Peroxisome proliferator-activated receptor gamma (PPARγ), a nuclear receptor, has been implicated playing a role in the development of inflammatory bowel disease (IBD). However, previous studies evaluating the association between the PPARγ2 Pro12Ala polymorphism and IBD are inconsistent. We performed a meta-analysis to determine whether the PPARγ2 Pro12Ala mutation was associated with the presence of IBD.P = 0.162, Bon = 1.000; ProPro vs ProAla: OR = 1.042, 95%CI = 0.852–1.273, P = 0.690, Bon = 1.000; ProPro vs AlaAla: OR = 2.379, 95%CI = 1.110–5.100, P = 0.026, Bon = 0.156; ProAla vs AlaAla: OR = 2.315, 95%CI = 1.064–5.037, P = 0.034, Bon = 0.204; Pro homozygotes vs Ala positives: OR = 1.094, 95%CI = 0.899–1.330, P = 0.371, Bon = 1.000; Pro positives vs Ala homozygotes: OR = 2.360, 95%CI = 1.103–5.053, P = 0.027, Bon = 0.162; heterozygotes vs all homozygotes: OR = 0.976, 95%CI = 0.799–1.192, P = 0.809, Bon = 1.000). There was no significant association of this polymorphism with UC or CD in the East Asian population and the Turkish population.Electronic databases were searched for case-control studies evaluating the association between the Pro12Ala mutation and the presence of IBD. Effects were summarized with the methods recommended by the Cochrane Collaboration. A total of 7 studies including 1002 ulcerative colitis (UC) cases, 1090 Crohǹs disease (CD) cases and 1983 controls were involved in this meta-analysis. In the overall analysis, no significant association of this polymorphism with UC or CD was found. In the subgroup analyses in different populations, AlaAla genotype seemed to protect the European Caucasian population against the development of CD (Pro vs Ala: OR = 1.135, 95%CI = 0.951–1.354, AlaAla genotype may be a protective factor in the European Caucasian population against the development of CD in a recessive way. Inflammatory bowel disease (IBD) clinically classified into ulcerative colitis (UC) and Crohǹs disease (CD) is a non specific chronic intestinal inflammatory disorder with the characteristics of relapses and remissions. When diagnosed, patients with UC or CD need lifelong medications. Moreover, strictures, abscesses, fistulas, extra intestinal involvements and even carcinomas would complicate IBD and eventually affect the progression of IBD. In the last three decades, we have witnessed a time trend increase of the prevalence of IBD especially of CD both in developed and developing regions Although the treatment of IBD has advanced considerably and guidelines have just been revised, IBD is still incurable Peroxisome proliferator-activated receptor gamma (PPARγ), a nuclear receptor, has been implicated as playing some role in the development of UC and CD PubMed (1966 to June 2011) and Scopus (1966 to June 2011) were searched using the combinations of text words and medical subject headings (MeSH) including “inflammatory bowel disease”, “ulcerative colitis”, “Crohn's disease”, “IBD”, “UC”, “CD”, “peroxisome proliferator-activated receptor”, “PPAR”, “polymorphism”, “polymorphisms”, “single nucleotide”, “allele” and “genotype”. Chinese medical databases including Wanfang database (1982 to June 2011), China National Knowledge Infrastructure  and Chinese Biomedical Literature Database  were also searched for relevant articles. We searched reference lists, relevant meta-analyses and reviews in order to get additional articles.The following selection criteria were employed in this meta-analysis: 1) Case-control studies evaluating the association between the PPARγ2 Pro12Ala mutation and the presence of IBD. 2) Diagnosis of IBD according to clinical manifestations, radiological changes, endoscopic manifestations, and histological evaluations comprehensively. 3) Hardy-Weinberg equilibrium fulfilled in the control arm of each research. 4) Articles with a full text not just an abstract. 5) Articles providing raw data or Odds ratio (OR) and its 95% confidence interval (CI) for each comparison. Exclusion criteria were as the follows: 1) Repetitive publications. 2) Family based case-control studies.Two reviewers screened the titles and abstracts of potentially relevant articles. And the full texts of highly relevant articles would be thoroughly read by two reviewers. When a discrepancy was encountered, a third reviewer would be referred to and the decision would be made through discussions. Publication year, region the study was conducted in, ethnicity, available allele and genotype frequencies in each arm and the method of polymorphism assessment of each study were exacted.We also assessed the potential risks of bias with the items including 1) Selection bias , 2) Information bias  and 3) confounding factors (cases and controls matching).2I statistics was also used to evaluate the risks of heterogeneity among studies: 0%–40% represented no risk of heterogeneity, 30%–60% represented a low risk of heterogeneity, 50%–90% represented substantial heterogeneity and 75%–100% represented considerable heterogeneity P value was more than 0.1, the 2I statistics indicating no or a low risk of heterogeneity and the CI of each study overlapped, a fixed model was employed and the Mantel-Haenszel method was implemented to synthesize data. If the P value was less than 0.1, the 2I statistics indicating substantial or considerable heterogeneity or the CI of each study did not overlapped, a random model was employed and the DerSimonian-Laird method was applied to synthesize data. Funnel plots were used to examine bias in the results of the meta-analyses http://cran.csdb.cn/) was used for the step down Bonferroni adjustments (Bon). Values of P<0.05 were considered statistically significant. All the above methods had been used for pooling data in two previous published genetic association meta-analyses [16.17].Hardy-Weinberg equilibrium was assessed using the χ2 test. Paired combinations of genotypes were employed to determine the hereditary models: 1) an allelic analysis (Pro vs Ala); 2) a genotypic analysis  and 3) another genotypic analysis comparing each genotype with the other two . If the comparisons of ProPro vs AlaAla, ProPro vs ProAla and Pro homozygotes vs Ala positives were statistically significant, the Ala allele would be a dominant allele. If the comparisons of ProPro vs AlaAla, ProAla vs AlaAla and Pro positives vs Ala homozygotes were statistically significant, the Ala allele would be a recessive allele. OR and its 95% CI were calculated with the methods recommended by the Cochrane Collaboration, and a fixed value of 0.5 was added to all cells of study results tables where no events were observed in one or both groups 91 articles were identified during premature searches with our searching strategy of the five databases . 59 articles were retrieved after excluding overlapping studies. After excluding reviews, animal studies, comments, letters and studies not evaluating IBD, 16 studies assessing the association between the PPARγ polymorphism and IBD were found. Among the remaining 16 articles, 8 studies did not evaluate the Pro12Ala mutation and one republication was identified. Only 7 articles met our selection criteria. After searching reference lists, relevant meta-analyses and reviews, we did not find additional studies. The selection process is illustrated in Selection bias: Cases were selected from diagnosed UC and CD patients in medical institutions in all the included studies. 