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author:("ohi, S")
1.  Relation between Isolated Venous Thrombi in Soleal Muscle and Positive Anti-Nuclear Antibody 
Annals of Vascular Diseases  2012;5(3):321-327.
Objective: In patients with isolated soleal vein thrombosis (SVT), the relation between acute thrombi and positive anti-nuclear antibody (ANA) was investigated.
Subjects and Methods: The subjects were 116 lower extremities in 86 patients with SVT. They were diagnosed and examined by ultrasonography and blood serum analysis (D-dimer, ANA), and had been followed up every three months.
Results: They had acute SVT in 35 limbs (30%) and chronic SVT in 86 limbs (70%), and they had positive ANA in 63%. They had recurrent SVT in 26%, and all were positive for ANA.
Conclusion: ANA-positivity might be a risk factor for acute thrombi in patients with SVT. (*English Translation of J Jpn Coll Angiol 2010; 50: 417-422.)
doi:10.3400/avd.oa.12.00052
PMCID: PMC3595847  PMID: 23555531
isolated soleal vein thrombosis; recurrent deep vein thrombosis; risk factor; antinuclear antibody
2.  LncRNA-Dependent Mechanisms of Androgen Receptor-regulated Gene Activation Programs 
Nature  2013;500(7464):598-602.
While recent studies indicated roles of long non-coding RNAs (lncRNAs) in physiologic aspects of cell-type determination and tissue homeostasis1 yet their potential involvement in regulated gene transcription programs remain rather poorly understood. Androgen receptor (AR) regulates a large repertoire of genes central to the identity and behavior of prostate cancer cells2, and functions in a ligand-independent fashion in many prostate cancers when they become hormone refractory after initial androgen deprivation therapy3. Here, we report that two lncRNAs highly overexpressed in aggressive prostate cancer, PRNCR1 and PCGEM1, bind successively to the AR and strongly enhance both ligand-dependent and ligand-independent AR-mediated gene activation programs and proliferation in prostate cancer cells. Binding of PRNCR1 to the C-terminally acetylated AR on enhancers and its association with DOT1L appear to be required for recruitment of the second lncRNA, PCGEM1, to the DOT1L-mediated methylated AR N-terminus. Unexpectedly, recognition of specific protein marks by PCGEM1-recruited Pygopus2 PHD domain proves to enhance selective looping of AR-bound enhancers to target gene promoters in these cells. In “resistant” prostate cancer cells, these overexpressed lncRNAs can interact with, and are required for, the robust activation of both truncated and full length AR, causing ligand-independent activation of the AR transcriptional program and cell proliferation. Conditionally-expressed short hairpin RNA (shRNA) targeting of these lncRNAs in castration-resistant prostate cancer (CRPC) cell lines strongly suppressed tumor xenograft growth in vivo. Together, these results suggest that these overexpressed lncRNAs can potentially serve as a required component of castration-resistance in prostatic tumors.
doi:10.1038/nature12451
PMCID: PMC4034386  PMID: 23945587
Nuclear Receptor; Non-coding RNA; Enhancer; Pygopus2; Transcription Activation; Castration-resistant Prostate Cancer
3.  Evaluation of Influence of Single Nucleotide Polymorphisms in Cytochrome P450 2B6 on Substrate Recognition Using Computational Docking and Molecular Dynamics Simulation 
PLoS ONE  2014;9(5):e96789.
In this study, we investigated the influence of single nucleotide polymorphisms on the conformation of mutated cytochrome P450 (CYP) 2B6 proteins using molecular dynamics (MD) simulation. Some of these mutations influence drug metabolism activities, leading to individual variations in drug efficacy and pharmacokinetics. Using computational docking, we predicted the structure of the complex between the antimalarial agent artemether and CYP2B6 whose conformations were obtained by MD simulation. The simulation demonstrated that the entire structure of the protein changes even when a single residue is mutated. Moreover, the structural flexibility is affected by the mutations and it may influence the enzyme activity. The results suggest that some of the inactive mutants cannot recognize artemether due to structural changes caused by the mutation.
doi:10.1371/journal.pone.0096789
PMCID: PMC4010486  PMID: 24796891
4.  Functional Importance of eRNAs for Estrogen-dependent Transcriptional Activation Events 
Nature  2013;498(7455):516-520.
