Suguru Igarashi

Assessment of long-term function of heterotopic transplants of vitrified ovarian tissue in cynomolgus monkeys

BACKGROUND Ovarian tissue cryopreservation by rapid cooling (vitrification) is a convenient fertility preservation option. However, the progress of vitrified ovarian tissue after transplantation is not well understood in primates. METHODS For tissues from cynomolgus monkeys, we used closed straw vitrification and open cryosupport vitrification in which tissues are immersed directly into liquid nitrogen. Following warming, ovarian cortical pieces were autotransplanted and their function was monitored by computed tomography (CT), hormone assays and oocyte recovery, ICSI and embryo transfers (ETs). RESULTS Hormone cycles were restored in 6 of 7 animals in a mean of 126 days with no significant difference between the two vitrification regimens. The presence of new blood vessels supplying the grafted ovarian tissue was confirmed by contrast-enhanced CT. Oocyte retrieval from two monkeys after transplantation of the ovarian cortex vitrified by cryosupport vitrification yielded a total of nine oocytes of which six fertilized after ICSI, but ETs did not lead to any pregnancies. CONCLUSIONS This work shows that CT can give insight into ovarian function after heterotopic transplantation, and that heterotopic autografts of vitrified ovarian cortex can give rise to long-term ovarian function and embryos in a primate model. It remains to be established how outcomes following rapid vitrification compared with outcomes following conventional slow cooling procedures.

Risk factors for perioperative venous thromboembolism: A retrospective study in Japanese women with gynecologic diseases

BackgroundPatients with gynecologic cancer have a high risk of venous thromboembolism (VTE) like patients with other cancers. However, there is little information on risk factors for VTE during gynecologic surgery and no uniform preventive strategy. Our objectives were to identify risk factors for perioperative VTE in gynecologic patients and establish methods for prevention.MethodsWe analyzed 1,232 patients who underwent surgery at the Department of Obstetrics and Gynecology of St. Marianna University School of Medicine between January 2005 and June 2008. We investigated (1) risk factors for preoperative VTE, (2) use of an inferior vena cava (IVC) filter, and (3) risk factors for postoperative VTE.ResultsThere were 39 confirmed cases of perioperative VTE (3.17%), including 25 patients with preoperative VTE and 14 with postoperative VTE. Thirty-two patients had cancer and seven patients had benign diseases. Twenty-two of the 32 cancer patients (68.7%) had preoperative VTE, while postoperative VTE occurred in 10 cancer patients. Multivariate analysis indicated that ovarian cancer, tumor diameter ≥10 cm, and previous of VTE were independent risk factors for preoperative VTE. Among ovarian cancer patients, multivariate analysis showed that an age ≥50 years, the presence of heart disease, clear cell adenocarcinoma, and tumor diameter ≥20 cm were independent risk factors for preoperative VTE. The factors significantly related to preoperative VTE in patients with benign disease included previous VTE, age ≥55 years, tumor diameter ≥20 cm, and a history of allergic-immunologic disease. Thirteen of the 25 patients (52%) with preoperative VTE had an IVC filter inserted preoperatively. Postoperative screening (interview and D-dimer measurement) revealed VTE in 14/1,232 patients (1.14%). Multivariate analysis indicated that cancer surgery, a history of allergic-immunologic disease, and blood transfusion ≥2,000 ml were independent risk factors for postoperative VTE.ConclusionsPerioperative VTE is often fatal and preventive measures should be taken in the gynecologic field, especially when patients have the risk factors identified in this study. Since VTE is often present before surgery, preoperative screening is important and use of an IVC filter should be considered.

Heterotopic autotransplantation of ovarian cortex in cynomolgus monkeys.

In recent years, removal of ova or ovaries before chemotherapy or radiation therapy has been investigated in young female cancer patients to avoid the adverse effects of treatment. Orthotopic autotransplantation of ovarian cortex has advantages such as easy collection of ova and the possibility of spontaneous pregnancy. Although children have been born after successful orthotopic autotransplantation into the residual ovaries, some patients cannot undergo this procedure such as those who need bilateral ovariectomy or pelvic radiation therapy, therefore it is still necessary to investigate suitable heterotopic autotransplantation sites. The present study was performed in primates (cynomolgus monkeys) with the objective of determining the optimum site for heterotopic autotransplantation of ovarian cortex to enhance the clinical application of this method. The retroperitoneal iliac fossa and omentum were selected as sites for heterotopic autotransplantation. Two cynomolgus monkeys were subjected to laparotomy under anesthesia. After resection of the bilateral adnexae, the ovaries were cut into 0.5 cm cubes that were transplanted. Blood levels of follicle-stimulating hormone, luteinizing hormone, estradiol, and progesterone were monitored, and monkeys with a regular estrus cycle underwent superovulation and egg collection. In both monkeys studied, recovery of a regular estrus cycle was confirmed after heterotopic autotransplantation of ovarian tissue. Mll phase ova were successfully collected from tissues transplanted into the retroperitoneal iliac fossa or omentum. Development to the early blastocyst stage was confirmed after microfertilization. We established an appropriate method of heterotopic autotransplantation using ovarian cortex into the retroperitoneal iliac fossa or omentum in primates.

