Successful trachelectomy and re-vaginoplasty for cervico-vaginal stenosis following unsuccessful uterovaginal anastomosis and vaginoplasty in congenital cervical and vaginal aplasia: description of technique and a case report.

We describe a successful surgical technique of abdominal trachelectomy and re-vaginoplasty for cervico-vaginal stenosis following unsuccessful uterovaginal anastomosis and vaginoplasty in a patient with congenital cervical and vaginal aplasia. After the surgical procedure, cervico-vaginal stenosis was resolved and periodic menstruation without dysmenorrhoea resumed. While long-term follow-up is essential to ensure successful pregnancy and delivery, we conclude that this novel surgical procedure is a promising alternative for improvement of the quality of life and normal sexual function, and for preservation of fertility in patients with cervical and vaginal aplasia.

Pathological factors associated with omental metastases in endometrial cancer.

PURPOSE OF INVESTIGATION: This study aimed to assess the role of omentectomy in the surgical therapy of endometrial cancer. MATERI- ALS AND METHODS: A retrospective study was performed in 98 patients who were pathologically diagnosed with endometrial cancer and had initially undergone surgical therapy at the present institution. This study analyzed the relationship between omental metastasis and clinicopathological factors. RESULTS: Omental metastasis was detected in nine patients (9%). On univariate analysis, significant number of omental metastatic lesions were detected in few cases by positive peritoneal cytology, adnexal metastasis, gross dissemination, and lymphovascular space involvement. On multivariate analysis, adnexal metastasis were a significant risk factor. The sensitivity of the spe- cial histological type and the specificity of the macroscopic peritoneal dissemination and adnexal metastasis were all high. CONCLUSION: Omentectomy plays a significant role in determining the exact surgical staging in cases with non-endometrioid cancer, adnexal metas- tasis, and macroscopic peritoneal dissemination.

Molecular epidemiological and mutational analysis of DNA mismatch repair (MMR) genes in endometrial cancer patients with HNPCC-associated familial predisposition to cancer.

Recently, a high rate of endometrial cancer has been reported in women with hereditary non-polyposis colorectal cancer (HNPCC), suggesting a relationship between familial endometrial cancers and HNPCC. Familial endometrial cancers constitute only about 0.5% of all endometrial carcinomas and it is essential to examine family histories in detail. A mutational analysis of three DNA mismatch repair (MMR) genes (hMLH1, hMSH2 and hMSH6) in patients with endometrial cancer who meet our criteria for familial predisposition to HNPCC-associated endometrial cancers was performed. Mutations were detected in 18 of the 120 patients (15.0%). Most HNPCC-related endometrial cancers do not meet the New Amsterdam Criteria for HNPCC. These clinical criteria may identify only some HNPCC-associated endometrial cancers. Establishing the correct family history for endometrial cancer patients is important for diagnosing familial endometrial carcinomas. An analysis of MMR genes may be useful for patients with endometrial cancer showing familial aggregation. In addition, gynecologists must be accurately informed, and it is important to perform large-scale, multicenter studies both nationwide and internationally.

Can ABCF2 protein expression predict the prognosis of uterine cancer?

Uterine cervical and endometrial cancers are common malignant solid neoplasms for which there are no useful prognostic markers. In this study, we evaluate the relationship between ATP-binding cassette superfamily F2 (ABCF2) expression and clinical factors including clinical stage, histologic type, grade and prognosis in uterine cervical and endometrial cancer. Two hundred and sixty seven cervical and 103 endometrial cancers were studied. ATP-binding cassette superfamily F2 cytoplasmic expression was detected by immunohistochemical staining and scored as positive or negative. Among cervical cancer cases, 149 (55.8%) expressed ABCF2. The overall survival was longer in ABCF2-negative than ABCF2-positive cases (P=0.0069). Statistically significant prognostic factors for survival were ABCF2 positivity (risk ratio (rr)=1.437), old age (rr=1.550) and advanced stage (rr=2.577). ATP-binding cassette superfamily F2 positivity was an independent prognostic factor by multivariate proportional hazard test (P=0.0002). Among endometrial cancer cases, 72 (69.9%) were cytoplasmic ABCF2 positive. However, there was no significant relationship between ABCF2 expression and age, clinical stage, histologic type, histologic grade, oestrogen receptor status or prognosis. ATP-binding cassette superfamily F2 expression may be a useful prognostic marker in cervical but not endometrial cancer. The role of ABCF2 protein may differ depending on the type of cancer.