The functional importance of gene enhancers in regulated gene expression is well established(1–3). In addition to widespread transcription of long non-coding RNAs (ncRNA) in mammalian cells(4–6), bidirectional ncRNAs referred to as eRNAs are transcribed on enhancers(7–9). However, it has remained unclear whether these eRNAs are functional, or merely a reflection of enhancer activation. Here, we report that 17β-estradiol (E2)-bound estrogen receptor α (ERα) on enhancers causes a global increase in eRNA transcription on enhancers adjacent to E2-upregulated coding genes. These induced eRNAs, as functional transcripts, appear to exert important roles for the observed ligand-dependent induction of target coding genes, causing an increased strength of specific enhancer:promoter looping initiated by ERα binding. Cohesin, present on many ERα-regulated enhancers even prior to ligand treatment, apparently contributes to E2-dependent gene activation, at least in part, by stabilizing E2/ERα/eRNA-induced enhancer:promoter looping. Our data indicate that eRNAs are likely to exert important functions in many regulated programs of gene transcription.
doi:10.1038/nature12210
PMCID: PMC3718886  PMID: 23728302
5.  Changes in serum BDNF levels associated with moderate-intensity exercise in healthy young Japanese men 
SpringerPlus  2013;2:678.
Purpose
The purpose of this study was to examine the influence of acute moderate-intensity exercise on serum brain-derived neurotrophic factor (BDNF) levels in healthy young Japanese men. BDNF is one of a family of neurotrophic factors involved in neuronal transmission, modulation and plasticity. Previous human-based studies have demonstrated that acute exercise leads to increases in BDNF; however, to date there has been no study conducted among Japanese male subjects.
Methods
Forty young adult Japanese men (aged 24.1 ± 2.9 years) – reduced to a total of thirty three following subjection to exclusion criteria – performed 30 minutes of exercise at 60% of VO2max on a stationary bicycle. Serum BDNF was analyzed both before and after exercise.
Results
Eighteen of the total thirty three subjects demonstrated an increase in serum BDNF after exercise. However, on aggregate, the change in serum BDNF associated with exercise was not significant (p = .17).
Conclusions
This is the first study to demonstrate that serum BDNF levels are not consistently increased by acute moderate-intensity exercise in Japanese men. It is likely that something in the lifestyle and/or environment of male Japanese subjects underlies the difference between our findings and studies conducted in other countries.
doi:10.1186/2193-1801-2-678
PMCID: PMC3872285  PMID: 24386624
Brain-derived neurotrophic factor; Japanese; Human; Physical activity; Aerobic exercise
6.  A novel platform to enable inhaled naked RNAi medicine for lung cancer 
Scientific Reports  2013;3:3325.
Small interfering RNA (siRNA)-based therapeutics have been used in humans and offer distinct advantages over traditional therapies. However, previous investigations have shown that there are several technical obstacles that need to be overcome before routine clinical applications are used. Currently, we are launching a novel class of RNAi therapeutic agents (PnkRNA™, nkRNA) that show high resistance to degradation and are less immunogenic, less cytotoxic, and capable of efficient intracellular delivery. Here, we develop a novel platform to promote naked RNAi approaches administered through inhalation without sophisticated delivery technology in mice. Furthermore, a naked and unmodified novel RNAi agent, such as ribophorin II (RPN2-PnkRNA), which has been selected as a therapeutic target for lung cancer, resulted in efficient inhibition of tumor growth without any significant toxicity. Thus, this new technology using aerosol delivery could represent a safe, potentially RNAi-based strategy for clinical applications in lung cancer treatment without delivery vehicles.
doi:10.1038/srep03325
PMCID: PMC3839038  PMID: 24270189
7.  DICER- and AGO3-dependent Generation of Retinoic Acid-Induced DR2 Alu RNAs (riRNAs) Regulates Human Stem Cell Proliferation 
Nature structural & molecular biology  2012;19(11):1168-1175.
While liganded nuclear receptors are established to regulate Pol II-dependent transcription units, their role in regulating Pol III-transcribed DNA repeats remains largely unknown. Here, we report that ~2–3% of the ~100,000–200,000 human DR2 Alu repeats in proximity to activated Pol II transcription units are activated by retinoic acid receptor in human embryonic stem cells to generate Pol III-dependent RNAs. These transcripts are processed, initially in a DICER-dependent fashion, into small RNAs (~28–65nt.), referred to as riRNAs, that cause degradation of a subset of critical stem cell mRNAs, including Nanog, modulating exit from the proliferative stem cell state. This regulation requires AGO3-dependent accumulation of processed DR2 Alu transcripts and subsequent recruitment of AGO3-associated decapping complexes to the target mRNA. In this way, the RAR and Pol III-dependent DR2 Alu transcriptional events in stem cells functionally complement the Pol II-dependent neuronal transcriptional program.
doi:10.1038/nsmb.2400
PMCID: PMC3743530  PMID: 23064648
8.  Synthesis of 18O-labeled RNA for application to kinetic studies and imaging 
Nucleic Acids Research  2013;41(12):e126.
