Vision Screening Research Articles

Parental awareness on myopia prevention and control among 350 children

AIM: To understand the current situation of parental perspectives, knowledge, and practices concerning myopia prevention and control for both pre- and school-aged children. METHODS: This study was a cross-sectional survey that involved children aged 0 to 15y and their parents. Participants were required to respond to an online questionnaire by scanning a quick response (QR) code. The questionnaire consisted of 25 tick-box questions and was open to response from December 22, 2022, to January 5, 2023. The dioptric traits of the children, the visual status and educational background of the parents, the parental perspectives towards myopia and its risks, and the parents' knowledge and practices related to myopia prevention and control were recorded and measured. The Chi-square test and binomial logistic regression were used for statistics. RESULTS: Totally 350 parents responded to the questionnaire. The prevalence and severity of myopia among the surveyed children exhibited a positive correlation with advancing age (P<0.001 and P=0.004, respectively). Nearly half of parents with myopic children considered myopia did not pose any health threat and could be effectively corrected (P<0.001). Parents who held master's or doctoral degree demonstrated a better understanding of children's vision standards for each age group (P=0.001), and 31.63% of them could undergo initial vision screening for their children during the age of 0 to 3y while parents with bachelor's degree (34.04%) and below (32.43%) mainly initiated the vision examination for their children at the age of 4 to 6y (P=0.05). Parents with master's or doctoral degree also exhibited more rational practices concerning outdoor time (P=0.048) and sleep time (P=0.044). No other significant discrepancy among the different educational groups in additional conceptions of myopia, such as hyperopia reserve, axis length, and corneal curvature alterations. Most parents preferred to employ conventional interventions, such as enhancing indoor lighting condition (80.00%) and ensuring appropriate reading posture and distance (71.71%). CONCLUSION: The current status of parental knowledge and practices about myopia prevention and control remains outdated and deficient. The administrative department should implement efficacious and adaptable measures to enhance parental awareness and foster their commitment towards myopia prevention and control.

Patient-centered medical homes and vision care for children: a cross-sectional analysis with data from the national survey of children's health 2018-2019.

Vision screening as part of well-child visits is recommended annually for US children 3 to 6 years. However, 63% of children do not get a vision screening in well-child visits. The Patient-Centered Medical Home (PCMH) improves the receipt of preventive care visits in other medical specialties but it is unknown if it improves receipt of vision screening. The objective of this study is to determine whether caregiver-reported receipt of care in a PCMH is associated with receiving a vision screening test in a well-child visit for children 3 to 6 years in a pediatrician/general doctor's office ('primary care'). Population-based data for US children aged 3 to 6 years was derived from the National Survey of Children's Health (2018-2019). Children were excluded if they did not have a well-child visit in the previous 12 months. The primary exposure was receipt of care in a PCMH ('PCMH care'), and the primary outcome was receipt of a vision screening in primary care. Adjusted odds ratios (aOR) and predicted probabilities were computed for children with and without PCMH care. Among 9,587 children with well-child visits, 4,984 (50.9%) were males. There were 1,107 (23.3%) Hispanic, 6704 (52.8%) White/Non-Hispanic and 545 (11.8%) Black/Non-Hispanic children. There were 5,482 (51.8%) children who received PCMH care. Of those with PCMH care, 2,629 (52.2%) received a vision screening in primary care. Those with PCMH care (52.2%) had a higher odds of receiving a vision screening (aOR, 1.31; 95% CI 1.11, 1.55) in primary care compared to children without PCMH care (42.8%). The predicted probability of vision screening in primary care was 50.8% (95% CI 48.2, 53.5) for children that received PCMH care, and 44.3% (95% CI 41.3, 47.3) without PCMH care. Caregiver-reported receipt of PCMH care was associated with a greater likelihood of vision screening for children 3 to 6 years in a well-child visit. Further studies are needed to understand the mechanisms through which PCMH care contributes to the greater odds of vision screening in primary care to inform the creation of strategic interventions to prevent vision loss and its long-term implications.

Sirtuin inhibitors reduce intracellular growth of M. tuberculosis in human macrophages via modulation of host cell immunity.

