Herpetic Keratitis Research Articles

Loss of TRPM8 Exacerbate Herpes Simplex Keratitis Infection in Mice by Promoting the Infiltration of CD11b+ Ly6G+ Cells and Increasing the Viral Load in the Cornea.

To reveal the role of transient receptor potential cation subfamily M member 8 (TRPM8) channels in herpes simplex keratitis (HSK). HSK models were established using TRPM8 knockout (TRPM8-/-) mice and their wild-type (WT) littermates. The infected corneas were graded and harvested to evaluate the mRNA levels of inflammatory factors through quantitative real-time polymerase chain reaction (RT-PCR), as well as the infiltration of inflammatory cells through immunofluorescence staining and flow cytometry. Viral titers were determined by plaque assay and absolute quantitative method. RNA-sequencing was conducted to elucidate the transcriptome of corneal epithelium in response to TRPM8 knockout after infection. The anti-inflammatory effect of TRPM8 agonist menthol was documented via subconjunctival administration. Compared to their wild-type counterparts, TRPM8-deficient mice exhibited exacerbated infection symptoms and thicker corneas in HSK models. Infection in TRPM8-deficient mice resulted in significant lymphocyte infiltration, primarily consisting of Ly6G+ CD11b+ cells. Additionally, TRPM8-deficient mice displayed increased levels of corneal viral titers after infection, along with decreased expression of interferon-stimulated genes (ISGs). Subconjunctival administration of menthol effectively alleviated infection-induced symptoms and Ly6G+ CD11b+ cell infiltration in herpes simplex virus type 1 (HSV-1)-treated mice. TRPM8 promoted host resistance to HSV-1 infection by suppressing the accumulation of Ly6G+ CD11b+ cells and virus replication. These findings suggest that targeting TRPM8 could be valuable for therapeutic interventions against HSV-1 infections.

Metabolic signatures of tear extracellular vesicles caused by herpes simplex keratitis

PurposeHerpes simplex keratitis (HSK), caused by type 1 herpes simplex virus (HSV) reactivation, is a severe infectious disease that leads to vision loss. HSV can trigger metabolic reprogramming in the host cell and change the extracellular vesicles (EV) cargos; however, little is known about the EV metabolic signatures during ocular HSV infection. Here, we aimed to depict the EV-associated metabolic landscape in HSK patients’ tears. MethodsWe collected 82 samples from 41 participants with unilateral HSK (contralateral unaffected tears were set as negative control), including subtype cohorts of 13 epithelial, 20 stromal, and 8 endothelial HSK. We isolated tear EVs via our previously established platform and conducted metabolic analysis using LC-MS/MS. The metabolic signatures for recognizing HSK and subtypes were assessed through differential analysis and machine learning algorithms. ResultsHypopsia and increased extracellular CD63 levels were observed in affected eyes. We identified 339 metabolites based on sEVs isolated from tears. Differential analysis revealed alterations in energy and amino acid metabolism, as well as the infectious microenvironment. Furthermore, we observed dysregulated metabolite such as methyldopa, which is associated with inappropriate neovascularization and corneal sensation loss, contributing to the HSK severity particularly in the stromal subtype. Moreover, machine learning classification also suggested a set of EV metabolic signatures that have potential for pan-keratitis detection. ConclusionsOur findings demonstrate that tear EV metabolites can serve as valuable indicators for comprehending the underlying pathological mechanisms. This knowledge is expected to facilitate the development of liquid biopsy means and therapeutic target discovery.

Incidence of Acanthamoeba Keratitis in the United Kingdom in 2015: A Prospective National Survey.

