Publications by authors named "Avinash Kaur"

13 Publications

  • Page 1 of 1

Bioluminescent Protein-Inhibitor Pair in the Design of a Molecular Aptamer Beacon Biosensing System.

Anal Chem 2020 06 21;92(11):7393-7398. Epub 2020 May 21.

University of Miami, Leonard M. Miller School of Medicine, Department of Biochemistry and Molecular Biology, Miami, Florida 33136, United States.

Although bioluminescent molecular beacons designed around resonance quenchers have shown higher signal-to-noise ratios and increased sensitivity compared with fluorescent beacon systems, bioluminescence quenching is still comparatively inefficient. A more elegant solution to inefficient quenching can be realized by designing a competitive inhibitor that is structurally very similar to the native substrate, resulting in essentially complete substrate exclusion. In this work, we designed a conjugated anti-interferon-γ (IFN-γ) molecular aptamer beacon (MAB) attached to a bioluminescent protein, luciferase (GLuc), and an inhibitor molecule with a similar structure to the native substrate coelenterazine. To prove that a MAB can be more sensitive and have a better signal-to-noise ratio, a bioluminescence-based assay was developed against IFN-γ and provided an optimized, physiologically relevant detection limit of 1.0 nM. We believe that this inhibitor approach may provide a simple alternative strategy to standard resonance quenching in the development of high-performance molecular beacon-based biosensing systems.
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http://dx.doi.org/10.1021/acs.analchem.0c00518DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7955708PMC
June 2020

Levels of TGF-β1 in peri-miniscrew implant crevicular fluid.

J Oral Biol Craniofac Res 2020 Apr-Jun;10(2):93-98. Epub 2020 Mar 6.

Centre for Biomedical Engineering, Indian Institute of Technology Delhi, New Delhi, 110016, India.

The peri-miniscrew implant crevicular fluid is analogous to gingival crevicular fluid, and its contents reflect the state of inflammation and health during the life of the miniscrews in the mouth. The stability of MSI is fundamental to its role as an anchorage. This study aimed to evaluate transforming growth factor-beta one (TGF-β1) of the peri-miniscrew implant crevicular fluid (PMICF), on implant insertion, pre- and post-loading of MSIs to find a clue to their role in the stability of MSI. Fifty-two MSIs sites were placed in the mouths of 13 patients aged 12-26 years undergoing orthodontic treatment. PMICF was collected using micro-pipettes at T1 (day 0, 1 h after MSI implantation), T2 (day 1), T3/baseline (day 21, preloading of MSI), T4 (day 21, 1 h post loading), T5 (day 22, 1 day post loading), T6 (day 43, 3 weeks post loading). The levels of TGF-β1 were estimated by enzyme-linked immunosorbent assay (ELISA). The data were subjected to statistical analysis. Of the 52 MSIs, 20 MSIs failed at T3. In the case of successful MSIs, the TGF-β1 levels were found to monotonously decrease from T1 (~1400 pg/mL) until T3 (~700 pg/mL) and saturate thereafter. In the case of failed MSIs, the levels of TGF-β1 at various time periods were approximately constant and of much lower value than corresponding time periods of successful MSIs. This study highlights the role of TGF- β1 in bone metabolism around miniscrew reflecting the state of inflammation from 1 h post-implantation.
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http://dx.doi.org/10.1016/j.jobcr.2020.03.002DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7082543PMC
March 2020

Simultaneous and high sensitive detection of Salmonella typhi and Salmonella paratyphi a in human clinical blood samples using an affordable and portable device.

Biomed Microdevices 2019 11 9;21(4):95. Epub 2019 Nov 9.

Centre for Biomedical Engineering, Indian Institute of Technology Delhi, New Delhi, 110016, India.