Host-directed therapies aiming to strengthen the body's immune system, represent an underexplored opportunity to improve treatment of tuberculosis (TB). We have previously shown in Mycobacterium tuberculosis (Mtb)-infection models and clinical trials that treatment with the histone deacetylase (HDAC) inhibitor, phenylbutyrate (PBA), can restore Mtb-induced impairment of antimicrobial responses and improve clinical outcomes in pulmonary TB. In this study, we evaluated the efficacy of different groups of HDAC inhibitors to reduce Mtb growth in human immune cells. A panel of 21 selected HDAC inhibitors with different specificities that are known to modulate infection or inflammation was tested using high-content live-cell imaging and analysis. Monocyte-derived macrophages or bulk peripheral blood cells (PBMCs) were infected with the green fluorescent protein (GFP)-expressing Mtb strains H37Ra or H37Rv and treated with HDAC inhibitors in the micromolar range in parallel with a combination of the first-line antibiotics, rifampicin, and isoniazid. Host cell viability in HDAC inhibitor treated cell cultures was monitored with Cytotox-red. Seven HDAC inhibitors were identified that reduced Mtb growth in macrophages > 45-75% compared to average 40% for PBA. The most effective compounds were inhibitors of the class III HDAC proteins, the sirtuins. While these compounds may exhibit their effects by improving macrophage function, one of the sirtuin inhibitors, tenovin, was also highly effective in extracellular killing of Mtb bacilli. Antimicrobial synergy testing using checkerboard assays revealed additive effects between selected sirtuin inhibitors and subinhibitory concentrations of rifampicin or isoniazid. A customized macrophage RNA array including 23 genes associated with cytokines, chemokines and inflammation, suggested that Mtb-infected macrophages are differentially modulated by the sirtuin inhibitors as compared to PBA. Altogether, these results demonstrated that sirtuin inhibitors may be further explored as promising host-directed compounds to support immune functions and reduce intracellular growth of Mtb in human cells.

Cytologic hallmarks and differential diagnosis of papillary thyroid carcinoma subtypes

Papillary thyroid carcinoma (PTC) is the most common thyroid malignancy, characterized by a range of subtypes that differ in their cytologic features, clinical behavior, and prognosis. Accurate cytologic evaluation of PTC using fine-needle aspiration is essential but can be challenging due to the morphologic diversity among subtypes. This review focuses on the distinct cytologic characteristics of various PTC subtypes, including the classic type, follicular variant, tall cell, columnar cell, hobnail, diffuse sclerosing, Warthin-like, solid/trabecular, and oncocytic PTCs. Each subtype demonstrates unique nuclear features, architectural patterns, and background elements essential for diagnosis and differentiation from other thyroid lesions. Recognizing these distinct cytologic patterns is essential for identifying aggressive subtypes like tall cell, hobnail, and columnar cell PTCs, which have a higher risk of recurrence, metastasis, and poorer clinical outcomes. Additionally, rare subtypes such as diffuse sclerosing and Warthin-like PTCs present unique cytologic profiles that must be carefully interpreted to avoid diagnostic errors. The review also highlights the cytologic indicators of lymph node metastasis and high-grade features, such as differentiated high-grade thyroid carcinoma. The integration of molecular testing can further refine subtype diagnosis by identifying specific genetic mutations. A thorough understanding of these subtype-specific cytologic features and molecular profiles is vital for accurate diagnosis, risk stratification, and personalized management of PTC patients. Future improvements in diagnostic techniques and standardization are needed to enhance cytologic evaluation and clinical decision-making in thyroid cancer.

Benzocarbazoledinones as SARS-CoV-2 Replication Inhibitors: Synthesis, Cell-Based Studies, Enzyme Inhibition, Molecular Modeling, and Pharmacokinetics Insights

Endemic and pandemic viruses represent significant public health challenges, leading to substantial morbidity and mortality over time. The COVID-19 pandemic has underscored the urgent need for the development and discovery of new, potent antiviral agents. In this study, we present the synthesis and anti-SARS-CoV-2 activity of a series of benzocarbazoledinones, assessed using cell-based screening assays. Our results indicate that four compounds (4a, 4b, 4d, and 4i) exhibit EC50 values below 4 μM without cytotoxic effects in Calu-3 cells. Mechanistic investigations focused on the inhibition of the SARS-CoV-2 main protease (Mpro) and papain-like protease (PLpro) have used enzymatic assays. Notably, compounds 4a and 4b showed Mpro inhibition activity with IC50 values of 0.11 ± 0.05 and 0.37 ± 0.05 µM, respectively. Furthermore, in silico molecular docking, physicochemical, and pharmacokinetic studies were conducted to validate the mechanism and assess bioavailability. Compound 4a was selected for preliminary drug-likeness analysis and in vivo pharmacokinetics investigations, which yielded promising results and corroborated the in vitro and in silico findings, reinforcing its potential as an anti-SARS-CoV-2 lead compound.

