Corneal ring infiltrate- far more than Acanthamoeba keratitis: review of pathophysiology, morphology, differential diagnosis and management.

OBJECTIVE: Ring infiltrates usually accompany numerous infectious and sterile ocular disorders. Nevertheless, systemic conditions, drugs toxicity and contact lens wear may present with corneal ring infiltrate in substantial part. Considering its detrimental effect on vision, detailed knowledge on etiology, pathophysiology, differential diagnosis, and management should be considered essential for every ophthalmologist. METHODS: The PUBMED database was searched for "corneal ring infiltrate" and "ring infiltrate" phrases, "sterile corneal infiltrate" and "corneal infiltrate". We analyzed articles written in English on risk factors, pathophysiology, clinical manifestation, morphological features, ancillary tests (anterior-segment optical coherence tomography, corneal scraping, in vivo confocal microscopy), differential diagnosis and management of corneal ring infiltrate. RESULTS: Available literature depicts multifactorial origin of corneal ring infiltrate. Dual immunological pathophysiology, involving both antibodies-dependent and -independent complement activation, is underlined. Furthermore, we found that the worldwide most prevalent among non-infectious and infectious ring infiltrates are ring infiltrates related to contact-lens wear and bacterial keratitis respectively. Despite low incidence of Acanthamoeba keratitis, it manifests with corneal ring infiltrate with the highest proportion of the affected patients (one third). However, similar ring infiltrate might appear as a first sign of general diseases manifestation and require targeted treatment. Every corneal ring infiltrate with compromised epithelium should be scraped and treat as an infectious infiltrate until not proven otherwise. Of note, microbiological ulcer might also lead to immunological ring and therefore require anti-inflammatory treatment. CONCLUSION: Corneal ring infiltrate might be triggered not only by ocular infectious and non-infectious factors, but also by systemic conditions. Clinical assessment is crucial for empirical diagnosis. Furthermore, treatment is targeted towards the underlying condition but should begin with anti-infectious regimen until not proven otherwise.

Glaucoma patients have an increased level of trimethylamine, a toxic product of gut bacteria, in the aqueous humor: a pilot study.

PURPOSE: Animal studies suggest that gut bacteria metabolites are involved in regulation of intraocular pressure or development of glaucoma. However, clinical data are lacking. Here, we wanted to compare level of trimethylamine (TMA), an uremic toxin produced by gut bacteria, along with betaine and trimethylamine N-oxide (TMAO), a substrate and a product of its metabolism, in the aqueous humor and in plasma of patients with glaucoma and their controls. METHODS: Twenty patients were selected for cataract phacoemulsification, and 20 patients selected for phacotrabeculectomy were enrolled in the study. Patients were matched with controls on systemic diseases and estimated glomerular filtration rate. Blood samples were collected in the preoperative suite, whereas aqueous humor samples were collected as the first step of both procedures. Subsequently, level of betaine, TMA and TMAO was analyzed by means of chromatography. RESULTS: In the aqueous humor, level of TMA, but not betaine or TMAO, was significantly higher in the phacotrabeculectomy group than in the phacoemulsification group. Plasma level of betaine, TMA and TMAO was similar between groups. In both groups, level of betaine and TMA, but not TMAO, was significantly higher in plasma than in the aqueous humor. CONCLUSION: TMA, but not TMAO or betaine level, is increased in the aqueous humor of patients with glaucoma. TMA might play a role in pathogenesis of glaucoma; however, prospective studies are needed to confirm our findings.

Occurrence of viral DNA in paired samples of corneal rim and cornea preservation fluid.

Corneal transplants have one of the highest success rates among all transplantological procedures. Corneas intended for transplantation are stored in a preservation fluid, which is then tested for bacterial and fungal infections. Among all analyses of infectious complications following corneal transplants, infections caused by bacteria or fungi are the most prominent. Surprisingly, however, apart from a few publications, there is a lack of data regarding the occurrence of viruses in donor corneas and the risk of transmitting these to their recipients. The intention of this research was therefore to determine the frequency with which human herpesvirus 1 (HHV-1), human herpesvirus 2 (HHV-2), and human adenovirus (HAdV) occur in transplanted corneal tissue, as well as in samples of preservation fluid. The study comprised 57 paired samples, with each pair consisting of a fragment of the corneal tissue remaining after its trepanation for transplantation surgery and a sample of corneal preservation fluid. Sample pairs were all tested for the presence of the DNA of three viruses (HHV-1, HHV-2, and HAdV) using real time PCR technique. Viral DNA was found in three of the tested corneas-HHV-1 DNA in one paired sample (1.8%) and adenovirus DNA in two single samples (3.5%). We postulate that virological testing of corneas for transplantation should be considered, particularly in the case of donors with increased risk factors for herpesvirus and adenovirus reactivation. J. Med. Virol. 89:732-736, 2017. © 2016 Wiley Periodicals, Inc.

