Protein Kinase C inhibitor Molecule Effect Partial Reversal of Spliceopathy in Myotonic Dystrophy Type 2 Cell Line

Myotonic dystrophy is a neuromuscular disease which manifest as two forms namely type 1 (DM1) and type 2 (DM2). One of the major molecular events associated with the disease condition is misregulation of splicing resulting in spliceopathy. This study was performed to assess the effect of protein kinase C (PKC) inhibitors- Ro 31-8220 and hypericin- on the splicing of insulin receptor (IR) and muscleblind-like (MBNL1) in myotonic dystrophy type 2 (DM2) cell line. The results of this study demonstrated that only Ro 31-8820 was able to effect partial rescue of missplicing of insulin receptor and muscleblind-like 1. It indicates that it could be a possible therapeutic agent for treatment of DM2. Keywords: Myotonic dystrophy, Protein Kinase C inhibitor, spliceopathy, therapeutics DOI: 10.7176/JNSR/11-14-03 Publication date:July 31st 202

Non-Association of Toll-Like Receptor 2 rs3804099 Polymorphism with Paediatric Tuberculosis in South Africa

Genetic polymorphisms are important risk factors that are involved in development and severity of disease infections especially tuberculosis. In this study, a non-synonymous single nucleotide polymorphism of immune response protein gene, toll-like receptor 2 (TLR2) occurring in exon 2 (rs3804099) was carried out by genotypic analysis of cases and controls for tuberculosis. The paediatric study population consisted of 151 cases and 82 controls obtained mainly from Xhosa ethnic group.Result from this study showed that this non-synonymous single nucleotide polymorphism of TLR2 was not associated with tuberculosis (p= 0.38). Keywords: Tuberculosis, Toll-like Receptor 2, Paediatrics, Single Nucleotide Polymorphisms DOI: 10.7176/ALST/74-06 Publication date:May 31st 201

The Occurrence of High Frequencies of MBL2 polymorphisms in the Xhosa ethnic group of South Africa

Polymorphic changes in a given population give rise to genetic variations. These variations account for differences found within a population. One of the most abundant and common genetic variant that occur in the population is the Single Nucleotide Polymorphisms which are point mutations in genes. The study assessed the frequency of single nucleotide polymorphisms in mannose binding lectin (MBL2) gene namely rs8191996, rs1838065, rs8179079 and rs1800450 in 366 adult individuals drawn from Xhosa ethnic group of South Africa. The polymorphisms showed high frequencies in its occurrence which was 0.71, 0.42, 0.71 and 0.73 for rs8191996, rs1838065, rs1800450 and rs8179079 respectively. Additionally, the genotype distribution of these polymorphisms showed a predominance of homozygotes over heterozygotes and were in accordance with Hardy-Weinberg principle. The high frequencies of the MBL2 polymorphisms under study in this population suggest that there is a high incidence of single gene mutations in Xhosa ethnic group which could make the population vulnerable to infection. DOI: 10.7176/JBAH/10-6-04 Publication date:March 31st 202

Association of MBL2 Codon 34 Variant of Exon 1 with Severity of Tuberculosis in Paediatric Population of South Africa

The host genetic factors contribute to determining the susceptibility of an individual to an infectious disease and this forms the basis of this study. Many gene polymorphisms have been implicated in disease development and progression.In this study, a genotypic case-control study was carried out to determine the effect of single nucleotide polymorphisms in MBL2 on tuberculosis on paediatrics drawn mainly from the Xhosa ethnic group, genotyped together with 366 adult controls from Xhosa population. The MBL2 non-synonymous SNP at codon 34 of exon 1 was found to be associated with the development of TB in the study population especially Xhosa ethnic group (p =0.00002).Therefore, codon 34 variant allele of MBL2 may be a risk factor in the development of TB in paediatric population. Keywords: Tuberculosis, MBL2, Single Nucleotide Polymorphisms, Paediatric

Mannose Binding Lectin Levels Was Not Associated with Resistance to Tuberculosis Infection in the Population of Uyo Metropolis in Nigeria

