Uridine and pyruvate protect T cells' proliferative capacity from mitochondrial toxic antibiotics: a clinical pilot study.

Antibiotics that inhibit bacterial protein or nucleic acid synthesis and function can exert an off-target action on mitochondria (mitotoxic antibiotics), making actively dividing mammalian cells dependent on uridine and pyruvate supplementation. Based on this rationale, we carried out, for the first time, a randomized pilot study in 55 patients with asymptomatic bacteriuria or positive sperm culture, each treated with a single mitotoxic antibiotic with or without oral supplementation of uridine + pyruvate (Uripyr, Mitobiotix, Italy). The in vivo and ex vivo data show a a 3.4-fold higher value in the differential (before and after the antibiotic treatment) lymphocytes count and a 3.7-fold increase in the percentage of dividing T cells, respectively, in the Uripyr vs the control group. Our findings lay the groundwork to enhance the synergy between antibiotics and the immune system in order to optimize the administration protocols and widen the application potentials of antibiotic therapies as well as to re-evaluate old "forgotten" molecules to fight bacterial infections in the antibiotics resistance era.

A systematic review of person-centered approaches to investigating patterns of trauma exposure.

Recent research has found that exposure to traumatic events may occur in certain patterns, rather than randomly. Person-centered analyses, and specifically latent class analysis, is becoming increasingly popular in examining patterns, or 'classes' of trauma exposure. This review aimed to identify whether there are consistent homogeneous subgroups of trauma-exposed individuals, and the relationship between these trauma classes and psychiatric diagnosis. A systematic review of the literature was completed using the databases EMBASE, MEDLINE (PubMed) and PsycINFO. From an initial yield of 189, 17 studies met inclusion criteria. All studies identified a group of individuals who had a higher likelihood of exposure to a wide range of traumas types, and this group consistently exhibited worse psychiatric outcomes than other groups. Studies differed in the nature of the other groups identified although there was often a class with high levels of sexual interpersonal trauma exposure, and a class with high levels of non-sexual interpersonal trauma. There was some evidence that risk for psychiatric disorder differed across these classes. Person-centered approaches to understanding the relationship between trauma exposure and mental health may offer ways to improve our understanding of the role trauma exposure plays in increasing vulnerability to psychiatric disorder.

Assessment of ABCG2-mediated transport of xenobiotics across the blood-milk barrier of dairy animals using a new MDCKII in vitro model.

The ATP-binding cassette (ABC) efflux transporter ABCG2 represents the main route for active secretion of drugs and toxins across the blood-milk barrier, thereby producing a potential health risk for dairy consumers through formation of relevant residues in milk. However, no suitable in vitro model is as yet available to systematically investigate ABCG2-mediated transport of xenobiotics into milk of dairy animals. We recently cloned ABCG2 from the lactating mammary gland of dairy cows (bABCG2) and goats (cABCG2). Thus, the objective of this study was to generate a suitable blood-milk barrier in vitro model using polarized MDCKII monolayers stably expressing mammary bABCG2 or cABCG2. ABCG2 protein was localized by confocal microscopy to the apical and lateral plasma membrane of polarized MDCKII cells. Intact barrier function of MDCKII-bABCG2 and MDCKII-cABCG2 monolayers was confirmed by determination of cell permeability of transcellular marker propranolol and paracellular marker atenolol which was ≤1 %. In flux assays, ABCG2 substrate 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) showed preferential basolateral to apical (B > A) transport in ABCG2-MDCKII cells. This apically directed PhIP transport was significantly inhibited by ABCG2 inhibitor fumitremorgin C (FTC) or the flavonoid equol. PhIP B > A transport in MDCKII-bABCG2 monolayers was additionally decreased by ABCG2 inhibitor Ko143. The fluoroquinolone antibiotic enrofloxacin was identified as a substrate of ruminant mammary ABCG2. The analgesic drug sodium salicylate was shown to be substrate of bABCG2 but not of cABCG2. Thus, the generated mammary ABCG2-expressing MDCKII cells represent a valuable tool to study active secretion of drugs and toxins into milk.

Antiepileptic drugs reduce the efficacy of methotrexate chemotherapy through accelerated degradation of the reduced folate carrier by the ubiquitin-proteasome pathway.

