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Background/Purpose: Abnormal folliculogenesis is one of the cardinal presentations of polycystic ovarian syndrome (PCOS) and permeability of follicular wall has been proposed to be involved in the normal follicular growth. However, whether or not there is a change in intrafollicular permeability underlies PCOS is unknown. Methods: This was a tertiary center-based case–control study. From 2014 to 2015, thirteen patients with PCOS who underwent in vitro fertilization-embryo transfer (IVF-ET) were enrolled. Eleven normo-ovulatory patients who underwent IVF-ET due to male factor and/or tubal factor infertility were enrolled as the control group. The influence of ovarian follicular fluid (FF) on endothelial cell permeability was evaluated using a human umbilical vein endothelial cell monolayer permeability assay. The intrafollicular expression profiles of angiogenesis-related proteins were analyzed using a Human Angiogenesis Protein Array Kit. Results: The FF from PCOS patients caused significantly poorer endothelial cell permeability comparing with the effect of FF from the control group (46% ± 12% vs. 58% ± 9%, P = 0.023). Among the 55 angiogenesis-related proteins tested, there was a significantly higher level of intrafollicular platelet factor 4 (PF4) and PF4/IL-8 complex in the PCOS group (p = 0.004). The anti-permeability effect of PF4 was related to the decrease in the intercellular gaps and antagonistic binding with IL-8. Conclusion: Our study provides the first evidence of the pathophysiologic contribution of the well-known angiostatic protein, PF4, on human reproductive biology. The increase of the intrafollicular PF4 and its anti-permeability effect might affect the formation of FF and folliculogenesis in PCOS. Keywords: Angiogenesis, Folliculogenesis, Permeability, Platelet factor 4, Polycystic ovarian syndrome

Objective Obesity has been reported to have a modulatory effect on the ovulatory functions of patients with polycystic ovary syndrome. The role of adipokines in this obesity-associated ovulatory disturbance has not been extensively explored. In this study, the relationships between obesity, adipokine production from visceral fat, and ovarian folliculogenesis were explored in a mice model of induced obesity. Methods Obesity was induced in female C57BL/6 mice fed ad libitum with high-fat feed and fructose water for 4 weeks. Follicular developments in the ovaries were assessed by histopathology in these diet-induced obese mice. Changes in adipokine expression in the peri-ovarian adipose tissues were screened with an adipokine array. The adipokine with the most significant increase over time was identified. The functions of the adipokine in angiogenic processes were evaluated in a cell model of endothelial proliferation. The in vivo effects of neutralizing this adipokine using specific antibodies were assessed in the same obesity model. Results A high-fat and fructose diet induced an accumulation of early ovarian follicles and a reduction in mature follicles and corpus lutea. The number of microvessels in the early follicles also decreased. The adipokine protein array of the peri-ovarian adipose tissues identified a progressive increase in IL-10 expression with the duration of the obesogenic diet. In vitro experiments in the endothelial cell model confirmed IL-10 as a disrupter of VEGF-induced angiogenesis. Administration of anti-IL-10 antibodies prevented the histopathological changes induced by the obesogenic diet and further highlighted the role of IL-10 in disrupting folliculogenesis. Conclusions Obesity may disrupt normal folliculogenesis through increased production of IL-10 in visceral fats. This relationship may help clarify the reported association between obesity and ovulatory dysfunction, which has been found in patients with polycystic ovary syndrome. However, the duration of this study was short, which limited conclusions on the long-term reproductive outcomes., Highlights • Obesity increases IL-10 expression in visceral adipose. • IL-10 disrupts VEGF-induced angiogenesis in an endothelial cell model. • Disrupted angiogenesis is associated with disturbed folliculogenesis. • Anti-IL-10 antibody prevents the altered folliculogenesis induced by obesity. • Abnormal production of IL-10 may be a cause of dysovulation in obese individuals.

In order to enhance anaerobic hydrogen production from yeast waste, a series of 120-mL batch co-cultures of Clostridium beijerinckii L9, Clostridium butyricum M1, and Bacillus thermoamylovorans B5 under mesophilic conditions were established according to full factorial design (FFD) and mixture design (MD). The experimental results were subjected to multivariate and response surface analyses to determine the relationships between bacteria converting yeast waste into hydrogen. The results indicated clearly that C. beijerinckii L9 and C. butyricum M1 had significant potential to convert yeast waste into hydrogen. There was no significant hydrogen generation when B. thermoamylovorancs B5 alone was cultured with yeast waste. However, B. thermoamylovorancs B5 could significantly shorten the co-culture’s hydrogen-producing lag phase. Response surface analyses demonstrate that B. thermoamylovorancs B5 can stimulate the specific hydrogen production rate of C. beijerinckii L9 and C. butyricum M1, greater in the case of the former than of the latter. An ultimate hydrogen yield of 46 mL H 2 /g COD added yeast waste was obtained with an optimal volumetric ratio C. beijerinckii L9: C. butyricum M1: B. thermoamylovoranc B5 of 8.9:4.8:10.3. Highly reproducible co-culture results confirm that FFD and MD, via response surface analysis, are applicable to assess the roles of the individual microorganisms in the defined co-culture.

