease in NK cytotoxicity occurs at early and advanced stages of endometriosis and is due to the increase in the NK inhibition receptors. DESIGN: A case control study. SETTING: A tertiary-care infertility center . PATIENT(S): A total of 44 women (controls, n = 11; women with early-stage endometriosis, n = 11; and women with advanced-stage endometriosis, n = 22) were included in this study. INTERVENTION(S): Laparoscopic examination. MAIN OUTCOME MEASURE(S): NK cytotoxicity was determined by assay of (51)Cr release against K562 cells, and the expression of killer cell inhibitory receptors (KIR, including NKB1, GL183, and EB6) in NK cells was examined by flow cytometry. RESULT(S): Women with endometriosis showed a decrease in peritoneal NK cytotoxicities against K562 at early and advanced stages of endometriosis. The expression of KIR (NKB1 and EB6) was significantly elevated in the peritoneal NK cells of women with advanced-stage endometriosis compared with controls. KIR (NKB1) was also significantly increased in peritoneal NK cells of women with advanced-stage endometriosis, compared with those of women with early-stage endometriosis. CONCLUSION(S): The results of this study suggest that the decrease in peritoneal NK cytotoxicities against K562 is observed and that this disease may be partially due to the increased expression of KIR on these NK cells.

Percutaneous epididymal sperm aspiration in psychogenic anejaculation during IVF. A report of two cases.

BACKGROUND: Psychogenic anejaculation on the day of oocyte retrieval is uncommon in in vitro fertilization (IVF) programs. Before the use of intracytoplasmic sperm injection (ICSI), either the cycle had to be canceled or a sperm donor recruited. CASES: Two couples were enrolled for IVF. On the day of oocyte retrieval, the male partners developed psychogenic anejaculation. CONCLUSION: In cases of psychogenic anejaculation during IVF, percutaneous epididymal sperm aspiration can be utilized to obtain sperm for ICSI.

Methotrexate for cesarean scar pregnancy after in vitro fertilization and embryo transfer. A case report.

BACKGROUND: Cesarean scar pregnancy is an exceedingly rare occurrence. We present the first case of cesarean scar pregnancy following in vitro fertilization-embryo transfer (IVF-ET). CASE: A 40-year-old woman with a history of a previous cesarean section presented with five years of unexplained infertility. The patient complained of abdominal pain 16 days after embryo transfer. Ultrasonography revealed a gestational sac with cardiac activity located outside the lower segment of the uterus. Dilatation and curettage was performed due to misdiagnosis of inevitable abortion. Two weeks later, repeated sonography demonstrated a sacculus, 4.07 x 4.07 cm, within the uterine isthmus with only 7.1 mm of thickness separating the sac from the urinary bladder. Normal cervical length without ballooning was noted. Cesarean scar pregnancy was diagnosed. Local injection of methotrexate (MTX) under ultrasound guidance was performed. Plasma beta-hCG levels declined from 23,328 to 8 mlU/mL within two months. CONCLUSION: For women with cesarean scar pregnancy who desire fertility, conservative treatment using MTX is an excellent choice.

Prenatal diagnosis of meconium peritonitis in a twin pregnancy after intracytoplasmic sperm injection. A case report.

BACKGROUND: Meconium peritonitis occurring in pregnancies following artificial reproductive techniques (ART) is rare. We report the first case of meconium peritonitis following intracytoplasmic sperm injection (ICSI). CASE: A 37-year-old woman attended our in vitro fertilization (IVF) program because her husband suffered from hypospermatogenetic azoospermia due to cancer surgery and radiotherapy. The patient achieved a twin pregnancy through ICSI from testicular sperm extraction at our IVF center. Meconium peritonitis, fetal ascites, polyhydramnios, bowel dilatation, hydrocele and intraabdominal calcification were noted in one of the twins on ultrasound at 30 weeks' gestation. Cesarean section due to breech presentation in labor was performed at 36 weeks' gestation. A normal female and male infant with a distended abdomen were delivered. Emergency laparotomy was performed on the male twin because of dyspnea. A 0.2-cm perforation was found in the terminal ileum. Ileotomy was performed and closed after 27 days. CONCLUSION: Prenatal diagnosis of meconium peritonitis is possible through careful ultrasonographic examination, and early surgical intervention and intensive postoperative support are required to improve the prognosis.

Pregnancy after intentional cryopreservation of percutaneous sperm aspiration specimens and intracytoplasmic sperm injection. A report of two cases.

