A Study on the Level of Acceptance towards the Islamic Home Financing: Empirical Study in Malaysia

This paper offers a conceptual understanding of the Islamic housing loan (BaiBithaman Ajil) with a special focus on the operational matters and the factors that affect the acceptance of Bai-Bithaman Ajil. Approach: This paper incorporates relevant findings and issues surrounding the acceptance and implementation of the Islamic housing loan Findings: This paper offer more insights on the challenges faced by Islamic banking institutions to deal with both muslim and non-muslim markets in terms of the marketability of the product especially multi-racial society

Evaluation of the SpeeDx ResistancePlus® GC and SpeeDx GC 23S 2611 (beta) molecular assays for prediction of antimicrobial resistance/susceptibility to ciprofloxacin and azithromycin in Neisseria gonorrhoeae

European collaborative group: Raquel Abad Torreblanca, Lena Rós Ásmundsdóttir, Eszter Balla, Irith De Baetselier, Beatrice Bercot, Thea Bergheim, Maria José Borrego, Susanne Buder, Robert Cassar, Michelle Cole, Alje van Dam, Claudia Eder, Steen Hoffmann, Blazenka Hunjak, Samo Jeverica, Vesa Kirjavainen, Panayiota Maikanti-Charalambous, Vivi Miriagou, Beata Młynarczyk-Bonikowska, Gatis Pakarna, Peter Pavlik, Monique Perrin, Joseph Pett, Paola Stefanelli, Kate Templeton, Magnus Unemo, Jelena Viktorova, Hana ZákouckáPortugal: Maria-José Borrego (INSA)Background: Accurate molecular assays for prediction of antimicrobial resistance (AMR)/susceptibility in Neisseria gonorrhoeae (Ng) can offer individualized treatment of gonorrhoea and enhanced AMR surveillance. Objectives: We evaluated the new ResistancePlus® GC assay and the GC 23S 2611 (beta) assay (SpeeDx), for prediction of resistance/susceptibility to ciprofloxacin and azithromycin, respectively. Methods: Nine hundred and sixty-seven whole-genome-sequenced Ng isolates from 20 European countries, 143 Ng-positive (37 with paired Ng isolates) and 167 Ng-negative clinical Aptima Combo 2 (AC2) samples, and 143 non-gonococcal Neisseria isolates and closely related species were examined with both SpeeDx assays. Results: The sensitivity and specificity of the ResistancePlus® GC assay to detect Ng in AC2 samples were 98.6% and 100%, respectively. ResistancePlus® GC showed 100% sensitivity and specificity for GyrA S91 WT/S91F detection and 99.8% sensitivity and specificity in predicting phenotypic ciprofloxacin resistance. The sensitivity and specificity of the GC 23S 2611 (beta) assay for Ng detection in AC2 samples were 95.8% and 100%, respectively. GC 23S 2611 (beta) showed 100% sensitivity and 99.9% specificity for 23S rRNA C2611 WT/C2611T detection and 64.3% sensitivity and 99.9% specificity for predicting phenotypic azithromycin resistance. Cross-reactions with non-gonococcal Neisseria species were observed with both assays, but the analysis software solved most cross-reactions. Conclusions: The new SpeeDx ResistancePlus® GC assay performed well in the detection of Ng and AMR determinants, especially in urogenital samples. The GC 23S 2611 (beta) assay performed relatively well, but its sensitivity, especially for predicting phenotypic azithromycin resistance, was suboptimal and further optimizations are required, including detection of additional macrolide resistance determinant(s).This work was supported by the O¨rebro County Council Research Committee and the Foundation for Medical Research at O¨rebro University Hospital, O¨rebro, Sweden.info:eu-repo/semantics/publishedVersio

Expression proteomics of UPF1 knockdown in HeLa cells reveals autoregulation of hnRNP A2/B1 mediated by alternative splicing resulting in nonsense-mediated mRNA decay

