Cadmium chloride inhibits lactate gluconeogenesis in isolated human renal proximal tubules: a cellular metabolomic approach with <Superscript>13</Superscript>C-NMR

As part of a study on cadmium nephrotoxicity, we studied the effect of cadmium chloride (CdCl₂) in isolated human renal proximal tubules metabolizing the physiological substrate lactate. Dose–effect experiments showed that 10–500 μM CdCl₂ reduced lactate removal, glucose production and the cellular levels of ATP, coenzyme A, acetyl-coenzyme A and of reduced glutathione in a dose-dependent manner. After incubation with 5 mM l-[1-¹³C]-, or l-[2-¹³C]-, or l-[3-¹³C] lactate or 5 mM l-lactate plus 25 mM NaH¹³CO₃ as substrates, substrate utilization and product formation were measured by both enzymatic and carbon 13 NMR methods. Combination of enzymatic and NMR measurements with a mathematical model of lactate metabolism previously validated showed that 100 μM CdCl₂ caused an inhibition of flux through lactate dehydrogenase and alanine aminotransferase and through the entire gluconeogenic pathway; fluxes were diminished by 19% (lactate dehydrogenase), 28% (alanine aminotransferase), 28% (pyruvate carboxylase), 42% (phosphoenolpyruvate carboxykinase), and 52% (glucose-6-phosphatase). Such effects occurred without altering the oxidation of the lactate carbons or fluxes through enzymes of the tricarboxylic acid cycle despite a large fall of the cellular ATP level, a marker of the energy status and of the viability of the renal cells. These results that were observed at clinically relevant tissue concentrations of cadmium provide a biochemical basis for a better understanding of the cellular mechanism of cadmium-induced renal proximal tubulopathy in humans chronically exposed to cadmium.     

Stability-indicating micellar electrokinetic chromatography method for the analysis of sumatriptan succinate in pharmaceutical formulations

A micellar electrokinetic chromatography method for the determination of sumatriptan succinate in pharmaceutical formulations was developed. The effects of several factors such as pH, surfactant and buffer concentration, applied voltage, capillary temperature, and injection time were investigated. Separation took about 5 min using phenobarbital as internal standard. The separation was carried out in reversed polarity mode at 20 °C, 26 kV and using hydrodynamic injection for 10 s. Separation was achieved using a bare fused-silica capillary 50 μm × 40 cm and background electrolyte of 25 mM sodium dihydrogen phosphate—adjusted with concentrated phosphoric acid to pH 2.2, containing 125 mM sodium dodecyl sulfate and detection was at 226 nm. The method was validated with respect to linearity, limits of detection and quantification, accuracy, precision and selectivity. The calibration curve was linear over the range of 100-2000 μg mL⁻¹. The relative standard deviations of intra-day and inter-day precision for migration time, peak area, corrected peak area, ratio of corrected peak area and ratio of peak area were less than 0.68, 3.48, 3.28, 2.97 and 2.83% and 2.01, 5.50, 4.46, 4.92 and 4.07%, respectively. The proposed method was successfully applied to the determinations of the analyte in tablet. Forced degradation studies were conducted by introducing a sample of sumatriptan succinate standard solution to different forced degradation conditions using neutral (water), basic (0.1 M NaOH), acidic (0.1 M HCl), oxidative (10% H₂O₂) and photolytic (exposure to UV light at 254 nm for 2 h). It is concluded that the stability-indicating method for sumatriptan succinate can be used for the analysis of the drug in various samples.     

Protective potential of MMR vaccine against complete Freund’s adjuvant-induced inflammation in rats

The aim of the present study was to investigate the effect of MMR vaccine on inflammation which was induced by complete Freund’s adjuvant (CFA) in male Sprague–Dawley rats. Rats were randomly divided into the control, CFA, MMR and CFA + MMR groups. Inflammatory symptoms such as paw oedema was measured in CFA-injected rats’ paw. Body weight changes and alterations in some haematological parameters and oxidative stress markers following CFA injection were checked. In CFA-inflammed rats, there was a significant increase in rat paw thickness and decrease in body weight increment. MMR exhibited a significant anti-inflammatory effect as manifested by reduction in paw thickness and normal gain in body weight when administered 1 week prior to induction of inflammation. The altered haematological parameters (TLC) and oxidative stress markers (MDA, GSH, SOD) in the inflammed rats were significantly brought back to near normal by MMR treatment. In conclusion, MMR vaccine showed a reduction in rat paw thickness and it could significantly normalize the haematological and biochemical abnormalities in CFA-induced inflammatory pain model in rats. Our data suggested that MMR could be a potential protective agent against certain types of inflammatory pain. Further histopathological and radiological studies are required to confirm the possibility of developing novel therapeutic vaccines against some forms of arthritis.     

