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In what traits do people interact with others who are similar to them in completely anonymous online communication? Can those traits contribute to greater exchange of opinion and information across the sociodemographic boundaries that often limit interaction between social strata? To answer this question concerning online homophily, we combined survey data on 7,287 users (aged 18 and above) of a Korean online dating advice platform with their behavioral data from June 2015 to August 2015 and explored whether advice exchange occurred between users with similar sociodemographic and personality traits. On this platform, two types of interactions occurred as follows: (1) responses to a randomly distributed problem submitted by an advice seeker and (2) the seeker's indication of approval of any of the responses given. The study found that (1) a receiver was more likely to respond to problems submitted by seekers of a comparable age and that (2) seekers were more likely to approve of a response if the seeker and receiver had similar educational backgrounds. By contrast, homophily based on personality traits was not observed even though some personality traits significantly affected the likelihood of both response and approval. Our findings suggest that online communication may breed sociodemographic homophily, whether based on age or education, more than expected or intended while not easily fostering alternative forms of homophily, such as personality homophily, which can potentially cut across borders dividing sociodemographic groups.

Transforming growth factor-beta (TGFβ) is an enigmatic protein with various roles in healthy tissue homeostasis/development as well as the development or progression of cancer, wound healing, fibrotic disorders, and immune modulation, to name a few. As TGFβ is causal to various fibroproliferative disorders featuring localized or systemic tissue/organ fibrosis as well as the activated stroma observed in various malignancies, characterizing the pathways and players mediating its action is fundamental. In the current study, we found that TGFβ induces the expression of the immunoinhibitory molecule Programed death-ligand 1 (PD-L1) in human and murine fibroblasts in a Smad2/3- and YAP/TAZ-dependent manner. Furthermore, PD-L1 knockdown decreased the TGFβ-dependent induction of extracellular matrix proteins, including collagen Iα1 (colIα1) and alpha-smooth muscle actin (α-SMA), and cell migration/wound healing. In addition to an endogenous role for PD-L1 in profibrotic TGFβ signaling, TGFβ stimulated-human lung fibroblast-derived PD-L1 into extracellular vesicles (EVs) capable of inhibiting T cell proliferation in response to T cell receptor stimulation and mediating fibroblast cell migration. These findings provide new insights and potential targets for a variety of fibrotic and malignant diseases.

This study investigated whether and how classroom-level bullies’ friendship networks, which reflect the popularity of bullies, were associated with the self-esteem of middle school students in the classroom. We analyzed survey data from 2,444 students in 101 classrooms from 22 middle schools in Korea. Respondents reported their friends’ and bullies’ names from their classroom. For each student, we generated the number of close friends that a respondent reported (outdegree) and the number of classmates who nominated him or her as a close friend (indegree). Bullies’ popularity was measured by the sum of all the bullies’ indegrees in a classroom. The findings showed that students in a classroom of bullies with larger popularity had a lower level of self-esteem. Moreover, bullies’ popularity was harmful to nonvictims’ self-esteem as well as victims’ self-esteem. This study suggests that even indirect exposure to bullying in a classroom can decrease the self-esteem of nonvictims by leading to negative emotions such as anger, anxiety, fear, and/or guilty feelings.

It is well known that marital status is significantly associated with mortality risk. Little is known, however, regarding whether and how the effects of marital status are moderated by one's own family structure in childhood. The purposes of this study are to examine whether marital status (i.e., family structure in adulthood) and living with both biological parents in childhood (i.e., family structure in childhood) are associated with mortality risk, and whether and how the effects of marital status vary depending on family structure in childhood and gender. We analyze the risk of death in five waves of the General Social Survey (GSS) from 1994 through 2002 after linking the GSS data to death certificate data from the National Death Index through 2008. The findings indicate that being widowed increases the risk of mortality, while living with both parents in childhood lowers it. Interestingly, analysis of the interaction between marital status and family structure in childhood reveals that the disadvantage of widowhood in terms of mortality is significantly stronger for those who lived with both parents in childhood than for those who did not. Subsample analysis by gender shows that the moderating effect of living with both parents is largely equal across men and women, though statistically more robust for men. These findings suggest that living with both parents during childhood may increase vulnerability to marital disruptions due to unwanted life events such as spousal loss. Childhood advantages, ironically, may form more stressful contexts of spousal loss by lowering one's adaptability or immunity to adulthood hardships, especially when the hardships in adulthood are characteristically opposite from the childhood advantages.

We evaluated the clinical course and significance of postoperative subdural fluid collection (SFC) and identified the patients who were at risk of developing postoperative chronic subdural hematoma (CSDH) after the clipping of unruptured intracranial aneurysms (UIAs).Between January 2012 and June 2014, we retrospectively reviewed 298 patients with UIAs treated by microsurgical clipping. Among them, 257 patients were enrolled in the present study. Subdural lesions (SDLs) were defined as SFC at 1-month follow-up computed tomography (CT) and a CSDH at any time within 1 month after the clipping of UIAs. We examined the volume changes, Hounsfield unit (HU) values, and the end results of SFC in serial CT scans.The incidence of postoperative CSDH that needed burr hole surgery was 2.5%. Changes in SFC volume that occurred within 1 week of surgery were a risk factor for the occurrence of SDL at the 1-month follow-up CT (odds ratio 34.039; P0.001). The corrected average HU value of SCF (cut-off value: 11.9, with a sensitivity of 83.3% and specificity of 73.7%) on postoperative day 7 was an independent risk factor for development of a CSDH at the 1-month follow-up CT (odds ratio 19.261; P = 0.003).SDLs seen during 1-month follow-up may be associated with the occurrence of increased SFC volume within a week after the clipping of UIAs. The corrected average HU value of the SFC on postoperative day 7 was the only risk factor for the development of CSDHs at 1-month follow-up CT.

