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Identification of potential serum metabolic biomarkers for patient with keratoconus using untargeted metabolomics approach

Daphne Teh AL, Jayapalan JJ, Loke MF, Wan Abdul Kadir AJ, Subrayan V

Exp Eye Res, 2021 10;211:108734.
PMID: 34428458 DOI: 10.1016/j.exer.2021.108734

This study aimed to investigate the metabolite differences between patients with keratoconus and control subjects and identify potential serum biomarkers for keratoconus using a non-targeted metabolomics approach. Venous blood samples were obtained from patients with keratoconus (n = 20) as well as from age-, gender- and race-matched control subjects (n = 20). Metabolites extracted from serum were separated and analyzed by liquid chromatography/quadrupole time-of-flight mass spectrometer. Processing of raw data and analysis of the data files was performed using Agilent Mass Hunter Qualitative software. The identified metabolites were subjected to a principal component and hierarchical cluster analysis. Appropriate statistical tests were used to analyze the metabolomic profiling data. Together, the analysis revealed that the dehydroepiandrosterone sulfate from the steroidal hormone synthesis pathway was significantly upregulated in patients with keratoconus (p
The Effect of Irradiated Riboflavin in Human Tenon's Fibroblast - A Study on Cellular Viability

See WYN, Ismail F, Sheikh Abdul Kadir SH, Subrayan V

Curr Eye Res, 2021 Dec 29.
PMID: 34963422 DOI: 10.1080/02713683.2021.2011326

PURPOSE/ AIM: The main purpose of this work is to study the cellular viability effect of irradiated riboflavin in cultured human tenon fibroblasts.
MATERIALS AND METHODS: The tenon tissue was harvested from a patient undergoing strabismus surgery. The human tenon fibroblast cell culture and isolation were performed according to the standard laboratory cell culturing protocol. The cells were divided into three groups: control, treatment with irradiated and non-irradiated riboflavin. There were five different concentrations (0.00156%, 0.003125%, 0.00625%, 0.0125%, 0.025%) in each group of riboflavin. The fibroblasts were treated with riboflavin and the cellular viability was assessed at 24-hour and 48-hour post treatment with MTT 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyl tetrazolium bromide colorimetric assay. The absorbance values were analysed using Magellan microplate reader data analysis. A triplicate of readings was taken. The data were presented as mean ± standard deviation of the triplicates. Statistical analysis was performed with Statistical Package for Social Sciences (SPSS) analysis version 23.
RESULTS: Irradiated riboflavin caused a concentration-dependent cell death in human tenon fibroblast cell culture (p
I think I have double vision? Or not? Internuclear Ophthalmoplegia following right lacunar infarct

Lee HN, Subrayan V

Med J Malaysia, 2021 Nov;76(6):950-952.
PMID: 34806694

Internuclear Ophthalmoplegia (INO) is an inability of the ipsilateral adduction with a contralateral horizontal abducting saccade on attempted gaze to the contra-lesion side. Injury to the medial longitudinal fasciculus (MLF) will obstruct the signalling pathway between the ipsilateral abducens nucleus and the contralateral medial rectus muscle. Infarction accounts for 38% of INO cases with mostly being unilateral (87%), followed by demyelination (34%), which mostly being bilateral (73%). Lacunar infarct is the most common ischemic stroke. INO can be easily missed due to its subtle presentation with no complaints from the patients. A full cranial nerves assessment, includes the extraocular muscles movement, is important. Ischemic and demyelinating INO typically recover. We present here of a case of INO following right lacunar infarct in a 72-year-old Malay woman. She had hypertensive crisis due to missed medications. Her blood pressure was well controlled throughout the hospital admission and finally she was discharged home with continuation of care at her primary facility.
Diabetes and pachymetry changes in pregnancy

Santiagu F, Bakhtiari A, Iqbal T, Khaliddin N, Lansingh VC, Subrayan V

Int Ophthalmol, 2018 Oct;38(5):2069-2076.
PMID: 28879527 DOI: 10.1007/s10792-017-0701-5