Immunomodulatory studies of a bioactive fraction from the fruit of Prunus cerasus in BALB/c mice

In order to evaluate the role of ethyl acetate fraction (PNRS-EtOAC) obtained from the Prunus cerasus fruit in the modulation of immune responses, detailed studies were carried out using a panel of in vivo assays. Oral administration of PNRS-EtOAC (25-100 mg/kg) stimulated the IgM and IgG titre expressed in the form of hemagglutination antibody (HA) titre. Further, it elicited a dose related increase in the delayed type hypersensitivity reaction (DTH) after 24 and 48 h in BALB/c mice. Besides augmenting the humoral and cell mediated immune response, the concentration of cytokines (IFN-γ, IL-4, and TNF-α) in serum with respect to T cell interactions, i.e. the proliferation of lymphocytes were significantly increased at 50 mg/kg compared with the control. The results in these studies demonstrated the immunostimulatory effect of PNRS-EtOAC in a dose-dependent manner with respect to the macrophage activation possibly expressing the phagocytosis and nitrite production by the enhancement of TNF-α production as a mode of action.     

The Methylazoxymethanol Acetate (MAM-E17) Rat Model: Molecular and Functional Effects in the Hippocampus

Administration of the DNA-alkylating agent methylazoxymethanol acetate (MAM) on embryonic day 17 (E17) produces behavioral and anatomical brain abnormalities, which model some aspects of schizophrenia. This has lead to the premise that MAM rats are a neurodevelopmental model for schizophrenia. However, the underlying molecular pathways affected in this model have not been elucidated. In this study, we investigated the molecular phenotype of adult MAM rats by focusing on the frontal cortex and hippocampal areas, as these are known to be affected in schizophrenia. Proteomic and metabonomic analyses showed that the MAM treatment on E17 resulted primarily in deficits in hippocampal glutamatergic neurotransmission, as seen in some schizophrenia patients. Most importantly, these results were consistent with our finding of functional deficits in glutamatergic neurotransmission, as identified using electrophysiological recordings. Thus, this study provides the first molecular evidence, combined with functional validation, that the MAM-E17 rat model reproduces hippocampal deficits relevant to the pathology of schizophrenia.     

Early childhood experiences, parenting and the process of drug dependency among young people in Tehran, Iran

Introduction and Aims. Substance abuse has become a major public health problem in Iran. The process of developing an addiction is complex and multifaceted. Early childhood experiences are thought to be one of the important determinants of addictive behaviour. The aim of this qualitative study is to explore the early childhood experiences, especially the experiences within the immediate family, of current substance‐using young adults in Iran. Design and Methods. The study is qualitative in nature. In‐depth interviews were conducted with 15 young men and women who were either in treatment for their addiction or were active drug users at the time of the interviews. Moreover, four interviews have been conducted with family members of participants. Results. The majority of the participants experienced traumatic events during childhood and came from dysfunctional families. There appears to be a significant disconnect between these individuals and their families. An obedience‐instilling parenting style and parents' knowledge and attitude toward drug using and prevention were also identified as important determinants of substance use. Discussion and Conclusions. The results of this research point out the need for early interventions for at‐risk families as well as at‐risk individuals.[Mirlashari J, Demirkol A, Salsali M, Rafiey H, Jahanbani J. Early childhood experiences, parenting and the process of drug dependency among young people in Tehran, Iran. Drug Alcohol Rev 2012;31:461–468]     

Child and adolescent service experience (ChASE): measuring service quality and therapeutic process

