Purpose: Fusion transcripts have been reported as biomarkers for several diseases; however, a fusion gene has not been reported in ophthalmic disease. To the best of our knowledge, the discovery of fusion transcript in Behcet’s disease (BD), a noninfectious uveitis, is reported for the first time. We generated complete transcript data for BD subtape and discovered fusion transcripts specific to BD patients. Methods: We sequenced total RNA from peripheral blood mononuclear cells isolated from clinically characterized BD patients and controls and generated BAM files, which served as input data for predicting fusion transcripts. We used Arriba, a computational tool for the detection of fusion transcripts. Arriba is fast and accurate for the identification of gene fusions from RNA sequencing data. It is specifically designed for high-throughput RNA-seq data, leveraging STAR-aligned chimeric BAM files to detect fusion events with high sensitivity. Results: The Arriba tool identified a set of fusion transcripts specific to BD patients. One of the fusion transcripts was characterized and validated, which represents the read-through fusion with a product size of 421 bp consisting of a part of exon 2 of NFATC2 (112bp) and a part of exon 28 of ATP9A (309 bp); these are adjacent genes in chromosome 20. Conclusion: This paper reports the discovery of fusion transcript in uveitis-BD subtype and is also the first report for any ophthalmic disease. Such fusion transcript is absent in controls and other subtypes of uveitis. It may be a possible biomarker for the noninfectious BD patients.

To identify genomic risk factors for diabetic retinopathy (DR), proliferative diabetic retinopathy (PDR) and diabetic macular edema (DME), in South Indians. Phenotyping, including optical coherence tomography (OCT), of South Indians (n = 2538) with type 2 diabetes (T2D) was obtained. Genome-wide association studies (GWAS) were performed for DR, PDR, and DME with covariate adjustment. The results were replicated in two cohorts. T2D polygenic risk scores (PRS) were examined to determine if DR cases were enriched for T2D loci. A novel locus on chromosome 10 [rs11199996 (OR = 4.10, P = 2.88 × 10–8)] was associated with PDR, replicated in other cohorts, and is located near genes involved in retinal light transduction and growth hormone signaling. Several near genome-wide significant loci were identified, including rs76323047 in the glucokinase gene (GCK, P = 3.89 × 10–7), a glucose sensor; this risk variant displays higher frequency in Asian populations. T2D PRS showed consistent associations with DR, particularly PDR, suggesting that DR cases have a higher genetic load for T2D. This first DR GWAS in South Indians identified a significant association between a chromosome 10 variant and PDR. We also found notable associations of T2D PRS with DR.

