The Yokohama system for reporting directly sampled endometrial cytology: The quest to develop a standardized terminology.

BACKGROUND: The main purpose of directly sampled endometrial cytology is to detect invasive endometrial malignancies. With this principle in mind, The Yokohama System (TYS) Working Group, composed of cytopathologists, surgical pathologists, and gynecologic oncologists met at the 2016 International Congress of Cytology, Yokohama, with the aim to publish a standardized reporting system inclusive of specific diagnostic categories and cytomorphologic criteria for uniform and reliable diagnosis of endometrial malignancies on directly sampled endometrial samples. METHODS: The diagnostic cytopathologic criteria previously published in the literature by the Japanese and Greek working group on endometrial cytology (Yanoh et al. [2012] Acta Cytol. 56:233; Margari et al. [2016] Diagn Cytopathol. 44:888-901) were critically reviewed with the aim of correlating the diagnostic classes to well defined risk categories for endometrial carcinoma (EC). Moreover, two classes of "atypical" endometrial cells were correlated respectively to a low- and high risk group. Some methodological suggestions for the application of ancillary special technologies to liquid based samples were also given. RESULTS: The TYS group conceived a new Bethesda-style classification for directly sampled endometrial cytology which correlates the cytologic diagnostic classes with definite risk categories. The cytomorphologic findings have been correlated to the molecular pathology of EC, also through the application of ancillary special techniques to liquid-based samples. CONCLUSIONS: The success of TYS will depend on the acceptance of TYS by all the relevant pathology and gynecologic oncology communities who, by their joint efforts, will adopt, critically evaluate, and optimize this method with the only aim of further improving the impact of endometrial cytology on patients' care.

Production of optically pure D-lactic acid from brown rice using metabolically engineered Lactobacillus plantarum.

Simultaneous saccharification and fermentation (SSF) of D-lactic acid was performed using brown rice as both a substrate and a nutrient source. An engineered Lactobacillus plantarum NCIMB 8826 strain, in which the ʟ-lactate dehydrogenase gene was disrupted, produced 97.7 g/L D-lactic acid from 20% (w/v) brown rice without any nutrient supplementation. However, a significant amount of glucose remained unconsumed and the yield of lactic acid was as low as 0.75 (g/g-glucose contained in brown rice). Interestingly, the glucose consumption was significantly improved by adapting L. plantarum cells to the low-pH condition during the early stage of SSF (8-17 h). As a result, 117.1 g/L D-lactic acid was produced with a high yield of 0.93 and an optical purity of 99.6% after 144 h of fermentation. SSF experiments were repeatedly performed for ten times and D-lactic acid was stably produced using recycled cells (118.4-129.8 g/L). On average, D-lactic acid was produced with a volumetric productivity of 2.18 g/L/h over 48 h.

Generic Selection Criteria for Safety and Patient Benefit [IV] - Physicochemical and pharmaceutical properties of brand-name and generic ketoprofen tapes.

The physicochemical and pharmaceutical properties (pH, peel force, water-vapor permeability, and stretchability) of brand-name and generic ketoprofen products were evaluated and compared. The pHs of Mohrus as a brand-name product and Teikoku as a generic product were low (about 4). Among the other generic products, Patell and Nichi-Iko had a pH of about 4.3 while Frestol, Raynanon, BMD, and Touchron showed a pH of 4.6-5.2, which was in the pH range of normal healthy skin (4.5-6.5). The adhesive force was high (≥ 1.38) for Mohrus as a brand-name product as well as for Teikoku and Patell as generic products, but it was low (≤ 0.57) for the other 5 generic products. The water-vapor permeabilities of Mohrus as a brand-name product and Teikoku and Patell as generic products were low, being less than 1/6 of those for the other 5 generic products. Among the 5 generic products, BMD showed the highest water-vapor permeability (1,330 g/m²), and the other products also showed a value ≥ 1,100 g/m². The elongatedness of Mohrus was the lowest (15.5 cm), and that of Raynanon was the highest (24.5 cm); the difference was 9 cm. In this study, the physiochemical and pharmaceutical properties of ketoprofen tapes were clarified, which will allow pharmacists to provide products according to the needs of each patient when a brand-name product is changed to a generic one.