The aims of this study were to quantify the incidence of Acanthamoeba keratitis (AK) in the United Kingdom and investigate risk factors and management parameters. This was a prospective population-based study from January to December 2015 through the British Ophthalmic Surveillance Unit. Data were collected on demographics, clinical features, and management. Incidence rates were calculated from estimates of population and contact lens (CL) user numbers. Statistical analysis compared annualized incidences per million and altered risk ratios for AK with the England and Wales 24 months 1997/1998 to 1998/1999 study. The study identified 124 AK cases, an overall incidence of 2.35 per million. CL wearers accounted for 108 of 124 cases (87%), in whom the AK incidence was 26.94 per million. Herpes keratitis was initially misdiagnosed in 25 of 124 cases (20.2%). The highest incidence of AK was among planned replacement soft CL (PRSCL) wearers (50.65 per million), 7-fold greater than for daily disposable CL (DDSCL) users (7.24 per million). There was a significant increase in AK incidence ( P < 0.001) compared with both 1997/1998 [risk ratio 1.92, 95% confidence interval (CI) 1.38-2.66] and 1998/1999 (risk ratio 2.13, 95% CI 1.52-2.98) together with a higher incidence per million CL users of 26.94 versus 21.14 (1997/1998) and 17.53 (1998/1999). This study provides the first published data on the nationwide incidence of AK in the United Kingdom. The findings confirm an increasing incidence of AK, particularly among contact lens wearers since 1997/1998 to 1998/1999. PRSCLs were identified as a significant risk factor compared with DDSCLs. Misdiagnosis and treatment delays remain an ongoing problem for patients with AK.

Опыт применения новых технологий оптической когерентной томографии в диагностике заболеваний роговицы у собак и кошек

In the article we present the results of the clinical study of cornea optical coherence tomography (OCT) in cats and dogs, performed by the authors during complex diagnostics of various keratophathies, such as: endothelial corneal dystrophy, pigmentary keratitis, chronic keratoconjunctivitis, chronic herpetic keratitis, ulcerative keratitis, corneal injuries, cats’ corneal sequestration, chronic keratitis complications of glaucoma. Normal cornea OCT scans characteristics for cats and dogs are given. OCT scans of various corneal pathologies in cats and dogs are presented and the identified pathological changes are described. Corneal OCT scan results of animals after keratoplasty operations with the use of various grafts forms have a great clinical interest. The results of the current clinical study suggest that corneal OCT in cats and dogs is highly informative and promising method of additional diagnosis of severe keratopathies, especially in cases of weak effectiveness of traditional drug therapy regimens, as well as before performing keratoplasty and in its postoperative period.

Macrodendritic ulcerative keratitis and conjunctival lymphoid hyperplasia in horses with equine herpesvirus-2 and equine herpesvirus-5 infections.

The aim of this study was to describe the clinical, confocal microscopic, histologic, and virologic features of horses with macrodendritic ulcerative keratitis and conjunctival lymphoid hyperplasia associated with equine herpesvirus-2 and equine herpesvirus-5 infection. Four foals with bilateral ocular disease. Complete ophthalmic examination was performed for each horse, and corneal samples were collected for cytology and microbiologic evaluation, including virus isolation and molecular diagnostics for the equine herpesviruses. Invivo confocal microscopy examination of the cornea was performed in two horses. Conjunctival biopsies for histopathology were collected from two horses with nodular conjunctival thickening. Each horse had bilateral, large, superficial dendritic corneal ulcerations that covered extensive regions of the corneal surface. Corneal invivo confocal microscopy examination in two horses detected inflammatory cells and populations of morphologically abnormal corneal epithelial cells adjacent to the ulcerations. The abnormal epithelial cells included round, relatively small, hyperreflective cells intermixed with elongated, enlarged, hyperreflective cells. Equine herpesvirus-2 was isolated from corneal samples of 2 horses and detected by PCR assay in the other two horses. Equine herpesvirus-5 was also detected by PCR assay in three of the horses. Conjunctival histopathology identified predominantly lymphocytic infiltrates. The macrodendrites and conjunctival masses resolved with topical antiviral therapy (cidofovir or idoxuridine) in all horses and did not recur. The equine gammaherpesviruses may be associated with the development of macrodendritic ulcerative keratitis and conjunctival lymphocytic masses in foals. Invivo confocal microscopy of horses with macrodendrites revealed similar findings to other host species with herpetic dendritic keratitis.

Diagnostic performance of real-time quantitative PCR in tear samples in various subtypes of herpes simplex keratitis.