Nicotinamide and pyridoxine stimulate muscle stem cell expansion and enhance regenerative capacity during aging.

Skeletal muscle relies on resident muscle stem cells (MuSCs) for growth and repair. Aging and muscle diseases impair MuSC function, leading to stem cell exhaustion and regenerative decline that contribute to the progressive loss of skeletal muscle mass and strength. In the absence of clinically available nutritional solutions specifically targeting MuSCs, we used a human myogenic progenitor (hMP) high-content imaging screen of natural molecules from food to identify nicotinamide (NAM) and pyridoxine (PN) as bioactive nutrients that stimulate MuSCs and have history of safe human use. NAM and PN synergize via CK1-mediated cytoplasmic β-catenin activation and AKT signaling to promote amplification and differentiation of MuSCs. Oral treatment with a combination of NAM/PN accelerates muscle regeneration in vivo by stimulating MuSCs, increases muscle strength during recovery, and overcomes MuSC dysfunction and regenerative failure during aging. Levels of NAM and bioactive PN spontaneously decline during aging in model organisms and inter-independently associate with muscle mass and walking speed in a human cohort of 186 aged people. Collectively, our results establish NAM/PN as a new nutritional intervention that stimulates MuSCs, enhances muscle regeneration, and alleviates age-related muscle decline with a direct opportunity for clinical translation.

An automated high-content screening and assay platform for the analysis of spheroids at subcellular resolution.

The endomembrane system is essential for healthy cell function, with the various compartments carrying out a large number of specific biochemical reactions. To date, almost all of our understanding of the endomembrane system has come from the study of cultured cells growing as monolayers. However, monolayer-grown cells only poorly represent the environment encountered by cells in the human body. As a first step to address this disparity, we have developed a platform that allows us to investigate and quantify changes to the endomembrane system in three-dimensional (3D) cell models, in an automated and highly systematic manner. HeLa Kyoto cells were grown on custom-designed micropatterned 96-well plates to facilitate spheroid assembly in the form of highly uniform arrays. Fully automated high-content confocal imaging and analysis were then carried out, allowing us to measure various spheroid-, cellular- and subcellular-level parameters relating to size and morphology. Using two drugs known to perturb endomembrane function, we demonstrate that cell-based assays can be carried out in these spheroids, and that changes to the Golgi apparatus and endosomes can be quantified from individual cells within the spheroids. We also show that image texture measurements are useful tools to discriminate cellular phenotypes. The automated platform that we show here has the potential to be scaled up, thereby allowing large-scale robust screening to be carried out in 3D cell models.

A Cell-Based Screening Assay for rRNA-Targeted Drug Discovery.

Worldwide, bacterial antibiotic resistance continues to outpace the level of drug development. One way to counteract this threat to society is to identify novel ways to rapidly screen and identify drug candidates in living cells. Developing fluorescent antibiotics that can enter microorganisms and be displaced by potential antimicrobial compounds is an important but challenging endeavor due to the difficulty in entering bacterial cells. We developed a cell-based assay using a fluorescent aminoglycoside molecule that allows for the rapid and direct characterization of aminoglycoside binding in a population of bacterial cells. The assay involves the accumulation and competitive displacement of a fluorescent aminoglycoside binding probe in Escherichia coli as a Gram-negative bacterial model. The assay was optimized for high signal-to-background ratios, ease of performance for reliable outcomes, and amenability to high-throughput screening. We demonstrate that the fluorescent binding probe shows a decrease in fluorescence with cellular uptake, consistent with RNA binding, and also shows a subsequent increase upon the addition of the positive control neomycin. Fluorescence intensity increase with aminoglycosides was indicative of their relative binding affinities for A-site rRNA, with neomycin having the highest affinity, followed by paromomycin, tobramycin, sisomicin, and netilmicin. Intermediate fluorescence was found with plazomicin, neamine, apramycin, ribostamicin, gentamicin, and amikacin. Weak fluorescence was observed with kanamycin, hygromycin, streptomycin, and spectinomycin. A high degree of sensitivity was observed with aminoglycosides known to be strong binders for the 16S rRNA A-site compared with antibiotics that target other biosynthetic pathways. The quality of the optimized assay was excellent for planktonic cells, with an average Z' factor value of 0.80. In contrast to planktonic cells, established biofilms yielded an average Z' factor of 0.61. The high sensitivity of this cell-based assay in a physiological context demonstrates significant potential for identifying potent new ribosomal binding antibiotics.

