Inhibitory effect of 5ß-pregnane-3α,20ß-diol on transcriptional activity and enzyme activity of human bilirubin UDP-glucuronosyltransferase.

Prolonged unconjugated hyperbilirubinemia in infants associated with breast milk feeding is a common pediatric problem known as breast milk jaundice (BMJ). A polymorphic mutation (G71R) of bilirubin UDP-glucuronosyltransferase (UGT1A1) is a known cause of BMJ on the infantile side, but the responsible components of breast milk are not currently known. We analyzed the inhibitory effect of 5ß-pregnane-3α,20ß-diol (pregnanediol) on transcriptional activity and enzyme activity of UGT1A1. To this end, we constructed two types of expression vectors. One type comprised vectors including the upstream enhancer-promoter sequence of UGT1A1 for WT and variant type (-3279T>G with A(TA)7TAA), used in studying transcriptional regulation. The other type comprised vectors including cDNA of UGT1A1 for WT and the G71R variant, used in studying enzyme activity. In an in vitro expression study, pregnanediol did not affect the transcriptional activity of UGT1A1 enhancer-promoter complex of WT and variant type, even with coexistence of transcriptional factors such as constitutive androstane receptor or pregnane X receptor. In contrast, in the presence of 100 µM pregnanediol, bilirubin glucuronidation of G71R-UGT1A1 was reduced to 51% of WT. We suggest that pregnanediol is a cause of breast milk jaundice in carriers of G71R.

Transient congenital hypothyroidism caused by biallelic mutations of the dual oxidase 2 gene in Japanese patients detected by a neonatal screening program.

CONTEXT: Mutations in dual oxidase (DUOX2) have been proposed as a cause of congenital hypothyroidism. Previous reports suggest that biallelic mutations of DUOX2 cause permanent congenital hypothyroidism and that monoallelic mutations cause transient congenital hypothyroidism. OBJECTIVE: To clarify the inheritance of hypothyroidism, we looked at the DUOX2 gene in patients with transient congenital hypothyroidism. DESIGN: DUOX2, thyroid peroxidase, Na+/I- symporter and dual oxidase maturation factor 2 genes were analyzed in eight patients with transient congenital hypothyroidism, using the PCR-amplified direct sequencing method. PATIENTS: The eight patients were found by a neonatal screening program. Six of these patients belonged to two independent families; the other two were unrelated. Their serum TSH values varied from 24.8-233.0 mU/liter. Six of the eight patients had a low serum freeT4 level (0.19-0.84 ng/dl). Seven of the eight patients were treated with thyroid hormone replacement therapy, which ceased to be necessary by 9 yr of age. RESULTS: Eight novel mutations were detected in the DUOX2 gene. Four patients in one family were compound heterozygous for p.L479SfsX2 and p.K628RfsX10. Two patients in a second family were compound heterozygous for p.K530X and p.[E876K;L1067S]. The two remaining unrelated patients were also compound heterozygous, for p.H678R/p.L1067S and p.A649E/p.R885Q, respectively. CONCLUSION: All eight patients had biallelic mutations in the DUOX2 gene. We find that loss of DUOX2 activity results in transient congenital hypothyroidism and that transient congenital hypothyroidism caused by DUOX2 mutations is inherited as an autosomal recessive trait.

Changes in urinary excretion of six biochemical parameters in normotensive pregnancy and preeclampsia.

We evaluated renal functions by urinary biochemical parameters in normotensive pregnancy and preeclampsia. The parameters are expected to be altered resulting from different abnormalities of renal glomeruli and tubules. We chose N-acetyl-beta-d-glucosaminidase (NAG), beta2-microglobulin (beta2MG), total protein (TP), albumin (Alb), urea nitrogen (UN), uric acid (UA), and creatinine (Cr). Urinary excretion of these biochemical parameter concentrations (relative to Cr) was measured simultaneously in first morning fasting urine samples from 27 healthy nonpregnant women (group 1), 32 women with normotensive pregnancies (group 2), and 26 women with preeclampsia (group 3). The average gestational age at entry was 36 weeks. Serum UN and serum UA also were measured. All the ratios were significantly higher in group 2 than in group 1. The NAG-to-Cr, TP-to-Cr, and Alb-to-Cr ratios were significantly higher in group 3 than in group 2. In contrast, the UN-to-Cr and UA-to-Cr ratios were significantly lower in group 3 than in group 2. The percent increase in the beta2MG-to-Cr ratio in group 2 relative to that in group 1 was the highest, followed by percent increases in the NAG-to-Cr, TP-to-Cr, Alb-to-Cr, UA-to-Cr, and UN-to-Cr ratios. In contrast, the percent increase in the Alb-to-Cr ratio in group 3 relative to that in group 2 was the highest, followed by percent increases in the TP-to-Cr, NAG-to-Cr, beta2MG-to-Cr, UA-to-Cr, and UN-to-Cr ratios. The percent increases in the NAG-to-Cr and beta2MG-to-Cr ratios rose markedly in normotensive pregnancy, whereas percent increases of the Alb-to-Cr and TP-to-Cr ratios were far greater in preeclampsia than in normotensive pregnancy. Renal tubular damage and reabsorption dysfunction may be impaired markedly even in normotensive pregnancy, and further deterioration in reabsorption dysfunction may be slight in preeclampsia. Renal glomerular permeability of TP and Alb may be enhanced in normotensive pregnancy and markedly enhanced in preeclampsia.

