Highly efficient capture approach for the identification of diverse inherited retinal disorders.

Our study presents a 319-gene panel targeting inherited retinal dystrophy (IRD) genes. Through a multi-center retrospective cohort study, we validated the assay's effectiveness and clinical utility and characterized the mutation spectrum of Taiwanese IRD patients. Between January 2018 and May 2022, 493 patients in 425 unrelated families, all initially suspected of having IRD without prior genetic diagnoses, underwent detailed ophthalmic and physical examinations (with extra-ocular features recorded) and genetic testing with our customized panel. Disease-causing variants were identified by segregation analysis and clinical interpretation, with validation via Sanger sequencing. We achieved a read depth of >200× for 94.2% of the targeted 1.2 Mb region. 68.5% (291/425) of the probands received molecular diagnoses, with 53.9% (229/425) resolved cases. Retinitis pigmentosa (RP) is the most prevalent initial clinical impression (64.2%), and 90.8% of the cohort have the five most prevalent phenotypes (RP, cone-rod syndrome, Usher's syndrome, Leber's congenital amaurosis, Bietti crystalline dystrophy). The most commonly mutated genes of probands that received molecular diagnosis are USH2A (13.7% of the cohort), EYS (11.3%), CYP4V2 (4.8%), ABCA4 (4.5%), RPGR (3.4%), and RP1 (3.1%), collectively accounted for 40.8% of diagnoses. We identify 87 unique unreported variants previously not associated with IRD and refine clinical diagnoses for 21 patients (7.22% of positive cases). We developed a customized gene panel and tested it on the largest Taiwanese cohort, showing that it provides excellent coverage for diverse IRD phenotypes.

Microglia Reduce Herpes Simplex Virus 1 Lethality of Mice with Decreased T Cell and Interferon Responses in Brains.

Herpes simplex virus 1 (HSV-1) infects the majority of the human population and can induce encephalitis, which is the most common cause of sporadic, fatal encephalitis. An increase of microglia is detected in the brains of encephalitis patients. The issues regarding whether and how microglia protect the host and neurons from HSV-1 infection remain elusive. Using a murine infection model, we showed that HSV-1 infection on corneas increased the number of microglia to outnumber those of infiltrating leukocytes (macrophages, neutrophils, and T cells) and enhanced microglia activation in brains. HSV-1 antigens were detected in brain neurons, which were surrounded by microglia. Microglia depletion increased HSV-1 lethality of mice with elevated brain levels of viral loads, infected neurons, neuron loss, CD4 T cells, CD8 T cells, neutrophils, interferon (IFN)-ß, and IFN-γ. In vitro studies demonstrated that microglia from infected mice reduced virus infectivity. Moreover, microglia induced IFN-ß and the signaling pathway of signal transducer and activator of transcription (STAT) 1 to inhibit viral replication and damage of neurons. Our study reveals how microglia protect the host and neurons from HSV-1 infection.

Doxycycline Ameliorates the Severity of Experimental Proliferative Vitreoretinopathy in Mice.

After successful surgeries for patients with rhegmatogenous retinal detachment, the most common cause of retinal redetachment is proliferative vitreoretinopathy (PVR), which causes severe vision impairment and even blindness worldwide. Until now, the major treatment for PVR is surgical removal of the epiretinal membrane, while effective treatment to prevent PVR is still unavailable. Therefore, we investigated the potential of doxycycline, an antibiotic in the tetracycline class, to treat PVR using a mouse model. We used the human retinal pigment epithelial cell line, ARPE-19, for in vitro and in vivo studies to test doxycycline for PVR treatment. We found that doxycycline suppressed the migration, proliferation, and contraction of ARPE-19 cells with reduced p38 MAPK activation and total MMP activity. Intravitreal doxycycline and topical tetracycline treatment significantly ameliorated the PVR severity induced by ARPE-19 cells in mice. PVR increased the expression of MMP-9 and IL-4 and p38 MAPK phosphorylation and modestly decreased IL-10. These effects were reversed by doxycycline and tetracycline treatment in the mouse retina. These results suggest that doxycycline will be a potential treatment for PVR in the future.

Chitosan Oligosaccharides Suppress Nuclear Factor-Kappa B Activation and Ameliorate Experimental Autoimmune Uveoretinitis in Mice.