Current Status of Uterus Transplantation and Approaches for Future Clinical Application in Japan.

Uterus transplantation (UTx) has become an alternative to gestational surrogacy and adoption for women with uterine factor infertility (UFI). Brännström et al achieved the first human delivery after UTx in 2014, and to date a total of 8 babies have been born after UTx from living donors. This outcome has attracted much attention worldwide, and many countries are now preparing for UTx. There are an estimated 60,000 women of reproductive age with UFI in Japan, and these patients cannot have biological children because gestational surrogacy is forbidden in Japan. We have performed UTx research from 2009 using cynomolgus macaque, in preparation for clinical application of UTx for these patients to have a child, and we have accumulated a large amount of data. However, the UTx procedure still has many medical, ethical, and social issues that require discussion prior to clinical application. The Japan Society for Uterus Transplantation was established in 2014 for further discussion of these issues in Japan. UTx is still in the experimental stage overseas, and the safety and efficacy remain unclear, despite several clinical applications. Despite the many issues to be resolved, this organ transplantation technology will provide new hope for women with UFI, and further development of the technology is important for future reproductive and transplant medicine. In this article, we summarize the current status of UTx and the situation regarding future clinical application in Japan.

Organ Perfusion for Uterus Transplantation in Non-Human Primates With Assumed Procurement of a Uterus From a Brain-Dead Donor.

BACKGROUND: Clinical studies of uterus transplantation have been performed to treat uterine factor infertility. Because the uterus is a pelvic visceral organ, the method of perfusion for the procurement of vital organs from a brain-dead donor should be modified for removal of the uterus. Herein, we report the results of a preliminary study in cynomolgus monkeys of a new perfusion method for uterus transplantation with assumed procurement of a uterus from a brain-dead donor. METHODS: Cynomolgus monkeys were used; thoracolaparotomy was performed on the donor. A perfusion catheter was then placed into the unilateral femoral artery and/or external iliac artery. Cross-clamping was performed for the aorta under the diaphragm and the inferior vena cava was divided in the pleural space. The perfusion solution was then administered via the catheter to perfuse all organs in the abdominal cavity, including those in the pelvic cavity. After the perfusion, gross observation and histopathological examination of abdominal organs were conducted. RESULTS: Gross findings showed that all abdominal organs turned white in all specimens, indicating favorable perfusion of the uterus and all other organs in the abdomen. Pathological findings showed that almost no hemocytes were observed in the vessels of each organ. CONCLUSIONS: With perfusion via the femoral artery and/or external iliac artery, all organs in the abdominal cavity, including the uterus, could be perfused. It was suggested that this technique could be useful for uterus transplantation assuming the procurement of a uterus from a brain-dead donor.

Pharmacological properties of N-(3',4'-dimethoxycinnamoyl) anthranilic acid (N-5'), a new anti-atopic agent.