Plasma levels of alpha-defensins 1-3 are an indicator of neutrophil activation in pregnant and post-partum women.

AIM: In severe preeclampsia and septic shock, excessively activated neutrophils are thought to injure tissue irreversibly. On the other hand, mild neutrophil activation is known to occur during normal pregnancy. The objective of this study was to determine whether elevated plasma levels of alpha-defensins 1-3 could be used as an indicator of neutrophil activation in pregnant and post-partum women. METHODS: Defensin concentrations in 21 non-pregnant women and men, 184 normal pregnant women, and 55 post-partum women were quantified using an enzyme-linked immunosorbent assay (ELISA). The expression of the surface markers, CD11b and Toll-like receptor-4 (TLR-4), on the neutrophils were analyzed by flow cytometry in a cohort of subjects different from that used for the analysis of alpha-defensin levels. RESULTS: The concentrations of alpha-defensins were significantly higher in women that were in labor than in any of the other subjects. These levels diminished after delivery, but remained significantly elevated at one month post-partum. The expression of both CD11b and TLR-4 was significantly higher in women in labor compared to non-pregnant donors (controls). CD11b expression remained high on the third post-partum day, while TLR-4 expression fell to non-pregnant levels. CONCLUSION: Our results suggest that there is a positive association between defensin levels and neutrophil activation in pregnant and post-partum women.

Inhibitory effects of herbal drugs on the growth of human ovarian cancer cell lines through the induction of apoptosis.

OBJECTIVE: In order to develop and search for more effective and safe treatments for early and advanced stages of ovarian cancer, we examined the direct effects of four extracts of Chinese herbal drugs on ovarian cancer cells in vitro. METHODS: The growth inhibition of four herbal drugs on a total of six cell lines of human ovarian cancer cells was determined by a Cell Counting Kit-8 by counting viable cells. Apoptotic cells induced by herbal drugs were detected by using MEBCYTO Apoptosis Kit. All experiments were performed in triplicate. The significance of the difference was analyzed with a two-sided Student's t test. A P value less than 0.05 was accepted as statistically significant. RESULTS: The MN, A2780, and KF cell lines exhibited significant growth inhibition in the presence of Sho-saiko-to concentrations of 150 microg/ml, 300 microg/ml, and 500 microg/ml, respectively, and at the concentration of 1000 microg/ml, Sho-saiko-to demonstrated a significant apoptotic induction effect on all six kinds of ovarian cancer cell lines. This concentration is the same as the blood concentration attained when 7.5 g of Sho-saiko-to per day is orally administered and all absorbed. CONCLUSIONS: Sho-saiko-to exhibited significant growth inhibition of ovarian cancer cell lines, and the mechanisms of the inhibitory effects can be attributed, in part, to apoptosis induced by Sho-saiko-to.

A case of T1N0M0 vulvar apocrine gland carcinoma with a positive outcome.

BACKGROUND: Apocrine carcinoma of the vulva is extremely rare; only two cases have been reported worldwide. Here we report a case of apocrine carcinoma of the vulva. CASE: A 58-year-old woman complaining of a small, asymptomatic genital tumor visited the gynecology clinic of Chiba Social Insurance Hospital. The biopsy specimen suggested that it was an adenocarcinoma derived from the apocrine gland. The diagnosis was confirmed by periodic acid-Schiff staining and immunohistochemical staining for gross cystic disease fluid protein 15. She underwent simple vulvectomy with hemilateral groin dissection. She has been followed as an outpatient for the past 7 years with no signs of recurrence or metastasis. CONCLUSION: Despite the positive outcome of this case, small, asymptomatic genital lesions should be regarded with caution.

The relationship between telomere length and telomerase activity in gynecologic cancers.

OBJECTIVE: In recent years, many studies have been published regarding telomere length and telomerase activity in malignant tissues. However, it is not enough that telomere length and telomerase activity in gynecologic cancers have been measured at same time. We investigated the relationship between telomere length and telomerase activity in gynecologic cancers. METHODS: A total of 52 gynecologic cancers (15 ovarian cancers, 23 endometrial cancers, 14 cervical cancers) were obtained at the time of surgery. The specimens were analyzed for telomerase activity and telomere length with the TRAP(EZE) ELISA kit and Southern blot, respectively. RESULTS: Telomerase activity was detected in 42 of 50 (84.0%) of all evaluable specimens, in 11/15 (73.3%) ovarian, 18/22 (81.8%) endometrial, and in 13/13 (100%) cervical cancers. The difference of positive strength (DeltaA value) between stage I and III was statistically significant (P = 0.01, ANOVA test). Changes in telomere length by shortening or elongation were detected in 35 of 52 (67.3%) tumors, in 9/15 (60.0%) ovarian, 17/23 (73.9%) endometrial, and 9/14 (64.3%) cervical cancers. There was no detectable relationship between telomere length and stage of disease, pathologic diagnosis (ovarian, endometrial, and cervical cancers), or telomerase activity. CONCLUSIONS: There was no relationship between telomerase activity and telomere length. The clinical significance of telomere length appears to be limited in gynecologic cancers.