Functional Inactivation of Drosophila GCK Orthologs Causes Genomic Instability and Oxidative Stress in a Fly Model of MODY-2.

Maturity-onset diabetes of the young (MODY) type 2 is caused by heterozygous inactivating mutations in the gene encoding glucokinase (GCK), a pivotal enzyme for glucose homeostasis. In the pancreas GCK regulates insulin secretion, while in the liver it promotes glucose utilization and storage. We showed that silencing the Drosophila GCK orthologs Hex-A and Hex-C results in a MODY-2-like hyperglycemia. Targeted knock-down revealed that Hex-A is expressed in insulin producing cells (IPCs) whereas Hex-C is specifically expressed in the fat body. We showed that Hex-A is essential for insulin secretion and it is required for Hex-C expression. Reduced levels of either Hex-A or Hex-C resulted in chromosome aberrations (CABs), together with an increased production of advanced glycation end-products (AGEs) and reactive oxygen species (ROS). This result suggests that CABs, in GCK depleted cells, are likely due to hyperglycemia, which produces oxidative stress through AGE metabolism. In agreement with this hypothesis, treating GCK-depleted larvae with the antioxidant vitamin B6 rescued CABs, whereas the treatment with a B6 inhibitor enhanced genomic instability. Although MODY-2 rarely produces complications, our data revealed the possibility that MODY-2 impacts genome integrity.

Pyridoxine/pyridoxamine 5'-phosphate oxidase (Sgll/PNPO) is important for DNA integrity and glucose homeostasis maintenance in Drosophila.

Pyridoxine/pyridoxamine 5'-phosphate oxidase (PNPO) and pyridoxal kinase (PDXK) cooperate to produce pyridoxal 5'-phosphate (PLP), the active form of vitamin B6. PDXK phosphorylates pyridoxine, pyridoxamine, and pyridoxal by producing PNP, PMP, and PLP, whereas PNPO oxidizes PNP, PMP, into PLP. We previously demonstrated that PDXK depletion in Drosophila and human cells impacts on glucose metabolism and DNA integrity. Here we characterized sgll, the Drosophila ortholog of PNPO gene, showing that its silencing by RNA interference elicits chromosome aberrations (CABs) in brains and induces diabetic hallmarks such as hyperglycemia and small body size. We showed that in sgllRNAi neuroblasts CABs are largely produced by the genotoxic effect of the advanced glycation end products triggered by high glucose. As in sgllRNAi cells, part of PLP is still produced by PDXK activity, these data suggest that PLP dosage need to be tightly regulated to guarantee glucose homeostasis and DNA integrity.

The Association among Default Mode Network Functional Connectivity, Mentalization, and Psychopathology in a Nonclinical Sample: An eLORETA Study.

AIMS: We investigated default mode network (DMN) electroencephalography (EEG) functional connectivity differences between individuals with self-reported high mentalization capability and low psychopathological symptoms, versus participants with mentalization impairments and high psychopathological symptoms. METHODS: Forty-nine students (35 women) with a mean age of 22.92 ± 2.53 years were administered the Mentalization Questionnaire (MZQ) and the Symptom Checklist-90-Revised. Five minutes of EEG during resting state were also recorded for each participant. DMN functional connectivity analyses were conducted by means of the exact Low Resolution Electric Tomography software (eLORETA). RESULTS: Compared to the individuals with high mentalization capability and lower self-reported psychopathological symptoms, participants with mentalization impairments and high psychopathological symptoms showed a decrease of EEG beta connectivity between: (i) the right and left medial frontal lobe, and (ii) the left medial frontal lobe and the right anterior cingulate cortex. Furthermore, while MZQ total score was positively associated with DMN network connections (i.e., right and left medial frontal lobes), several psychopathological symptoms (i.e., interpersonal sensitivity, depression, and psychoticism) were negatively associated with DMN connectivity. CONCLUSION: Our results may reflect a top-down emotion regulation deficit which is associated with both internalizing and externalizing behavior problems.

Alpha/Theta Neurofeedback Increases Mentalization and Default Mode Network Connectivity in a Non-Clinical Sample.