Mannose-binding lectin (MBL2) is an important pattern recognition molecule that identifies and binds to specific sugar molecules on the surface of pathogens thereby activating its destruction by the immune system. Samples for study were recruited from Uyo metropolis of Akwa Ibom state in Nigeria. In this study, levels of MBL2 was measured by enzyme-linked immunosorbent assay in tuberculosis patients and healthy individuals to determine if the immune protein protects against tuberculosis infection. MBL2 levels in tuberculosis patients and healthy controls were 14.0ng/ml ± 13.9 and 19.9ng/ml ± 18.5 respectively. The results from the study showed that there was no association in MBL2 levels between tuberculosis and controls (p=0.107) as well as between the different sub-groups. Therefore, MBL2 is not a contributory factor in resistance against tuberculosis in the population under study. Keywords: Mannose binding lectin, tuberculosis, pattern recognition molecule, immune system DOI: 10.7176/JNSR/12-14-02 Publication date:July 31st 202

A Cell-Based Double Reporter Gene Splicing Assay for Therapeutic Screening in Myotonic Dystrophy

The study has developed a model splicing construct assay system based on splicing misregulation, one of the major molecular features associated with myotonic dystrophy. The splicing construct assay has double reporters for intron 2 splicing in chloride channel (CLCN1). The CLCN1 transgene splicing construct assay was used to transfect wild type and DM fibroblast cell lines and the clones generated showed that it enabled quantification of splicing efficiency in transgene construct. Validation of the DM fibroblasts containing transgene splicing construct was performed by differentiating the DM fibroblasts into myoblasts which exhibited a switch in CLCN1 splicing construct which was consistent with that associated with myotonic dystrophy (DM) condition. The myoblast derived from fibroblasts cell-based gene-splicing assay was subsequently applied in therapeutic screening in small throughput screens of 113 compounds which identified Protein Kinase C inhibitors- hypericin and Ro-31-8220 as potential therapeutic agents. The CLCN1 gene-splicing assay is a good model system for application in therapeutic screening in myotonic dystrophy because its double reporters facilitated quantification of effect putative drug on correction of misregulated splicing

Adaptation of the Wound Healing Questionnaire universal-reporter outcome measure for use in global surgery trials (TALON-1 study): mixed-methods study and Rasch analysis

BackgroundThe Bluebelle Wound Healing Questionnaire (WHQ) is a universal-reporter outcome measure developed in the UK for remote detection of surgical-site infection after abdominal surgery. This study aimed to explore cross-cultural equivalence, acceptability, and content validity of the WHQ for use across low- and middle-income countries, and to make recommendations for its adaptation.MethodsThis was a mixed-methods study within a trial (SWAT) embedded in an international randomized trial, conducted according to best practice guidelines, and co-produced with community and patient partners (TALON-1). Structured interviews and focus groups were used to gather data regarding cross-cultural, cross-contextual equivalence of the individual items and scale, and conduct a translatability assessment. Translation was completed into five languages in accordance with Mapi recommendations. Next, data from a prospective cohort (SWAT) were interpreted using Rasch analysis to explore scaling and measurement properties of the WHQ. Finally, qualitative and quantitative data were triangulated using a modified, exploratory, instrumental design model.ResultsIn the qualitative phase, 10 structured interviews and six focus groups took place with a total of 47 investigators across six countries. Themes related to comprehension, response mapping, retrieval, and judgement were identified with rich cross-cultural insights. In the quantitative phase, an exploratory Rasch model was fitted to data from 537 patients (369 excluding extremes). Owing to the number of extreme (floor) values, the overall level of power was low. The single WHQ scale satisfied tests of unidimensionality, indicating validity of the ordinal total WHQ score. There was significant overall model misfit of five items (5, 9, 14, 15, 16) and local dependency in 11 item pairs. The person separation index was estimated as 0.48 suggesting weak discrimination between classes, whereas Cronbach's α was high at 0.86. Triangulation of qualitative data with the Rasch analysis supported recommendations for cross-cultural adaptation of the WHQ items 1 (redness), 3 (clear fluid), 7 (deep wound opening), 10 (pain), 11 (fever), 15 (antibiotics), 16 (debridement), 18 (drainage), and 19 (reoperation). Changes to three item response categories (1, not at all; 2, a little; 3, a lot) were adopted for symptom items 1 to 10, and two categories (0, no; 1, yes) for item 11 (fever).ConclusionThis study made recommendations for cross-cultural adaptation of the WHQ for use in global surgical research and practice, using co-produced mixed-methods data from three continents. Translations are now available for implementation into remote wound assessment pathways