Production of d-lactic acid from hardwood pulp by mechanical milling followed by simultaneous saccharification and fermentation using metabolically engineered Lactobacillus plantarum.

This study focused on the process development for the d-lactic acid production from cellulosic feedstocks using the Lactobacillus plantarum mutant, genetically modified to produce optically pure d-lactic acid from both glucose and xylose. The simultaneous saccharification and fermentation (SSF) using delignified hardwood pulp (5-15% loads) resulted in the lactic acid titers of 55.2-84.6g/L after 72h and increased productivities of 1.77-2.61g/L/h. To facilitate the enzymatic saccharification of high-load pulp at a fermentation temperature, short-term (⩽10min) pulverization of pulp was conducted, leading to a significantly improved saccharification with the suppressed formation of formic acid by-product. The short-term milling followed by SSF resulted in a lactic acid titer of 102.3g/L, an optical purity of 99.2%, and a yield of 0.879g/g-sugars without fed-batch process control. Therefore, the process presented here shows promise for the production of high-titer d-lactic acid using the L. plantarum mutant.

Identification of a predictive biomarker for the beneficial effect of keishibukuryogan, a kampo (Japanese traditional) medicine, on patients with climacteric syndrome.

Keishibukuryogan (KBG; Guizhi-Fuling-Wan in Chinese) is one of the Kampo (Japanese traditional) medicines used to treat patients with climacteric syndrome. KBG can be used by patients who cannot undergo hormone replacement therapy due to a history of breast cancer. We evaluated whether cytosine-adenine (CA) repeat polymorphism of the estrogen receptor ß gene can be a predictor of the beneficial effect of KBG on climacteric syndrome. We also investigated the relationship between CA repeat polymorphism, the patients' profiles, and the therapeutic effect. We found that CA was an SS, SL, or LL genotype according to the number of repeats. We studied 39 consecutive patients with climacteric disorders who took KBG for 12 weeks. The diagnosis of climacteric disorders was made on the basis of the Kupperman index. KBG significantly improved the patients' climacteric symptoms (i.e., vasomotor symptoms in the patients with the LL genotype and melancholia in the patients with the SL genotype). No relationship between the patients' profiles and CA repeat polymorphism was recognized. CA repeat polymorphism could thus be a potential biomarker to predict the efficacy of KBG in climacteric syndrome, and its use will help to reduce the cost of treating this syndrome by focusing the administration of KBG on those most likely to benefit from it.

Complicating preeclampsia as a predictor of poor survival of the fetus in complete hydatidiform mole coexistent with twin fetus.

OBJECTIVE: To examine whether preeclampsia is a predictive factor for fetal prognosis in complete hydatidiform mole coexistent with twin fetus (CHMCF). STUDY DESIGN: We performed a retrospective chart review for 17 cases of definitive CHMCF managed in our hospital between 1991 and 2011. RESULTS: Fifteen patients chose expectant management and the remaining 2 selected termination of the pregnancy. During expectant management 6 patients displayed hypertension with proteinuria, representing preeclampsia, by the 2nd trimester (11-24 weeks) and the other 9 did not (nonpreeclamptic). No babies from preeclamptic mothers survived, with 5 intrauterine fetal deaths at 16-29 weeks and 1 neonatal death at 22 weeks. By contrast, 5 babies from 9 nonpreeclamptic mothers (1 preterm delivery at 29 weeks and 4 term deliveries) survived, while 4 pregnancies were lost by spontaneous abortion at 11-19 weeks. Low-risk gestational trophoblastic neoplasia (GTN) eventually occurred in both preeclamptic (4 of 6) and nonpreeclamptic (4 of 11) cases. Complicating preeclampsia correlated significantly with fetal demise and an increasing trend in serum hCG level but not with postmolar GTN. CONCLUSION: Complicating preeclampsia predicts poor survival of the fetus, but not subsequent GTN, in CHMCF.

