Suitability Of Nitisinone In Alkaptonuria 1 (SONIA 1): an international, multicentre, randomised, open-label, no-treatment controlled, parallel-group, dose-response study to investigate the effect of once daily nitisinone on 24-h urinary homogentisic acid excretion in patients with alkaptonuria after 4 weeks of treatment.

BACKGROUND: Alkaptonuria (AKU) is a serious genetic disease characterised by premature spondyloarthropathy. Homogentisate-lowering therapy is being investigated for AKU. Nitisinone decreases homogentisic acid (HGA) in AKU but the dose-response relationship has not been previously studied. METHODS: Suitability Of Nitisinone In Alkaptonuria 1 (SONIA 1) was an international, multicentre, randomised, open-label, no-treatment controlled, parallel-group, dose-response study. The primary objective was to investigate the effect of different doses of nitisinone once daily on 24-h urinary HGA excretion (u-HGA24) in patients with AKU after 4 weeks of treatment. Forty patients were randomised into five groups of eight patients each, with groups receiving no treatment or 1 mg, 2 mg, 4 mg and 8 mg of nitisinone. FINDINGS: A clear dose-response relationship was observed between nitisinone and the urinary excretion of HGA. At 4 weeks, the adjusted geometric mean u-HGA24 was 31.53 mmol, 3.26 mmol, 1.44 mmol, 0.57 mmol and 0.15 mmol for the no treatment or 1 mg, 2 mg, 4 mg and 8 mg doses, respectively. For the most efficacious dose, 8 mg daily, this corresponds to a mean reduction of u-HGA24 of 98.8% compared with baseline. An increase in tyrosine levels was seen at all doses but the dose-response relationship was less clear than the effect on HGA. Despite tyrosinaemia, there were no safety concerns and no serious adverse events were reported over the 4 weeks of nitisinone therapy. CONCLUSIONS: In this study in patients with AKU, nitisinone therapy decreased urinary HGA excretion to low levels in a dose-dependent manner and was well tolerated within the studied dose range. TRIAL REGISTRATION NUMBER: EudraCT number: 2012-005340-24. Registered at ClinicalTrials.gov: NCTO1828463

Homogentisic acid is not only eliminated by glomerular filtration and tubular secretion but also produced in the kidney in alkaptonuria.

The clinical effects of alkaptonuria (AKU) are delayed and ageing influences disease progression. Morbidity of AKU is secondary to high circulating homogentisic acid (HGA) and ochronosis. It is not known whether HGA is produced by or processed in the kidney in AKU. Data from AKU patients from four studies were merged to form a single AKU group. A control group of non-AKU subjects was generated by merging data from two non-AKU studies. Data were used to derive renal clearance and fractional excretion (FE) ratios for creatinine, HGA, phenylalanine (PHE) and tyrosine (TYR) using standard calculations, for comparison between the AKU and the control groups. There were 225 AKU patients in the AKU group and 52 in the non-AKU control group. Circulating HGA increased with age (P < 0.001), and was significantly associated with decreased HGA clearance (CLHGA ) (P < 0.001) and FEHGA (P < 0.001). CLHGA and FEHGA were increased beyond the theoretical maximum renal plasma flow, confirming renal production and emphasising the greater contribution of net tubular secretion than glomerular filtration to renal elimination of HGA. The kidneys are crucial to elimination of HGA. Elimination of HGA is impaired with age resulting in worsening disease over time. The kidney is an important site for production of HGA. Tubular secretion of HGA contributes more to elimination of HGA in AKU than glomerular filtration

Loss of imprinting of IGF2 correlates with hypermethylation of the H19 differentially methylated region in hepatoblastoma

IGF2, a maternally imprinted foetal growth factor gene, is implicated in many childhood tumours including hepatoblastoma (HB); however, the genetic and epigenetic alterations have not comprehensively been studied. We analysed the methylation status of the H19 differentially methylated region (DMR), loss of heterozygosity (LOH) and allelic expression of IGF2 in 54 HB tumours, and found that 12 tumours (22%) with LOH, 9 (17%) with loss of imprinting (LOI) and 33 (61%) with retention of imprinting (ROI). Biallelic and monoallelic IGF2 expressions correlated with hypermethylation and normal methylation of H19 DMR, respectively, in two tumours with LOI and seven tumours with ROI. Quantitative RT–PCR analysis showed minimal expression of H19 mRNA and substantial expression of IGF2 mRNA in tumours with LOH or LOI, and substantial expression of both H19 and IGF2 mRNAs in tumours with ROI. Increased IGF2 expression with predominant embryonic P3 transcript was found in the majority of HBs with ROI and foetal livers. In contrast to the earlier reports, our findings suggest that the disruption of the enhancer competition model reported in Wilms' tumour may also occur in HB. Both frequencies of LOH and LOI seem to be lower in HB than in Wilms' tumour, reflecting the different tissue origins