BACKGROUND: In addition to acting as an RNA quality control pathway, nonsense-mediated mRNA decay (NMD) plays roles in regulating normal gene expression. In particular, the extent to which alternative splicing is coupled to NMD and the roles of NMD in regulating uORF containing transcripts have been a matter of debate. RESULTS: In order to achieve a greater understanding of NMD regulated gene expression we used 2D-DiGE proteomics technology to examine the changes in protein expression induced in HeLa cells by UPF1 knockdown. QPCR based validation of the corresponding mRNAs, in response to both UPF1 knockdown and cycloheximide treatment, identified 17 bona fide NMD targets. Most of these were associated with bioinformatically predicted NMD activating features, predominantly upstream open reading frames (uORFs). Strikingly, however, the majority of transcripts up-regulated by UPF1 knockdown were either insensitive to, or even down-regulated by, cycloheximide treatment. Furthermore, the mRNA abundance of several down-regulated proteins failed to change upon UPF1 knockdown, indicating that UPF1`s role in regulating mRNA and protein abundance is more complex than previously appreciated. Among the bona fide NMD targets, we identified a highly conserved AS-NMD event within the 3` UTR of the HNRNPA2B1 gene. Overexpression of GFP tagged hnRNP A2 resulted in a decrease in endogenous hnRNP A2 and B1 mRNA with a concurrent increase in the NMD sensitive isoforms. CONCLUSIONS: Despite the large number of changes in protein expression upon UPF1 knockdown, a relatively small fraction of them can be directly attributed to the action of NMD on the corresponding mRNA. From amongst these we have identified a conserved AS-NMD event within HNRNPA2B1 that appears to mediate autoregulation of HNRNPA2B1 expression levels

A Pilot Study to Non-Invasively Track PIK3CA Mutation in Head and Neck Cancer

Background: PIK3CA pathways are the most frequently mutated oncogenic pathway in head and neck squamous cell carcinoma (HNSCC), including virally driven HNCs. PIK3CA is involved in the PI3K-PTEN-mTOR signalling pathway. PIK3CA has been implicated in HNSCC progression and PIK3CA mutations may serve as predictive biomarkers for therapy selection. Circulating tumour DNA (ctDNA) derived from necrotic and apoptotic tumour cells are thought to harbour tumour-specific genetic alterations. As such, the detection of PIK3CA alterations detected by ctDNA holds promise as a potential biomarker in HNSCC. Methods: Blood samples from treatment naïve HNSCC patients (n = 29) were interrogated for a commonly mutated PIK3CA hotspot mutation using low cost allele-specific Plex-PCRTM technology. Results: In this pilot, cross sectional study, PIK3CA E545K mutation was detected in the plasma samples of 9/29 HNSCC patients using the Plex-PCRTM technology. Conclusion: The results of this pilot study support the notion of using allele-specific technologies for cost-effective testing of ctDNA, and further assert the potential utility of ctDNA in HNSCC

Evaluation of the ResistancePlus GC (beta) assay: A commercial diagnostic test for the direct detection of ciprofloxacin susceptibility or resistance in Neisseria gonorrhoeae

Objectives: To evaluate the performance of the ResistancePlus GC (beta) assay for the simultaneous detection of Neisseria gonorrhoeae and gyrA S91 markers of resistance (S91F) and susceptibility (WT) to ciprofloxacin, from both clinical specimens and isolates. Methods: Performance was assessed on several sample banks, including N. gonorrhoeae isolates (n"822), nongonococcal isolates (n"110), N. gonorrhoeae-positive clinical specimens (n"402) and N. gonorrhoeae-negative specimens (n"290). Results were compared with previous testing data, including S91 genotyping and phenotypic resistance profiles. Results: Overall, the assay demonstrated 100% sensitivity for N. gonorrhoeae detection in clinical isolates. For gyrA S91 mutation detection in clinical isolates, the assay showed 100% sensitivity/specificity compared with the genotype, and .99%/>97% sensitivity/specificity when compared with phenotype. For positive clinical specimens, the assay demonstrated>96% sensitivity for N. gonorrhoeae detection and 100% sensitivity/specificity for gyrA S91 mutation detection. The assay demonstrated>99% specificity for N. gonorrhoeae detection against non-gonococcal isolates and 100% specificity for negative clinical specimens. Conclusions: The ResistancePlus GC (beta) assay is suitable for the detection of N. gonorrhoeae and gyrA markers associated with resistance/susceptibility to ciprofloxacin directly in clinical samples. This assay could be implemented for the individualized treatment of gonorrhoea infections as well as to enhance current antimicrobial resistance surveillance methods

Superior Multiplexing Capacity of PlexPrimers Enables Sensitive and Specific Detection of SNPs and Clustered Mutations in qPCR