Enteric fever is one of the leading causes of infection and subsequent fatality (greater than 1.8 million) (WHO 2018), especially in the developing countries due to contaminated water and food inter twinned with unhygienic practices. Clinical gold standard technique of culture-based method followed by biochemical tests demand 72+ hours for diagnosis while newly developed techniques (like PCR, RT-PCR, DNA microarray etc.) suffer from high limit of detection or involve high-cost infrastructure or both. In this work, a quick and highly specific method, SMOL was established for simultaneous detection of Salmonella paratyphi A and Salmonella typhi in clinical blood samples. SMOL consists of (i) pre-concentration of S. typhi and S. paratyphi A cells using magnetic nanoparticles followed by (ii) cell lysis and DNA extraction (iii) amplification of select nucleic acids by LAMP technique and (iv) detection of amplified nucleic acids using an affordable portable device (costs less than $70). To identify the viability of target cells at lower concentrations, the samples were processed at two different time periods of t = 0 and t = 4 h. Primers specific for the SPA2539 gene in S. paratyphi A and STY2879 gene in S. typhi were used for LAMP. Within 6 h SMOL was able to detect positive and negative samples from 55 human clinical blood culture samples and detect the viability of the cells. The results were concordant with culture and biochemical tests as well as by qPCR. Statistical power analysis yielded 100%. SMOL results were concordant with culture and biochemical tests as well as by qPCR. The sensitive and affordable system SMOL will be effective for poor resource settings.
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http://dx.doi.org/10.1007/s10544-019-0441-6DOI Listing
November 2019

Rapid Detection Device for in Milk, Juice, Water and Calf Serum.

Indian J Microbiol 2018 Sep 30;58(3):381-392. Epub 2018 Apr 30.

1Device Testing Laboratory, Centre for Biomedical Engineering, Indian Institute of Technology Delhi, New Delhi, 110016 India.

A limit of detection of 200 CFU/mL of spiked in various sample matrices were achieved in 30 min. The sample matrices were raw/unprocessed milk, commercially available milk, juice from packed bottles, fresh juice from carts, potable water, turbid water and calf serum. The complete protocol comprised of three steps: (a) cell lysis (b) nucleic acid amplification and (c) an in situ optical detection. The cell lysis was carried out using a simple heating based protocol, while the loop-mediated isothermal amplification of DNA was carried out by an in-house designed and fabricated system. The developed system consists of an aluminum block fitted with two cartridge heaters along with a thermocouple. The system was coupled to a light source and spectrometer for a simultaneous in situ detection. Primers specific for STY2879 gene were used to amplify the nucleic acid sequence, isolated from cells. The protocol involves 15 min of cell lysis and DNA isolation followed by 15 min for isothermal amplification and simultaneous detection. No cross-reactivity of the primers were observed at 10 CFU/mL of , , , , . In addition, the system was able to detect of 200 CFU/mL in a concoction of 10 CFU/mL of , 10 CFU/mL of , and 10 CFU/mL of hepatocyte-derived cellular carcinoma HUH7 cells. The proposed rapid diagnostic system shows a promising future in the field of food and medical diagnostics.
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http://dx.doi.org/10.1007/s12088-018-0730-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6023822PMC
September 2018

Highly-sensitive detection of Salmonella typhi in clinical blood samples by magnetic nanoparticle-based enrichment and in-situ measurement of isothermal amplification of nucleic acids.

PLoS One 2018 28;13(3):e0194817. Epub 2018 Mar 28.

Centre for Biomedical Engineering, Indian Institute of Technology Delhi, New Delhi, India.

Enteric fever continues to be a major cause of mortality and morbidity globally, particularly in poor resource settings. Lack of rapid diagnostic assays is a major driving factor for the empirical treatment of enteric fever. In this work, a rapid and sensitive method 'Miod' 'has been developed. Miod includes a magnetic nanoparticle-based enrichment of target bacterial cells, followed by cell lysis and loop-mediated isothermal amplification (LAMP) of nucleic acids for signal augmentation along with concurrent measurement of signal via an in-situ optical detection system. To identify positive/negative enteric fever infections in clinical blood samples, the samples were processed using Miod at time = 0 hours and time = 4 hours post-incubation in blood culture media. Primers specific for the STY2879 gene were used to amplify the nucleic acids isolated from S. typhi cells. A limit of detection of 5 CFU/mL was achieved. No cross-reactivity of the primers were observed against 106 CFU/mL of common pathogenic bacterial species found in blood such as E. coli, P. aeruginosa, S. aureus, A. baumanni, E. faecalis, S. Paratyphi A and K. pneumonia. Miod was tested on 28 human clinical blood samples. The detection of both pre-and post-four-hours incubation confirmed the presence of viable S. typhi cells and allowed clinical correlation of infection. The positive and negative samples were successfully detected in less than 6 hours with 100% sensitivity and specificity.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0194817PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5874042PMC
July 2018

Identification of p38 MAPK as a novel therapeutic target for Friedreich's ataxia.