BACKGROUND: Purpose of this study is to evaluate changes in the central corneal thickness (CCT) in patients during the third trimester and postpartum phases of normal pregnancy, pregestational diabetes mellitus (DM), and gestational diabetes mellitus (GDM).
METHODS: This was a prospective study. Patients that fulfilled the inclusion criteria were recruited from the obstetric clinic. They were grouped into normal pregnancy, pregestational DM, and GDM. Ophthalmic assessment and haematological investigations were done during the third trimester (after 28 weeks of gestation) and in the postpartum phase (6 weeks postpartum).
RESULTS: A total of 192 pregnant patients were recruited for this study. Out of the 192 patients, only 143 of them came back for their follow-up 6 weeks postpartum. A total of 70 (36.5%) normal pregnancy patients, 51 (26.6%) DM patients, and 71 (36.9%) GDM patients were included in this prospective study. Our study showed that the CCT decreased postpartum in all three groups. Patients in all three groups did not have significantly different CCT during the third trimester of pregnancy and postpartum phase. However, patients who had thicker CCT irrespective of the grouping during the third trimester also had a thicker CCT post-delivery (p value <0.001). However, these changes did not appear to affect refractive error and visual acuity.
CONCLUSIONS: Diabetes mellitus during pregnancy did not appear to influence the CCT.
Study site: Obstetrics clinic, University Malaya Medical Center (UMMC), Kuala Lumpur, Malaysia
Fulltext Association between Ocular Pseudoexfoliation and Sensorineural Hearing Loss

Singham NV, Zahari M, Peyman M, Prepageran N, Subrayan V

J Ophthalmol, 2014;2014:825936.
PMID: 24864196 DOI: 10.1155/2014/825936

Background. Our study aimed to investigate an association between ocular pseudoexfoliation (PXF) and sensorineural hearing loss (SNHL) and to compare them with age and sex matched controls without pseudoexfoliation. Method. This was a case-control study of 123 patients which included 68 cases with PXF (at least one eye) and 55 controls without pseudoexfoliation. Pure-tone audiometry (PTA) was done for these patients at sound frequencies taken as important for speech comprehension, that is, 250 Hertz (Hz), 500 Hz, 1000 Hz, and 2000 Hz. Results. There were 41 patients with pseudoexfoliation syndrome (PXE) and 27 with pseudoexfoliative glaucoma (PXEG). The majority of patients with hearing loss (60%; n = 51) were PXF patients and the remaining 40% (n = 34) were controls. Below average hearing thresholds were significantly higher in the pseudoexfoliation group compared to the control group (P = 0.01; odds ratio (OR), 3.00; 95% confidence interval (CI), 1.25-7.19). However, there was no significant difference in the mean hearing threshold levels between the three groups (PXE, PXEG, and controls) in either ear (ANOVA, right ear: P = 0.46 and left ear P = 0.36). Conclusion. Our study found an association between PXF and SNHL, confirming that PXF can involve organs in the body other than the eye.
Fulltext Retinal manifestations of patients with human immunodeficiency virus, a multiethnics study in Malaysia

Loo AV, Sujaya S, Peyman M, Florence S, Subrayan V

Int J Ophthalmol, 2011;4(6):641-3.
PMID: 22553736 DOI: 10.3980/j.issn.2222-3959.2011.06.13

To investigate the fundus findings of patients infected with human immunodeficiency virus (HIV) in correlation to Highly Active Antiretroviral Therapy (HAART) and CD4 count.
Plasma, aqueous and vitreous homocysteine levels in proliferative diabetic retinopathy

Lim CP, Loo AV, Khaw KW, Sthaneshwar P, Khang TF, Hassan M, et al.

Br J Ophthalmol, 2012 May;96(5):704-7.
PMID: 22353698 DOI: 10.1136/bjophthalmol-2011-301044

To compare homocysteine (Hcy) concentration in the blood plasma, vitreous and aqueous of eyes with proliferative diabetic retinopathy (PDR) against control, and to investigate associations between Hcy concentration in blood plasma with that of aqueous and vitreous in these two groups.
Dosage-dependent reduction of macular pigment optical density in female breast cancer patients receiving tamoxifen adjuvant therapy

Lim IL, Loo AVP, Subrayan V, Khang TF, See MH, Alip A, et al.