Purpose Hepatitis B virus (HBV) reactivation can occur after radiotherapy (RT) for hepatobiliary malignancies. Our previous in vitro culture study identified interleukin-6 (IL-6) as the main bystander mediator of RT-induced HBV replication. We attempted to examine the molecular mechanism in HBV-transgenic mice. Methods and materials HBV transgenic mice were treated with whole liver RT (4 Gy daily for 5 days) with or without administration of IL-6 (400 ng twice daily for 15 days). The serum level of HBV DNA was measured using real-time polymerase chain reaction, and the IL-6 concentration was measured using enzyme-linked immunosorbent assay. The intensity of immunostaining with antibodies to HBV core protein and phosphorylated signal transducer and activator of transcription (STAT)3 in the mouse liver was qualitatively analyzed. HepG2.2.15 cells (a human hepatoblastoma cell line that persistently produces HBV DNA) were used to investigate the molecular role of IL-6 plus RT in HBV reactivation. Results HBV reactivation was induced in vivo with IL-6 plus RT (5.58-fold) compared with RT alone (1.31-fold, p = .005), IL-6 alone (1.31-fold, p = .005), or sham treatment (1.22-fold, p = .004). HBV core protein staining confirmed augmentation of intrahepatic HBV replication. IL-6 plus RT-induced HBV DNA replication in HepG2.2.15 cells was suppressed by the STAT3 inhibitor AG490 and by transfection with dominant-negative STAT3 plasmid. Phosphorylated STAT3 staining was strongest in liver tissue from mice treated with IL-6 plus RT. The mobility shift assay demonstrated that reactivation was mediated through the interaction of phosphorylated STAT3/hepatocyte nuclear factor-3 complex with HBV enhancer 1. Conclusion RT to the liver and longer sustained IL-6 induced HBV reactivation through the STAT3 signal transduction pathway.

Summary Objective The human endometrium manifests different and distinct morphologies and physiologies during the different phases of the menstrual cycle. We aimed to determine which candidate genes demonstrate differential expression patterns in the endometrium during different phases of the menstrual cycle. Materials and Methods Using differential display reverse transcription polymerase chain reaction to compare day 5 and day 18 human glandular endometrium obtained by laser capture microdissection, we identified a specific gene, NUDT9 (nucleoside diphosphate-linked moiety X motif 9). NUDT9 is known to function as a highly specific adenosine diphosphate ribose pyrophosphatase and has been mapped to chromosome 4q22.1. It gives rise to two alternatively spliced messenger RNAs, NUDT9α and NUDT9β, encoding a member of the Nudix hydrolase family. In this study, we purified NUDT9 protein and produced an antibody, which we then used for immunohistochemical studies. Results Using this anti-NUDT9 antibody, we successfully demonstrated that NUDT9 protein was differentially expressed in endometrial glandular cells at different phases of the menstrual cycle. NUDT9 was also found to be expressed more prominently in the epithelial glandular component than in the stromal component of human endometrial carcinomas. Conclusion We suggest that NUDT9 may be involved in the regulation of the menstrual cycle and may be related to the proliferation of glandular cells in the human endometrium.

By using brewery yeast waste and microflora from rice straw compost, an anaerobic semi-solid bio-hydrogen-producing system has been established. For the purpose of industrialization, the major players of both aerobic and anaerobic bacterial strains in the system were isolated and their combination for an effective production of bio-hydrogen and other bio-fuels was examined in this study. The phylogenetic analysis found that four anaerobic isolates (Clostridium beijerinckii L9, Clostridium diolis Z2, Clostridium roseum Z5-1, and C. roseum W8) were highly related with each other and belongs to the cluster I clostridia family, the family that many of solvent-producing strains included. On the other hand, one of the aerobic isolates, the Bacillus thermoamylovorans strain I, shown multiple extracellular enzyme activities including lipase, protease, α-amylase, pectinase and cellulase, was suggested as a good partner for creating an anaerobic environment and pre-saccharification of substrate for those co-cultured solventogenic clostridial strain. Among these clostridial strains, though C. beijerinckii L9 do not show as many extracellular enzyme activities as Bacillus, but it performs the highest hydrogen-producing ability. The original microflora can be updated to a syntrophic bacterial co-culture system contended only with B. thermoamylovorans I and C. beijerinckii L9. The combination of aerobic Bacillus and anaerobic Clostridium may play the key role for developing the industrialized bio-fuels and bio-hydrogen-producing system from biomass.