Sorting nexin 9 (SNX9) is shown to differentiate Smad3 from Smad2 nuclear delivery by mediating the association of phosphorylated Smad3 with importin 8 and the nuclear membrane. While the absence of SNX9 had negligible effects on transforming growth factor β receptor activity or Smad2 signaling, Smad3-dependent targets and phenotypes were inhibited., Transforming growth factor β (TGFβ) is a pleiotropic protein secreted from essentially all cell types and primary tissues. While TGFβ’s actions reflect the activity of a number of signaling networks, the primary mediator of TGFβ responses are the Smad proteins. Following receptor activation, these cytoplasmic proteins form hetero-oligomeric complexes that translocate to the nucleus and affect gene transcription. Here, through biological, biochemical, and immunofluorescence approaches, sorting nexin 9 (SNX9) is identified as being required for Smad3-dependent responses. SNX9 interacts with phosphorylated (p) Smad3 independent of Smad2 or Smad4 and promotes more rapid nuclear delivery than that observed independent of ligand. Although SNX9 does not bind nucleoporins Nup153 or Nup214 or some β importins (Imp7 or Impβ), it mediates the association of pSmad3 with Imp8 and the nuclear membrane. This facilitates nuclear translocation of pSmad3 but not SNX9.

A novel motif within the cytoplasmic tail of the type I TGF-β receptor (TβRI) controls basolateral delivery. While this element functions independent of TβRI recycling and heteromeric TGF-β receptor trafficking, it can dominantly direct an apically expressed receptor to the basolateral membrane in polarized epithelial cells., Delivery of biomolecules to the correct subcellular locales is critical for proper physiological function. To that end, we have previously determined that type I and II transforming growth factor beta (TGF-β) receptors (TβRI and TβRII, respectively) localize to the basolateral domain in polarized epithelia. While TβRII targeting was shown to be regulated by sequences between amino acids 529 and 538, the analogous region(s) within TβRI is unknown. To address that question, sequential cytoplasmic TβRI truncations and point mutations identified a targeting motif between residues 158 and 163 (VxxEED) required for basolateral TβRI expression. Further studies documented that receptor internalization, down-regulation, direct recycling, or Smad signaling were unaffected by motif mutations that caused TβRI mislocalization. However, inclusion of amino acids 148–217 containing the targeting motif was able to direct basolateral expression of the apically sorted nerve growth factor receptor (NGFR, p75; extracellular and transmembrane regions) in a dominant manner. Finally, coexpression of apically targeted type I and type II TGF-β receptors mediated Smad3 signaling from the apical membrane of polarized epithelial cells. These findings demonstrate that the absence of apical TGF-β signaling in normal epithelia is primarily a reflection of domain-specific receptor expression and not an inability to couple with the signaling machinery.

Very few studies have examined the effects of both religious affiliation and religiosity on mortality at the same time, and studies employing multiple dimensions of religiosity other than religious attendance are rare. Using the newly created General Social Survey-National Death Index data, our report contributes to the religion and mortality literature by examining religious affiliation and religiosity at the same time. Compared to Mainline Protestants, Catholics, Jews, and other religious groups have lower risk of death, but Black Protestants, Evangelical Protestants, and even those with no religious affiliation are not different from Mainline Protestants. While our study is consistent with previous findings that religious attendance leads to a reduction in mortality, we did not find other religious measures, such as strength of religious affiliation, frequency of praying, belief in an afterlife, and belief in God to be associated with mortality. We also find interaction effects between religious affiliation and attendance. The lowest mortality of Jews and other religious groups is more apparent for those with lower religious attendance. Thus, our result may emphasize the need for other research to focus on the effects of religious group and religious attendance on mortality at the same time.

NR4A1 (Nur77, TR3) is an orphan nuclear receptor that is overexpressed in pancreatic cancer and exhibits pro-oncogenic activity. RNA interference of NR4A1 expression in Panc-1 cells induced apoptosis and subsequent proteomic analysis revealed the induction of several markers of endoplasmic reticulum stress, including glucose-related protein 78 (GRP78), CCAAT/enhancer-binding protein-homologous protein (CHOP), and activating transcription factor-4 (ATF-4). Treatment of pancreatic cancer cells with the NR4A1 antagonist 1,1-bis(3′-indolyl)-1-(p-hydroxyphenyl)methane (DIM-C-pPhOH) gave similar results. Moreover, both NR4A1 knockdown and DIM-C-pPhOH induced reactive oxygen species (ROS), and induction of ROS and endoplasmic reticulum stress by these agents was attenuated after cotreatment with antioxidants. Manipulation of NR4A1 expression coupled with gene expression profiling identified a number of ROS metabolism transcripts regulated by NR4A1. Knockdown of one of these transcripts, thioredoxin domain containing 5 (TXNDC5), recapitulated the elevated ROS and endoplasmic reticulum stress; thus, demonstrating that NR4A1 regulates levels of endoplasmic reticulum stress and ROS in pancreatic cancer cells to facilitate cell proliferation and survival. Finally, inactivation of NR4A1 by knockdown or DIM-C-pPhOH decreased TXNDC5, resulting in activation of the ROS/endoplasmic reticulum stress and proapoptotic pathways. Implications: The NR4A1 receptor is pro-oncogenic, regulates the ROS/endoplasmic reticulum stress pathways, and inactivation of the receptor represents a novel pathway for inducing cell death in pancreatic cancer. Mol Cancer Res; 12(4); 527–38. ©2014 AACR.

B7-1 proteins are routinely expressed on the surface of antigen presenting cells (APC) and within the innate immune system. They function to establish a biologically optimal and dynamic balance between immune activation and inhibition or self-tolerance. Interactions between B7-1 and its receptors, which include CD28, CTLA4 and PD-L1, contribute to both stimulatory as well as inhibitory or homeostatic regulation. In the current study, we investigated whether the tumor-promoting actions of transforming growth factor beta (TGF-β) disrupted this equilibrium in pancreatic cancer to promote malignant progression and an enhanced means to evade immune detection. The data show that B7-1 is (i) upregulated following treatment of pancreatic carcinoma cells with TGF-β; (ii) induced by TGF-β via both Smad2/3-dependent and independent pathways; (iii) required for pancreatic tumor cell in vitro migration/invasion; and (iv) necessary for TGF-β regulated epithelial-mesenchymal transition (EMT) through induction of Snail family members. Results from the proposed studies provide valuable insights into mechanisms whereby TGF-β regulates both the innate immune response and intrinsic properties of pancreatic tumor growth.