Bilateral Macular Dysplasia in Coffin-Siris Syndrome.

To report a case of bilateral macular dysplasia without pigmentary retinopathy in a patient with genetically confirmed Coffin-Siris syndrome (CSS) and a co-existing, heterozygous CNGB1 variant. Case report. A 7-year-old boy who failed his vision screening was referred for retinal consultation after his eye care provider identified bilateral macular deposits. Past medical history included developmental delay, atypical facial features, uteropelvic junction obstruction, koilonychia, and corpus callosum dysgenesis. The patient underwent whole exome sequencing, which revealed a pathogenic variant in the ARID1A gene consistent with a diagnosis of Coffin-Siris syndrome. Genetic testing also revealed a heterozygous, missense variant in the CNGB1 gene of unknown significance. Fundus examination revealed fine hypopigmented lesions in the macula with surrounding retinal pigment epithelial changes in both eyes. Spectral-domain ocular coherence tomography revealed abnormalities of the ellipsoid and interdigitation zones. The patient's best-corrected visual acuity and clinical examination remained stable for a decade since his initial examination, with persistent macular dysplasia and no pigmentary retinopathy. It is possible that the ocular findings in this case may expand the phenotypes associated with CSS.

Refractive Profiles in Children Receiving School-Based Eye Exams Following Vision Screening from a Large School-Based Vision Program in 2016 to 2022

ABSTRACT Purpose To describe refractive error findings and associated factors in students who received school-based eye exams following vision screenings. Methods Cross-sectional study of pre-kindergarten through 12th grade students who failed vision screening and underwent a school-based eye exam in the Northeast region of the United States during 2016–2022. Non-cycloplegic autorefraction and visual acuity measurements were used to categorize refractive error by type and severity. Main outcomes included any refractive error (at least −0.50D myopia, +0.50 hyperopia, 1.00D astigmatism, or 1.00D anisometropia), clinically significant refractive error (CSRE; more severe refractive error with decreased vision), and refractive amblyopia risk (RAR). Multivariable mixed-effects logistic regression was used to determine student- and school-level characteristics associated with refractive error outcomes. Results Of 103,159 included students who failed screening, 95,875 (92.9%) were analyzed. Overall prevalence of any refractive error was 94.2%, with 81.7% having CSRE. Pre-kindergarten & kindergarten students had the highest prevalence of CSRE (85.7%), which dropped to 77.0% by 3rd and 4th grade before rising with each higher grade level thereafter. Prevalence of RAR was 60.9% overall and highest in pre-kindergarten and kindergarten students (73.8%). Myopia was the most prevalent refractive error, followed by astigmatism, anisometropia, then hyperopia. The odds of hyperopia, astigmatism, and anisometropia decreased with higher grade level. Conclusion Over 80% of students who failed vision screening at a large school-based vision program had CSRE, and over 60% had RAR. Pre-kindergarten and kindergarten students had an especially high prevalence of RAR. Refractive error remained common in every grade level.

Evaluation of the diagnostic parameters of the amblyopia and risk factors for amblyopia screening protocol in 3-year-olds according to recommendations from the French Association for Pediatric Ophthalmology and Strabismus (AFSOP) compared with reference ophthalmological examination: the ORTHOPHTALMO study

