TMEM16A partners with mTOR to influence pathways of cell survival, proliferation, and migration in cholangiocarcinoma.

Expression of transmembrane protein 16 A (TMEM16A), a calcium activated chloride channel, is elevated in some human cancers and impacts tumor cell proliferation, metastasis, and patient outcome. Evidence presented here uncovers a molecular synergy between TMEM16A and mechanistic/mammalian target of rapamycin (mTOR), a serine-threonine kinase that is known to promote cell survival and proliferation in cholangiocarcinoma (CCA), a lethal cancer of the secretory cells of bile ducts. Analysis of gene and protein expression in human CCA tissue and CCA cell line detected elevated TMEM16A expression and Cl- channel activity. The Cl- channel activity of TMEM16A impacted the actin cytoskeleton and the ability of cells to survive, proliferate, and migrate as revealed by pharmacological inhibition studies. The basal activity of mTOR, too, was elevated in the CCA cell line compared with the normal cholangiocytes. Molecular inhibition studies provided further evidence that TMEM16A and mTOR were each able to influence the regulation of the other's activity or expression respectively. Consistent with this reciprocal regulation, combined TMEM16A and mTOR inhibition produced a greater loss of CCA cell survival and migration than their individual inhibition alone. Together these data reveal that the aberrant TMEM16A expression and cooperation with mTOR contribute to a certain advantage in CCA.NEW & NOTEWORTHY This study points to the dysregulation of transmembrane protein 16 A (TMEM16A) expression and activity in cholangiocarcinoma (CCA), the inhibition of which has functional consequences. Dysregulated TMEM16A exerts an influence on the regulation of mechanistic/mammalian target of rapamycin (mTOR) activity. Moreover, the reciprocal regulation of TMEM16A by mTOR demonstrates a novel connection between these two protein families. These findings support a model in which TMEM16A intersects the mTOR pathway to regulate cell cytoskeleton, survival, proliferation, and migration in CCA.

Assistive Framework for Automatic Detection of All the Zones in Retinopathy of Prematurity Using Deep Learning.

Retinopathy of prematurity (ROP) is a potentially blinding disorder seen in low birth weight preterm infants. In India, the burden of ROP is high, with nearly 200,000 premature infants at risk. Early detection through screening and treatment can prevent this blindness. The automatic screening systems developed so far can detect "severe ROP" or "plus disease," but this information does not help schedule follow-up. Identifying vascularized retinal zones and detecting the ROP stage is essential for follow-up or discharge from screening. There is no automatic system to assist these crucial decisions to the best of the authors' knowledge. The low contrast of images, incompletely developed vessels, macular structure, and lack of public data sets are a few challenges in creating such a system. In this paper, a novel method using an ensemble of "U-Network" and "Circle Hough Transform" is developed to detect zones I, II, and III from retinal images in which macula is not developed. The model developed is generic and trained on mixed images of different sizes. It detects zones in images of variable sizes captured by two different imaging systems with an accuracy of 98%. All images of the test set (including the low-quality images) are considered. The time taken for training was only 14 min, and a single image was tested in 30 ms. The present study can help medical experts interpret retinal vascular status correctly and reduce subjective variation in diagnosis.

Metabolic and oncogenic adaptations to pyruvate dehydrogenase inactivation in fibroblasts.

Eukaryotic cell metabolism consists of processes that generate available energy, such as glycolysis, the tricarboxylic acid (TCA) cycle, and oxidative phosphorylation (Oxphos), and those that consume it, including macromolecular synthesis, the maintenance of ionic gradients, and cellular detoxification. By converting pyruvate to acetyl-CoA (AcCoA), the pyruvate dehydrogenase (PDH) complex (PDC) links glycolysis and the TCA cycle. Surprisingly, disrupting the connection between glycolysis and the TCA cycle by inactivation of PDC has only minor effects on cell replication. However, the molecular basis for this metabolic re-equilibration is unclear. We report here that CRISPR/Cas9-generated PDH-knockout (PDH-KO) rat fibroblasts reprogrammed their metabolism and their response to short-term c-Myc (Myc) oncoprotein overexpression. PDH-KO cells replicated normally but produced surprisingly little lactate. They also exhibited higher rates of glycolysis and Oxphos. In addition, PDH-KO cells showed altered cytoplasmic and mitochondrial pH, redox states, and mitochondrial membrane potential (ΔΨM). Conditionally activated Myc expression affected some of these parameters in a PDH-dependent manner. PDH-KO cells had increased oxygen consumption rates in response to glutamate, but not to malate, and were depleted in all TCA cycle substrates between α-ketoglutarate and malate despite high rates of glutaminolysis, as determined by flux studies with isotopically labeled glutamine. Malate and pyruvate were diverted to produce aspartate, thereby potentially explaining the failure to accumulate lactate. We conclude that PDH-KO cells maintain proliferative capacity by utilizing glutamine to supply high rates of AcCoA-independent flux through the bottom portion of the TCA cycle while accumulating pyruvate and aspartate that rescue their redox defects.

