Analysis of severe maternal morbidity  provides information on the quality of care. We assessed the prevalence/incidence of maternal near miss, maternal mortality and case fatality ratio through systematic review of studies on severe maternal morbidity in sub-Saharan Africa.We examined studies that reported prevalence/incidence of severe maternal morbidity  during pregnancy, childbirth and postpartum period between 1996 and 2010. We evaluated the quality of studies . We extracted data, using a pre-defined protocol and criteria, and estimated the prevalence or incidence of maternal near miss. The case-fatality ratios for reported maternal complications were estimated.We identified 12 studies: six were cross-sectional, five were prospective and one was a retrospective review of medical records. There was variation in the setting: while some studies were health facility-based , others were community-based studies. The sample size varied from 557 women to 23,026. Different definitions and terminologies for maternal near miss included acute obstetric complications, severe life threatening obstetric complications and severe obstetric complications. The incidence/prevalence ratio and case-fatality ratio for maternal near misses ranged from 1.1%-10.1% and 3.1%-37.4% respectively. Ruptured uterus, sepsis, obstructed labor and hemorrhage were the commonest morbidities that were analyzed. The incidence/prevalence ratio of hemorrhage ranged from 0.06% to 3.05%, while the case fatality ratio for hemorrhage ranged from 2.8% to 27.3%. The prevalence/incidence ratio for sepsis ranged from 0.03% to 0.7%, while the case fatality ratio ranged from 0.0% to 72.7%.The incidence/prevalence ratio and case fatality ratio of maternal near misses are very high in studies from sub-Saharan Africa. Large differences exist between countries on the prevalence/incidence of maternal near misses. This could be due to different contexts/settings, variation in the criteria used to define the maternal near misses morbidity, or rigor used carrying out the study. Future research on maternal near misses should adopt the WHO recommendation on classification of maternal morbidity and mortality. For every maternal death, there are close to 100 women with severe maternal morbidity referred to as maternal near misses . Comparematernal near miss on this continuum lies in the definition of the threshold of severity above which morbidity qualifies to be a near miss. While this threshold is clear for some obstetric conditions or their management , it is may be uncertain or ambiguous for other conditions (such as sepsis). Secondly, the threshold above which an adverse obstetric event becomes life-threatening may be context specific. This is so because the probability of death from such complications depends not only on the woman's vulnerability to succumb to (or capacity to cope with) a given complication, but also on access to prompt and quality care [maternal near miss have to take the local context into consideration, and therefore health system factors. Three approaches have been proposed for definition of maternal near miss: utilization of clinical features  [The analysis of maternal deaths has long been used for the evaluation of quality of care, women's health and level of socio-economic development. The identification of cases of maternal near misses is an alternative to the investigation of maternal deaths when assessing the quality of obstetric care . Conceptity care ,2,4,5. Trupture) ; criterirupture) , or critWe carried out a systematic review of the literature using the criteria described by Kranke  and MoheFor the assessment of the study quality, a structured data collection form from the WHO systematic review of maternal morbidity and mortality  was usedFigure Twenty studies were excluded from the systematic review, but used in literature review, while 48 studies were excluded entirely for lack of a clear definition of morbidity, unclear study design or unspecific data on the specific obstetric conditions that led to maternal mortality. Most of the 20 studies excluded from the systematic review, but formed background for the literature review, assessed a single or specific maternal morbidity , which was basis for exclusion from the systematic review. Finally, 12 studies were included in the systematic review. The authors of the articles that reported the studies used in this systematic review were contacted and requested to supply more information on the articles. We examined critically the definition used to identify maternal near misses, prevalence or incidence of maternal near miss and case fatality ratio from conditions that constituted maternal near misses.We identified 12 studies which met the criteria for inclusion in the systematic review. Table Table Table This systematic review shows that: 1) Different terminologies are used by different studies to describe women with severe acute maternal morbidity from complications of pregnancy, childbirth or the puerperium. 2) Different studies use different definitions of the maternal near miss. 3) Different studies collect and report differently the information on maternal near miss. 4) Few studies report on the specific causes of morbidity and their contribution to maternal deaths, making it difficult to compute case fatality ratios. Due to the differences in definition and terminology used, as well as nature of data collected, it is difficult to make comparison or reliably interpret data on maternal near miss and case fatality ratios from specific maternal complications (across the studies).Severe acute maternal morbidity also known as "maternal near miss", is defined as "a very ill pregnant or recently woman who would have died had it not been that luck and good care was on her side" . Our revFrom our attempt to harmonize the definition used in the few studies available, the incidence or prevalence ratio of maternal near misses) ranges from 1.1%-10.1% and case fatality ratio from 3.1%-37.4%, indicating wide variation in reported magnitude of the problem and case fatality ratios across the different countries. This review provides invaluable insights into the magnitude of maternal near misses, efficiency of maternal health services and quality of obstetric care in sub-Saharan Africa in the period 1995-2010. The review shows that obstructed labor, obstetric hemorrhage, ruptured uterus and sepsis (in descending order) are the commonest causes of maternal near misses.Our review shows that in the 15 years 1995-2010), there was no standard definition of a maternal near miss. Therefore, different researchers used different definitions or a combination of different definitions of maternal near miss, namely the clinical/diagnostic criterion, the management-based criteria and the criteria of organ/system dysfunction. The advantage of the clinical approach in defining maternal near miss is that it is easy to interpret, appeals to both clinicians and non-clinicians, identified diagnoses tend to mirror the main causes of maternal death and this data is often routinely collected in medical records or hospital registers ,8. The l95-2010, The differences in case fatality ratios are a reliable indicator of quality of care. For instance, in the prospective community-based cohort study by Prual et al  conducteOur review highlights the paucity of nationally or regionally representative studies on maternal mortality or maternal near misses, absence of a uniform definition of maternal near miss events and absence of uniformity in the terminology used to refer to women who have severe obstetric complications. The findings are in agreement with reports of inconsistencies in the way maternal deaths and severe maternal morbidity are classified or reported, as well as absence of standard definitions and criteria for identifying maternal near miss cases ,7,36. SeOur review fulfils requirements in the consensus statement for reporting of systematic reviews of observational studies  in preseBetween sub-Saharan African countries, large differences exist on the magnitude of severe maternal morbidity  in studies reported between 1996 and 2010. This could be due to different contexts/settings and variation in the criteria used to define the near miss morbidity, or rigor used carrying out the study. Researchers should adopt the new maternal death classification system to enable reliable comparisons to be made within and between countries and regions, as well as to enable identification of health system shortfalls that countries need to address.The authors declare that they have no competing interests.DKK conceptualized the study. DKK and OK designed the study protocol and designed the systematic review instruments. DKK, MOO and OK reviewed the literature. DKK wrote the first draft of the manuscript. DKK, MOO and OK contributed to revision of the subsequent draft manuscripts and approved the final version of manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2393/11/65/prepub


Administration of pharmacological doses of catecholamine or adrenergic receptor agonists often results in arrhythmias onset. Results from the present study indicate that stimulation of cardiac adrenergic receptors elicits an extrusion of cellular Mg2+ into the extracellular space. This effect occurs in both perfused hearts and isolated cells within 5-6 min following either β- or α1-adrenergic receptor stimulation, and is prevented by specific adrenergic receptors antagonists. Sequential stimulation of the two classes of adrenergic receptor results in a larger mobilization of cellular Mg2+ provided that the two agonists are administered together or within 1-2 min from each other. A longer delay in administering the second stimulus results in the abolishment of Mg2+ extrusion. Hence, these data suggest that the stimulation of β- and α1-adrenergic receptors mobilizes Mg2+ from two distinct cellular pools, and that Mg2+ loss from either pool triggers a Mg2+ redistribution within the cardiac myocyte. At the sarcolemmal level, Mg2+ extrusion occurs through a Na+/Mg2+ exchange mechanism phosphorylated by cAMP. Administration of quinidine, a patent anti-arrhythmic agent, blocks Na+ transport in a non-specific manner and prevents Mg2+ extrusion. Taken together, these data indicate that catecholamine administration induces dynamic changes in total and compartmentalized Mg2+ pools within the cardiac myocytes, and suggest that prevention of Mg2+ extrusion and redistribution may be an integral component of the effectiveness of quinidine and possibly other cardiac anti-arrhythmic agents. Confirmation of this possibility by future experimental and clinical studies might result in new patents of these compounds as Mg2+ preserving agents.Magnesium (Mg Because αlar Ca2+ , hearts nhibitor , while m1). Table 1 and also  show that pretreatment of cardiac myocytes with KB-R7943, a fairly specific inhibitor of the Na+/Ca2+ exchanger in reverse mode [2+ extrusion elicited by isoproterenol or phenylephine stimulation. Results qualitatively to those reported in perfused hearts were observed in collagenase-dispersed cardiac myocytes incubated under similar experimental conditions  suggest the presence of two distinct Mg2+ cellular pools targeted by α1-and β-adrenergic receptor, respectively. Thus, we assessed whether these two pools could be sequentially mobilized in the same heart by phenylphrine and isoproterenol administered in varying sequence order and with a varying time interval between stimulations. Fig. 5 shows the results of heart’s double perfusion with the two adrenergic agonists administered at 10 min interval from each other. To facilitate comparison, the