Abstract Background Widespread use of retinal optical coherence tomography angiography (OCT-A) imaging in population-based studies requires methodological and analytical consensus to ensure reproducible and accurate results. Objective Development of a consensus framework for assessment of OCT-A image quality and working with OCT-A-derived variables in population studies. Methods Criteria for image quality were developed for fovea-centered, 3x3-mm OCT-A images acquired with three commercially available devices. Inter- and intragrader agreements for overall image quality, vessel density (VD) and foveal avascular zone (FAZ) area were evaluated among five graders (% agreement). Intergrader agreement was validated on 6x6-mm OCT-A images. Recommendations for grading and use of OCT-A images in epidemiological studies were developed. Results Mean intergrader agreements for overall Unusable, Usable, and Excellent 3x3-mm OCT-A image quality (52 images) were 82.3%, 70.8%, and 87.1%; for Unusable VD and FAZ area agreements were 80.0% and 81.9%, respectively. Mean intragrader agreements for overall Unusable, Usable and Excellent 3x3-mm OCT-A image quality (27 images) were 91.1%, 82.2%, and 81.9%; agreements for Unusable VD and FAZ area were 89.6%; and 92.6%, respectively. Mean intergrader agreements for assessment of overall Unusable, Usable and Excellent 6x6-mm OCT-A images (21 images) were 73.3 %, 57.1% and 83.8%, agreements for Unusable VD and FAZ area were 60.0 % and 84.8 %, respectively. Three analytic scenarios were developed to account for common types of bias related to suboptimal OCT-A image quality and ocular comorbidities. Conclusion These recommendations provide a framework for working with OCT-A imaging in population studies. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement Funding Sources The Maastricht Study This study was supported by the European Regional Development Fund via OP-Zuid, the Province of Limburg, the Dutch Ministry of Economic Affairs (grant 31O.041), Stichting De Weijerhorst (Maastricht, the Netherlands), the Pearl String Initiative Diabetes (Amsterdam, the Netherlands), the Cardiovascular Center (CVC, Maastricht, the Netherlands), CARIM School for Cardiovascular Diseases (Maastricht, the Netherlands), CAPHRI School for Public Health and Primary Care (Maastricht, the Netherlands), NUTRIM School for Nutrition and Translational Research in Metabolism (Maastricht, the Netherlands), Stichting Annadal (Maastricht, the Netherlands), Health Foundation Limburg (Maastricht, the Netherlands), Perimed (Jarfalla, Sweden), and by unrestricted grants from Janssen-Cilag B.V. (Tilburg, the Netherlands), Novo Nordisk Farma B.V. (Alphen aan den Rijn, the Netherlands), and Sanofi-Aventis Netherlands B.V. (Gouda, the Netherlands). Rotterdam Study The Rotterdam Study is supported by the Algemene Nederlandse Vereniging ter Voorkoming van Blindheid, Oogfonds, Stichting voor Ooglijders, Stichting voor Blindenhulp, Henkes stichting, Rotterrdams Stichting voor Blindenbelangen, and Landelijke Stichting voor Blinden en Slechtzienden. Additional support was given by the Erasmus Medical Center, Erasmus University, Netherlands Organization for the Health Research and Development (ZonMw), the Research Institute for Diseases in the Elderly, the Ministry of Education, Culture and Science, the Ministry for Health, Welfare and Sports, the European Commission (DG XII), and the Municipality of Rotterdam. Rhineland Study The Rhineland Study (P.I. Breteler) is primarily supported by DZNE core funding. The DZNE is funded by the Federal Ministry of Education and Research (BMBF) and the Ministry of Culture and Science of the German State of North Rhine-Westphalia. Framingham Heart Study National Institutes of Health R01AG066524 (AHK, SS), FHS contract 75N92019D00031, and P30AG066546. SIGNATR This work was supported by: National Eye Institute under R01 EY027134 and Government of India Department of Biotechnology under Grant BT/PR22701/MED/15/166/2016. ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: Ethics committee/IRB of all institutions (Maastricht University, Rotterdam University, Framingham Heart Study, Rhineland Study, and Rothschild Hospital) gave ethical approval for this work I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes All data produced in the present study are available upon reasonable request to the authors

Diabetic retinopathy (DR), a prevalent microvascular complication of diabetes, is the fifth leading cause of blindness worldwide. Given the critical nature of the disease, it is paramount that individuals with diabetes undergo annual screening for early and timely detection of DR, facilitating prompt ophthalmic assessment and intervention. However, screening for DR, which involves assessing visual acuity and retinal examination through ophthalmoscopy or retinal photography, presents a significant global challenge due to the massive volume of individuals requiring annual reviews. To counter this challenge, there has been an increasing interest in the potential of artificial intelligence (AI) tools for automated diagnosis of DR. The AI tools primarily utilize deep learning (DL) techniques and are tailored to analyse extensive medical image data and provide diagnostic outputs, essentially streamline the DR screening process. However, the development of such AI tools requires access to a comprehensive retinal image database with a plethora of high-resolution fundus images from various cameras, covering all DR lesions. Additionally, the accurate training of these AI algorithms necessitates skilled professionals, such as optometrists or ophthalmologists, to provide reliable ground truths that ensure the precision of the diagnostic outputs. To address these prerequisites, we have initiated a study involving multiple institutions to establish a large-scale online 'Retinal Image Database’ in India, aiming to contribute significantly to DR research. This paper delineates the methodology employed for this significant undertaking, detailing the steps taken to create the large retinal image database, as well as the framework for developing a cost-effective, robust AI-based DR diagnostic tool. Our work is expected to mark a significant stride in DR detection and management, promising a more efficient and scalable solution for tackling this global health challenge.

Meta-analysis is a popular technique for aggregating evidence from a variety of similar studies for collective comparison. This paper presents data on genes and genetic variants (SNPs) associated with Type 2 Diabetes Mellitus, Diabetic Retinopathy, and Diabetic Nephropathy. Retinopathy and nephropathy are microvascular diseases that occur as a consequence of long-term diabetes. An overlap of the genes and SNPs revealed only one gene and one SNP to be common among the three diseases. The pathways associated with these diseases showed overlap to a certain extent. In this study, we have pooled all the genes and genetic variants associated with these three diseases and analyzed the overlaps/interactions. Such analyses lead to a better understanding of the disease mechanisms. We have analyzed the data using various tools such as KEGG, GO, and Network Analyst. Several genes were identified that have a significant role in the pathways of all three diseases. EPO is the only gene which was identified to be associated commonly among the three diseases. Among the genetic variants, rs1617640 of the gene EPO is the only common variant among the three diseases. The data cataloged in this paper serves as a genomic resource for diabetes and its associated microvascular diseases and it will also benefit as a resource for scientists working in this area of research.

