Acanthamoeba keratitis: perspectives for patients

Purpose: To unveil the long-term prognosis of Acanthamoeba keratitis based on clinical presentation and timing of diagnosis to better inform patients since the first visit regarding their length of treatment, quality of life, and visual function.Methods: Retrospective observational study enrolling patients with Acanthamoeba keratitis from 1994 to 2019. Patients with a complete eye examination and medical records were analyzed. The severity of the disease, the time from onset of symptoms to the appropriate therapeutic regimen, the time until clinical resolution, visual function, and long term follow-up was evaluated. Quality of life was assessed at the last follow-up visit by means of the VFQ-25 questionnaire.Results: Thirty-five patients (40 eyes) were assessed. The overall healing time of patients with Acanthamoeba keratitis was 12.5 ± 3.5 months, while patients with a severe corneal ulcer (stage III) had a significant longer healing time (16.2 ± 3.7 months) compared to patients with stage II (7.04 ± 0.7 months) or I (7.7 ± 1.5 months; p < .05). Patients who received a prompt therapy (<30 days form symptoms onset) had a reduced healing time compared to patients with a delayed diagnosis (p < .01). Quality of life was assessed after a mean of 11.7 ± 4.7 years and it was mildly reduced (86.6 ± 17). Patients that were diagnosed early (<30 days from onset) showed a lower reduction in quality of life than in patients that were diagnosed >30 days from onset. After resolution, 59% of the patients considered unnecessary any further proposed surgical intervention.Conclusions: Delayed diagnosis of Acanthamoeba keratitis and disease severity significantly increases healing time and duration of treatment. The time to diagnosis and disease stage at diagnosis predicts the duration of treatment, the final outcome, quality of life, and the requirement of surgery. These data would allow us to promptly inform patients about long-term disease timeline, future outcomes, improving disease acceptance, and quality of life

Sands of Sahara after LASIK in Avellino Corneal Dystrophy

We report the case of a patient diagnosed with Avellino corneal dystrophy (ACD) who developed diffuse interstitial keratitis following excimer laser insitu keratomileusis (LASIK). ACD is an autosomal dominant corneal dystrophy characterized by multiple asymmetric stromal opacities that impair vision. Accepted treatments for this condition include corneal transplantation and phototherapeutic keratectomy (PTK). Our patient underwent LASIK at another institution to correct myopia. LASIK and photorefractive keratectomy (PRK) are usually contraindicated in ACD for the high risk of disease recurrence and postoperative complications. The patient came to our attention lamenting blurry vision, decreased visual acuity, and photophobia. Ophthalmologic examination revealed bilateral interstitial keratitis, also known as “sands of Sahara”, a seldom-seen complication of LASIK characterized by fine and diffuse granular infiltrates at the surgical flap interface.The risk of developing interstitial keratitis, as in the case presented here, represents another valid reason for avoiding LASIK in patients with ACD

NGF modulates trkANGFR/p75NTR in αsMA-expressing conjunctival fibroblasts from human ocular cicatricial pemphigoid (OCP)

OBJECTIVE: In a previous study, we reported the upregulation of Nerve Growth Factor (NGF) and trkANGFR expression in Ocular Cicatricial Pemphigoid (OCP), an inflammatory and remodeling eye disease. Herein, we hypothesize a potential NGF-driven mechanism on fibroblasts (FBs) during OCP remodeling events. To verify, human derived OCP-FBs were isolated and characterized either at baseline or after NGF exposure. MATERIALS AND METHODS: Conjunctival biopsies were obtained from 7 patients having OCP and 6 control subjects (cataract surgery). Both conjunctivas and primary FB cultures were characterised for αSMA, NGF and trkANGFR/p75NTR expression. Subcultures were exposed to NGF and evaluated for αSMA, NGF, trkANGFR/p75NTR expression as well as TGFβ1/IL4 release. For analysis, early and advanced subgroups were defined according to clinical parameters. RESULTS: OCP-conjunctivas showed αSMA-expressing FBs and high NGF levels. Advanced OCP-FBs showed higher αSMA expression associated with higher p75NTR and lower trkANGFR expression, as compared to early counterparts. αSMA expression was in keeping with disease severity and correlated to p75NTR. NGF exposure did not affect trkANGFR levels in early OCP-FBs while decreased both αSMA/p75NTR expression and TGFβ1/IL4 release. These effects were not observed in advanced OCP-FBs. CONCLUSIONS: Taken together, these data are suggestive for a NGF/p75NTR task in the potential modulation of OCP fibrosis and encourages further studies to fully understand the underlying mechanism occurring in fibrosis. NGF/p75NTR might be viewed as a potential therapeutic target