net amount of Mg2+ extruded by each agonist is reported in . As we can observe, administration of phenylephrine as the first agent does not affect the subsequent release of Mg2+ induced by isoproterenol. Administration of isoproterenol as the first agent, however, decreases the amplitude of phenylephrine-induced Mg2+ extrusion by approximately 40% . Administration of epinephrine as the first agent, instead, decreases the amplitude of phenylephrine-induced Mg2+ extrusion by 88%, and that induced by isoproterenol by approximately 50% (Table 2). Lastly, we varied the interval between administrations of β- and α1-adrenergic agonist. The results reported in (Table 2) indicate that shortening the interval between the two stimulations resulted in a larger phenylephrine-induced Mg2+ extrusion whereas extending the interval resulted in a smaller Mg2+ extrusion. Administration of amiloride at the time of isoproterenol addition abolished the Mg2+ extrusion elicited by β-adrenergic receptor stimulation, and enlarged the subsequent extrusion elicited by phenylephrine (Table 2). When the sequence of agonist was reverse, amiloride did not abolished the effect of phenylephrine, as already observed in , but markedly inhibited isoproterenol effect. Administration of quinidine, instead, resulted in a partial inhibition of both α1- and β-adrenergic receptor stimulations, as reported previously (Table 2). The data reported in  to mobilize Mg2+ from cytoplasm, mitochondria and non-mitochondria compartments. We have already used this approach in other cells with satisfactory results [2+-mobilizing agents is reported as an onset in . Fig. 6 also reports an estimate of the absolute content of Mg2+ mobilized by digitonin, uncoupler (FCCP) and A23187 ionophore from unstimulated cardiac myocytes and from cells previously treated for 8 min with phenylephrine, isoproterenol, and epinephrine. As the results indicate, phenylephrine stimulation largely mobilized Mg2+ from non-mitochondrial compartments (largely the sarcoplamisc reticulum) while isoproterenol stimulation depleted Mg2+ from cytoplasm and mitochondria. Consistent with its role of mix agonist, epinephrine mobilized Mg2+ from all compartments. Similar protocol was also applied to cardiac myocytes previously stimulated with phenylephrine or isoproterenol for 6 min. The results reported in  are suggestive of a Mg2+ redistribution among compartments based upon the type and duration of the stimulus applied.Because we lack advance techniques to detect in situ changes in cellular Mg results . A typic2+ extrusion from various tissues including cardiac myocytes [1-adrenergic receptor, the effect depending on the receptor representation in the tissue, the catecholamine of choice (i.e. norepinephrine or epinephrine), and the experimental conditions. For example, studies carried out in our laboratory indicate that stimulation of 1-adrenergic receptors induce Mg2+ extrusion in liver cells, where the process is associated with glucose output [Elevated or persistent adrenergic stimulation of cardiac cells has been associated with increased incidence of cardiac arrhythmias. Adrenergic stimulation has been reported to induce Mgmyocytes -11. Becamyocytes -11. β the output ,24.1-adrenergic receptorβ- and stimulation on Mg2+ extrusion and cellular Mg2+ redistribution among compartment as well as the effect of the anti-arrhythmic drug quinidine, which has been reported to inhibit Mg2+ extrusion through the Na+/Mg2+ exchanger [In the present study we attempted to address two distinct issues. The relative effect of xchanger .1- and β- adrenergic receptors results in two well distinct Mg2+ extrusion processes when stimulatory agents for each receptor class are administered individually. The administration of catecholamine, which stimulate both classes of adrenergic receptors with different specificity, results in a Mg2+ extrusion equivalent to the sum of the amounts released by phenylephrine and isoproterenol individually. This is consistent with the notion that phenylephrine and isoproterenol target two distinct cellular Mg2+ pools within the cardiac myocytes. This observation is further congruent with the notion that β-adrenergic receptor stimulation mobilizes Mg2+ predominantly from cytoplasm and mitochondria whereas α1-adrenergic receptor stimulation mobilizes Mg2+ predominantly from the reticular pool . Interestingly, the operation of this extrusion mechanism is independent of cAMP-mediated phosphorylation, and can be related albeit indirectly to the activation of the α1-adrenergic receptor and associated Ca2+-mediated signaling [2+ entry across the cell membrane. This aspect will be consistent with the reduced phenylephrine-induced Mg2+ extrusion that occurs in cardiac cells in the presence of nifedipine, a L-type channel inhibitor commonly used under experimental and clinical conditions. Irrespective of the mechanism involved, it appears evident that quinidine and nifedipine, and perhaps other agents of similar classes, block Mg2+ extrusion and also cellular Mg2+ redistribution, thus affecting the level of Mg2+ retained within organelles and associated metabolic functions. In this contest, it would be important to assess in future studies to which extent prevention of Mg2+ extrusion and redistribution by anti-arrhythmic drugs and Ca2+ channel blockers is integral part of the pharmacological effectiveness of these therapeutic agents. An interesting observation generated from our studies is that the anti-arrhythmic drug quinidine inhibits by approximately 40-45% the Mgspecific ,23. Our channels . In turnmyocytes  and isolmyocytes  indicateignaling ,22, incl2+ from cardiac ventricular myocytes, which results in dynamic changes in total and compartmentalized Mg2+ pools within the myocytes. In addition, our data suggest that prevention of Mg2+ extrusion and redistribution may represent an integral component of the effectiveness of at least some anti-arrhythmic drugs and Ca2+-channel blocking agents. Additional studies both at the experimental and the clinical levels are necessary to confirm our initial observation and its pharmacological relevance. If confirmed, this observation might result in patenting (some of) the existing anty-arrhythmic and L-type-Ca2+ channel blockers as Mg2+ preserving agents. Additionally, it might lead to the development of new therapeutic agents that are more effective at preventing cellular Mg2+ loss, with consequent enhanced effectiveness and reduced toxicity or intrinsic arrhythmogenicity, which often limit their pharmacologic use. Our results indicate that catecholamine administration induces a major efflux of Mg


HNF1A) transcription factor, (2) activation of the WNT/β-catenin by CTNNB1 mutations, or (3) activation of the IL6/STAT3 pathway by somatic mutation of IL6ST, GNAS, or STAT3. Here, we will review the different molecular classes of HCA.Hepatocellular adenomas (HCAs) are benign tumors developed in normal liver most frequently in women before menopause. HCAs lead to diagnostic pitfalls and several difficulties to assess the risk of malignant transformation in these young patients. Recent advances in basic knowledge have revealed a molecular classification related to risk factors, pathological features, and risk of transformation in hepatocellular carcinoma. Three major molecular pathways have been identified altered in specific HCA subgroups that are defined by either (1) inactivation of hepatocyte nuclear factor 1A ( Hepatocellular tumors deriving from monoclonal proliferation of hepatocytes are classically divided in benign hepatocellular adenoma (HCA) and malignant hepatocellular carcinoma (HCC). HCAs are rare tumors most frequently-developed in women before menoaupose and after a long-term use of oral contraception . Other rHNF1A, as the first driver gene inactivated by mutation in hepatocellular adenomas . Th. Th34]. HNF1A in HCA, we also showed that HNF1A somatic mutations were frequently caused by G to T transversion suggesting a genotoxic exposure at the origin of the mutations [In the canonical point of view, malignant hepatocellular tumors (HCC) arise on chronic liver disease, mainly cirrhosis or chronic HBV infection, whereas hepatocellular benign tumors are developed on normal liver. However, several clinical, pathological, and molecular observations have challenged these dogmas. First, HCC could develop on normal liver, and predisposing genetic factor and genetic drivers involved in tumor initiation remain poorly described . A simplutations . Causes HNF1A germline mutation related to MODY3 diabetes and GNAS mosaic somatic mutations related to McCune Albright disease are strong risk factors of adenomas occurrence [HNF1A germline mutation, GNAS mosaic postzygotic mutation, and others unknown modifier genes) and environmental factors (oestrogen and androgen expositions) [CYP1B1-inactivating mutation in 12.5% of patients developing HNF1A-inactivated HCA. Moreover, when analyzing the spectrum of somatic mutations in HNF1A, we identified a genotoxic signature typical of molecule inducing adduct to DNA at guanine [In this line, predisposing genetic factors like currence , 50. Morcurrence –63. All sitions) , 60, 64.sitions) . We idenβ-catenin, other genetic alterations leading to full malignant transformation have to be deciphered. Finally, several driver genes of benign tumorigenesis are still unknown, especially in the group of inflammatory HCA without known driver mutations and unclassified HCA. Ultimately, these genetic alterations will constitute therapeutic target for biotherapy that will be used in unresectable HCA or in other malignancies harboring the same genetic events.A long path has been walked in the area of hepatocellular benign tumors since Edmonson described the association between HCA and oral contraception . Now, th


Cognitive factors associated with drinking behavior such as positive alcohol expectancies, self-efficacy, perception of impaired control over drinking and perception of drinking problems are considered to have a significant influence on treatment effects and outcome in alcohol-dependent patients. However, the development of a rating scale on lack of perception or denial of drinking problems and impaired control over drinking has not been substantial, even though these are important factors in patients under abstinence-oriented treatment as well as participants in self-help groups such as Alcoholics Anonymous (AA). The Drinking-Related Cognitions Scale (DRCS) is a new self-reported rating scale developed to briefly measure cognitive factors associated with drinking behavior in alcohol-dependent patients under abstinence-oriented treatment, including positive alcohol expectancies, abstinence self-efficacy, perception of impaired control over drinking, and perception of drinking problems. Here, we conducted a prospective cohort study to explore the predictive validity of DRCS.Participants in this study were 175 middle-aged and elderly Japanese male patients who met the DSM-IV Diagnostic Criteria for Alcohol Dependence. DRCS scores were recorded before and after the inpatient abstinence-oriented treatment program, and treatment outcome was evaluated one year after discharge.Of the 175 participants, 30 were not available for follow-up; thus the number of subjects for analysis in this study was 145. When the total DRCS score and subscale scores were compared before and after inpatient treatment, a significant increase was seen for both scores. Both the total DRCS score and each subscale score were significantly related to total abstinence, percentage of abstinent days, and the first drinking occasion during the