Radioisotopes and fluorescent compounds are frequently used for RNA labeling but are unsuitable for clinical studies of RNA drugs because of the risk from radiation exposure or the nonequivalence arising from covalently attached fluorophores. Here, we report a practical phosphoramidite solid-phase synthesis of 18O-labeled RNA that avoids these disadvantages, and we demonstrate its application to quantification and imaging. The synthesis involves the introduction of a nonbridging 18O atom into the phosphate group during the oxidation step of the synthetic cycle by using 18O water as the oxygen donor. The 18O label in the RNA was stable at pH 3–8.5, while the physicochemical and biological properties of labeled and unlabeled short interfering RNA were indistinguishable by circular dichroism, melting temperature and RNA-interference activity. The 18O/16O ratio as measured by isotope ratio mass spectrometry increased linearly with the concentration of 18O-labeled RNA, and this technique was used to determine the blood concentration of 18O-labeled RNA after administration to mice. 18O-labeled RNA transfected into human A549 cells was visualized by isotope microscopy. The RNA was observed in foci in the cytoplasm around the nucleus, presumably corresponding to endosomes. These methodologies may be useful for kinetic and cellular-localization studies of RNA in basic and pharmaceutical studies.
doi:10.1093/nar/gkt344
PMCID: PMC3695515  PMID: 23632164
9.  Differences in dual-task performance and prefrontal cortex activation between younger and older adults 
BMC Neuroscience  2013;14:10.
Background
The purpose of this study was to examine task-related changes in prefrontal cortex (PFC) activity during a dual-task in both healthy young and older adults and compare patterns of activation between the age groups. We also sought to determine whether brain activation during a dual-task relates to executive/attentional function and how measured factors associated with both of these functions vary between older and younger adults.
Results
Thirty-five healthy volunteers (20 young and 15 elderly) participated in this study. Near-infrared spectroscopy (NIRS) was employed to measure PFC activation during a single-task (performing calculations or stepping) and dual-task (performing both single-tasks at once). Cognitive function was assessed in the older patients with the Trail-making test part B (TMT-B). Major outcomes were task performance, brain activation during task (oxygenated haemoglobin: Oxy-Hb) measured by NIRS, and TMT-B score. Mixed ANOVAs were used to compare task factors and age groups in task performance. Mixed ANOVAs also compared task factors, age group and time factors in task-induced changes in measured Oxy-Hb. Among the older participants, correlations between the TMT-B score and Oxy-Hb values measured in each single-task and in the dual-task were examined using a Pearson correlation coefficient.
Oxy-Hb values were significantly increased in both the calculation task and the dual-task within patients in both age groups. However, the Oxy-Hb values associated with there were higher in the older group during the post-task period for the dual-task. Also, there were significant negative correlations between both task-performance accuracy and Oxy-Hb values during the dual-task and participant TMT-B scores.
Conclusions
Older adults demonstrated age-specific PFC activation in response to dual-task challenge. There was also a significant negative correlation between PFC activation during dual-task and executive/attentional function. These findings suggest that the high cognitive load induced by dual-task activity generates increased PFC activity in older adults. However, this relationship appeared to be strongest in participants with better baseline attention and executive functions.
doi:10.1186/1471-2202-14-10
PMCID: PMC3552708  PMID: 23327197
Dual-task; Near-infrared spectroscopy; Executive function; Attentional function
10.  A protective strategy against hyperinflammatory responses requiring the non-transcriptional actions of GPS2 
Molecular cell  2012;46(1):91-104.
