Novel Approach Identifies SNPs in SLC2A10 and KCNK9 with Evidence for Parent-of-Origin Effect on Body Mass Index

Marja-Liisa Lokki työryhmien Generation Scotland Consortium, LifeLines Cohort Study ja GIANT Consortium jäsenPeer reviewe

A new microtitre plate screening method for evaluating the viability of aerobic respiring bacteria in high surface biofilms

Aims: It is difficult to determine the effects of bactericidal compounds against bacteria in a biofilm because classical procedures for determining cell viability require several working days, multiple complicated steps and are frequently only applicable to cells in suspension. We attempt to develop a compact, inexpensive and versatile system to measure directly the extent of biofilm formation from water systems and to determine the viability of respiring bacteria in high surface biofilms. Methods and Results: It has been reported that the reduction of tetrazolium sodium salts, such as XTT (sodium 3,3′-[1-[(phenylamino)carbonyl]-3,4-tetrazolium]Bis(4-methoxy)-6-nitro) benzene sulfonic acid hydrate), during active bacterial metabolism can be incorporated into a colorimetric method for quantifying cell viability. XTT is reduced to a soluble formazan compound during bacterial aerobic metabolism such that the amount of formazan generated is proportional to the bacterial biomass. Conclusions: We show here, for the first time, that this colorimetric approach can be used to determine the metabolic activity of adherent aerobic bacteria in a biofilm as a measure of cell viability. This technique has been used to estimate viability and proliferation of bacteria in suspension, but this is the first application to microbial communities in a real undisturbed biofilm. Significance and Impact of the Study: This simple new system can be used to evaluate the complex biofilm community without separating the bacteria from their support. Thus, the results obtained by this practice may be more representative of the circumstances in a natural system, opening the possibility to multiple potential applications.Fil: Perez, Leonardo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina. Universidad Politécnica de Catalunya; EspañaFil: Álvarez, B. L.. Universidad Politécnica de Catalunya; España. Universidad de Antioquia; ColombiaFil: Codony, F.. Universidad Politécnica de Catalunya; EspañaFil: Fittipaldi, M.. Universidad Politécnica de Catalunya; EspañaFil: Adrados, B.. Universidad Politécnica de Catalunya; EspañaFil: Peñuela Mesa, Gustavo Antonio. Universidad de Antioquia; ColombiaFil: Morató Farreras, Jordi. Universidad Politécnica de Catalunya; Españ

Comparison of removal efficiency of pathogenic microbes in four types of wastewater treatment systems in Denmark

The aim of the present work was to evaluate and compare the performance in the removal of pathogenic microbes in four different types of decentralized wastewater treatment systems, namely: horizontal flow constructed wetlands (HFCW), vertical flow constructed wetlands (VFCW), biological sand filters (BSF) and biofilters (BF). All the systems analyzed are located in Jutland, Denmark. Water sampling took place during a three months period that covered from winter to spring. Conventional microbial indicators such as Escherichia coli, total coliforms (TC), intestinal enterococci and sulphite-reducing clostridia were quantified using traditional microbiological culture methods, whereas Bacteroides spp. determination was performed by quantitative PCR (qPCR). Other water quality parameters such as dissolved oxygen, biological oxygen demand (BOD5), total suspended solids (TSS), pH, temperature, ammonium concentration and conductivity of influent and effluent water samples were also analyzed. The results showed that bacterial indicators significantly reduced in all the systems analyzed. In general, BF showed the best performance in the removal of microbes for all bacteria studied, while BSF demonstrated an improved capacity to eliminate E. coli and TC. Contrarily, VFCW seems to be more effective reducing the amount of intestinal enterococci, sulphite-reducing clostridia, and Bacteroides spp. In the present study, HFCW were the less efficient wastewater treatment system for the elimination of the evaluated pathogens. However, the performance in the removal of microbes was still significant considering that such systems were the oldest under operation (with over 20 years of continuous task).Fil: Adrados, B.. Universidad Politécnica de Catalunya; EspañaFil: Arias, C. A.. University Aarhus; DinamarcaFil: Perez, Leonardo Martin. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Facultad de Química e Ingeniería-Rosario; Argentina. Universidad Politécnica de Catalunya; España. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Codony, F.. Laboratori Municipal "Aigües de Mataró"; EspañaFil: Bécares, E.. Universidad de León; EspañaFil: Brix, H.. University Aarhus; DinamarcaFil: Morató Farreras, Jordi. Universidad Politécnica de Catalunya; Españ