one-year post-treatment period. Therefore, good treatment outcome was significantly predicted by low positive alcohol expectancies, high abstinence self-efficacy, high perception level of impaired control over drinking, and high perception level of drinking problems measured by DRCS.The DRCS was considered to have satisfactory predictive validity, which further supports our previous findings. It was suggested that DRCS is a promising rating scale for evaluating multidimensional cognitive factors associated with drinking behavior in alcohol-dependent patients under abstinence-oriented treatment. There are various cognitive factors associated with drinking behavior which influence treatment outcome in alcohol-dependent patients -6. For eOther important cognitive factors associated with drinking behavior upon determining whether or not alcohol-dependent patients will drink alcohol include outcome expectancies and self-efficacy. Alcohol outcome expectancies include positive expectancies anticipating positive consequences of drinking (such as “I expect to be the life and soul of the party if I have a few drinks”) and negative expectancies anticipating negative consequences of drinking (such as “I expect to have a hangover if I have a few drinks”) . In geneAs described above, cognitive factors associated with drinking behavior in alcohol-dependent patients extend over a wide range and include perception of drinking problems, perception of impaired control over drinking, readiness to change, motivation, alcohol outcome expectancies, and self-efficacy. These cognitive factors are considered to play an important role as predictors of treatment outcome. Therefore, various rating scales have been developed in order to measure cognitive factors associated with drinking behavior  with factor analysis. All items have a factor load of 0.40 or greater. The cumulative contribution rate of the three factors was 58.2%  for three to four weeks at the Internal Medicine Ward, and were transferred to the Alcohol Treatment Ward when they were at a mentally and physically stable stage. After the transfer, the detailed purpose of this study was explained to the participants verbally and in writing, and written consent was obtained from each participant for conducting DRCS as well as cooperating with the outcome survey after discharge.First, participant characteristics  were assessed prior to implementation of the inpatient treatment program, and DRCS was conducted. After the nine-week inpatient treatment program was completed including assessment and feedback regarding alcohol problems in patients, education on alcohol dependence, motivational approaches to abstinence, coping-skills training, facilitation to participate in a self-help group, etc., DRCS and SDS were conducted again (this inpatient treatment is an abstinence-oriented program). Finally, one year after discharge, an investigation was conducted by face-to-face interview or by phone with patients, families, and municipal welfare office staff. The treatment outcome at the one-year follow-up was assessed by assigning (a) patients with “0” drinking days as the “abstinence group” and (b) patients with “1 or more” drinking days as the “drinking group.”DRCS was developed to evaluate cognitive factors associated with drinking behavior in alcohol-dependent patients who received abstinence-oriented treatment; therefore, the treatment goal for these patients is total abstinence. However, in order to increase the predictive validity of DRCS, not only total abstinence but also two other treatment outcomes associated with abstinence (the percentage of abstinent days during the one-year post-treatment period and the number of days until the first drinking occasion after discharge) were added as criteria to evaluate the predictive validity. Thus, the predictive validity of DRCS was evaluated by three treatment outcomes during the one-year post-treatment period: (1) total abstinence, (2) percentage of abstinent days and (3) the first drinking occasion.t-test. Next, DRCS scores before and after the treatment program were compared by paired t-test.First, the treatment outcome at the one-year follow-up was compared in terms of participant characteristics as well as DRCS scores by chi-square test or unpaired Furthermore, binominal logistic regression analysis was conducted using the forced entry method, with total abstinence as a dependent variable and the total post-treatment DRCS scores, subscale scores, changes in each DRCS score from baseline to end of treatment and age as well as participant characteristics showing significant difference by univariable analysis as independent variables, in order to investigate how the cognitive factors influence treatment outcome under conditions excluding the influence of confounding factors such as participant characteristics.The extent to which the total post-treatment DRCS scores and subscale scores influence the percentage of abstinent days during the one-year post-treatment period was also investigated. For this purpose, the generalized estimating equation (GEE) analysis was used with the percentage of abstinent days as the dependent variable and the total post-treatment DRCS scores and subscale scores, age, and participant characteristics indicating significant differences in univariable analysis as independent variables.Finally, the influence of the total post-treatment DRCS scores and subscale scores was examined in terms of the number of days until the first drinking occasion after discharge. Proportional hazard analysis was conducted using the forced entry method, with the first drinking occasion as the dependent variable and the total post-treatment DRCS scores and subscale scores, age, and participant characteristics indicating significant differences in univariable analysis as independent variables.SPSS 16.0 software was used for statistical analysis.Of the 175 participants in this study, 20 requested discharge during the course of the inpatient treatment program, and 10 were unavailable for follow-up one year after discharge. Thus, the number of subjects for analysis in this study is 145. Among the 145 participants whose answers were successfully obtained in the outcome survey, 53 answers were from face-to-face interviews, 40 from phone interviews, 11 from phone interviews with participants and families, 36 from phone interviews with families only, and five from phone interviews with municipal welfare office staff only.t-test = 1.79, df = 173, p = 0.076; mean “expectancy and resignation” score: 28.1  vs. 27.6 , unpaired t-test = 0.40, df = 173, p = 0.689; mean “perception of impaired control” score: 22.0  vs. 21.7 , unpaired t-test = 0.27, df = 47, p = 0.787; mean “perception of drinking problems” score: 18.2  vs. 19.0 , unpaired t-test = −0.80, df = 173, p = 0.425; mean “total” score: 68.3  vs. 68.3 , unpaired t-test = 0.01, df = 51, p = 0.992).The demographic characteristics of the participants are shown in Table t-test = −6.53, df = 144, p < 0.001; “perception of impaired control” increased from 22.0  to 25.0 , paired t-test = −6.10, df = 144, p < 0.001; “perception of drinking problems” increased from 18.2  to 20.8 , paired t-test = −8.15, df = 144, p < 0.001; and “total” increased from 68.3  to 77.1 , paired t-test = −8.97, df = 144, p < 0.001).When total DRCS scores and subscale scores were compared before and after treatment, a significant increase was seen for both scores . Characteristics showing a significant difference between the abstinence group and the drinking group included the number of admissions to a psychiatry department to treat alcohol dependence, history of finger tremor due to alcohol withdrawal, employment status, educational history, and history of illegal drug usage or abuse. A significant difference was not seen between the abstinence group and the drinking group regarding the pre-treatment DRCS scores; however, both the total post-treatment DRCS scores and subscale scores were significantly higher in the abstinence group than in the drinking group.The relationship between participant characteristics and total abstinence at the one-year follow-up in the univariable analysis is shown in Table The adjusted odds ratios for total abstinence during the investigation period are shown in Table Both the total DRCS score and each subscale score were significantly related to total abstinence during the one-year investigation period. The adjusted odds ratio (AOR) for total abstinence by one-point increase of each DRCS score was 1.12 for “expectancy and resignation,” 1.10 for “perception of impaired control,” 1.12 for “perception of drinking problems,” and 1.05 for “total” score.However, the odds ratio for changes in each DRCS score adjusted under the same conditions was not significant in terms of total abstinence .The average percentage of abstinent days during the one-year post-treatment period was 45.4%  for 92 patients in the drinking group. The total post-treatment DRCS score and subscale scores were significantly related to the percentage of abstinent days in the GEE analysis with the percentage of abstinent days as the dependent variable  for 92 patients in the drinking group. Both the total DRCS score and each subscale score were significantly related to the first drinking occasion .Click here for fileFactor analysis of DRCS .Click here for file


Genetic analysis of factors affecting risk to develop excessive ethanol drinking has been extensively studied in humans and animal models for over 20 years. However, little progress has been made in determining molecular mechanisms underlying environmental or non-genetic events contributing to variation in ethanol drinking. Here, we identify persistent and substantial variation in ethanol drinking behavior within an inbred mouse strain and utilize this model to identify gene networks influencing such “non-genetic” variation in ethanol intake. C57BL/6NCrl mice showed persistent inter-individual variation of ethanol intake in a two-bottle choice paradigm over a three-week period, ranging from less than 1 g/kg to over 14 g/kg ethanol in an 18 h interval. Differences in sweet or bitter taste susceptibility or litter effects did not appreciably correlate with ethanol intake variation. Whole genome microarray expression analysis in nucleus accumbens, prefrontal cortex and ventral midbrain region of individual animals identified gene expression patterns correlated with ethanol intake. Results included several gene networks previously implicated in ethanol behaviors, such as glutamate signaling, BDNF and genes involved in synaptic vesicle function. Additionally, genes functioning in epigenetic chromatin or DNA modifications such as acetylation and/or methylation also had expression patterns correlated with ethanol intake. In verification for the significance of the expression findings, we found that a histone deacetylase inhibitor, trichostatin A, caused an increase in 2-bottle ethanol intake. Our results thus implicate specific brain regional gene networks, including chromatin modification factors, as potentially important mechanisms underlying individual variation in ethanol intake. Over 121 million Americans drink alcohol, while less than 10% of the population drinks excessively Despite such progress on identifying genetic influences in alcoholism, little progress at the molecular level has revealed mechanisms that mediate environmental influences on ethanol behaviors or alcoholism. It is well documented that environmental influences such as stress or exposure to conditional stimuli can modify ethanol drinking or cause recidivism in abstinent alcoholics. Understanding