We investigated the therapeutic potential and mechanism of chitosan oligosaccharides (COS) for experimental autoimmune uveoretinitis (EAU) in mice. EAU was induced in C57/BL6 mice by injection of human interphotoreceptor retinoid-binding protein (IRBP) peptides. At the same time, a high or low dose (20 or 10 mg/kg) of COS or phosphate-buffered saline (PBS) was given to mice daily after EAU induction. We found that mouse EAU is ameliorated by the high-dose COS treatment when compared with PBS treatment. In the retinas of high-dose COS-treated mice, the nuclear translocation of NF-κB subunit (p65) was suppressed, and the expression of several key EAU inflammatory mediators, IFN-γ, TNF-α, IL-1α, IL-4, IL-5, IL-6, IL-10, IL-17 and MCP-1 was lowered. These results suggest that COS may be a potential treatment for posterior uveitis.

Suppression of the Reactive Oxygen Response Alleviates Experimental Autoimmune Uveitis in Mice.

Reactive oxygen species (ROS) are produced by host phagocytes and play an important role in antimicrobial actions against various pathogens. Autoimmune uveitis causes blindness and severe visual impairment in humans at all ages worldwide. However, the role of ROS in autoimmune uveitis remains unclear. We used ROS-deficient (Ncf1-/-) mice to investigate the role of ROS in experimental autoimmune uveitis (EAU). Besides, we also used the antioxidant N-acetylcysteine (NAC) treatment to evaluate the effect of suppression of ROS on EAU in mice. The EAU disease scores of Ncf1-/- mice were significantly lower than those of wild-type mice. EAU induction increased the levels of cytokines (interleukin (IL)-1α, IL-1ß, IL-4, IL-6, IL-12, IL-17, and tumor necrosis factor (TNF)-α) and chemokines (monocyte chemoattractant protein (MCP)-1) in the retinas of wild-type mice but not in those of Ncf1-/- mice. EAU induction enhanced the level of NF-κB activity in wild-type mice. However, the level of NF-κB activity in Ncf1-/- mice with EAU induction was low. Treatment with the antioxidant NAC also decreased the severity of EAU in mice with reduced levels of oxidative stress, inflammatory mediators, and NF-κB activation in the retina. We successfully revealed a novel role of ROS in the pathogenesis of EAU and suggest a potential antioxidant role for the treatment of autoimmune uveitis in the future.

Bortezomib induces HSV-1 lethality in mice with neutrophil deficiency.

Bortezomib suppressing NF-κB activity is an effective therapy for patients with myeloma or lymphoma. However, this drug can cause adverse effects, neutropenia, and recurrent infections of herpes viruses. Among herpes viruses, HSV-1 can reactivate to induce mortality. The important issues regarding how bortezomib diminishes neutrophils, whether bortezomib can induce HSV-1 reactivation, and how bortezomib exacerbates HSV-1 infection, need investigation. Using the murine model, we found that bortezomib induced HSV-1 reactivation. Bortezomib diminished neutrophil numbers in organs of uninfected and HSV-1-infected mice and turned a nonlethal infection to lethal with elevated tissue viral loads. In vitro results showed that bortezomib and HSV-1 collaborated to enhance the death and apoptosis of mouse neutrophils. The leukocyte deficiency induced by chemotherapies is generally believed to be the cause for aggravating virus infections. Here we show the potential of pathogen to exacerbate chemotherapy-induced leukocyte deficiency.

The influence of orthokeratology on intraocular pressure measurements.

PURPOSE: Underestimation of IOP in a myopic patient may lead to misjudgment of the risk of glaucoma. This study investigated the influence of orthokeratology-induced change in CCT on IOP measured by the non-contact pneumotonometer (NCT), Goldmann applanation tonometer (GAT), and Pascal dynamic contour tonometer (PDCT). METHODS: This study was conducted to examine the eyes of 34 patients who received orthokeratology for myopia. CCT and IOP were measured, and IOP was obtained with the NCT, GAT, and PDCT. The associations between changes in measured IOP and change in CCT at different orthokeratology follow-up time points were evaluated by linear regression analysis. RESULTS: Change in IOP measured by the three tonometries correlated significantly with change in CCT after one-week application of orthokeratology. The correlation was strongest for NCT followed by GAT and PDCT. The changes in measured IOP corresponding to a 10 µm decrease in CCT were 0.7-0.9, 0.4-0.6, and 0.2-0.3 mm Hg for NCT, GAT, and PDCT, respectively. CONCLUSIONS: The IOP measured by the three methods--NCT, GAT, and PDCT--decreased as a result of the change in CCT induced by orthokeratology. The influence on NCT and GAT was greater than that on PDCT.