3 studies recruited healthy blood donors as controls Information bias: 3 studies used polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) as the single nucleotide polymorphism (SNP) detection method Confounding factors: Age and gender distribution were comparable among arms in most studies, while controls were slightly older as compared to CD patients in one study 2I statistics and the detailed meta-analysis results are shown in The CI overlapping status, Cochrane Q χ2 tests, P = 0.486, Bon = 1.000; ProPro vs ProAla: OR = 1.088, 95%CI = 0.883–1.340, P = 0.429, Bon = 1.000; ProPro vs AlaAla: OR = 0.644, 95%CI = 0.347–1.197, P = 0.165, Bon = 1.000; ProAla vs AlaAla: OR = 0.606, 95%CI = 0.320–1.149, P = 0.125, Bon = 0.875; Pro homozygotes vs Ala positives: OR = 0.968, 95%CI = 0.727–1.289, P = 0.823, Bon = 1.000; Pro positives vs Ala homozygotes: OR = 0.625, 95%CI = 0.337–1.159, P = 0.136, Bon = 0.952; heterozygotes vs all homozygotes: OR = 0.910, 95%CI = 0.739–1.120, P = 0.372, Bon = 1.000). Between-study heterogeneity was detected in the contrasts of Pro vs Ala and Pro homozygotes vs Ala positives. The association between the PPARγ Pro12Ala polymorphism and CD was investigated in 6 studies with 1090 CD cases and 1841 controls P = 0.204, Bon = 1.000; ProPro vs ProAla: OR = 1.024, 95%CI = 0.845–1.242, P = 0.807, Bon = 1.000; ProPro vs AlaAla: OR = 1.918, 95%CI = 0.938–3.922, P = 0.074, Bon = 0.518; ProAla vs AlaAla: OR = 1.986, 95%CI = 0.953–4.139, P = 0.067, Bon = 0.469; Pro homozygotes vs Ala positives: OR = 1.075, 95%CI = 0.890–1.297, P = 0.454, Bon = 1.000; Pro positives vs Ala homozygotes: OR = 1.906, 95%CI = 0.933–3.893, P = 0.077, Bon = 0.539; heterozygotes vs all homozygotes: OR = 0.991, 95%CI = 0.818–1.201, P = 0.927, Bon = 1.000). Between-study heterogeneity was not detected in all the contrasts.The association between the PPARγ Pro12Ala polymorphism and UC was investigated in 6 studies with 1002 UC cases and 1867 controls P = 0.651, Bon = 1.000; ProPro vs ProAla: OR = 1.157, 95%CI = 0.917–1.459, P = 0.219, Bon = 1.000; ProPro vs AlaAla: OR = 0.680, 95%CI = 0.346–1.335, P = 0.263, Bon = 1.000; ProAla vs AlaAla: OR = 0.586, 95%CI = 0.293–1.173, P = 0.131, Bon = 0.917; Pro homozygotes vs Ala positives: OR = 1.109, 95%CI = 0.886–1.388, P = 0.365, Bon = 1.000; Pro positives vs Ala homozygotes: OR = 0.653, 95%CI = 0.334–1.280, P = 0.215, Bon = 1.000; heterozygotes vs all homozygotes: OR = 0.854, 95%CI = 0.678–1.077, P = 0.183, Bon = 1.000). Between-study heterogeneity was not detected in all the contrasts. The association between the PPARγ Pro12Ala polymorphism and CD in the European Caucasian population was investigated in 3 studies with 979 CD cases and 1387 controls P = 0.162, Bon = 1.000; ProPro vs ProAla: OR = 1.042, 95%CI = 0.852–1.273, P = 0.690, Bon = 1.000; ProPro vs AlaAla: OR = 2.379, 95%CI = 1.110–5.100, P = 0.026, Bon = 0.156; ProAla vs AlaAla: OR = 2.315, 95%CI = 1.064–5.037, P = 0.034, Bon = 0.204; Pro homozygotes vs Ala positives: OR = 1.094, 95%CI = 0.899–1.330, P = 0.371, Bon = 1.000; Pro positives vs Ala homozygotes: OR = 2.360, 95%CI = 1.103–5.053, P = 0.027, Bon = 0.162; heterozygotes vs all homozygotes: OR = 0.976, 95%CI = 0.799–1.192, P = 0.809, Bon = 1.000). Between-study heterogeneity was not detected in all the contrasts.The association between the PPARγ Pro12Ala polymorphism and UC in the European Caucasian population was investigated in 2 studies with 598 UC cases and 1271 controls P = 0.353, Bon = 1.000; ProPro vs ProAla: OR = 0.856, 95%CI = 0.502–1.460, P = 0.568, Bon = 1.000; ProPro vs AlaAla: OR = 0.490, 95%CI = 0.089–2.700, P = 0.413, Bon = 1.000; ProAla vs AlaAla: OR = 0.774, 95%CI = 0.128–4.669, P = 0.780, Bon = 1.000; Pro homozygotes vs Ala positives: OR = 0.686, 95%CI = 0.284–1.654, P = 0.401, Bon = 1.000; Pro positives vs Ala homozygotes: OR = 0.500, 95%CI = 0.091–2.750, P = 0.425, Bon = 1.000; heterozygotes vs all homozygotes: OR = 1.164, 95%CI = 0.683–1.984, P = 0.577, Bon = 1.000). Between-study heterogeneity was detected in the contrasts of Pro vs Ala and Pro homozygotes vs Ala positives. The association between the PPARγ Pro12Ala polymorphism and CD in the East Asian population was investigated in 2 studies with 42 CD cases and 354 controls P = 0.886, Bon = 1.000; ProPro vs ProAla: OR = 0.933, 95%CI = 0.289–3.012, P = 0.907, Bon = 1.000; ProPro vs AlaAla: OR = 0.324, 95%CI = 0.029–3.663, P = 0.363, Bon = 1.000; ProAla vs AlaAla: OR = 0.458, 95%CI = 0.031–6.825, P = 0.571, Bon = 1.000; Pro homozygotes vs Ala positives: OR = 1.011, 95%CI = 0.315–3.249, P = 0.985, Bon = 1.000; Pro positives vs Ala homozygotes: OR = 0.321, 95%CI = 0.028–3.623, P = 0.358, Bon = 1.000; heterozygotes vs all homozygotes: OR = 1.082, 95%CI = 0.335–3.493, P = 0.896, Bon = 1.000). Between-study heterogeneity was not detected in all the contrasts.The association between the PPARγ Pro12Ala polymorphism and UC in the East Asian population was investigated in 3 studies with 359 UC cases and 496 controls The association between the PPARγ Pro12Ala polymorphism and IBD in the Turkish population was investigated in 1 study with 45 UC cases, 69 CD cases and 100 controls Moreover, the conclusions of our meta-analysis did not change when we used a fixed model and a random model to perform our meta-analysis respectively.Obvious asymmetry was revealed by the shapes of the Begg funnel plots in the overall UC analysis and the overall CD analysis. The study with the smallest sample size reported by Aoyagi was a clear outlier which might be suggestive of a small-study effect. The Begg funnel plots corresponding to the allelic analysis are shown in The core pathophysiological process of UC is the uncontrolled inflammation mainly affecting the colon. Environmental factors including commensal flora and other antigens and host factors including genetic dispositions are all involved in the UC development Firstly, dysfunction of the cellular and immune intestinal barrier which separates the microbial flora from host tissues renders a host susceptible to invasive factors PTPN2, IL18RAP, TAGAP, and PUS10 loci as new CD risk factors The mechanism of the uncontrolled inflammation in CD is different from that in UC. The deficiency of the innate immune system which permits intestinal pathogens to break through the intestinal barrier and activates the adaptive immune system may be responsible for the uncontrolled inflammation in CD Our meta-analysis supports this hypothesis. The AlaAla genotype seemed to protect the European Caucasian population against the development of CD. The Pro12Ala mutation only affects the function of PPARγ2, which supports PPARγ2 as a modulator in the innate immune system maintenance. However, we did not find an association between this polymorphism and CD in the East Asian population. This is in consistence with the observation of NOD2/CARD15 polymorphism on CD in the non Caucasian populations However, the conclusions of this meta-analysis should be interpreted cautiously. This meta-analysis has some limitations. Firstly, only one study used colonoscopy to exclude IBD in the controls In conclusion, based on published full text case-control studies, our meta-analysis demonstrates that the AlaAla genotype may be a protecting factor in the European Caucasian population against the development of CD in a recessive way.Text S1Checklist(DOC)Click here for additional data file.Text S2Scopus searching result(DOC)Click here for additional data file.


Hazardous drinking and depression represent two major sources of harm and dysfunction for university students. This study sought to determine how best to address these frequently co-occurring conditions for students who are not seeking treatment. Participants were recruited through internet ads and screened by phone and in-person to participate in a study on alcohol use, stress, and student life. Thirty university students (aged 18-22) with hazardous drinking who exhibited depressive symptoms were randomly assigned to one of two conditions: assessment only or goal systems intervention (GSI). The GSI is a 3-session motivational intervention that addresses risks and processes of change related to alcohol and depression within an integrated framework. Students completed assessments of depressive symptoms, alcohol consequences, and heavy drinking episodes at baseline, three months, and six months. Given the small sample size of this pilot trial, our main goal was to provide effect-size estimates. Results of the conditional latent growth analysis for alcohol-related negative consequences showed a significant medium-to-large effect of the intervention (f-squared = 0.27). Similarly, analysis showed that the intervention resulted in a medium to large effect on reduction of depressive symptoms (f-squared = 0.323). There was little evidence of intervention effects on change in heavy drinking, as students in both conditions exhibited a significant decline in heavy drinking episodes over time. These results suggest that GSI may be an efficacious intervention to reduce alcohol-related harm and depressive symptoms among university students in opportunistic settings.