CTGF (connective tissue growth factor) is widely regarded as an important amplifier of the profibrogenic action of TGF-β (transforming growth factor β) in a variety of tissues, although the precise mechanism of how the TGF-β signalling pathways modulate CTGF expression remains unclear. In the present study, the role of PKCδ (protein kinase Cδ) in TGF-β1-mediated CTGF expression was investigated using HepG2 cells. TGF-β1 treatment specifically elevated PKCδ activation and CTGF expression. In contrast, blockade of PKCδ by the selective inhibitor Rottlerin or by siRNA knockdown significantly reduced TGF-β1-induced CTGF production. The regulatory mechanism was further demonstrated in HepG2 cells whereby TGF-β1-induced PKCδ activation negatively regulated the nuclear levels of PPM1A (protein phosphatase, Mg2+/Mn2+ dependent, 1A) through the RhoA/ROCK (Rho-associated kinase) pathway. Moreover, we showed that both Smad signalling and the PKCδ pathway appeared to be stimulated by TGF-β1 in parallel. Time course assessments indicated that PKCδ signalling may have a function in maintaining nuclear phospho-Smads at a maximal level. The collective results of the present study demonstrated that PKCδ-stimulated RhoA/ROCK activation resulted in a reduction in PPM1A, thereby up-regulating Smad-dependent gene induction for extended periods. These findings indicated that PKCδ plays a critical role in TGF-β1-induced CTGF production in HepG2 cells.

IL-33 is elevated in afflicted tissues of patients with mast cell (MC)–dependent chronic allergic diseases. Based on its acute effects on mouse MCs, IL-33 is thought to play a role in the pathogenesis of allergic disease through MC activation. However, the manifestations of prolonged IL-33 exposure on human MC function, which best reflect the conditions associated with chronic allergic disease, are unknown. In this study, we found that long-term exposure of human and mouse MCs to IL-33 results in a substantial reduction of MC activation in response to Ag. This reduction required >72 h exposure to IL-33 for onset and 1–2 wk for reversion following IL-33 removal. This hyporesponsive phenotype was determined to be a consequence of MyD88-dependent attenuation of signaling processes necessary for MC activation, including Ag-mediated calcium mobilization and cytoskeletal reorganization, potentially as a consequence of downregulation of the expression of phospholipase Cγ1 and Hck. These findings suggest that IL-33 may play a protective, rather than a causative, role in MC activation under chronic conditions and, furthermore, reveal regulated plasticity in the MC activation phenotype. The ability to downregulate MC activation in this manner may provide alternative approaches for treatment of MC-driven disease.

Macroautophagy is involved in the bulk degradation of long-lived cytosolic proteins and subcellular organelles, which is important for the survival of cells during starvation. To identify potential players of the autophagy process, we subjected HCT116 cells cultured in complete medium and in Earle's balanced salt solution to proteomics analysis. In approximately 1500 protein spots detected, we characterized 52 unique proteins, whose expression levels were significantly changed following starvation. Notably, we found that Annexin A1 was significantly upregulated following starvation at both mRNA and protein levels. Inhibition of Annexin A1 expression with specific siRNA did not alter starvation-induced autophagy as measured by the level of lipidated LC3, but significantly reversed autophagy degradation as measured by the level of p62/SQSTM 1. Thus Annexin A1 seemed to be positively upregulated during starvation to promote autophagic degradation. Overall, the data presented in this study established a expression profile of the proteome in starved cells, which allowed the identification of proteins with potential significance in starvation-induced autophagy.

Chemopreventive or anticancer agents induce cancer cells to apoptosis through the activation of adenosine AMP-activated protein kinase (AMPK), which plays a major role as energy sensors under ATP-deprived condition or ROS generation. In this study, we compared the effects of ascochlorin (ASC), from the fungus Ascochyta viciae, and its derivatives on AMPK activity. We also examined a regulatory mechanism for hypoxia-inducible factor-1α (HIF-1α) stabilization in response to 4-O-methylascochlorin (MAC). We found that AMPK activation was mainly involved with MAC, but not ASC and 4-O-carboxymethylascochlorin (AS-6), indicating that the substitution of 4-O-methyl group from 4-O-hydroxyl group of ASC is important in the activation of AMPK and the expression of HIF-1α. MAC-stabilized HIF-1α via AMPK activation triggered by lowering the intracellular ATP level, not by ROS generation, increases glucose uptake and the expression of vascular endothelial growth factor (VEGF) and glucose transporter 1 (GLUT-1), major target genes of HIF-1α. Moreover, MAC-induced AMPK activity suppressed survival factors, including mTOR and ERK1/2 or translational regulators, including p70S6K and 4E-BP1. Our data suggest that AMPK is a key determinant of MAC-induced HIF-1α expression in response to energy stress, further implying its involvement in MAC-induced apoptosis.