Identification of gene expression changes associated with the initiation of diapause in the brain of the cotton bollworm, Helicoverpa armigera

<p>Abstract</p> <p>Background</p> <p>Diapause, a state of arrested development accompanied by a marked decrease of metabolic rate, helps insects to overcome unfavorable seasons. <it>Helicoverpa armigera </it>(Har) undergoes pupal diapause, but the molecular mechanism of diapause initiation is unclear. Using suppression subtractive hybridization (SSH), we investigated differentially expressed genes in diapause- and nondiapause-destined pupal brains at diapause initiation.</p> <p>Results</p> <p>We constructed two SSH libraries (forward, F and reverse, R) to isolate genes that are up-regulated or down-regulated at diapause initiation. We obtained 194 unique sequences in the F library and 115 unique sequences in the R library. Further, genes expression at the mRNA and protein level in diapause- and nondiapause-destined pupal brains were confirmed by RT-PCR, Northern blot or Western blot analysis. Finally, we classified the genes and predicted their possible roles at diapause initiation.</p> <p>Conclusion</p> <p>Differentially expressed genes at pupal diapause initiation are possibly involved in the regulation of metabolism, energy, stress resistance, signaling pathways, cell cycle, transcription and translation.</p

Unequal allelic expression of wild-type and mutated β-myosin in familial hypertrophic cardiomyopathy

Familial hypertrophic cardiomyopathy (FHC) is an autosomal dominant disease, which in about 30% of the patients is caused by missense mutations in one allele of the β-myosin heavy chain (β-MHC) gene (MYH7). To address potential molecular mechanisms underlying the family-specific prognosis, we determined the relative expression of mutant versus wild-type MYH7-mRNA. We found a hitherto unknown mutation-dependent unequal expression of mutant to wild-type MYH7-mRNA, which is paralleled by similar unequal expression of β-MHC at the protein level. Relative abundance of mutated versus wild-type MYH7-mRNA was determined by a specific restriction digest approach and by real-time PCR (RT-qPCR). Fourteen samples from M. soleus and myocardium of 12 genotyped and clinically well-characterized FHC patients were analyzed. The fraction of mutated MYH7-mRNA in five patients with mutation R723G averaged to 66 and 68% of total MYH7-mRNA in soleus and myocardium, respectively. For mutations I736T, R719W and V606M, fractions of mutated MYH7-mRNA in M. soleus were 39, 57 and 29%, respectively. For all mutations, unequal abundance was similar at the protein level. Importantly, fractions of mutated transcripts were comparable among siblings, in younger relatives and unrelated carriers of the same mutation. Hence, the extent of unequal expression of mutated versus wild-type transcript and protein is characteristic for each mutation, implying cis-acting regulatory mechanisms. Bioinformatics suggest mRNA stability or splicing effectors to be affected by certain mutations. Intriguingly, we observed a correlation between disease expression and fraction of mutated mRNA and protein. This strongly suggests that mutation-specific allelic imbalance represents a new pathogenic factor for FHC

Neurite Outgrowth Stimulated by Oxytocin Is Modulatedby Inhibition of the Calcium Voltage-Gated Channels

Abstract Neuropeptide oxytocin contributes to the regulationof the neuron differentiation and cell morphology.However, the precise mechanisms are not yet fullyunderstood. Oxytocin receptor function and its coupling tocalcium entry are obvious objects of interest in relation tothe neuron morphology. Postsynaptic scaffolding proteinsincluding SHANK proteins interact with other synapticmolecules and change dendritic morphology. SH-SY5Yneuroblastoma cell line represents a useful neurobiologicalin vitro model to study the short-term oxytocin effects onneurite outgrowth and underlying mechanisms. In thepresent study, we show that oxytocin induces an increase inthe intracellular calcium in SH-SY5Y cells. Specificity ofthe calcium influx was verified by blockade of the oxytocinreceptors with oxytocin receptor antagonist L-371,257.Neurite outgrowth stimulated by oxytocin was inhibited byspecific voltage-gated calcium channel blockers. Theexposure of SH-SY5Y cells to oxytocin resulted in a significantincrease in the gene expression of SHANK1 andSHANK3 proteins. Overall, the present data indicate thatoxytocin may contribute to the regulation of scaffoldingproteins expression known to be associated with clusters ofcalcium channels at the cell membrane. It appears thatoxytocin stimulated neurite outgrowth is, at least, in partdependent on the voltage-gated calcium channels.Keywords Oxytocin receptor Neurite outgrowth SHANK Development Calciu

