Longitudinal assessment of anti-Müllerian hormone after cesarean section and influence of bilateral salpingectomy on ovarian reserve

ObjectiveThis study aimed to compare longitudinal changes in ovarian reserve markers after cesarean section (CS) with and without bilateral salpingectomy (BS).Study designWe prospectively enrolled women >35 weeks’ gestation scheduled for CS alone or CS + BS and obtained blood samples for anti-Müllerian hormone prior to surgery and at 3 and 6 months after surgery. At the 3-month visit, we similarly performed transvaginal ultrasound for antral follicle count.ResultsWe enrolled 50 women; 30 underwent CS only and 20 underwent CS + BS. Although anti-Müllerian hormone level increased over 6 months of follow-up in both groups, no clinically important differences in the geometric mean (interquartile range) (ng/mL) were observed at any timepoint (baseline [0.69 {0.36?1.21} {CS only} vs 0.49 {0.32?2.10} {CS + BS}, p = 0.64]; 3 months [1.35 {0.58?3.13} vs 1.45 {1.04?2.25}, p = 0.79]; and 6 months [1.74 {0.93?4.45} vs 2.60 {1.41?5.10}, p =0.27]). Similarly, we detected no difference in antral follicle count.ConclusionBS at the time of CS does not have a negative impact on ovarian reserve 6 months after surgery.ImplicationWhile our results provide reassuring data that bilateral salpingectomy for permanent contraception at the time of cesarean section does not impact ovarian reserve, longer adequately powered studies are needed.     

Activation of trans-l,2-dihydro-l,2-dihydroxy-6-aminochrysene to genotoxic metabolites by rat and human cytochromes P450

In order to address the hypothesis that 6-aminochrysene (6-AC)is converted to genotoxic products by cytochrome P450 enzymesvia two activation pathways (N-hydroxylation and epoxidation),the activation of 6-AC and trans-l,2-dihydro-l,2-dihydroxy-6-aminochrysene(6-AC-diol) to genotoxic metabolites was examined in rat andhuman liver microsomal cytochrome P450 enzymes using Salmonellatyphimurium TA1535/pSK1002 and TA1535/pSK1002/pNM12 (NM2009)as tester strains. The latter bacteria, an O-acetyl-transferase-overexpressingstrain, was highly sensitive to metabolites derived from activationof 6-AC, but not those from 6-AC-diol, using liver microsomesfrom phenobarbital-treated rats or a reconstituted monooxygenasesystem containing P4502B1 or -2B2, thus suggesting the rolesof P450 and acetyltransferase systems in the activation process.6-AC-diol, on the other hand, was activated very efficientlyby liver microsomes prepared from ß-naphthoflavone-treatedrats or a reconstituted system containing P4501A1 or -1A2; theactivation reaction is considered to proceed through diol-epoxideformation. The contribution of rat P4501A enzymes towards activationof 6-AC-diol was confirmed by the inhibitory effects on theactivation process of -naphthoflavone, a specific inhibitorof P4501 A-related activities, and antibodies raised againstpurified P4501A1 and -1A2. In humans, P4501A2 was found to bethe major enzyme involved in the activation of 6-AC-diol togenotoxic metabolites while the parent compound 6-AC was activatedmainly by P4503A4. Experiments using recombinant P450 proteinsexpressed in human lymphoblastoid cells lines showed that humanP4501A1 could also activate 6-AC-diol to reactive metabolitesat almost the same rate measured with P4501A2. In addition,P4502B6 was found to efficiently catalyze the activation of6-AC to genotoxic metabolites, and P4503A4 was active in theactivation of 6-AC-diol as well as 6-AC. Addition of purifiedrat epoxide hydrolase to the incubation mixture containing purifiedrat P4501A1 or microsomes expressing human P4501A1 caused inhibitionof activation of 6-AC-diol. These results suggest the existenceof different enzymatic activation pathways for 6-AC and 6-AC-diol.The former carcinogen may be N-hydroxylated principally by P4502Benzymes in rats and P4503A4 and -2B6 in humans and activationto its ultimate metabolites may proceed through esterificationof the N-hydroxy metabolites by an N-acetyltransferase. The6-AC-diol is metabolized to its ultimate diolepoxide productby P4501A enzymes in rat and human liver microsomes. P4503A4(humans) and P4503A2 (rats) may also contribute to some extentin the activation of 6-AC-diol, albeit at lower rates than thoseof P4501A enzymes.     