Although many genotoxic treatments upregulate the cyclin kinase inhibitor p21, agents such as UV irradiation trigger p21 degradation. This suggests that p21 blocks a process relevant for the cellular response to UV. Here, we show that forced p21 stabilization after UV strongly impairs damaged-DNA replication, which is associated with permanent deficiencies in the recruitment of DNA polymerases from the Y family involved in translesion DNA synthesis), with the accumulation of DNA damage markers and increased genomic instability. Remarkably, such noxious effects disappear when disrupting the proliferating cell nuclear antigen (PCNA) interacting motif of stable p21, thus suggesting that the release of PCNA from p21 interaction is sufficient to allow the recruitment to PCNA of partners (such as Y polymerases) relevant for the UV response. Expression of degradable p21 only transiently delays early replication events and Y polymerase recruitment after UV irradiation. These temporary defects disappear in a manner that correlates with p21 degradation with no detectable consequences on later replication events or genomic stability. Together, our findings suggest that the biological role of UV-triggered p21 degradation is to prevent replication defects by facilitating the tolerance of UV-induced DNA lesions. The levels of the cyclin kinases inhibitor, p21, increases after treatment with various genotoxic agents, including γ irradiation, daunorubicin and others. Increased p21 expression inhibits the activation of cyclin-dependent kinases (CDKs), blocks cell proliferation and promotes cell survival and genomic stability by providing a temporal window for DNA repair  before DAPI stainning.2 UVC and incubated with IdU (200 µM) for additional 20 min. Cells were trypsinized and lysed with 6 µl of 0.5% SDS, 200 mM Tris–HCL (pH 7.4) and 50 mM EDTA buffer onto clean glass slides, which were tilted, allowing DNA to unwind. Samples were fixed in 3:1 methanol/acetic acid and denatured with HCL (2.5 N) for 1 h, blocked in PBS 5% Bovine serum albumin (BSA) for 15 min and incubated with the mouse anti-BrdU (Becton Dickinson) to detect IdU, donkey anti-mouse Cy3-conjugated secondary antibody (Jackson Immuno Research), rat anti-BrdU  to detect CldU and donkey anti-rat Alexa 488 secondary antibody (Invitrogen). Slides were mounted with Mowiol 488 , and DNA fibers were visualized using a Zeiss Axioplan confocal microscope. Images were analyzed using Zeiss LSM Image Browser software. Each data set is derived from measurement of at least 75 fibers.DNA fibers were analyzed using a protocol previously used by us . BrieflyTo evaluate whether the point mutations used disrupted p21/CDK2 and p21/PCNA binding, we performed immunoprecipitations in the fraction (soluble) where p21/CDKs are usually found . Lysatest-test and one-way ANOVA test when applicable. Other calculations and graphics were performed using Microsoft Excel 2007.Frequency distribution of DNA fiber ratios was analyzed with GraphPad Prism 5 software. In non-Gaussian distributions, Mann–Whitney and Kruskal–Wallis tests where used for statistical analysis when comparing 2 and 3 variables, respectively. Statistical analysis of Y pols focal organization was performed in GraphPad inStat software using the Student’s ΔC mutant showed increased stability after UV irradiation (ΔC mutant with a disrupted PCNA interacting protein (PIP) box by introducing point mutations as reported previously . We also assessed the effect of sp21ΔC on the accumulation of 53BP1 foci, which are generally associated with double strand breaks (DSBs) . These results suggest that p21 degradation or the disruption of p21/PCNA interaction efficiently prevents accumulation of replication stress markers after UV irradiation.To explore the effect of forced p21 stabilization after UV irradiation, we evaluated the levels of a s, γH2AX . sp21ΔC, samples F and G, Supplementary Figure S1C–E and our previous report . Interestingly, MN formation was much more frequent in sp21ΔC-expressing cells that were replicating DNA at the time of UV irradiation (EdU+ cells) than in EdU− cells , thus suggesting a global negative effect of p21 on the replication of damaged DNA. Intriguingly, we observed a differential effect of sp21ΔC on Rev1 focal organization. Although Rev1 focal reorganization after UV irradiation was impaired by sp21ΔC . Together, these results indicate that sp21ΔC abolishes the UV-triggered recruitment of all Y family pols to replication factories.We have previously shown that p21 impairs PCNA ubiquitination and the formation of DNA replication-associated foci of pol η ,13, and y sp21ΔC E, a cons nucleus . In contΔC slightly delayed—yet not repressed—pol η focal organization and did not impair pol η/PCNA interaction . When using HCT116p21+/+ and p21−/− cells, we found that before and after stress, the focal organization of most Y pols  was upregulated in HCT116 p21 −/− cells . A shorter second track indicates a delay or a block in the progression of DNA replication after UV irradiation. Average CldU/IdU ratios of ∼1 were obtained in sham-irradiated samples, and UV irradiation shifted the ratios to greater numbers in all samples , it impaired fork progression after UV irradiation . Although this result suggested that, at early time points after UV irradiation, both degradable and stable p21 delays immediate replication events, it is important to highlight that these defects do not propagate equally to later replication events. When analysing cells expressing degradable p21 (either endogenous or exogenously overexpressed), no defects in S phase progression were observed 24 h after UV irradiation . However, sp21ΔC caused persistent defects in S phase progression in a manner that was dependent on PCNA binding . Moreover, when trapping UV-irradiated cycling cells into a binucleated stage , we observed significantly fewer sp21ΔC-expressing cells getting to the binucleated stage . This suggests that sp21ΔC lead to permanent defects (at or before karyokinesis), which might result from irreversible deficiencies in DNA replication that occur when p21 is not removed from PCNA after UV irradiation. Thus, UV-triggered p21 degradation is required to promote damaged-DNA replication after UV irradiation and to preserve genomic stability.Although sp21adiation B. To facadiation C. In thi samples C. In agrwas used D. We conIn this work, we provide evidence indicating that endogenous p21 delays early replication events after UV irradiation, and that its UV-triggered degradation is required to prevent latter defects in S phase progression. Conversely, forced p21 stabilization prevents the interaction of PCNA with Y polymerases complexes, which leads to persistent defects in S phase progression, accumulation of replication stress markers and genomic instability.ΔC expressing cells that were transiting S phase just after UV irradiation is the one that preferentially accumulates micronuclei strongly supports this hypothesis.Many years ago, Bunz and Volegstein showed that p53-dependent p21 upregulation prevents cell death after γ-irradiation by triggering cell cycle arrest . This scAnother intriguing implication of our findings is that endogenous p21 represses—or at least delays—initial TLS events after UV irradiation. We believe that this transient delay in TLS onset might be beneficial for cells. Using a plasmid-based assay, Livneh and colleagues  showed tΔC or sp21ΔC. Although more work is required to elucidate the complex relationship between p21 and PCNA ubiquitination, this article shows that p21 can directly control PCNA/Y polymerase interactions in a manner that is independent of PCNA ubiquitination.p21 might regulate TLS through the control of PCNA ubiquitination ,13. As PΔC specifically prevents the interaction of PCNA with pol η, pol ι and pol κ (this work), without affecting DNA polymerases involved in unperturbed DNA replication  [R03 TW008924]; Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT) (to V.G.); V.G. and G.S. are researchers from CONICET, and S.F.M., M.B.V. and M.H. received fellowships from CONICET and ANPCyT. Funding for open access charge: NIH [R03 TW008924].Conflict of interest statement. None declared.


Cryptosporidium hominis subtypes occurred in a pediatric hospital ward (Ward A) in China, lasting for more than 14 months. In this study, the concurrence during the outbreak of three other enteric pathogens with a similar transmission route, Giardia duodenalis, Enterocytozoon bieneusi, and Clostridium difficile, was assessed.Over 200 cryptosporidiosis outbreaks have been reported, but little is known if other enteric pathogens were also involved in some of these outbreaks. Recently, an outbreak of cryptosporidiosis linked to poor hygiene by two G. duodenalis, E. bieneusi, and C. difficile in 78 inpatients from Ward A and 283 and 216 inpatients from two control wards (Wards C and D) in the same hospital was examined using molecular diagnostic tools. Significantly higher infection rates were found in children in Ward A for all study pathogens than in Wards C and D (P<0.01): 9.5% versus 1.4% and 0% for G. duodenalis, 10.8% versus 2.8% and 3.7% for E. bieneusi, and 60.8% versus 37.8% and 27.8% for C. difficile, respectively. These differences were mostly seen in children ≤12 months. Enteric pathogen-positive children in Ward A (31/58 or 53.4%) were more likely to have mixed infections than those in Ward C (4/119 or 3.4%) or D . Having cryptosporidiosis was a risk factor for G. duodenalis , E. bieneusi , and C. difficile  infection. In addition, a lower diversity of G. duodenalis, E. bieneusi, and C. difficile genotypes/subtypes was observed in Ward A.The occurrence of Data from this study suggest that multiple pathogens were concurrently present during the previous cryptosporidiosis outbreak. Examination of multiple enteric pathogens should be conducted when poor hygiene is the likely cause of outbreaks of diarrhea. Giardia duodenalis, Enterocytozoon bieneusi, and Clostridium difficile is poorly understood in developing countries despite their wide occurrence. Because they are transmitted by the same fecal-oral route as Cryptosporidium, in this study, we have examined the occurrence of these enteric pathogens in children during a cryptosporidiosis outbreak in a pediatric hospital in China. Using molecular diagnostic tools, we have detected significantly higher infection rates of these enteric pathogens in the outbreak ward than in two control wards in the same hospital. We have also shown a much higher occurrence of these pathogens in children having cryptosporidiosis than those having no cryptosporidiosis. We have demonstrated that the genetic diversity of enteric pathogens is much lower in the outbreak ward than in control wards. Therefore, other enteric pathogens are concurrently present during the cryptosporidiosis outbreak, and examinations for multiple enteric pathogens should be conducted when poor hygiene is considered the likely cause of outbreaks of diarrhea.The transmission of  Cryptosporidium is a significant cause of diarrhea in humans worldwide Cryptosporidium infections through the fecal-oral route via direct person-to-person or animal-to-person contact, or ingestion of contaminated water or food Cryptosporidium, pathogens like Giardia duodenalis, Enterocytozoon bieneusi, and Clostridium difficile are also significant causes of diarrhea in humans worldwide and can be transmitted from persons to persons by the same fecal-oral route