Breast, 2018 Jun;39:117-122.
PMID: 29660599 DOI: 10.1016/j.breast.2018.04.003

It is now increasingly common for breast cancer patients to receive adjuvant tamoxifen therapy for a period of up to 10 years. As survival rate increases, managing tamoxifen ocular toxicities is important for patients' quality of life. Macular pigments in photoreceptor cells protect against free radical damage, which can cause macular degeneration. By reducing macular pigment concentration, tamoxifen may increase the risk of macular degeneration. Here, we compared macular pigment optical density (MPOD) and central macular thickness between breast cancer patients on tamoxifen adjuvant therapy (n = 70), and a control group (n = 72). Multiple regression analysis indicated that MPOD decreases with increasing tamoxifen dosage, up to a threshold of about 20 g, after which MPOD plateaus out. Mean MPOD in the treatment group (mean = 0.40) was significantly lower (p-value = 0.02) compared to the control group (mean = 0.47) for the left eye, and for the right eye (treatment mean = 0.39; control mean = 0.48; p-value = 0.009). No significant difference in mean central macular thickness was found between the treatment and the control group (p-values > 0.4). In the control group, MPOD and central macular thickness showed significant correlation (r∼0.30; p-values 
Changes in anterior chamber depth and intraocular pressure after phacoemulsification in eyes with occludable angles

Shin HC, Subrayan V, Tajunisah I

J Cataract Refract Surg, 2010 Aug;36(8):1289-95.
PMID: 20656150 DOI: 10.1016/j.jcrs.2010.02.024

PURPOSE: To evaluate changes in anterior chamber depth (ACD) and intraocular pressure (IOP) after phacoemulsification in eyes with occludable angles and compare the results with those in eyes with normal open angles.
SETTING: Eye Clinic, Ipoh General Hospital, Ipoh, Perak, Malaysia.
METHODS: Patients with nonglaucomatous eyes with open angles or with occludable angles were recruited. Ocular biometric measurements (ACD, axial length [AL], lens thickness and position) and IOP (tonometry) were performed preoperatively and 1 day and 1, 4, 9, and 12 weeks postoperatively.
RESULTS: The open-angle group and occludable-angle group each comprised 35 patients. The occludable-angle group had a shallower ACD, shorter AL, thicker and relatively anterior lens position, and higher IOP preoperatively; postoperatively, there was a significant increase in ACD and a significant reduction in IOP (P
Fulltext Endoscopic orbital decompression for optic neuropathy in thyroid ophthalmopathy

Tang IP, Prepageran N, Subrayan V, Tajunisah I

Med J Malaysia, 2008 Oct;63(4):337-8.
PMID: 19385499 MyJurnal

Optic neuropathy, an uncommon manifestation occurs in 5% of patients with Graves' ophthalmopathy. Its outcome is good if early and proper treatment given. We reported a 49 years old male patient of Graves' disease with bilateral optic neuropathy. He underwent bilateral transnasal endoscopic orbital decompression with marked improvement of visual acuity bilaterally. The preoperative visual acuity was 6/36 in his right eye and 6/60 in his left eye. Two days after operation, his visual acuity was 6/12 in his right eye and 6/24 in his left eye. Transnasal endoscopic orbital decompression is a choice of treatment in optic neuropathy in thyroid ophthalmopathy after trial of corticosteroids.
Combined optic neuropathy and central retinal artery occlusion in presumed ocular tuberculosis without detectable systemic infection

Ooi YL, Tai LY, Subrayan V, Tajunisah I

Ocul Immunol Inflamm, 2011 Oct;19(5):370-2.
PMID: 21834697 DOI: 10.3109/09273948.2011.594203

To report a rare case of combined optic neuropathy and central retinal artery occlusion in presumed ocular tuberculosis without systemic infection.
Intravitreal bevacizumab for radiation-induced cystoid macular oedema in patients with nasopharyngeal carcinoma: a clinical series

Subrayan V, Khaw KW, Peyman M, Koay AC, Tajunisah I

Ophthalmologica, 2013;229(4):208-11.
PMID: 23548379 DOI: 10.1159/000348630

To evaluate the outcome of intravitreal bevacizumab in the treatment of radiation-induced cystoid macular oedema among patients who underwent external beam radiotherapy for nasopharyngeal carcinoma.
Visual System Homeobox 1 (VSX1) Gene Analysis in Keratoconus: Design of Specific Primers and DNA Amplification Protocols for Accurate Molecular Characterization

Ng JB, Poh RY, Lee KR, Subrayan V, Deva JP, Lau AY, et al.