BACKGROUND/AIMS: Concurrent treatment with methotrexate (MTX) and enzyme-inducing antiepileptic drugs including phenobarbital (PB) reduces the efficacy of MTX chemotherapy in cancer patients. We have shown that Reduced folate carrier (Rfc1)-mediated uptake of MTX, an essential determinant of MTX chemotherapy, is significantly reduced by PB via protein kinase C (PKC). However, whether PB treatment affects Rfc1 activity through regulation of carrier protein stability and the mechanisms involved remain unclear. METHODS/RESULTS: Protein turnover assays using hepatocytoma cells demonstrated that Rfc1 is a long-lived protein that is mainly degraded by the ubiquitin-proteasome proteolytic pathway under basal conditions. Pretreatment with PB significantly reduced Rfc1-mediated MTX uptake and shortened the carrier protein half-life. This effect was abolished by the specific PKC inhibitor Gö6976. Inhibition of proteasomes with MG-132 significantly elevated Rfc1 protein levels and induced colocalization of Rfc1 and ubiquitin particularly in submembranous cellular compartments. Finally, we demonstrated that PB treatment resulted in enhanced levels of Rfc1 polyubiquitin conjugates. CONCLUSIONS: Our results demonstrate that PB treatment causes downregulation of Rfc1 activity through PKC-dependent accelerated degradation of the Rfc1 protein by the ubiqutin-proteasome pathway. This regulatory mechanism may therefore involve clinically relevant drug resistance in patients concurrently receiving MTX and enzyme-inducing antiepileptic drugs.

Emergencies related to cocaine use: a European multicentre study of expert interviews.

Illicit drug use can lead to acute reverse reactions leading to the admission to emergency departments. Cocaine-related emergencies have been monitored in the USA, but not in Europe so far. The study investigates patterns of cocaine emergencies in eight European cities in a multicentre cross-sectional study conducted in Barcelona, Budapest, Dublin, Hamburg, London, Rome, Vienna and Zurich. The reported frequency differs from city to city, with some emergency centres having less than one case per half year, and some centres having more than one case per month. Patterns of complaints among cocaine users are associated with the psychomotor-stimulant or cardiovascular effects of cocaine. Urine screens and referrals to the addiction services are infrequent. A closer link between the emergency departments and addiction services would help in guiding problematic drug users towards appropriate treatment at an earlier stage in the addiction process.

Trauma and PTSD in patients with alcohol, drug, or dual dependence: a multi-center study.

BACKGROUND: We investigated (1) the prevalence of posttraumatic stress disorder (PTSD) in treatment-seeking subjects with substance use dependence (SUD), (2) the association between comorbid PTSD and the severity and course of addiction and psychopathology, and (3) this association in patients with subsyndromal PTSD, and in trauma exposure without PTSD. METHODS: In this cross-sectional study, 459 subjects in 14 German addiction treatment centers participated with alcohol-dependence (A) in 39.7%, drug-dependence (D) in 33.6%, or both (AD) 26.8%. The diagnostic measures included the International Diagnostic Checklists (IDCL), Posttraumatic Diagnostic Scale (PDS), Addiction Severity Index (ASI), and the Brief Psychiatric Rating Scale (BPRS). Associations between independent characteristics and outcomes were analysed by univariate and multivariate statistics. RESULTS: 25.3% of the subjects had PTSD confirmed by both IDCL and PDS with higher rates in the AD (34.1%) and D (29.9%) groups compared with group A (15.4%, p < 0.001). In 22.8%, PTSD was subsyndromal (either IDCL or PDS positive) without significant differences between SUD groups, and 18.3% met PTSD trauma criteria A without PTSD (exposure). After controlling for SUD and gender, trauma subgroups significantly differed regarding the onset of alcohol-related symptoms (p < 0.02), numbers of previous admissions (p < 0.03), severity of SUD (p < 0.001), current craving (p < 0.02), and psychopathology (p < 0.001). We observed the worst outcome in PTSD, while trauma exposure had no effects. CONCLUSIONS: The prevalence of PTSD is higher in drug than in alcohol dependence. The more strictly PTSD is diagnosed (by interviewer and questionnaire) the more clearly are associations with characteristics of SUD. PTSD seems to be an independent risk factor for an unfavorable outcome of SUD.