the molecular mechanisms underlying such environmental influences on ethanol behaviors would augment the genetic progress mentioned above.C57BL/6 (B6) inbred mice have been widely used as a model for studying alcohol abuse related behaviors and the genetic basis of alcohol abuse since these mice voluntarily consume large volumes of unadulterated ethanol Here, we demonstrate a remarkable degree of individual variation in ethanol drinking behavior across individual mice from the C57BL/6NCrl inbred line. We have performed whole genome expression profiling in individual mice to finely dissect molecular factors underlying this individual variation in ethanol drinking behavior. We hypothesized that an as yet unidentified non-genetic factor has caused long-lasting brain signaling alterations that influence ethanol preference and intake in these mice. By characterizing gene networks differentially expressed between ethanol preferring and avoiding mice, we have identified putative epigenetic mechanisms such as alterations in chromatin acetylation that may regulate gene transcription and influence drinking patterns. We expect that these studies may contribute to the identification of novel targets for pharmacotherapy in alcoholism.All procedures were approved by Virginia Commonwealth University Institutional Animal Care and Use Committee under protocol numbers AM10332 and AM10139, and followed the NIH Guide for the Care and Use of Laboratory Animals .Male C57BL/6NCrl mice (age 42–49 days) from Charles River Laboratories  were habituated to the vivarium (5 mice/cage) for 1 week followed by individual housing for 1 week prior to beginning drinking experiments. Cages and bedding  were changed weekly. Mice were housed in a temperature and light controlled room  with free access to standard chow  and water.Experiment 1: Voluntary two-bottle choice drinking was performed as described previously Experiment 3: For studies on ethanol preference in littermates, two cohorts of male B6 littermates were purchased as littermates from Charles River. Breeding at Charles River facility used two females paired to one male. Offspring were weaned at day 21, shipped to our vivarium at day 35 and remained housed as littermates until the beginning of the study. 10 litters were represented with 3–5 males per litter . Mice were individually housed for 7 days then presented with 10%(w/v) ethanol for 18 h/day in a two-bottle choice paradigm for 14 days.Experiment 4: Taste preference for a bitter or sweet solution was measured using quinine or saccharin vs. water in a two-bottle choice paradigm. Mice (n = 16) were tested for ethanol preference for 14 days as in experiment 1 but without deprivation periods, and then allowed to rest for 7 days with only water and food available. Half the mice were given two bottles containing either a 0.1 mM quinine solution or tap water and the others given a choice between 0.033% saccharin/water. Bottles were alternated every other day. Consumption of quinine/water or saccharin/water was measured daily for 3 days (18 h/day) after which the other tastant was offered for 3 days in a counterbalanced design.Experiment 5: 18 male C57BL/6NCrl mice, acclimated to a reverse-light cycle, were used to test effects of trichostatin A  on ethanol drinking. Voluntary ethanol drinking was initiated as before except these mice had access 24 hours/day, changed at 1200 h. Following 7 days baseline ethanol access, mice were divided into two groups balanced for baseline intake. Mice in the TSA group (n = 9) were injected with 2 mg/kg TSA i.p.  for 5 days. Control mice (n = 9) were injected with vehicle. Mice had continuous ethanol access during the entire study .Experiment 6: Western blotting for acetyl-Histone-H4 was used to verify CNS activity of i.p. TSA. C57BL/6NCrl mice (n = 12) received a single i.p. injection of 2 mg/kg TSA or vehicle. After 24 hours, nucleus accumbens was dissected for Western blotting as described below.The effect of TSA on ethanol metabolism was assayed in a separate group of mice (Experiment 7). Male C57BL/6NCrl mice  received 2 mg/kg TSA or vehicle for 5 consecutive days. On day 6, all mice were injected with 2 mg/kg ethanol. Trunk blood was harvested from mice at 10, 60, 120 and 180 minutes following ethanol administration. Whole blood samples (20 µl) were placed into 20-ml headspace vials with 960 µl water and 20 µl 1-propanol internal standard. Samples were tested for ethanol concentration using a Hewlett Packard 5890A gas chromatograph equipped with a flame ionization detector, 2 meter 5% Carbowax 20M 80/120 mesh packed column  and CTC Combi-Pal headspace autosampler. Data were acquired by Clarity software  and analyzed by linear regression with no weighting. A 7-point calibration curve preceded the analysis and quality control ethanol standards were interspersed with each set of samples. Up to 3 replicates were analyzed from each animal and averaged if sufficient blood was collected.µg) was transcribed into double-stranded cDNA using the One-cycle Targeting and Control Reagent kit . Biotin-labeled cRNA was synthesized from cDNA, purified and fragmented according to manufacturer's instructions.Total RNA was extracted from PFC, NAc and VMB from individual mice in STAT 60 reagent  according to the manufacturer's protocol. RNA concentration and quality was assessed by Experion automated electrophoresis . Total RNA  was hybridized to a single microarray for each brain region . Samples were analyzed on oligonucleotide arrays (Mouse Genome 430A 2.0 array) containing >22,000 well-characterized genes and expressed sequence tags. Array hybridization and scanning were performed exactly according to manufacturer's protocols (Affymetrix).Microarray data were processed using GeneChip Operating Software v4.1  and normalized to a mean total hybridization intensity of 190. All raw microarray data is deposited at the Gene Expression Omnibus repository (GEO) under accession number GSE26506. All microarray data is MIAME compliant. Array quality was assessed by accepting arrays with scaling factor <3, 3′–5′-actin ratio <2, and by examining linearity and inter-chip correlations of intensity values. Arrays determined to be acceptable (57/58) were further analyzed using the Robust Multichip Average (RMA) low-level analysis algorithm to summarize probeset expression data www.ingenuity.com) and Bibliosphere (http://www.genomatix.de). These tools utilize biomedical literature associations to annotate genes with biological functions and cellular components. IPA also generates networks of interrelated genes based on their curated knowledge base.The Expression Analysis Systematic Explorer (EASE version 1.21) As an alternative to correlating gene expression to the average of the last 8 days of ethanol drinking data, we conducted a principal components analysis to reduce the number of covariates and identify the few principal components that account for a sufficient proportion of the variability and contain almost as much information as the full data set. The first 2 principal components accounted for 0.77 of the total variance. For each brain region, probeset-specific linear models predicting expression as a function of the two independent principal components were fit. An overall F-test was used for calculating P-values for each probe set level linear model. Genes significant at p<0.05 level were selected for further bioinformatics analysis and comparison with results from using average ethanol intake over the last eight days.d|≥0.5 and q<0.05 Genes significantly correlated to ethanol drinking patterns in the present study, using a false discovery rate of 1%, were analyzed for overlap with previously published gene sets having expression significantly different between alcohol-preferring or non-preferring mouse models based on the criteria |Twenty-one C57BL/6NCrl mice (Exp. 2) voluntarily consumed ethanol as described above and brains were harvested six days following the last drinking session. Nucleus accumbens was homogenized in NP40 buffer with protease inhibitors . Western blotting was performed as described C57BL/6 (B6) male mice from Charles River Laboratories consumed a substantial amount of ethanol, 6.47±0.99 g/kg/18 h, in the voluntary two-bottle choice (10% w/v ethanol or water) self-administration paradigm. Interestingly, mice showed a large degree of inter-individual variation in ethanol drinking , rangingTo assess whether taste discrimination was contributing to variation in ethanol drinking, we performed an additional experiment (Exp. 4) where mice (n = 16) were assessed for ethanol drinking, followed by studies on preference for quinine (0.1 mM) or saccharin (0.033%). While there was some individual variation in quinine consumption, preference for saccharin  or quinine  showed no significant correlation to ethanol preference (data not shown). These results argue against sweet or bitter taste as a major contributing factor for the observed individual differences in ethanol preference.Additionally, we determined whether the observed individual variation in ethanol drinking was due to robust differences between litters (Exp. 3). In two separate cohorts consisting of 10 litters , ethanol intake over 14 days of baseline drinking did not differ between litters  = 1.258 p = 0.2967, We hypothesized that persistent individual variation in ethanol drinking behaviors within an inbred strain might be caused by differential basal gene expression patterns generated by unknown environmental influences. Further, such differential gene expression patterns could be used to identify molecular pathways contributing to individual variation in ethanol drinking. We profiled 3 brain regions in individual mice: nucleus accumbens (NAc), prefrontal cortex (PFC) and ventral midbrain region (VMB). These brain regions were chosen because they are major components of the mesocorticolimbic dopamine reward pathway activated by ethanol and other drugs of abuse To identify molecular factors related to ethanol drinking behaviors, gene expression patterns were first correlated to a drinking template created from the last 8 days of ethanol access following a third round of ethanol deprivation see . This deTo identify gene expression correlated with drinking behavior, we also performed a principal component analysis on the daily drinking activity data to reduce the number of covariates, rather than averaging the drinking data over an interval. The first two principal components (PC) accounted for 77% of the total variance. For each brain region, probe set-specific linear models predicting expression as a function of the two independent principal components were fit. The number of transcripts which fit the linear model at a level p<0.05 was 547 in NAc, 670 in PFC and 725 in VMB . When thGene lists from microarray analyses were analyzed for over-representation of biological functions or gene network relationships using several different tools as described in The 889 transcripts from NAc correlating with average drinking values were analyzed by EASE Hdac11 (NM_144919.2), Rbbp4 (NM_009030.3), Rbbp7 (NM_009031.3), Sap18 (NM_009119.3) and Suds3 (NM_178622.4). All genes in this network were significantly correlated to average ethanol drinking  are involved in DNA methylation events and work concurrently to repress transcription Biological functions of our gene lists were further investigated using the curated knowledge base in Ingenuity Pathway Analysis. This tool generates networks of genes with known interactions or biological function. One of the top networks generated through this process is shown in drinking . Rbbp4 aMyst3 (NM_001081149), Hdac11 