Ten and 100 L bioreactors were utilized to evaluate the effect of stirring and pH on anaerobes converting spent brewery grains to hydrogen. Experimental results of 10-L bioreactor demonstrated that both stirring speed and pH significantly affected the hydrogen conversion and the chemical oxygen demand reduction of the spent grains. Highly reproducible metabolic data confirmed that the anaerobes from grass compost could convert the spent grains to fatty acids and alcohols as well as hydrogen. Multiple analyses indicated that stirring mainly affected the hydrogen production rate. pH level was a more important factor than the stirring on shortening hydrogen production lag and increasing hydrogen yield. Optimum conditions of pH = 6.0 and stirring speed=120 rpm were obtained for 10-L bioreactor converting 3% grains to hydrogen by observing its production rate together with yield, found to be 161 mL H 2 / gTVS / d and 13 mL H 2 / g TVS , respectively. However, the rate and the yield decreased along with stirring speeds increased above 120 rpm. When the stirring speeds of 10-L bioreactor were greater than this value, its mixing condition changed from laminar flow to turbulent flow. The 100-L experiment with a sequencing batch mode operation confirmed that the hydrogen production rate obtained with laminar flow was significantly more stable and reproducible than with turbulent flow.

A series of aromatic poly(ether imide)s containing 9,9′-spirobifluorene moieties in the main chain have been synthesized via the polycondensation of 2,2′-bis(4-aminophenoxy)-9,9′-spirobifluorene with a variety of aromatic dianhydrides. In the diamine monomer, the two aminophenoxyfluorene entities are orthogonally arranged and are connected through an sp3 carbon atom (the spiro center). The resulting poly(ether imide)s have a polymer backbone which is periodically twisted with an angle of 90° at each spiro center. This structural feature, which restricts the close packing of the polymer chains and reduces inter-chain interactions, leads to amorphous poly(ether imide)s with good solubility in common organic solvents. In addition, the rigidity of the main chain of these polymers appears to be preserved due to the spiro-structure. As a result, these poly(ether imide)s exhibit a high Tg and excellent thermal stability.

Aim: We previously characterized the molecular linkage that directs both BCL10 overexpression and nuclear translocation in response to inflammation-related NF-kB signaling pathway in breast cancer cell line, lymphoma cell line, and cervical cancer line (J Biol Chem 2006;281:167-75, Blood 2008;112:2927-34; Gynecol Oncol 2012;126:245-51). Since NF-kB activation has been shown to occur in the pathogenesis of ovarian cancer, we sought to investigate whether BCL10 possesses clinical significance in relation to ovarian cancer. Methods: Two ovarian cancer cell lines (SK-OV3, and CAOV-3) with overexpressing BCL10 were used in this study. The proliferation effect of BCL10 and its mechanism were evaluated in vitro and in vivo. The DNA binding activity of NF-kB was determined by the luciferase assay. The expression of BCL10, NF-kB, and cyclin D1 in tumor cells from an array of 100 tissue samples was examined using immunohistochemical staining. Results: We found that inhibition of BCL10 with BCL10 small interfering RNA (siRNA) significantly reduced the cell growth rate of these two ovarian cancer cell lines through the G1 arrest. BCL10 siRNA treatment inhibited the expression of p-IKKb and p-IkB, and also down-regulated NF-kB activation and its downstream cell cycle protein, cyclin D1. Furthermore, we used BCL10 short hairpin RNA (shRNA) plasmids to transfect two ovarian cancer cell lines, and found that the down-regulation of BCL10 inhibited the tumor growth rate of ovarian cancer and prolonged the survival rate in xenograft model of immunodeficient mice (SK-OV3: 38.7 ± 4.4 in WT; 59.2 ± 6.2 in BCL10 knockdown, and CAOV-3: 37.2 ± 7.8 in WT; 76.7 ± 11.6 in BCL10 knockdown). In tissue samples, we detected that serous cystadenocarcinoma had a higher rate of BCL10 nuclear expression (35/49 [71.4%]) than mucinous cystadenocarcinoma (4/18 [22.2%]) and endometrioid adenocarcinoma (16/28 [57.1%]) (P = 0.001). BCL10 nuclear expression was also closely associated with tumor grading (P = 0.017). Furthermore, nuclear expression of BCL10 was closely associated with nuclear NF-kB expression (P Conclusions: Our findings indicate that nuclear BCL10 plays an important role in controlling the growth and progression of ovarian cancer cells through NF-kB dependent cyclin D1 regulation. Disclosure: All authors have declared no conflicts of interest.

Y. Chen, S. L. Pan, J. C. Wang, C. H. Chou, S. H. Kuo, J. C. H. Cheng, C. M. Teng Department of Oncology, National Taiwan University Hospital, Taipei, Taiwan, Pharmacological Institute, College of Medicine, National Taiwan University, Taipei, Taiwan, Graduate Institute of Clinical Medicine, National Taiwan University College of Medicine, Taipei, Taiwan, Department of Internal Medicine, National Taiwan University College of Medicine, Taipei, Taiwan

Purpose/Objective: Metastasis has been the lethal factor in cancer patients. YC-1, 3-(5 -hydroxymethyl-2 -furyl)-1-benzylindazole, an agent developed for circulatory disorders by inhibiting platelet aggregation and vascular contraction, was found to inhibit hypoxia inducible factor-1 (HIF-1) activity. HIF-1 is a stress protein during radiation and modulates genes associated with cancer invasion and metastasis. With HIF-1 as a molecular target of YC-1, we try to clarify the therapeutic effect and mechanism of YC-1 on radiation induced cancer metastasis.