Establishment of 2 human thyroid-carcinoma cell-lines (8305c, 8505c) bearing p53 gene-mutations

New cell lines, designated 8305C and 8505C, were established from undifferentiated thyroid carcinomas of a 67 year-old-female patient and a 78-year-old-female patient, respectively. Pathologically both these primary undifferentiated carcinoma tissues contained residual well differentiated components, suggesting well differentiated to undifferentiated carcinoma progression. Cell kinetic analysis indicate that the cell population doubling time is 43 h for 8305C and 36 h for 8505C. The saturation density at confluency is 5.7 x 10(4) cells/cm2 for 8305C and 1.1 x 10(5) cells/cm2 for 8505C. To identify genetic changes that may have occurred in these two cell lines, tumor suppressor genes p53, Rb, APC and MCC were analyzed. Sequence analysis confirmed a C:G to T:A transition at the first base of p53 gene codon 273 in 8305C and a C:G to G:C transversion at the first base of p53 codon 248 in 8505C. Polymerase chain reaction-loss of heterozygosity assays confirmed allelic deletion of p53 gene from the 8505C cell line. Loss of heterozygosity of other tumor suppressor genes were not observed. Given that p53 mutations associate with undifferentiated carcinoma but not with well differentiated carcinoma during multistep carcinogenesis of the thyroid, these cell lines should prove useful for research into the role of p53 gene mutations in malignant transformation.     

Human erythrocyte band 3 (EPB3) polymorphism: Analysis of blood and bloodstains by an immunodetection method and frequency of the EPB3* Memphis variant

Summary The erythrocyte band 3 (EPB3) variant, band 3 Memphis (EPB3*Memphis), was detected by immunoblotting with a monoclonal antibody to the 41 kDa cytoplasmic N-terminal domain of band 3 without protease treatment of erythrocytes. EPB3*Memphis was also detected by immunoblotting from 3-month-old bloodstains subjected to -chymotrypsin treatment. A population genetic study using this method indicated that the EPB3 variant would be useful for forensic work in Japan, since the frequency of this variant in Japanese (Wakayama prefecture) is relatively high (0.159).     

Refilling the lens with an inflatable endocapsular balloon: surgical procedure in animal eyes

In this report we describe the surgical details involved in refilling the lenses of 13 rabbit and 3 primate eyes using an inflatable endocapsular balloon to restore accommodation. The procedure involves endocapsular phacoemulsification through a small buttonhole or dumbbell anterior capsulotomy or minicircular capsulotomy and the simultaneous preservation of capsular integrity, including the zonules and ciliary muscles. An inflatable balloon made of thin silicone membrane is then inserted into the empty capsular bag. A liquid silicone polymer is injected into the balloon through a delivery tube, and the empty capsular bag is refilled by the inflated balloon. The procedure was found to be reproducible, and an accommodation of 6 D was confirmed in one primate eye. Capsular opacification occurred, but the proliferation and migration of residual lens epithelial cells could be hindered by abundant refilling. This lens-refilling technique may provide restoration of accommodation in future cataract surgery. Offprint requests to: O. Nishi     

Does Hyperthermia Induce Peritoneal Damage in Continuous Hyperthermic Peritoneal Perfusion?

To investigate the mechanisms of the peritoneal damage induced by continuous hyperthermic peritoneal perfusion (CHPP), protein and fluid loss during and after CHPP and continuous normothermic peritoneal perfusion (CNPP) was studied. Sixteen patients with advanced gastric cancer underwent peritoneal perfusion therapy with saline solution containing 150 to 300 mg cisplatin and 30 to 60 mg mitomycin C for 60 minutes. The temperature in Douglas' pouch was maintained at 42.0°C in the CHPP group (n= 9) and 37.0°C in the CNPP group (n= 7) during perfusion. No statistical differences were found in patients' characteristics between the groups except the maximum temperature in Douglas' pouch during perfusion (41.6°± 0.4°C and 37.6°± 0.4°C in CHPP and CNPP groups, respectively, p < 0.05). The amount of protein lost into the perfusate was 0.35 ± 0.22 g/kg body weight in the CHPP group and 0.37 ± 0.19 g/kg in the CNPP group, showing no significant difference. On the day of surgery, there was no significant difference in the amount of protein and fluid lost through the abdominal drains between the CHPP group (27.9 ± 24.6 mg/kg/hr and 0.94 ± 0.63 ml/kg/hr, respectively) and the CNPP group (25.9 ± 8.6 mg/kg/hr and 1.03 ± 0.31 ml/kg/hr, respectively). We could not find any significant differences in postoperative protein and fluid loss between the groups on the following 3 days either. We conclude that the peritoneal damage by CHPP is not caused by the hyperthermia but by the peritoneal perfusion with saline solution containing anticancer drugs.     

Roles of human CYP2A6 and 2B6 and rat CYP2C11 and 2B1 in the 10-hydroxylation of (-)-verbenone by liver microsomes.