1 N-(3'-4'-dimethoxycinnamoyl) anthranilic acid (N-5') exhibited a dose-dependent, potent inhibition of the passive cutaneous anaphylaxis (PCA) mediated by homocytotropic antibodies (HTA), which was hardly affected by anti-inflammatory agents such as phenylbutazone, indomethacin and prednisolone at any dose used. The HTA-induced PCA was significantly inhibited by combined treatment with diphenydramine and cyproheptadine. 2 Doses of N-5' which potently inhibited HTA-induced PCA inhibited only slightly the heterologous PCA produced by anti-bovine serum albumin (BSA) rabbit serum. This heterologous PCA was clearly inhibited by phenylbutazone, indomethacin and prednisolone. Diphenydramine and cyproheptadine, singly or combined inhibited the heterologous PCA only slightly. 3 The increased vascular permeability caused by histamine and 5-hydroxytryptamine was significantly inhibited by diphenyldramine or cyproheptadine, but not by N-5' and the anti-inflammatory agents used. 4 N-5' 150 mg/kg orally inhibited rat paw oedema induced by carrageenin by about 26% while phenylbutazone, indomethacin and prednisolone produced significant inhibition. 5 N-5' at concentrations of 100 and 1000 muM significantly inhibited (by about 52% and 95%, respectively) the histamine release from rat peritoneal cells induced by HTA; 10 muM N-5' had little effect. Histamine release was inhibited by phenylbutazone or indomethacin at 1000 muM but not at 100 muM. Prednisolone had no effect on histamine release at any of the concentrations used. 6 These findings suggest that the inhibition of the HTA-induced PCA by N-5' may be due to inhibition of histamine release and is clearly different from the actions of anti-inflammatory agents such as phenylbutazone, indomethacin and prednisolone.

Effects of corticotropin-releasing factor on feeding and pancreatic polypeptide response in the dog.

We have previously reported that corticotropin-releasing factor (CRF) is a potent stimulator of adrenocorticotropic hormone and cortisol secretion in the dog. Therefore, in the present study, we investigated the extrahypophysiotropic actions of CRF in this species. When CRF was injected into the third cerebral ventricle, it failed to inhibit food intake significantly at doses of 1.19, 3.57, and 11.9 nmol. This is in sharp contrast with the results in rodents. At the 3.57 and 11.9 nmol doses, CRF markedly stimulated the secretion of pancreatic polypeptide (PP), a hormone under vagal control, and at the highest dose CRF increased plasma glucose levels. These results suggest species differences in the feeding response to CRF and activation of the parasympathetic nervous system in the dog.

Major glycoproteins in carp CNS myelin: homology to P0 protein with HNK-1/L2 carbohydrate epitope.

We studied the myelin protein profiles of carp from a phylogenetic point of view. The carp central nerve myelin contained two reactive bands, 28 and 25 kDa, demonstrated with anti-bovine P0 antibody. Their molecular weights are slightly different from those of two positive bands found in carp peripheral myelin. The N-terminal amino acid sequences of these four positive bands were identical to one another and showed high homology with those of mammalian P0 protein, suggesting that carp central myelin contains the P0-like protein. Lectin binding analysis revealed that carbohydrate structure of the P0-like proteins in carp central myelin is similar to those in peripheral myelin of carp and other vertebrates. Further, the carp P0-like glycoproteins, like the P0 proteins of other vertebrates, reacted with antibodies that recognize the HNK-1/L2 carbohydrate epitope. We conclude that the major structural glycoprotein in central myelin of the carp is homologous to P0 protein in peripheral myelin of other higher classes of vertebrates.

Prognosis after hepatic resection in patients with hepatocellular carcinoma, estimated on the basis of the morphometric indices.

To determine whether the morphometric indices of hepatocellular carcinoma (HCC) correlated with the prognoses, the microscopic morphometric values for 84 HCC cases treated by hepatic resection were studied using an image analyzer in relation to the survival rate and the gross classification. The mean survival time (MST) was 58 months in cases with a nucleocytoplasmic area ratio (N/C) of less than 0.28; this was significantly longer than the 38-month MST in cases with an N/C of more than 0.28 (P < 0.05). In stage III disease, the MST for cases with an N/C of less than 0.28 was 63 months, which was significantly longer than the MST of 13 months for cases with an N/C of more than 0.28. After relatively noncurative hepatic resection, the MST for cases with an N/C of less than 0.28 was 49 months, and this was significantly longer than the MST of 8 months for cases with an N/C of more than 0.28. The MST was 71 months for cases with a coefficient of variance of the nuclear form factor (NCV) of less than 5.5%, which was significantly longer than the MST of 33 months for cases with an NCV of more than 5.5% (P < 0.05). In stage III disease, the MST was 69 months for cases with an NCV of less than 5.5%, and this was significantly longer than the MST of 29 months for cases with an NCV of more than 5.5% (P < 0.05). In cases with an N/C of less than 0.28, 18% had vascular invasion and 38% had intrahepatic metastases, whereas in those with an N/C of more than 0.28, 62% had vascular invasion and 67% had intrahepatic metastases (P < 0.01, P < 0.05). Based on the results of these morphometric studies on HCC cases treated by hepatic resection, N/C and NCV may be useful as prognostic factors.

