Objective: To determine the genetic diversity, natural selection and mutations in Plasmodium (P.) knowlesi drug resistant molecular markers Kelch 13 and dhps gene in clinical samples of Malaysia. Methods: P. knowlesi full-length gene sequences Kelch 13 gene (PkK13) from 40 samples and dhps gene from 30 samples originating from Malaysian Borneo were retrieved from public databases. Genetic diversity, natural selection, and phylogenetic analysis of gene sequences were analysed using DNAsp v5.10 and MEGA v5.2. Results: Seventy-two single nucleotide polymorphic sites (SNPs) across the full-length PkK13 gene (63 synonymous substitutions and 9 non-synonymous substitutions) with nucleotide diversity of π~0.005 was observed. Analysis of the full-length Pkdhps gene revealed 73 SNPs and π~0.006 (44 synonymous substitutions and 29 non-synonymous substitutions). A high number of haplotypes (PkK13; H=37 and Pkdhps; H=29) with haplotype diversity of Hd ~0.99 were found in both genes, indicating population expansion. Nine mutant alleles were identified in PkK13 amino acid alignment of which, 7 (Asp 3 Glu, Lys 50 Gln, Lys 53 Glu, Ser 123 Thr, Ser 127 Pro, Ser 149 Thr and Ala 169 Thr) were within the Plasmodium specific domain, 2 (VaI 372 Ile and Lys 424 Asn) were in the BTB/POZ domain and no mutation was observed within the kelch propeller domain. The 29 non-synonymous mutations in the Pkdhps gene were novel and only presented in exon 1 and 2. Conclusions: Monitoring the mutations from clinical samples collected from all states of Malaysia along with clinical efficacy studies will be necessary to determine the drug resistance in P. knowlesi.

BACKGROUND: Vitiligo is a depigmenting skin disorder in which genetic factors play an important role. OBJECTIVE: To examine the association of CYP2C9 *1/*2/*3 gene polymorphism with vitiligo. METHODS: In this case controlled study, 95 Saudi patients with vitiligo (50 men and 45 women), with a mean age of 27.3 years, were analyzed. Patients were compared to 86 healthy controls from the same locality (76 men and 10 women), with a mean age of 20.1 years. In all participants, DNA was extracted and processed for characterization of 2C9 *1/*2/*3 gene variants using real time-polymerase chain reaction. RESULTS: Vitiligo patients have a significantly higher CYP2C9 *3 allele carriage rate compared to controls (32.7% versus 4.7%, p=0.00, odds ratio=9.9, 95% confidence interval=3.3~29.6). On the other hand, frequencies of CYP2C9 *2 genotypes and alleles did not show any significant difference between vitiligo cases and controls. When the frequencies of CYP2C9 genotypes were compared among subgroups of age, gender, family history, and disease patterns, the cases with positive consanguinity had significantly higher frequencies of homozygous genotypes than others (p=0.029). CONCLUSION: CYP2C9 *3 allele carriage is probably associated with vitiligo susceptibility.
Alleles ; Case-Control Studies ; Consanguinity ; DNA ; Genotype ; Hand ; Humans ; Male ; Polymorphism, Genetic ; Skin ; Vitiligo*

INTRODUCTIONType 2 diabetes (T2D) candidate gene: potassium voltage-gated channel, KQT-like subfamily, member 1 (KCNQ1) was suggested by conducting a genome wide association study (GWAS) in Japanese population. Association studies have been replicated among East Asian populations; however, the association between this gene and T2D in Southeast Asian populations still needs to be studied. This study aimed to investigate the association of KCNQ1 common variants with type 2 diabetes in Malaysian Malay subjects.

MATERIALS AND METHODSThe KCNQ1 single nucleotide polymorphisms (SNPs): rs2237892, rs2283228, and rs2237895 were genotyped in 234 T2D and 177 normal Malay subjects.

RESULTSThe risk allele of the rs2283228 (A) was strongly associated with T2D (OR = 1.7, P = 0.0006) while the rs2237892 (C) was moderately associated with T2D (OR = 1.45, P = 0.017). The recessive genetic models showed that rs2283228 was strongly associated with T2D (OR = 2.35, P = 0.00005) whereas rs2237892 showed a moderate association with T2D (OR = 1.69, P = 0.01). The haplotype block (TCA), which contained the protective allele, correlated with a protection from T2D (OR = 0.5, P = 0.003). Furthermore, the diplotype (CAA-TCA) that contained the protective haplotype was protected against T2D (OR = 0.46, P = 0.006).

CONCLUSIONThe KCNQ1 SNPs, haplotypes and diplotypes are associated with T2D in the Malaysian Malay subjects.

Adult ; Diabetes Mellitus, Type 2 ; ethnology ; genetics ; Female ; Genetics, Population ; Haplotypes ; genetics ; Humans ; KCNQ1 Potassium Channel ; genetics ; Malaysia ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; genetics ; Sequence Analysis, DNA

AIM: To evaluate the use of amniotic membrane transplantation(AMT)in symblepharon.