Diagnosis of herpes simplex keratitis (HSK) is mostly based on clinical findings, yet biological confirmation supports management of challenging cases. This study evaluated the place of real-time quantitative PCR (RT-qPCR) on tear samplings in the management of HSK. Clinical records of patients who underwent tear sampling tested by RT-qPCR for herpes simplex virus type 1 for an acute episode of corneal inflammation or defect between January 2013 and December 2021 were retrospectively reviewed, and results were compared to clinical diagnosis (i.e., HSK or not) based on biomicroscopic findings and medical history. Of 465 tested tear samples from 364 patients, a clinical diagnosis of active (ongoing) HSK was recorded in 240 cases, among which 76 were RT-qPCR positive (global sensitivity of 31.6%, specificity of 99.5%). Sensitivity of RT-qPCR was higher in epithelial (97.4%) and stromal keratitis with ulceration (48.7%), compared to other types of HSK (23.5% in keratouveitis, 13.6% in endotheliitis, 11.1% in postherpetic neurotrophic keratopathy, and 8.1% in stromal keratitis without ulceration). The highest viral loads were detected from epithelial and stromal keratitis with ulceration, while in HSK with no epithelial involvement, the viral load detected was 196-fold lower, on average. The proportion of clinically characterized HSK patients with negative tear samples was higher in patients receiving antiviral treatment (P < 0.0001). RT-qPCR, performed on tear samples, can help in confirming diagnosis in case of presumed HSK, including clinical forms with no obvious epithelial involvement. The sensitivity of tear sampling is much higher whenever epithelial keratitis is present.

Targeted delivery of herpes simplex virus glycoprotein D to CD169+ macrophages using ganglioside liposomes alleviates herpes simplex keratitis in mice

Herpes simplex keratitis (HSK) is a common blinding corneal disease caused by herpes simplex virus type 1 (HSV-1) infection. Antiviral drugs and corticosteroids haven't shown adequate therapeutic efficacy. During the early stage of HSV-1 infection, macrophages serve as the first line of defense. In particular, CD169+ macrophages play an important role in phagocytosis and antigen presentation. Therefore, we constructed GM-gD-lip, a ganglioside GM1 liposome vaccine encapsulating HSV-1 glycoprotein D and targeting CD169+ macrophages. After subconjunctival injection of the vaccine, we evaluated the survival rate and ocular surface lesions of the HSK mice, as well as the virus levels in the tear fluid, corneas, and trigeminal ganglia. We discovered that GM-gD-lip reduced HSV-1 viral load and alleviated the clinical severity of HSK. The GM-gD-lip also increased the number of corneal infiltrating macrophages, especially CD169+ macrophages, and polarized them toward M1. Furthermore, the number of dendritic cells (DCs) and CD8+ T cells in the ocular draining lymph nodes was significantly increased. These findings demonstrated that GM-gD-lip polarized CD169+ macrophages toward M1 to eliminate the virus while cross-presenting antigens to CD8+ T cells via DCs to activate adaptive immunity, ultimately attenuating the severity of HSK. The use of GM-gD-lip as an immunotherapeutic method for the treatment of HSK has significant implications.

Design and Optimization of Ganciclovir Nanosuspension Loaded In Situ Gelling Mucoadhesive Eye Drops for Herpetic Keratitis

Ganciclovir (GCR), an antiviral drug used to treat herpetic keratitis, is less effective because of its poor bioavailability (BCS III). Nanosuspension (NS) is a promising technique for improving solubility and the dissolution of poorly soluble drugs by size reduction. Topical solution or conventional eye drop represents the easiest route to deliver drugs to the anterior eye segment; however, low precorneal retention, high nasolacrimal drainage, and metabolic degradation lead to low bioavailability. To overcome these limitations, incorporating GCR-NS into mucoadhesive in situ gel-forming (ISG) could enhance its ocular permeability and drug bioavailability. Therefore, this study aimed to design and optimize GCR-NS-loaded in situ gelling mucoadhesive eye drops (GCR-NS-loaded mucoadhesive ISG) for improving ophthalmic delivery. Pluronic® F127 (P127), an ophthalmic gel forming, was selected as a polymer due to self-gelling formation by temperature-triggered in situ gelling at ocular temperature. Moreover, hyaluronic acid-modified catechol (HA-cat), a novel mucoadhesive polymer, was combined to achieve the desired ocular mucoadhesion and facilitate ophthalmic delivery. Optimized GCR-NS formulation prepared using the nanoprecipitation technique was selected based on the central composite design (CCD) model by evaluating their particle size, polydispersity index (PDI), and zeta potential, before being loaded into mucoadhesive ISG. The optimal GCR-NS-loaded mucoadhesive ISG (F7) revealed the desired physicochemical properties with better viscosity, mucoadhesion, and gelling capacity at physiological conditions, high ocular permeation with a sustained manner over 24 h compared to eye drop suspension. Accordingly, GCR-NS-loaded mucoadhesive ISG could be a promising ocular delivery system for the effective local delivery of GCR for herpetic keratitis. Keywords: Ganciclovir, Nanosuspension, Mucoadhesive eye drop, Ophthalmic delivery, In situ gelling