OBJECTIVE:Vascular endothelial growth factor (VEGF) plays a key role in diabetic retinopathy (DR). Previously, we have reported an association between mutations in a gene coding for the L-type calcium channel subunit, VEGF and DR. L-type calcium channel blockers (LTCCBs) have been widely used as antihypertensive medication (AHM), but their association with VEGF and DR is still unclear. Therefore, we explored the effect of LTCCBs compared to other AHMs on VEGF concentrations in retinal cells and human serum. Furthermore, we evaluated the association between the use of LTCCBs and the risk of severe diabetic eye disease (SDED). RESEARCH DESIGN AND METHODS:Müller cells (MIO-M1) were cultured as per recommended protocol and treated with LTCCBs and other AHMs. VEGF secreted from cells were collected at 24 hours intervals. In an interventional study, 39 individuals received LTCCBs or other AHM for four weeks with a four-week wash-out placebo period between treatments. VEGF was measured during the medication and placebo periods. Finally, we evaluated the risk of SDED associated with LTCCB usage in 192 individuals from the FinnDiane Study in an observational setting. RESULTS:In the cell cultures, the medium VEGF concentration increased time-dependently after amlodipine (P<0.01) treatment, but not after losartan (P>0.01), or lisinopril (P>0.01). Amlodipine, but no other AHM, increased the serum VEGF concentration (P<0.05) during the interventional clinical study. The usage of LTCCB was not associated with the risk of SDED in the observational study. CONCLUSIONS:LTCCB increases VEGF concentrations in retinal cells and human serum. However, the usage of LTCCBs does not appear to be associated with SDED in adults with type 1 diabetes.

Purpose: Inherited retinal dystrophies (IRD) are a heterogeneous group of retinal diseases leading to progressive loss of photoreceptors through apoptosis. Retinitis pigmentosa (RP) is considered the most common form of IRD. Panel-based testing in RP has proven effective in identifying the causative genetic mutations in 70% and 80% of the patients. This is a retrospective, observational, single-center study of 107 RP patients who had undergone next-generation sequencing-based targeted gene panel testing for IRD genes. These patients were inspected for common phenotypic features to arrive at meaningful genotype–phenotype correlation. Methods: Patients underwent complete ophthalmic examination, and blood was collected from the proband for DNA extraction after documenting the pedigree. Targeted Next Generation Sequencing (NGS) was done by panel-based testing for IRD genes followed by co-segregation analysis wherever applicable. Results: Of the 107 patients, 72 patients had pathogenic mutations. The mean age of onset of symptoms was 14 ± 12 years (range: 5–55). Mean (Best Corrected Visual Acuity) BCVA was 6/48 (0.9 logMAR) (range 0.0–3.0). At presentation, over one-third of eyes had BCVA worse than 6/60 (<1 logMAR). Phenotype analysis with the gene defects showed overlapping features, such as peripheral well-defined chorioretinal atrophic patches in patients with CERKL , PROM1 , and RPE65 gene mutations and large macular lesions in patients with RDH12 and CRX gene mutations, respectively. Nummular or clump-like pigmentation was noted in CRB1 , TTC8 , PDE6A , and PDE6B . Conclusion: NGS-based genetic testing can help clinicians to diagnose RP more accurately, and phenotypic correlations can also help in better patient counselling with respect to prognosis and guidance regarding ongoing newer gene-based therapies.