<div><p>Background</p><p>Whilst qPCR provides an extremely powerful tool for genetic analysis, some applications such as multiplexing variant alleles (eg SNPs, point mutations or deletions), remain challenging using current primer/probe systems. The novel design features of PlexPrimers allow sensitive, multiplexed analysis of variant alleles even when these are tightly clustered.</p><p>Method</p><p>PlexPrimers were combined with PlexZymes in qPCR assays for the detection of SNPs in human absorption, distribution, metabolism, and excretion (ADME) genes; clustered mutations in the 23S rRNA gene which confer antibiotic resistance to <i>Mycoplasma genitalium</i>; and deletions within the human epidermal growth factor receptor (EGFR) gene.</p><p>Results</p><p>The combination of PlexPrimers and PlexZymes allowed robust multiplexing of targets which resulted in 100% concordance with results obtained using hydrolysis probe kits for 14 SNPs in the ADME genes. A 7-plex qPCR assay targeting <i>M</i>. <i>genitalium</i>, 5 clustered mutations associated with macrolide resistance and an internal control, allowed efficient amplification of all targets, with all 5 mutations detected in a single channel. Finally, the strategy was employed to analyse common EGFR mutants with high sensitivity, detecting deletions present at only 0.01%.</p><p>Conclusion</p><p>PlexPrime is a novel technology for the detection of genetic variants. Unlike previous strategies, the combination of PlexPrimers with PlexZymes enables both allele-specific detection and allele-specific amplification in qPCR. The study demonstrated highly sensitive and specific detection of mutations and SNPs, and superior multiplexing capacity. The ability to multiplex clustered genetic variants reduces the time to result providing more actionable information.</p></div

Comparison of PlexPrime/PlexZyme and hydrolysis probe assays for genotyping ADME SNPs showing real-time amplification plots and allelic discrimination graphs.

<p>(A) The PlexPrime/PlexZyme 1-well triplex assay reads C, A and T results in the Quasar 705, FAM and Texas Red channels, respectively. (B) The hydrolysis probe 2-well assay reads C and A in the VIC and FAM channels in well 1, and reads C and T in the VIC and FAM channels in well 2. The three samples used in these exemplary graphs are (i) 18855 which is homozygous for the C allele, (ii) 18608 which is homozygous for the A allele and (iii) 18970 which is heterozygous for alleles A and T. The Cq values for each allelic assay are shown on the amplification plot and the plotted graph for the C assay is orange, the A assay is blue and the T assay is green. The allelic discrimination plots display all of the samples listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0170087#pone.0170087.t001" target="_blank">Table 1</a> for the SNP ABCB1 rs2032582; where (C) are the PlexPrime/PlexZyme plots derived from a single well and (D) are the Hydrolysis probe plots derived from two wells. The allelic discrimination plots have either a) Allele 1 in orange (C/C or C/A), Allele 2 in blue (T/T or A/T), Heterozygous in green (C/T) or None in black (NTC or A/A); or b) Allele 1 in orange (C/C or C/T), Allele 2 in blue (A/A or A/T), Heterozygous in green (C/A) or None in black (NTC or T/T).</p

Oligonucleotide components used in the single well triplex assay for ABCB1 rs2032582.

<p>Multiplexed PlexPrimers are designed in i) Loop, ii) Planar and iii) Target Loop confirmations to reduce competition and produce amplicons that can then be easily distinguished by PlexZymes due to their substantial sequence difference. Only the partzyme target sensor arms of each PlexZyme are illustrated here, showing partzyme A sensor arms (red boxes) which are specific for each variant allele T, A and G (iv, v, vi respectively), and partzyme B sensor arms (blue boxes) which are common to all variant alleles. The additional single mismatch in the primer 3T region is in purple. The target sequence for each variant allele is in black, the complementary regions of the primer sequence is in green, the variant alleles are in red and the INS are in deep red, grey and pink.</p

DNA templates & comparator assays for analysis of ADME SNPs.

<p>DNA templates & comparator assays for analysis of ADME SNPs.</p

Oligonucleotides for analysis of a tri-allelic ADME SNP (dsSNP ID rs2032582) in a single multiplex reaction.

<p>Oligonucleotides for analysis of a tri-allelic ADME SNP (dsSNP ID rs2032582) in a single multiplex reaction.</p