Polymorphism of the LIG3 gene in keratoconus and Fuchs endothelial corneal dystrophy.

The product of the LIG3 gene encodes DNA ligase III, which is involved in the repair of oxidatively damaged DNA in the base excision repair pathway. We hypothesized that polymorphism in this gene may change susceptibility to oxidative stress and predispose individuals to the development of keratoconus (KC) and Fuchs endothelial corneal dystrophy (FECD). Therefore, we investigated the association between genotypes and haplotypes of the g.29661G>A polymorphism (rs1003918) and the g.29059C>T polymorphism (rs1052536) of the LIG3 gene and the occurrence of KC and FECD in patients with FECD (258 individuals) or KC (283) and ethnically matched controls (300). The A/A genotype and the A allele of the g.29661G>A polymorphism were associated with increased occurrence of KC, while the G allele of this polymorphism was positively correlated with a decreased occurrence of this disease. The T/C genotype of the g.29059C>T polymorphism was associated with decreased FECD occurrence. In addition, the AT haplotype was associated with increased occurrence of KC and FECD, while the GT haplotype was associated with decreased occurrence of these diseases. The g.29661G>A and g.29059C>T polymorphisms may play a role in the KC and FECD pathogenesis and can be considered as markers in these diseases.

Acinetobacter junii as an aetiological agent of corneal ulcer.

Rods of the Acinetobacter genus are present mainly in the external environment (e.g. water, soil) and in animals, while in humans they may comprise physiological flora. The main pathogenic species is Acinetobacter baumannii complex, which constitutes a common cause of nosocomial infections, particularly in patients with underlying diseases and risk factors (e.g. prior broad-spectrum antibiotic therapy, malignancy, central venous catheter, mechanical ventilation); however, infections of the eye caused by strains of Acinetobacter spp. are very rare. We report a unique case of community-acquired corneal ulcer caused by Acinetobacter non-baumannii (possibly A. junii), in a patient with no risk factors identified. The case highlights the need for obtaining a sample from the cornea for bacteriological culture in the case of suspected ophthalmic infection as identification of the pathogen, and assessment of its susceptibility profile enables proper antibiotic therapy, improves the outcome and may constitute an eyesight-saving management.

Mutagenesis of mitochondrial DNA in Fuchs endothelial corneal dystrophy.

Fuchs endothelial corneal dystrophy (FECD) is an age-related, slowly progressive disease, which may lead to loss of vision resulting from apoptosis of corneal endothelial (CE) cells, dysfunction of Descemet membrane (DM) and corneal edema. A growing body of evidence suggests that oxidative stress may play a major role in the pathogenesis of FECD and that mitochondria of CE cells are its main target. Mitochondrial DNA (mtDNA) is particularly prone to oxidative stress and changes in mtDNA were reported in FECD patients. In the present work we studied mtDNA damage and repair, mtDNA copy number, and the 4977bp common deletion in mtDNA in DM cells and peripheral blood lymphocytes (PBLs) isolated from FECD patients. PBLs from 35 FECD patients and 32 controls were challenged for 10min with hydrogen peroxide at 20µM and then left in a fresh medium for 3h, resulting in a decrease in mtDNA copy number in both groups. Damage to mtDNA was not fully repaired after 3h and the extent of remaining lesions was significantly higher in the patients than the controls. We observed a higher copy number and an increased extent of mtDNA damage as well as a higher ratio of the common 4977bp deletion in DM cells of FECD patients than the controls. Our results confirm that mutagenesis of mtDNA may be involved in FECD pathogenesis and disturbance in mtDNA sensitivity to damaging agent as well as changes in mtDNA damage repair along with alternations in mtDNA copy number may underline this involvement.