N-Butyl Cyanoacrylate Is Very Effective for Massive Haemorrhage during the Perinatal Period

Objective The liquid embolic agent n-butyl cyanoacrylate (NBCA) is a tissue adhesive used as an immediate and permanent embolic agent when mixed with oil-based contrast medium. In this study, the preservation of fertility with TAE using NBCA for massive haemorrhage during pregnancy or the peripartum period and the utility of this therapy were investigated. Methods Cases from January 2005 to October 2010 in which TAE was performed for massive haemorrhage in pregnant women, particularly during the peripartum period, were investigated. Results TAE was performed in 27 pregnant women. The embolic agent used was GS only in five cases, NBCA only in 19 cases, and additional embolization with NBCA when the effect with GS was insufficient in three cases, one each of abruptio placentae, cervical pregnancy, and uterine atony.A comparison of mean blood loss when each embolic agent was used for haemostasis showed a significant difference between cases in which GS only was used and cases in which NBCA only was used. In a comparison of mean transfusion volume, a significant difference was seen between cases in which both GS and NBCA were used and cases in which NBCA only was used. In a postoperative follow-up survey, menses resumed in eight patients, including four patients who later became pregnant and three who delivered. Conclusions TAE with NBCA, which has an embolic effect unrelated to clotting dysfunction for massive haemorrhage during the peripartum period, is a minimally invasive and very effective treatment method for patients with severe DIC.

Prenatal diagnosis of pyriform sinus fistula: case report and literature review

We describe a case of prenatal diagnosis of a pharyngeal cyst as a pyriform sinus fistula on the findings of ultrasonography and magnetic resonance imaging.

Ovarian Tissue Cryopreservation: Ovarian Cortical Tissue Vitrification

Vitrification has become common practice in cryopreservation of embryo, blastocyst, oocytes, and sperm. Freezing of ovarian tissue has been studied since the late 1990s, and in 2004, Donnez et al. reported that live births could be achieved with this method. Reports worldwide have indicated that slow freezing is a common method of cryopreservation. Successive reports have emerged since 2005 describing ovarian tissue cryopreservation by vitrification in animal experiments and using human tissue; however, there have been no reports to date in which vitrification was clinically applied and live births were successfully achieved. We have been conducting studies through pre-clinical tests using Cynomolgus monkey (Macaca fascicularis) since 2006, and as a result, at present in June, 2016, we have achieved ovarian tissue cryopreservation in 229 cases and 2 live births following thawed transplantation. This article outlines the research findings so far regarding ovarian tissue cryopreservation by vitrification; additionally, it describes our ovarian cortical tissue vitrification method.

Menopause-specific health literacy in Japanese women

OBJECTIVES This study proposed a method for assessing menopause-specific health literacy (knowledge and beliefs about menopausal symptoms which aid their recognition, assessment, and management) using a vignette methodology. STUDY DESIGN A cross-sectional web-based survey was conducted in September 2015 among Japanese women aged 30-59 years. Of 1236 women surveyed, 1196 eligible participants who were not under treatment for menopausal symptoms were included. MAIN OUTCOME MEASURES Participants were presented with a vignette describing a woman with menopausal symptoms and were then asked a series of questions to assess their recognition of menopausal symptoms, attitude, subjective norm, perceived behavior control, availability, and intention to seek medical care if they themselves had the problems described in the vignette. RESULTS The majority (87%) of participants correctly labelled the vignette as menopausal symptoms and 60% expressed an intention to seek medical care if they had the symptoms presented. Logistic regression showed that attitude, subjective norm, and perceived behavior control were significant predictors of the intention to seek medical care. A structural equation model depicting these relationships with intention to seek medical care revealed acceptable fit indices: goodness of fit index (GFI)=0.948, adjusted goodness of fit index (AGFI)=0.913, comparative fit index (CFI)=0.883, and root mean square error of approximation (RMSEA)=0.089. Subjective norm had the greatest direct effect on intention to seek medical care. CONCLUSIONS The assessment of menopause-specific health literacy may be useful for understanding why women hesitate to seek medical care for menopausal symptoms and for developing interventions to improve the coping behaviors of women with menopausal symptoms.