involved in cryptosporidiosis occurrence Clostridium difficileSimilar to Cryptosporidium in in-patients from three pediatric hospitals, P. R. China, we identified an extended outbreak of cryptosporidiosis in a pediatric hospital ward , with more than 50% (38/74) children affected by two C. hominis subtypes (IaA14R4 and IdA19) during a 14-month period (Sep. 2007–Oct. 2009) Cryptosporidium species and C. hominis subtypes were significantly lower in Ward A than in other wards/hospitals, with only one species (C. hominis) and two C. hominis subtypes (IaA14R4 and IdA19) being found in 38 patients in Ward A while four species of Cryptosporidium and six C. hominis subtypes being found in 62 patients in other wards and hospitals In one recent molecular epidemiologic study on G. duodenalis, E. bieneusi, and C. difficile in hospitalized children in Ward A with those in two control wards in the same hospital: Ward C for patients having hemophilia, anemia, and neurological diseases, and Ward D for patients having general surgeries. This was the first study to use genotyping and subtyping tools to investigate the transmission of multiple enteric pathogens during a cryptosporidiosis outbreak.Because concurrent infections of multiple pathogens are sometimes involved in gastroenteritis in hospitalized children Written informed consent was obtained from the parents or guardians of the children. This study was approved by the Ethics Committee of the East China University of Science and Technology.All specimens for this study were collected from in-hospital children during September 2007–October 2009 as described Cryptosporidium infection rate = 51.4%), where the cryptosporidiosis outbreak occurred, was regarded as the case ward, while two other wards  in the same hospital  without cryptosporidiosis outbreak were regarded as the control wards. Overall, 573 children, including 74 from Ward A , 283 from Ward C , and 216 from Ward D , were examined for the occurrence and genotype/subtype distribution of G. duodenalis, E. bieneusi, and C. difficile. In addition, 2,672 children from other known or unknown wards in Hospital I , 489 children from Hospital II , and 311 children from Hospital III  in the same city, were also examined for G. duodenalis. Information on age, gender, and the occurrence of diarrhea as defined by the attending physicians was collected for each patient as previously described In this study, Ward A  gene was amplified by nested PCR Giardia (bg) and a 530-bp fragment of the glutamate dehydrogenase (gdh) gene were further amplified from DNA of the tpi-positive specimens Giardia duodenalis genotypes and subtypes were determined using the established nomenclature system based on multilocus sequence data Genomic DNA was extracted from 0.2 ml of fecal materials using a FastDNA SPIN Kit for Soil . To detect rRNA gene containing the entire internal transcribed spacer (ITS) was amplified and sequenced to detect and identify E. bieneusi genotypes E. bieneusi were named according to established nomenclature tcdB gene was used to detect C. difficileClostridium difficile in tcdB-positive specimens was subtyped by sequence analysis of the slpA gene as previously described A ∼392-bp fragment of the http://technelysium.com.au/?page_id=27). The accuracy of the sequencing reads was confirmed by bidirectional sequencing. The nucleotide sequences of G. duodenalis, E. bieneusi, and C. difficile genotypes/subtypes obtained were aligned with reference sequences of each genetic locus downloaded from GenBank using ClustalX . A neighbor-joining analysis of the aligned sequences was performed with the program Mega 5 (http://www.megasoftware.net/). Unique nucleotide sequences generated from the study were deposited in GenBank under accession numbers JX994231-JX994292.All positive PCR products generated in the study were directly sequenced using Big Dye Terminator v3.1 Cycle Sequencing Kits  and an ABI 3130 Genetic Analyzer (Applied Biosystems). Sequences were assembled using ChromasPro (version 1.5) software . Differences were considered significant at P≤0.05. All statistical analyses were performed using the SPSS Statistics 17.0 .The χCryptosporidium, G. duodenalis, E. bieneusi and C. difficile were analyzed in this study and no examinations of bacteria or viruses were conducted. The Cryptosporidium infection rates were 51.4% (38/74) in case Ward A while 1.8% (5/283) and 2.3% (5/216) in control Wards C and D, respectively G. duodenalis at the tpi locus  and control wards (C and D) was significant .Only four parasites including pi locus . In contpi locus . The dif P<0.01; . In addiE. bieneusi at the ITS locus, with eight positives in each ward. The infection rate of E. bieneusi in Ward A (10.8%) was significantly higher than those in Ward C (2.8%) and D (3.7%) , 107/283 (37.8%), and 60/216 (27.8%) in Wards A, C, and D being positive, respectively  were positive for ectively . The inf P<0.01; .Cryptosporidium, G. duodenalis, E. bieneusi, and C. difficile, were detected in both the case and control wards. Comparing with the control Wards C and D, Ward A had a significantly higher overall infection rate of enteric pathogens . Over half of children with enteric pathogens in Ward A (31/58 or 53.4%) were concurrently infected with two or more pathogens, while only a small number of children with enteric pathogens in Ward C (4/119 or 3.4%) or D  were infected with multiple pathogens (P<0.01).Concurrent infections of multiple pathogens, including Cryptosporidium-positive children had higher infection rates of G. duodenalis , E. bieneusi , and C. difficile  than Cryptosporidium-negative children . In contrast, children ≤6 months were more likely infected with E. bieneusi , and children ≤12 months were more likely infected with C. difficile . In contrast, in children older than 12 months, only G. duodenalis was significantly more prevalent in Ward A than in the controls . The other three includes EbpC (1 case), EbpA (2 cases), and D (1 case). Twelve novel genotypes (SH1–12) were found in this study, with SH2 in three cases and all other genotypes in one case each  than in Wards C and D , compared to kr-03 in control Wards C (23/74 slp-A positive cases) and D . Infection with Cryptosporidium was significantly associated with the occurrence of diarrhea . However, a large number of asymptomatic G. duodenalis, E. bieneusi, and C. difficile infections were observed in both case and control wards in this study. In addition, in Ward A, the difference in diarrhea rates between children with multiple infections and children with single infection was not significant . This was also the case in the control Wards C and D .A significantly higher diarrhea rate was observed in Ward A than in control Wards C and D , Hospitals II (0.6%), and III (1.6%) are similar to those reported in out-patients and inpatients (0.2–0.6%) and the general population (2.5%) in China E. bieneusi in hospitalized children in Wards C (2.8%) and D (3.7%) are also very low. In contrast, significantly higher infection rates of G. duodenalis  and E. bieneusi  were seen in Ward A, indicating that these pathogens were transmitted frequently within this ward during the cryptosporidiosis outbreak. Although a high carriage of C. difficile was found in control Wards C (37.8%) and D (27.8%), this is similar to the 16–35% carriage of C. difficile in hospital inpatients in other countries C. difficile in Ward A . Previously, no data existed on the prevalence of C. difficile in hospitalized children in China, although an infection rate of 9.5% was reported in adults in Shanghai The low infection rates of G. duodenalis, E. bieneusi, and C. difficile found in Ward A versus in control wards also supports the hypothesis of concurrent transmission of these enteric pathogens during the cryptosporidiosis outbreak. For G. duodenalis, AII was the only subtype seen in Ward A, although assemblage B and both AI and AII subtypes of assemblage A are commonly found in humans around the world (7), and they were all found in other wards and hospitals in the present study . Of note is the significant association of the three enteric pathogens examined in this study and occurrences of cryptosporidiosis in these children , whereas the dominant subtype kr-03 in control wards was toxin A-positive and toxin B-positive (A+B+) C. difficile subtypes and the occurrence of diarrhea, although we previously showed a link between cryptosporidiosis and diarrhea in these children slpA subtypes including sh-01 and sh-02 were found in many children from all three wards  and low occurrence of mixed infections in control Wards C and D (9 cases of mixed infections versus 178 cases of single-pathogen infection). In addition, with the exception of Cryptosporidium, none of the other pathogens examined in this study were among the recently identified major pathogens for moderate-to-severe diarrhea in the Global Enteric Multicenter Study The results of this study and our previous study Cryptosporidium-positive children were co-infected with G. duodenalis, E. bieneusi, or C. difficile. The young age of affected children and concurrent infections with multiple enteric pathogens clearly implicated poor diaper changing and hand washing by hired caregivers as the cause of the outbreak. Thus, better training of caregivers on hygienic practices such as hand washing and proper use of disposable gloves and disinfectants is needed to reduce the risk of pathogen transmission in healthcare facilities. In conclusion, using genotyping and subtyping tools we retrospectively identified a multi-pathogen outbreak in a pediatric hospital ward. As reported previously


Alcohol problems are a serious public health concern, and few problem drinkers ever seek treatment. The Internet is one means of promoting access to care, but more research is needed to test the best types of interventions to employ. Evaluation of Internet-based interventions that contain a variety of research-validated cognitive-behavioral tools, which have been shown to be helpful to those with more severe alcohol concerns, should be a priority.To evaluate whether providing access to an extended Internet intervention for alcohol problems offers additional benefits in promoting reductions in alcohol consumption compared with a brief Internet intervention. The hypothesis for the current trial was that respondents who were provided with access to an extended Internet intervention (the Alcohol Help Center [AHC]) would display significantly improved drinking outcomes at 6-month follow-up, compared with respondents who were provided with access to a brief Internet intervention (the Check Your Drinking [CYD] screener).A single-blinded randomized controlled trial with a 6-month follow-up. A general population sample of problem drinkers was recruited through newspaper advertisements in a large metropolitan city. Baseline and follow-up data were collected by postal mail.P = .046).A volunteer sample of problem drinkers of legal drinking age with home access to the Internet were recruited for the trial. Of 239 potential respondents recruited in 2010, 170 met inclusion criteria . Follow-up rates were 90.0% (153/170) with no adverse effects of the interventions reported. A repeated-measures multivariate analysis of variance of the outcome measures using an intent-to-treat approach found a significantly greater reduction in amount of drinking among participants provided access to the AHC than among participants provided access to the CYD  Alcohol is