Detection of haemoglobin S using multiplex ligation-dependent probe amplification and flow-through hybridization techniques: experience in a tertiary hospital

Haemoglobin S (HbS, α2β26GluVal) is a variant haemoglobin resulted from GAGGTG mutation on codon 6 of HBB gene. HbS haemoglobinopathy is uncommon in Malaysia and mainly seen in immigrants. However, Malaysian Indians and Malays are rarely affected. This study reviewed the laboratory findings of patients with HbS and the utilization of multiplex ligation-dependent probe amplification (MLPA) and flow-through hybridization (FTH) in the detection of HbS mutation. HbS was identified and quantified by high performance liquid chromatography (HPLC), capillary electrophoresis (CE) and cellulose acetate gel electrophoresis. Molecular analysis was performed using MLPA, FTH and Sanger Sequencing. Two Africans, three Malays and two Indian individuals aged between 2-31 years were identified from our laboratory. Five patients were homozygous HbS, one was compound heterozygous HbS/β-thalassemia and one was a carrier of HbS. The patients with homozygous HbS had their haemoglobin (Hb) ranging from 7.4-10.2 g/dL with HbS and HbF levels of 58.3-94.7% and 1.5-35.5%, respectively. The Hb of compound heterozygous HbS/β-thalassaemia patients was 5.8 g/dL and was normal in heterozygous HbS. HbS, HbF and HbA2 levels for the HbS/β-thalassaemia and the carrier were 67%, 27.2% and 4.2%, and 38.6%, 0.1% and 2.8%, respectively. Both MLPA and FTH successfully detected HbS mutation in all cases but only FTH could identify the zygosity of the HbS mutation together with underlying concomitant β-thalassaemia in a single test

Drawing from Grotowski and Beyond: Kuo Pao Kun’s Discourse on Audiences in Singapore in the 1980s

Much has been researched on Kuo Pao Kun’s multilingualism and multiculturalism. However, as one of one of the most important Asian dramatists, the analysis of Kuo’s discourse on audiences remains largely unexplored. There is a pressing need to understand the ways which theatre practitioners imagine audiences as it points to issues of subjectivity, audience participation and social engagement, especially in a neoliberal society like Singapore where people are often positioned as docile economic subjects. Among the many Asian and Western dramatists Kuo drew inspiration from, Jerzy Grotowski was pivotal. This essay seeks to address this gap by examining how the latter’s ideas was crucial to understanding how Kuo envisioned theatre and audiences alongside his artistic practice

HNRNPM controls circRNA biogenesis and splicing fidelity to sustain cancer cell fitness

High spliceosome activity is a dependency for cancer cells, making them more vulnerable to perturbation of the splicing machinery compared to normal cells. To identify splicing factors important for prostate cancer (PCa) fitness, we performed pooled shRNA screens in vitro and in vivo. Our screens identified HNRNPM as a regulator of PCa cell growth. RNA- and eCLIP-sequencing identified HNRNPM binding to transcripts of key homeostatic genes. HNRNPM binding to its targets prevents aberrant exon inclusion and back-splicing events. In both linear and circular mis-spliced transcripts, HNRNPM preferentially binds to GU-rich elements in long flanking proximal introns. Mimicry of HNRNPM dependent linear splicing events using splice-switching-antisense-oligonucleotides (SSOs) was sufficient to inhibit PCa cell growth. This suggests that PCa dependence on HNRNPM is likely a result of mis-splicing of key homeostatic coding and non-coding genes. Our results have further been confirmed in other solid tumors. Taken together, our data reveal a role for HNRNPM in supporting cancer cell fitness. Inhibition of HNRNPM activity is therefore a potential therapeutic strategy in suppressing growth of PCa and other solid tumors

A community-based validation study of the short-form 36 version 2 philippines (tagalog) in two cities in the philippines

10.1371/journal.pone.0083794PLoS ONE812-POLN

Comparison of digital and conventional impression techniques: evaluation of patients’ perception, treatment comfort, effectiveness and clinical outcomes

Background: The purpose of this study was to compare two impression techniques from the perspective of patient preferences and treatment comfort.Methods: Twenty-four (12 male, 12 female) subjects who had no previous experience with either conventional or digital impression participated in this study. Conventional impressions of maxillary and mandibular dental arches were taken with a polyether impression material (Impregum, 3 M ESPE), and bite registrations were made with polysiloxane bite registration material (Futar D, Kettenbach). Two weeks later, digital impressions and bite scans were performed using an intra-oral scanner (CEREC Omnicam, Sirona). Immediately after the impressions were made, the subjects' attitudes, preferences and perceptions towards impression techniques were evaluated using a standardized questionnaire. The perceived source of stress was evaluated using the State-Trait Anxiety Scale. Processing steps of the impression techniques (tray selection, working time etc.) were recorded in seconds. Statistical analyses were performed with the Wilcoxon Rank test, and p < 0.05 was considered significant.Results: There were significant differences among the groups (p < 0.05) in terms of total working time and processing steps. Patients stated that digital impressions were more comfortable than conventional techniques.Conclusions: Digital impressions resulted in a more time-efficient technique than conventional impressions. Patients preferred the digital impression technique rather than conventional techniques

Ethnic differences in the association between blood pressure components and chronic kidney disease in middle aged and older Asian adults

10.1186/1471-2369-14-86BMC Nephrology141

Self-oligomerization regulates stability of survival motor neuron protein isoforms by sequestering an SCF^Slmb degron

Spinal muscular atrophy (SMA) is caused by homozygous mutations in human SMN1. Expression of a duplicate gene (SMN2) primarily results in skipping of exon 7 and production of an unstable protein isoform, SMNΔ7. Although SMN2 exon skipping is the principal contributor to SMA severity, mechanisms governing stability of survival motor neuron (SMN) isoforms are poorly understood. We used a Drosophila model system and label-free proteomics to identify the SCFSlmb ubiquitin E3 ligase complex as a novel SMN binding partner. SCFSlmb interacts with a phosphor degron embedded within the human and fruitfly SMN YG-box oligomerization domains. Substitution of a conserved serine (S270A) interferes with SCFSlmb binding and stabilizes SMNΔ7. SMA-causing missense mutations that block multimerization of full-length SMN are also stabilized in the degron mutant background. Overexpression of SMNΔ7S270A, but not wild-type (WT) SMNΔ7, provides a protective effect in SMA model mice and human motor neuron cell culture systems. Our findings support a model wherein the degron is exposed when SMN is monomeric and sequestered when SMN forms higher-order multimers