Sci Rep 2018 03 22;8(1):5007. Epub 2018 Mar 22.

Department of Pathology and Laboratory Medicine, Children's Hospital Philadelphia, Philadelphia, PA, USA.

Friedreich ataxia (FRDA) is an autosomal recessive neuro- and cardio-degenerative disorder caused by decreased expression of frataxin, a protein that localizes to mitochondria and is critical for iron-sulfur-cluster (ISC) assembly. There are no proven effective treatments for FRDA. We previously screened a random shRNA library and identified a synthetic shRNA (gFA11) that reverses the growth defect of FRDA cells in culture. We now report that gFA11 decreases cytokine secretion in primary FRDA fibroblasts and reverts other changes associated with cell senescence. The gene-expression profile induced by gFA11 is remarkably similar to the gene-expression profile induced by the p38 MAPK inhibitor SB203580. We found that p38 phosphorylation, indicating activation of the p38 pathway, is higher in FRDA cells than in normal control cells, and that siRNA knockdown of frataxin in normal fibroblasts also increases p38 phosphorylation. Treatment of FRDA cells with p38 inhibitors recapitulates the reversal of the slow-growth phenotype induced by clone gFA11. These data highlight the involvement of the p38 MAPK pathway in the pathogenesis of FRDA and the potential use of p38 inhibitors as a treatment for FRDA.
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http://dx.doi.org/10.1038/s41598-018-23168-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5864720PMC
March 2018

A review of biomarkers in peri-miniscrew implant crevicular fluid (PMICF).

Prog Orthod 2017 Nov 27;18(1):42. Epub 2017 Nov 27.

Centre for Biomedical Engineering, Indian Institute of Technology Delhi, New Delhi, India.

Background: The temporary anchorage devices (TADs) which include miniscrew implants (MSIs) have evolved as useful armamentarium in the management of severe malocclusions and assist in complex tooth movements. Although a multitude of factors is responsible for the primary and secondary stability of miniscrew implants, contemporary research highlights the importance of biological interface of MSI with bone and soft tissue in augmenting the success of implants. The inflammation and remodeling associated with MSI insertion or loading are reflected through biomarkers in peri-miniscrew implant crevicular fluid (PMICF) which is analogous to the gingival crevicular fluid. Analysis of biomarkers in PMICF provides indicators of inflammation at the implant site, osteoclast differentiation and activation, bone resorption activity and bone turnover. The PMICF for assessment of these biomarkers can be collected non-invasively via paper strips, periopaper or micro capillary pipettes and analysed by enzyme-linked immunosorbent assay (ELISA) or immunoassays. The markers and mediators of inflammation have been previously studied in relation to orthodontic tooth movement include interleukins (IL-1β, IL-2, IL-6 and IL-8), growth factors and other proteins like tumour necrosis factor (TNF-α), receptor activator of nuclear factor kappa-B ligand (RANKL), chondroitin sulphate (CS) and osteoprotegerin (OPG). Studies have indicated that successful and failed MSIs have different concentrations of biomarkers in PMICF. However, there is a lack of comprehensive information on this aspect of MSIs. Therefore, a detailed review was conducted on the subject.

Results: A literature search revealed six relevant studies: two on IL-1β; one on IL-2, IL-6 and IL-8; one on TNF-α; one on CS; and one on RANKL/OPG ratio. One study showed an increase in IL-1β levels upon MSI loading, peak in 24 hours (h), followed by a decrease in 21 days to reach baseline in 300 days. A 6.87% decrease in IL-2 levels was seen before loading and a 5.97% increase post-loading. IL-8 showed a 6.31% increase after loading and IL-6 increased by 3.08% before MSI loading and 15.06% after loading. RANKL/OPG ratio increased in loaded compared to unloaded MSIs.