The relationship between lower urinary tract symptoms and depressive symptoms was assessed using data from the Male Attitudes Regarding Sexual Health study.Lower urinary tract symptoms, depressive symptoms and erectile dysfunction were assessed using International Prostate Symptom Score, Center for Epidemiologic Studies Depression Scale and a validated question from the Massachusetts Male Aging Study. Sociodemographic, clinical and other data were also collected. Odds ratios and 95% CIs were determined using weighted multivariate logistic regression stratified by race/ethnicity and age.Of 3,291 randomly selected men 2,173 completed the interview. Overall odds of lower urinary tract symptoms were increased in men who reported depressive symptoms (OR 2.68, 95% CI 1.60-4.47, p0.01), erectile dysfunction (OR 1.73, 95% CI 1.11-2.71, p0.05) and unhappiness/dissatisfaction on the International Prostate Symptom Score quality of life item (OR 10.72, 95% CI 5.56-20.69, p0.01), and those 60 to 69 years old (OR 1.99, 95% CI 1.14-3.46, p0.05) and 70 years or older (OR 1.91, 95% CI 1.06-3.43, p0.05). Increased odds of lower urinary tract symptoms were associated with depressive symptoms for white (OR 2.60, 95% CI 1.39-4.85, p0.01) and Hispanic men (OR 4.14, 95% CI 1.15-14.95, p0.05). Odds of depressive symptoms were increased in men reporting lower urinary tract symptoms (OR 2.64, 95% CI 1.57-4.43, p0.001), especially Hispanic men 50 to 59 years old (OR 133.17, 95% CI 18.40-963.87, p0.01) and black men older than 60 years (OR 21.61, 95% CI 3.04-153.55, p0.01), as well as men 40 to 49 years old expressing unhappiness/dissatisfaction on the International Prostate Symptom Score quality of life item (OR 6.80, 95% CI 1.77-26.16, p0.01), and Hispanic (OR 10.76, 95% CI 3.88-29.80, p0.01) and black men reporting erectile dysfunction (OR 4.77, 95% CI 1.15-19.78, p0.05), but not white men reporting erectile dysfunction (OR 1.05, 95% CI 0.48-2.28, p0.91).Lower urinary tract symptoms and depressive symptoms are strongly associated, and exhibit reciprocal relationships. Erectile dysfunction increases the odds of both disorders.

Objectives. Faced with aging societies, there is an immense need to better understand the nature of volunteering outside advanced Western industrial countries. As a case of a rapidly aging society, we identify robust factors associated with elderly volunteering in Korea in terms of a resource framework. Methods. Data were derived from the Social Statistics Survey conducted by the Korea National Statistical Office in 1999 (N = 7,135) and 2003 (N=8,371). We first determined overall and age-related volunteer rates for Korea compared to the United States. Using logistic regression, we then examined the effects of human, cultural, and social capital variables on volunteering. Results. Approximately 6% of Koreans aged 65 years and older participate in volunteer programs. All human capital variables are positively related with volunteering. For cultural capital, those who identify their religion as Buddhism or Catholicism are more likely to volunteer than those who have no religion. But surprisingly, Protestantism does not consistently promote volunteering across both years. For social capital, older adults who live alone or with a spouse are more likely to volunteer than those living with both a spouse and children. Discussion. In contrast to human capital, cultural and social capital on elderly volunteering appears to be contoured by social contexts.

The expression of matrix metalloproteinase-9 (MMP-9) has been implicated in the invasion and metastasis of cancer cells. Here, we found that an antitumor antibiotic, ascofuranone, inhibits invasion and MMP-9 induction induced by phorbol myristate acetate (PMA) in human cell lines. Ascofuranone also inhibits the protein expression and transcription of MMP-9 induced by tumor necrosis factor-alpha. The inhibition of MMP-9 induction was observed in human cancer cell lines as well as primary rat mesangial cells. Furthermore, as evidenced by MMP-9 promoter and electrophoretic mobility shift assays, ascofuranone specifically inhibited MMP-9 gene expression by blocking PMA-stimulated activation of activator protein-1 (AP-1). In addition, ascofuranone suppressed PMA-induced phosphorylation of Ras, Raf, MEK and extracellular signal-regulated kinase (ERK), upstream factors involved in AP-1activation, whereas the phosphorylation of p38 and JNK/mitogen-activated protein kinase was not affected by ascofuranone, suggesting that the primary target of ascofuranone for suppression of the AP-1 induction is present in upstream of ERK signaling pathway. These results suggest that the suppression of MMP-9 expression, at least in part, contributes to the antitumor activity of ascofuranone.

Ascochlorin is a prenyl-phenol compound that was isolated from the fungus Ascochyta viciae. Ascochlorin reduces serum cholesterol and triglyceride levels, suppresses hypertension and tumor development, and ameliorates type I and II diabetes. Here, to better understand the mechanisms by which ascochlorin regulates physiological or pathological events and induces responses in the pharmacological treatment of cancer, we performed differential analysis of the proteome of the human osteosarcoma cells U2OS in response to ascochlorin. In addition, we established the first two-dimensional map of the U2OS proteome. The U2OS cell proteomes with and without treatment with ascochlorin were compared using two-dimensional electrophoresis, matrix-assisted laser desorption/ionization mass spectrometry and bioinformatics. The largest differences in expression were observed for the epidermal growth factor receptor (4-fold decrease), ribulose-5-phosphate-epimerase (13-fold decrease), ATP-dependent RNA helicase (8-fold decrease), and kelch-like ECH-associated protein 1 (6-fold decrease). The abundance of heterogeneous nuclear ribonucleoprotein L and minichromosome maintenance protein 7 increased 12- and 8.2-fold, respectively. In addition, Erk 2 was increased 3-fold in U2OS cells treated with ascochlorin. The expression of some selected proteins was confirmed by western blotting, zymography and RT-PCR analysis.

Subjective sexual well-being refers to the cognitive and emotional evaluation of an individual's sexuality. This study examined subjective sexual well-being, explored its various aspects, examined predictors across different cultures, and investigated its possible associations with overall happiness and selected correlates, including sexual dysfunction. Data were drawn from the Global Study of Sexual Attitudes and Behaviors, a survey of 27,500 men and women aged 40-80 years in 29 countries. The cross-national variation of four aspects of sexual well-being (the emotional and physical satisfaction of sexual relationships, satisfaction with sexual health or function, and the importance of sex in one's life) was explored using cluster analysis, and relationships among sexual well-being, general happiness, and various correlates were examined using ordinary least squares regression and ordered logistic regression. Results from the cluster analysis identified three clusters: a gender-equal regime and two male-centered regimes. Despite this cultural variation, the predictors of subjective sexual well-being were found to be largely consistent across world regions.