the third-leading cause of preventable death . UnfortuInternet-based interventions have been identified as one promising option. Several reviews have concluded that there is a fast-developing evidence base for the efficacy of these interventions -8, partiIn this paper we report results of a randomized controlled trial testing the added benefit of providing access to such an extended Internet-based intervention, which contained an extensive array of cognitive-behavioral tools for problem drinkers (the Alcohol Help Center [AHC]), over the provision of a brief, personalized feedback Internet-based intervention (the Check Your Drinking [CYD] screener). Both of these Internet-based interventions are available free of charge on the Internet, making evaluations of their use of immediate benefit to problem drinkers. In addition, the CYD has already been subjected to four randomized controlled trials, in which the provision of this brief intervention yielded reductions in alcohol consumption among participants in a variety of different settings -12. In tParticipants were recruited through newspaper advertisements in metropolitan Toronto, Canada (May to September 2010) asking for current drinkers interested in helping “in the process of developing and evaluating Internet-based interventions for alcohol users.” Interested potential participants called the telephone number provided and left their name and address to be sent a consent form and a baseline questionnaire by postal mail. The questionnaire contained a graphic describing a standard drink .Those returning the signed consent form and the completed baseline questionnaire were randomly assigned into one of two conditions: to be provided access to the brief Internet-based CYD brief personalized feedback intervention or to be provided access to the extended Internet-based AHC. Access was provided by sending each participant a unique password to be entered into a study-specific online portal . Random numbers where generated in blocks of two using an online random numbers generator  by the author. The random numbers list was used sequentially by a research assistant, who sent out the relevant intervention letter to participants in the order that their consent forms were returned. Participants were blind to the different experimental conditions in the study. Participants were followed up at 6 months and were compensated Can $20 for completing the follow-up questionnaire . Follow-ups were conducted from November 2010 to June 2011. If participants did not return their original follow-up questionnaire within 1 month, they were sent a second questionnaire by postal mail. The research assistant in charge of follow-up was not blind to experimental condition, as the follow-up questionnaires were slightly different . The trial was approved by the standing research ethics committee for the Centre for Addiction and Mental Health.As per protocol, the primary outcome measures were number of drinks consumed in a typical week, highest number of drinks on one occasion in the last 6 months, frequency of drinking, typical quantity of drinking on one occasion, and frequency of drinking 5 or more drinks on one occasion . These last 3 items were combined to form the Alcohol Use Disorders Identification Test-Consumption subscale (AUDIT-C) . For parResults from a related self-help trial  were useThe CYD screener (CheckYourDrinking.net) is a brief and personalized 18-item screener that has been designed to assess quantity and frequency of drinking, and the severity of drinking problems . FollowiDrinkWise [Alcohol and You, developed by the principal investigator of this study [The AHC (AlcoholHelpCenter.net) is a website that has been developed to contain the cognitive-behavioral, motivational, and relapse prevention components that have been found to be effective in well-validated self-help books and other brief interventions. Specific sources for the content include (1) components found in the self-help book rinkWise , (2) exeis study , and (3)is study . The maiis study , (2) an P > .05). Participants’ mean age was 45.2 (SD 12.2), 59.4% (101/170) were male, 58.8% (100/170) had some postsecondary education, 39% (67/170) were married or living with a partner, 55% (94/170) were employed full- or part-time, and more than half  had a yearly household income of Can $30,000 or more. Baseline drinking was heavy, with participants reporting an average AUDIT score of 22.1 (SD 7.6), typical weekly consumption of 31.7 (SD 18.4) drinks, and highest number of drinks consumed on one occasion of 13.7 (SD 7.2). A total of 71.8% (122/170) of participants used the password and logged in to their respective online intervention; however, all participants were included in the analysis in an intent-to-treat approach. Follow-up rates were good, with 90% (n = 153) completing the 6-month follow-up. There were no significant differences (P > .05) between experimental conditions, demographic characteristics, or drinking characteristics in the proportion of participants who completed the 6-month follow-up.A total of 170 participants met inclusion criteria for this trial. Bivariate comparisons compared baseline demographic and drinking characteristics between experimental conditions  and found no significant differences between them . In addition, there was a significant interaction between time and condition . Further univariate tests found that the time × condition interaction was not significant for the number of drinks in a typical week variable , approached significance for the AUDIT-C variable , and was significant for the highest number of drinks on one occasion variable . A multivariate analysis of variance was conducted and found a main effect for time  toward an additional benefit of the AHC over the CYD on the AUDIT-C scores, a scale that is a composite measure of hazardous drinking. Finally, participants in both groups reduced their drinking in a typical week substantially (by 4.9 drinks per week in the CYD condition and 6.3 drinks per week in the AHC condition). These reductions are similar in scope to that seen in an earlier randomized controlled trial comparing the CYD with a no-intervention control [The current trial found a small but significant additional reduction in drinking among participants who were provided with access to the extended Internet-based intervention (the AHC) as compared with participants receiving the brief Internet-based intervention (the CYD) at a 6-month follow-up. The difference specifically had to do with reductions in amount of alcohol consumed during the participants’ heaviest drinking occasion. Given the increased risks associated with heavy drinking situations , this reA strength, and a simultaneous limitation, of this trial was the severity of current drinking problems in the participants  . Thus, wDespite these limitations, the results of this trial are important, as they demonstrate the added benefit of an extended Internet-based intervention over a brief, personalized feedback intervention for problem drinkers. Systematic replications of these findings would allow the development of an adequate research base supporting this highly accessible and cost-effective means of providing assistance to problem drinkers who might otherwise receive no help for this important health concern.


In the post-Soviet period, Russian working-age men have suffered unusually high mortality rates. Earlier quantitative work found that part of this is attributable to hazardous and harmful patterns of alcohol consumption, which increased in the period of transition at a time of massive social and economic disruption and uncertainty. However, there has been very little work done to document and understand in detail the downward life trajectories of individual men who died prematurely from alcohol-related conditions. Building on an earlier case-control study, this unique qualitative study investigates the perceived interplay between men's drinking careers, their employment and family history, health and eventual death.In-depth interviews were conducted with close relatives (most often the widow) of 19 men who died between 2003 and 2005 aged 25-54 years whose close relatives reported that alcohol contributed to their death. The study was conducted in a typical medium-sized Russian city. The relative's accounts were analysed using thematic content analysis.The accounts describe how hazardous drinking both contributed to serious employment, family and health problems, and was simultaneously used as a coping mechanism to deal with life crises and a decline in social status. The interviews highlighted the importance of the workplace and employment status for shaping men's drinking patterns. Common themes emerged around a culture of drinking in the workplace, peer pressure from colleagues to drink, use of alcohol as remuneration, consuming non-beverage alcohols, Russian-specific drinking patterns, attitudes to treatment, and passive attitudes towards health and drinking.The study provides a unique insight into the personal decline that lies behind the extremely high working-age mortality due to heavy drinking in Russia, and highlights how health status and hazardous drinking are often closely intertwined with economic and social functioning. Descriptions of the development of drinking careers, hazardous drinking patterns and treatment experiences can be used to plan effective interventions relevant in the Russian context. Since the fall of the Soviet Union, mortality rates of Russian men have experienced dramatic fluctuations, diverging considerably from patterns seen in other industrialised countries. Male life expectancy decreased to a low of 57 years in 1994, and despite a modest increase in recent times to approximately 62 years, still lags considerably behind other European nations -9.zapoi. This is a well recognised term in Russia to describe an extended episode of heavy drinking characterised by a period of continuous drunkenness lasting at least two days, where the person withdraws from normal life. He could not stand up. His legs did not support him. It was the first time I called for an ambulance. They said they could stop zapoi if we paid and took him to the narcology dispensary [Russian treatment centre for alcohol and drug problems]." (#2)"zapoi:It was common for the subject not to acknowledge their problem with alcohol and therefore refuse to engage in treatment. Some of the interviewees also denied the fact that the deceased were alcoholics, evidenced by of a lack of More recently he drank continuously but I don't think these were zapois. They say, zapoi is when a person even gets up at night to have a drink. He didn't drink at night and he never missed his work." (#16)"Besides, can we say he was an alcoholic? He didn't have zapois. He got up in the mornings to help me; he went to collect bottles or something... Drank like everyone." (#1)"'zapoi', which was understood slightly differently by each respondent, although it is generally understood to be a period of continuous drunkenness that by its very nature disrupts normal functioning.It is interesting to note the subjective and contested definition of the term zapoi, detoxification programs, and 'coding' (see Background) to prevent consumption for a period of time. No psychological services are offered for free. Private clinics and doctors offer a broader range of services, including counselling, but the cost can be prohibitive.Six of the men sought treatment enabling them