ß-Catenin mutations as determinants of hepatoblastoma phenotypes in mice.

Hepatoblastoma (HB) is the most common pediatric liver cancer. Although long-term survival of HB is generally favorable, it depends on clinical stage, tumor histology, and a variety of biochemical and molecular features. HB appears almost exclusively before the age of 3 years, is represented by seven histological subtypes, and is usually associated with highly heterogeneous somatic mutations in the catenin ß1 (CTNNB1) gene, which encodes ß-catenin, a Wnt ligand-responsive transcriptional co-factor. Numerous recurring ß-catenin mutations, not previously documented in HB, have also been identified in various other pediatric and adult cancer types. Little is known about the underlying factors that determine the above HB features and behaviors or whether non-HB-associated ß-catenin mutations are tumorigenic when expressed in hepatocytes. Here, we investigated the oncogenic properties of 14 different HB- and non-HB-associated ß-catenin mutants encoded by Sleeping Beauty vectors following their delivery into the mouse liver by hydrodynamic tail-vein injection. We show that all ß-catenin mutations, as well as WT ß-catenin, are tumorigenic when co-expressed with a mutant form of yes-associated protein (YAP). However, tumor growth rates, histologies, nuclear-to-cytoplasmic partitioning, and metabolic and transcriptional landscapes were strongly influenced by the identities of the ß-catenin mutations. These findings provide a context for understanding at the molecular level the notable biological diversity of HB.

Myc and ChREBP transcription factors cooperatively regulate normal and neoplastic hepatocyte proliferation in mice.

Analogous to the c-Myc (Myc)/Max family of bHLH-ZIP transcription factors, there exists a parallel regulatory network of structurally and functionally related proteins with Myc-like functions. Two related Myc-like paralogs, termed MondoA and MondoB/carbohydrate response element-binding protein (ChREBP), up-regulate gene expression in heterodimeric association with the bHLH-ZIP Max-like factor Mlx. Myc is necessary to support liver cancer growth, but not for normal hepatocyte proliferation. Here, we investigated ChREBP's role in these processes and its relationship to Myc. Unlike Myc loss, ChREBP loss conferred a proliferative disadvantage to normal murine hepatocytes, as did the combined loss of ChREBP and Myc. Moreover, hepatoblastomas (HBs) originating in myc-/-, chrebp-/-, or myc-/-/chrebp-/- backgrounds grew significantly more slowly. Metabolic studies on livers and HBs in all three genetic backgrounds revealed marked differences in oxidative phosphorylation, fatty acid ß-oxidation (FAO), and pyruvate dehydrogenase activity. RNA-Seq of livers and HBs suggested seven distinct mechanisms of Myc-ChREBP target gene regulation. Gene ontology analysis indicated that many transcripts deregulated in the chrebp-/- background encode enzymes functioning in glycolysis, the TCA cycle, and ß- and ω-FAO, whereas those dysregulated in the myc-/- background encode enzymes functioning in glycolysis, glutaminolysis, and sterol biosynthesis. In the myc-/-/chrebp-/- background, additional deregulated transcripts included those involved in peroxisomal ß- and α-FAO. Finally, we observed that Myc and ChREBP cooperatively up-regulated virtually all ribosomal protein genes. Our findings define the individual and cooperative proliferative, metabolic, and transcriptional roles for the "Extended Myc Network" under both normal and neoplastic conditions.