ABSTRACT Introduction: The ORTHOPHTALMO study aims to evaluate the diagnostic parameters of the screening protocol for amblyopia and risk factors for amblyopia in 3-year-olds recommended in 2019 by the French Association for Pediatric Ophthalmology and Strabismus (AFSOP). This protocol uses visual acuity, photoscreening refraction, and cover test examination performed by an orthoptist. Patient referral to an ophthalmologist is only according to recommended referral criteria. Methods: A prospective, single-center study was performed between September 2020 and June 2021 on a consecutive series of 3-year-olds consulting the Ophthalmology Centre of Clinique Rive Gauche, Toulouse, France, for vision screening. Patients were first examined by an orthoptist following the screening protocol recommended by AFSOP. All patients were then systematically examined by an ophthalmologist for cycloplegic refraction measurement (reference examination). The ophthalmologist was blinded to the referral conclusion and refraction measurements of the orthoptist. Results: A total of 300 patients (149 girls and 151 boys) were included. Examination by an orthoptist was unreliable/incomplete in 7% of cases. An abnormality was detected by the orthoptist in ≥1 of the screening tests among 42% of patients; these patients were thus considered as requiring referral to an ophthalmologist. Reference ophthalmological examination found 41% of patients required treatment. The diagnostic parameters of this screening protocol were 90% for sensitivity and 89% for specificity. Discussion: We validate the effectiveness and feasibility of the AFSOP screening protocol for detection of amblyopia and risk factors for amblyopia in 3-year-olds as well as the recommended criteria for referral to an ophthalmologist. ClinicalTrials.gov https://clinicaltrials.gov/ct2/show/NCT04395560. Number: NCT04395560.

Vision Screening and Detection of Ocular Abnormalities in School Children by Teachers in Jordan.

To evaluate the accuracy of vision screening and detection of ocular abnormalities conducted by teachers in school children in Jordan compared with vision testing by optometrists and ocular disease identification by ophthalmologists. A non-random, purposive sampling strategy where 6-year-old and 15-year-old school children from three schools in Amman, Jordan were included. Twenty-two teachers were trained to conduct visual acuity screening using the 0.2 logMAR line of the distance visual acuity (DVA) chart at 10 feet and near visual acuity (NVA) chart at 16 inches, in addition to detecting ocular abnormalities. An optometrist assessed the visual acuity and conducted objective and subjective refraction, while an ophthalmologist examined the ocular health of all children. A total of 542 children (51% female) were included in the study, of which 47% were 6 years old and 53% were 15 years old. Teacher screening had sensitivities of 76.92% for DVA, 68% for NVA, and 37.50% for abnormality detection. The specificities of DVA, NVA, and ocular abnormalities were 98.22%, 98.48%, and 99.24%, respectively. The positive predictive value for DVA, NVA, and ocular abnormalities were 83.33%, 99.00%, and 60%, respectively. The negative predictive value for each procedure was 98.22% for DVA, 98.48% for NVA, and 98.12% for ocular abnormalities. Teachers were able to conduct vision screening with a high level of accuracy compared to the gold standard of testing by optometrists, which would be useful for the early detection and referral of refractive errors in school children. However, they were unable to detect ocular abnormalities compared with the gold standard for disease identification by ophthalmologists. Further training and monitoring, or different training approaches should be implemented to enable teachers to identify ocular abnormalities at acceptable levels.

Lost in translation: Illuminating protein mislocalization through high-content screening microscopy

Establishing the subcellular distribution of all proteins encoded by the human genome remains a key objective of life science research. This is particularly important in the context of proteins that, through genetic sequencing of patients, have been identified as containing missense mutations. A recent publication in Cell1 highlights the prominence of protein mislocalization as a hallmark of dysfunctional proteins. The use of high-content subcellular phenotypic screens and allied technology by Lacoste and colleagues has enormous potential to change the landscape of how we approach both diagnostic and therapeutic decisions.

Comparison of the GoCheck Kids and Spot Screener photoscreening devices for the detection of amblyopia risk factors using 2021 AAPOS recommendations