Эффективность физических методов лечения рецидивирующей эрозии роговицы герпетической этиологии

94 patients were treated with recurrent corneal erosion of herpes genesis on the background of antiviral, reparative therapy. Physical therapies were added to the baseline therapy to stimulate reparative processes. The following methods of treatment were used: exposure to local reduced pressure, magnetic therapy, vitamin-protein baths. In a comparative analysis with a control group, results were obtained that allow reducing the treatment time, restoring corneal transparency «to achieve a high optical effect. All patients had good tolerability, no adverse reactions, and no surgical interventions were required. Keywords: herpetic keratitis, physical methods of treatment

The effectiveness of a new antiviral drug of plant origin in the form of eye drops in the complex therapy of ophthalmic herpes

Ophthalmic herpes is the most common infection-related cause of blindness in developing countries.Purpose: to evaluate the therapeutic efficacy, safety and tolerability of the drug Panavir® used as part of a combination therapy for herpetic superficial dendritic keratitis.Materials and methods. The study involved 106 patients aged 18 to 65 years divided into two groups. Group 1 (54 patients; 54 eyes) received treatment with Panavir® and Acyclovir ophthalmic ointment, whilst Group 2 (52 patients; 52 eyes) received a placebo of Panavir® and Acyclovir ophthalmic ointment. The primary parameter of the efficiency was the percentage of patients achieving clinical recovery, i.e. signs of complete epithelialization of the cornea.Results. In group 2 (monotherapy), a significantly greater number of patients with mild conjunctival hyperemia was registered than in the combination therapy group 1 — 59.26 vs. 23.08 %, p &lt; 0.001. In group 1, the proportion of patients without corneal syndrome at visits 3 and 4 was, respectively, 11.11 and 61.11 %, while in group 2 it was only 3.85 and 19.61 %, respectively.Conclusion. The drug Panavir® eye drops 0.004 % showed a high therapeutic efficiency in the combined treatment of herpetic keratitis, and good tolerance. It also reduced the time required for corneal epithelialization and conjunctiva normalization.

The Orphan Drug for Acanthamoeba Keratitis (ODAK) Trial: PHMB 0.08% (Polihexanide) and Placebo versus PHMB 0.02% and Propamidine 0.1%

ObjectivesTo compare topical PHMB (polihexanide) 0.02% (0.2 mg/ml) + propamidine 0.1% (1 mg/ml) [PHMB 0.02%+propamidine] to PHMB 0.08% (0.8 mg/ml) with placebo [PHMB 0.08%] for Acanthamoeba keratitis (AK) treatment. DesignProspective randomized, double-masked, active-controlled, multicentre, Phase3 study (ClinicalTrials.gov NCT03274895). Participants135 at six European centres between 08/17/2017 to 06/18/2021. MethodsPrincipal inclusion criteria: ≥12 years old; in vivo confocal microscopy (IVCM) with clinical findings consistent with AK. Also included were participants with concurrent bacterial keratitis, using topical steroids, antiviral and antifungal drugs before randomisation. Principal exclusions: concurrent herpes or fungal keratitis, use of anti-amoebic therapy (AAT). Randomisation: 1:1 computer-generated, block size 4. This was a superiority trial having a predefined non-inferiority margin. The sample size of 130 participants gave approximately 80% power to detect 20 percentage point superiority for PHMB 0.08% for the primary outcome of the medical cure rate (without surgery or change of AAT) within 12 months (MCR_12), cure defined by clinical criteria 90 days after discontinuing anti-inflammatories and AAT. A prespecified multivariable analysis adjusted for baseline imbalances in risk factors affecting outcomes. Main outcome measuresThe MCR_12. Secondary outcomes included best-corrected visual acuity (BCVA) and treatment failure rates. Safety outcomes included adverse event rates. Results135 participants were randomised providing 127 in the full analysis subset (61 on PHMB 0.02%+propamidine and 66 on PHMB 0.08%) and 134 in the safety analysis subset. The adjusted MCR_12 was 86.6% (unadjusted 88.5%) for PHMB 0.02%+propamidine and 86.7% (unadjusted 84.9%) for PHMB 0.08%; the non-inferiority requirement for PHMB 0.08% was met (adjusted difference 0.1 percentage points, lower one-sided 95% confidence limit -8.3 percentage points). Secondary outcomes were similar for both treatments and not analysed statistically: median BCVA of 20/20, an overall treatment failure rate of 17/127 (13.4%) of whom 8/127 (6.3%) required therapeutic keratoplasty. There were no serious drug related adverse events. ConclusionsPHMB 0.08% monotherapy may be as effective (or at worse only eight percentage points less effective) as dual therapy with PHMB 0.02%+propamidine (a widely used therapy) with medical cure rates of >86%, when used with the Trial treatment delivery protocol, in AK populations with similar disease severity.