Purpose: The purpose of the South Indian GeNetics of DiAbeTic Retinopathy (SIGNATR) Study is to identify non-genetic and genetic risk factors associated with diabetic retinopathy (DR). This report examines the non-genetic risk factors for DR in South Indian patients. Methods: Participants with South Indian ancestry and type 2 diabetes (T2D) were included from two sources: the Sankara Nethralaya Diabetic Retinopathy and Molecular Genetics Study (SN-DREAMS) and prospective recruitment at Sankara Nethralaya affiliates. Fundus photography and optical coherence tomography (OCT) were obtained on participants. Fundus images were graded for DR severity and OCTs were graded for center-involved diabetic macular edema (ciDME). Multivariate analyses were performed using stepwise logistic regression to assess effects of the demographic and clinical factors on proliferative DR (PDR) and DME. Results: Among the 2941 participants with DR grading, participants with PDR were more likely to be younger [odds ratio (OR)=0.95], men (OR = 1.83), have a longer duration of diabetes (OR = 1.10), have a higher hemoglobin A1c (OR = 1.12), have albuminuria (OR = 5.83), have hypertension (OR = 1.69), have a higher HDL (OR = 1.02) and a lower total cholesterol (OR = 0.99) (all p < 0.05). Among the 483 participants with gradable OCT scans, participants who had ciDME were more likely to be younger (OR = 0.97), men (OR = 2.80), have a longer duration of diabetes (OR = 1.06), have lower triglycerides (OR = 0.99), and have albuminuria (OR = 3.12) (all p < 0.05). Conclusions: Younger age, male sex, longer duration of diabetes, higher HbA1c, and presence of albuminuria were identified as risk factors for PDR and DME in a South Indian population with T2D.

Age-related macular degeneration (AMD) and polypoidal choroidal vasculopathy (PCV) are sister diseases and have several similar clinical features and still have few genetic differences. The association of HERPUD1 (homocysteine inducible ER protein with ubiquitin like domain 1) gene variant rs2217332 with PCV is known; however, such association with AMD has not been reported in the Indian population. We analyzed the association of rs2217332 with PCV and AMD to identify the preferential association of this variant with these diseases. This is a population-based case–control study consisting of 422 patients (129 AMD cases; 101 PCV cases, 192 healthy controls) recruited from the vitreoretinal clinic Sankara Nethralaya. The sample size for the study was calculated using appropriate power calculation methods. Genotype was determined using PCR-based Sanger sequencing. The SPSS V23.0 statistical package tool was used to calculate chi-square and ROC to determine the association of rs2217332 with control, AMD, and PCV. Here, we report for the first time the association of this genetic variant (rs2217332) with AMD and PCV in the Indian population. The case–control study shows a significant association of this SNP with PCV (P value = 0.002); however, this variant is not significantly associated with AMD (P value = 0.602). Comparison between AMD (as control) and PCV (as case) also showed significant association of the SNP with PCV (P value = 0.02). Minor allele A conferred to increase the risk of PCV. The study concludes that the genetic variant rs2217332 in HERPUD1 gene is highly significantly associated with PCV and not with AMD in Indian populations.

Zebrafish, a popular organism for studying embryonic development and for modeling human diseases, has so far lacked a systematic functional annotation program akin to those in other animal models. To address this, we formed the international DANIO-CODE consortium and created a central repository to store and process zebrafish developmental functional genomic data. Our data coordination center ( https://danio-code.zfin.org ) combines a total of 1,802 sets of unpublished and re-analyzed published genomic data, which we used to improve existing annotations and show its utility in experimental design. We identified over 140,000 cis-regulatory elements throughout development, including classes with distinct features dependent on their activity in time and space. We delineated the distinct distance topology and chromatin features between regulatory elements active during zygotic genome activation and those active during organogenesis. Finally, we matched regulatory elements and epigenomic landscapes between zebrafish and mouse and predicted functional relationships between them beyond sequence similarity, thus extending the utility of zebrafish developmental genomics to mammals.

Purpose: Stickler syndrome is associated with the development of rhegmatogenous retinal detachment (RRD), and often presents with ocular, auditory, skeletal, and orofacial abnormalities. Molecular analysis has proven effective in diagnosis, confirmation and classification of the disease. We aimed to describe the utility of next-generation sequencing (NGS) in genetic analysis of four Indian families with suspected Stickler syndrome. Methods: The index cases presented with retinal detachment with family history. Genetic analysis in the index case was performed by next-generation sequencing of inherited retinal degeneration genes, and validated by Sanger sequencing followed by co-segregation analysis in the other family members. Results: Twenty patients were included for the genetic analysis (15 males and 5 females from four families). Clinical details were available for 15 patients (30 eyes). Fourteen eyes (11 patients) developed RRD. In the 16 eyes without RRD, 8 underwent barrage laser to lattice degeneration and 8 were under observation. Disease segregating heterozygous mutations with pathogenic/likely pathogenic effect was identified in COL2A1 (c.4318-1G>A, c.141G>A, c.1221+1G>A for 3 families) and COL11A1 (c.1737+1 G>A for 1 family) gene. In addition to the mutation in the COL2A1 gene, a pathogenic heterozygous variant associated with risk for arrhythmogenic right ventricular cardiomyopathy (ARVC) was identified in one member. Conclusion: NGS testing confirmed the presence of the causative gene for Stickler syndrome in the index case followed by evaluation of family members and confirmation of genetic and ocular findings. We believe that this may be the first such report of families with RRD from India.