Conclusions: Cytokines (mainly ILs and TNF-α) and RANKL/OPG ratio showed alteration in PMICF levels upon loading of MSIs as direct or indirect anchorage.
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http://dx.doi.org/10.1186/s40510-017-0195-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5702602PMC
November 2017

Phacoemulsification versus small incision cataract surgery in patients with uveitis.

Int J Ophthalmol 2015 18;8(5):965-70. Epub 2015 Oct 18.

Department of Ophthalmology, Rotary Eye Hospital, Maranda, Palampur 176102, India.

Aim: To compare the safety and efficacy of phacoemulsification and small incision cataract surgery (SICS) in patients with uveitic cataract.

Methods: In a prospective, randomized multi-centric study, consecutive patients with uveitic cataract were randomized to receive phacoemulsification or manual SICS by either of two surgeons well versed with both the techniques. A minimum inflammation free period of 3mo (defined as less than 5 cells per high power field in anterior chamber) was a pre-requisite for eligibility for surgery. Superior scleral tunnel incisions were used for both techniques. Improvement in visual acuity post-operatively was the primary outcome measure and the rate of post-operative complications and surgical time were secondary outcome measures, respectively. Means of groups were compared using t-tests. One way analysis of variance (ANOVA) was used when there were more than two groups. Chi-square tests were used for proportions. Kaplan Meyer survival analysis was done and means for survival time was estimated at 95% confidence interval (CI). A P value of <0.05 was considered statistically significant.

Results: One hundred and twenty-six of 139 patients (90.6%) completed the 6-month follow-up. Seven patients were lost in follow up and another six excluded due to either follow-up less than six months (n=1) or inability implant an intraocular lens (IOL) because of insufficient capsular support following posterior capsule rupture (n=5). There was significant improvement in vision after both the procedures (paired t-test; P<0.001). On first postoperative day, uncorrected distance visual acuity (UDVA) was 20/63 or better in 31 (47%) patients in Phaco group and 26 (43.3%) patients in SICS group (P=0.384). The mean surgically induced astigmatism (SIA) was 0.86±0.34 dioptres (D) in the phacoemulsification group and 1.16±0.28 D in SICS group. The difference between the groups was significant (t-test, P=0.002). At 6mo, corrected distance visual acuity (CDVA) was 20/60 or better in 60 (90.9%) patients in Phaco group and 53 (88.3%) in the manual SICS group (P=0.478). The mean surgical time was significantly shorter in the manual SICS group (10.8±2.9 versus 13.2±2.6min) (P<0.001). Oral prednisolone, 1 mg/kg body weight was given 7d prior to surgery, continued post-operatively and tapered according to the inflammatory response over 4-6wk in patients with previously documented macular edema, recurrent uveitis, chronic anterior uveitis and intermediate uveitis. Rate of complications like macular edema (Chi-square, P=0.459), persistent uveitis (Chi-square, P=0.289) and posterior capsule opacification (Chi-square, P=0.474) were comparable between both the groups.

Conclusion: Manual SICS and phacoemulsification do not differ significantly in complication rates and final CDVA outcomes. However, manual SICS is significantly faster. It may be the preferred technique in settings where surgical volume is high and access to phacoemulsification is limited, such as in eye camps. It may also be the appropriate technique for uveitic cataract under such circumstances.
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http://dx.doi.org/10.3980/j.issn.2222-3959.2015.05.20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4630994PMC
November 2015

Phenotypic Screening for Friedreich Ataxia Using Random shRNA Selection.

J Biomol Screen 2015 Oct 18;20(9):1084-90. Epub 2015 Aug 18.

Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, PA, USA The Penn Medicine/CHOP Center of Excellence for Friedreich's Ataxia Research, Philadelphia, PA, USA

Friedreich ataxia (FRDA) is an autosomal recessive neuro- and cardio-degenerative disorder for which there are no proven effective treatments. FRDA is caused by decreased expression and/or function of the protein frataxin. Frataxin chaperones iron in the mitochondrial matrix and regulates the iron-sulfur cluster (ISC) assembly complex. ISCs are prosthetic groups critical for the function of the Krebs cycle and the mitochondrial electron transport chain. Decreased expression of frataxin is associated with decreased ISC assembly, mitochondrial iron accumulation, and increased oxidative stress, all of which contribute to mitochondrial dysfunction. In media with beta-hydroxybutyrate (BHB) as carbon source, primary FRDA fibroblasts grow poorly and/or lose viability over several days. We screened a random, short-hairpin-RNA (shRNA)-expressing library in primary FRDA fibroblasts and identified two shRNAs that reverse the growth/viability defect in BHB media. One of these two clones increases frataxin expression in primary FRDA fibroblasts, either as a vector-expressed shRNA or as a transfected short-interfering RNA (siRNA).
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http://dx.doi.org/10.1177/1087057115600433DOI Listing
October 2015

A randomized controlled trial of peeling and aspiration of Elschnig pearls and neodymium: yttrium-aluminium-garnet laser capsulotomy.

Int J Ophthalmol 2015 18;8(3):590-6. Epub 2015 Jun 18.

Department of Ophthalmology, Santosh, Medical College and Hospital, Ghaziabad 201301, India.

Aim: To compare surgical peeling and aspiration and neodymium yttrium garnet laser capsulotomy for pearl form of posterior capsule opacification (PCO).

Methods: A prospective, randomized, double blind, study was done at Rotary Eye Hospital, Maranda, Palampur, India, Santosh Medical College Hospital, Ghaziabad, India and Laser Eye Clinic, Noida India. Consecutive patients with pearl form of PCO following surgery, phacoemulsification, manual small incision cataract surgery and conventional extracapsular cataract extraction (ECCE) for age related cataract, were randomized to have peeling and aspiration or neodymium yttrium garnet laser capsulotomy. Corrected distance visual acuity (CDVA), intra-operative and post-operative complications were compared.

Results: A total of 634 patients participated in the study, and 314 (49.5%) patients were randomized to surgical peeling and aspiration group and 320 (50.5%) to the Nd:YAG laser group. The mean pre-procedural logMAR CDVA in peeling and neodymium: yttrium-aluminium-garnet (Nd:YAG) laser group was 0.80±0.25 and 0.86±0.22, respectively. The mean final CDVA in peeling group (0.22±0.23) was comparable to Nd:YAG group (0.24±0.28; t test, P=0.240). There was a significant improvement in vision after both the procedures (P<0.001). A slightly higher percentage of patients in Nd:YAG laser group (283/88.3%) than in peeling group (262/83.4%) had a CDVA of 0.5 (20/63) or better at 9mo (P<0.001). On the contrary, patients having CDVA worse than 1.00 (20/200) was also significantly higher in Nd:YAG laser group as compared to peeling group (25/7.7% vs 15/4.7%, respectively). On application of ANCOVA, there was less than 0.001% risk that PCO thickness and total laser energy had no effect on rate of complications in Nd:YAG laser group and less than 0.001 % risk that PCO thickness had no effect on complications in peeling group respectively. Sum of square analysis suggests that in the Nd:YAG laser group, thick PCO had a stronger impact on complications (Fischer test probability, Pr<0.0001) than thin PCO and total laser energy (Fischer test probability, Pr<0.002), respectively; similarly, in peeling group, thick PCO and preoperative vision had a stronger effect on complications than thin PCO, respectively (Fischer test probability, Pr<0.001).The rate of complications like uveitis (P=0.527) and cystoid macular edema (P=0.068), did not differ significantly between both the groups. However, intraocular pressure spikes (P=0.046) and retinal detachment (P<0.001) were significantly higher in Nd:YAG laser group as compared to peeling group. Retinal detachment was more common in patients having degenerative myopia (7/87.5%, P<0.001). Recurrence of pearls was the most common cause of reduction of vision in the peeling group (24/7.6%, P<0.001).

Conclusion: There is no alternative to Nd:YAG laser capsulotomy for fibrous subtype of PCO. For pearl form of PCO, both techniques are comparable with regard to visual outcomes. Nd:YAG laser capsulotomy has a higher incidence of IOP spikes and retinal detachment whereas recurrence of pearls may occur after successful peeling and aspiration. When posterior capsulotomy is needed in patients with retinal degenerations, retinopathies and pre-existing retinal breaks, the clinician should be cautious about increased risks of possible complications of Nd:YAG laser capsulotomy.
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http://dx.doi.org/10.3980/j.issn.2222-3959.2015.03.28DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4458669PMC
June 2015

Oral omega-3 fatty acids treatment in computer vision syndrome related dry eye.