to have periods of sobriety and return to normal life and work. Some of the men were unable to recognise their problem; those men that did felt helpless to deal with their dependence; some relatives mentioned that they felt disappointed by the treatment available. In Izhevsk, options for state-funded care is limited to narcology dispensaries or psychiatric departments treating severe cases of alcohol dependence, offering interventions to stop In all cases, employment and drinking history were described as closely intertwined. In general, there were two, almost contradictory, reasons given for starting a heavy drinking career. In the first group, nine of the respondents felt that disruption in the man's workplace and loss of status directly or indirectly triggered heavy drinking. These men were often described as being happy, secure and successful early in their careers, before a 'crisis' situation at work such as redundancy, irregular pay, or conflicts with new management led them to drink more to deal with unhappiness or stress. The loss of stable employment status was identified as traumatic, leading directly to work dissatisfaction, loss of motivation and self-esteem, providing an almost logical reason for starting to drink heavily:"In 1991 [the year of the collapse of the Soviet Union] everything changed. The plant was in a difficult financial situation. They didn't pay less but they had delays in wages. And very often the wages were not paid with money but with the plant's production. He couldn't adapt to such life. He tried to earn money, did two shifts in a row but at the same plant. He worked overtime. It was very difficult. He said he'd lived two lives already. That was the feeling he had. Gradually he drank more and more. Then he began drinking vodka. More recently he drank anything he could get" (#17)Others suffered personal crises causing them to leave work and start drinking:After another such conflict with her mother the girl [the subject's girlfriend] jumped from the window at the fifth floor. He suffered from deep depression. He said: "I don't want to live any more" and quit his second job. After that he didn't work for a year and a half, made friends with alcoholics who drank surrogates."(#9)"On the other hand, in the second group, the remainder of the accounts described a gradual shift into heavy drinking, because they worked in places where alcohol was tolerated and encouraged and where they were pressured to drink with colleagues. Sometimes the men 'settled' for these types of jobs because they fitted in with their drinking habits. Certain professions, such as car mechanic, plumber and carpenter commonly received payment and tips in the form of alcohol rather than money. In these accounts, relatives often blamed the lack of regulation and social pressure in the workplace for the development of alcohol problems:They alcoholise you in the [transport] fleet, I told him. How can I refuse, they ask me to do something with their car, he said. And of course, they give him a bottle, not money for this." (#14)"He went to work and drank....The management didn't pay attention (turned a blind eye) to the fact that the workers drink all the way through the shift. Nobody told them off. They set the machine working for the whole day and just periodically check that it's still going. And so [because of this]-you could do, what you wanted. Once he didn't get back home for three days. I found him in the shop. At nights he had slept there, drunk" (#11)""It all started when he began working there, in that agency... Before that he drank a little sometimes but never drank heavily" (#1)After heavy drinking developed, all describe patterns of subsequent unstable employment and lack of motivation to work, which lasted until death. Periods of employment were broken up with periods of heavy drinking, and ill health, and sometimes these factors were the reason for loss of employment:But a little time after he fell off the wagon [went back to drinking heavily] - perhaps, something happened at work: he began drinking, missed work and was asked to leave... Then he got a job at the radio plant. They sacked him for drinking" (#15)"He didn't work there long. Later he felt worse and worse. Would work a little, get back home and lie down. A few times I told him to go to the doctor. But he answered he wouldn't go anywhere. More recently he was registered at the job centre" (#13)"He didn't have money to buy more vodka that's why he bought Hawthorn" (#13). Furthermore, it was perceived that drinking surrogates subsequently affected men's health, and brought about alcohol dependence, making them less likely to find a job, or more likely to be fired from the next job.The accounts describe how cycles of unemployment, drinking and ill health were mutually reinforcing, steadily bringing a decline in physical and mental health, making death seem inevitable. Rapid organisational changes in the workplace in the post-soviet period left some men unable to adapt, experiencing redundancy or salary delays, leading some of them to drink heavily, (see quotes above) while on the other hand, many were fired from their jobs because of alcohol abuse. Unemployment brought poverty, inducing many to drink surrogate alcohol as a cheaper alternative to beverage alcohol  had been married at some time and most had children. 12 were still married at the time of their death, and the other 6 were divorced or separated, 4 reportedly due to the husband's drinking. It was common for divorced couples to continue living together due to housing pressures.In the majority of cases, men's heavy drinking, frequent He confided in me" (#16) "He understood me. I was able to get from him what I wanted."(#2)), mentioning that he was a good man despite his problems, and were more prone to see the deceased as a victim of circumstance. On the contrary, the divorced men were often blamed for their alcohol problem and subsequent death, ("laziness was the main cause"(#8)), recalled as being a burden to their families, and family relations were reported as non-existent or conflict-ridden.Widows married at death tended to report good relations of mutual love, support and understanding "We treated him the way he treated us. He would run after us with an axe, beat us badly, shout." (Son of deceased (#6))zapoi, how this may have influenced the man was not discussed. Two of the widows mentioned that they drank together with their husbands.Seven of the 19 interviews reported that the men had parents or siblings who were heavy drinkers or who went on In the accounts very heavy drinkers, especially surrogate drinkers, were stigmatised and became socially disconnected, either drinking in isolation, or spending time only with problem drinkers. The man's 'real' friends, perhaps from childhood, tended to drift away and to be replaced with a circle of drinkers. Drinking companions, although supportive and loyal in their own way, were a bad influence, encouraging him to drink surrogates, drink more regularly, and to borrow money.The most common approach towards excessive drinking was for the spouse/relative to try to control his consumption, by persuading him to drink at home, rather than with colleagues or in the street, refusing to give money for alcohol, pouring away alcohol, and setting limits to the man's drunkenness in exchange for money, although these strategies had little effect.On the other hand, many of the widow's accounts describe what might be labelled in the addiction field as 'co-dependent' and 'enabling' behaviour and attitudes, comprisi"We never had arguments - I always tried to avoid them....Never criticized him for drinking. Always tried to support him. When he was sober he was a very good man" (#15),And some provided 'safe' home environments for drinking:"The only thing I taught him not to do was not to drink in the street. You'd better come and drink at home to avoid problems, I told him. (#16)so that less remained for him". (#1) (2 cases), protecting men from the consequences of their drinking by providing excuses at their workplace (2 cases), and helping them to find another job after they had been sacked for drinking (3 cases). Relatives tended to be sympathetic and supportive if they were also concordant drinkers, or if they felt that alcohol was alleviating suffering:Obvious examples of enabling, facilitative behaviour are widows providing money for strong alcohol and surrogates (9 of the proxies), drinking together with him "When he was unable to walk any longer we bought it for him because he needed it. Otherwise he would cry without stopping. We diluted it well (a bottle for 1 or 1.5 litre) and left it next to his bed. It was enough for a day. Sometimes he forgot he'd already drank it. He'd drink and ask for it again. When he drank it he calmed down and had a sleep. More recently he would forget everything. He didn't eat anything during the last week. He only drank. He grew yellow." (#15)"A few acknowledged that excessive tolerance on behalf of women could make them partly responsible for the problem:Perhaps, we, the women, make them like this, we try to solve their problems, we nurse them" (#15)""I think to some extent mum is to blame for it too...when he's got a hangover mum carries fried eggs with gherkins to him. "Why, Mishen'ka [pet name for Mikhail] is suffering"(#15)I didn't insist on coding because I think coding is the end of the line.(#7)"), describing alcoholism as an illness caused by innate weakness or a product of social forces.Most thought that treatment was pointless if the man didn't admit he had a problem, and a few rejected the idea of treatment completely, especially coding "Another also died young. I don't know if it was related to drinking or not. We know it's poison but what can we do" (#1)Some felt that heavier drinking was a result of post-Soviet changes: previously the Soviet state had provided more structured activities for schoolchildren and young people, job and wage security for adults which prevented problem drinking in men and young people. Young people and men were described as responding passively to alcohol advertising and difficult structural conditions, implying that state intervention and education were needed to solve the problem. Men were described as innately weaker than women, unable to resist invitations to drink, suffering from social stress more readily, lacking in ambition and passive in the face of health problems. By contrast, women's domestic responsibilities gave them strength and purposeThe deceased were commonly described as being caught in a downward spiral of heavy drinking, ill health, work and personal problems, where life problems were a motivation for heavier drinking, and heavier drinking, in turn, worsened the man's situation. In many cases, this vicious cycle meant that early death was perceived as inevitable. The mutual reinforcement between life crises and heavy drinking makes it difficult to determine generalisable causal relationships. Instead, this paper focuses on in-depth, detailed accounts to illuminate and contextualise the poor mortality statistics found in post-soviet Russia.The employment context was understood to play a crucial role in the formation of men's drinking habits, with both drinking leading to unemployment, and work problems leading to heavy drinking. The men's biographies confirm that the relationship between heavy drinking and economic/social problems frequently runs in both directions, and in most cases it is difficult to separate them. Unstable employment was the most commonly cited explanation for starting a heavy drinking career. This could reflect the widely held idea that profession and stable employment is crucial for a man's sense of identity and self-esteem, and instability in this arena is understandable cause for drinking ,27.zapois began in the workplace or with male colleagues, supporting previous research which suggested that the workplace is a very common, acceptable drinking context in Russia, much more so than in other countries [However at the same time most of the men's heavy drinking and ountries . Some inountries  is relatzapois, and 'heavy drinking' when the man lost control, and was labelled an 'alcoholic'. 