The relationship of TP53 and GRIN2B gene polymorphisms with risk of occurrence and progression of primary open-angle glaucoma in a Polish population.

INTRODUCTION: Glaucoma is characterized by optic neuropathy of the retinal ganglion cells (RGCs). Retinal ganglian cell death may be mediated by apoptosis. TP53 is involved in this process. It can also be found that excitotoxicity contributes to apoptosis by excess stimulation of glutamate receptors. The aim of this study was to evaluate the relationship of the TP53 (rs1042522) and GRIN2B (rs3764028) gene polymorphisms with risk of occurrence of primary open-angle glaucoma (POAG). MATERIAL AND METHODS: The study population consisted of 186 patients and 188 healthy subjects. Genomic DNA was extracted from peripheral blood. Analysis of the gene polymorphisms was performed using PCR-RFLP. RESULTS: Comparison of the distributions of genotypes and alleles of the rs1042522 and rs3764028 polymorphisms showed no statistically significant differences between POAG patients and controls (p > 0.05). There was a statistically significant association of the rs1042522 polymorphism with progression of POAG depending on the retinal nerve fiber layer (p = 0.019). However, no significant differences between rs3764028 polymorphism and clinical parameters of POAG were observed (p > 0.05). CONCLUSIONS: The TP53 Arg72Pro and GRIN2B -421C/A gene polymorphisms were not associated with risk of occurrence of POAG in the Polish population. However, the Arg72Pro polymorphism of the TP53 gene may be related to progression of POAG.

The role of the 148 Asp/Glu polymorphism of the APE1 gene in the development and progression of primary open angle glaucoma development in the Polish population.

Glaucoma is an ocular disorder that is characterized by progressive degeneration of the optic nerve and loss of visual field (VF). Recent data have suggested that the level of oxidative DNA damage in human trabecular meshwork is significantly increased in glaucomatous patients as compared to controls. It was also noted that progressive loss of visual field may by connected with elevated levels of oxidative DNA lesions. This hypothesis may suggest the role of an inefficient base excision repair pathway in primary open angle glaucoma (POAG) pathogenesis. The aim of the study was to evaluate the association of the 148 Asp/Glu APE1 gene polymorphism with the risk of POAG development. One hundred fifty patients with POAG and 190 controls were enrolled in our study. Gene polymorphisms were analyzed by PCR-CTPP. We did not observe a statistically significant difference between the frequencies of alleles and genotypes of the 148 Asp/Glu APE1 gene polymorphism in POAG patients and controls. However, the presented study indicated that 148 Asp/Glu of the APE1 gene was associated with decreased risk of progression of POAG with reference to the parameter VF. We suggest that the 148 Asp/Glu APE1 gene polymorphism may decrease the risk of POAG progression.

Clinical and microbiological efficacy of levofloxacin administered three times a day for the treatment of bacterial conjunctivitis.

PURPOSE: This randomized, investigator-masked study aimed to compare the clinical and microbiological effectiveness of three times daily administration of levofloxacin 0.5% eyedrops with the classic, more frequent dosing in patients with bacterial conjunctivitis. METHODS: A total of 120 patients with symptoms of bacterial conjunctivitis were enrolled. The patients were randomly assigned to receive 0.5% levofloxacin eyedrops three times daily to each eye for 5 days (experimental dosage group), or every 2 hours on days 1 and 2, and then every 4 hours on days 3-5 (up to four times per day) (classic dosage group). Ocular symptoms and signs were assessed on day 1, days 3 to 4, and 7 +/- 1 visits. Conjunctival cultures were obtained at baseline and final visits. Clinical outcomes were based on resolution of cardinal signs. Microbial outcomes were based on culture results. RESULTS: Eighty-six patients (41 experimental dosage, 45 classic dosage) were evaluated. There was no difference between the groups in frequency of patients with clinical outcome resolved (85.4% in experimental vs 93.3% in classic dosage group, p=0.3). The microbial eradication rates did not differ statistically between the groups (92.7% vs 95.6%, respectively, p=0.67). CONCLUSIONS: There was no statistically significant difference in the efficacy or safety between the two methods of drug administration. Analysis of the results of compliance supported our conclusion that the less frequent method of dosing of 0.5% levofloxacin eyedrops was more convenient for patients and resulted in better adherence to the drug-dosing scheme.