Headache associated with travoprost

Background: Putative adverse effects of prostaglandin analogs in patients treated for glaucoma include periocular skin darkening, herpes simplex keratitis, cystoid macular edema, iritis, and headaches. Here, we report a case of migraine headache after a travoprost administration and discuss the role of prostaglandin analogs in migraine-like headaches.&#x0D; Case Presentation: A 70-year-old man visited the hospital complaining of pain, redness, and sensitivity to light for 1 week associated with gradual loss of vision in the right eye after cataract surgery, which had been performed 2 years prior. After the examination, the patient was diagnosed with pseudophakic bullous keratopathy and advised to undergo optical penetrating keratoplasty of the right eye. On postoperative day 3, his intraocular pressure (IOP) was 30 mmHg at 10:00 AM in the right eye. A single dose of 0.004% travoprost was instilled in the right eye on the same day. His IOP decreased to 16 mmHg at 2:00 PM. The next day, he presented with migraine-like headache that had started within 1 h after the instillation the previous night. Unremarkable neurological examination and neuroimaging suggested that travoprost had caused the migraine. Although the headache gradually resolved, it reappeared after the administration of other prostaglandins. Trabeculectomy was performed on the right side. The IOP was controlled, and the headache was resolved.&#x0D; Conclusions: This case indicates a potential causal relationship between topical prostaglandin analogs and migraine-like headaches, as evidenced by symptom resolution upon discontinuation. However, large-scale studies including control groups are required to prove a causal relationship between topical prostaglandin analog administrations and headache development.

Herpes simplex keratitis following coronavirus disease 2019 vaccination: an observational study of forty-three cases with different presentations

Objective The aim of this study was to evaluate the cases diagnosed with different presentations of herpes simplex virus (HSV) keratitis following coronavirus disease 2019 (COVID-19) vaccination and to describe the relationship between the disease presentation, the type of vaccine, and the duration between vaccination and disease onset. Patients and methods This is a retrospective observational clinical study of HSV keratitis cases that was carried out by the Cornea Team at Alexandria Ophthalmology Hospital, Alexandria, Egypt, between March and September 2022. Full medical and ophthalmic history was taken, followed by a thorough ophthalmic examination. The vaccination data of the patients were recorded. The hospital records of the patients visiting the outpatient clinics, at that period, were collected regarding the number of vaccinated and nonvaccinated patients and the type of vaccine received. Results The study included 43 eyes of 43 patients. Typical dendritic corneal ulcer occurred in 21 (48.8%) eyes, while 5 (11.6%) eyes presented with a geographic ulcer, and 14 (32.6%) eyes were diagnosed as interstitial keratitis with or without ulceration. The remaining 3 (6.98%) eyes presented with disciform keratitis. Vaccination was previously received by 40 (93.02%) patients, while only 3 patients did not receive any vaccine. Oxford-Astra Zeneca vaccine was received by 14 (35%) patients, while the Pfizer-BioNTech vaccine was received by 13 (32.5%) patients, Sinovac vaccine by 10 (25%) patients, and other types of vaccines by three (7.5%) patients. The duration between the last dose received from the vaccine and the onset of symptoms ranged between 2 and 58 days (mean=18.35±13.3 days). Reviewing the hospital records, out of the 65 321 patients who visited the hospital during that period, 47% were nonvaccinated, and 53% were vaccinated with one dose or more of COVID-19 vaccine (Odds ratio=11.8207 indicating a significant association between HSV keratitis and vaccination). Conclusion Reactivation of HSV keratitis may follow COVID-19 vaccination with different presentations of herpetic keratitis. The Oxford-AstraZeneca and the Pfizer-BioNTech vaccines were significantly associated with HSV ocular infection more than any other type of vaccine.