and Ehmt2 (NM_147151) were significantly different in high versus low drinkers by t-test at p<0.05. Myst3, Myst histone acetyl transferase 3, is a member of a mouse histone acetyl transferase (HAT) complex that increases DNA transcription Rbbp4, Rbbp7), had intrinsic HDAC activity (Hdac11) or were involved in methylation of DNA (Men1 (NM_001168488), Mbd2 (NM_010773.2), Mll1 (NM_001081049), Ehmt2). These genes are believed to cause transcriptional silencing by removing acetyl groups from chromatin or increasing DNA methylation.The relative expression of select genes in this network in the top quartile of high ethanol drinkers (>7 g/kg/18 h) and the bottom quartile of low ethanol drinkers (<2 g/kg/18 h) are summarized in Ap2a1 (NM_025606), Ap2a2 (NM_007459), Ap2m1 (NM_207255), Dnm1 (NM_144516), Dnm1l (NM_028661), Vamp3 (NM_009498), and Vamp4 (NM_016796.3)) and synaptic vesicle biogenesis (Sh3gl2 (NM_019535), Sh3glb1 (NM_175141)) were generally positively correlated to ethanol intake. This suggests that synaptic vesicle recycling may be increased in mice prone to drinking greater amounts of ethanol. Conversely, low drinking mice had higher Bdnf (NM_001037955) expression. Bdnf may play a role to increase synaptogenesis in these studies as it has been implicated in plasticity from multiple drugs of abuse Further network analysis identified genes involved in synaptic vesicle formation and recycling see . Genes i+K+ ATPase activity and protein kinase activity , Grin3b (NM_028388)) and the kainite receptor (Grik1 (NM_010348)), as well as genes that bind (Htt (NM_010414)) or are regulated by glutamate receptors (Dlg4 (NM_001109752) aka Psd95). The NR2b subunit of the NMDA receptor was positively correlated to ethanol drinking with the lowest drinking mice having lower expression (Grik1) were correlated negatively to ethanol drinking. Tyrosine hydroxylase, the rate-limiting enzyme in catecholamine synthesis involved in the conversion of tyrosine to dopamine, was also positively correlated to ethanol drinking  was chosen for its documented role in synaptic vesicle trafficking. In a separate cohort of mice (Exp. 2), RAB3A expression was determined in high  and low  drinking mice . RAB3A ebasal brain gene expression across multiple mouse strains with divergent ethanol drinking phenotypes. A large meta-analysis of this data identified gene expression correlated with ethanol drinking behavior across these multiple genetic models −34, Chi-square analysis) were also identified in the meta-analysis (see Stxbp1 (NM_009295)) has been previously identified as a putative candidate for an ethanol-drinking locus on Chromosome 2 −18, Chi-square) were also identified by the meta-analysis  were in common with the meta-analysis results  did not allow a definitive conclusion as to whether TSA affected the overall distribution of drinking values, since most animals showed increased drinking following TSA (not shown).et al. reported that within-strain preference variation was not correlated with gender or ethanol metabolism, and could not be altered by simple environmental disturbances Our studies here showed that C57BL/6NCrl mice express a striking degree of stable, inter-individual variation in ethanol drinking behavior with greater than 10-fold differences within a drinking session. We suspect these differences were generated by subtle environmental differences such as rearing behaviors The studies here employed a unique experimental design that allowed long-term measures of ethanol drinking behavior, ensured that such behavior was stable upon reinstatement, and permitted assaying gene expression differences in individual animals off ethanol. This allowed identification of expression patterns presumably “predictive” of drinking behavior rather than simply resulting from such. However, even with the current design, we cannot totally eliminate the possibility that some of our gene expression results reflect, rather than cause, individual variation in drinking behavior. Additionally, even though the behavioral data showed extinction of an ethanol deprivation effect after several cycles of withdrawal and reinstatement , we cannThe current studies showed a potential role for epigenetic regulation of ethanol preference in B6 mice. In the NAc, genes with chromatin remodeling Gene Ontology function or classified in the HDAC complex had differential expression between high and low ethanol-drinking animals see . IntriguBdnf promoters, leading to decreased Bdnf transcription Inhibiting HDAC activity with TSA injections increased ethanol intake above baseline and vehicle-treated levels, supporting a role for chromatin modifications in the modulation of ethanol preference. The complex changes in chromatin modification gene expression made it difficult to predict how directly altering histone acetylation might affect drinking behavior. We suggest, however, that any TSA-induced change in ethanol drinking across individuals or as a population is supportive of our hypothesis regarding a role of chromatin modification in driving individual variation in ethanol intake. This data is the first to show modulation of drinking behavior by altering chromatin acetylation. However, evidence of ethanol-induced chromatin remodeling has been reported in hepatocytes Stxbp1 was previously identified as a candidate gene for a mouse Chr2 ethanol preference locus C. elegans and mice Our bioinformatics analysis of gene expression correlating with ethanol drinking also identified gene networks involved in synaptic vesicle biogenesis and recycling . Many ofBdnf mRNA levels and individual ethanol consumption. BDNF regulates multiple synaptic vesicle-related proteins, including several listed in Bdnf under-expression in Bdnf+/- mice caused increased ethanol consumption, consistent with Bdnf mRNA expression observed in the current study, where Bdnf is lowest in mice with the highest ethanol intake. We do not believe, therefore, that Bdnf expression levels seen in our studies were secondary to ethanol exposure itself. In support of this, we and other investigators have shown that acute ethanol injection (2 g/kg i.p.) in B6 or D2 mice increases Bdnf expression and that after 4 weeks of 2-bottle choice ethanol drinking, Bdnf is increased in the dorsal striatum versus non-ethanol controls Bdnf expression in the low drinking mice was possibly a causal factor in individual drinking behavior variance, rather than secondary to drinking behavior itself. We cannot currently exclude the possibility that ethanol drinking followed by withdrawal (4 days) caused the correlated changes in Bdnf expression. Together, these findings on Bdnf and synaptic vesicle-related gene expression are strong evidence supporting an important link between regulation of synaptic vesicles and individual variation in ethanol intake.We also identified an inverse correlation between Bdnf expression only being regulated in nucleus accumbens In the present study, many of the most robust gene expression changes were found in the nucleus accumbens and prefrontal cortex. Brain regional differential gene expression is not surprising considering the proposed different roles of each region in ethanol responses Importantly, we found significant overlap between our gene lists and a previously published meta-analysis of basal brain gene expression across mouse strains with differing ethanol preference In conclusion, the current experiments have described persistent inter-individual variation of ethanol drinking behaviors in B6 mice and, more importantly, they define gene expression networks that may underlie these individual differences. This study utilizes variation within an inbred strain to minimize genetic influences, isolating changes in gene expression due specifically to environmental factors. These experiments have identified several gene networks previously implicated in responses to ethanol in the NAc and PFC: glutamate signaling, BDNF and genes involved in synaptic vesicle function. Perhaps most importantly, our expression studies and behavioral analysis following histone deacetylase inhibition implicate epigenetic factors involving chromatin acetylation and/or methylation as contributing to environmental modulation of ethanol intake. Defining specific gene networks targeted by these epigenetic modifications is an important goal of ongoing studies. The novel findings presented here could contribute to understanding mechanisms involved in individual risk for alcohol abuse and alcoholism in humans. Future work will focus on characterizing the genesis and implications of gene network alterations and epigenetic modifications associated with variation in ethanol drinking.Figure S1Average Ethanol Intake Over 16 Days Of Access. Ethanol intake was significantly increased following repeated ethanol deprivations . Ethanol consumption did not differ from baseline after the third deprivation .(TIF)Click here for additional data file.Figure S2Genes Differentially Regulated In Ethanol Drinking Mice.A. Venn diagram overlapping and non-overlapping genes in each brain region significantly correlated to drinking at FDR<0.01. Region-specific expression patterns are represented as shaded circles (nucleus accumbens (NAc), dark; prefrontal cortex (PFC), open; ventral midbrain (VMB), light). B–D. Hierarchical clustering of transcripts significantly correlated to ethanol drinking in the NAc (B), PFC (C) and VMB (D). Genes that overlap with the meta-analysis are labeled in blue. Genes that overlap with the principal component analysis are labeled in orange. Red color indicates higher relative expression and green indicates lower expression. Columns are arranged according to drinking behavior averaged over the last 8 days of intake, with low drinking mice on the left, progressing to higher drinking mice on the right.(TIF)Click here for additional data file.Figure S3Western Blot Analysis Of TSA Effects On Histone H4 Acetylation. Western blotting for acetyl-Histone-H4 was used to verify CNS activity of i.p. TSA. C57BL/6NCrl mice (n = 12) received a single i.p. injection of 2 mg/kg TSA or vehicle. After 24 hours, nucleus accumbens was dissected for Western blotting for acetyl-histone H4 (upper panel) or GAPDH as a loading control (lower panel). Results verify increased H4 acetylation in NAc after TSA treatment.(TIF)Click here for additional data file.Table S1Average Ethanol Intake and Preference for six C57BL/6NCrl litters. Average ethanol intake and preference for six separate litters of male mice (n = 4−5/litter) was calculated over 14 days of 24 h access to 2-bottle choice ethanol. There was no significant difference in intake or preference as reported in (XLSX)Click here for additional data file.Table S2Results from Correlation with Average Drinking. Data are gene lists and statistics for gene expression correlating with individual average ethanol intake from the last 8 days of ethanol access following a third round of ethanol deprivation. Data are presented for each brain region separately.(XLSX)Click here for additional data file.Table S3Gene Lists from Principal Component Analysis. Data are gene lists and statistics for gene expression correlating with ethanol intake data subjected to principal component analysis as in (XLSX)Click here for additional data file.Table S4Over-Represented Gene Categories From Gene Expression Significantly Correlated To Average Ethanol Drinking. Gene data from (XLSX)Click here for additional data file.Table S5Gene Categories Significantly Over-Represented From Principal Component Analysis In Ethanol Drinking C57BL/6NCrl Mice. Gene set data from (XLSX)Click here for additional data file.