SUMMARY
The association between hyper-inflammatory states and numerous diseases is widely recognized, but our understanding of the molecular strategies that have evolved to prevent uncontrolled activation of inflammatory responses remains incomplete. Here, we report a critical, non-transcriptional role of GPS2 as a guardian against hyperstimulation of the TNFα-induced gene program. GPS2 cytoplasmic actions are required to specifically modulate RIP1 ubiquitylation and JNK activation by inhibiting TRAF2/Ubc13 enzymatic activity. In vivo relevance of GPS2 anti-inflammatory role is confirmed by inhibition of TNFα target genes in macrophages and by improved insulin signaling in the adipose tissue of aP2-GPS2 transgenic mice. As the non-transcriptional role is complemented by GPS2 functioning as positive and negative cofactor for nuclear receptors, in vivo overexpression also results in elevated circulating level of Resistin and development of hepatic steatosis. Together, these studies define GPS2 as a molecular guardian required for precise control of inflammatory responses involved in immunity and homeostasis.
doi:10.1016/j.molcel.2012.01.025
PMCID: PMC3327812  PMID: 22424771
11.  Efficacy of a Novel Class of RNA Interference Therapeutic Agents 
PLoS ONE  2012;7(8):e42655.
RNA interference (RNAi) is being widely used in functional gene research and is an important tool for drug discovery. However, canonical double-stranded short interfering RNAs are unstable and induce undesirable adverse effects, and thus there is no currently RNAi-based therapy in the clinic. We have developed a novel class of RNAi agents, and evaluated their effectiveness in vitro and in mouse models of acute lung injury (ALI) and pulmonary fibrosis. The novel class of RNAi agents (nkRNA®, PnkRNA™) were synthesized on solid phase as single-stranded RNAs that, following synthesis, self-anneal into a unique helical structure containing a central stem and two loops. They are resistant to degradation and suppress their target genes. nkRNA and PnkRNA directed against TGF-β1mRNA ameliorate outcomes and induce no off-target effects in three animal models of lung disease. The results of this study support the pathological relevance of TGF-β1 in lung diseases, and suggest the potential usefulness of these novel RNAi agents for therapeutic application.
doi:10.1371/journal.pone.0042655
PMCID: PMC3419724  PMID: 22916145
12.  ncRNA- and Pc2 Methylation-Dependent Gene Relocation between Nuclear Structures Mediates Gene Activation Programs 
Cell  2011;147(4):773-788.
Summary
Although eukaryotic nuclei contain distinct architectural structures associated with noncoding RNAs (ncRNAs), their potential relationship to regulated transcriptional programs remains poorly understood. Here, we report that methylation/demethylation of Polycomb 2 protein (Pc2) controls relocation of growth control genes between Polycomb bodies (PcGs) and interchromatin granules (ICGs) in response to growth signals. This movement is the consequence of binding of methylated and unmethylated Pc2 to the ncRNAs, TUG1 and MALAT1/NEAT2, located in PcGs and ICGs, respectively. These ncRNAs mediate assembly of multiple co-repressors/co-activators, and can alter the histone marks read by Pc2 in vitro. Additionally, binding of NEAT2 to unmethylated Pc2 promotes E2F1 SUMOylation, leading to activation of the growth control gene program. These observations delineate a molecular pathway linking the actions of subnuclear structure-specific ncRNAs and non-histone protein methylation to relocation of transcription units in the three-dimensional space of the nucleus, thus achieving coordinated gene expression programs.
doi:10.1016/j.cell.2011.08.054
PMCID: PMC3297197  PMID: 22078878
Noncoding RNA; Nuclear Architecture; Subnuclear Structures; Suv39h1; Pc2; Methylation; KDM4C; Polycomb Body; Interchromatin Granules; E2F1; SUMOylation; Ubiquitination
13.  Reprogramming Transcription via Distinct Classes of Enhancers Functionally Defined by eRNA 
Nature  2011;474(7351):390-394.
Mammalian genomes are populated with thousands of transcriptional enhancers that orchestrate cell type-specific gene expression programs1-4, but how those enhancers are exploited to institute alternative, signal-dependent transcriptional responses remains poorly understood. Here we present evidence that cell lineage-specific factors, such as FoxA1, can simultaneously facilitate and restrict key regulated transcription factors, exemplified by the androgen receptor (AR), to act on structurally- and functionally-distinct classes of enhancers. Consequently, FoxA1 down-regulation, an unfavorable prognostic sign in certain advanced prostate tumors, triggers dramatic reprogramming of the hormonal response by causing a massive switch in AR binding to a distinct cohort of pre-established enhancers. These enhancers are functional, as evidenced by the production of enhancer-templated non-coding RNA (eRNA5) based on global nuclear-on (GRO-seq) analysis6, with a unique class apparently requiring no nucleosome remodeling to induce specific enhancer-promoter looping and gene activation. GRO-seq data also suggest that liganded AR induces both transcription initiation and elongation. Together, these findings reveal a large repository of active enhancers that can be dynamically tuned to elicit alternative gene expression programs, which may underlie many sequential gene expression events in development, cell differentiation and disease progression.
doi:10.1038/nature10006
PMCID: PMC3117022  PMID: 21572438
14.  PHF8 Mediates Histone H4 Lysine 20 Demethylation Events Involved in Cell Cycle Progression 
Nature  2010;466(7305):508-512.