Lamellar macular holes: monitoring and management strategies

Lamellar macular holes are a vitreoretinal condition characterized by abnormalities in foveal contour with splitting of the neuroepithelium and often an intact photoreceptor layer. Recent developments in high-resolution imaging have increased our ability to study the details of the vitreoretinal interface and to distinguish between different forms of lamellar holes. A new classification is needed to help clinicians in the management of lamellar macular holes. Some clinicians prefer to observe these clinical entities, especially when visual acuity is maintained or alterations of the photoreceptor layer are present. Nevertheless, lamellar holes may sometimes progress, and visual acuity can deteriorate. On the other hand, surgical treatment may lead to positive anatomical and functional outcomes, but not without risks. This review provides a critical overview of the available data on lamellar macular holes, focusing on diagnosis and managing options

Ocular surgical models for immune and angiogenic responses

Corneal transplantation serves as a reproducible and simple surgical model to study mechanisms regulating immunity and angiogenesis. The simplicity of the model allows for systematic analysis of different mechanisms involved in immune and angiogenic privilege and their failures. This protocol describes how to induce neovessels and inflammation in an actively regulated avascular and immune-privileged site. This involves placing intra-stromal corneal sutures for two weeks, disrupting the privileges, and performing corneal transplantation subsequently. Privileged and non-privileged recipient responses to donor cornea can be compared to identify key immunological mechanisms that underlie angiogenesis and graft rejection. This protocol can also be adapted to the growing repertoire of genetic models available in the mouse, and is a valuable tool to elucidate molecular mechanisms mediating acceptance or failure of corneal graft. The model could be used to assess the potential of therapeutic molecules to enhance graft survival in vivo

Impaired Function of Peripherally Induced Regulatory T Cells in Hosts at High Risk of Graft Rejection

Regulatory T cells (Tregs) are crucial for allograft survival. Tregs can be divided into thymus-derived natural Tregs (tTregs) and peripherally-derived induced Tregs (pTregs). Here, we determine whether the suppressive function of Treg subsets is hampered in hosts who are at high risk for rejecting their graft. To induce graft beds that promote high risk of transplant rejection, intrastromal corneal sutures were placed two weeks prior to the transplant procedure in mice. We demonstrate that in high-risk recipients the frequencies and function of pTregs (but not tTregs) are suppressed. Reduced function of pTregs correlated with decreased expression of CTLA-4, interleukin-10, and transforming growth factor-β. Adoptive transfer of pTregs from mice at low risk of subsequent graft rejection is able to rescue graft survival in recipients that are at high risk of rejecting their grafts. Our data suggest that impaired function of pTregs, but not tTregs, mediates the loss of immune tolerance and promotes allograft rejection

Challenges in acanthamoeba keratitis: a review

To review challenges in the diagnosis and management of Acanthamoeba keratitis (AK), along with prognostic factors, in order to help ophthalmologists avoid misdiagnosis, protracted treatment periods, and long-term negative sequelae, with an overarching goal of improving patient outcomes and quality of life, we examined AK studies published between January 1998 and December 2019. All manuscripts describing clinical manifestations, diagnosis, treatment, prognosis, and challenges in short- and long-term management were included. The diagnosis of AK is often challenging. An increased time between symptom onset and the initiation of appropriate therapy is associated with poorer visual outcomes. The timely initiation of standardized antiamoebic therapies improves visual outcomes, decreases the duration of treatment, and reduces the chances of needing surgical intervention. In clinical practice, AK diagnosis is often missed or delayed, leading to poorer final visual outcomes and a negative impact on patient morbidity and quality of life

