Ghrelin, peptide YY and their receptors: gene expression in brain from subjects with and without Prader-Willi syndrome

Ghrelin and peptide YY (PYY) are peptides generally produced by the gastrointestinal organs which are involved in appetite regulation via highly specialized centers in the brain. Abnormal plasma ghrelin and PYY levels compared with controls have been reported for subjects with Prader-Willi syndrome (PWS) which is characterized by infantile hypotonia, poor suck reflex and failure to thrive followed by hyperphagia and marked obesity in early childhood. We studied gene expression of ghrelin, peptide YY, and their receptors (i.e., GHS-R1a, GHS-R1b, and NPY2R) in six different brain regions (frontal cortex, temporal cortex, visual cortex, pons, medulla, and hypothalamus) obtained from three subjects with PWS, two individuals with Angelman syndrome, and six controls to determine if expression of these genes is detectable in different regions of the brain in subjects with and without PWS. In general, expression of these genes using RT-PCR was detected in all subjects and no obvious differences were seen in their pattern of expression between subjects with or without PWS. Additional studies including quantitative gene expression measurements will be required to further evaluate the role of these genes in the eating disorder seen in PWS.     

Role of AMPA receptor desensitization and the side effects of a DMSO vehicle on reticulospinal EPSPs and locomotor activity

Activation of the vertebrate locomotor network is mediated by glutamatergic synaptic drive, normally initiated by the brain stem. Previous investigations have studied the role of glutamate receptors, especially NMDA receptors, in generating and regulating locomotor pattern generation. Few studies, however, have focused on the role of AMPA receptors in shaping network activity, especially with regard to their rapid desensitization. It is important to determine whether AMPA receptor desensitization plays a role in regulating neuronal network activity. We examined this question on both the network and synaptic levels in the lamprey (Lampetra fluviatilis) spinal cord using a selective and potent inhibitor of AMPA receptor desensitization, cyclothiazide (CTZ). The solvent dimethyl sulfoxide (DMSO) is commonly used to dissolve this drug, as well as many others. Unexpectedly, the vehicle alone already at 0.02%, but not at 0.01%, caused significant increases in excitatory postsynaptic potential (EPSP) amplitudes and NMDA-induced locomotor frequency. The results indicate that DMSO may have a profound influence when used > or = 0.02%, a concentration 10-50 times less than that most commonly used. Subsequently we applied CTZ concentrations < or =10 microM (DMSO < or =0.01%). CTZ (1.25-5 microM) caused an appreciable and significant increase in EPSPs mediated by non-NMDA receptors and in both AMPA- and NMDA-induced locomotor frequency, but no effects on EPSPs mediated by NMDA receptors. From the effects of CTZ it is apparent that AMPA receptor desensitization plays an important role in determining locomotor frequency and that this is likely a result of its limiting function on AMPA receptor-mediated EPSPs.     

A 9-year-old male with a duplication of chromosome 3p25.3p26.2: clinical report and gene expression analysis

We describe a 9-year-old male referred for genetic evaluation for Prader-Willi syndrome (PWS). PWS is the most common genetically defined cause of life-threatening obesity and results from a functional loss of paternally expressed genes from the chromosome 15q11-q13 region. The patient presented with pervasive developmental disorder, delayed speech, and rapid onset of obesity at age 4 years, all features similar to PWS. However, chromosome 15q11-q13 methylation testing and fragile X studies were normal. GTG-banding and fluorescence in situ hybridization (FISH) with whole chromosome 3 paint probe (WCP3) and a chromosome 3p subtelomeric probe suggested a duplication of 3p25.3p26.2, a finding supported by comparative genomic hybridization (CGH). This region of chromosome 3p contains genes which contribute to obesity and behavioral problems, most notably, ghrelin (GHRL), an oxytocin receptor (OXTR), solute carrier family six members (gamma-aminobutyric acid (GABA) neurotransmitter transporters, SLC6A1 and SLC6A11), and peroxisome proliferator-activated receptor gamma (PPARG). To characterize these obesity and behavior related genes in our subject, we performed quantitative RT-PCR and compared expression levels with similarly aged male subjects (four non-obese males, four obese males, and four PWS males-two with 15q11-q13 deletions and two with maternal disomy 15). Our studies suggest increased expression of several genes in the 3p duplication region, including GHRL and PPARG, which may contribute to the phenotypic features in our 3p duplication subject.     

