The Involvement of Lumican in Human Ovulatory Processes.

Based on a previous global transcriptome sequencing project, we hypothesized that Lumican (LUM) might play a role in ovulatory processes. We sought to determine LUM gene expression under various conditions in human preovulatory follicles. The in vitro expression of LUM mRNA in mural (MGCs) and cumulus (CGCs) granulosa cells was characterized using quantitative real-time polymerase chain reaction (qRT-PCR). Immunohistochemical staining was used to identify human LUM expression in follicles at different developmental stages. Cell signaling studies were performed by treating human MGCs with human chorionic gonadotropin (hCG) and both, different stimulators and inhibitors to determine their effect on LUM expression by using qRT-PCR. Cell confluence studies were carried out to study the correlation between LUM expression and follicle cell proliferation. Follicular MGCs and CGCs of women undergoing in vitro fertilization (IVF) procedures due to endometriosis were analyzed for differences in LUM expression patterns by qRT-PCR. LUM mRNA expression was significantly higher in MGCs as compared to CGCs. In CGCs, LUM mRNA was higher in mature metaphase II (MII) oocytes than in germinal vesicle (GV) and metaphase I (MI) oocytes. LUM expression was significantly upregulated in response to hCG in cultured MGCs. Immunohistochemistry of human ovaries revealed LUM was mostly present in MGCs of large preovulatory and postovulatory follicles and absent from primordial follicles. Using pharmacological activators and inhibitors, we demonstrated that LUM induction by luteinizing hormone (LH)/hCG is carried through the mitogen-activated protein kinase (MEK) pathway. LUM expression was induced in high-density cell cultures in a confluence-dependent manner. MGCs from follicles of subjects with endometriosis exhibited reduced mRNA transcription levels compared to control subjects. Our study confirms that LUM is a newly discovered ovulatory gene. LUM might play an important role during the preovulatory period up until ovulation as well as in endometriosis infertility. A better understanding of LUM's role might provide potential new treatment paradigms for some types of female infertility.

Long-term drying of Mars by sequestration of ocean-scale volumes of water in the crust.

Geological evidence shows that ancient Mars had large volumes of liquid water. Models of past hydrogen escape to space, calibrated with observations of the current escape rate, cannot explain the present-day deuterium-to-hydrogen isotope ratio (D/H). We simulated volcanic degassing, atmospheric escape, and crustal hydration on Mars, incorporating observational constraints from spacecraft, rovers, and meteorites. We found that ancient water volumes equivalent to a 100 to 1500 meter global layer are simultaneously compatible with the geological evidence, loss rate estimates, and D/H measurements. In our model, the volume of water participating in the hydrological cycle decreased by 40 to 95% over the Noachian period (~3.7 billion to 4.1 billion years ago), reaching present-day values by ~3.0 billion years ago. Between 30 and 99% of martian water was sequestered through crustal hydration, demonstrating that irreversible chemical weathering can increase the aridity of terrestrial planets.

Mars Methane Sources in Northwestern Gale Crater Inferred From Back Trajectory Modeling.

During its first seven years of operation, the Sample Analysis at Mars Tunable Laser Spectrometer (TLS) on board the Curiosity rover has detected seven methane spikes above a low background abundance in Gale crater. The methane spikes are likely sourced by surface emission within or around Gale crater. Here, we use inverse Lagrangian modeling techniques to identify upstream emission regions on the Martian surface for these methane spikes at an unprecedented spatial resolution. Inside Gale crater, the northwestern crater floor casts the strongest influence on the detections. Outside Gale crater, the upstream regions common to all the methane spikes extend toward the north. The contrasting results from two consecutive TLS methane measurements performed on the same sol point to an active emission site to the west or the southwest of the Curiosity rover on the northwestern crater floor. The observed spike magnitude and frequency also favor emission sites on the northwestern crater floor, unless there are fast methane removal mechanisms at work, or either the methane spikes of TLS or the non-detections of ExoMars Trace Gas Orbiter cannot be trusted.

Elucidating Decorin's role in the preovulatory follicle.