BACKGROUND: Microsurgical epididymal sperm aspiration (MESA) and percutaneous epididymal sperm aspiration (PESA) are two methods of obtaining spermatozoa from patients with irreparable obstructive azoospermia. Intentionally using frozen-thawed spermatozoa obtained from MESA had been reported to be successful. With minimal invasiveness, intentionally cryopreserved PESA specimens and intracytoplasmic sperm injection (ICSI) were carried out in two cases. CASES: Two cases of irreparable obstructive azoospermia received PESA, and the spermatozoa were cryopreserved intentionally. Successful ICSI was performed later, utilizing frozen-thawed spermatozoa. CONCLUSION: PESA and ICSI are promising methods for these patients. The major advantages are minimal invasiveness and flexibility for further treatment.

Serum copper and zinc levels in patients with cervical cancer.

The serum copper (SCL) and zinc (SZL) levels were measured in 99 patients with cervical cancer and 50 patients with uterine myoma as controls. The mean SCL in the control group was 109.4 +/- 17.4 micrograms/ml as compared to 117.1 +/- 14.6 micrograms/dl and was not significant (NS) in 17 carcinoma in situ (CIS) patients, 142.3 +/- 14.2 micrograms/dl in 30 stage I patients (p less than 0.001), 159.0 +/- 16.6 micrograms/dl in 22 stage II patients (p less than 0.001), 171.6 +/- 25.7 micrograms/dl in 10 stage III or IV patients (p less than 0.001), and 166.2 +/- 32.2 micrograms/dl in 20 recurrent patients (p less than 0.001). The SCL returned to control level 2 weeks after surgical treatment for the stage I and II patients (mean 110.6 +/- 19.6 and 108.7 +/- 20.4 micrograms/dl, respectively, p less than 0.001). The SZL was 97.2 +/- 15.8 micrograms/dl in control patients and only showed a significant decrease in stage III or IV and recurrent patients (67.2 +/- 16.6 and 70.4 +/- 17.2 micrograms/dl, respectively). Concerning the copper/zinc ratio, the control group was 1.13 +/- 0.07 as compared to 1.17 +/- 0.07 in CIS (p = 0.06), 1.51 +/- 0.24 in stage I (p less than 0.001), 1.85 +/- 0.37 in stage II (p less than 0.001), 2.66 +/- 0.61 in stage III or IV (p less than 0.001), and 2.50 +/- 0.75 in recurrent patients (p less than 0.001). Taking mean +/- 2.5 SD of the control values as cut off points, the percentages of the recurrent patients with abnormal SCL, SZL, and a Cu/Zn ratio were 65, 30 and 90%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Placenta increta in the second trimester of pregnancy: report of a case.

A rare case of placenta increta in the second trimester of pregnancy is reported. The patient was at 15 weeks gestation when the pregnancy, which had been complicated by a maternal rubella infection, was terminated at a regional hospital. However, vaginal bleeding persisted after the operation in spite of medication to control bleeding. Curettage of the uterine cavity one month later failed to reveal any retained placental tissue or other pathology. Therefore, an exploratory laparotomy was performed, yet nothing particular was found in the peritoneal cavity. So, the patient was transferred to our department. Sonography revealed a lower uterine mass of 4.0 x 3.3 cm in size. A persistently low serum hCG titer was also found. Placenta accreta was highly suspected. Three doses of methotrexate were given to control bleeding, yet without results. Hysterectomy was finally performed. A histological study revealed placenta increta.

Treatment and outcome of unexplained recurrent spontaneous abortions in subclinical autoimmune disorders.

Patients with a history of three or more fetal losses before a gestational age of 20 weeks were examined. Only patients with normal chromosomal, hormonal and anatomic findings were included in this study. These patients were tested for the antinuclear antibody, the C3 and C4 levels, anti-ENA [Ro(SSA), La(SSB), SM, RNP, scl-70], anti-single-stranded DNA and anti-double-stranded DNA, lupus anticoagulant, anticardiolipin and antiphosphotidylserine. All of these patients were free of any symptoms, except for the repeated abortions. The results showed that 10 out of 213 (4.7%) patients with unexplained recurrent spontaneous abortions had abnormal serologic tests, and 28% of the abortions (nine out of 32 abortions) occurred during the second trimester. With treatment of low-dose aspirin alone, or in combination with prednisolone, two out of 11 pregnancies in these 10 patients resulted in repeated abortions (18%), which was significantly lower than their previous abortion rate where 32 out of 33 pregnancies resulted in abortions (97%). Four babies (three term and one premature) were delivered without any abnormalities and the other five pregnancies are beyond the 28th week of gestation and are progressing smoothly. This study revealed that subclinical autoimmune disorders may play a role in recurrent spontaneous abortions and adequate treatment can improve the pregnancy outcome.