The literature shows an inconsistent relationship between lifestyle behaviors and metabolic syndrome (MetS), especially in the elderly. We designed this study to investigate the interrelationships among cigarette smoking, tea drinking and MetS, and to verify the factors associated with MetS in elderly males dwelling in rural community. In July 2010, with a whole community sampling method, 414 male subjects aged over 65 dwelling in Tianliao township were randomly sampled. The response rate was 60.8%. Each subject completed the structured questionnaires including sociodemographic characteristics, habitual behaviors (including cigarette smoking and tea drinking habits) and medical history. After an overnight fast, the laboratory and anthropometric data were obtained. MetS was confirmed according to the criteria defined by the modified NCEP ATP III for the male Chinese population. Subjects were split into either non-MetS or MetS groups for further analysis. Of the 361 subjects with complete data, 132 (36.6%) elderly men were classified as having MetS. Using binary logistic regression, body mass index, serum uric acid, high sensitivity C-reactive protein, HOMA index, current smokers , total smoking amount > = 30  and more than 20 cigarettes daily  were positively associated with MetS. Current un- or partial fermented tea drinker , tea drinking habit for 1–9 years  and more than 240cc daily  were negatively associated with MetS. In conclusion, this study suggests that smoking habit was positively associated with MetS, but tea drinking habit was negatively associated with MetS in elderly men dwelling in rural community. Metabolic syndrome (MetS) has become a major health hazard due to its increasing prevalence worldwide The relationship between lifestyle behaviors and MetS has recently attracted more attention p = 0.17) and age distribution between responders and non-responders (p>0.05). Finally, a total of 361 ambulatory men aged 65 to 98 with completed data were enrolled for the final analysis.Tianliao township, with 23.7% of its residents aged over 65, is a rural community located in Kaohsiung County, southern Taiwan, containing only one primary care unit. The Tianliao Old People (TOP) study has been conducted in an aged cohort since 2009 in order to develop better community-oriented primary care (COPC) services. According to the 2010 census registered data, there were 1,033 elderly men aged over 65 in Tianliao township. An epidemiological survey using whole community sampling method was performed in July 2010. After excluding empty houses (n = 269), death (n = 21) and non-ambulatory subjects (n = 62), only 681 subjects were eligible and 414 subjects were enrolled in the study. The response rate was 60.8% and the statistical power was 0.80. There were no statistically significant differences in mean age  and each subject signed the inform consent before examination.Each subject was interviewed by well-trained research assistants and completed 20-minute structured questionnaires Habitual tea, alcohol or coffee drinking was defined as those who consumed tea, alcohol or coffee more than once a week for more than half a year. The questionnaire for tea consumption included current drinking habit, tea types, duration and daily amount of tea. The question were “Have you drunk tea habitually once a week for at least 6 months?” Subjects who answered “yes” were coded as habitual tea drinkers and completed the following questions. 1) Have you drunk tea habitually now? 2) How many years have you been drinking tea in this way? 3) What kind of tea  was mostly consumed? 4) How often do you drink tea each week or day? 5) How much (milliliters) tea do you drink each day? Finally, the average amount of daily tea consumption (milliliters) was calculated [(days x volume of tea extracts each day)] The subjects’ smoking habits were obtained from the questionnaire, including current smoking habits, history of quitting smoking, duration of smoking habit and average daily cigarette consumption. A current smoker was defined as a subject who had smoked more than 100 cigarettes and was still smoking Marital status was defined as ‘coupled’ if the subject was married and still lived with their partner 2) was then calculated. Standing naturally, looking forward and wearing only their underwear, the subjects’ waist circumferences (WC) were measured midway between the lateral lower rib margin and the superior anterior iliac crest with a standard tape  at the end of a gentle expiration phase With the subject wearing light clothing without shoes, body weight  and body height  were all measured and the body mass index , triglyceride, uric acid (UA), fasting insulin, and high sensitivity C-reactive protein (hs-CRP) p<0.05 for two-tailed analysis.All continuous variables, including age, BMI, WC, TSA and laboratory data, were expressed as means (SD). The dichotomous data, such as socio-demographic data, habitual behaviors including alcohol drinking, tea consumption habit, different smoking habits and MetS, were expressed as percentages. Data with missing values were excluded when analyzed. The authors used a log-transformation method for the HOMA index and hs-CRP to fit the normal distribution model. Binary logistic regression analysis was used to assess the independent contribution to MetS by possibly associated factors that shows statistical significance in univariate analysis or had been emphasized in previous studies, including age, BMI, UA, log HOMA index, log hs-CRP, occupational status, marital status, educational status, alcohol and coffee drinking habits, tea consumption habit, smoking habit and physical activity. To evaluate the independent effect of smoking and tea drinking habits on MetS, three logistic regression models were formulated. In Model I, the current effects of smoking status  and different types of tea drinking habits on MetS was analyzed. In Model II, the cumulative effects of smoking habit  and tea drinking habit (grouped by less than or over 10 years) was analyzed. In Model III, the daily consumption of cigarette smoking habit  and tea drinking habit  will be analyzed subsequently. All analyses were performed using the Statistical Package of Social Science for Windows software Version 16 . Statistical significance was defined as 2, respectively. One hundred and thirty-two (36.6%) of the subjects met the MetS diagnostic criteria. Compared with the non-MetS subjects, those with MetS had relatively higher BMI, WC, heavy smoker defined by TSA, UA and HOMA indices, but relatively lower levels of physical activity (The average age of the 361 subjects was 74.7±6.1. The mean WC and BMI were 86.4±10.6 cm and 24.1±3.1 kg/mactivity . The intactivity .p>0.05), and the Nagelkerke and Cox & Snell R squares were calculated. All these three models revealed that BMI, UA and HOMA index are significantly associated factors for MetS in the elderly men living in rural community. Model I focused on the current effects of smoking status and different types of tea drinking habits. In model I, after adjustment for age, BMI, UA, HOMA index, hs CRP, physical activity, psycho-social factors , alcohol and coffee drinking habits, the current tea drinker who consumed un- or partial fermented tea habitually had significant negative whilst the current smoker had positive association with MetS compared with those who were non- tea drinker or non-smoker  had a significantly higher prevalence of MetS compared with non-smokers. Those who had a current tea drinking habit  had a significantly lower prevalence of MetS compared with non-current tea drinkers. The results also showed evidence of cumulative dose-effects of smoking and tea drinking habits. To the best of our knowledge, this is the first community-based study that demonstrates the interrelationships among MetS, TSA and habitual tea consumption.To identify previous related works, we used literature sources from Ovid MEDLINE with an English-language search from 1996 to December 2011 using the following search terms: aged, elderly, male, MetS, smoking habit, and tea drinking habit. The association between smoking habits and MetS is inconclusive in the literature, and this is probably due to differences in the various study designs or the races, ages and populations of the subjects Although the pathophysiology of MetS is not conclusive, there is evidence that insulin resistance is the common soil p = 0.08) in association with MetS. The same result had been reported in another cross-sectional study in discussing the relationship between tea and hypertension The negative association between tea drinking and MetS was clearly demonstrated, especially for current tea drinkers who drinking un- or partial fermented tea (green tea or oolong tea), for less than 10 years and more than 240 cc daily. Though data failed to show significant association for drinking tea for more than 10 years, but there was trend to have lower OR (Consistent with previous reports, our study further found a number of associated factors of MetS for elderly male Taiwanese, including hyperuricemia, higher BMI and HOMA indices. Many researchers have concluded the hyperuricemia is associated with the development of MetS Many studies have reported that BMI is significantly associated with MetS and is useful for predicting the onset of MetS and similar results were found in the current study Our study showed physical activity is significant associated with MetS only in univariate analysis. Studies reported inverse association between MetS and physical activity in the elderly r. Finally, our study focused on elderly males living in a rural area with a high prevalence of MetS, and care should thus be taken when extrapolating the findings across a wider age range or more variety of population.Our study has some potential limitations. First, the issue of recall bias may be significant, as it often appears with more complicated behavior or when something happened 20–30 years ago. However, for simple imprinted events, such as history of diseases, educational status and long-term life styles, the recall bias may be minimized Further research may include an outcome study that focuses on the long term effects of habitual tea drinking and smoking habits on CVD and all cause mortality. Because the relationships among the key elements of tea drinking and smoking habits with MetS are still inconclusive, additional research is needed.In conclusion, our study suggests that current smokers, especially those with a higher TSA and daily cigarette amount, are significantly positive associated with MetS. In contrast, current tea drinkers, especially for those subjects who had drunk tea for moderate duration and un- or partial fermented tea with more than 240 cc daily might have a negative association with MetS in elderly males dwelling in a rural community. As the growing epidemic of MetS and tobacco smoke exposure will synergistically increase the risk of CVD