Steady-state pharmacokinetics and pharmacodynamics of imidaprilat, an active metabolite of imidapril, a new angiotensin-converting enzyme inhibitor in spontaneously hypertensive rats.

Imidapril, a new angiotensin-converting enzyme (ACE) inhibitor, was infused subcutaneously at the rates of 9, 30, 90, and 300 micrograms/rat/day for 4 weeks via an osmotic pump implanted under the skin in the back of male spontaneously hypertensive rats (SHRs). Plasma concentrations of imidaprilat as an active metabolite of imidapril, systolic blood pressure (SBP), and plasma ACE activity were determined periodically. These results were also compared with those of enalapril. The plasma concentrations of an active metabolite of both the imidapril and enalapril groups increased according to the doses and showed almost the same plasma concentrations at the same doses. Both groups significantly inhibited plasma ACE activity and reduced SBP, and these actions were maintained for 4 weeks. At the lowest dose studied (9 micrograms/rat/day), imidapril was more potent than enalapril in inhibiting plasma ACE (maximum 2.5-fold difference), but this difference was reduced at higher doses. In contrast, significant differences in SBP effects were observed only at the highest dose studied (300 micrograms/rat/day). Also, the imidapril group significantly decreased the relative heart weight at the rate of 300 micrograms/rat/day. Furthermore, good correlations between plasma imidaprilat concentration and plasma ACE activity or SBP were observed, suggesting that plasma concentration may be a useful marker of pharmacological effects. However, a poor relationship between plasma ACE activity and SBP for enalapril was observed, suggesting that this may not be an adequate marker of pharmacologic efficacy of ACE inhibitors in general. The clinical relevance of these findings is not known at present.

Pharmacokinetic and pharmacodynamic study of imidaprilat, an active metabolite of imidapril, a new angiotensin-converting enzyme inhibitor, in spontaneously hypertensive rats.

The pharmacokinetics and pharmacodynamics (PK/PD) of imidaprilat, an active metabolite of imidapril, a new angiotensin-converting enzyme (ACE) inhibitor, were investigated. Imidapril was infused subcutaneously for 4 weeks via an osmotic pump implanted under the skin in the back of male spontaneously hypertensive rats (SHRs). Plasma concentration of imidaprilat, systolic blood pressure (SBP), and plasma ACE activity were determined periodically. The plasma concentration of imidaprilat increased in proportion to the infusion rates and was maintained for 4 weeks. The SBP and ACE activity did not decrease in proportion to the infusion rates due to the saturation of the pharmacologic effects, but these actions also were maintained for 4 weeks. The PK/PD of imidaprilat were not influenced by aging of SHRs. The antihypertensive action in subcutaneous infusion of imidapril was as potent as that in oral administration at the same dose, although the maximum plasma concentration of imidaprilat in subcutaneous infusion was one-eightieth times of that in oral administration. The action was also maintained 28 times longer than that in oral administration, indicating that subcutaneous infusion is useful as an administration route. Furthermore, good correlation between plasma imidaprilat concentration and SBP was observed in subcutaneous infusion, indicating that plasma concentration may be a useful marker of pharmacologic action.

Pharmacokinetics and pharmacodynamics of a sustained-release biodegradable pellet containing imidapril, a new angiotensin-converting enzyme inhibitor in spontaneously hypertensive rats.