Dear Editior, Diabetic retinopathy (DR) is the most common microvascular ocular complication of diabetes due to prolonged uncontrolled hyperglycemia. The population-based studies in India reported a prevalence of DR were 18% and 10% in urban and rural areas respectively.[12] Nearly 50-70% of DR related visual complications can be prevented by timely screening and intervention.[3] To improve the yield of DR screening, we did a targeted DR screening camp for people diagnosed as diabetes for at least more than 10 years. We planned to reward people who despite a longer duration of diabetes didn‘t have any DR by a certificate and medal, few were asked to share their views on practices they followed to control diabetes [Fig. 1]. The announcement for screening was made to public with the help of social media and by other modes [Fig. 2]. Nearly 84 people participated in the screening. The average age was 66.47 ± 10.77 years of which 66 (68.57%) were males. The mean duration of diabetes was 16.96 ± 9.12 years. Of the people who came for screening, 70 (85.36%) people were identified with normal fundus.Figure 1: Camp site for diabetic retinopathy screeningFigure 2: Source of information for diabetic retinopathy screeningPeople with no DR were asked to share about the measures they take in their life. Majority reported following a diabetes diet, medicines on time, followed by regular exercise like walking and yoga as a reason for good glycemic control. Few selective quotes are mentioned below. “Doing exercise will prevent the early onset of diabetes for the younger age people”. “I feel the reason for onset of diabetes is life style changes and food habits apart from genetic factors. I maintain my diabetes under control by regular walking and following diet”. Best means of communication to reach target population seems to be SMS messaging and word of mouth by a known person. A component of reward motivates them to continue their good glycemic control. Financial support and sponsorship This study was supported by the Lions Club International Foundation (SFP2050/UND). Conflicts of interest There are no conflicts of interest.

Zebrafish, a popular model for embryonic development and for modelling human diseases, has so far lacked a systematic functional annotation programme akin to those in other animal models. To address this, we formed the international DANIO-CODE consortium and created the first central repository to store and process zebrafish developmental functional genomic data. Our Data Coordination Center ( https://danio-code.zfin.org ) combines a total of 1,802 sets of unpublished and reanalysed published genomics data, which we used to improve existing annotations and show its utility in experimental design. We identified over 140,000 cis-regulatory elements in development, including novel classes with distinct features dependent on their activity in time and space. We delineated the distinction between regulatory elements active during zygotic genome activation and those active during organogenesis, identifying new aspects of how they relate to each other. Finally, we matched regulatory elements and epigenomic landscapes between zebrafish and mouse and predict functional relationships between them beyond sequence similarity, extending the utility of zebrafish developmental genomics to mammals.

Purpose: Genetic testing for primary mutations m.3460G>A, m.11778G>A, and m.14484T>C in ND1, ND4, and ND6 genes of mitochondrial DNA is the recommended assay for Leber hereditary optic neuropathy (LHON; OMIM 535000). This report discusses the outcome of molecular genetic screening for these three primary mutations in suspected LHON cases in India. Methods: Two hundred and seventy-eight unrelated presumed LHON patients who were seen at the neuro-ophthalmology clinic of a tertiary eye care center from 2014-2018 were analyzed. They were genotyped for the three common variants by polymerase chain reaction-based direct sequencing, and their plasmy status was also determined by restriction enzyme digestion. Results: Eighty two of 278 patients were positive for one of the 3 common mutations with m.11778G>A in ND4 gene more frequently distributed (N=72) in homoplasmic state (N=59/82). The mean onset age of visual loss was 21.1years (SD, 9.8 years; range, 5-58 years) in patients harboring the primary mutation. The most common clinical presentation was bilateral sequential painless vision loss with central and cecocentral scotomas in the visual field due to optic disc atrophy. Conclusions: The study subjects are a sample of a much larger number of suspected LHON cases tested for primary mutations in India. (N= 278) and 29.4% (82/278) of patients harbour one of the 3 common mutations. Screening the entire mitochondrial genome and the other nuclear genes encoding mitochondrial protein, would probably aid in identifying the other less common mtDNA mutations causing LHON in Indian population.