The functional importance of biological entities makes their understanding, analysis, and representation essential in modern biology. Arguably, semantic representation necessary for machine interoperability is a far more difficult task than syntactic representation, necessitating conceptual schema and ontologies for in-silico biological knowledge representation. Biological ontologies are increasingly being developed for prediction, big data integration in semantic web, visualization, unstructured data interpretation, annotation, and eHealth ontology. Despite being widely used, deficiencies exist , whereas a conceptual entity is necessarily dependent on mind, my conjecture is that identity criteria has been violated. Using the DOLCE top-level ontological distinctions, and reorganizing conceptual entity as an agentive-physical-object (DOLCE:APO) and organism attribute as a non-agentive-physical-object (DOLCE:NAPO) could have helped to detect such inconsistencies.In the GO, the term “GO:0020037:heme binding” is a molecular function. From Guarino, , 2002, Ihttp://obo.sourceforge.net), relations lack explicit formal definitions creating the possibility of confusions. Inadequacies could also be found in the use of relations such as is_A and part_Of  (Biological ontologies are plagued by deficiencies in conceptual integration and inter-linkage (Beisswanger et al.,


Recent work has revealed a relationship between pathological video game use and increased impulsivity among children and adolescents. A few studies have also demonstrated increased risk-taking outside of the video game environment following game play, but this work has largely focused on one genre of video games . Motivated by these findings, the aim of the current study was to examine the relationship between pathological and non-pathological video game use, impulsivity, and risky decision making. The current study also investigated the relationship between experience with two of the most popular genres of video games  and risky decision making. Consistent with previous work, ~7% of the current sample of college-aged adults met criteria for pathological video game use. The number of hours spent gaming per week was associated with increased impulsivity on a self-report measure and on the temporal discounting (TD) task. This relationship was sensitive to the genre of video game; specifically, experience with FPS games was positively correlated with impulsivity, while experience with strategy games was negatively correlated with impulsivity. Hours per week and pathological symptoms predicted greater risk-taking in the risk task and the Iowa Gambling task, accompanied by worse overall performance, indicating that even when risky choices did not pay off, individuals who spent more time gaming and endorsed more symptoms of pathological gaming continued to make these choices. Based on these data, we suggest that the presence of pathological symptoms and the genre of video game  may be important factors in determining how the amount of game experience relates to impulsivity and risky-decision making. Past research has demonstrated that video games experience influences cognition and emotion in multiple ways  from Iowa State University ranging in age from 16 to 30 years. Due to an error in the software, data for the testing phase of the probabilistic selection task was lost for one participant. Informed consent was obtained from all participants and they received course credit for their participation. The study was approved by the Institutional Review Board of the university.The media usage questionnaire included three higher order questions. Two questions asked the individual to indicate the number of hours spent playing video games on a typical weekday  or weekend  for each of four time periods . The third question asked the participant to indicate how often s/he plays each of 12 different genres of video games and what video game they spent the most time playing. The dependent variables used were the total number of hours spent playing video games per week and classification as an FPS or strategy video game player (0 or 1) based on the genre of the video game they reported playing most often. The internal reliability was high for the number of hours played (coefficient α = 0.85) and for the amount of experience with genres of video games (coefficient α = 0.87).A revised version of the PVP scale Gentile,  was compThe Barratt Impulsiveness Scale Version 11 . Participants signed the informed consent and completed the BIS-11, pathological gaming scale, RAS, and the media usage questionnaire. Half of the participants completed the tasks in the following order: TD, risk task, Iowa Gambling Task, and Probabilistic Selection; the other half of the participants completed the tasks in the reverse order. Participants also completed the useful-field of view and stop-signal tasks, but as these data do not specifically address the relationship between video games and risky decision making, it is not reported here. After the tasks were completed the participants were debriefed and thanked for their participation. The entire study took ~90 min.SD = 25.4, 25th quartile = 0, 50th quartile = 13, 75th quartile = 34). Males reported playing more hours per week  than females , t(147) = 4.06, p < 0.001. Pathological gaming  was reported by 7.4%  of the sample, consistent with the rate observed in other samples of children and adolescents  than females , t(147) = 6.90, p < 0.001.Table Gentile, . The meaCorrelations among all of the variables included in the analyses are presented in Table M = 0.79, SD = 0.17; non-FPS gamers: M = 0.71, SD = 0.22; t(147) = −2.10, p = 0.04] and Hours × FPS . Self-reported impulsivity was positively correlated with Hours and Hours × PVP, consistent with previous work  avoided B more frequently than non-strategy gamers , t(146) = −2.09, p = 0.04, supporting the hypothesis that this genre may encourage players to learn from mistakes and avoid making them in the future.Selection from bad decks in the IGT was positively correlated with Hours × PVP Figure , supportR2c) of 0.39, 0.28, 0.19, 0.14, 0.11, 0.05, 0.03, 0.02, and 0.01 for functions one through nine, respectively. The full model was significant using the Wilks's λ = 0.25 criterion, F = 1.68, p < 0.001. Wilks's λ represents the variance unexplained by the model, therefore 1—Wilks's λ represents the full model effect size in terms of r2. In this analysis with nine canonical functions, the r2 was 0.75, indicating that the full model explained 75% of the variance between the two sets of variables. To test the hierarchical arrangement of the functions for statistical significance, a dimension reduction analysis was used , and risky decision making  and the functions indicate which variables have the strongest contribution to the function and can be interpreted in a similar manner as the factor loadings in a factor analysis  and greater impulsivity Figure .The current study was designed to examine the relationships between video game experience, pathological gaming, and risky decision making. Consistent with previous work Gentile, , ~7% of The self-report and behavioral measures revealed that pathological gaming and playing FPS games were positively associated with greater impulsivity. The interaction between hours and PVP was also positively related to BIS-II scores indicating that more pathological symptoms were positively associated with greater impulsivity  appeared to be higher than the correlation between PVP and hours × FPS (r = 0.29), although this difference did not reach significance, t(146) = 1.53, p > 0.05. This suggests that both strategy and FPS games are associated with pathological gaming, but that the consequences for impulsivity and risk-taking are not the same for the two genres. This may be due to the structure of the gaming environment or the goals of the players within the different genres.The association between gaming and risky decisions was sensitive to game genre. In the risk task, the number of hours spent playing video games, the interaction between hours and PVP, and categorization as an FPS gamer were all negatively correlated with the percentage of low risk selections; and hours predicted selecting the high-risk option more frequently in the CCA. This was accompanied by a dramatic reduction in the total points earned at the end of the task, indicating that the selection of risky options in the risk task did not pay off in the end. Taken together, these findings provide clear evidence that gaming time, pathology, and FPS games influence an individual's selection of risky options, and this behavior appears to continue in spite of its detrimental effect on performance over time. In contrast to FPS gaming, strategy gaming was not as strongly related to increased risk-taking. One explanation for the differential influence of game genre is that there are likely to be social repercussions for making impulsive decisions in a strategy game since successful in the game often requires cooperation with a team. It is important to note that both strategy games and FPS games were positively correlated with PVP and the correlation between pathological symptoms and hours × strategy  due to greater selection of risky options. Presumably, after several selections of the low chance risky options, the accruement of losses should be a deterrent for further selection of the risky option, but this did not appear to be the case. Similarly, feedback over several trials of the IGT should result in decreased selection from the “bad” decks. Higher PVP scores and hours gaming were associated with greater selection from the “bad” decks well past the point at which the feedback was effective at decreasing selection from these decks among non-pathological high gamers.r = 0.17), suggesting that individuals who identify as strategy gamers are more sensitive to negative feedback. As with impulsivity, the characteristics of strategy games may explain this relationship. Mistakes in a strategy game can have long-term consequences for achieving goals in the game because game play usually spans a longer time frame than an FPS video game. Therefore, mistakes in a strategy video game can be costly, and one would benefit from paying attention to negative outcomes and learning to avoid those outcomes in the future.The probabilistic selection task  account for the findings, and the direction of causation cannot be defined . Future studies may address this issue by looking at the short- and long-term effects of video game training on risky decision making, similar to work on aggression (Anderson et al., The current study extends the literature on the relationship between video game experience and risky decision making beyond risky driving behavior (Fischer et al., The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