In vivo confocal microscopy of corneal grafts shortly after penetrating keratoplasty.

PURPOSE: To describe the microstructural status of corneal grafts shortly after penetrating keratoplasty (PK) and to evaluate the efficacy and safety of confocal microscopy in examining corneal grafts at that time. METHODS: A confocal microscope with a 40 x front lens was used to examine corneal grafts in 32 patients (32 eyes) 4 days after PK. Images were analyzed, and endothelial cell density counts were compared with presurgical, eye bank values determined by specular microscopy. RESULTS: Microstructural alterations of the graft included epithelial and stromal edema, epithelial degeneration in both superficial and basal cell layers, dark stromal striae, activated keratocytes, and needle-like structures in the stroma. Descemet membrane folds were visible in 31 of 32 grafts; in 1 graft, the dense stromal edema did not allow imaging of posterior layers. Stromal nerve fibers were imaged in 28 grafts (88%). Endothelial cell density ranged from 1666 to 2548 cells/mm2 (mean+/-SD, 2125+/-283 cells/mm2); perioperative endothelial cell density loss varied from 0% to 29% (mean, 12%). No adverse reactions or signs of worsening of clinical condition were observed after the examination. CONCLUSIONS: White light scanning slit confocal microscopy permits imaging of a graft's microstructure (including epithelium and stromal layers), as well as calculation of endothelium cell density, as soon as 4 days after PK. The most frequently observed morphologic alterations of corneal grafts shortly after PK include epithelial and stromal edema, epithelial degeneration, stromal striae, and Descemet membrane folds. Stromal nerves can still be seen in the graft 4 days after PK.

The efficacy and safety of timolol maleate versus brinzolamide each given twice daily added to travoprost in patients with ocular hypertension or primary open-angle glaucoma.

PURPOSE: To compare the efficacy and safety of timolol maleate 0.5% versus brinzolamide 1% when added to travoprost 0.004% in patients with ocular hypertension or primary open-angle glaucoma. DESIGN: A prospective, double-masked, randomized, active-controlled, parallel comparison. METHODS: Qualified patients at Visit 1 were placed on travoprost dosed every evening for 4 weeks and then were randomized at baseline (Visit 2) to the addition of timolol maleate or brinzolamide each given twice daily. Patients returned to clinic at Week 4 (Visit 3) for a safety visit and Week 12 (Visit 4) for an efficacy visit. At Visits 2 and 4 the intraocular pressure (IOP) was measured at 08:00, 12:00, and 16:00 hours. RESULTS: Ninety-seven patients on brinzolamide had a baseline diurnal IOP of 21.5+/-2.2 mmHg and 95 on timolol maleate had 21.3+/-2.5 mmHg, each added to travoprost. The diurnal mean IOP at Week 12 was 18.1+/-2.7 mmHg for brinzolamide and 18.1+/-3.0 mmHg for timolol maleate (p=0.96). There was no statistical difference found between treatment groups in the absolute level of pressure, or in the reduction in IOP from baseline, at each time point or for the diurnal curve (p>0.05). There was no significant difference for any adverse event between groups (p>0.05), with the most common side effect being conjunctival hyperemia in 15/97 (16%) brinzolamide and 6/95 (6%) timolol treated patients (p=0.06). CONCLUSIONS: This study showed that brinzolamide provides similar safety and efficacy compared to timolol maleate when added to travoprost.

In vitro angiomodulatory activity of sera from type 2 diabetic patients with background retinopathy.

Diabetic retinopathy is the leading cause of adult vision loss and blindness. The most important contributors to the development of diabetic retinopathy are hyperglycemia and hypoxemia that lead to increased vasopermeability, endothelial cell proliferation, and pathological neovascularization. In our previous studies, close relationship between proangiogenic activity of sera from type 2 diabetes mellitus patients (DM2) with background retinopathy, assessed in the in vivo serum-induced mouse cutaneous test (SIA), and VEGF and IL-18 serum concentration were observed. Moreover, it was clearly shown that IGF-1 might play an important role in the negative regulation of neoangiogenesis induced by DM2 patients' sera by diminishing the VEGF stimulatory effect. To confirm the observed phenomenon we evaluated the effect of DM2 patients' sera on the in vitro proliferative activity of human endothelial cells, which is critical for the sprouting and generation of new blood capillaries. Endothelial proliferative activity was significantly higher in the presence of sera from DM2 patients than from healthy controls (P<0.001), as estimated by the MTT test. Moreover, the examined sera from DM2 patients were characterized by increased IL-18 (P<0.05), diminished IGF-1 (P<0.02), and unchanged VEGF levels compared with those in controls. In conclusion, the present study showed a strong stimulatory effect of DM2 patients' sera on the proliferation of endothelial cells, which, along with the findings of our previous studies, proves that the described phenomenon is universal and valid for both animal and human endothelium.