Several studies showed the effectiveness of alpha/theta (A/T) neurofeedback training in treating some psychiatric conditions. Despite the evidence of A/T effectiveness, the psychological and neurobiological bases of its effects is still unclear. The aim of the present study was to explore the usefulness of the A/T training in increasing mentalization in a non-clinical sample. The modifications of electroencephalographic (EEG) functional connectivity in Default Mode Network (DMN) associated with A/T training were also investigated. Forty-four subjects were enrolled in the study and randomly assigned to receive ten sessions of A/T training [neurofeedback group (NFG) = 22], or to act as controls [waiting list group (WLG) = 22]. All participants were administered the mentalization questionnaire (MZQ) and the Symptom Checklist-90-Revised (SCL-90-R). In the post training assessment, compared to WLG, NFG showed a significant increase of MZQ total scores (3.94 ± 0.73 vs. 3.53 ± 0.77; F1;43 = 8.19; p = 0.007; d = 0.863). Furthermore, A/T training was also associated with a significant increase of EEG functional connectivity in several DMN brain areas (e.g. Posterior Cingulate Cortex). Taken together our results support the usefulness of the A/T training in enhancing mentalization and DMN connectivity.

The expression of four pyridoxal kinase (PDXK) human variants in Drosophila impacts on genome integrity.

In eukaryotes, pyridoxal kinase (PDXK) acts in vitamin B6 salvage pathway to produce pyridoxal 5'-phosphate (PLP), the active form of the vitamin, which is implicated in numerous crucial metabolic reactions. In Drosophila, mutations in the dPdxk gene cause chromosome aberrations (CABs) and increase glucose content in larval hemolymph. Both phenotypes are rescued by the expression of the wild type human PDXK counterpart. Here we expressed, in dPdxk1 mutant flies, four PDXK human variants: three (D87H, V128I and H246Q) listed in databases, and one (A243G) found in a genetic screening in patients with diabetes. Differently from human wild type PDXK, none of the variants was able to completely rescue CABs and glucose content elicited by dPdxk1 mutation. Biochemical analysis of D87H, V128I, H246Q and A243G proteins revealed reduced catalytic activity and/or reduced affinity for PLP precursors which justify this behavior. Although these variants are rare in population and carried in heterozygous condition, our findings suggest that in certain metabolic contexts and diseases in which PLP levels are reduced, the presence of these PDXK variants could threaten genome integrity and increase cancer risk.

Coping food craving with neurofeedback. Evaluation of the usefulness of alpha/theta training in a non-clinical sample.

The aim of the present study was to explore the usefulness of the alpha/theta (A/T) training in reducing Food Craving (FC) in a non-clinical sample. The modifications of electroencephalographic (EEG) power spectra associated with A/T training was also investigated. Fifty subjects were enrolled in the study and randomly assigned to receive ten sessions of A/T training [neurofeedback group (NFG)=25], or to act as controls [waiting list group (WLG)=25]. All participants were administered the Food Cravings Questionnaire-Trait, the Eating Disorder Examination Questionnaire and the Symptom Checklist-90-Revised. In the post training assessment, compared to the WLG, the NFG showed a significant reduction of intentions and plans to consume food (F1; 49=4.90; p=.033; d=0.626) and of craving as a physiological state (F1; 49=8.09; p=.007; d=803). In NFG, changes in FC persisted after 4months follow-up. Furthermore, A/T training was associated with significant a increase of resting EEG alpha power in several brain areas involved in FC (e.g., insula) and food cue reactivity (e.g., parahippocampal gyrus, inferior and superior temporal gyrus). Taken together, our results showed that ten sessions of A/T training are associated with a decrease of self-reported FC in a non-clinical sample. These findings suggest that this brain-directed intervention may be useful in the treatment of dysfunctional eating behaviors characterized by FC.

Default Mode Network alterations in alexithymia: an EEG power spectra and connectivity study.

Recent neuroimaging studies have shown that alexithymia is characterized by functional alterations in different brain areas [e.g., posterior cingulate cortex (PCC)], during emotional/social tasks. However, only few data are available about alexithymic cortical networking features during resting state (RS). We have investigated the modifications of electroencephalographic (EEG) power spectra and EEG functional connectivity in the default mode network (DMN) in subjects with alexithymia. Eighteen subjects with alexithymia and eighteen subjects without alexithymia matched for age and gender were enrolled. EEG was recorded during 5 min of RS. EEG analyses were conducted by means of the exact Low Resolution Electric Tomography software (eLORETA). Compared to controls, alexithymic subjects showed a decrease of alpha power in the right PCC. In the connectivity analysis, compared to controls, alexithymic subjects showed a decrease of alpha connectivity between: (i) right anterior cingulate cortex and right PCC, (ii) right frontal lobe and right PCC, and (iii) right parietal lobe and right temporal lobe. Finally, mediation models showed that the association between alexithymia and EEG connectivity values was directed and was not mediated by psychopathology severity. Taken together, our results could reflect the neurophysiological substrate of some core features of alexithymia, such as the impairment in emotional awareness.