Leber congenital amaurosis (LCA) is a severe autosomal recessive retinal degenerative disease. The current study describes exome sequencing results for two unrelated Indian LCA patients carrying novel nonsense p.(Glu636*) and frameshift p.(Pro2281Leufs*63) mutations in the ALMS1 gene. Although ALMS1 gene mutations are associated with Alstrom syndrome (AS), the current patients did not exhibit typical syndromic features of AS. These data suggest that ALMS1 should be included in the candidate gene panel for LCA to improve diagnostic efficiency.

The study of translational regulation requires reliable measurement of both mRNA levels and protein synthesis. Cytoplasmic polyadenylation is a prevalent mode of translational regulation during oogenesis and early embryogenesis. Here the length of the poly(A) tail of an mRNA is coupled to its translatability. We describe a protocol to identify translationally regulated genes and measure their translation rate in the early zebrafish embryo using genome-wide polysome profiling. This protocol relies on the isolation of mRNA by means of an rRNA depletion strategy, which avoids capture bias due to short poly(A) tail that can occur when using conventional oligo(dT)-based methods. We also present a simple PCR-based method to measure the poly(A) tail length of selected mRNAs.

Retinal detachment (RD) is an ocular emergency, which needs quick intervention to preclude permanent vision loss. In general, ocular ultrasound is used by ophthalmologists to enhance their judgment in detecting RD in eyes with media opacities which precludes the retinal evaluation. However, the quality of ultrasound (US) images may be degraded due to the presence of noise, and other retinal conditions may cause membranous echoes. All these can influence the accuracy of diagnosis. Hence, to overcome the above, we are proposing an automated system to detect RD using texton, higher order spectral (HOS) cumulants and locality sensitive discriminant analysis (LSDA) techniques. Our developed method is able to classify the posterior vitreous detachment and RD using support vector machine classifier with highest accuracy of 99.13%. Our system is ready to be tested with more diverse ultrasound images and aid ophthalmologists to arrive at a more accurate diagnosis.

Significance Bone remodeling requires a balanced interplay of osteoblasts and osteoclasts. While the intercellular signaling that triggers bone cell differentiation is well understood, it remains unclear how bone progenitor cells are recruited to remodeling sites. Various chemokines are upregulated under osteoporotic conditions. However, whether they are involved in progenitor recruitment or instead have inflammatory roles is unknown. Here we used a medaka fish osteoporosis model to identify the chemokine ligand Cxcl9l and receptor Cxcr3.2 as essential to control osteoclast progenitor recruitment and differentiation at bone resorption sites. Cxcr3.2 activity can be blocked by small-molecule inhibitors that protect bone from osteoporotic insult. Our study demonstrates the potential of fish for osteoporosis drug discovery and opens avenues for future osteoporosis therapy.

Background and objectivesPolypoidal choroidal vasculopathy (PCV) is a retinal disorder characterized by the presence of aneurismal polypoidal lesions in the choroidal vasculature. A single nucleotide polymorphism (SNP) is a common genetic variant which may be associated with the disease. This study is to investigate the association of HERPUD1 (rs2217332) gene with PCV in the Indian population and develop an automated system for genotype and phenotype correlation using fundus images and machine learning methods.MethodsA cohort of 54 PCV patients and 120 control subjects were recruited for the study. Genotyping of SNP (HERPUD1, rs2217332) was performed by following polymerase chain reaction and direct sequencing method. Statistical association of SNP to PCV was determined using chi-square analysis. The acquired GG and AG images were preprocessed using an adaptive histogram. 19 and 18 texture features were extracted from the images in the PCV naïve cases and PCV patients on treatment, respectively. Student's independent t-test was then employed for the selection of significant features, which were input to the ensemble tree for automated classification. Leave-one-out validation was used to evaluate the system.ResultsHERPUD1 rs2217332 SNP is significantly associated in PCV patients compared to control (P = 0.0296, odds ratio [OD] = 2.297, 95% confidence interval [CI] = 1.087–4.856) in the Indian population. High F1 and precision values of 85.71%, 86.84% and 85.71%, 93.75% were achieved in the pre and post- treatment phases, respectively.ConclusionOur results suggest that the HERPUD1 polymorphism is associated in PCV patients. Based on our analysis, it may be possible to predict the genotype and disease status of PCV patients using fundus images in assistance with a machine learning algorithm.