Ocular Gene Therapy: Literature Review

This literature offers complete information about the advances in gene treatment in the of the eye, inclusive of cornea, conjunctiva, lacrimal gland, and trabecular meshwork. We talked about gene transport systems, collectively with viral and non-viral vectors as exact as gene modifying techniques, generally CRISPR-Cas9, and epigenetic treatments, consisting of antisense and siRNA therapeutics. We moreover furnish a specific assessment of gene treatment has been examined with corresponding outcomes. Disease stipulations embody corneal and conjunctival fibrosis and scarring, corneal epithelial wound healing, corneal graft survival, corneal neovascularization, genetic corneal dystrophies, herpetic keratitis, glaucoma, dry eye disease, and specific ocular surface diseases. Although most of the analyzed consequences on the use and validity of gene treatment at the ocular surface have been offered in vitro or the utilization of animal models, we moreover talked about the on hand human studies. Gene treatment methods are nowadays viewed very promising as rising future remedies of a variety of diseases, and this area is unexpectedly expanding.

Herpes simplex virus keratitis: electronic medical records driven big data analytics report from a tertiary eye institute of South India.

To describe the demographics and clinical profile of Herpes Simplex Virus (HSV) Keratitis in patients presenting to a multi-tier ophthalmology hospital network in South India. We have reviewed the medical records of all patients having a clinical diagnosis of any form of HSV keratitis, seen between May 2012 and August 2020 across the L V Prasad Eye Institute network. All the further analyses of the groups were performed using the keywords used for making the diagnosis of HSV keratitis and the data were collected from the electronic medical record system. There were a total of 8308 (N = 8897 eyes) patients. Male: female ratio was 5368 (64.61%):2940 (35.39%). Unilateral involvement was in 7719 (92.91%) patients. The most common age group affected was between the third to fifth decades of life with 1544 (18.58%). 3708 (1.68%) eyes had mild visual impairment (< 20/70) while the rest of them had moderate to severe visual impairment as observed mainly (p ≤ 0.01) in Necrotizing stromal keratitis. 7314 (82.21%) eyes had normal intraocular pressure (10-21mm Hg) while raised most commonly in keratouveitis (P ≤ 0.01). Epithelial Keratitis, Immune Stromal Keratitis, Endotheliitis, Neurotrophic keratopathy and Keratouveitis were observed in 1875 (17.22%) eyes, 5430 (61.03%) eyes, in 129(1.45%) eyes, 1188 (13.35%) eyes, 148 (1.66%) eyes and 256 (2.88%) eyes respectively. Based on our institute-based data, the most common type of HSV keratitis is Immune stromal keratitis followed by epithelial keratitis. Although not representative of the general population, this data provide useful insights related to HSV keratitis from India.

Therapeutic Potential of Biochanin A in Herpes Simplex Keratitis.

Herpes simplex keratitis (HSK) is a blinding eye disease that is initiated by the herpes simplex virus type 1 (HSV-1). Resistance to acyclovir (ACV) and the side effects of corticosteroid drugs have become concerning issues, so it is crucial to develop new antivirals for treating HSK. In this study, we report that biochanin A (BCA), a naturally occurring flavonoid compound, provides multifaceted protective effects with anti-viral, anti-inflammatory, anti-oxidative stress and anti-apoptotic activities to alleviate HSK. The results show that BCA significantly inhibited HSV-1 replication in vitro and further proved that BCA principally influenced the early stage of virus infection. We reveal that BCA downregulated the expression of pro-inflammatory factors triggered by HSV-1, including TNF-α, RANTES, IL-1β and IL-6. Furthermore, BCA treatment alleviated oxidative stress and apoptotic arising from HSV-1 infection. Lastly, we induced HSK in male C57BL/6 mice and treated them with either BCA or phosphate buffer solution (PBS) eye drops. We observed the ocular surface lesions; determined the virus load in the tear fluid, corneas as well as trigeminal ganglions (TGs); and detected the levels of inflammation and apoptosis in the corneas simultaneously. These results show that BCA inhibits HSV-1 and alleviates the corneal lesion degree. Our study illustrates that BCA is a promising therapeutic approach for application in treating HSK.

