Safety of an Intracameral Fixed Combination for Mydriasis and Intraocular Anaesthesia During Cataract Surgery.

Purpose: To compare the safety of a standardized, commercially available intracameral combination of mydriatics and anesthetic (ICMA) with a reference topical mydriatic regimen for cataract surgery. Patients and Methods: The safety results from two international, randomized, controlled clinical studies were combined to compare ICMA at the beginning of cataract surgery (ICMA group) to the reference topical mydriatic regimen (reference group). Data were collected on ocular and systemic adverse events, corneal and anterior chamber examination, endothelial cell density, retinal thickness and visual acuity. Analysis was performed on a pooled safety set from both studies, preoperatively and up to 1 month postoperatively. Results: 342 patients received ICMA and 318 the reference topical regimen. Ocular adverse events were reported in 17.0% of patients in the ICMA group and 18.6% in the reference group. No difference was shown between groups in endothelial cell density (2208 ± 498 cells/mm2 for ICMA group versus 2241 ± 513 cells/mm2 for the reference group; p=0.547) and retinal thickness (change from baseline less than 50 µm in 94.7% versus 95.0% of patients, respectively) at 1 month postoperatively. At 1-day post-surgery, less patients in the ICMA group had moderate or severe (Grades 2 and 3) superficial punctate corneal staining (3.9% versus 7.0% for the reference group; p=0.064). Postoperatively, some ocular symptoms were also less frequently reported in the ICMA group. Best-corrected visual acuity increased in 96.0% of patients in the ICMA group and 95.8% in the reference group at 1 month. Conclusion: ICMA injection at the beginning of cataract surgery was demonstrated to be safe and may also provide perioperative and postoperative advantages over the standard topical mydriatic regimen.

Successive Acanthamoeba Corneal Isolates Identified in Poland Monitored in Terms of In Vitro Dynamics.

BACKGROUND: Amoebae of the genus Acanthamoeba cause a sight-threatening infection called Acanthamoeba keratitis. It is considered a rare disease in humans but poses an increasing threat to public health worldwide, including in Poland. We present successive isolates from serious keratitis preliminary examined in terms of the identification and monitoring of, among others, the in vitro dynamics of the detected strains. METHODS: Clinical and combined laboratory methods were applied; causative agents of the keratitis were identified at the cellular and molecular levels; isolates were cultivated in an axenic liquid medium and regularly monitored. RESULTS: In a phase-contrast microscope, Acanthamoeba sp. cysts and live trophozoites from corneal samples and in vitro cultures were assessed on the cellular level. Some isolates that were tested at the molecular level were found to correspond to A. mauritanensis, A. culbertsoni, A. castellanii, genotype T4. There was variability in the amoebic strain dynamics; high viability was expressed as trofozoites' long duration ability to intense multiply. CONCLUSIONS: Some strains from keratitis under diagnosis verification and dynamics assessment showed enough adaptive capability to grow in an axenic medium, allowing them to exhibit significant thermal tolerance. In vitro monitoring that was suitable for verifying in vivo examinations, in particular, was useful to detect the strong viability and pathogenic potential of successive Acanthamoeba strains with a long duration of high dynamics.

Pupil diameter during cataract surgery after intracameral injection of the first ready-to-use combination of mydriatics and anaesthetic at the beginning of surgery in patients with a preoperative pupil diameter <6 mm.

PURPOSE: We evaluated, in a real-life setting, the effect of Mydrane® (ready-to-use combination of tropicamide, phenylephrine hydrochloride and lidocaine, injected into the anterior chamber at the beginning of cataract surgery to induce mydriasis and intraocular anaesthesia) on the pupil diameter during cataract surgery in patients with a preoperative pupil diameter <6 mm after the use of topical mydriatics. METHODS: We collected and analysed the data of 59 consecutive patients whose pupils dilated to a diameter <6 mm after the administration of mydriatic eye drops during the preoperative visit and who received Mydrane® during cataract surgery. RESULTS: In the group of 59 patients with a preoperative pupil diameter <6 mm after topical mydriatics, cataract surgery was performed in 36 patients (61.0%) using only Mydrane® to obtain mydriasis, with no additional drug or medical device. The mean pupil diameters in this group (36 of 59) during the preoperative assessment after topical mydriatics and just before capsulorhexis when Mydrane® was injected during surgery were 5.1 ± 0.74 and 6.15 ± 1.14 mm. Additional drugs were used in 23 patients (39%). In this group, the mean pupil diameters after topical mydriatics and just before capsulorhexis using Mydrane® were 4.58 ± 1.06 and 5.6 ± 1.26 mm, respectively. CONCLUSION: In a real-life setting, the mean pupil diameter achieved during cataract surgery after the intracameral injection of Mydrane® in patients with a preoperative pupil diameter <6 mm was over 1 mm larger than the mean pupil diameter after topical mydriatics, despite the trauma caused by the operation.

