A multi-centre, retrospective case series of oocyte cryopreservation in unmarried women diagnosed with haematological malignancies.

STUDY QUESTION: Is oocyte cryopreservation an applicable option for fertility preservation in unmarried patients with haematological malignancies? SUMMARY ANSWER: Oocyte cryopreservation via the vitrification method is accessible and may be considered an option for fertility preservation in unmarried patients with haematological malignancies. WHAT IS KNOWN ALREADY: Haematological malignancies are most commonly observed amongst adolescent and young adult women. Although the survival rate and life expectancy of those with haematological malignancies have improved, chemotherapy and radiotherapy may impair their reproductive potential. Oocyte cryopreservation is thus an ideal option to preserve their fertility. STUDY DESIGN SIZE DURATION: This study retrospectively evaluated 193 unmarried patients (age: 26.2 ± 0.4 years) with haematological malignancies, who consulted for oocyte cryopreservation across 20 different fertility centres in Japan between February 2007 and January 2015. The primary outcome measures were the oocyte retrievals and oocyte cryopreservation outcomes. The secondary outcome measures were the outcomes following oocyte warming for IVF. PARTICIPANTS/MATERIALS SETTING METHODS: The patients had commenced ovarian stimulation cycles via antagonist, agonist, natural and minimal methods for oocyte retrievals, defined according to the treatment strategy of each respective fertility centre. A vitrification method using the Cryotop safety kit was used for oocyte cryopreservation. ICSIs were used for insemination of warmed oocytes. The endometrial preparation method for embryo transfer was hormonal replacement therapy, except in the case of a patient who underwent a spontaneous ovulatory cycle. MAIN RESULTS AND THE ROLE OF CHANCE: Among 193 patients, acute myeloid leukaemia (n = 45, 23.3%) was most common, followed by acute lymphoid leukaemia (n = 38, 19.7%) and Hodgkin's lymphoma (n = 30, 15.5%). In total, 162 patients (83.9%) underwent oocyte retrieval, and oocytes were successfully cryopreserved for 155 patients (80.3%). The mean number of oocyte retrieval cycles and cryopreserved oocytes were 1.7 ± 0.2 and 6.3 ± 0.4, respectively. As of December 2019, 14 patients (9.2%) had requested oocyte warming for IVF. The survival rate of oocytes after vitrification-warming was 85.2% (75/88). The rates of fertilisation and embryo development were 80.0% (60/75) and 46.7% (28/60), respectively. Ten patients (71.4%) had successful embryo transfers, and seven live births (50.0%) were achieved. LIMITATIONS REASONS FOR CAUTION: This study was limited by its retrospective nature. Additionally, there remains an insufficient number of cases regarding the warming of vitrified oocytes to reliably conclude whether oocyte cryopreservation is effective for patients with haematological malignancies. Further long-term follow-up study is required. WIDER IMPLICATIONS OF THE FINDINGS: Oocyte retrieval and oocyte cryopreservation were accessible for patients with haematological malignancies; however, the number of oocyte retrievals may have been limited due to the initiation of cancer treatments. Acceptable embryonic and pregnancy outcomes could be achieved following oocyte warming; therefore, our results suggest that oocyte cryopreservation can be considered an option for fertility preservation in patients with haematological malignancies. STUDY FUNDING/COMPETING INTERESTS: This research received no specific grant from any funding agency in the public, commercial or not-for-profit sectors. The authors declare no conflict of interest. TRIAL REGISTRATION NUMBER: N/A.

CodY is a nutritional repressor of flagellar gene expression in Bacillus subtilis.

Expression of the sigma(D)-dependent flagellin gene, hag, is repressed by the CodY protein in nutrient-rich environments. Analysis of a codY mutant bearing a hag-lacZ reporter suggests that the availability of amino acids in the environment is the specific signal that triggers this repression. Further, hag-lacZ expression appears to be sensitive to intracellular GTP levels, as demonstrated by increased expression upon addition of decoyinine. This result is consistent with the postulate that the availability of amino acids in the environment effects intracellular GTP levels through the stringent response. However, the levels of hag-lacZ measured upon the addition of subsets of amino acids suggest an additional mechanism(s). CodY is a DNA binding protein that could repress flagellin expression directly by binding to the hag promoter region, or indirectly by binding to the fla/che promoter region that governs expression of the sigma(D) transcriptional activator required for hag gene expression. Using an electrophoretic mobility shift assay, we have demonstrated that purified CodY protein binds specifically to both the hag and fla/che promoter fragments. Additionally, CodY acts as a nutritional repressor of transcription from the fla/che promoter region that contains two functional promoters. CodY binds to both the sigma(D)- and sigma(A)-dependent promoters in this region, as demonstrated by DNase I footprint analyses. Footprint analyses of the hag gene demonstrated that CodY binds downstream of its sigma(D)-dependent promoter. Taken together, these results identify new members of the CodY regulon that encode motility functions in Bacillus subtilis and are controlled by the sigma(D) alternate sigma factor.

MR imaging of the fetus by a HASTE sequence.