Molecular distinction of consecutive molar pregnancies.

BACKGROUND: It may be difficult to differentiate the consecutive occurrence of two independent molar pregnancies from gestational trophoblastic neoplasia after the initial molar pregnancy, especially when the interval between them is short. CASE: A 25-year-woman who had had a complete hydatidiform mole 6 months earlier presented with a 6-week history of secondary amenorrhea. Serum human chorionic gonadotropin had increased to 19,857 micro-international units/mL, with no gestational sac demonstrated. Dilation and curettage was performed. Pathologic examination identified a tiny amount of hydropic villi compatible with complete hydatidiform mole. Analysis of short tandem repeat polymorphisms revealed that the molar tissues of the first and second complete hydatidiform moles were of different genetic origin. The patient went into remission spontaneously without chemotherapy. CONCLUSION: Genetic profiling was useful to discriminate a recurrent mole from suspected gestational trophoblastic neoplasia.

Comparison of 2 commercially available human chorionic gonadotropin immunoassays used in the management of gestational trophoblastic neoplasia.

OBJECTIVE: To compare serum human chorionic gonadotropin (hCG) titers using 2 commercially available hCG immunoassays in patients with gestational trophoblastic neoplasia (GTN). STUDY DESIGN: A total of 213 serum samples from 39 patients with uneventful moles and 697 serum samples from 17 patients with low-risk and high-risk GTN were obtained and subsequently measured with both the hCG C-terminal (hCG-CTP) and DPC Immulite 2000 tests. RESULTS: In patients with uneventful moles and GTN, serum hCG levels recorded using the hCG-CTP and DPC Immulite 2000 tests correlated well (r2 = 0.936 and r2 = 0.958). However, the serum hCG titers measured using the DPC Immulite 2000 assay were approximately 2.5- and 2.7-fold higher than those measured with the hCG-CTP test (p < 0.0001) when lower titers of hCG (< 40 mIU/mL) were tested. In addition, 3 and 1 patient, respectivelyl, with uneventful mole and GTN obtained positive results with the DPC Immulite 2000 test (3.0-4.2 mIU/mL) after demonstrating undetectable hCG levels with the hCG-CTP test (< 1.0 mIU/mL). CONCLUSION: At the lower levels of hCG, a few discrepancies between hCG titers measured using the 2 commercial immunoassays arose and may be attributed to the existence of hCG-related molecules. However, these hCG metabolites disappeared rapidly, the clinical importance of which remains unclear.

Recent changes of the incidence of complete and partial mole in Chiba prefecture.

AIMS: To study the changes of the incidence of complete mole (CM) and partial mole (PM) by 10-year age groups in Chiba Prefecture. METHODS: All women registered as CM and PMs in Chiba Prefecture during these 18 years were included in this study. The diagnosis of CM and PM was based on the macroscopic and/or microscopic findings. The annual numbers of pregnancy were obtained from the Division of Statistics in Chiba Prefecture Government. RESULTS: The incidence of CM at the upper and lower extremes of maternal age is higher than that of PM. The incidence of CM has decreased constantly at all maternal ages and significantly decreased in women of middle reproductive age (20-39 years old) since 1991, while that of PM has stayed constant during these 18 years. CONCLUSIONS: The incidence of CM and PM in Chiba Prefecture has become as low as that in Europe or the USA. These recent changes suggest that Japanese women may have lost the increased risk to ovulate a nuclear or inactive oocytes, or the differential diagnosis between CM and PM may be obscured with the macroscopic and/or microscopic findings.

