Genotype-Phenotype Correlations in PMM2-CDG

PMM2-CDG is a rare disease, causing hypoglycosylation of multiple proteins, hence preventing full functionality. So far, no direct genotype–phenotype correlations have been identified. We carried out a retrospective cohort study on 26 PMM2-CDG patients. We collected the identified genotype, as well as continuous variables indicating the disease severity (based on Nijmegen Pediatric CDG Rating Score or NPCRS) and dichotomous variables reflecting the patients’ phenotype. The phenotypic effects of patients’ genotype were studied using non-parametric and Chi-Square tests. Seventeen different pathogenic variants have been studied. Variants with zero enzyme activity had no significant impact on the Nijmegen score. Pathogenic variants involving the stabilization/folding domain have a significantly lower total NPCRS (p = 0.017): presence of the p.Cys241Ser mutation had a significantly lower subscore 1,3 and NPCRS (p = 0.04) and thus result in a less severe phenotype. On the other hand, variants involving the dimerization domain, p.Pro113Leu and p.Phe119Leu, resulted in a significantly higher NPCRS score (p = 0.002), which indicates a worse clinical course. These concepts give a better insight in the phenotypic prognosis of PMM2-CDG, according to their molecular base.

Molecular Proteomics and Signalling of Human Platelets in Health and Disease

Platelets are small anucleate blood cells that play vital roles in haemostasis and thrombosis, besides other physiological and pathophysiological processes. These roles are tightly regulated by a complex network of signalling pathways. Mass spectrometry-based proteomic techniques are contributing not only to the identification and quantification of new platelet proteins, but also reveal post-translational modifications of these molecules, such as acetylation, glycosylation and phosphorylation. Moreover, target proteomic analysis of platelets can provide molecular biomarkers for genetic aberrations with established or non-established links to platelet dysfunctions. In this report, we review 67 reports regarding platelet proteomic analysis and signalling on a molecular base. Collectively, these provide detailed insight into the: (i) technical developments and limitations of the assessment of platelet (sub)proteomes; (ii) molecular protein changes upon ageing of platelets; (iii) complexity of platelet signalling pathways and functions in response to collagen, rhodocytin, thrombin, thromboxane A2 and ADP; (iv) proteomic effects of endothelial-derived mediators such as prostacyclin and the anti-platelet drug aspirin; and (v) molecular protein changes in platelets from patients with congenital disorders or cardiovascular disease. However, sample sizes are still low and the roles of differentially expressed proteins are often unknown. Based on the practical and technical possibilities and limitations, we provide a perspective for further improvements of the platelet proteomic field.

Dynamic association of IκBα to chromatin is regulated by acetylation and cleavage of histone H4

ABSTRACTIκBs exert a principal function as cytoplasmic inhibitors of the NF-kB transcription factors. Additional functions for IκB homologues have been described including association to chromatin and transcriptional regulatioin. Phosphorylated and SUMOylated IκBα (pS-IκBα) binds histones H2A and H4 in the stem and progenitor compartment of skin and intestine, but the mechanisms controlling its recruitment to chromatin are largely unstudied.We here show that serine 32-36 phosphorylation of IκBα favors its binding with nucleosomes and demonstrated that p-IκBα association to H4 is favored by acetylation at specific H4 lysine residues. N-terminal tail of H4 is lost during intestinal cell differentiation by proteolytic cleavage at residues 17-19 imposed ny trypsin or chymotrypsin, which interferes p-IκBα binding. Paradoxically, inhibition of trypsin and chymotrypsin activity in HT29 cells increased p-IκBα chromatin binding and impaired goblet cell differentiation, comparable to IκBα deletion. Together our results indicate that dynamic binding of IκBα to chromatin is a requirement for intestinal cell differentiation and provide a molecular base for the restricted nuclear distribution of p-IκBα at specific stem cell compartments.

Determination of mutation in the coding regions of FAM83H and ENAM genes in patients with imperfect enamel (Amelogenesis Imperfecta)

Introduction: Tooth enamel is a precious and highly mineralized tissue in the human body. Amelogenesis Imperfecta (AI) is a developmental, evolutionary and hereditary disease presents with the rare abnormal formation of enamel that affects the primary and secondary dentition. The molecular base of the incomplete quinizium together with clinical manifestation suggest that AI may result from mutations in the FAM83H and ENAM genes. In this study, we aimed to evalu- ate the association between Amelogenesis Imperfecta and mutations in the FAM83H and ENAM genes in 18 Iranian families with AI in dominant and non-syndromic form. Materials and Methods: 18 Iranian families with at least 1 patient with Amelogenesis Imperfecta were included in this case study and were examined for related specific manifestations and also, 10CC of blood was taken from each patient followed by PCR and genome sequence for genetic alterations in FAM83H and ENAM. Genome sequences were analyzed using CLC software and CLC Sequence Viewer was used to compare them with reference sequences in the RefSeq database at the NCBI later were discussed together with clinical manifestations for each patient. Results: All patients showed a mutation in the exon 5 of FAM83H gene in nucleotide rs56148058C/T which converted Serotonin to Aspartin. In two patients carried a mutation in the nucleotide rs546809055A/G that changed Leucine to Phenylalanine. None of patients showed sig- nificant alteration in the ENAM gene. Conclusion: This study indicates that FAM83H gene plays an import role in incidence of Amelogenesis Imperfecta in Iran.