Clinical Spectrum of Herpes Simplex Keratitis in a Tertiary Eye Hospital, Nepal

Background: Ocular manifestations of herpes simplex virus infection range from benign blepharitis, conjunctivitis, and epithelial keratitis to vision-threatening stromal keratitis and uveitis. This study aims to find out the demographic profile, clinical manifestations of herpes simplex virus keratitis and to assess associated visual impairment in Nepalese population. Method: The retrospective study was conducted from January 2020 – February 2022. The medical records were reviewed where visual acuity at presentation and at one month follow-up, clinical details on examination and, diagnosis of the stage of disease were noted. Results: Of 112 herpes simplex keratitis patients, mean age was 47.19±19.14 years. Patients of herpes simplex keratiris presented with different types of clinical manifestations namely 25 (22.31%) cases were dendritic and 13 (11.6%) geographic ulcers, 38 (33.9%) cases presented with purely stromal lesions, 19 (16.9%) cases with epithelial plus stromal lesions, 11 (9.8%) cases with stromal plus endothelial lesions and six (5.3%) cases with endothelitis. At presentation, 33 (29.4) had visual acuity of 6/6–6/18, 50 (44.6%) had 6/18–6/60, 6 (5.3%) had 6/60–3/60 and 23 (20.5%) had &lt;3/60. At one month follow up 71 (63.3%) had visual acuity of 6/6–6/18, 30 (26.7%) had 6/18–6/60, 5 (4.4%) had &lt;6/60–3/60 and 6 (5.35%) had &lt;3/60. Conclusion: Herpes simplex keratitis has diverse presentation and can cause significant vision impairment. Early clinical diagnosis and management may prevent disease progression and visual impairment.

In vivo CRISPR gene editing in patients with herpetic stromal keratitis

Invivo CRISPR gene therapy holds large clinical potential, but the safety and efficacy remain largely unknown. Here, we injected a single dose of herpes simplex virus 1 (HSV-1)-targeting CRISPR formulation in the cornea of three patients with severe refractory herpetic stromal keratitis (HSK) during corneal transplantation. Our study is an investigator-initiated, open-label, single-arm, non-randomized interventional trial at a single center (NCT04560790). We found neither detectable CRISPR-induced off-target cleavages by GUIDE-seq nor systemic adverse events for 18months on average in all three patients. The HSV-1 remained undetectable during the study. Our preliminary clinical results suggest that invivo gene editing targeting the HSV-1 genome holds acceptable safety as a potential therapy for HSK.

Mechanistic of Vesicular Ethosomes and Elastic Liposomes on Permeation Profiles of Acyclovir across Artificial Membrane, Human Cultured EpiDerm, and Rat Skin: In Vitro-Ex Vivo Study.