Sequential adaptive changes in a c-Myc-driven model of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a common cancer that frequently overexpresses the c-Myc (Myc) oncoprotein. Using a mouse model of Myc-induced HCC, we studied the metabolic, biochemical, and molecular changes accompanying HCC progression, regression, and recurrence. These involved altered rates of pyruvate and fatty acid ß-oxidation and the likely re-directing of glutamine into biosynthetic rather than energy-generating pathways. Initial tumors also showed reduced mitochondrial mass and differential contributions of electron transport chain complexes I and II to respiration. The uncoupling of complex II's electron transport function from its succinate dehydrogenase activity also suggested a mechanism by which Myc generates reactive oxygen species. RNA sequence studies revealed an orderly progression of transcriptional changes involving pathways pertinent to DNA damage repair, cell cycle progression, insulin-like growth factor signaling, innate immunity, and further metabolic re-programming. Only a subset of functions deregulated in initial tumors was similarly deregulated in recurrent tumors thereby indicating that the latter can "normalize" some behaviors to suit their needs. An interactive and freely available software tool was developed to allow continued analyses of these and other transcriptional profiles. Collectively, these studies define the metabolic, biochemical, and molecular events accompanyingHCCevolution, regression, and recurrence in the absence of any potentially confounding therapies.

TMEM16A/ANO1 suppression improves response to antibody-mediated targeted therapy of EGFR and HER2/ERBB2.

TMEM16A, a Ca2+ -activated Cl- channel, contributes to tumor growth in breast cancer and head and neck squamous cell carcinoma (HNSCC). Here, we investigated whether TMEM16A influences the response to EGFR/HER family-targeting biological therapies. Inhibition of TMEM16A Cl- channel activity in breast cancer cells with HER2 amplification induced a loss of viability. Cells resistant to trastuzumab, a monoclonal antibody targeting HER2, showed an increase in TMEM16A expression and heightened sensitivity to Cl- channel inhibition. Treatment of HNSCC cells with cetuximab, a monoclonal antibody targeting EGFR, and simultaneous TMEM16A suppression led to a pronounced loss of viability. Biochemical analyses of cells subjected to TMEM16A inhibitors or expressing chloride-deficient forms of TMEM16A provide further evidence that TMEM16A channel function may play a role in regulating EGFR/HER2 signaling. These data demonstrate that TMEM16A regulates EGFR and HER2 in growth and survival pathways. Furthermore, in the absence of TMEM16A cotargeting, tumor cells may acquire resistance to EGFR/HER inhibitors. Finally, targeting TMEM16A improves response to biological therapies targeting EGFR/HER family members.

Coordinated Activities of Multiple Myc-dependent and Myc-independent Biosynthetic Pathways in Hepatoblastoma.

Hepatoblastoma (HB) is associated with aberrant activation of the ß-catenin and Hippo/YAP signaling pathways. Overexpression of mutant ß-catenin and YAP in mice induces HBs that express high levels of c-Myc (Myc). In light of recent observations that Myc is unnecessary for long-term hepatocyte proliferation, we have now examined its role in HB pathogenesis using the above model. Although Myc was found to be dispensable for in vivo HB initiation, it was necessary to sustain rapid tumor growth. Gene expression profiling identified key molecular differences between myc+/+ (WT) and myc-/- (KO) hepatocytes and HBs that explain these behaviors. In HBs, these included both Myc-dependent and Myc-independent increases in families of transcripts encoding ribosomal proteins, non-structural factors affecting ribosome assembly and function, and enzymes catalyzing glycolysis and lipid bio-synthesis. In contrast, transcripts encoding enzymes involved in fatty acid ß-oxidation were mostly down-regulated. Myc-independent metabolic changes associated with HBs included dramatic reductions in mitochondrial mass and oxidative function, increases in ATP content and pyruvate dehydrogenase activity, and marked inhibition of fatty acid ß-oxidation (FAO). Myc-dependent metabolic changes included higher levels of neutral lipid and acetyl-CoA in WT tumors. The latter correlated with higher histone H3 acetylation. Collectively, our results indicate that the role of Myc in HB pathogenesis is to impose mutually dependent changes in gene expression and metabolic reprogramming that are unattainable in non-transformed cells and that cooperate to maximize tumor growth.

Minimally fibrotic stage 5 ROP: a clinical prognostic factor in eyes undergoing vitrectomy for stage 5 retinopathy of prematurity.