Genetic and pharmacological inhibition of XBP1 protects against APAP hepatotoxicity through the activation of autophagy

Acetaminophen (APAP) hepatotoxicity induces endoplasmic reticulum (ER) stress which triggers the unfolded protein response (UPR) in hepatocytes. However, the mechanisms underlying ER stress remain poorly understood, thus reducing the options for exploring new pharmacological therapies for patients with hyperacute liver injury. Eight-to-twelve-week-old C57BL/6J Xbp1-floxed (Xbp1f/f) and hepatocyte-specific knockout Xbp1 mice (Xbp1?hepa) were challenged with either high dose APAP [500?mg/kg] and sacrificed at early (1-2?h) and late (24?h) stages of hepatotoxicity. Histopathological examination of livers, immunofluorescence and immunohistochemistry, Western blot, real time (RT)-qPCR studies and transmission electron microscopy (TEM) were performed. Pharmacological inhibition of XBP1 using pre-treatment with STF-083010 [STF, 75?mg/kg] and autophagy induction with Rapamycin [RAPA, 8?mg/kg] or blockade with Chloroquine [CQ, 60?mg/kg] was also undertaken in vivo. Cytoplasmic expression of XBP1 coincided with severity of human and murine hyperacute liver injury. Transcriptional and translational activation of the UPR and sustained activation of JNK1/2 were major events in APAP hepatotoxicity, both in a human hepatocytic cell line and in a preclinical model. Xbp1?hepa livers showed decreased UPR and JNK1/2 activation but enhanced autophagy in response to high dose APAP. Additionally, blockade of XBP1 splicing by STF, mitigated APAP-induced liver injury and without non-specific off-target effects (e.g., CYP2E1 activity). Furthermore, enhanced autophagy might be responsible for modulating CYP2E1 activity in Xbp1?hepa animals. Genetic and pharmacological inhibition of Xbp1 specifically in hepatocytes ameliorated APAP-induced liver injury by enhancing autophagy and decreasing CYP2E1 expression. These findings provide the basis for the therapeutic restoration of ER stress and/or induction of autophagy in patients with hyperacute liver injury.© 2022. The Author(s)

Novel approach identifies SNPs in SLC2A10 and KCNK9 with evidence for parent-of-origin effect on body mass index.

The phenotypic effect of some single nucleotide polymorphisms (SNPs) depends on their parental origin. We present a novel approach to detect parent-of-origin effects (POEs) in genome-wide genotype data of unrelated individuals. The method exploits increased phenotypic variance in the heterozygous genotype group relative to the homozygous groups. We applied the method to >56,000 unrelated individuals to search for POEs influencing body mass index (BMI). Six lead SNPs were carried forward for replication in five family-based studies (of ∼4,000 trios). Two SNPs replicated: the paternal rs2471083-C allele (located near the imprinted KCNK9 gene) and the paternal rs3091869-T allele (located near the SLC2A10 gene) increased BMI equally (beta = 0.11 (SD), P<0.0027) compared to the respective maternal alleles. Real-time PCR experiments of lymphoblastoid cell lines from the CEPH families showed that expression of both genes was dependent on parental origin of the SNPs alleles (P<0.01). Our scheme opens new opportunities to exploit GWAS data of unrelated individuals to identify POEs and demonstrates that they play an important role in adult obesity

The parent-child relationship and adolescent alcohol use: a systematic review of longitudinal studies

Background: Alcohol use among adolescents has become a major public health problem in the past decade and has large short- and long-term consequences on their health. The aim of this systematic review was to provide an overview of longitudinal cohort studies that have analyzed the association between the parent-child relationship (PCR) and change in alcohol use during adolescence. Methods: A search of the literature from 1985 to July 2011 was conducted in Medline, PsycINFO, and EMBASE in order to identify longitudinal, general population studies regarding the influence of the PCR on alcohol use during adolescence. The studies were screened, and the quality of the relevant studies was assessed. A best-evidence synthesis was used to summarize the results. Results: Twenty-eight relevant studies were identified. Five studies found that a negative PCR was associated with higher levels of alcohol use. Another seven papers only found this association for certain subgroups such as boys or girls, or a specific age group. The remaining sixteen studies did not find any association. Conclusions: We found weak evidence for a prospective association between the PCR and adolescent alcohol use. Further research to the association of the PCR with several types of alcohol use (e.g., initiation or abuse) and to the potential reversed causality of the PCR and alcohol use is required