Pregnancy following oocyte donation and tubal embryo transfer in patients with premature ovarian failure: report of two cases.

Two pregnancies in patients with premature ovarian failure utilizing donated oocytes, in-vitro fertilization (IVF) and tubal embryo transfer (TET), are reported. The recipients received cyclic hormone replacement therapy for six months to prepare the endometrium for implantation. An evaluation cycle was tested to document that the hormone milieu established by the hormone replacement protocol was similar to that of a natural ovulatory cycle. During the oocyte donation cycle, the recipient received incremental estrogen replacement treatment of flexible length during the follicular phase of the donor's stimulated cycle to synchronize the recipient's endometrium to the donor's embryo. Concurrently, the donor underwent controlled ovarian hyperstimulation and transvaginal ultrasound-guided oocyte retrieval. After fertilization of the donated oocytes with sperm from the recipient's husband and cleavage of the fertilized oocytes into the 2- to 4-cell stage, laparoscopic embryo transfer into the recipient's fallopian tube was performed. Case 1 received 4 embryos by the TET procedure. Pregnancy was confirmed by visualization of a gestational sac in the uterine cavity 3 weeks after TET, but miscarriage occurred at the tenth gestational week. In Case 2, the pregnancy was established after TET of 2 embryos. Estrogen and progesterone supplements were maintained until day 100 after TET. The patient delivered a healthy male baby, weighing 2,520 g at 38 weeks of gestation.

The use of a long-acting gonadotropin-releasing hormone analog(D-Trp-6-LHRH) for improvement of ovarian stimulation in assisted conception programs.

To assess the efficacy of gonadotropin-releasing hormone analog (GnRHa) as an adjuvant in controlled ovarian stimulation in assisted conception programs, 114 infertile patients, who were treated by in vitro fertilization and embryo transfer (n = 61) or tubal embryo transfer (n = 53), were randomized sequentially to receive ovarian stimulation according to two protocols. In protocol 1 (n = 57), long-acting GnRHa (D-Trp-6-LHRH) microcapsules were administered intramuscularly at menstruation and ovarian stimulation using follicle-stimulating hormone (FSH) and human menopausal gonadotropin (hMG) was started 2 to 3 weeks later when the pituitary was completely suppressed. In protocol 2 (n = 57), patients received FSH and hMG from day 3 of the cycle without GnRHa pre-treatment. We found that premature luteinization did not occur in patients treated with protocol 1, and the number of cycles cancelled was also decreased. The days of ovarian stimulation and the amount of hMG required to achieve adequate follicular development were significantly higher in protocol 1 than that in protocol 2. Similarly, the mean serum estradiol levels on the day of human chorionic gonadotropin administration, number of large follicles (mean diameter greater than 10 mm), number of oocytes recovered and number of embryos obtained were also significantly higher in patients treated with protocol 1. The data suggest that the use of D-Trp-6-LHRH as an adjuvant in ovarian stimulation is associated with a lower incidence of cycle cancellation and an improvement in ovarian response in assisted conception programs.

Analysis of human in vitro fertilization failure.

Fertilization failure is a serious problem in human in vitro fertilization (IVF) programs and deserves further investigation and management. Two hundred and ninety-four IVF cycles performed at the National Taiwan University Hospital from July 1989 to June 1991 were retrospectively analyzed. Thirty-seven (13%) of the 294 cycles were observed to have fertilization failure. The incidence of fertilization failure in male factor patients was significantly higher (p < 0.05) than in others. Patients with oligoasthenospermia tended to have a higher rate of fertilization failure than patients with oligospermia or asthenospermia alone. In non-male factor patients, a smaller number of oocytes and mature oocytes was found in patients with fertilization failure than in patients achieving fertilization. Sperm penetration assay (SPA) using zona-free hamster eggs was performed for 15 patients with fertilization failure; no correlation was found between SPA and the fertilizing ability of the sperm in vitro. Four patients with severe oligoasthenospermia had repeated fertilization failure in subsequent IVF cycles. The other five patients, including four non-male factor patients and one male factor patient, achieved fertilization in their second IVF trial. Our results suggest that severe oligoasthenospermic patients with repeated fertilization failure should be candidates for micromanipulation of gametes in subsequent IVF trials.