PurposeTo compare the Spot Vision Screener (SPOT) to the GoCheck Kids mobile application (GCK) in the evaluation of amblyopia risk factors (ARFs), according to updated 2021 AAPOS guidelines for instrument-based pediatric vision screening. MethodsSPOT, GCK using horizontal and vertical images (two-photo mode), and a complete eye examination (CEE) were performed on children ages ≥1 to <7 years referred to a pediatric ophthalmologist from October 2020 to December 2022 for a failed vision screen based on manufacturer guidelines. The primary outcomes were sensitivity, specificity, and positive predictive value (PPV) for each photoscreener. ResultsOf 267 subjects included, 42% had one or more 2021 guideline ARFs after CEE. Overall, for SPOT and GCK, respectively, the sensitivity was 0.765 and 0.748, specificity was 0.769 and 0.755, and PPV was 0.728 and 0.712 (P > 0.05 for all). In participants ≥4 years old, SPOT had higher ARF sensitivity than GCK (0.900 and 0.750, resp.; P = 0.005), while among participants <4 years old, SPOT had lower sensitivity than GCK (0.487 and 0.744 respectively; P = 0.004). There were no statistically significant differences between SPOT and GCK in specificity among the younger (0.750 and 0.676 resp.) or older (0.785 and 0.823, resp.) cohorts, or in strabismus detection. The performance of the SPOT and GCK were evaluated using receiver operating characteristics (ROC) curves. Area under the curve (AUC) ranged from 0.857 to 0.980 for SPOT and 0.809-0.931 for GCK. ConclusionsOverall, SPOT, and GCK had similar values for sensitivity, specificity, and PPV to identify patients with ARFs using the 2021 AAPOS guidelines.

Associations between School-Based Vision Program Outcomes and School Characteristics in 410 Schools.

School-based vision programs (SBVPs) deliver vision care services directly to students at school, helping address disparities in access to pediatric vision care. We aim to evaluate the associations between SBVP outcomes and school-level characteristics. Retrospective cross-sectional data analysis PARTICIPANTS: Public schools with at least 50 SBVP-enrolled students 5 to 22 years old with complete demographic data. Schools with fewer than 60% of total grade levels served by the SBVP were excluded, resulting in a sample of 410 schools. Vision screening and eye exam data were extracted from Helen Keller International's United States Vision Program dataset from 2016 through 2022. Individual student data were aggregated to characterize each school's SBVP outcomes and analyzed with schools' publicly available socioeconomic and demographic data (student body race and ethnicity composition, proportion of students qualifying for free and reduced-price meals [FARM%], and proportion of English Language Learners). Descriptive statistics summarized SBVP outcomes. Fractional regression models were utilized to understand associations between SBVP outcomes and school characteristics. SBVP outcomes were rates of vision screening failure, eyeglasses prescription, and community eye care referral among each school's SBVP-enrolled students. We evaluated 151 (36.8%) elementary, 155 (37.8%) middle, and 104 (25.4%) high schools, with a median FARM% of 87.4% and a plurality of Hispanic students in 61.0% of schools. Overall median rates of vision screening failure, eyeglasses prescription, and referral were 38.4%, 25.2%, and 5.4%, respectively. High schools were associated with increased screening failure and eyeglasses prescription rates and a decrease in referral rate, compared with elementary schools. In multivariate analysis, each 10% increase in FARM% was associated with a 2.6% (95% confidence interval [CI]: 1.54-3.65), 1.8% (CI: 0.87-2.74), and 0.86% (CI: 0.36-1.36) increase in screening failure, eyeglasses prescription, and referral rates, respectively. There is significant vision care demand among public schools, especially those with student populations from lower socioeconomic backgrounds. SBVPs are important in bridging the gap in pediatric vision care access in such populations. Our findings demonstrated opportunities to allocate program staffing and equipment resources according to schools' anticipated needs, thus maximizing SBVPs' impact in delivering pediatric vision care.

Potential treatment approaches for malignant peritoneal mesothelioma: in vivo and in vitro experimental study of natural killer cell immunotherapy.

Malignant peritoneal mesothelioma (MPM) is a rare primary malignant tumor with an extremely poor prognosis that currently lacks effective treatment options. This study investigated the in vitro and in vivo efficacy of natural killer (NK) cells for treatment of MPM. An in vitro study was conducted to assess the cytotoxicity of NK cells from umbilical cord blood to MPM cells with the use of a high-content imaging analysis system, the Cell Counting Kit-8 assay, and Wright-Giemsa staining. The level of NK cell effector molecule expression was detected by flow cytometry and enzyme-linked immunosorbent assays. The ability of NK cells to kill MPM cells was determined based on live cell imaging, transmission electron microscopy, and scanning electron microscopy. An in vivo study was conducted to assess the efficacy and safety of NK cell therapy based on the experimental peritoneal cancer index, small animal magnetic resonance imaging, and conventional histopathologic, cytologic, and hematologic studies. NK cells effectively killed MPM cells through the release of effector molecules (granzyme B, perforin, interferon-γ, and tumor necrosis factor-α) in a dose- and density-dependent manner. The NK cell killing process potentially involved four dynamic steps: chemotaxis; hitting; adhesion; and penetration. NK cells significantly reduced the tumor burden, diminished ascites production, and extended survival with no significant hematologic toxicity or organ damage in NOG mice. NK cell immunotherapy inhibited proliferation of MPM cells in vitro and in vivo with a good safety profile.