Inflammaging at ocular surface: clinical and biomolecular analyses in healthy volunteers

PURPOSE. To assess the ocular surface in volunteers who consider themselves as healthy, in order to evaluate how para-inflammatory mechanisms fail with age, and thus investigate the phenomenon of "InflammAging.''METHODS. In this observational prospective cohort study, volunteers were categorized into three groups according to age: young (19-40 years), middle-aged (41-60 years), and older adults (61-93 years). Clinical assessments included tear breakup time (T-BUT) and Schirmer test type I. Dry eye symptoms were evaluated by the Ocular Surface Disease Index (OSDI) questionnaire. Conjunctival mRNA and protein expression of intercellular adhesion molecule-1 (ICAM-1), MUC5AC, and IL-8 were measured by real-time PCR and immunofluorescence.RESULTS. A total of 82 volunteers (38 males and 44 females) were enrolled. T-BUT decreased significantly with increasing age (young: 11.13 +/- 0.18 seconds; middle-aged: 10.83 +/- 0.56 seconds; older: 9.00 +/- 1.00 seconds, P < 0.05). Schirmer test values decreased significantly with age (young: 20.6 +/- 1.0 mm; middle-aged: 19.2 +/- 1.2 mm; older: 16.0 +/- 1.1 mm, P < 0.05). OSDI scores increased with age in both groups, but they were substantially higher in women. Conjunctival expression of inflammatory markers ICAM-1, IL-8, and MUC5AC increased with age.CONCLUSIONS. Clinical signs, symptoms, and biomarkers of chronic inflammation increased with age in a cohort of volunteers who considered themselves healthy, indicating an age-related progressive impairment of ocular surface system function

Age-Related Changes to Human Tear Composition.

Purpose We characterize age-associated alterations in the expression of inflammatory mediators and tissue remodeling factors in human tears. Methods A total of 75 consecutive volunteers (32 male/44 female; 19-93 years) underwent clinical assessment of ocular surface status, ocular surface disease index (OSDI) grading and tear sampling. The volunteers were categorized into three groups: young (18-40 years), middle-aged (41-60 years), and old (>60 years). Total protein profiles and chip-based protein array evaluations were conducted to investigate the expression of 60 potential candidates, including pro-/anti-inflammatory mediators and tissue remodeling factors. Appropriate validations were performed using conventional assays. Multiple comparisons for regression between potential candidates and age were performed, as well as statistical analyses among the three age groups. Nonpooled samples were used for quantifications. Results Pearson analysis of chip-arrays identified 9 of 60 potential candidates. Specifically, IL-8, IL-6, and regulated on activation, normal T cell expressed and secreted (RANTES; P < 0.0083) protein as well as matrix metalloproteinase (MMP)-1, IL-3, and TNF-α (P < 0.05) correlated positively with aging. MIP-3β showed an opposite tendency. Western blot and ELISA analysis corroborated the array data. OSDI grading did not correlate with aging. Conclusions Dynamic changes to tear protein profiles occur with aging. Our study identifies the expression of IL-8, IL-6, RANTES, MMP-1, and MIP-3β as increasing with age. These select inflammatory and matrix remodeling factors may be relevant to the development of novel diagnostic tools and therapeutics in the context of age-related ocular surface disease

T-helper 17 lymphocytes in ocular cicatricial pemphigoid

T-helper 17 lymphocytes (Th17) were identified in the healthy conjunctiva and in patients with ocular cicatricial pemphigoid (OCP), a disease characterized by chronic ocular surface inflammation