BACKGROUND: DCN (decorin) is a proteoglycan known to be involved in regulating cell proliferation, collagen fibril organization and migration. In our global transcriptome RNA-sequencing approach to systematically identify new ovulation-associated genes, DCN was identified as one of the highly regulated genes. We therefore hypothesize that DCN may have a role in ovulatory processes such as cell migration and proliferation. AIM: To characterize the expression, regulation and function of the proteoglycan DCN in the human ovarian follicles during the preovulatory period. METHODS: The in-vivo expression of DCN mRNA in mural (MGCs) and cumulus (CGCs) granulosa cells was characterized using quantitative RT-PCR and western blot. A signaling study was performed by treating human MGCs cultures with gonadotropins and different stimulators and inhibitors to determine their effect on DCN expression by qRT- PCR and elucidate the pathways regulating these proteins. In a functional study, KGN granulosa cell line was used to study cell migration with a scratch assay. RESULTS: DCN mRNA expression was significantly higher in MGCs compared to CGCs. DCN mRNA was significantly higher in CGCs surrounding mature metaphase II (MII) oocytes compared to CGCs of germinal vesicle (GV) and metaphase I (MI) oocytes. hCG significantly increased DCN mRNA and protein expression levels in cultured MGCs. Using signal transduction activators and inhibitors, we demonstrated that DCN induction by LH/hCG is carried out via PKA, PKC, ERK/MEK, and PI3K pathways. We showed that DCN expression is also induced in high-density cell cultures, in a dose-dependent pattern. In addition, progesterone induced a significant increase in DCN secretion to the media. MGCs from follicles of endometriosis patients exhibited reduced (about 20% of) mRNA transcriptions levels compared to MGCs follicles of control patients. More significantly, we found that DCN has an inhibiting effect on KGN cell migration. CONCLUSIONS: Our study indicates that DCN is a unique ovulatory gene. Our findings support the hypothesis that DCN plays an important new role during the preovulatory period and ovulation, and stress its involvement in endometriosis infertility. A better understanding of DCN role in ovulation and endometriosis may provide treatment for some types of infertility.

Boolean logic by convective obstacle flows.

We present a potential new mode of natural computing in which simple, heat-driven fluid flows perform Boolean logic operations. The system comprises a two-dimensional single-phase fluid that is heated from below and cooled from above, with two obstacles placed on the horizontal mid-plane. The obstacles remove all vertical momentum that flows into them. The horizontal momentum extraction of the obstacles is controlled in a binary fashion, and constitutes the 2-bit input. The output of the system is a thresholded measure of the energy extracted by the obstacles. Due to the existence of multiple attractors in the phase space of this system, the input-output relationships are equivalent to those of the OR, XOR or NAND gates, depending on the threshold and obstacle separation. The ability to reproduce these logical operations suggests that convective flows might have the potential to perform more general computations, despite the fact that they do not involve electronics, chemistry or multiple fluid phases.

Convective flow in the presence of a small obstacle: Symmetry breaking, attractors, hysteresis, and information.

This work explores the stability and hysteresis effects that occur when a small sink of momentum is introduced into a heat-driven, two-dimensional convective flow. As per standard fluid mechanical intuition, the system minimizes work generation and dissipation when one component of momentum is extracted. However, when the sink absorbs all incoming momentum, the system configures itself such that one of the convection plumes aligns directly with the sink. This state is the most hydrodynamically stable, but it maximizes, rather than minimizes extracted mechanical work. Furthermore, in the case of only vertical momentum extraction, there are two attractors, with different stabilities. Numerical experiments involving slow variations of the horizontal momentum extraction show a clear history dependence. This hysteresis preserves information about the system's past states, and hence represents a primitive memory. The momentum sink can also be used to manipulate the horizontal position of the flow field, with potential applications in microfluidics and laminar convection systems. This simple system exhibits the phenomena of autocatalysis (during the initial growth of the convection plumes), negative feedback (the attractors are either fully or quasistable), memory, and elementary computation.

Characterization of the miRNA regulators of the human ovulatory cascade.