Native low-density lipoprotein (LDL) and oxidized LDL (oxLDL) possess a wide variety of biological properties, and play a central role in atherogenesis. In this study, we used a proteomic analysis of human monocyte THP-1 cells induced with oxLDL or with LDL, to identify proteins potentially involved in atherosclerotic processes. Of the 2500 proteins detected, 93 were differentially expressed as a result of priming with LDL or oxLDL. The proteins were unambiguously identified by comparing the masses of their tryptic peptides with those of all known proteins using MALDI-TOF MS and the NCBI database. The largest differences in expression were observed for vimentin (94-fold increase), meningioma-expressed antigen 6 (48-fold increase), serine/threonine protein phosphatase 2A (40-fold increase), and beta-1,3-galactosyltransferase (15-fold increase). In contrast, the abundance of an unnamed protein product and phosphogluconate dehydrogenase decreased 30-fold and 25-fold, respectively. The expression of some selected proteins was confirmed by Western blot and RT-PCR analyses. The proteins identified in this study are attractive candidates for further biomarker research. This description of the altered protein profiles induced by oxLDL in human monocytes will support functional studies of the macrophage-derived foam cells involved in the pathogenesis of atherosclerosis.

The succinic semialdehyde dehydrogenase gene (SSADH; EC 1.2.1.24) from human brain was cloned and overexpressed in Escherichia coli . Based on SDS–PAGE, the apparent molecular mass of subunit was 54 kDa, in good agreement with the theoretical size. The purified SSADH appears to be a tetramer of identical subunits. The specific activity of the recombinant protein was 1.82 μmol NADH formed min −1 mg −1 and the optimal pH was found to be 8.5. The Michaelis constants K m for succinic semialdehyde and NAD + were 6.3 and 125 μM, respectively. Initial velocity studies show NADH to be a competitive inhibitor with respect to NAD + , but to be non-competitive inhibitor with respect to succinic semialdehyde. The overexpression of SSADH in E. coli and one-step purification of the highly active SSADH will facilitate further biochemical studies on this enzyme. In addition, an mRNA master dot-blot for multiple human tissues provided a complete map of the tissue distribution for SSADH. The major sites of SSADH expression are liver, skeletal muscle, kidney, and brain. The data indicate that mRNA expression of SSADH is ubiquitous, but highly regulated at the level of transcription in a tissue-specific manner.

We used a combined computer and biochemical approach to characterize human pyridoxine 5'-phosphate oxidase (PNPO). The human PNPO gene is composed of seven exons and six introns, and spans approximately 8 kb. All exon/intron junctions contain the gt/ag consensus splicing site. The absence of TATA-like sequences, the presence of Sp1-binding sites and more importantly, the presence of CpG islands in the regulatory region of the PNPO gene are characteristic features of housekeeping genes. Northern blot analyses showed two species of poly(A)(+) RNA of approximately 2.4 and approximately 3.4 kb at identical intensity, whereas Western blot analysis showed that no protein isoform exists in any of the tissues examined. PCR-based analysis led to the idea that two messages are transcribed from a single copy gene, and that the size difference is due to differential usage of the polyadenylation signal. The major sites of PNPO expression are liver, skeletal muscle and kidneys while a very weak signal was detected in lung. The mRNA master dot-blot for multiple human tissues provided a complete map of the tissue distribution not only for PNPO but also for pyridoxal kinase and pyridoxal phosphatase. The data indicate that mRNA expression of all three enzymes essential for vitamin B(6) metabolism is ubiquitous but is highly regulated at the level of transcription in a tissue-specific manner. In addition, human brain PNPO cDNA was expressed in Escherichia coli, and the roles of both the N- and C-terminal regions were studied by creating sequential truncation mutants. Our results showed that deletion of the N-terminal 56 residues affects neither the binding of coenzyme nor catalytic activity.

Objectives: The purpose of this study was to clarify the prognostic significance of visceral pleura invasion in T2 non-small cell lung cancer (NSCLC). Materials and methods: Between 1990 and 2001, 439 consecutive patients with T2 NSCLC underwent curative surgical resection. The subjects included 234 patients with stage IB, 95 with stage IIB, and 110 with stage IIIA and B disease. The patients were divided into two groups according to the existence of visceral pleura invasion (group I without, group II with). Both groups were compared with regard to tumor size, histology, associated mediastinal lymph node involvement, and survival rates. Results: Visceral pleura invasion (group II) was identified in 114 patients (26%), and was present in 22% of patients with NSCLC with a tumor size of 3 cm or less and in 27% of those with a tumor larger than 3 cm (P ¼ 0:37). Visceral pleura invasion was associated with a higher frequency of mediastinal lymph node involvement (group I ¼ 22%, group II ¼ 34%, P ¼ 0:009). Five- and 10-year survival rates were 50 and 45% in group I, and 36 and 22% in group II (P ¼ 0:0006). In stage IB, visceral pleura invasion was identified in 53 patients (23%), and 5- and 10-year survival rates were 63 and 60% in the visceral pleura non-invasion group, and 44 and 28% in visceral pleura invasion group (P ¼ 0:0018). By multivariate Cox model analysis, age at intervention (relative risk ¼ 1.03, P ¼ 0:0017), N status (relative risk ¼ 1.53, P , 0:0001), tumor size (relative risk ¼ 1.83, P ¼ 0:0452) and visceral pleura invasion (relative risk ¼ 1.42, P ¼ 0:0291) were independent predictors of poor prognosis. Conclusions: We were able to demonstrate that visceral pleura invasion was a factor of poor prognosis in T2 NSCLC. It was found to correlate with more extensive mediastinal lymph node involvement and a decreased survival rates. Therefore, the patients with visceral pleura invasion should be closely followed up especially. q 2003 Elsevier Science B.V. All rights reserved.