Cladophorol-A is an inhibitor of cyclic GMP-AMP synthase

Cyclic guanosine monophosphate (GMP)-adenosine monophosphate (AMP) synthase (cGAS) is an enzyme sensor of double-stranded DNA (dsDNA) that serves to trigger activation of the cGAS-stimulator of interferon genes (STING) pathway. Excessive activation of this pathway has been demonstrated to contribute to various forms of inflammatory disease. As such, cGAS has arisen as a potential therapeutic target with broad potential applications. Using a pathway-targeted cell-based screening approach, we identified the natural product Cladophorol A as a new class of non-cytotoxic cGAS inhibitor (cell-based IC50 = 370 nM). An x-ray co-crystal structure at 2.75 Å resolution revealed that Cladophorol A inhibits cGAS by binding to its active site within the conserved adenosine nucleobase binding site.

Vision screening for preschoolers with commercial insurance: impact of geography

Vision screening for preschoolers with commercial insurance: impact of geography

An image-based RNAi screen identifies the EGF.R signaling pathway as a regulator of Imp/ IGF2BP RNP granules.

Biomolecular condensates have recently retained much attention since they provide a fundamental mechanism of cellular organization. Among those, cytoplasmic RNP granules selectively and reversibly concentrate RNA molecules and regulatory proteins, thus contributing to the spatio-temporal regulation of associated RNAs. Extensive in vitro work has unraveled the molecular and chemical bases of RNP granule assembly. The signaling pathways controlling this process in a cellular context are however still largely unknown. Here, we aimed at identifying regulators of cytoplasmic RNP granules characterized by the presence of the evolutionarily conserved IGF2BP/Imp/ZBP1 RNA binding protein. We performed a high-content image-based RNAi screen targeting all Drosophila genes encoding RNA binding proteins, phosphatases and kinases. This led to the identification of dozens of genes regulating the number of Imp+ RNP granules in S2R+ cells, among which components of the MAPK pathway. Combining functional approaches, phospho-mapping and generation of phospho-variants, we further showed that the EGF.R signaling inhibits Imp+ RNP granule assembly through activation of MAPK/Rolled and Imp S15 phosphosite. This work illustrates how signaling pathways can regulate cellular condensate assembly by post-translational modifications of specific components.

Seq-Scope: repurposing Illumina sequencing flow cells for high-resolution spatial transcriptomics.

Spatial transcriptomics technologies aim to advance gene expression studies by profiling the entire transcriptome with intact spatial information from a single histological slide. However, the application of spatial transcriptomics is limited by low resolution, limited transcript coverage, complex procedures, poor scalability and high costs of initial setup and/or individual experiments. Seq-Scope repurposes the Illumina sequencing platform for high-resolution, high-content spatial transcriptome analysis, overcoming these limitations. It offers submicrometer resolution, high capture efficiency, rapid turnaround time and precise annotation of histopathology at a much lower cost than commercial alternatives. This protocol details the implementation of Seq-Scope with an Illumina NovaSeq 6000 sequencing flow cell, allowing the profiling of multiple tissue sections in an area of 7 mm × 7 mm or larger. We describe the preparation of a fresh-frozen tissue section for both histological imaging and sequencing library preparation and provide a streamlined computational pipeline with comprehensive instructions to integrate histological and transcriptomic data for high-resolution spatial analysis. This includes the use of conventional software tools for single-cell and spatial analysis, as well as our recently developed segmentation-free method for analyzing spatial data at submicrometer resolution. Aside from array production and sequencing, which can be done in batches, tissue processing, library preparation and running the computational pipeline can be completed within 3 days by researchers with experience in molecular biology, histology and basic Unix skills. Given its adaptability across various biological tissues, Seq-Scope establishes itself as an invaluable tool for researchers in molecular biology and histology.