Comparison of self-stigma and quality of life in patients with depressive disorders and schizophrenia spectrum disorders &ndash; a cross-sectional study

Michaela Holubova,1,2 Jan Prasko,1 Stanislav Matousek,1 Klara Latalova,1 Marketa Marackova,1 Kristyna Vrbova,1 Ale&scaron; Grambal,1 Milos Slepecky,3 Marta Zatkova3 1Department of Psychiatry, Faculty of Medicine and Dentistry, Palacky University Olomouc, University Hospital Olomouc, Olomouc, 2Department of Psychiatry, Hospital Liberec, Liberec, Czech Republic; 3Department of Psychology Sciences, Faculty of Social Science and Health Care, Constantine the Philosopher University in Nitra, Nitra, Slovak Republic Background: The views of one&rsquo;s self-stigma and quality of life (QoL) in patients with schizophrenia and depressive disorders are significant subjective notions, both being proven to affect patient&rsquo;s functioning in life. The objective of this study was to investigate the QoL and self-stigma in connection with demographic factors and compare the two groups of patients in terms of those variables. Methods: In a cross-sectional study, the outpatients with schizophrenia spectrum disorders and depressive disorders completed the Quality of Life Satisfaction and Enjoyment Questionnaire, the Internalized Stigma of Mental Illness Scale, and a demographic questionnaire during a routine psychiatric control. Furthermore, both patients and their psychiatrists evaluated the severity of the disorder by Clinical Global Impression-Severity scale. Results: The QoL of patients with depressive disorders or schizophrenia spectrum disorders did not significantly differ between the two groups. In both groups, unemployment was perceived to be a significant factor decreasing the QoL. Self-stigma was detected to be higher in patients with schizophrenia spectrum disorders than in patients with depressive disorders. A strong correlation was found between the two scales, meaning that those with higher levels of self-stigmatization were less prone to see their life as fulfilling and joyful. Conclusion: This study shows that the degree of the internalized stigma can be an important aspect linked to the QoL irrespective of the diagnostic category. Keywords: self-stigma, quality of life, depressive disorders, schizophrenia spectrum disorder

Coping, schemas, and cardiovascular risks &ndash; study protocol

Milos Slepecky,1 Antonia Kotianova,1 Jan Prasko,2 Ivan Majercak,3 Erika Gyorgyova,4 Michal Kotian,1,5 Marta Zatkova,1 Ingrid Tonhajzerova,6,7 Michaela Chupacova,1,5 Marta Popelkova1 1Department of Psychology Sciences, Faculty of Social Science and Health Care, Constantine the Philosopher University in Nitra, Nitra, Slovak Republic; 2Department of Psychiatry, Faculty of Medicine and Dentistry, University Palacky Olomouc, University Hospital Olomouc, Olomouc, Czech Republic; 31st Department of Internal Medicine, Faculty of Medicine, Pavol Josef Safarik University in Ko&scaron;ice, 4Internal Medicine and Cardiology Private Practice MUDr Ivan Majercak, Ko&scaron;ice, 5Psychagogia, Liptovsky Mikulas, 6Department of Physiology, 7Biomedical Center Martin, Jessenius Faculty of Medicine in Martin, Comenius University in Bratislava, Martin, Slovak Republic Abstract: The aim of this article is to describe the protocol of a trial focusing on the psychological, anthropometric, cardiac, and psychophysiological factors contributing to increased risk of cardiovascular diseases (CVDs). As background, the article provides a short overview of research literature linking personal traits, maladaptive schemas, and coping styles with CVDs through reactivity of the autonomic nervous system. Keywords: early maladaptive schemas, coping styles, personality traits, dissociation, heart rate variabilit