The pharmacokinetics and pharmacodynamics (PK/PD) of a sustained-release biodegradable pellet containing imidapril, a new angiotensin-converting enzyme (ACE) inhibitor, were investigated in comparison with those of an osmotic pump in male spontaneously hypertensive rats (SHRs). A pellet was prepared from copolymer of DL-lactic acid and glycolic acid by the melt-pressing technique. Imidapril was released in vitro from the pellet at an approximately zero-order rate and the release profile was similar to that of the osmotic pump. Imidapril was administered subcutaneously via a pellet or an osmotic pump implanted under the skin in the back of SHRs. Plasma concentrations of imidaprilat as an active metabolite of imidapril, plasma ACE activity and systolic blood pressure (SBP) were determined periodically. The plasma concentration of imidaprilat during the administration of a pellet was maintained for 4 weeks, and the plasma concentration profile was close to that of the osmotic pump. Both groups of pellet and osmotic pump significantly inhibited plasma ACE activity and reduced SBP for 4 weeks, and these action profiles were similar in both groups. In addition, in vivo release profile of the pellet was close to the in vitro release profile, and the in vivo release profiles of the pellet and the osmotic pump were similar to each other. From these results, it was found that the PK/PD of a biodegradable pellet were close to those of the osmotic pump, and it was shown that the pellet may be a useful system to maintain the plasma concentration of imidaprilat for a long time.

Simultaneous determination of an active metabolite and open-ring metabolites by high performance liquid chromatography and pharmacokinetic studies of a penem antibiotic, FCE22891, in dogs.

A sensitive high performance liquid chromatographic method for the simultaneous determination of an active metabolite (FCE22101) and open-ring metabolites (P1, P2) of a penem antibiotic, FCE22891, in dog plasma was developed. Plasma samples were pretreated only by ultrafiltration for the determination of the metabolites. The filtrates were directly analyzed by a reversed-phase high-performance liquid chromatographic system using a two-sided bracketing injection technique. The quantitation limits of FCE22101, P1 and P2 were 0.03, 0.1 and 0.15 microgram ml-1, respectively. Analysis of the spiked plasma samples demonstrated the good accuracy and precision of the method. The proposed method was applied to the pharmacokinetic studies of an active metabolite and open-ring metabolites after oral administration of a penem antibiotic, FCE22891, in dogs. In addition, the plasma levels of unchanged FCE22891 and the possible changes of formaldehyde and acyl-L-carnitine levels in plasma, which will be generated from the ester group of FCE22891, were also investigated.

Radioimmunoassay for TA-0910, a new stable thyrotropin releasing hormone analogue and its metabolite, TA-0910 acid-type, in human plasma and urine.

Radioimmunoassay (RIA) was investigated for the determination of TA-0910 and its main metabolite, TA-0910 acid-type, in human plasma and urine. TA-0910 is a new metabolically stable analogue of thyrotropin releasing hormone (TRH). Antiserum was raised in the rabbit against the 1-fluoro-2,4-dinitrophenyl derivative of TA-0910 or TA-0910 acid-type conjugated to keyhole limpet hemocyanin (KLH). The radioligand was prepared by iodination with 125I of the histidine imidazole ring of TA-0910 or TA-0910 acid-type. Cross-reactivities of anti-TA-0910 or TA-0910 acid-type antiserum for TA-0910, its metabolite and related compounds were low. The calibration range was 0.02-5 ng ml-1 using 100 microliters human plasma or urine. Inter-day variations of TA-0910 and TA-0910 acid-type assay in plasma were 3.5-15.5 and 1.8-9.4%, respectively. The variations of the assay in urine were the same as those in plasma. The recovery of TA-0910 and TA-0910 acid-type spiked in plasma or urine samples was approximately 100%. Furthermore, this method was applied to the determination of TA-0910 and TA-0910 acid-type in human plasma and urine samples, for the evaluation of the pharmacokinetics of TA-0910 in humans. From the results it was demonstrated that he developed RIA was useful for the determination of TA-0910 and TA-0910 acid-type in human plasma and urine, and was applicable to pharmacokinetic studies in humans.

Enzyme-linked immunosorbent assay for TA-2005-glucuronide in human plasma.