METHODS: This non-comparative interventional case study was conducted from January 2013 to December 2015 and included a consecutive series of 14 eyes of 12 patients. Patients were selected for permanent AMT. The amniotic patches were grafted for the treatment of symblepharon. Cryo-preserved or freeze-dried amniotic membrane(AM)was used. Regarded to the 14 eyes(12 patients), their age was ranged from 26-62y, with the mean age of 43.38±11.25, 10 eyes of 8 patients(4 males/4 females)were presented with symblepharon secondary to previous pterygium surgery, and 4 eyes of 4 patients(1 male/3 females)were presented with symblepharon secondary to previous strabismus surgery, at least 6mo after the last surgery. The outcome of success was defined as restoration of a stable-depth fornix and being free of scar or inflammation, and no motility restriction during the follow up of 6mo.

RESULTS: The mean follow-up period was 7±4.2mo(range 6-9mo). In all 14 eyes, complete epithelialization of AM was observed 3wk after surgery, resulting in a non-inflamed appearance of the surgical site. Eight eyes out of total 14 eyes showed successful fornix reconstruction with success rate(57%), the fornix was deep, and no recurrence was observed. Four eyes(29%)showed partial success with moderate depth of the fornix and moderate scar. Two eyes(14%)showed failure of reconstruction of the fornix with complete fornix obliteration. The visual acuity improved after surgery in 7 eyes while remained stable in 7 eyes. Post-operative complications from the AMT was very limited as severe conjunctival reaction and motility restriction was occurred only in one eye out of 14 eyes(7%)and pyogenic granuloma occurred in 2 eyes out of 14 eyes(14%)in the first 3mo after surgery and was managed with surgical excision, with local corticosteroid injection.

CONCLUSION: AMT alone is a safe and effective method for symblepharon. Considering the potential adverse effects associated with limbal excision, also, AMT is an effective method of fornix reconstruction for the repair of symblepharon in a variety of ocular surface disorders.

The complement cascade is a central component of innate immunity which plays a critical role in brain inflammation. Complement C3a receptor (C3aR) is a key mediator of post-ischemic cerebral injury, and pharmacological antagonism of the C3a receptor is neuroprotective in stroke. Cerebral ischemia injures brain endothelial cells, causing blood brain barrier (BBB) disruption which further exacerbates ischemic neuronal injury. In this study, we used an in vitro model of ischemia (oxygen glucose deprivation; OGD) to investigate the protective effect of a C3aR antagonist (C3aRA, SB290157) on brain endothelial cells (bEnd.3). Following 24 hours of reperfusion, OGD-induced cell death was assessed by TUNEL and Caspase-3 staining. Western blot and immunocytochemistry were utilized to demonstrate that OGD upregulates inflammatory, oxidative stress and antioxidant markers (ICAM-1, Cox-2, Nox-2 and MnSOD) in endothelial cells and that C3aRA treatment significantly attenuate these markers. We also found that C3aRA administration restored the expression level of the tight junction protein occludin in endothelial cells following OGD. Interestingly, OGD/reperfusion injury increased the phosphorylation of ERK1/2 and C3aR inhibition significantly reduced the activation of ERK suggesting that endothelial C3aR may act via ERK signaling. Furthermore, exogenous C3a administration stimulates these same inflammatory mechanisms both with and without OGD, and C3aRA suppresses these C3a-mediated responses, supporting an antagonist role for C3aRA. Based on these results, we conclude that C3aRA administration attenuates inflammation, oxidative stress, ERK activation, and protects brain endothelial cells following experimental brain ischemia.
Blood-Brain Barrier ; Blotting, Western ; Brain Ischemia ; Brain ; Caspase 3 ; Cell Death ; Complement C3a ; Complement System Proteins ; Encephalitis ; Endothelial Cells ; Glucose ; Immunity, Innate ; Immunohistochemistry ; In Situ Nick-End Labeling ; In Vitro Techniques ; Inflammation ; Ischemia ; Neurons ; Occludin ; Oxidative Stress ; Phosphorylation ; Reperfusion ; Stroke ; Tight Junctions

Objective To summarize the precise association between pulmonary tuberculosis (PTB) and P2x7 A1513C gene polymorphism. Methods PubMed and Google Scholar web-databases were searched for the studies reporting the association of P2x7 A1513C polymorphism and PTB risk. A meta-analysis was performed for the selected case–control studies and pooled odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated for all the genetic models. Results Eleven studies comprising 2 678 controls and 2 113 PTB cases were included in this meta-analysis. We observed overall no significant risk in all the five genetic models. When stratified population by the ethnicity, Caucasian population failed to show any risk of PTB in all the genetics models. In Asian ethnicity, variant allele (C vs. A: P = 0.001; OR = 1.375, 95% CI = 1.159–1.632) and heterozygous genotype (AC vs. AA: P = 0.001; OR = 1.570, 95% CI = 1.269–1.944) demonstrated significant increased risk of PTB. Likewise, recessive genetic model (CC + AC vs. AA: P = 0.001; OR = 1.540, 95% CI = 1.255–1.890) also demonstrated increased risk of PTB in Asians. Conclusions Our meta-analysis did not suggest the association of P2x7 A1513C polymorphism with PTB risk in overall or separately in Caucasian population. However, it plays a significant risk factor for predisposing PTB in Asians. Future larger sample and expression studies are needed to validate this association.