Autophagy can be activated via MTORC1 down-regulation by amino acid deprivation and by certain chemicals such as rapamycin, torin, and niclosamide. Lysosome is the degrading machine for autophagy but has also been linked to MTORC1 activation through the Rag/RRAG GTPase pathway. This association raises the question of whether lysosome can be involved in the initiation of autophagy. Toward this end, we found that niclosamide, an MTORC1 inhibitor, was able to inhibit lysosome degradation and increase lysosomal permeability. Niclosamide was ineffective in inhibiting MTORC1 in cells expressing constitutively activated Rag proteins, suggesting that its inhibitory effects were targeted to the Rag-MTORC1 signaling system. This places niclosamide in the same category of bafilomycin A1 and concanamycin A, inhibitors of the vacuolar H(+)-ATPase, for its dependence on Rag GTPase in suppression of MTORC1. Surprisingly, classical lysosome inhibitors such as chloroquine, E64D, and pepstatin A were also able to inhibit MTORC1 in a Rag-dependent manner. These lysosome inhibitors were able to activate early autophagy events represented by ATG16L1 and ATG12 puncta formation. Our work established a link between the functional status of the lysosome in general to the Rag-MTORC1 signaling axis and autophagy activation. Thus, the lysosome is not only required for autophagic degradation but also affects autophagy activation. Lysosome inhibitors can have a dual effect in suppressing autophagy degradation and in initiating autophagy.

The synthetic derivative of ascochlorin, 4-O-carboxymethyl ascochlorin (AS-6) is an agonist of the nuclear hormone receptor PPARγ and has been shown to induce differentiation in mouse pre-adipocytes and to ameliorate type II diabetes in a murine model. AS-6 was cytotoxic when added at micromolar concentrations to cultures of three different human cancer cell lines. We used gel electrophoresis and mass spectrometry to identify proteins with altered expression in human hepatocarcinoma cells (HepG2) cells after 12 h in the presence of AS-6 and found 58 proteins that were differentially expressed. Many of the proteins showing increased expression in cells treated with AS-6 are involved in protein quality control, including glucose-regulated protein 78 (GRP78/BiP), a regulator of ER stress responses, and the transcriptional regulator CHOP, which mediates ER stress-induced apoptosis. Cells treated with AS-6 undergo an autophagic response accompanied by increased expression of beclin1, ATG5, and LC3-II and autophagosome formation marked by the appearance of large vesicles containing LC3-II. Grp78 induction was inhibited when the PPARγ antagonist, GW9662, was added together with AS-6, and autophagy and cell death were partially blocked. 3-methyl-adenine (3-MA), an inhibitor of phosphatidyl inositol 3-kinase (PI3-kinase) prevented induction of ATG5 and activation of LC3-II and blocked autophagosome formation. 3-MA also blocked induction of GRP78 and CHOP, suggesting that PI3-kinase, which is known to mediate ER stress-induced autophagy, also plays a role in initiating apoptosis in response to ER stress. Together these data establish that the cytotoxicity of AS-6 operates by a mechanism dependent on ER stress-induced autophagy and apoptosis.

Obesity causes changes in fatty acid metabolism that consequently leads to fatty liver. To identify the possible proteins involved in the processes of obesity, we performed a proteomic analysis of obesity-induced mouse liver. Male C57BL/6J mice that were fed a high-fat diet (HFD) for 24wk, developed hepatic steatosis characterized by considerable increase in free fatty acid (FFA) and triglyceride levels. Body weights were measured weekly and other measurements at weeks 2, 6, 12, 16, and 24. 2-D-based proteomic analysis revealed that, compared with the normal diet (ND) (n 550), high-fat diet (n 550) changed the expression of 12 protein (8 up and 4 downregulated, by a 1.5 fold change and more, po0.05). The most pronounced difference was observed in intermediate microfilament (IF) cytoskeleton proteins. In particular, vimentin (vim) as well as cytokeratins (CK-8 and CK-18) were significantly upregulated in obese animals. Moreover, the level of caspase-generated IF fragment was also positively correlated with the degree of steatosis. The results suggest a significant alteration in IF organization during the development of hepatic steatosis leading to inflammation. The expression profile of selected proteins including vim was validated by Western blot, microarray analysis, and hepatocyte morphology by immunohistochemistry. Our results suggest that vim, like CK-18, may be a useful marker for predicting obesity and liver disease.

Background: The purpose of this study was to introduce a combination procedure with double C-shaped skin incision and an adjusted dual-floor burr hole to prevent skin complications on the scalp with deep brain stimulation (DBS) surgery. Methods: Between March 2000 and March 2010, 504 DBS electrodes were implanted in 268 patients. We included both bilateral and unilateral DBS surgery cases for Parkinson’s disease, tremor, pain, obsessive-compulsive disorder and intractable seizure accompanied with cranial and IPG insertion procedure, but excluded motor cortex stimulation, and spinal cord stimulation. We used a straight skin incision in 118 patients, double C-shaped skin incision only in 113 patients since March 2006, and combined a double C-shaped skin incision and dual-floor burr hole in 37 patients since August 2009. We compared scalp wound complications and the height subscale of the Vancouver Scar Scale between previous straight or C-shaped skin incision and the combination procedure. Results: We had eight scalp erosions associated with infection (3.0%) and six disconnection (2.2%) cases among 268 patients. Before the use of the double C- skin incision or adjusted dual-floor burr hole technique, we had 5 (4.2%) scalp erosion cases among 118 patients. With the introduction of the C-shaped incision, only 3 patients (2.7%) developed scalp erosion. However, no patient among 37 patients developed scalp erosion after using both double C-shaped skin incision and dual-floor burr hole technique. Scalp bump measured by the height subscale of the Vancouver Scar Scale was more cosmetic in the combination procedure. Conclusion: The combination procedure prevents skin complications associated with DBS surgery, with tensile strength, less impaired vascular supply and better cosmetic outcome. This promising approach prevents unwanted skin complications associated with DBS surgery and improves patient satisfaction.