(-)-Verbenone, a monoterpene bicyclic ketone, is a component of the essential oil from rosemary species such as Rosmarinus officinalis L., Verbena triphylla, and Eucalyptus globulus and is used for an herb tea, a spice, and a perfume. In this study, (-)-verbenone was found to be converted to 10-hydroxyverbenone by rat and human liver microsomal cytochrome p450 (p450) enzymes. The product formation was determined by high-performance liquid chromatography with UV detection at 251 nm. There was a good correlation between activities of coumarin 7-hydroxylation and (-)-verbenone 10-hydroxylation catalyzed by liver microsomes of 16 human samples, indicating that CYP2A6 is a principal enzyme in (-)-verbenone 10-hydroxylation in humans. Human recombinant CYP2A6 and CYP2B6 catalyzed (-)verbenone 10-hydroxylation at Vmax values of 15 and 21 nmol/min/nmol p450 with apparent Km values of 16 and 91 microM, respectively. In contrast, rat CYP2A1 and 2A2 did not catalyze (-)-verbenone 10-hydroxylation at all, suggesting that there were species-related differences in the catalytic properties of human and rat CYP2A enzymes in the metabolism of (-)-verbenone. In the rat, recombinant CYP2C11, CYP2B1, and CYP3A2 catalyzed (-)-verbenone 10-hydroxylation with Vmax and Km ratios (ml/min/nmol p450) of 0.73, 0.20, and 0.03, respectively. Male-specific CYP2C11 was a major enzyme in (-)-verbenone 10-hydroxylation by untreated rat livers, and CYP2B1 catalyzed this reaction in liver microsomes of phenobarbital-treated rats. Rat CYP2C12, a female-specific enzyme, did not catalyze (-)verbenone 10-hydroxylation. These results suggest that human CYP2A6 and rat CYP2C11 are the major catalysts in the metabolism of (-)-verbenone by liver microsomes and that there are species-related differences in human and rat CYP2A enzymes and sex-related differences in male and female rats in the metabolism of (-)-verbenone.     

The influence of androgen deprivation therapy on dihydrotestosterone levels in the prostatic tissue of patients with prostate cancer.

PURPOSE: The influence of androgen deprivation therapy on dihydrotestosterone levels in the prostatic tissue is not clearly known. Changes in dihydrotestosterone levels in the prostatic tissue during androgen deprivation therapy in the same patients have not been reported. We analyzed dihydrotestosterone levels in prostatic tissue before and after androgen deprivation therapy. EXPERIMENTAL DESIGN: A total of 103 patients who were suspected of having prostate cancer underwent prostatic biopsy. Sixty-nine patients were diagnosed as having prostate cancer whereas the remaining 34 were negative. Serum samples were collected before biopsy or prostatectomy. Dihydrotestosterone levels in prostatic tissue and serum were analyzed using liquid chromatography/electrospray ionization-mass spectrometry after polar derivatization. In 30 of the patients with prostate cancer, dihydrotestosterone levels in prostatic tissue were determined by performing rebiopsy or with prostate tissues excised after 6 months on androgen deprivation therapy with castration and flutamide. RESULTS: Dihydrotestosterone levels in prostate tissue after androgen deprivation therapy remained at approximately 25% of the amount measured before androgen deprivation therapy. Dihydrotestosterone levels in serum decreased to approximately 7.5% after androgen deprivation therapy. The level of dihydrotestosterone in prostatic tissue before androgen deprivation therapy was not correlated with the serum level of testosterone. Serum levels of adrenal androgens were reduced to approximately 60% after androgen deprivation therapy. CONCLUSIONS: The source of dihydrotestosterone in prostatic tissue after androgen deprivation therapy involves intracrine production within the prostate, converting adrenal androgens to dihydrotestosterone. Dihydrotestosterone still remaining in prostate tissue after androgen deprivation therapy may require new therapies such as treatment with a combination of 5alpha-reductase inhibitors and antiandrogens, as well as castration.     

Somatic Mutations of the PTEN/MMAC1 Gene in Fifteen Japanese Endometrial Cancers: Evidence for Inactivation of Both Alleles

Loss of heterozygosity (LOH) of chromosome 10q is observed in approximately 40% of endometrial cancers. Mutations in PTEN/MMAC1 , a gene recently isolated from the 10q23 region, are responsible for two dominantly inherited neoplastic syndromes, Cowden disease and Bannayan-Zonana syndrome. Somatic mutations of this gene have also been detected in sporadic cancers of the brain, prostate and breast. To investigate the potential role of this putative tumor suppressor gene in endometrial carcinogenesis as well, we examined 46 primary endometrial cancers for LOH at the 10q23 region, and for mutations in the entire coding region and exon-intron boundaries of the PTEN/MMAC1 gene. LOH was identified in half of the 38 informative cases, and subtle somatic mutations were detected in 15 tumors (33%). Our results suggest that of the genes studied so far in endometrial carcinomas, PTEN/MMAC1 is the most commonly mutated one, and that inactivation of both copies by allelic loss and/or mutation, a pattern that defines genes as "tumor suppressors,'contributes to tumorigenesis in endometrial cancers.