Ovarian hyperstimulation caused by gonadotroph cell adenoma: a case report and review of the literature.

OBJECTIVE: We present a case of spontaneous ovarian hyperstimulation caused by pituitary gonadotroph macroadenoma, and include a review of the literature. CASE REPORT: A 27-year-old woman presented with irregular menstruation and bilateral multicystic enlargement of the ovaries. Serum estradiol (E(2)) levels were marginally elevated for the follicular phase but within the physiological range. Serum luteinizing hormone (LH) was extremely low, follicle-stimulating hormone (FSH) was normal, and prolactin (PRL) was high. Magnetic resonance imaging disclosed a pituitary macroadenoma. Immunohistochemical examination of the surgically removed adenoma showed intense reactivity for FSH and LH. After the operation, E(2), LH and PRL levels were normalized, the ovaries returned to a normal morphology, and regular menstrual cycles were resumed. CONCLUSION: A review of the literature showed that ovarian hyperstimulation caused by pituitary gonadotroph adenoma is not always accompanied by elevated FSH levels. High PRL and E(2) and low LH were reported in the majority of the cases, but E(2) may stay within the range observed in normal menstrual cycles.

Differential in vivo binding dynamics of somatic and oocyte-specific linker histones in oocytes and during ES cell nuclear transfer.

The embryonic genome is formed by fusion of a maternal and a paternal genome. To accommodate the resulting diploid genome in the fertilized oocyte dramatic global genome reorganizations must occur. The higher order structure of chromatin in vivo is critically dependent on architectural chromatin proteins, with the family of linker histone proteins among the most critical structural determinants. Although somatic cells contain numerous linker histone variants, only one, H1FOO, is present in mouse oocytes. Upon fertilization H1FOO rapidly populates the introduced paternal genome and replaces sperm-specific histone-like proteins. The same dynamic replacement occurs upon introduction of a nucleus during somatic cell nuclear transfer. To understand the molecular basis of this dynamic histone replacement process, we compared the localization and binding dynamics of somatic H1 and oocyte-specific H1FOO and identified the molecular determinants of binding to either oocyte or somatic chromatin in living cells. We find that although both histones associate readily with chromatin in nuclei of somatic cells, only H1FOO is capable of correct chromatin association in the germinal vesicle stage oocyte nuclei. This specificity is generated by the N-terminal and globular domains of H1FOO. Measurement of in vivo binding properties of the H1 variants suggest that H1FOO binds chromatin more tightly than somatic linker histones. We provide evidence that both the binding properties of linker histones as well as additional, active processes contribute to the replacement of somatic histones with H1FOO during nuclear transfer. These results provide the first mechanistic insights into the crucial step of linker histone replacement as it occurs during fertilization and somatic cell nuclear transfer.

Genetic origin and imprinting in hydatidiform moles. Comparison between DNA polymorphism analysis and immunoreactivity of p57KIP2.

OBJECTIVE: To evaluate whether p57KIP2 expression is concordant with the result of DNA polymorphism analysis in molar pregnancy. STUDY DESIGN: Eleven molar pregnancies diagnosed by pathologic findings between October 2002 and April 2004 were studied. Histopathologic diagnosis, DNA polymorphism analysis and p57KIP2 immunohistochemistry were investigated. RESULTS: DNA polymorphism analysis identified 3 biparental conceptuses as well as 4 dispermic androgenetic complete moles (CMs) and 4 suggestive monospermic CMs. Distinctly positive nuclear immunoreactivity of p57KIP2 was observed in a significant proportion of the villous cytotrophoblast and mesenchyme (30-60% of cells positive) in 3 cases of biparental conceptuses proven by DNA polymorphism. In contrast, p57KIP2 expression was negative (< 5% positive cells) in either the villous cytotrophoblast or mesenchyme in 8 cases of androgenetic conceptuses proven by DNA polymorphism. In all 11, p57KIP2 immunostaining was observed in the nuclei of extravillous trophoblasts that served as internal positive controls. CONCLUSION: Negative p57KIP2 immunoreactivity (paternally imprinted, maternally expressed gene) was in perfect concordance with the androgenetic origin of molar pregnancies proven by DNA polymorphism. The results suggest that p57KIP2 immunoreactivity, which can be performed in routine pathologic examinations, is a promising ancillary diagnostic tool to differentiate androgenetic CM from biparental conceptuses.