Culture of the primary corneal epithelium as a potential component of test batteries for eye irritancy testing.

One of the main goals for toxicologists working on the development of in vitro tests is to replace the animal-based eye irritation test. Inflammation is one of the mechanisms which have not been covered sufficiently by the existing in vitro ocular irritancy test systems. As there are major species differences between the human and rabbit eye inflammation mechanisms, the most relevant test system is the human eye itself. The current study focused on an evaluation of the practical availability of human corneal epithelial cells for routine eye irritancy testing. Human corneal epithelium cell cultures were used to assess the effects of lipopolysaccharide on IL-1 beta release. The findings indicated that cytokine release can be augmented by the presence of the complement system, which is normally found in tears. However, the corneal cells were found to be highly resistant to the complement system, which can be attributed to the very high expression of CD59, a powerful complement regulatory protein found in the corneal epithelium. It is estimated that discarded corneas from tissue banks could provide enough material for routine testing by this method.

The importance of various factors relating to the morphological quality of corneas used for PKP by the Warsaw Eye Bank from 1996 to 2002.

To examine the effect of various factors such as: donor's age, cause of death, time between death and preservation and duration of preservation on the morphological quality of corneas used for Penetrating Keratoplasty (PKP). Our purpose was to assess the role of the above factors influencing the corneal overall rating and endothelial cell density. The present data regarding the donor's age and time between death and preservation of corneas obtained from eye banks belonging to the European Eye Bank Association and Polish eye banks were compared. Corneoscleral buttons and data concerning donors were obtained by eye bank technicians and collected in Warsaw Eye Bank in the years 1996-2002. The quality of the corneas recovered was evaluated by means of slit lamp Nikon NS-IV and specular microscope KONAN. Cardiorespiratory failure and cardiac arrest were the most frequent cause of the donors' death. In many cases donors with confirmed brain death who were given the corneas, were also donors of internal organs like heart, kidneys, pancreas and liver. The number of donors with disseminated neoplasm disease--complete disadvantage for removing corneas increased during the eye bank activity. The epithelial and endothelial layers sometimes underwent mechanical trauma in the group of donors because of sudden death. From 1996 to 2002 more than 50% of the donors were over 60 years of age. There were many problems with receiving corneas from younger donors. The overall rating tissues which were obtained in a very short time after death (to 5 hours) was higher (Excellent and Very Good) compared with corneas removed 8-12 hours after the donor's death. The increasing percentage of endothelial cell loss was observed in all the corneas after about 7 days of preservation in the medium. The mean endothelial cell density slightly decreased with donor's age, but its suggested range of the factors possibility of finding the corneas with high number of endothelial cell density both in younger and older donors. The average time from death to preservation was similar in eye banks preserving corneas at +4 degrees C (8-10 hours) compared with eye banks using mainly the organ culture method. The rating of the morphological state of corneas suitable for PKP depends on the time between death and preservation, donor's age, cause of death and time of preservation of corneas. Corneas obtained shortly after the donor's death showed a higher endothelial cell density and better overall rating than those removed after relatively longer period after the donor's death. An increasing percentage of endothelial cell loss was observed after 7 days of preservation independent of other factors.

[The attempt to identify mutations in TIGR gene in Polish patients with primary open angle glaucoma]. / Próba identyfikacji mutacji w genie TIGR w polskiej populacji chorych na jaskre pierwotna otwartego kata.