TITLE: Minimally fibrotic Stage 5: A clinical prognostic factor in eyes undergoing vitrectomy for stage 5 retinopathy of prematurity (ROP). PURPOSE: To define minimally fibrotic stage 5 ROP and to demonstrate better surgical outcomes in this subgroup. METHODS: A cohort of eligible eyes with stage 5 ROP undergoing vitrectomy were further divided into those with minimally fibrotic stage 5 and others. Minimally fibrotic stage 5 ROP was defined as those stage 5 eyes having a translucent retrolental membrane with visibility of the underlying retinal vessels and absence of anteriorly rotated ciliary processes. We reported on anatomic and visual outcomes in eyes with defined characteristics of this subgroup identified before surgery and then observing them post vitrectomy over a period of time as compared to others. RESULTS: Minimally fibrotic stage 5 eyes had better visual (p = .0068) and anatomical outcomes (p = .0001). CONCLUSION: Minimally fibrotic stage 5 ROP shows ridge to ridge traction that has resulted in a traction retinal detachment with a retrolental membrane, but fibrotic change has just begun. This stage 5 ROP with limited fibrosis represents a subset of this stage 5 ROP with a positive anatomic and visual prognosis.

Vision development over an extended follow-up period in babies after successful vitrectomy for stage 4b retinopathy of prematurity.

PURPOSE/BACKGROUND: To demonstrate improvement in the vision of babies after successful vitrectomy for stage 4b retinopathy of prematurity (ROP) over an extended period of time. METHODS: This was an observational prospective case series. Eight babies who had undergone successful vitrectomy in either their only seeing eye (or both eyes) with stage 4b ROP were followed up post-operatively for a period of 80 weeks or more. Vision with Teller acuity chart, refraction, binocular indirect ophthalmoscopy, and documentation with RetCam was done at each visit. Vision of the (only/better) seeing operated eye with corrective glasses was graded for the purpose of statistical evaluation. Paired t test was performed to compare the vision prior to 30 weeks and at or after 80 weeks. RESULTS: Statistically significant improvement in vision was noted at or after 80 weeks as compared to the vision recorded before 30 weeks (p = 0.0062). CONCLUSIONS: Unlike in adult intraocular surgeries where stable visual acuity is reached well before 30 weeks, continuing improvement at 80 weeks and beyond is noted. Gradual restoration of the retinal architecture and plasticity of the infant's developing brain are thought to be responsible.

HVDROPDB datasets for research in retinopathy of prematurity.

Retinopathy of prematurity (ROP) is a retinal disorder that may bring about blindness in preterm infants. Early detection and treatment of ROP can prevent this blindness. The gold standard technique for ROP screening is indirect ophthalmoscopy performed by ophthalmologists. The scarcity of medical professionals and inter-observer heterogeneity in ROP grading are two of the screening concerns. Researchers employ artificial intelligence (AI) driven ROP screening systems to assist medical experts. A major hurdle in developing these systems is the unavailability of annotated data sets of fundus images. Anatomical landmarks in the retina, such as the optic disc, macula, blood vessels, and ridge, are used to identify ROP characteristics. HVDROPDB is the first dataset to be published for the retinal structure segmentation of fundus images of preterm infants. It is prepared from two diverse imaging systems on the Indian population for segmenting the lesions mentioned above and annotated by a group of ROP experts. Each dataset contains retinal fundus images of premature infants with the ground truths prepared manually to assist researchers in developing explainable automated screening systems.

Natural course of non-center-involving diabetic macular edema progression in patients under initial observation.

PURPOSE: We aim to report the natural course of non-center involving diabetic macular edema (NCIDME) progression to center involving diabetic macular edema (CIDME) and associated risk factors. METHODS: This is a multicenter retrospective comparative study. Data was collected from electronic medical records from 8 centers in India covering. We included patients with type 2 diabetes above 18 years of age with treatment-naïve NCIDME on OCT and best-corrected visual acuity at baseline of 6/12 or better who were under observation for NCIDME and had 2 years follow-up data. RESULTS: Out of 72 patients with NCIDME, 26.38% patients progressed to CI DME by 2 years, and the visit wise proportion was 11.11% at 6 months, 7% at 1st year and 8.3% at 2 years. The change in CST was statistically significant at 2 years in patients who developed CIDME, the mean difference was 137.73 ± 48.56 microns p = 0.045. Duration of diabetes mellitus > 10 years was the only risk factor for conversion to CIDME. CONCLUSION: A quarter of eyes with NCIDME developed CIDME and 15% progressed from NPDR to PDR by 2 years, highlighting the disease burden in these patients with NCIDME.