Oocyte retrieval after heterotopic transplantation of ovarian tissue cryopreserved by closed vitrification protocol

PurposeA device for closed vitrification was designed to reduce the risk of contamination and investigated on its efficacy for ovarian function recovery after cryopreservation and heterotopic transplantation.MethodsOvarian tissues from green fluorescence protein transgenic mice (10 GFP mice) were vitrified using the device, and warmed ovarian tissues were transplanted into the ovarian bursa region in wild-type female mice (6 mice). Fresh ovarian tissues were similarly transplanted as a control. After recovery of the estrous cycle, mice were mated with male mice. Ovarian tissues from six cynomolgus monkeys were vitrified and warmed with the device for autologous, heterotopic transplantation. Fresh tissue transplantation was not performed for the control. Ovarian function was examined by recovery of the hormonal cycle. Histological examination was conducted.ResultsThe number of live pups per recipient mouse was not significantly different after transplantation of fresh or vitrified-warmed ovarian tissue, although the pregnancy rate was reduced with vitrified tissues. The hormonal cycle was restored in 5/6 monkeys after heterotopic transplantation of vitrified-warmed ovarian tissue. Follicles were harvested at eight sites in the omentum and 13 sites in the mesosalpinx. In vitro maturation (IVM)/IVF produced embryo but did not develop.ConclusionsResumption of the hormonal cycles, follicle development, and oocyte retrieval from vitrified-warmed ovarian tissue transplants may indicate that the use of vitrification for ovarian tissue in a closed system has a potential of clinical application without the risk of contaminations. More detailed analyses of the effects of vitrification on ovarian tissue, such as gene expression patterns in oocytes and granulosa cells, may be needed for establishing a standard procedure for cryopreservation of ovarian tissues in human.

Anti-aging Effect on Skin of Autologous Transplantation of Tissue Fragments from Thawed Cryopreserved Ovaries

Transplanting the ovaries of young mice into menopausal mice has been shown to extend their lifespan, suggesting that the reproductive organs may play an important role in combating aging. Preventing skin aging is an extremely important matter with respect to maintaining quality of life, but little basic research has been car‐ ried out on this issue. The effects of treatment with cryopreserved ovarian tissue, tissue hormone therapy (THT), and hormone replacement therapy on inhibiting skin aging in experimental animals were investigated. The ef‐ fects on skin elasticity and body weight changes in 6-week-old mice resulting from the transplantation of cryo‐ preserved ovarian tissue were evaluated, as were the effects on skin of estrogen administration after bilateral oophorectomy or transplantation. Estrogen was secreted by mouse ovaries that had been frozen, thawed, and transplanted, and the estrus cycle was restored, but this was insufficient to have any effect on skin elasticity. After oophorectomy, the body weight of the mice increased, and their skin elasticity decreased. Estrogen admin‐ istration after these changes had occurred neither restored skin elasticity nor suppressed body weight gain. However, when estrogen was continuously administered from immediately after oophorectomy, skin elasticity decreased transiently and then improved. If techniques for THT using cryopreserved ovaries to maintain the blood estrogen concentration above a specific level can be established, this might help to prevent or improve the deterioration of skin appearance in young women who require oophorectomy due to gynecological disease and also in childhood, adolescent, and young adult cancer patients.

Peripartum heart failure caused by left ventricular diastolic dysfunction: a case report

Peripartum cardiomyopathy is a rare but potentially life‐threatening condition. The current definition of peripartum cardiomyopathy only includes patients with systolic dysfunction. We describe a 25‐year‐old nulligravid patient with heart failure, i.e. left ventricular diastolic dysfunction with preserved systolic dysfunction during the third trimester of pregnancy. She complained of dyspnea and was referred to our hospital at 31 weeks of gestation. The patient met the clinical criteria for peripartum cardiomyopathy with the exception of systolic dysfunction. Brain‐type natriuretic peptide levels peaked at 1447 pg/dL. The patient responded to therapy for heart failure and showed resolution of her diastolic dysfunction by 1 month postpartum. The case demonstrated the important role of diastolic dysfunction in peripartum heart failure and the possibility of clarifying the pathophysiology of peripartum cardiomyopathy by evaluating diastolic function. Further investigations are needed to provide evidence regarding the clinical role of diastolic dysfunction in peripartum heart failure.