Acyclovir (ACV) controls cutaneous herpes, genital herpes, herpes keratitis, varicella zoster, and chickenpox. From previously reported ACV formulations, we continued to explore the permeation behavior of the optimized ACV loaded optimized ethosome (ETHO2R) and elastic liposome (ELP3R) and their respective carbopol gels across artificial membrane, cultured human EpiDerm, and rat skin. Transepidermal water loss (TEWL), scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), and atomic force microscopy (AFM) were used to investigate the mechanistic perspective of permeation behavior. The size values of reformulated ELP3-R and ETHO2-R were observed as 217 and 128 nm, respectively (close to previous report), whereas their respective gels showed as 231 and 252 nm, respectively. ETHO2R showed high elasticity, %EE, and low vesicle size. These were investigated for the diffusion rate of the drug permeation (3 h) across the artificial membrane, cultured human EpiDerm, and rat skin. ETHO2GR showed the highest permeation flux (78.42 µg/cm2/h), diffusion coefficient (8.24 × 10-5 cm2/h), and permeation coefficient (0.67 × 10-3 cm/h) of ACV across synthetic membrane, whereas diffusion coefficient (2.4 × 10-4 cm2/h) and permeation coefficient (0.8 × 10-3 cm/h) were maximum across EpiDerm for ETHO2GR. ETHO2R suspension showed maximized permeation flux (169.58 µg/cm2/h) and diffusion rate (0.293 mg/cm2/h1/2), suggesting the rapid internalization of vesicles with cultured skin cells at low viscosity. A similar observation was revealed using rat skin, wherein the permeation flux (182.42 µg/cm2/h), permeation coefficient (0.3 × 10-2 cm/h), and diffusion rate (0.315 mg/cm2/h1/2) of ETHO2R were relatively higher than ELP3R and ELP3GR. Relative small size (128 nm), low viscosity, ethanol-mediated ultra-deformability, high drug entrapment (98%), and elasticity (63.2) are associated with ETHO2R to provide remarkable permeation behavior across the three barriers. The value of TEWL for ETHO2R (21.9 g/m2h) was 3.71 times higher than untreated control (5.9 g/m2h), indicating ethanol-mediated maximized surficial skin lipid perturbation at 3 h of application, whereas the respective ETHO2GR-treated rat skin had TEWL value (18.6 g/m2h) slightly lower than ETHO2R due to gel-based hydration into the skin. SEL, CLSM, and AFM provided a mechanistic perspective of ETHO2R and ELP3R-mediated permeation across rat skin and carrier-mediated visualization (skin-vesicle interaction). AFM provided detailed nanoscale surface roughness topographical parameters of treated and untreated rat skin as supportive data to SEM and CLSM. Thus, ethosomes ETHO2R and respective gel assisted maximum permeation of ACV across rat skin and cultured human EpiDerm to control cutaneous herpes infection and herpes keratitis.

Luteolin inhibits herpes simplex virus 1 infection by activating cyclic guanosine monophosphate-adenosine monophosphate synthase-mediated antiviral innate immunity

BackgroundThe successive outbreaks of large-scale infectious diseases due to virus infection have been a major threat to human health in recent decades. Herpes simplex virus I (HSV-1) is a widely-disseminated DNA virus that infects the central nervous system to cause herpes labialis, keratitis and herpes simplex virus encephalitis (HSE), resulting in recurrent lifelong clinical or subclinical episodes. Luteolin is a plant flavone that has been extensively used in the treatment of various human diseases, including carcinogenesis, inflammation and chronic degenerative diseases. PurposeThe aim of this study was to investigate the antiviral molecular mechanism of luteolin against HSV-1 infection in vitro and in vivo. MethodsThe antiviral effect of luteolin in cell lines was examined by viral plaque assay, RT-qPCR, Western blot and time-of-addition assay. The interaction between luteolin and cyclic guanosine monophosphate-adenosine monophosphate synthase (cGAS) was evaluated by molecular modeling and semi-denaturing detergent agarose gel electrophoresis. The efficacy of luteolin on HSE was evaluated in the HSE mouse model by analyzing weight loss, neurodegenerative symptoms and histopathological scores. Cytokine expression and protein levels were examined by RT-qPCR, Western blot and ELISA. ResultsLuteolin inhibited the early process of HSV-1 infection, without affecting the infection of acyclovir-resistant HSV-1 strains. In addition, luteolin enhanced antiviral type I interferon production and activated the cytoplasmic DNA-sensing cGAS-stimulator of interferon gene (STING) pathway. Luteolin directly bound the active substrate binding site and promoted the oligomerization of cGAS. Luteolin also inhibited HSE-related weight loss, neurodegeneration and neuroinflammation in mice caused by HSV-1 infection. Furthermore, luteolin enhanced type I interferon expression and stimulated the cGAS-STING signaling pathway in vivo. ConclusionLuteolin inhibited the post-entry process of HSV-1 by activating the cGAS-STING pathway to promote antiviral interferon production. These results provided the rationale for luteolin as a potent cGAS activator and antiviral agent.