Clin. Lab., 2016 Sep 01;62(9):1731-1737.
PMID: 28164597 DOI: 10.7754/Clin.Lab.2016.160144

BACKGROUND: Keratoconus is an ocular degeneration characterized by the thinning of corneal stroma that may lead to varying degrees of myopia and visual impairment. Genetic factors have been reported in the pathology of keratoconus where Asians have a higher incidence, earlier onset, and undergo earlier corneal grafts compared to Caucasians. The visual system homeobox 1 (VSX1) gene forms part of a paired-like homeodomain transcription factor which is responsible for ocular development. The gene was marked as a candidate in genetic studies of keratoconus in various populations. Single nucleotide polymorphisms (SNPs) in the VSX1 gene have been reported to be associated with keratoconus. The detection of the SNPs involves DNA amplification of the VSX1 gene followed by genomic sequencing. Thus, the objective of this study aims to establish sensitive and accurate screening protocols for the molecular characterization of VSX1 polymorphisms.
METHODS: Keratoconic (n = 74) and control subjects (n = 96) were recruited based on clinical diagnostic tests and selection criteria. DNA extracted from the blood samples was used to genotype VSX1 polymorphisms. In-house designed primers and optimization of PCR conditions were carried out to amplify exons 1 and 3 of the VSX1 gene. PCR conditions including percentage GC content, melting temperatures, and differences in melting temperatures of primers were evaluated to produce sensitive and specific DNA amplifications.
RESULTS: Genotyping was successfully carried out in 4 exons of the VSX1 gene. Primer annealing temperatures were observed to be crucial in enhancing PCR sensitivity and specificity. Annealing temperatures were carefully evaluated to produce increased specificity, yet not allowing sensitivity to be compromised. In addition, exon 1 of the VSX1 gene was amplified using 2 different sets of primers to produce 2 smaller amplified products with absence of non-specific bands. DNA amplification of exons 1 and 3 consistently showed single band products which were successfully sequenced to yield reproducible data.
CONCLUSIONS: The use of in-house designed primers and optimized PCR conditions allowed sensitive and specific DNA amplifications that produced distinct single bands. The in-house designed primers and DNA amplification protocols established in this study provide an addition to the current repertoire of primers for accurate molecular characterization of VSX1 gene polymorphisms in keratoconus research.
Effects of ranibizumab on TGF-β1 and TGF-β2 production by human Tenon's fibroblasts: An in vitro study

Noh SM, Abdul Kadir SH, Crowston JG, Subrayan V, Vasudevan S

Mol Vis, 2015;21:1191-200.
PMID: 26539031

Inhibiting exaggerated wound healing responses, which are primarily mediated by human Tenon's fibroblast (HTF) migration and proliferation, has become the major determining factor for a successful trabeculectomy. Antivascular endothelial growth factor (anti-VEGF) has showed promising results as a potential antifibrotic candidate for use concurrently in trabeculectomy. Preliminary cohort studies have revealed improved bleb morphology following trabeculectomy augmented with ranibizumab. However, the effects on HTFs remain unclear. This study was conducted to understand the effects of ranibizumab on transforming growth factor (TGF)-β1 and transforming growth factor (TGF)-β2 expression by HTFs.
Fulltext Association of copy number variations in complement factor H-Related genes among age-related macular degenerative subjects

Bakri NM, Ramachandran V, Kee HF, Subrayan V, Isa H, Ngah NF, et al.

Kaohsiung J. Med. Sci., 2017 Dec;33(12):602-608.
PMID: 29132549 DOI: 10.1016/j.kjms.2017.08.003

Age-related macular degeneration (AMD) is the most widely recognised cause of irreversible vision loss and previous studies have suggested that the advancement of wet AMD is influenced by both modifiable and non-modifiable elements. Single nucleotide polymorphism (SNPs) and copy number of variations (CNVs) have been associated with AMD in various populations, however the results are conflicting. Our aim is to determine the CNVs of Complement Factor H-Related genes among Malaysian subjects with wet AMD. 130 patients with wet AMD and 120 healthy controls were included in this research. DNA was extracted from all subjects and CNVs of CFH, CFHR1 and CFHR3 genes; determined using quantitative real-time PCR and were compared between the two groups. A consistent association was observed between CFH gene and wet AMD susceptibility (P
Minimally invasive image-guided removal of retrobulbar intraconal foreign body

Vairavan N, Tajunisah I, Subrayan V, Waran V

Orbit, 2009;28(6):442-3.
PMID: 19929682 DOI: 10.3109/01676830903103365

Surgical approaches are becoming increasingly minimally invasive, without compromising either safety or ease. Penetrating ocular foreign bodies has traditionally been approached either by intraocular or supraorbital access. We successfully attempted a minimally invasive approach to remove a retrobulbar foreign body under computer-assisted image guidance in a 19-year-old man involved in an industrial mishap.

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