Ovarian follicular development and ovulation are complex and tightly regulated processes that involve regulation by microRNAs (miRNAs). We previously identified differentially expressed mRNAs between human cumulus granulosa cells (CGCs) from immature early antral follicles (germinal vesicle - GV) and mature preovulatory follicles (metaphase II - M2). In this study, we performed an integrated analysis of the transcriptome and miRNome in CGCs obtained from the GV cumulus-oocyte complex (COC) obtained from IVM and M2 COC obtained from IVF. A total of 43 differentially expressed miRNAs were identified. Using Ingenuity IPA analysis, we identified 7288 potential miRNA-regulated target genes. Two hundred thirty-four of these target genes were also found in our previously generated ovulatory gene library while exhibiting anti-correlated expression to the identified miRNAs. IPA pathway analysis suggested that miR-21 and FOXM1 cooperatively inhibit CDC25A, TOP2A and PRC1. We identified a mechanism for the temporary inhibition of VEGF during ovulation by TGFB1, miR-16-5p and miR-34a-5p. The linkage bioinformatics analysis between the libraries of the coding genes from our preliminary study with the newly generated library of regulatory miRNAs provides us a comprehensive, integrated overview of the miRNA-mRNA co-regulatory networks that may play a key role in controlling post-transcriptomic regulation of the ovulatory process.

Prior disclosure and emotional expression: Interactive effects on post-traumatic stress disorder symptoms.

Background Previous research has revealed mixed findings with regard to the effects of disclosure on trauma recovery. More recently, studies on psychological trauma have found associations among religion, meaning, and health. This study investigated prior disclosure as a moderator for the association between religious emotional expression and adaptive trauma processing, as measured by post-traumatic stress disorder (PTSD) symptoms. Methods Using Pennebaker's written emotional expression paradigm, 105 participants were assigned to either a conventional trauma-writing condition or religious trauma-writing condition. PTSD symptoms were assessed at baseline and again at one-month post writing. Results A two-way interaction was found between prior disclosure and writing condition on PTSD symptoms at follow-up. For the religious trauma-writing condition only, there was a significant difference between low versus high disclosure participants in PTSD symptoms at follow-up, such that low prior disclosure participants registered fewer PTSD symptoms than high prior disclosure participants, while prior disclosure did not have such effect in the conventional trauma-writing condition. LIMITATIONS: This two-way interaction may be further qualified by other important psychosocial variables, such as differences in personality, coping style, social support, or use of prayer as a form of disclosure, which were not assessed in this study. Conclusion Religious emotional expression may encourage adaptive trauma processing, especially for individuals with low prior disclosure. These findings encourage further investigation of the conditions under which disclosure and religion may be a beneficial factor in trauma adaptation and treatment.

Determinants of aggressive behavior: Interactive effects of emotional regulation and inhibitory control.

Aggressive behavior can be defined as any behavior intended to hurt another person, and it is associated with many individual and social factors. This study examined the relationship between emotional regulation and inhibitory control in predicting aggressive behavior. Seventy-eight participants (40 males) completed self-report measures (Negative Mood Regulation Scale and Buss-Perry Aggression Questionnaire), a stop signal task, and engaged in a modified version of Taylor Aggression Paradigm (TAP) exercise, in which the outcome was used as a measure of direct physical aggression. We used a hierarchical, mixed-model multiple regression analysis test to examine the effects of emotion regulation and inhibitory control on physical reactive aggression. Results indicated an interaction between emotion regulation and inhibitory control on aggression. For participants with low inhibitory control only, there was a significant difference between high and low emotion regulation on aggression, such that low emotion regulation participants registered higher aggression than high emotion regulation participants. This difference was not found among participants with high inhibitory control. These results have implications for refining and targeting training and rehabilitation programs aimed at reducing aggressive behavior.

Assessment of psychiatric disorders among sex offenders: Prevalence and associations with criminal history.

BACKGROUND: Much of the research on the prevalence of psychiatric disorders among sex offenders has been conducted in the USA or Europe. Less is known about it in other regions, particularly in Asia. AIMS: The objective of this study is to determine the prevalence of psychiatric disorders among sex offenders in Taiwan and their associations with offender characteristics and criminal history. METHODS: Participants were randomly selected from men serving sentences in Taiwan's prison for serious sex offenders. Consenting men were assessed using the structured clinical interviews for DSM-IV-TR Axis I and II disorders. Demographics and criminal history were also recorded. RESULTS: Over two-thirds of the 68 participants met criteria for one or more lifetime Axis I disorders, and nearly 60% met criteria for one or more Axis II disorder. The higher the number of Axis I and cluster B personality disorders, the higher was the total number of convictions. CONCLUSIONS: Our study adds to the literature that suggests that psychiatric assessment is likely to have an important role in the management and treatment of sex offenders. The finding that multiple disorders are common in this group and associated with more convictions for sex offences suggests that failure to include psychiatric assessment in planning the management of sex offenders may increase the risk of recidivism.