OBJECTIVE: The value of a half-Fourier acquisition single-shot turbo spin-echo (HASTE) sequence, in which high-resolution heavily T2-weighted images can be obtained within 2 sec, was evaluated in the imaging of the fetus during the second or third trimester. MATERIALS AND METHODS: Eighteen women with complicated pregnancies as revealed on a sonogram during the second and third trimesters (16-36 weeks' gestation) were studied with a 1.5-T superconductive MR imaging unit that used a body phased-array coil. After informed consent, T1-weighted fast low-angle shot images, T2-weighted turbo spin-echo images, and HASTE images were obtained without any premedication. Images were analyzed with regard to image quality, degree of blurring, visualization of the normal fetal organs, and visualization of fetal and maternal abnormalities. RESULTS: On HASTE sequences, visualization of the fetal brain, visceral organs (lung, heart, liver, kidney, and bladder), extremities, and umbilical cord were significantly better than on fast low-angle shot or turbo spin-echo sequences (p < .01). In the brain, the white matter-gray matter distinction, gyrus formation, and myelination of the brain were clearly revealed by the HASTE sequence. Pathologic processes including fetal abnormalities (anomalies of the central nervous systems [n = 5], placenta previa [n = 1], and transverse lie in the third trimester [n = 1]) and maternal abnormalities (leiomyoma [n = 5], ovarian tumors [n = 3], and hydronephrosis [n = 1]) were clearly seen on HASTE imaging. The peak specific absorption rate for RF exposure in these studies was less than 1.5 W/kg. CONCLUSION: In situations when sonography is suggestive but not definitive, MR imaging with a HASTE sequence allows clear fetal imaging with high T2-weighted contrast.

Adnexal masses: accuracy of characterization with transvaginal US and precontrast and postcontrast MR imaging.

PURPOSE: To determine the accuracy of transvaginal ultrasound (TVUS) and of precontrast and contrast material-enhanced magnetic resonance (MR) imaging in the differentiation of adnexal masses. MATERIALS AND METHODS: Five blinded readers analyzed images of 80 masses in 72 patients. MR and TVUS images were interpreted in separate sessions. Findings were confirmed at surgery or laparoscopy. RESULTS: Higher diagnostic accuracy was attained with MR imaging in mature cystic teratomas and endometriomas. However, better accuracy was achieved with contrast-enhanced MR imaging and TVUS in simple cysts, cystadenomas, and malignant tumors because internal details could be visualized. Receiver operating characteristic study indicated that observer confidence was significantly higher with contrast-enhanced MR imaging than with precontrast MR imaging (P = .011) or TVUS (P = .002) in the differentiation of benign and malignant masses. CONCLUSION: Contrast-enhanced MR imaging is superior to precontrast MR and TVUS imaging in the characterization and differentiation of adnexal masses.

Effects of progesterone and anti-progesterone RU486 on ovarian 3 beta-hydroxysteroid dehydrogenase activity during ovulation in the gonadotrophin-primed immature rat.

The effects of progesterone and RU486, a synthetic anti-progesterone, on ovarian 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activity, a key enzyme of progesterone production, were studied during ovulation in immature 22-day-old rats primed with pregnant mares' serum gonadotrophin (PMSG) and human chorionic gonadotrophin (hCG). Ovarian 3 beta-HSD activities had increased significantly 4 h after hCG injection. These increases were inhibited at 4 and 6 h after hCG when 20 mg RU486 kg-1 was administered 2 h before hCG. However, RU486 had no influence on the activity of 3 beta-HSD when administered at the same time as hCG injection. A histochemical study revealed that 3 beta-HSD activities in the granulosa cell layer, but not in the theca cell layer, were inhibited when RU486 was given 2 h before hCG. Serum progesterone concentrations, but not oestradiol concentrations, were significantly suppressed by RU486 treatment 4 and 6 h after hCG. The effect of progesterone on ovarian 3 beta-HSD activity was tested by administering graded doses of progesterone exogenously to rats 2 h before hCG injection. Ovarian 3 beta-HSD activity was increased in a dose-dependent manner, and more than 20 mg progesterone kg-1 significantly stimulated the activity. Although 10 mg progesterone kg-1 did not stimulate ovarian 3 beta-HSD activities, the RU486-inhibited activities were recovered by the concomitant administration of 10 mg progesterone kg-1 with RU486. These results indicate that ovarian 3 beta-HSD activity depends on progesterone concentrations, and suggest an autocrine regulation of progesterone production during ovulation in immature rat ovaries stimulated with PMSG and hCG.

The relationship between ovarian progesterone and proteolytic enzyme activity during ovulation in the gonadotropin-treated immature rat.