First rDNA sequence data for Haplosplanchnus pachysomus (Digenea: Haplosplanchnidae) ex Mugil cephalus from the Black Sea, and molecular evidence for cryptic species within Haplosplanchnus pachysomus (Digenea: Haplosplanchnidae) in Palaearctic and Indo-West Pacific regions

Adult trematodes, morphologically similar to Haplosplanchnus pachysomus (Eysenhardt, 1829), were extracted from the intestine of Mugil cephalus Linnaeus, 1758, collected in the Black Sea basin. Morphological, morphometric and 28S ribosomal DNA (rDNA) partial sequence data were obtained for these trematodes following comparative analysis with previous data on this species. Worms from this study were morphologically identical to all previously reported H. pachysomus specimens from different locations. The results of the morphometric analysis indicated general similarity between H. pachysomus from the Black Sea and trematodes from Vietnam and Australia. Trematodes from the Black Sea and specimens from Spain were identical based on 28S rDNA partial sequences; however, these sequences differed from that of H. pachysomus from Vietnam and Australia by eight fixed substitutions. Overall, our results indicate that H. pachysomus from Spain and the Black Sea and from Vietnam and Australia can presently be considered as two cryptic species, one in Palaearctic and one in Indo-West Pacific regions. Our results provide a molecular base for including Haplosplanchnus purii in the genus Provitellotrema or to consider H. purii, P. crenimugilis and H. pachysomus within the same genus.

IDENTIFICATION LOCAL ISOLATES OF Trichophyton mentagrophytes AND DETECTION OF KERATINASE GENE USING PCR TECHNIQUE

This study was aimed to identify dermatophytic selective isolate using PCR technique as a rapid molecular assay. The results of this study showed among 60 samples of patients suffering from ringworm disease. forty isolates (66%) were Trichophyton mentagrophytes which diagnosed as dermatophytosis according to morphological and cultural methods. In order to investigate the ability of isolates to keratin analyses using solid medium supplemented with keratin azure , the results revealed that 20 isolates appeared best ability to keratin analysis and nine isolates had best ability for keratinase production in submerged culture. According to this results T. mantagrophytes (K3) (had higher activity for keratinase) was chosen for molecular identification. The results of PCR revealed that primer for18S rRNA gene of T. mentagrophytes K1 isolate and specific primer for subtilisin like protease gene were amplified and appeared as single DNA band with a molecular base of 690 bp and 623bp respectively .The blast result of sample sequences of amplified fragment revealed that the isolate were 100% identical to reference sequence of T.mentagrophytes var. interdigtal and depending on data base in NCBI The result of PCR product for enzyme showed new type named GBF60362 (402) subtilisin-like protease related to T. mentagrophytes 1354684064 BFBSOLP00892.

Circular RNAs: A New Piece in the Colorectal Cancer Puzzle

Colorectal cancer (CRC) is the third most fatal type of malignancy, worldwide. Despite the advances accomplished in the elucidation of its molecular base and the existing CRC biomarkers introduced in the clinical practice, additional research is required. Circular RNAs (circRNAs) constitute a new RNA type, formed by back-splicing of primary transcripts. They have been discovered during the 1970s but were characterized as by-products of aberrant splicing. However, the modern high-throughput approaches uncovered their widespread expression; therefore, several questions were raised regarding their potential biological roles. During the last years, great progress has been achieved in the elucidation of their functions: circRNAs can act as microRNA sponges, transcription regulators, and interfere with splicing, as well. Furthermore, they are heavily involved in various human pathological states, including cancer, and could serve as diagnostic and prognostic biomarkers in several diseases. Particularly in CRC, aberrant expression of circRNAs has been observed. More specifically, these molecules either inhibit or promote colorectal carcinogenesis by regulating different molecules and signaling pathways. The present review discusses the characteristics and functions of circRNA, prior to analyzing the multifaceted role of these molecules in CRC and their potential value as biomarkers and therapeutic targets.

Genetic Base of Behavioral Disorders in Mucopolysaccharidoses: Transcriptomic Studies

Mucopolysaccharidoses (MPS) are a group of inherited metabolic diseases caused by mutations leading to defective degradation of glycosaminoglycans (GAGs) and their accumulation in cells. Among 11 known types and subtypes of MPS, neuronopathy occurs in seven (MPS I, II, IIIA, IIIB, IIIC, IIID, VII). Brain dysfunctions, occurring in these seven types/subtypes include various behavioral disorders. Intriguingly, behavioral symptoms are significantly different between patients suffering from various MPS types. Molecular base of such differences remains unknown. Here, we asked if expression of genes considered as connected to behavior (based on Gene Ontology, GO terms) is changed in MPS. Using cell lines of all MPS types, we have performed transcriptomic (RNA-seq) studies and assessed expression of genes involved in behavior. We found significant differences between MPS types in this regard, with the most severe changes in MPS IIIA (the type considered as the behaviorally most severely affected), while the lowest changes in MPS IVA and MPS VI (types in which little or no behavioral disorders are known). Intriguingly, relatively severe changes were found also in MPS IVB (in which, despite no behavioral disorder noted, the same gene is mutated as in GM1 gangliosidosis, a severe neurodegenerative disease) and MPS IX (in which only a few patients were described to date, thus, behavioral problems are not well recognized). More detailed analyses of expression of certain genes allowed us to propose an association of specific changes in the levels of transcripts in specific MPS types to certain behavioral disorders observed in patients. Therefore, this work provides a principle for further studies on the molecular mechanism of behavioral changes occurring in MPS patients.