While reversible histone modifications are linked to an ever-expanding range of biological functions1–5, the demethylases for histone H4 lysine 20 and their potential regulatory roles remain unknown. Here, we report that the PHD and Jumonji C (JmjC) domain-containing protein, PHF8, while utilizing multiple substrates, including H3K9me1/2 and H3K27me2, also functions as an H4K20me1 demethylase. PHF8 is recruited to promoters by its PHD domain based on interaction with H3K4me2/3 and controls G1/S transition in conjunction with E2F1, HCF-1 and Set1A, at least in part, by removing the repressive H4K20me1 mark from a subset of E2F1-regulated gene promoters. Phosphorylation-dependent PHF8 dismissal from chromatin in prophase is apparently required for the accumulation of H4K20me1 during early mitosis, which might represent a component of the Condensin II loading process. Accordingly, the HEAT repeat clusters in two non-SMC Condensin II subunits, N-CAPD3 and N-CAPG2, are capable of recognizing H4K20me1, and ChIP-seq. analysis demonstrate a significant overlap of Condensin II and H4K20me1 sites in mitotic HeLa cells. Thus, the identification and characterization of the first H4K20me1 demethylase, PHF8, has revealed an intimate link between this enzyme and two distinct events in cell cycle progression.
doi:10.1038/nature09272
PMCID: PMC3059551  PMID: 20622854
15.  Synthesis and biological activity of artificial mRNA prepared with novel phosphorylating reagents 
Nucleic Acids Research  2010;38(21):7845-7857.
Though medicines that target mRNA are under active investigation, there has been little or no effort to develop mRNA itself as a medicine. Here, we report the synthesis of a 130-nt mRNA sequence encoding a 33-amino-acid peptide that includes the sequence of glucagon-like peptide-1, a peptide that stimulates glucose-dependent insulin secretion from the pancreas. The synthesis method used, which had previously been developed in our laboratory, was based on the use of 2-cyanoethoxymethyl as the 2′-hydroxy protecting group. We also developed novel, highly reactive phosphotriester pyrophosphorylating reagents to pyrophosphorylate the 5′-end of the 130-mer RNA in preparation for capping. We completed the synthesis of the artificial mRNA by the enzymatic addition of a 5′-cap and a 3′-poly(A) tail to the pyrophosphorylated 130-mer and showed that the resulting mRNA supported protein synthesis in a cell-free system and in whole cells. As far as we know, this is the first time that mRNA has been prepared from a chemically synthesized RNA sequence. As well as providing a research tool for the intracellular expression of peptides, the technology described here may be used for the production of mRNA for medical applications.
doi:10.1093/nar/gkq638
PMCID: PMC2995060  PMID: 20660478
16.  Nuclear Receptor-Induced Chromosomal Proximity and DNA Breaks Underlie Specific Translocations in Cancer 
Cell  2009;139(6):1069-1083.
Summary
Chromosomal translocations are a hallmark of leukemia/lymphoma and also appear in solid tumors, but the underlying mechanism remains elusive. By establishing a cellular model that mimics the relative frequency of authentic translocation events without proliferation selection, we report mechanisms of nuclear receptor-dependent tumor translocations. Intronic binding of liganded-AR first juxtaposes translocation loci by triggering intra- and interchromosomal interactions. AR then promotes site-specific DNA double-stranded breaks (DSBs) at translocation loci by recruiting two types of enzymatic machinery induced by genotoxic stress and liganded-AR, including Activation-Induced Cytidine Deaminase (AID) and the LINE-1 repeat-encoded ORF2 endonuclease. These enzymatic machineries synergistically generate site-selective DSBs at juxtaposed translocation loci that are ligated by Non-Homologous Ending Joining (NHEJ) pathway for specific translocations. Our data suggest that the confluence of two parallel pathways initiated by liganded-nuclear receptor and genotoxic stress underlie non-random tumor translocations, which may function in many types of tumors and pathological processes.
doi:10.1016/j.cell.2009.11.030
PMCID: PMC2812435  PMID: 19962179
Tumor Translocations; Prostate Cancer; Androgen Receptor; AID; PIWIL1; Nuclear Architecture; LINE-1 Repeats; ORF2; DNA Damage/Repair; LNCaP Cells
17.  The factorial structure of the mini mental state examination (MMSE) in Japanese dementia patients 
BMC Geriatrics  2010;10:36.