Emotional intelligence, trauma severity, and emotional expression.

This study investigated Emotional Intelligence (EI) as a moderator for the association between emotional expression and adaptive trauma processing, as measured by depressive symptoms. Using Pennebaker's written emotional expression paradigm, 105 participants were assigned to either a conventional trauma-writing or religious trauma-writing condition. Depressive symptoms were assessed at baseline and again at one-month post writing. No significant association between EI and religiousness was found at baseline. Results indicated a three-way interaction among EI, trauma severity, and writing condition on depressive symptoms at follow-up. For the religious trauma-writing condition only, there was a significant difference between high- versus low-EI participants who experienced more severe trauma in depressive symptoms at follow-up, such that low-EI participants registered less depressive symptoms than high-EI participants; while there was no significant difference between low versus high EI for participants with less severe trauma. These findings encourage further investigation of the conditions under which religion may be a beneficial factor in trauma adaptation.

Localization of luteinizing hormone receptor protein in the human ovary.

The luteinizing hormone receptor (LHR) plays a pivotal role during follicular development. Consequently, its expression pattern is of major importance for research and has clinical implications. Despite the accumulated information regarding LHR expression patterns, our understanding of its expression in the human ovary, specifically at the protein level, is incomplete. Therefore, our aim was to determine the LHR protein localization and expression pattern in the human ovary. We examined the presence of LHR by immunohistochemical staining of human ovaries and western blots of mural granulosa and cumulus cells aspirated during IVF treatments. We were not able to detect LHR protein staining in primordial or primary follicles. We observed equivocal positive staining in granulosa cells and theca cells of secondary follicles. The first appearance of a clear signal of LHR protein was observed in granulosa cells and theca cells of small antral follicles, and there was evidence of increasing LHR production as the follicles mature to the pre-ovulatory stage. After ovulation, LHR protein was ubiquitously produced in the corpus luteum. To confirm the expression pattern in granulosa cells and cumulus cells, we performed western blots and found that LHR expression was stronger in granulosa cells than in cumulus cells, with the later demonstrating low, but still significant, amounts of LHR protein. In summary, we conclude that LHR protein starts to appear on granulosa cells and theca cells of early antral follicles, and low but significant expression of LHR exists also in the cumulus cells. These results may have implications for the future design of clinical protocols and culture mediums for in vitro fertilization and especially in vitro maturation of oocytes.

Characterization of the human cumulus cell transcriptome during final follicular maturation and ovulation.

Cumulus expansion and oocyte maturation are central processes in ovulation. Knowledge gained from rodent and other mammalian models has revealed some of the molecular pathways associated with these processes. However, the equivalent pathways in humans have not been thoroughly studied and remain unidentified. Compact cumulus cells (CCs) from germinal vesicle cumulus oocyte complexes (COCs) were obtained from patients undergoing in vitro maturation (IVM) procedures. Expanded CCs from metaphase 2 COC were obtained from patients undergoing IVF/ICSI. Global transcriptome profiles of the samples were obtained using state-of-the-art RNA sequencing techniques. We identified 1746 differentially expressed (DE) genes between compact and expanded CCs. Most of these genes were involved in cellular growth and proliferation, cellular movement, cell cycle, cell-to-cell signaling and interaction, extracellular matrix and steroidogenesis. Out of the DE genes, we found 89 long noncoding RNAs, of which 12 are encoded within introns of genes known to be involved in granulosa cell processes. This suggests that unique noncoding RNA transcripts may contribute to the regulation of cumulus expansion and oocyte maturation. Using global transcriptome sequencing, we were able to generate a library of genes regulated during cumulus expansion and oocyte maturation processes. Analysis of these genes allowed us to identify important new genes and noncoding RNAs potentially involved in COC maturation and cumulus expansion. These results may increase our understanding of the process of oocyte maturation and could ultimately improve the efficacy of IVM treatment.