To clarify the mechanisms by which progesterone acts as a mediator in the ovulatory process, ovulation rate and proteolytic enzyme activities were investigated in immature 22-day-old rats treated with PMSG/hCG, RU486 (10 mg/kg), synthetic antiprogesterone, and RU486 (10 mg/kg) + progesterone (10 mg/kg). The number of ova was significantly decreased when RU486 (10 mg/kg) was given from 2 h before to 4 h after the hCG injection. In addition, its inhibitory action on ovulation was reversed by exogenous progesterone (10 mg/kg) at 2 or 4 h after the hCG injection. Serum progesterone and estradiol concentrations in the rats treated with RU486 did not show any significant differences compared to controls. The proteolytic enzyme activities were measured by using the synthetic substrates alpha-N-benzoyl-DL-Arg-beta naphthylamide (BANA) and dinitrophenyl peptide (DNP). Activities were significantly increased after hCG injection in the control group during 8-9 h for BANA hydrolase and 7-10 h for DNP peptidase. The preovulatory increase of these activities was totally suppressed by RU486 with hCG. After administration of progesterone (10 mg/kg) following hCG and RU486 injection, the elevation of proteolytic, enzyme activities in the preovulatory phase was effectively reversed, and levels became similar to those in the control group. These results suggest that progesterone plays an indispensable role in the first 4 h of the ovulatory process by regulating proteolytic enzyme activities.

[The effect of progesterone and RU486 on progesterone production in the ovulatory process of rats].

To clarify the autoregulatory mechanism of progesterone (P) production in the ovulatory process, we examined the ovarian concentration of P 46 hrs after PMSG and the effects of P and RU486 (RU) injected 4-2 hrs before hCG administration on the serum concentrations of P and estradiol (E2), and ovarian 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activities in PMSG/hCG treated immature rats. The effect of RU on the number of ovulated ova was also studied. The ovarian concentration of P 46 hrs after PMSG was 0.96 +/- 0.03,ng/mg protein (mean +/- SEM). When P (100,mg/kg) was injected 2 hrs before hCG, ovarian 3 beta-HSD activities had significantly increased by 4 hrs after hCG. However, P at a dosage of 10 and 20,mg/kg had no effect on ovarian 3 beta-HSD activities. The administration of RU (20,mg/kg) 2 hrs before hCG significantly inhibited ovarian 3 beta-HSD activities measured 4 and 6 hrs after hCG (p less than 0.01 and p less than 0.05, respectively). In addition, the serum P concentration 4 hrs after hCG was significantly lower than that of the control (p less than 0.01). However, RU (20 mg/kg) in concomitant with hCG had no effect on ovarian 3 beta-HSD activities within 6 hrs after hCG. The suppression of ovarian 3 beta-HSD activities by RU was the concomitant reversed by the concomitant treatment with P (10 mg/kg). RU (10, 20, or 40 mg/kg) injected 2 hrs before hCG significantly reduced the number of ovulated ova (p less than 0.01, p less than 0.01 and p less than 0.01, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

[Midluteal progesterone/estradiol ratio as an indicator of pregnancy potential].

In an effort to determine the reliability of the midluteal progesterone(P)/estradiol(E2) ratio as an index of the potential for conception, we measured the midluteal P and E2 levels in 76 infertile women who had been treated at our infertility clinic. This parameter in conception cycles was compared with that in non-conception cycles. Eighty cycles of the 76 women were classified into two groups, depending upon whether pregnancy occurred or not. Group 1 and group 2 were composed of 31 conception cycles and 49 drug-induced cycles, respectively. Midluteal concentrations of P and E2 did not show any significant differences between the two groups. The P/E2 ratios were 106.4 +/- 71.3 (mean +/- SD) and 71.5 +/- 44.16, respectively. This difference was statistically significant (p less than 0.05). Discriminant function analysis showed that the smallest probability of misclassification between the two groups decreased from 44% when using P only to 36% when using the combined P and E2. The following equation: Y = 0.0727X1 - 0.00456X2 - 0.130 was obtained (when X1 = P, X2 = E2). These results suggest that the midluteal P/E2 ratio gives clinicians the best indication of luteal function for the achievement of pregnancy.

[Role of progesterone in the ovulatory process of PMSG/hCG treated immature female rats].

To determine exactly when progesterone (P) acts as a most important mediator in the ovulatory process, a dose of 10mg/kg of RU486, an antiprogesterone, was administered to PMSG/hCG treated immature female rats (22 days old) at 0 (RU0 groups), 2 (RU2 group), 4,6,8 or 10 hours after an hCG injection, respectively. In the RU0 groups, the ovulatory effect of P was investigated at 0.2 (P2 group), 4 (P4 group), 6 or 8 hours after the hCG injection. Ovulation rates were calculated 24 hours after the hCG injections. Serum P and estradiol (E2) concentrations in the RU0 groups, the P2 group and the control rats (C group), were determined. The results were as follows. 1) The numbers of ova in the RU0, RU2 and RU4 group rats were significantly (p less than 0.01, p less than 0.01 and p less than 0.01) lower than that in the C group rats. 2) In the P2 and P4 groups, the numbers of ova were significantly (p less than 0.01 and p less than 0.01) increased compared with the RU0 group and returned to the control range. 3) In the RU0 group, the serum P and E2 concentrations within 8 hours after an hCG injection did not show any significant differences compared with the C group. In contrast, in the P2 group, the P concentrations at 4 and 6 hours after hCG increased significantly (p less than 0.01 and p less than 0.01) compared to the corresponding RU0 group. These results clarified an indispensable role of P in the ovulatory process within 6 hours after an hCG injection.