Background
The Mini-Mental State Examination (MMSE) is one of the most commonly used instruments in the evaluation of global cognitive status. Few studies have investigated the relationship among its components in terms of factorial structure in Japanese individuals suffering from dementia. The aims of this study were: 1) to analyze the factorial structure of MMSE in Japanese dementia patients, 2) to clarify the MMSE static structure in identifying different cognitive profiles and understanding how these profiles are related to levels of dysfunction in subsets of dementia patients.
Methods
30,895 consecutive outpatients with dementia were evaluated. The 11 subtests composing the MMSE and the global MMSE score were analyzed. Factor analysis based on principal component analysis with Promax rotation was applied to the data representing the frequency of failures in each subtest as identified by the MMSE.
Results
Factor analysis identified three factors that explained approximately 44.57% of the total variance. The first factor, immediate memory, essentially constituted a simple index of the reading and writing subtests. The second factor, orientation and delayed recall, expressed the ability to handle new information. The third factor, working memory, was most closely related to the severity of dementia at the time of test administration.
Conclusions
Japanese dementia patients appear to develop difficulty handling new information in the early stages of their disease. This finding, and our finding that there is a factor associated with disease severity, suggest that understanding the specific factors related to subtest items, which underlie the total MMSE score may be useful to clinicians in planning interventions for Japanese patients in the early stages of dementia.
doi:10.1186/1471-2318-10-36
PMCID: PMC2903593  PMID: 20534132
18.  Frontal brain activation in young children during picture book reading with their mothers 
Aim
This study was to measure changes in frontal brain activation in young children during picture book reading with their mothers.
Methods
The cross-sectional sample consisted of 15 young Japanese children (8 girls and 7 boys, mean age 23.1±3.4). Two experimental tasks were presented as follows: Task 1 (picture book reading with their mothers); Task 2 (viewing of book-on-video). Duration of task stimulus was 180 sec and the 60 sec interval was filled. Brain activation was measured using an optical topography system.
Results
Significant increases in oxy-Hb were observed in both right and left frontal areas in response to Task 1 compared to Task 2. There were significant correlations between child’s brain activity and mothers’ and children’s verbal – nonverbal behaviors.
Conclusion
There was greater frontal lobe activation in children when they were engaged in a picture book reading task with their mothers, as opposed to passive viewing of a videotape in which the story was read to them. Social and verbal engagement of the mother in reading picture books with her young child may mediate frontal brain activity in the child.
doi:10.1111/j.1651-2227.2009.01562.x
PMCID: PMC2848291  PMID: 19849672
picture book reading; verbal – nonverbal behaviors; frontal brain activation; near-infrared spectroscopy (NIRS)
19.  The ratio of late-follicular to mid-follicular phase LH concentrations efficiently predicts ART outcomes in women undergoing ART treatment with GnRH-agonist long protocol and stimulation with recombinant FSH 
Purpose
To establish an index to predict ART outcomes and to identify infertile patients who need LH supplementation during ovarian stimulation.
Methods
Serum LH concentrations were measured during the mid- and late-follicular phase in 86 normogonadotropic infertile patients who underwent ART treatment using GnRH-agonist long protocol with recombinant-FSH. The relationships between serum LH concentrations at both time points and ART outcomes were retrospectively analyzed, and the relationships between the ratio of late-follicular to mid-follicular LH concentrations and ART outcomes were also evaluated.
Results
There were no significant correlations between the mid- or late-follicular LH concentrations and ART outcomes. The ratio of late-follicular to mid-follicular LH concentrations <1.0 was considered the relatively LH decreased group (RD group) and ratio ≥ 1.0 was considered the relatively LH increased group (RI group). The number of usable embryos in RD group was similar to that in RI group, but the pregnancy and implantation rates in the RD group (9.7% and 5.8%) were significantly lower than those in the RI group (31.1% and 17.2%; p < 0.05).