Establishment and validation of a model for non-luteinized human mural granulosa cell culture.

Cell culture techniques of human mural granulosa cells (MGCs) serve as a major in vitro tool. However, the use of luteinized MGCs has major limitations due to their luteinized state. Our aim was to establish a standardized protocol for the culture of MGCs as a model for different stages of folliculogenesis. We showed that early-non-luteinized, preovulatory-non-luteinized and luteal-MGCs have distinct gene expression pattern. After 4 days of incubation of luteinized-MGCs, ovulatory genes mRNA's achieve expression levels similar to the early non-luteinized follicles. FSH stimulation for 48 h of these 4 days cultured MGCs showed ovulatory genes mRNA's expression similar to the pre-ovulatory non-luteinized follicles. These FSH-stimulated cells responded to hCG stimulation in a pattern similar to the response of pre-ovulatory follicles. This novel model may provide a standardized research tool for delineation of the molecular processes occurring during the latter stages of follicular development in the human ovary.

Exposure of fallopian tube epithelium to follicular fluid mimics carcinogenic changes in precursor lesions of serous papillary carcinoma.

OBJECTIVES: Ovulation-related inflammation is suspected to have a causal role in ovarian carcinogenesis, but there are no human models to study the molecular pathways. Our aim is to develop such an ex-vivo model based on human fallopian tube (FT) epithelium exposed to human follicular fluid (FF). METHODS: FT epithelium was dissociated from normal surgical specimens. FF was obtained from donors undergoing in-vitro fertilization. The cells were cultured on collagen-coated Transwells and incubated with FF for various periods of time. The transcriptomic changes resulting from FF treatment were profiled using Affymetrix expression arrays. Specific characteristics of the FT pre-cancerous lesions were studied using immunohistochemistry, immunofluorescence, RT-PCR and XTT assay. RESULTS: We show that FF exposure causes up-regulation of inflammatory and DNA repair pathways. Double stranded DNA breaks are induced. There is a minor increase in cell proliferation. TP53, which is the hallmark of the precursor lesion in-vivo, is accumulated. Levels of expression and secretion of Interleukin-8 are significantly increased. CONCLUSIONS: Our model addresses the main non-genetic risk factor for ovarian cancer, namely the impact of ovulation. This study demonstrates the biological implications of in-vitro exposure of human FT epithelial cells to FF. The model replicates elements characterizing the precursor lesions of ovarian cancer, and warrants further investigation of the linkage between repeated exposure to ovulation-related damage and accumulation of neoplastic changes.

Changes in the nutrient ratios and phytoplankton community after declines in nutrient concentrations in a semi-enclosed bay in Hong Kong.

Tolo Harbour is a landlocked bay with poor tidal flushings in the northeastern part of Hong Kong. During the 1980s, excessive nutrient loading led to dramatic increase in nutrient concentrations, accompanied by lower N:P ratios, higher algal biomass and shifts in the phytoplankton community. We studied the effects of nutrient loading reduction measures on nutrient concentrations, nutrient ratios and phytoplankton dynamics in Tolo Harbour by comparing data collected before the full implementation of nutrient loading reduction measures (1986-1997) to those after the implementation (1998-2008). Such measures led to declines in nutrient concentrations, changes in N:P and N:Si ratios, lower chlorophyll-a concentrations and fewer algal blooms. Diatoms were the most abundant phytoplankton group in Tolo Harbour both before and after declines in nutrient concentrations. The density of dinoflagellates did not change, but substantial increase in other algal group abundance was recorded.

TP53 regulates human AlkB homologue 2 expression in glioma resistance to Photofrin-mediated photodynamic therapy.