Splice-site variant in the RPS7 5′-UTR leads to a decrease in the mRNA level and development of Diamond-Blackfan anemia

Diamond-Blackfan anemia (DBA) is an inherited bone marrow failure syndrome characterized by erythroid aplasia. Pathogenic variants in ribosomal protein (RP) genes, GATA1, TSR2, and EPO, are considered to be the etiology of DBA. Variants in 5′-untranslated regions (UTRs) of these genes are poorly studied and can complicate the variant interpretation. We investigated the functional consequences NM_001011.4:c.-19 + 1G > T variant in the donor splice-site of the RPS7 5′-UTR. This variant was found in a family where two sons with DBA were carriers. Father, who also had this variant, developed myelodysplastic syndrome, which caused his death. Search for candidate causal variants and copy number variations in DBA-associated genes left RPS7 variant as the best candidate. Trio whole exome sequencing analysis revealed no pathogenic variants in other genes. Functional analysis using luciferase expression system revealed that this variant leads to disruption of splicing. Also, a decrease in the levels of mRNA and protein expression was detected. In conclusion, the established consequences of 5′-UTR splice-site variant c.-19 + 1G > T in the RPS7 gene provide evidence that it is likely pathogenic.     

Photocatalytic activity of layered MoS2 in the reductive degradation of bromophenol blue

Molybdenum disulphide (MoS₂) is a layered material with interesting photocatalytic properties. In this study, a layered MoS₂ was produced using a hydrothermal method. The obtained material was characterised by XRD (X-ray diffraction), XPS (X-ray photoelectron spectroscopy), SEM (scanning electron microscopy), UV-Vis spectroscopy, DLS (dynamic light scattering), and zeta potential analysis. For the evaluation of the photocatalytic properties of layered MoS₂, a solution of bromophenol blue (BPB) and the catalyst was illuminated for 120 minutes. According to the experimental results, MoS₂ exhibited excellent catalytic activity in BPB degradation. The MoS₂ preparation method enabled improved light harvesting, avoided fast charge recombination (related to bulk MoS₂), and created a large number of suitable electron transfer sites for photocatalytic reactions. Simulation of BPB decay and bromide production was carried out for a further understanding of MoS₂ photocatalytic action. The simulation results proved the reduction mechanism of BPB photodegradation.

Molybdenum disulphide (MoS₂) is a layered material with interesting photocatalytic properties.     

Structure and Spectral Luminescence Properties of (ZrO2)0.909(Y2O3)0.09(Eu2O3)0.001 Ceramics Synthesized by Uniaxial Compaction and Slip Casting

The structure, phase composition and spectral luminescence properties of single crystal and ceramic specimens of (ZrO₂)_0.909(Y₂O₃)_0.09(Eu₂O₃)_0.001 solid solutions synthesized using uniaxial compaction and slip casting techniques have been compared. The ceramic specimens have been synthesized from crushed single crystal specimens of similar composition. It has been shown that the crystalline structures of the ceramic and single crystal specimens are identical and cubic. The ceramic specimens synthesized using different methods prove to have close microstructure patterns. The spectral luminescence properties of Eu³⁺ ions in the (ZrO₂)_0.909(Y₂O₃)_0.09(Eu₂O₃)_0.001 ceramic specimens are similar to those of the single crystals with similar composition. The (ZrO₂)_0.909(Y₂O₃)_0.09(Eu₂O₃)_0.001 ceramic specimens prove to have uncontrolled Cr³⁺:Al₂O₃ impurities due to the synthesis conditions.     

Structural Formation of Soil Concretes Based on Loam and Fly Ash,Modified with a Stabilizing Polymer Additive

Finding new ways of recycling production waste to improve the characteristics of various building materials is an urgent scientific task. This article substantiates the possibility of the disposal of fly ash in the composition of soil concrete, which is used in the construction of the structural layers of road pavements, foundations of buildings and structures, as well as sites for various purposes. The scientific novelty lies in the fact that the structure formation of soil concretes based on loam and fly ash and modified with a stabilizing additive is being studied for the first time. It was found that the investigated fly ash, according to its hydraulic properties, is classified as latent active and can be introduced into the compositions of road soil concrete modified with additives of various resources. The effectiveness of the complex method of stabilization, due to changes in soil properties as a result of the use of the binding and stabilizing additives of polymer nature “Kriogelit”, is shown. It was found that the optimal content of binder and fly ash in the samples was 8 and 10 wt.%, respectively. It was established that the use of the stabilizing additive “Kriogelit” makes it possible to obtain soil concrete with the highest strength (compressive strength 2.5 MPa, flexural strength 0.5 MPa) and frost resistance of at least F15. The microstructure, the degree of dehydration and carbonization, and the phase composition of the initial raw mixtures and soil concretes stabilized with the addition of “Kriogelit” were studied by methods of scanning electron microscopy, X-ray diffraction analysis, differential scanning calorimetry, thermogravimetry, and infrared spectroscopy. It was shown that organo-mineral complexes, with the participation of polymer and montmorillonite, are formed in stabilized soil concrete. It was revealed that structure formation is accompanied by the physical adsorption of the polymer on active centers of silicate minerals, carbonization, and hydration–dehydration processes. It was found that the reason for the increase in the strength of stabilized soil concretes is the hydrophobization of the porous structure of minerals, as well as the formation of calcium oxide silicate and dicalcium hydrated silicate. By the method of performing biotests with the test objects Daphnia magna Straus and Chlorella vulgaris Beijer, it was proven that the developed road concretes modified with the stabilizing additive “Kriogelit” do not have an acute toxic effect on the test objects and are safe for the environment and human health.     