'Alcoholics' were strongly disapproved of and stigmatised, and alcohol abuse was seen as a social, rather than health problem. There was little perception of a stage in the man's career when treatment might have been effective. More work could be done to build public awareness of how alcohol problems develop and what can be done to treat them. In addition, some of the accounts complained that state-run services were inadequate. One relative stated that a narcology clinic refused to provide treatment as the man was a repeat patient, and others complained about poor organisation and lack of referrals. Some of the respondents were sceptical about the effectiveness of 'coding' treatment.Beliefs that alcoholism is untreatable and alcoholics are beyond help meant that those suffering with alcohol problems, and their close relatives, sometimes do not try to seek treatment. In the descriptions of drinking careers, there was a sharp contrast between 'normal' everyday drinking, which was often tolerated, approved of and might include occasional It was striking that virtually all of the respondents described the men as taking a passive, helpless stance to alcohol, alcohol treatment, and general health. They often refused to go the doctors, even when seriously ill, take prescribed medication, take advice from narcologists, or attempt to reduce their alcohol intake by themselves. Accounts of passivity could be a technique of remembrance for the relative, helping them to explain the death and assign responsibility. Reluctance to seek help could also reflect dissatisfaction and disillusionment with available treatment options for alcoholism. But the findings are in line with other studies arguing that post-Soviet Russian culture does not give a central place to people taking personal responsibility for their own health. Instead there is a persistence of an outlook fostered in the Soviet period which emphasised paternal protection and authority of medical personnel,37. OtheExplanations for excessive drinking and death varied according to the respondent's structural and emotional relationship to the deceased. Children, siblings, and divorced widows more readily attributed personal blame to the man for losing employment, drinking heavily and refusing treatment, whereas those still married emphasised his victim status and his helplessness in the face of difficult external conditions. This contrasted with more uniform 'lay' explanations found elsewhere.There are some methodological limitations to the study. The purposive sampling technique, in which we selected men whose proxy informant believed that alcohol contributed to the man's death, means that the accounts are likely to depict alcohol as strongly related to employment, health and family relations. For this reason the study cannot throw light on the more general association of alcohol with social factors, as would be seen in a more general sample. The biography approach and order of topics may have encouraged the respondent to construct explanations which are logical, ordered and coherent . The useThis study highlights how, among men that died prematurely of alcohol-related causes, negative life events and heavy drinking are mutually reinforcing, producing a sharp decline in health status. Changes in the workplace connected to post-soviet transformation were frequently cited as reasons for starting heavy drinking. Importantly, however, the accounts revealed that industrial, factory, and manual workplace contexts encouraged, or at least tolerated, a culture of heavy and hazardous drinking that probably extend back well into the Soviet period. These practices may well interact with both socio-economic status and ideas about masculinity, and deserves further exploration. Rather than the 'lay' or generalised perceptions about drinking found in previous qualitative work, this study describes how hazardous drinking was personally tolerated, negotiated, and accepted within couples and families in Russia. The broad range of topics covered in the interviews - hazardous drinking patterns, surrogates, attitudes towards treatment can assist in planning interventions that are salient in the Russian context, and hence most likely to be effective.The authors declare that they have no competing interests.LS designed the study, collected and analysed the data; KK did a secondary analysis of the data and wrote the manuscript; NB helped to draft the manuscript; DL helped design the study and draft the manuscript; DE advised on the analysis and helped draft the manuscript. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2458/11/481/prepub


This is the second Thematic Series in the Beilstein Journal of Organic Chemistry focused on natural products. While the first Thematic Series summarized recent research work on the biosynthesis and function of natural products –2, the pWhy the research interest in natural products? Natural products are structurally fascinating as is spectacularly demonstrated by the prominent examples of maitotoxin , the larArtemisia annua, which is highly efficient in the treatment of malaria. The problem is that the plant produces only small and variable amounts, thereby making the isolation procedure difficult and expensive. Neither biotechnology nor chemistry were able to provide a solution on their own, but the combination of the heterologous production of an advanced intermediate in yeast [Why should we have a look at the synthetic and biosynthetic aspects of natural products in this Thematic Series? Of course, the chemistry of nature and as it is used by chemists is one and the same and makes use of the same intrinsic reactivity of molecules. It is fascinating to see that a particular chemical reaction, believed to be invented by man, is in fact already used for millions – sometimes even billions – of years by Mother Nature. An interesting example is the discovery of the Diels–Alder reaction in the 1920s and its later enhancement into an enantioselective reaction by the development of chiral catalysts. But a Diels–Alder reaction also occurs in the biosynthesis of lovastatin and is likely catalysed by a Diels–Alderase , a naturin yeast  and its in yeast  procuresI would like to thank the highly professional team of the Beilstein-Institut for a very pleasant cooperation on this Thematic Series. I wish you, the reader, joy and fun with this Thematic Series and – at the best – a new impetus for your own future research!Jeroen S. DickschatBraunschweig, September 2013


Client change talk has been proposed as a mechanism of change in motivational interviewing (MI) by mediating the link between therapist MI-consistent behaviors (MICO) and client behavioral outcomes. In the present study, we tested under what circumstances this generic MI conceptualization was supported. The study context was a Swiss clinical trial of brief MI for heavy drinking among non-treatment seeking young men (age 20). We conducted psycholinguistic coding of 174 sessions using the MI Skill Code  and derived the frequency of MICO and the strength of change talk (CTS) averaged over the session. CTS was examined as a mediator of the relationship between MICO and a drinking composite score  measured 3 months after baseline, controlling for the baseline drinking composite score. Finally, we tested therapist gender and MI experience and client readiness to change and alcohol problems severity as moderators of this mediation model (see Hayes 2013). CTS significantly predicted outcome in the expected direction (i.e. higher strength related to less drinking), but MICO did not significantly predicted CTS. However, CTS mediated the relationship between MICO and drinking outcomes when therapists had more experience in MI and when clients had more severe alcohol problems . Findings showed that the mechanism of change hypothesized by MI theory was operative in our brief MI model  with heavy drinking young men, but only under particular conditions. Our results suggest that particular attention should be paid to therapist selection and/or training and supervision until they reach a certain level of competence, and that MI might not be appropriate for non-treatment seeking clients drinking at lower level of risk.


AbstractTrachymolgus purpureus Fisher & Dowling sp. n. is described from the Ozark highlands of North America. A diversity of imaging techniques are used to illustrate the species including low-temperature scanning electron microscopy (LT-SEM), stereomicrography, compound light micrography, and digitally created line drawings. Developmental stages  and morphology are illustrated and discussed, and terminological corrections are suggested. Trachymolgus recki Gomelauri, 1961 is regarded as being described from tritonymphs. A key to Trachymolgus is presented. Bdellidae Dugès, 1834 generally have a striated, unsclerotized integument. Exceptions occur in Cytinae Grandjean, 1938, which comprises three of the most distinctive bdelloid genera. Cyta Heyden, 1826 are common mites known for their stocky bodies, massive chelicerae, and unpaired fifth eye. Rigibdella ignea Tseng, 1978 from Taiwan have sclerotized, striated holodorsal shields  was originally collected in northern Italy (Padova) and subsequently reported from Sicily and Lugano, Switzerland (Cyta and Trachymolgus into a new subfamily (Cytinae) based on the number of subcapitular setae and leg trichobothria, presence of pseudotracheae, and normal chelicerae (reduced in Spinibdellinae). Two other species were described later, both known only from their type localities: Trachymolgus recki Gomelauri, 1961 in Georgia (former U.S.S.R.: Tbilisi and Kashtak) and Trachymolgus jesusi Mejia-Recamier & Palacios-Vargas, 1999 in Mexico . Undetermined species have been reported from a Buddhist temple on Shikoku Island, Japan , specifically Buffalo National River and Devil’s Den State Park, and extracted using Berlese-Tullgren funnels. Approximately half of the specimens were slide-mounted with Hoyer’s medium (see vs) and dorsal subcapitular setae (ds).An effort is made to implement terminology that is broadly applicable and well accepted across acariforms despite conventions used among bdelloid authors. Thus, two terms have been renamed herein. First, “hypostome” is used by many  to referBdellidae: Cunaxidae: Bdelloidea. Prodorsum is widely used by acarologists, but is usually poorly defined and represents the opposite extreme from aspidosoma--straying too far from morphological hypotheses. An exception to this is in Oribatida  and internal/external scapulars (sci and sce). In Bdelloidea, sci are always external to sce, leaving the internal/external designations unintuitive. sce and sci are descriptive, which they have published since , and lateral and median proterosomal setae (lps