Cont Lens Anterior Eye 2015 Jun 16;38(3):206-10. Epub 2015 Feb 16.

Department of Microbiology, Narayan Medical College, Sasaram, India.

Purpose: To assess the efficacy of dietary consumption of omega-3 fatty acids (O3FAs) on dry eye symptoms, Schirmer test, tear film break up time (TBUT) and conjunctival impression cytology (CIC) in patients with computer vision syndrome.

Setting And Design: Interventional, randomized, double blind, multi-centric study.

Methods: Four hundred and seventy eight symptomatic patients using computers for more than 3h per day for minimum 1 year were randomized into two groups: 220 patients received two capsules of omega-3 fatty acids each containing 180mg eicosapentaenoic acid (EPA) and 120mg docosahexaenoic acid (DHA) daily (O3FA group) and 236 patients received two capsules of a placebo containing olive oil daily for 3 months (placebo group). The primary outcome measure was improvement in dry eye symptoms and secondary outcome measures were improvement in Nelson grade and an increase in Schirmer and TBUT scores at 3 months.

Results: In the placebo group, before dietary intervention, the mean symptom score, Schirmer, TBUT and CIC scores were 7.5±2, 19.9±4.7mm, 11.5±2s and 1±0.9 respectively, and 3 months later were 6.8±2.2, 20.5±4.7mm, 12±2.2s and 0.9±0.9 respectively. In the O3FA group, these values were 8.0±2.6, 20.1±4.2mm, 11.7±1.6s and 1.2±0.8 before dietary intervention and 3.9±2.2, 21.4±4mm, 15±1.7s, 0.5±0.6 after 3 months of intervention, respectively.

Conclusion: This study demonstrates the beneficial effect of orally administered O3FAs in alleviating dry eye symptoms, decreasing tear evaporation rate and improving Nelson grade in patients suffering from computer vision syndrome related dry eye.
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http://dx.doi.org/10.1016/j.clae.2015.01.007DOI Listing
June 2015

The diagnostic value and accuracy of conjunctival impression cytology, dry eye symptomatology, and routine tear function tests in computer users.

J Lab Physicians 2014 Jul;6(2):102-8

Department of Health, Medical officer, Uttar Pradesh, India.

Aims And Objectives: To compare the diagnostic value and accuracy of dry eye scoring system (DESS), conjunctival impression cytology (CIC), tear film breakup time (TBUT), and Schirmer's test in computer users.

Methods: A case-control study was done at two referral eye centers. Eyes of 344 computer users were compared to 371 eyes of age and sex matched controls. Dry eye questionnaire (DESS) was administered to both groups and they further underwent measurement of TBUT, Schirmer's, and CIC. Correlation analysis was performed between DESS, CIC, TBUT, and Schirmer's test scores. A Pearson's coefficient of the linear expression (R (2)) of 0.5 or more was statistically significant.

Results: The mean age in cases (26.05 ± 4.06 years) was comparable to controls (25.67 ± 3.65 years) (P = 0.465). The mean symptom score in computer users was significantly higher as compared to controls (P < 0.001). Mean TBUT, Schirmer's test values, and goblet cell density were significantly reduced in computer users (P < 0.001). TBUT, Schirmer's, and CIC were abnormal in 48.5%, 29.1%, and 38.4% symptomatic computer users respectively as compared to 8%, 6.7%, and 7.3% symptomatic controls respectively. On correlation analysis, there was a significant (inverse) association of dry eye symptoms (DESS) with TBUT and CIC scores (R (2) > 0.5), in contrast to Schirmer's scores (R(2) < 0.5). Duration of computer usage had a significant effect on dry eye symptoms severity, TBUT, and CIC scores as compared to Schirmer's test.

Conclusion: DESS should be used in combination with TBUT and CIC for dry eye evaluation in computer users.
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http://dx.doi.org/10.4103/0974-2727.141507DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4196356PMC
July 2014