Visualization of the protective ability of a free radical trapping compound against rat C6 and F98 gliomas with diffusion tensor fiber tractography

PURPOSE: To apply fiber tractography to assess the effect of a possible antiglioma drug, phenyl N-tert-butyl nitrone (PBN), on glioma-affected neuronal fibers. The fiber tractography method was able to differentiate between different tumor types, such as the C6 and F98 rat glioma models. MATERIALS AND METHODS: C6 or F98 cells were intracranially injected into the cortex of male Fischer 344 rats. PBN treatment was initiated before or after cell implantation. Tumor growth was monitored with diffusion tensor imaging (DTI) and fiber tractography using diffusion-weighting gradients in 30 noncolinear directions. RESULTS: Although proton density-weighted (PDw) and T2-weighted (T2w) images did not show any difference between C6 and F98 gliomas without edema, the apparent diffusion coefficient (ADC) and fractional anisotropy (FA) maps were able to discriminate between these two tumor models. Fiber tractography was used to visualize C6 glioma-induced ischemia of tumor-surrounding tissues, whereas F98 glioma was found to infiltrate and penetrate into the corpus callosum (CC). During glioma growth, neuronal fibers were found to disappear at the border regions between the tumor and surrounding tissues. PBN treatment was shown to inhibit glioma growth with accompanying changes in the surrounding tissue. CONCLUSION: By noninvasively monitoring the degree of neuronal fiber integrity and connectivity with the use of neuronal fiber tractography, we were able to evaluate the protective effect of PBN against invasive glioma growth in rat brains. PBN provided protection of the neuronal fibers against tumor-induced ischemia and tumor invasion.     

Bioengineered Corneas Grafted as Alternatives to Human Donor Corneas in Three High‐Risk Patients

Corneas with severe pathologies have a high risk of rejection when conventionally grafted with human donor tissues. In this early observational study, we grafted bioengineered corneal implants made from recombinant human collagen and synthetic phosphorylcholine polymer into three patients for whom donor cornea transplantation carried a high risk of transplant failure. These patients suffered from corneal ulcers and recurrent erosions preoperatively. The implants provided relief from pain and discomfort, restored corneal integrity by promoting endogenous regeneration of corneal tissues, and improved vision in two of three patients. Such implants could in the future be alternatives to donor corneas for high‐risk patients, and therefore, merits further testing in a clinical trial.     

Amyloid accumulation is a late event in sporadic Alzheimer's disease-like pathology in nontransgenic rats

The amyloid cascade hypothesis posits that deposition of the amyloid β (Aβ) peptide in the brain is a key event in the initiation of Alzheimer''s disease (AD). Nonetheless, it now seems increasingly unlikely that amyloid toxicity is the cause of sporadic AD, which leads to cognitive decline. Here, using accelerated-senescence nontransgenic OXYS rats, we confirmed that aggregation of Aβ is a later event in AD-like pathology. We showed that an age-dependent increase in the levels of Aβ_1–42 and extracellular Aβ deposits in the brain of OXYS rats occur later than do synaptic losses, neuronal cell death, mitochondrial structural abnormalities, and hyperphosphorylation of the tau protein. We identified the variants of the genes that are strongly associated with the risk of either late-onset or early-onset AD, including App, Apoe4, Bace1, Psen1, Psen2, and Picalm. We found that in OXYS rats nonsynonymous SNPs were located only in the genes Casp3 and Sorl1. Thus, we present proof that OXYS rats may be a model of sporadic AD. It is possible that multiple age-associated pathological processes may precede the toxic amyloid accumulation, which in turn triggers the final stage of the sporadic form of AD and becomes a hallmark event of the disease.