and mps).With regard to hysterosomal setal notation, we follow the chaetotaxic system of , 1947 thed since . As a reEupodina, and only distal tarsal setae are denoted presently, which has been adopted by other eupodine authors . Nevertheless, we believe Grandjean’s system can be employed with other leg setae, and will readdress this in a more detailed forthcoming study.Leg chaetotaxy follows Grandjean’s system as reviewed by Norton (1977). However, leg chaetotaxy is poorly studied among Most species descriptions include only a few image types; line drawings are most common in acarology e.g., . We beliLine drawings were created digitally with Adobe Illustrator CS5 and a Wacom Cintiq 21UX tablet using procedures outlined in PageBreak flat specimen holders consisting of 16x30mm copper plates that were tacked on the edges to the tabs with a small dot of Tissue Tek , which acted as the cyro-adhesive upon freezing. The samples were frozen conductively, in a Styrofoam box, by placing the plates on the surface of a pre-cooled (-96°C) brass bar whose lower half was submerged in liquid nitrogen (LN2). After 20–30s, the holders containing the frozen samples were transferred to a LN2 Dewar for future use or cryotransferred under vacuum to the cold stage in the pre-chamber of the cryotransfer system. Removal of any surface contamination (condensed water vapor) took place in the cryotransfer system by etching the frozen specimens for 10–15 min by raising the temperature of the stage to -90°C. Following etching, the temperature was lowered below -130°C, and a magnetron sputter head equipped with a platinum target, was used to coat the specimens with a very fine layer of platinum. The specimens were transferred to a pre-cooled (-130°C) cryostage in the SEM for observation. An accelerating voltage of 5kV was used to view the specimens. Images were captured using a 4pi Analysis system . Images were sized and placed together into figures using Adobe® Photoshop 7.0 and CS4.Low-temperature scanning electron micrographs (LT-SEM) were made using an S-4700 field emission scanning electron microscope  equipped with a Quorum CryoPrep PP2000  cryotransfer system. To prepare specimens, mites were placed on 12 mm diameter ultra smooth carbon double sided adhesive tabs  which were adhered toFisher & Dowlingsp. n.urn:lsid:zoobank.org:act:E0FAE922-2B81-4FB5-8D50-CE0F93517CD2http://species-id.net/wiki/Trachymolgus_purpureusTrachymolgus purpureus sp. n. is heavily armored with distinctive integument characteristic of Trachymolgus  and a serrated edge proximal to the tooth (undescribed in other species). All stages have two pairs of eyes, unlike the larva, proto- and deutonymphs of Trachymolgus jesusi, which lack eyes (tritonymphs and adults have two pairs). Trachymolgus purpureus pedipalpal basi- and telofemora are only fused dorsally. Trachymolgus jesusi pedipalp femora are completely fused, whereas Trachymolgus nigerrimus are completely divided. Trachymolgus purpureus, like other Trachymolgus,have undivided femora on legs I-II (femora III-IV are divided). All other Bdellidae have divided femora on all legs. Trachymolgus jesusi is the only bdellid reported to have undivided femora on legs II and III. The ontogeny of Trachymolgus purpureus differs markedly from that described for Trachymolgus jesusi, the only other species where ontogeny was investigated. Finally, there are many chaetotaxic differences on the appendages and venter between Trachymolgus purpureus and Trachymolgus jesusi. Most chaetotaxy of Trachymolgus nigerrimus remain to be investigated. See Remarks for discussion of Trachymolgus recki.hymolgus –2. Like PageBreakgreen coloration, which rendered the normally purple mite teal; teal specimens returned to purple after a few days in 95% ethanol, and were indistinguishable from normal specimens when slide-mounted . Integument divided into heavily armored sclerites with foveolate sculpturing . Dorsal membrane  striated and accompanied with raised bumps similar in size to the foveolate indentions  covering the stigmata. Two pairs of eyes present. Two pairs of minutely barbulate trichobothria: anterior (at) and posterior trichobothria (pt). Barbules are difficult to discern with light microscopy (lps) and median proterosomal setae (mps). Setae lps are oriented dorsomedially and lay in a groove posterior to the first pair of eyes  and seven barbulate setae as follows: c1, c2, d1, e1, f1, f2, and h1. Posteriorly, the hysterosomal shield folds inward between the f1-2 and h1 forming a curved lateral furrow isolating h1 on a raised area.Dorsal idiosoma. Idiosomdentions , 5a-b. Pcroscopy . Two pai of eyes ; mps areapparent  that we Lateral shieldseach posteriorly containing one lyrifissure (ih) and one barbulate seta (h2). Podocephalic canals lead from the posteriolateral edges of the gnathosoma and curve around coxal field I, and are visible externally . Legs(p) and unguinals (u). Proximally, the dorsal setae are as follows: iterals (it), tectals (tc), and fastigials (ft). The tectals are paired on all legs except IV; PageBreakfastidials are paired only on leg I, and are absent on leg IV. Other setal homologies remain to be investigated. Baculiform solenidia present on genua I-IV (σ), tibiae I and III (γ), and tarsi I, II, and IV (ω); short, ceratiform solenidia present on tibiae I and II (γ); and a short solenidion present on tarsi I that has the appearance of being raggedly broken, interpreted here as the famulus (ε). Trichobothria present on tibiae I and IV, and tarsus III. Apotele with barbulate ungues and pulvilli with tenant hairs . Legs, 7: coxaly). Legs. Trochanly). Legs, 9a; othly). Legs. Podomernt hairs . Leg artSubcapitulum (ad), one pair of smooth anterior setae (avs), and one pair of barbulate posterior setae (pvs); dorsally with one pair of smooth, thin, straight setae (ds) that are hidden under the chelicerae in life; ending in three pairs of lateral lips  finely barbulate . John Kethley recollected three specimens from the same bluff a few years later. Another specimen  was collected by Evert Lindquist in the St. Lawrence Islands National Park, Canada . Morphologically, these specimens cannot yet be distinguished from Trachymolgus purpureus, potentially extending the range to eastern U.S. We were able to extract DNA from one of these specimens and will publish our findings later.In the early 1980s, , Canada , but theTrachymolgus is Mexico (Trachymolgus jesusi), Ozark highlands (Trachymolgus purpureus), central Ohio (undet. species), and the northern Appalachian mountains (undet. species). Other groups have a similar distribution, and the biogeographic affinity between the Ozark and Appalachian mountains, and between Mexico and the eastern U.S. has been well documented. Examples include mosses , and were collected near the surface during the winter. When preparing live specimens for LT-SEM, mites are set atop a metal bar that is subjected to liquid nitrogen fumes which freezes them mid-stride for imaging. When Trachymolgus purpureus was subjected to liquid nitrogen temperatures however, they would simply run, curl their legs, and roll off the plate , 23 Sep 2009, by JR Fisher & MJ Skvarla, APGD 09-0923-006.. HOLOTYPE: female, collected from leaf litter, USA, Arkansas, Washington Co., Devil’s Den State Park : 2 individuals collected from leaf litter on rocky slope, USA, Arkansas, Washington Co., Devil’s Den State Park , 28 Aug 2008, by APG Dowling, APGD 08-0828-004 ● 2 individuals collected from leaf litter on rocky slope, USA, Arkansas, Washington Co. Devil’s Den State Park , 30 Aug 2009 by JR Fisher, APGD 09-0830-001 ● 1 individual collected from leaf litter, USA, Arkansas, Newton Co., Buffalo National River, Roark Bluff , 7 Sep 2009 by JR Fisher, APGD 09-0907-005 ● 1 individual collected from American beech leaf litter, USA, Arkansas, Newton Co., Buffalo National River, Boen Gulf , 14 Mar 2010 by JR Fisher, APGD 10-0314-019 ● 1 individual collected from litter on rocky bluff, USA, Arkansas, Newton Co., Buffalo National River, Roark Bluff , [date unknown] by Cal Welbourn, OSAL 0061853. Male (n=5): 1 individual collected from rocky overhang, USA, Arkansas, Washington Co., Devil’s Den State Park  by APG Dowling, APGD 08-0822-001 ● 1 individual collected from cedar litter, USA, Arkansas, Washington Co., Buffalo National River, Roark Bluff  by JR Fisher, APGD 09-0802-006 ● 1 individual collected from leaf litter on rocky slope, USA, Arkansas, Washington Co. Devil’s Den State Park , 30 Aug 2009 by JR Fisher, APGD 09-0830-001 ● 1 individual collected from leaf litter on rocky slope, USA, Arkansas, Washington Co. Devil’s Den State Park , 30 Aug 2009 by JR Fisher, APGD 09-0830-003 ● 1 individual collected from leaf litter, USA, Arkansas, Newton Co., Buffalo National River, Roark Bluff , 7 Sep 2009 by JR Fisher, APGD 09-0907-005. 1 individual collected from leaf litter, USA, Arkansas, Newton Co., Buffalo National River, Boen Gulf , 10 Apr 2010 by APG Dowling. Tritonymph(n=6): 3 individuals collected from cedar litter, USA, Arkansas, Washington Co., Buffalo National River, Roark Bluff  by JR Fisher, APGD 09-0802-006 ● 1 individual collected from oak litter, USA, Arkansas, Washington Co., Buffalo National River, Steel Creek trail  by JR Fisher, APGD PageBreak09-0802-001 ● 1 individual collected from leaf litter, USA, Arkansas, Newton Co., Buffalo National River, Roark Bluff , 7 Sep 2009 by JR Fisher, APGD 09-0907-005 ● 1 individual collected from litter on rocky bluff, USA, Arkansas, Newton Co., Buffalo National River, Roark Bluff , [date unknown] by John Kethley, FMNH 2. Deutonymph (n=4): 3 individuals collected from cedar litter, USA, Arkansas, Washington Co., Buffalo National River, Roark Bluff  by JR Fisher, APGD 09-0802-006. 1 individual collected from leaf litter on rocky slope, USA, Arkansas, Washington Co. Devil’s Den State Park , 30 Aug 2009 by JR Fisher, APGD 09-0830-002. Protonymph (n=2): 1 individuals collected from leaf litter on rocky slope, USA, Arkansas, Washington Co., Devil’s Den State Park , 28 Aug 2008, by JR Fisher & MJ Skvarla, APGD 08-0828-004 ● 1 individual collected from cedar litter, USA, Arkansas, Washington Co., Buffalo National River, Roark Bluff  by JR Fisher, APGD 09-0802-006. Larva (n=1): 1 individual collected from oak litter, USA, Arkansas, Washington Co., Buffalo National River, Steel Creek trail  by JR Fisher, APGD 09-0802-001.PARATYPES: This species is named for the Latin "purpureus," meaning purple.Trachymolgus recki was described from two specimens from Georgia  and yellowish. There was one tooth on the fixed digit, one pair of eyes, and the pedipalps were inconclusively described as “nearly fused” though they were completely fused in the illustration , 6-6-6-10 , and 8-5-11-10 (Trachymolgus jesusi). Larvae, proto- and deutonymphs of Trachymolgus jesusi were described as having highly reduced eyes. Gomelauri observed only one pair eyes in Trachymolgus recki. This offers significant evidence to suggest the specimens used to describe Trachymolgus recki were immature. Since these specimens were said to have three pairs of genital papillae,PageBreak we suggest the description of Trachymolgus recki was based on tritonymphs. Therefore, Trachymolgus recki is excluded from the key below.We have found that purple  in life . Also, tTrachymolgus jesusi , suggest major deviations from what is known from other Bdellidae. Unfortunately, we were unable to obtain type specimens of this species.Aspects of the morphology and development described for