H1FOO is coupled to the initiation of oocytic growth.

We previously reported the discovery of a novel mammalian H1 linker histone termed H1FOO (formerly H1OO), a replacement H1, the expression of which is restricted to the growing/ maturing oocyte and to the zygote. The significance of this pre-embryonic H1 draws on its substantial orthologous conservation, singular structural attributes, selectivity for the germ cell lineage, prolonged nucleosomal residence, and apparent predominance among germ cell H1s. Herein, we report that the intronic, single-copy, five-exon (> or =5301 base pair) H1foo gene maps to chromosome 6 and that the corresponding primary H1foo transcript gives rise to two distinct, alternatively spliced mRNA species (H1foo(alpha) and H1foo(beta)). The expression of the oocytic H1FOO transcript and protein proved temporally coupled to the recruitment of resting primordial follicles into a developing primary follicular cohort and thus to the critical transition marking the onset of oocytic growth. The corresponding potential protein isoforms (H1FOO(alpha) and H1FOO(beta)), both nuclear localization sequence-endowed but export consensus sequence-free and possessing a significant net positive charge, localized primarily to perinucleolar heterochromatin in the oocytic germinal vesicle. Further investigation will be required to define the functional role of the H1FOO protein in the ordering of the chromatin of early mammalian development as well as its potential role in defining the primordial-to-primary follicle transition.

Effects of follicle size and oocyte maturation conditions on maternal messenger RNA regulation and gene expression in rhesus monkey oocytes and embryos.

The relationship between alterations in gene expression and differences in developmental potential in primate oocytes and embryos was examined. Oocytes from 3 sources were used for these studies: 1) in vivo-matured oocytes from monkeys stimulated with FSH and hCG, 2) in vitro-matured oocytes from large follicles of monkeys primed with FSH, and 3) in vitro-matured oocytes from small follicles from nonstimulated (NS) monkeys. Following in vitro fertilization, embryos from these oocytes displayed high, moderate, and low developmental competence, respectively. Oocytes from NS females displayed aberrant accumulation of a number of maternal mRNAs, followed by precocious loss of many maternal mRNAs by the 2-cell stage. Embryos from NS oocytes displayed alterations in expression of key transcription factors after the 8-cell stage. Oocytes and embryos from FSH-stimulated females also displayed alterations in gene expression relative to hCG-stimulated females, but these alterations were much less severe than those observed for NS oocytes and embryos. Our data are consistent with the hypothesis that continued development and maturation of the oocyte within the ovarian follicle in vivo facilitates the production of oocytes of the highest developmental potential, and that in vitro conditions may not support this process as effectively due to differences in the extracellular milieu. These observations are relevant to understanding the role of the in vivo environment on oocyte maturation, and the potential effects of in vitro maturation on human assisted reproduction methods.

H1oo: a pre-embryonic H1 linker histone in search of a function.

The mouse oocyte-specific linker histone H1oo (1) constitutes a novel mammalian homologue of the oocyte-specific linker histone B4 of the frog and of the cs-H1 linker histone of the sea urchin; (2) is expressed as early as the germinal vesicle (PI) stage oocyte, persisting into the MII stage oocyte, the oocytic polar bodies, and the 2-cell embryo, extinction becoming apparent at the 4-8 cell embryonic stage; and (3) may play a key role in the control of gene expression during oogenesis and early embryogenesis, presumably through the perturbation of chromatin structure.