PURPOSE: The case work presents studies on the identification of the most frequent TIGR mutations in Polish population with primary open angle glaucoma. The TIGR gene was identified in a GLC1A locus on chromosome 1 (1q) in a family with juvenile primary open angle glaucoma. The gene encodes TIGR protein (trabecular meshwork inducible gluco-corticoid response protein)--trabecular meshwork glucoprotein. MATERIAL AND METHODS: Ophthalmologic examination was performed in twenty subjects with juvenile primary open angle glaucoma. The blood samples were taken for DNA analyses. RESULTS: Neither any mutations nor polymorphic changes in TIGR gene were found. CONCLUSION: Our studies have not identified any mutations in exon 3 of TIGR gene. We cannot exclude, however, that mutations are localised in other exons or regulatory region of examined gene. The questions how many genes, how many mutations of these genes and how often they contribute to glaucoma in general population are still open? These are important questions to answer in order to get closer to understanding extremely complicated aetiology of glaucoma.

[Genetic studies on primary open angle glaucoma]. / Badania genetyczne rodzin w jaskrze pierwotnej otwartego kata.

Case work presents the newest studies on molecular genetics in primary open angle glaucoma. The molecular genetics in all types of glaucoma have been a subject of investigations for the last few years. As a result, two loci (GLC3A and GLC3B) have been identified for primary congenital glaucoma, one locus (GLC1A) for juvenile primary open angle glaucoma and further two loci (GLC1B and GLC1C) for adult-onset primary open angle glaucoma. The gene TIGR (trabecular meshwork inducible glucocorticoid response protein) localised in GLC1A was described last year for the first time, thereafter there were trials on mutations within this gene conducted successfully. In this review there are studies presenting the mapping of the first five GLC loci and identification of mutations.

[The history of correction of refractive errors: refractive surgery]. / Historia korekcji wad refrakcji - chirurgia refrakcyjna.

The evolution of particular methods of refractive surgery is presented including the origins of radial keratotomy, relaxing incisions, keartophakia, epiferatophakia and keratomileusis as well as recent adaptation of lasers for the surgical treatment of refractive error. Such famous persons as Barraquer, Sato, and Fidorov are discussed in particular. The history not only of great achievements but also of some of the false ideas which accompanied efforts to obtain the emmetropia is presented.

[Deep sclerectomy ab externo with implant: description of surgery technique]. / Gleboka sklerektomia ab externo z implantem--opis techniki operacyjnej.

The authors present the modified associated technique of deep sclerectomy ab externo with implant. The indications, advantages and disadvantages are presented.

[Corneal wound healing after penetrating keratoplasty with EGF application. Experimental studies]. / Obserwacje gojenia sie rany rogówki oka po przeszczepach drazacych z zastosowaniem naskórkowego czynnika wzrostu (EGF). Badania doswiadczalne.

PURPOSE: To check post penetrating keratoplasty (PK) corneal wound healing characteristics after epidermal growth factor (EGF) application and to compare it with controls. MATERIAL AND METHODS: The PK was performed in the group of 72 young, healthy New Zealand rabbits (36 females and 36 males). Slit-lamp examination, tonometry and corneal topography by Tomey's corneal modeling system (TMS-1) were carried out before and after surgery. The PK was performed in both eyes. Half of animals were used as a bilateral donor for the other half, with a rule: right eye cornea to the right eye and left eye cornea to the left eye. As a result, after completed surgery 36 rabbits had bilateral grafts. The animals were divided into 3 equal groups (12 in each). Two drops of the human recombined EGF dissolved in the saline solution with concentration varied from 500 to 1500 ng in each drop were applied to the right eye according to schedule. The left eye was used as a control and did not receive EGF. Time of observation varied from 24 hours to 6 months. The tensinometry and the histopathologic study--light and electron microscopy were performed to determine corneal scarring. RESULTS: The wound healing pattern after PK was characteristic and constant in each group. The corneal wound healing significantly accelerated in the EGF treated group of rabbits compared with the controls (p < 0.05). In the group of rabbits receiving 1000 ng of hrEGF 3 times/day, after two weeks of application we noted increase of the wound strength up to 600 folds, comparing with controls. Well-organized scar was histologically seen on the 21st post-surgery day. The post-operative corneal astigmatism was less expressed in the eyes treated with EGF comparing to controls. CONCLUSIONS: These preliminary results of our experimental study indicated accelerated effect on the corneal wound healing after PK with topical, low dose hrEGF application. Clinical observation of utilization of similar low doses of the hrEGF after PK--is in progress.