A sensitive enzyme-linked immunosorbent assay (ELISA) for TA-2005-glucuronide, a main metabolite of new adrenergic beta-receptor agonist TA-2005, has been investigated without prior deconjugation. Coupling of the hapten with bovine serum albumin (BSA) or beta-D-galactosidase was carried out by the N-hydroxysuccinimide ester method. An anti-TA-2005-glucuronide antiserum was obtained from guinea pig immunized with the hapten-BSA conjugate. The ELISA was based upon a competitive assay in which the separation of bound from free fraction was performed by the double antibody technique using rabbit anti guinea pig immunoglobulin antibody adsorbed to microtiter plates. A satisfactory standard curve for the ELISA of TA-2005-glucuronide was observed in the range of 30 pg-3 ng ml-1 using 25 microliters of human plasma. Inter-day and intra-assay variations were 7.0-17.5% and 1.0-11.7% respectively. The recoveries of TA-2005-glucuronide spiked to plasma samples were 95.5-120% (inter-assay) and 96.0-123.3% (intra-assay). The cross-reactivities of the prepared antiserum with the related compound of TA-2005-glucuronide were quite low though there was a considerable cross-reaction with TA-2005. However, TA-2005-glucuronide could be easily separated from TA-2005 by a simple pretreatment of the plasma sample with a C18 cartridge column. This method was applied to the determination of TA-2005-glucuronide in human plasma samples for the evaluation of the pharmacokinetics of TA-2005. From the results, it was demonstrated that the ELISA developed was useful for the determination of TA-2005-glucuronide in human plasma and that the method was applicable to pharmacokinetic studies in humans.

Radioimmunoassay for imidapril, a new angiotensin-converting enzyme inhibitor, and imidaprilat, its active metabolite, in human plasma and urine.

A radioimmunoassay (RIA) was investigated for the determination of imidapril and its active metabolite, imidaprilat, in human plasma and urine. Imidapril is a new angiotensin-converting enzyme inhibitor and an oral prodrug of imidaprilat. Imidapril was determined after conversion to imidaprilat with esterase. Antiserum was raised in rabbits against the p-amino derivative of imidaprilat conjugated to bovine serum albumin. Radioligand was prepared by iodination (125I) of the p-hydroxybenzoylamino derivative of imidaprilat. Cross-reactivities of anti-imidaprilat antiserum for imidapril, its metabolites and several cardiovascular drugs were low. The calibration range was 0.1-100 ng ml-1 using a 100 microliters of human plasma of urine. Intra- and inter-day variations of imidaprilat assay in plasma were 2.0-7.9 and 4.1-6.2%, respectively, and intra- and inter-day variations of imidapril assay in plasma were 5.4-10.7 and 7.9-18.1%, respectively. The variations of the assay in urine were a little smaller than those in plasma. The recovery of imidaprilat and imidapril spiked in plasma or urine samples was approximately 100%. A good correlation between RIA and high-performance liquid chromatograpy was observed for both plasma and urine samples. Furthermore, this method was applied to the determination of imidaprilat and imidapril in human plasma and urine samples, for the evaluation of the pharmacokinetics of imidapril in humans. From the results, it was demonstrated that the developed RIA was useful for the determination of imidaprilat and imidapril in human plasma and urine, and was applicable to pharmacokinetic studies in humans.

[Clinical significance of serum KL-6 in pulmonary sarcoidosis].

We investigated whether the level of serum KL-6 could be an activity marker for pulmonary sarcoidosis. In 33 patients with pulmonary sarcoidosis, the relationships between serum KL-6 levels and diagnostic imaging, serum angiotensin-converting enzyme (ACE) levels, serum lysozyme levels, steroid therapy, and prognosis were evaluated. There were no significant differences in the level of serum KL-6 when the patients were divided on the basis of radiographic findings, but the level of serum KL-6 was markedly elevated in some patients with stage-II pulmonary sarcoidosis. There was a significant correlation between serum KL-6 levels and the following two parameters: serum ACE and lysozyme levels. Among patients with a high initial level of serum KL-6, pulmonary sarcoidosis tended to become exacerbated within one year. Steroid therapy significantly decreased the level of serum KL-6, suggesting that the level of serum KL-6 could be an activity indicator for pulmonary sarcoidosis. Immunohistochemical staining by anti-KL-6 antibody revealed that KL-6 was localized in proliferating type-II alveolar epithelial cells.