Hydrolysis of aromatic ß-glucosides by non-pathogenic bacteria confers a chemical weapon against predators.

Bacteria present in natural environments such as soil have evolved multiple strategies to escape predation. We report that natural isolates of Enterobacteriaceae that actively hydrolyze plant-derived aromatic ß-glucosides such as salicin, arbutin and esculin, are able to avoid predation by the bacteriovorous amoeba Dictyostelium discoideum and nematodes of multiple genera belonging to the family Rhabditidae. This advantage can be observed under laboratory culture conditions as well as in the soil environment. The aglycone moiety released by the hydrolysis of ß-glucosides is toxic to predators and acts via the dopaminergic receptor Dop-1 in the case of Caenorhabditis elegans. While soil isolates of nematodes belonging to the family Rhabditidae are repelled by the aglycone, laboratory strains and natural isolates of Caenorhabditis sp. are attracted to the compound, mediated by receptors that are independent of Dop-1, leading to their death. The ß-glucosides-positive (Bgl(+)) bacteria that are otherwise non-pathogenic can obtain additional nutrients from the dead predators, thereby switching their role from prey to predator. This study also offers an evolutionary explanation for the retention by bacteria of 'cryptic' or 'silent' genetic systems such as the bgl operon.

Social determinants of diabetic retinopathy and impact of sight-threatening diabetic retinopathy: A study from Pune, India.

Purpose: Diabetes-related retinopathy is the leading cause of blindness in India. The study was carried out with the purpose of studying the association of sight-threatening diabetic retinopathy (STDR) with socioeconomic factors and demonstrating the impact of STDR on the affected individual. Methods: A mixed methods (quantitative and qualitative) research design was used. The study participants were divided into two groups for quantitative analysis. The control group consisted of non-sight-threatening diabetic retinopathy, whereas the study group consisted of sight-threatening diabetic retinopathy. Apart from demographics, data on comorbidities, type and duration of diabetes mellitus (DM), health insurance status, and socioeconomic data were collected from each individual. A statistical test (Chi-square) was performed to study the association between socioeconomic (SE) classes and STDR. For the qualitative part, a few people were chosen. Face-to-face interviews were conducted in depth. Results: A total of 207 individuals, were recruited, of which 69 had STDR and the remaining 138 had non-STDR. The incidence of STDR was high among patients with lower socioeconomic class (SEC) (upper lower and lower), and univariate analysis revealed a strong association between STDR and SEC, the presence of comorbidities, presence of health insurance, type and duration of DM, and P value <0.05. SEC, in contrast, emerged as an independent risk factor for STDR in multivariate analysis. STDR had a devastating effect on all patients interviewed. The financial impact was most likely the most severe. Conclusion: People with lower SEC are more likely to suffer from STDR-related vision loss. The impact of such vision loss on individuals is multifaceted, including a negative impact on social and work life, psychological well-being, and, most importantly, a significant financial impact.

A comparative clinical study of postoperative care by teleophthalmology and in-person consultation at hospital outpatient department after an uncomplicated cataract surgery.

BACKGROUND: Teleophthalmology provides an opportunity to conduct consultations in far-flung and remote areas that have no access to specialized eye care. However, there is a paucity of studies to assess the effect of missing in-person follow-up on initial postoperative visits. The study thus aims to compare postoperative satisfaction and uncorrected distance visual activity after an uneventful phacoemulsification cataract surgery in patients with teleconsultation approach to those with hospital visit. MATERIALS AND METHODS: The prospective observation study (n = 240) was conducted in patients who underwent surgery for cataract. Pre- and post-operative data were collected and divided into two groups based on the type of postoperative follow-up. All patients were scheduled for ophthalmic reviews in the hospital on day 1, day 7, and day 30-40 (hospital visit group) or through teleconsultation on days 1 and 7 followed by a hospital visit on days 30-40 (teleconsultation group). Outcomes evaluated in both groups were complications, patient satisfaction, and uncorrected distance visual acuity. RESULTS: Most patients in both groups were in the age group of 51-70 years. Overall satisfaction was comparable in teleconsultation and hospital visit groups (3.74 ± 0.23 vs. 3.72 ± 0.27; P = 0.22). The majority of patients had visual acuity 6/18-6/6 on postoperative day 1, day 7, and day 30-40 in both groups. Lid edema, pain, redness, watering, and congestion complications were more in the hospital visit group on postoperative day 1. CONCLUSION: The study concludes that patients with no preexisting ocular and systemic comorbidity undergoing an uneventful phacoemulsification cataract surgery teleconsultation approach can be used for follow-up without any impact on the postoperative visual outcome and patient satisfaction, thereby increasing the efficiency and productivity of health-care system.