WNT10A-related ectodermal dysplasias. The ectodermal dysplasias (ED) are a large, heterogeneous group of disorders characterized by defects in the morphogenesis of skin, sweat, sebaceous, submucous, and mammary glands, hair, nails and teeth. Numerous more or less distinct entities have been reported, however, most of them are very rare and the cause is often unknown. EDA mutations, resulting in hypohidrotic ectodermal dysplasia (Christ Siemens Touraine syndrome) are known as the most common cause of ED. Over the last two years, WNT10A has turned out to be the second major gene associated with ED. WNT10A mutation carriers present with a broad and variable spectrum of symptoms related to at least four known Mendelian conditions as there are autosomal dominant Selective Tooth Agenesis Type 4 and autosomal recessive Adontia of Permanent Dentition (APD), Odonto-Onycho-Dermal Dysplasia (OODD), and Schöpf-Schulz-Passarge syndrome (SSPS). In heterozygotes, penetrance is reduced and only about half of the carriers show a phenotype including mainly tooth and nail anomalies. Approximately 8% of the unaffected individuals in our control group carried a WNT10A mutation. Further studies are necessary. We conclude that WNT10A mutation analysis might become an important diagnostic test in many patients with selective tooth agenesis or severe oligodontia concerning the permanent teeth.Here we report on the clinical findings and key symptoms in


Models of competitive template replication, although basic for replicator dynamics and primordial evolution, have not yet taken different sequences explicitly into account, neither have they analyzed the effect of resource partitioning (feeding on different resources) on coexistence. Here we show by analytical and numerical calculations that Gause's principle of competitive exclusion holds for template replicators if resources (nucleotides) affect growth linearly and coexistence is at fixed point attractors. Cases of complementary or homologous pairing between building blocks with parallel or antiparallel strands show no deviation from the rule that the nucleotide compositions of stably coexisting species must be different and there cannot be more coexisting replicator species than nucleotide types. Besides this overlooked mechanism of template coexistence we show also that interesting sequence effects prevail as parts of sequences that are copied earlier affect coexistence more strongly due to the higher concentration of the corresponding replication intermediates. Template and copy always count as one species due their constraint of strict stoichiometric coupling. Stability of fixed-point coexistence tends to decrease with the length of sequences, although this effect is unlikely to be detrimental for sequences below 100 nucleotides. In sum, resource partitioning (niche differentiation) is the default form of competitive coexistence for replicating templates feeding on a cocktail of different nucleotides, as it may have been the case in the RNA world. Our analysis of different pairing and strand orientation schemes is relevant for artificial and potentially astrobiological genetics. The dynamical theory of competing templates has not yet taken the effect of sequences explicitly into account. One might think that complementary sequences have very limited competition only. We show that, despite interesting sequence effects, competing template replicators yield to Gause's principle of competitive exclusion so that the number of stably coexisting template species cannot exceed the number of nucleotide species on which they grow, although one of the findings is that plus and minus strands together count as one species. Thus up to four different templates/ribozymes can constitute the first steps to an early, segmented genome: we suggest that other mechanisms build on this baseline mechanism. Gause (1934) in the Golden Age of theoretical ecology formulated the principle of competitive exclusion, proposing in effect what usually is being referred to as “complete competitors cannot coexist” We explicitly take into consideration the concentration of up to four different building blocks  and that the dynamics of these abiotic resources are not periodically forced, for example. We neglect replicase enzymes and assume that template and replica separate irreversibly upon completion of elongation. The kinetic effects are simplified to the extent that the elongation rate of template polymerization depends only on the identity of the inserted nucleotide and nothing else. We know that this is a crucial simplification but already with this rule different sequences may assume very different kinetic phenotypes. In agreement with this, we neglect secondary and tertiary structures.raison d'être for these assumptions is that we would like to demonstrate the effect of competition for resources of competing template sequences as simply and clearly as possible.  We deliberately want to see the dynamics of coexistence under irreversible exponential growth tendency, as a kind of worst case. The effect of the sequence diversity of templates on dynamical coexistence is not trivial. If there are two resources A and B, then it is trivial that sequences of The Some of the effects that we show in this paper are far from trivial. Our calculations show the effect of resource partitioning on template coexistence and shed more light on early molecular evolution, which surely was affected by sequence effects of template replicators.sequences) we formulated the dynamics of polynucleotide replication. Here we only explain the necessary basics of our formalism, for the mathematical model see To understand the mechanism of coexistence of template replicators ; we also introduce parallel strand polarity as opposed to antiparallel polarity (like in case of RNA replication). The difference between complementary and non-complementary pairing and parallel and antiparallel strand polarity is given in The following model of template replication models RNA replication, dealing with 4 monomers, complementary pairing and antiparallel strand polarity. We have investigated the coexistence of M1 With a given set of parameters (elongation constants M2 We have investigated the coexistence for each sampled sequence group for different numbers of complementary pairs (and stability of coexistence and that on four different monomers a maximum of four different sequence pairs could possibly coexist. Thus Gause's principle (against first intuition) limits the number of sequence pairs instead of individual sequences because of the dynamical coupling between the template and its complement . According to our exhaustive numerical results, we have found no case where more than two sequences could coexist, supporting our hypothesis. We have numerically integrated ODE systems until convergence or extinction of one of the sequences. The numerical methods and routines are the same as before. We have analyzed the coexistence of two sequences in two different ways  of coexistence of sequences can be deduced right from the order of monomers of the sequences according to a very simple rule . The rulA-s in its head; the latter behaves as having A-s in the head to yield coexistence  weighs more concerning the coexistence than the rear (tail). This is because during replication, earlier intermediates are present in larger concentrations than intermediates closer to the final step of the replication  the trinculeotide Mechanisms for template coexistence have been in the focus of models of primordial replicator evolution cf. . Here weRecently there has been an upsurge in interest in exo/astrobiology. It is in this context that we have deliberately presented results for homologous pairing also, even with parallel orientation of the strands. Although such configurations are not unheard of even in our world, we wanted to see how such features would in general affect dynamical coexistence of template replicators.We have obtained the fitness landscapes through a distribution of elongation and degradation rates. The main reason behind this is tractability: although the 2D structures as phenotypes of RNA molecules can be calculated for most cases, this does not automatically yield phenotypes in terms of replication rates. We are temporarily satisfied with the phenotype richness that our local rules provided see . What isIn each experiment, we integrated the system of Let us introduce two complementary sequences:The extension of the dynamics for more sequence pairs  is straightforward. The dynamics of the intermediates is independent for each pair and the dynamics of the monomers provides the coupling between the equations of different pairs of sequences. Because of the cross-coupling of equations, no analytical solution was found . For the numerical integration of the ODE system to find steady-state solutions we have used the CVODE code from the SUNDIALS project of the Lawrence Livermore National Laboratory Uniform degradation rates of sequence intermediates allow for a completely analytic approach. For the positivity test of concentrations, we introduce the following notation for the constants of the power sum of different signs.Let us assume that influx can counter degradation. In this case the condition of coexistence  of length . Lower left half: coexistence is marked by green, extinction of the first sequence pair by red and extinction of the second sequence pair by blue. Upper right half: stability of coexistence according to the leading eigenvalue . The upper triangle shows the stability measures of the sequences pairs from the lower one (mirrored and rotated (TIF)Click here for additional data file.Figure S2Coexistence plots of pairs of double-stranded sequences of length  (upper panel) and  (lower panel) using two monomers  in case of uniform degradation and identical elongation rate constants and non-complementary pairing. The green indicates stable coexistence, grey indicates structurally unstable coexistence, i.e. compositional identity , while pink indicates that there is no coexistence possible. Sequences along the axes are arranged first according to Hamming distance and secondly according to lexicographic ordering from more (TIF)Click here for additional data file.Figure S3Correlation plot of the fitness landscape as a function of Hamming distances for sequence pairs of length .(TIF)Click here for additional data file.Table S1Analysis of coexistence of  non-complementary pairs of sequences of length  according to method M1. Second column shows the number of scanned sequences (and the amount as a fraction of the whole combined sequence space). The third column shows the fraction of coexisting sequences in the scanned domain, i.e. the probability of coexistence of random sequence groups of size (XLS)Click here for additional data file.Table S2Analysis of coexistence of  non-complementary pairs of length  according to method M2. Second column shows the number of scanned sequences (and the amount as a fraction of the combined sequence space). The third column shows the fraction of coexisting sequences averaged over the scanned sequence groups and over 1000 random degradation rate sets for intermediates Click here for additional data file.Table S3Examples of coexistence for .(XLS)Click here for additional data file.Table S4Examples of coexistence for .(XLS)Click here for additional data file.Table S5Examples of coexistence for .(XLS)Click here for additional data file.Text S1Supporting text with sections on 1) Parameters for methods M1, M2, M3 and M4; 2) Analysis of non-complementary pairing with antiparallel polarity; 3) Analysis and analytical results of non-complementary pairing and uniform degradation rates; 4) Proofs; 5) Discussion of the fitness landscape; 6) Examples of coexistence of longer sequences.(PDF)Click here for additional data file.