Conclusions
Relatively decreased LH concentrations during ovarian stimulation using GnRH-agonist long protocol with rec-FSH had a negative effect on ART outcomes. Therefore, the ratio of mid- to late-follicular phase LH concentrations is suggested to be an efficient index to identify patients who might benefit from LH supplementation.
doi:10.1007/s10815-008-9243-0
PMCID: PMC2582123  PMID: 18752065
ART outcome; GnRH-agonist; LH concentration; Long protocol; Recombinant-FSH; Pregnancy rate
20.  A Histone H2A Deubiquitinase Complex Coordinating Histone Acetylation and H1 Dissociation in Transcriptional Regulation 
Molecular cell  2007;27(4):609-621.
Summary
Deciphering the epigenetic “code” remains a central issue in transcriptional regulation. Here, we report the identification of a MPN+/JAMM domain-containing histone H2A deubiquitinase (2A-DUB, or KIAA1915/MYSM1) specific for monoubiquitinated H2A (uH2A), that has permitted delineation of a strategy for specific regulatory pathways of gene activation. 2A-DUB regulates transcription by coordinating histone acetylation and deubiquitination, and destabilizing the association of linker histone H1 with nucleosomes. 2A-DUB interacts with p/CAF in a co-regulatory protein complex, with its deubiquitinase activity modulated by the status of acetylation of nucleosomal histones. Consistent with this mechanistic role, 2A-DUB participates in transcriptional regulation events in androgen receptor-dependent gene activation, and the levels of uH2A are dramatically decreased in prostate tumors, serving as a cancer-related mark. We suggest that H2A ubiquitination represents a widely-used mechanism for many regulatory transcriptional programs, and predict that various H2A ubiquitin ligases/deubiquitinases will be identified for specific cohorts of regulated transcription units.
doi:10.1016/j.molcel.2007.07.024
PMCID: PMC2709280  PMID: 17707232
21.  The frequency of ovulation from the affected ovary decreases following laparoscopic cystectomy in infertile women with unilateral endometrioma during a natural cycle 
Purpose
To evaluate the cystectomy-induced damage on the follicular growth and ovulation of an affected ovary during natural cycles.
Methods
Twenty-eight infertile patients with unilateral ovarian endometriomas who underwent laparoscopic cystectomy were retrospectively evaluated. The ovulation rate of an affected ovary during natural cycles was compared before and after cystectomy in each patient, and it was also determined if ovulation from the affected ovaries resulted in pregnancy.
Results
After surgery, the ovulation rate was significantly lower than that before cystectomy (16.9 ± 4.5% vs. 34.4 ± 6.6%, P = 0.013). After surgery, 14 pregnancies were achieved without IVF treatment, and only 2 of them (14.3%) were achieved from an operated-side ovary. However, the pregnancy rate per ovulatory cycle of the operated-side ovary was not different from that of the intact ovary (8.8% vs. 5.8%, P = 0.750).
Conclusions
Laparoscopic cystectomy is an invasive treatment in that it reduces the frequency of ovulation; however the pregnancy rate per ovulation did not deteriorate.
doi:10.1007/s10815-008-9229-y
PMCID: PMC2582085  PMID: 18563551
Endometrioma; Enucleation; Natural cycle; Ovarian reserve; Ovulation rate
22.  Elevated basal FSH levels, if it is under 15 IU/L, will not reflect poor ART outcomes 
Purpose
For this study, the impact of basal FSH levels on ART outcomes was assessed.
Methods
From June 2003 to May 2006, 191 ART cycles were performed in our hospital. All cases were treated with GnRH-a long protocol. The patients were classified according to their basal FSH level as follows: group A: FSH < 10 IU/l, group B: 10 ≦ FSH < 15 IU/l, and group C: 15 IU/l ≦ FSH. ART outcomes were compared among the three groups.
Results
The number of retrieved oocytes in group A was significantly higher than in group B, but fertilized oocytes and the pregnancy rates were comparable. The pregnancy rate in group C was not significantly lower than those found in either group A or B, but the trend was lower.
Conclusion
Oocytes retrieved from the patients who showed basal FSH levels below 15 IU/l were found to possess significant pregnancy potential.
doi:10.1007/s10815-007-9195-9
PMCID: PMC2582117  PMID: 18228128
ART; Basal FSH; Pregnancy rate
23.  TBL1 and TBLR1 Phosphorylation on Regulated Gene Promoters Overcomes Dual CtBP and NCoR/SMRT Transcriptional Repression Checkpoints 
Molecular cell  2008;29(6):755-766.