Autophagosomes and their precursors are best defined by electron microscopy but may also be traced in living cells based on the distribution of specific autophagy molecules. LC3, the most commonly examined autophagy marker in mammalian cells, labels structures that are frequently manifested as dots or rings using light microscopy; however, the nature of these structures is not entirely clear. We reported here a novel approach to examine the LC3-positive compartment in cell-free lysates, which revealed that they were actually tubulovesicular structures with considerable heterogeneity. Using affinity purification, we isolated these membranes for electron microscopy, which indicated that they possessed ultrastructural features consistent with autophagosomal membranes at various maturation stages. Further biochemical and proteomics analyses demonstrated the presence of multiple autophagy-related and other functional molecules. The different distribution patterns of Atg5, Atg16, Atg9, and p62/SQSTM1 on the LC3-positive compartment provided new clues on how these molecules might be involved in the dynamics of the autophagosomal membranes. Finally, several morphologically unique groups of LC3-positive membranes were categorized. Their topological configurations suggested that double-membrane vesicles could be derived from single membrane compartments via different means, including tubule-to-vesicle conversion, whose presence was supported by live cell imaging. These findings thus provide new information on the dynamics of the autophagosomal compartment.

The proteasome and the autophagy systems are two evolutionarily conserved mechanisms for degrading intracellular materials. They are functionally coupled and suppression of the proteasome promotes autophagy. Although suppression of the proteasome leads to cell death, suppression of autophagy can be either prodeath or prosurvival. To understand the underlining mechanism of this dichotomy and its potential clinical implications, we treated various transformed and nontransformed human cells with proteasome inhibitors. We found that whether autophagy served a prosurvival role in this scenario was contingent on the cellular oncogenic status. Thus, autophagy suppression enhanced apoptosis induced by proteasome inhibitors in transformed cells, but not in nontransformed cells. Oncogenic transformation enhanced the ability of cells to initiate autophagy in response to stress, reflecting a stronger dependence of transformed cells on autophagy for survival. Indeed, a combined use of bortezomib, the only Food and Drug Administration–approved proteasome inhibitor for clinical use, and chloroquine, which inhibits autophagy by disturbing lysosomal functions, suppressed tumor growth more significantly than either agent alone in a xenograft model. These findings indicate that suppression of both intracellular degradation systems could constitute a novel strategy for enhanced cancer control in a tumor-specific way. [Mol Cancer Ther 2009;8(7):2036–45]

Proteomics is a novel molecular profiling technology. It is mainly concerned with determining the structure, expression, localization, biochemical activity, interactions, and cellular roles of any proteins. Clinical research hopes to benefit from proteomics in the identification of new drug targets and the development of new diagnostic markers. Better to understand the mechanisms by which ascofuranone (AF), an isoprenoid antibiotic, regulates physiological or pathological events and induces responses in the pharmacological treatment of cancer, we performed differential analysis of the human osteosarcoma cells U2OS proteomes in response to this agent. The U2OS cell proteomes with and without treatment with AF were compared using two-dimensional electrophoresis, matrix-assisted laser desorption/ionization mass spectrometry, and bioinformatics. The largest differences in protein expression were observed for hydroxyindole O-methyltransferase, syntaxin-binding protein 1, the matrix metalloproteinase (MMP)-2, urokinase receptor, and endothelial protein C receptor. Changes in expression and activity of some selected proteins were confirmed by Western blotting, zymography, and reverse transcription-polymerase chain reaction analysis. In particular, we observed downregulated tumor growth related-proteins such as MMP-2 and endothelial protein C receptor. According to these results, AF might be useful as a potent chemotherapeutic agent.

The invasion of monocytes through the endothelial wall of arteries and their transformation from macrophage into form cells has been implicated as a critical initiating event in atherogenesis. Human THP-1 monocytic cells can be induced to differentiate into macrophages by phorbol myristate acetate (PMA) treatment, and can be converted into foam cells by exposure to oxidized low-density lipoprotein (oxLDL). To identify proteins potentially involved in atherosclerotic processes, we performed a proteomic analysis of THP-1 macrophages exposed to oxLDL generated by treatment with native LDL with hypochlorous acid/hypochlorite (HOCl/OCl(-)). We detected more than a thousand proteins, of which 104 differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) and the NCBI database. The largest differences in expression were observed for bifunctional purine biosynthesis protein, vacuolar protein sorting 33A, breast carcinoma amplified sequence, adenine phosphoribosyltransferase, and tropomyosin alpha 3 chain. Interestingly, many apoptotic proteins such as lamin B1, poly (ADP-ribose) polymerase, Bcl-2 related protein A1 and vimentin were identified by MALDI-TOF analysis. Identities were confirmed by matching the sequence of several tryptic peptides using MALDI-TOF/TOF MS, Western blot analyses and immunofluorescent microscopy. The data described here will contribute to establishing a functional profile of the human macrophage proteome. Furthermore, the proteins identified in this study are attractive candidates for further biomarkers involved in the pathogenesis of atherosclerosis.

Summary Aims: Lower urinary tract symptoms (LUTS) are categorised as storage (urgency, frequency, nocturia and incontinence), voiding (sensation of incomplete emptying, hesitancy, weak stream and straining) or mixed symptoms. Methods: In this US population-based study, we investigated the prevalence of male LUTS and the relative frequency of the LUTS subtypes, and we evaluated associations between LUTS and age, race/ethnicity and erectile dysfunction (ED). The Male Attitudes Regarding Sexual Health study included a nationally representative sample of non-Hispanic black, non-Hispanic white and Hispanic men aged ≥ 40 years. Participants completed a questionnaire including items on ED and the International Prostate Symptom Score (IPSS). The prevalence and subtypes of LUTS were investigated post hoc by age, race/ethnicity and the presence of ED. Results: The overall prevalence rates of storage (13%) and mixed (9%) symptoms were higher than that of voiding symptoms (6%). The prevalence of storage symptoms was similar across age groups, whereas voiding and mixed symptoms increased with age. Among men with IPSS ≥ 8, the rates of storage (29%) and mixed (38%) symptoms were also higher than voiding symptoms (23%). Distributions of the LUTS subtypes were comparable among black, white and Hispanic respondents with IPSS ≥ 8. The overall prevalence rate of ED (40%) increased with age among those with IPSS ≥ 8. Isolated storage symptoms were more than twice as common as isolated voiding symptoms among US men ≥ 40 years of age. Conclusion: Careful attention to individual symptoms may help distinguish storage LUTS from voiding LUTS, a distinction that has important implications for treatment.