Relationship between air pollution and outpatient visits for nonspecific conjunctivitis.

PURPOSE: Past studies present evidence of associations between air pollution and human ocular symptoms; however, to the knowledge of the authors, research investigating the hazardous effects of air pollution on nonspecific conjunctivitis is nonexistent. This study investigates the relationship between air pollution and outpatient visits for nonspecific conjunctivitis in Taiwan. A multiarea analysis was conducted to examine and assess the risks of short-term effects of particulate matter (PM), nitrogen dioxide (NO2), sulfur dioxide (SO2), ozone (O3), and carbon monoxide on nonspecific conjunctivitis. METHODS: Data were collected from outpatient visits for nonspecific conjunctivitis from seven air-quality-monitoring areas. To find immediate and lag effects of air pollution, an area-specific, case-crossover analysis was performed and a meta-analysis with random effects was used to combine the area-specific RESULTS: Results. The effects on outpatient visits for nonspecific conjunctivitis are strongest for O3 and NO2, with a 2.5% increase (95% confidence interval [CI], 0.9-4.1) for a 16.4 ppb (parts per billion) concentration rise in O3 and a 2.3% increase (95% CI, 0.7-3.9) for an 11.47 ppb concentration rise in NO2. Effects are also found for particulate matter with an aerodynamic diameter ≤ 10 µm (PM10) and SO2. Effects are more prominent in winter because the analysis was stratified according to season. CONCLUSIONS: The air pollutants NO2, SO2, O3, and PM10 increase the chances of outpatient visits for nonspecific conjunctivitis and have no evident lag effects.

Endogenous Klebsiella endophthalmitis associated with pyogenic liver abscess.

OBJECTIVE: To study risk factors, clinical features, treatment, and visual outcomes in patients with endogenous Klebsiella pneumoniae endophthalmitis (EKE) associated with K. pneumoniae-induced pyogenic liver abscess, and to investigate contributing factors in successfully treated cases. DESIGN: Retrospective, noncomparative, interventional case series. PARTICIPANTS: Review of medical records of 22 consecutive patients with EKE and pyogenic liver abscess. METHODS: The affected eyes of 22 consecutive patients (n = 27) with EKE, who presented to our ophthalmic service during a recent 8-year period, were studied retrospectively. MAIN OUTCOME MEASURES: Best-corrected visual acuity (VA) at end of follow up. RESULTS: Diabetes mellitus was the most common comorbid risk factor (n = 15 [68%]). Five patients (23%) had bilateral eye involvement. On initial presentation, characteristic pupillary hypopyon was observed in 12 eyes. Diagnosis was confirmed by blood culture in 8 patients, culture of liver aspirate in 17 patients, and vitreous culture in 11 patients. Other associated septic metastatic lesions included pulmonary abscess or emboli in 6 cases, brain abscess or meningitis in 3 cases, and prostate and kidney abscesses in 1 case. Despite aggressive intravenous and intravitreal antibiotic therapy, final VA of light perception or worse affected 24 eyes (89%), of which 11 (41%) were eventually eviscerated or enucleated. Successful treatment with retained useful vision better than 6/60 was achieved in 3 eyes, of which 2 received early intravitreal corticosteroid injections. However, the other remaining eye had a focal subretinal abscess. CONCLUSION: Physicians should be alert to the development of EKE when patients with diabetes along with K. pneumoniae-induced pyogenic liver abscess complain of ocular symptoms. In the majority of patients with EKE associated with pyogenic liver abscess, visual outcome is generally poor despite aggressive antibiotic therapy. Early diagnosis and prompt intervention with intravitreal antibiotics within 48 hours may salvage useful vision in some patients with EKE.

Carcinoma ex pleomorphic adenoma of the lacrimal gland: a case report.

Carcinoma ex pleomorphic adenoma affects glandular structures, occurring mostly in major salivary glands and less commonly in the lacrimal gland. We present a rare case in the lacrimal gland. We highlight the importance of keeping this rare tumor in mind and the early detection of symptoms because such a malignant transformation could occur in pleomorphic adenomas, with the proportion dependent on the duration of the tumor.