BACKGROUND: Photodynamic therapy (PDT) is a promising adjuvant therapy in cancer treatment. However, cancers resistant to PDT, mediated through the efflux of photosensitisers by means of P-glycoprotein or ATP-binding cassette transporter proteins, have been reported. The DNA repair has also been suggested to be responsible for PDT resistance, but little is known about the repair pathways and mechanisms involved. Therefore, this study aimed to investigate the possible function of six major DNA repair mechanisms in glioma cells resistant to Photofrin-mediated PDT (Ph-PDT). METHODS: The U87 glioma cells relatively resistant to Ph-PDT were obtained by recovering the viable cells 3 h after PDT treatment. The mRNA and protein expression levels of DNA repair genes were evaluated by quantitative real-time reverse transcription-polymerase chain reaction and western blotting, respectively. Small-interfering RNA and chromatin-immunoprecipitation assays were used to further examine the relationship between AlkB, an alkylation repair homologue 2 (Escherichia coli) (ALKBH2) and Ph-PDT responsiveness, and transcription factors involved in ALKBH2 transcription. RESULTS: The ALKBH2 of DNA damage reversal was significantly increased at both mRNA and protein levels from 30 min to 48 h post-treatment with Ph-PDT. Conversely, down-regulating ALKBH2 expression enhances Ph-PDT efficiency. Furthermore, our data clearly show for the first time that tumour protein (TP53) is directly involved by binding to the promoter of ALKBH2 in mediating Ph-PDT resistance. CONCLUSION: C The DNA damage reversal mechanisms may have important functions in Ph-PDT resistance through the activation of ALKBH2 by TP53.

Chemical dynamics of triacetylene formation and implications to the synthesis of polyynes in Titan's atmosphere.

For the last four decades, the role of polyynes such as diacetylene (HCCCCH) and triacetylene (HCCCCCCH) in the chemical evolution of the atmosphere of Saturn's moon Titan has been a subject of vigorous research. These polyacetylenes are thought to serve as an UV radiation shield in planetary environments; thus, acting as prebiotic ozone, and are considered as important constituents of the visible haze layers on Titan. However, the underlying chemical processes that initiate the formation and control the growth of polyynes have been the least understood to date. Here, we present a combined experimental, theoretical, and modeling study on the synthesis of the polyyne triacetylene (HCCCCCCH) via the bimolecular gas phase reaction of the ethynyl radical (CCH) with diacetylene (HCCCCH). This elementary reaction is rapid, has no entrance barrier, and yields the triacetylene molecule via indirect scattering dynamics through complex formation in a single collision event. Photochemical models of Titan's atmosphere imply that triacetylene may serve as a building block to synthesize even more complex polyynes such as tetraacetylene (HCCCCCCCCH).

Looking Inside the Black Box of "Attendance at Services": New Measures for Exploring an Old Dimension in Religion and Health Research.

Research in religion and health has spurred new interest in measuring religiousness. Measurement efforts have focused on subjective facets of religiousness such as spirituality and beliefs, and less attention has been paid to congregate aspects, beyond the single item measuring attendance at services. We evaluate some new measures for religious experiences occurring during congregational worship services. Respondents (N=576) were religiously-diverse community-dwelling adults interviewed prior to cardiac surgery. Exploratory factor analysis of the new items with a pool of standard items yielded a readily interpretable solution, involving seven correlated but distinct factors and one index variable, with high levels of internal consistency. We describe religious affiliation and demographic differences in these measures. Attendance at religious services provides multifaceted physical, emotional, social, and spiritual experiences that may promote physical health through multiple pathways.

Framing written emotional expression from a religious perspective: effects on depressive symptoms.

OBJECTIVE: Previous research has found an inverse association between exposure to emotional trauma and well-being. More recently, studies on written emotional expression found that repeated expression of a traumatic experience is beneficial to physical health. However, possible mechanisms through which written emotional expression may work to influence health are still under investigation. Recent research on psychological stress has also found associations among religion, meaning, and health. This study evaluated the effects of taking a religious perspective while engaged in written emotional expression. METHOD: Participants included 215 college students, who were assigned randomly to one of three experimental groups. One was given conventional written emotional expression instructions for writing about a traumatic experience; another was instructed to write about the trauma from a religious/spiritual perspective; a control group wrote about a trivial topic. RESULTS: Compared with control participants, those in the trauma-religion condition experienced significantly greater reductions in depressive symptoms at 1-month follow-up. Conventional trauma writing had no effect on dependent measures. CONCLUSION: These findings encourage further investigation of religion as a factor that may augment the health-promoting effects of written emotional expression.