The critical initiating event in atherogenesis involves the invasion of monocytes through the endothelial walls of arteries and the transformation of monocytes from macrophages into foam cells. Human THP-1 monocytic cells can be induced to differentiate into macrophages by phorbol myristate acetate (PMA) and can then be converted into foam cells by exposure to oxidized low-density lipoprotein (oxLDL). Also, during a chronic inflammatory response, monocytes/macrophages produce the 92-kDa matrix metalloproteinase-9 (MMP-9) that may contribute to the extravasation, migration, and tissue remolding capacities of the phagocytic cells. Here, we investigate the effect of ascochlorin (ASC), a prenylphenol antiviral compound from the fungus Ascochyta viciae, on oxLDL-induced MMP-9 expression and activity in human THP-1 macrophages. ASC reduced oxLDL-induced MMP-9 expression and activity in a time-dependent and dose-dependent manner. Also, an analysis of MMP-9 activity using pharmacologic inhibitors showed that ASC inhibits MMP-9 activity via the extracellular signal-regulated kinase 1 and kinase 2 pathways. Our results suggest that ASC may be useful as a potent clinical antiatherogenic agent, a topic of considerable interest in the biological chemistry of chemotherapeutic agents.

Introduction Most U.S. population‐based estimates of erectile dysfunction (ED) prevalence restricted upper age, were not nationally representative, or underrepresented minority groups. Aim To estimate, by race/ethnicity in the United States, the prevalence of ED and the impact of sociodemographic, health, relationship, psychological, and lifestyle variables. Methods This cross‐sectional, population‐based, nationally representative probability survey conducted between May 2001 and January 2002 in the general community setting facilitated equivalent representation among U.S. non‐Hispanic white (N=901), non‐Hispanic black (N=596), and Hispanic (N=676) men aged 40 and older by using targeted phone lists to oversample the minority populations. Main Outcome Measure Estimated prevalence of moderate or severe ED, defined as a response of "sometimes" or "never" to the question "How would you describe your ability to get and keep an erection adequate for satisfactory intercourse?" Results The estimated prevalence was 22.0% (95% confidence interval [CI], 19.4–24.6) overall, 21.9% (95% CI, 18.8–24.9) in whites, 24.4% (95% CI, 18.4–30.5) in blacks, and 19.9% (95% CI, 13.9–25.9) in Hispanics, and increased with increasing age. The odds ratio increased with increasing age. Probability also increased with diabetes, hypertension, and moderate or severe lower urinary tract symptoms (LUTS) overall; age ≥70years and diabetes in whites; severe LUTS in blacks; and age ≥60years, moderate LUTS, hypertension, and depression in Hispanics. It decreased with exercise and college vs. less than high school education overall; with exercise, good relationship quality, and according to alcohol intake in blacks; and with high school or college education in Hispanics. Conclusions The odds of ED increased with increasing age across race/ethnicity when controlling for sociodemographic, health, relationship, psychological, and lifestyle variables. These initial analyses suggest further study of the interrelationships among risk factors for ED. Laumann EO, West S, Glasser D, Carson C, Rosen R, and Kang J‐H. Prevalence and correlates of erectile dysfunction by race and ethnicity among men aged 40 or older in the United States: From the Male Attitudes Regarding Sexual Health survey. J Sex Med 2007;4:57–65.

This study investigated a reference line that is closer to the true femoral anteversion on only the cutting surface of the proximal femoral neck during a femoral stem insertion in a cementless total hip arthroplasty. A postoperative computed tomography of both hips from 33 consecutive patients after a unilateral primary cementless total hip arthroplasty with an avascular necrosis of the femoral head was taken to observe the positioning of the stem and its correlation with the true anteversion of the contralateral side. The average of the midcortical angle was 14.1 degrees +/- 6.8 degrees on the lesion side and 0.1 degrees +/- 1.3 degrees more than the true anteversion on the contralateral side. This study has shown that anteversion using a midcortical line between the anterior cortical line and the posterior cortical line is compatible with the true femoral anteversion. However, further investigation is required to confirm the true femoral anteversion.

The incidence of tuberculosis (Tbc) infection is high in some parts of the world and tuberculous cold abscess of the chest wall (CACW) often fails to respond to medical treatment. Medical records of 178 patients who underwent surgical treatment of chest wall abscesses from July 1970 to Sept. 2000 were reviewed and 89 patients who were pathologically confirmed as Tbc cold abscess cases were included in this study. Their ages ranged from 9 to 71 years (mean 33.3 years) and the male to female ratio was 1.2:1 (49 male, 40 female). The symptoms were palpable chest wall mass, pain and pus discharge, and three patients had multiple lesions. Twenty-five patients (28%) underwent excision of chest wall abscesses and 64 patients (72%) underwent chest wall and rib resection. Tbc medication was given preoperatively in 39 patients for an average of 6.3 months and all patients were given Tbc medication postoperatively for an average of 12 months. Postoperative complications were bleeding, pus discharge, empyema, pleural effusion, wound dehiscence, subcutaneous emphysema and activation of pulmonary Tbc. The disease recurred in 7 patients (7.8%) and these 7 patients all underwent a second operation. We recommend preoperative Tbc medication and complete resection of chest wall abscesses including any suspicious ribs. Postoperative Tbc medication for a minimum of 12 months is essential to decrease the risk of a relapse.