A Novel Method to Improve Inter-Clinician Variation in the Diagnosis of Retinopathy of Prematurity Using Machine Learning.

PURPOSE: Inter-clinician variation could cause uncertainty in disease management. This is reported to be high in Retinopathy of Prematurity (ROP), a potentially blinding retinal disease affecting premature infants. Machine learning has the potential to quantify the differences in decision-making between ROP specialists and trainees and may improve the accuracy of diagnosis. METHODS: An anonymized survey of ROP images was administered to the expert(s) and the trainee(s) using a study-designed user interface. The results were analyzed for repeatability as well as to identify the level of agreement in the classification. "Ground truth" was prepared for each individual and a unique classifier was built for each individual using the same. The classifier allowed the identification of the most important features used by each individual. RESULTS: Correlation and disagreement between the expert and the trainees were visualized using the Dipstick™ diagram. Intra-clinician repeatability and reclassification statistics were assessed for all. The repeatability was 88.4% and 86.2% for two trainees and 92.1% for the expert, respectively. Commonly used features differed for the expert and the trainees and accounted for the variability. CONCLUSION: This novel, automated algorithm quantifies the differences using machine learning techniques. This will help audit the training process by objectively measuring differences between experts and trainees. TRANSLATIONAL RELEVANCE: Training for image-based ROP diagnosis can be more objectively performed using this novel, machine learning-based automated image analyzer and classifier.

Calpain4 is required for activation of HER2 in breast cancer cells exposed to trastuzumab and its suppression decreases survival and enhances response.

Overactivation of HER2 and crosstalk of other HER family members contribute to a survival pathway of breast cancer cells exposed to trastuzumab, the therapeutic inhibitor of HER2 and thus, decrease response and promote resistance. We have explored the involvement of the intracellular cysteine protease calpain4, the common partner of isoforms calpain1 and calpain2, in the regulation of cell survival and trastuzumab-response. Increase of calpain4 expression and isoform activities were detected in breast cancer cells and HER2-positive tumors. Molecular analyses of parent and resistant cells suggested that perturbation of regulations, induced by calpain4 and of activities of HER2 and HER3, was associated with trastuzumab-resistance. The suppression of calpain4 destabilized calpain1 and calpain2, however, did not prevent the activation of HER2 and HER3 or cell proliferation in the absence of trastuzumab. To understand the significance, the survival of parent and trastuzumab-resistant cells in which calpain4 was suppressed, was assessed in the presence of trastuzumab; survival in each cell type was decreased and associated with a loss of HER2 and HER3 activity. Taken together, by contributing to the activation and the crosstalk of HER2, calpain4 promotes the survival pathway of breast cancer cells, and therefore, its suppression enhances trastuzumab-response and decreases resistance.

The impact of early intervention on a child with vision loss in infancy - A parental perspective.

Purpose: : To assess the impact of early intervention services provided to children with visual loss and to report how parents perceive them in terms of a child's development and the family dynamics. Methods: : A qualitative descriptive study was conducted on a purposively selected sample of 15 children with severe visual impairment, availing early intervention services at a tertiary care facility in Pune, Maharashtra. Data were collected by conducting in-depth interviews of the parents with the help of a semi-structured interview topic guide. Participants were asked in detail about how and whether various components of the early intervention program (EIP) had an impact on their child. The interviews were audio-recorded, transcribed, and translated into English, and the resultant textual data were analyzed using the qualitative research software NVIVO 12 to identify themes and sub-themes under each domain. Results: : A total of 15 children were included in the study, with ages ranging from 13 months to 5 years. All the children included in the study suffered from severe visual impairment in infancy (Vision 3/60 - PL). In the course of this EIP, the majority of the children showed consistent progress in various aspects of child development. According to the parents, the most beneficial components of EIP were visual stimulation exercises, an improvised teaching methodology, and counseling services. Conclusion: : Almost all the parents included in the study reported a positive change in the behavior and development of the child as well as improved family dynamics after implementation of EIP.