A key strategy to achieve regulated gene expression in higher eukaryotes is to prevent illegitimate signal-independent activation by imposing robust control on the dismissal of corepressors. Here, we report that many signaling pathways, including Notch, NFkB, and nuclear receptor ligands, are subjected to a dual repression “check point” based on distinct corepressor complexes. Gene activation requires the release of both CtBP1/2- and NCoR/SMRT-dependent repression, through the coordinate action of two highly related exchange factors, the transducer β-like proteins TBL1 and TBLR1, that license ubiquitylation and degradation of CtBP1/2 and NCoR/SMRT, respectively. Intriguingly, their function and differential specificity resides in only five specific Ser/Thr phosphorylation site differences, regulated by direct phosphorylation at the level of the promoter, as exemplified by the role of PKCδ in TBLR1-dependent dismissal of NCoR. Thus, our data reveal a strategy of dual- factors repression checkpoints, in which dedicated exchange factors serve as sensors for signal- specific dismissal of distinct corepressors, with specificity imposed by upstream signaling pathways.
doi:10.1016/j.molcel.2008.01.020
PMCID: PMC2364611  PMID: 18374649
TBL1; TBLR1; nuclear receptors; N-CoR; CtBP; ubiquitylation; repression; transcription
24.  Histone H2A Monoubiquitination Represses Transcription by Inhibiting RNA Polymerase II Transcriptional Elongation 
Molecular cell  2008;29(1):69-80.
SUMMARY
Solving the biological roles of covalent histone modifications, including monoubiqutination of histone H2A, and the molecular mechanisms by which these modifications regulate specific transcriptional programs, remains a central question for all eukaryotes. Here, we report that the N-CoR/HDAC1/3 complex specifically recruits a previously-unidentified specific histone H2A ubiquitin ligase, 2A-HUB/hRUL138, to a subset of regulated gene promoters. 2A-HUB catalyzes monoubiquitination of H2A at lysine 119, functioning as a combinatoric component of the repression machinery required for specific gene regulation programs. Thus, 2A-HUB mediates a selective repression of a specific set of chemokine genes in macrophages, critically modulating migratory responses to TLR activation. H2A monoubiquitination acts to prevent FACT recruitment at the transcriptional promoter region, blocking RNA polymerase II release at the early stage of elongation. We suggest that distinct H2A ubiquitinases, each recruited based on interactions with different corepressor complexes, contribute to effective distinct transcriptional repression programs.
doi:10.1016/j.molcel.2007.11.002
PMCID: PMC2327256  PMID: 18206970
25.  Reduction of perifollicular arterial blood flow resistance after hCG administration is a good indicator of the recovery of mature oocytes in ART treatment 
Puerose: To demonstrate our hypothesis that a correlation exists between oocyte maturity and a decrease in intraovarian blood flow resistance in assisted reproductive technology (ART) treatment cycles, oocyte maturity and total antioxidant status (TAOS) in follicular fluid was examined.
Methods: A total of 59 cycles involving 51 infertile patients undergoing ART treatment in our hospital were recruited in this study. The ART cycles were divided into two groups: deeply decreased (DD) group and not-deeply decreased (NDD) group, according to the pulsatility index (PI) values for perifollicular arterial blood flow before and after hCG administration. The PI values that showed a decrease in their PI after hCG administration of 10% or more were defined “deeply decreased” and showed a decrease of less than 10% were considered “not-deeply decreased.” The recovery rates of mature and immature oocytes and TAOS in follicular fluid were compared between the two groups.
Results: Mature and immature oocyte recovery rates in the DD group (62.5% and 17.2%) were significantly higher and lower, respectively, than those in the NDD group (41.7% and 38.3%, p < 0.05). The TAOS (1.55 ± 0.44 mmol/L) of the DD group was significantly lower than that in the NDD group (1.68 ± 0.47 mmol/L, p < 0.05).
Conclusions: The decrease in intraovarian arterial blood flow resistance measured after hCG administration may be a good indicator of retrieving mature oocyte.
doi:10.1007/s10815-006-9087-4
PMCID: PMC3455098  PMID: 17146737
ART; Intraovarian blood flow resistance; Oocyte maturity; Total antioxidant status (TAOS)

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