OBJECTIVES. Dissatisfaction with services has been associated with poorer child mental health outcomes, early treatment termination as well as disagreements over the nature of mental health difficulties, reasons for referral and therapy goals. The development of straightforward, reliable, and accurate methods of eliciting service users' views is essential within child and adolescent mental health care. This paper describes the development of the child and adolescent service experience (ChASE), a tool to measure children and young people's service experience DESIGN. The study comprises a non-experimental, cross-sectional design. METHODS. Participants were 132 mental health service users aged 8-18 years. Participants and their main carer completed the ChASE, Parent Satisfaction Questionnaire (PSQ) (Stallard, 1996) and Strengths and Difficulties (SDQ) Impact Supplement. Clinicians completed the SDQ Impact Supplement and provided clinical activity data. A sub-sample of participants completed the ChASE on a second occasion, 6 weeks after the completion of the first questionnaire. RESULTS. Scrutiny of ChASE data indicated high levels of completion. Principal axis factoring identified three factors within the ChASE: Relationship, Privacy, and Session Activity. The ChASE has good internal consistency and test-retest reliability. Significant correlations were found between the ChASE and carer satisfaction, service use, and youth clinical outcomes. CONCLUSIONS. The ChASE is a short, psychometrically robust tool for routine measurement of children, and young people's experience of mental health services, which users can complete easily. The results underline the importance of alliance factors to children and young people and their association with clinical improvement as well as the potential for the ChASE to be used a measure of children's therapeutic progress and alliance.     

Hepatitis C infection, Cognition, and inflammation in an Egyptian sample

Chronic hepatitis C (HCV) infection is associated with cognitive impairments which might be mediated through a secondary inflammatory cascade. Egypt has an unusually high prevalence of HCV monoinfections and is an ideal site for the study of the isolated effects of HCV infection. Therefore, in a hospital‐based cross‐sectional study based in Egypt, this study compared cognitive functioning and serum markers of inflammation in 11 HCV positive cases and 14 HCV negative controls. The Wisconsin Card Sorting Test was used to assess cognitive flexibility and the Brief Visuospatial Memory Test‐Revised was used to assess learning and memory. Circulating levels of soluble tumor necrosis factor receptor II (sTNFR‐II), monocyte chemotactic protein‐1 (MCP‐1/CCL2), and soluble CD14 (sCD14) were determined as indices of inflammation. HCV positive cases had higher levels of sTNFR‐II (t = ?3.5, P = 0.002). HCV positive cases also had significantly worse cognitive flexibility with higher number of total errors (t = ?2.18, P = 0.04), and preservative responses (t = ?2.12, P = 0.05), and lower number of conceptual level responses (t = 1.32, P = 0.04) on the Wisconsin Card Sorting test. In conclusion, results from this pilot study indicate that HCV+ patients have worse cognitive performance and somewhat greater inflammatory activity as compared to controls. The increased inflammation may be associated with the cognitive impairments observed in these HCV+ patients. J. Med. Virol. 83:261–266, 2011. © 2010 Wiley‐Liss, Inc.     

Human leukocyte antigen class I polymorphisms influence the mild clinical manifestation of Plasmodium falciparum infection in Ghanaian children

A prospective study that included 429 children for active detection of mild malaria was conducted in a coastal region of Ghana to reveal whether the incidence of malaria is affected by human leukocyte antigen (HLA) polymorphism. During 12 months of follow-up, 85 episodes of mild clinical malaria in 74 individuals were observed, and 34 episodes among them were accompanied with significant parasitemia at >5000 infected red blood cells per cubic millimeter. Attributable and relative risks conferred by genetic factors in the HLA region were evaluated by comparison of the incidence in children, stratified by carrier status, of a given allele of HLA-A, -B, -DRB1 and TNFA promoter polymorphism. HLA-B*35:01 reduced the incidence by 0.178 events per person per year (0.060 versus 0.239 for B*35:01-positive and -negative subpopulations, respectively), and a relative risk of 0.25, which remained statistically significant after Bonferroni's correction for multiple testing (p(c) = 8.2 × 10(-5)). Further, HLA-B*35:01 and -B*53:01 exhibited opposite effects on the incidence of malaria with significant parasitemia. When parasite densities in different HLA carriers status were compared, HLA-A*01 conferred an increase in parasite load (p = 6.0 × 10(-7)). In addition, we found a novel DRB1 allele that appears to have emerged from DRB1*03:02 by single nucleotide substitution.