ASSOCIATED FACTORS AND SURGICAL OUTCOMES OF MICROCYSTOID MACULAR EDEMA AND CONE BOUQUET ABNORMALITIES IN EYES WITH EPIRETINAL MEMBRANE.

PURPOSE: To investigate the associated features and the surgical outcomes in eyes with microcystoid macular edema (MME) and cone bouquet abnormalities (CBA) undergoing epiretinal membrane peeling. METHODS: Retrospective study including patients who underwent pars plana vitrectomy for idiopathic epiretinal membrane. Factors associated with MME and CBA at baseline and their regression were identified with logistic regression models. Postoperative visual acuity was evaluated with linear mixed models from baseline to 12 months. Risk factors for new or worsened macular edema were explored with a Cox regression model. RESULTS: One hundred and eighty-seven eyes are included in this study; 30 eyes (16%) had MME and 53 eyes (28%) had CBA preoperatively. Microcystoid macular edema was associated with severe epiretinal membrane stage (odds ratio [95% confidence interval] = 3.6 [1.3-12.7], P = 0.02); CBA was inversely associated with ectopic inner foveal layer thickness (odds ratio [95% confidence interval] = 0.97 [0.97-0.99] for each 1- µ m EIFL increase, P = 0.006). Eyes with MME had worse visual acuity after epiretinal membrane peeling ( P = 0.01) and were at risk of macular edema worsening (hazard ratio [95% confidence interval] = 2.22 [1.01-5.16], P = 0.04). Older age was associated with MME persistence (odds ratio [95% confidence interval] = 2.46 [1.06-6.82] for each 10-year increase, P = 0.04). No significant associations were found for CBA. CONCLUSION: Although CBA had no prognostic consequences, MME was associated with suboptimal visual recovery and less efficient control of inflammation after surgery. Degeneration of Müller cells may have an alleged role, and further imaging and functional tests are warranted.

Canaloplasty - Efficacy and Safety in an 18-Month Follow Up Period, and Analysis of Outcomes in Primary Open Angle Glaucoma Pigmentary Glaucoma and Pseudoexfoliative Glaucoma.

PURPOSE: Open-angle glaucoma (OAG), accounting for 90% of all glaucoma cases, is a progressive optic nerve neuropathy. It may lead to irreversible loss of visual field and complete blindness. When conservative treatment becomes insufficient to stop OAG progression, a surgical intervention is considered. Currently, canaloplasty procedure is being introduced instead of conventional trabeculectomy for invasive OAG treatment. The aim of the study is to asses safety and efficacy of canaloplasty. METHODS: This prospective study included 67 eyes that received 360° canaloplasty with placement of a tensioning suture. Primary OAG (n = 35), secondary OAG in pseudoexfoliative syndrome (n = 13), and pigmentary glaucoma (n = 19) patients were included. Control check-ups were conducted pre-operatively and in a 18-month follow-up time. Study endpoints involved reduction in IOP values and in the number of glaucoma medications after the intervention. RESULTS: The intervention led to a significant 38% reduction in IOP value from the preoperative baseline to 18 months after the intervention. The number of medications decreased significantly by 89%. At 18 months postoperative, 79% eyes did not require any glaucoma medications. The incidence of complications after canaloplasty was low, and none of the adverse effects were vision threatening. A surgically-induced astigmatism was the most frequent complication. Pigmentary glaucoma patients were the most beneficial subgroup, with 50% reduction in IOP, the highest success rate, and 98% reduction in the number of medications used. CONCLUSION: This study proved that canaloplasty is an efficient and safe procedure in OAG eyes.

First Report of a Case of Ocular Infection Caused by Purpureocillium lilacinum in Poland.

This report describes the first case of an ocular infection induced by Purpureocillium lilacinum in Poland. The patient was a 51-year-old immunocompetent contact lens user who suffered from subacute keratitis and progressive granulomatous uveitis. He underwent penetrating keratoplasty for corneal perforation, followed by cataract surgery due to rapid uveitic cataract. A few weeks later, intraocular lens removal and pars plana vitrectomy were necessary due to endophthalmitis. The patient was treated with topical, systemic, and intravitreal voriconazole with improvement; however, the visual outcome was poor. The pathogen was identified by MALDI-TOF MS.

Neuroprotective Factors of the Retina and Their Role in Promoting Survival of Retinal Ganglion Cells: A Review.

Retinal ganglion cells (RGCs) play a crucial role in the visual pathway. As their axons form the optic nerve, apoptosis of these cells causes neurodegenerative vision loss. RGC death could be triggered by increased intraocular pressure, advanced glycation end products, or mitochondrial dysfunction. In this review, we summarize the role of some neuroprotective factors in RGC injury: ciliary neurotrophic factor (CNTF), nerve growth factor (NGF), brain-derived neurotrophic factor, vascular endothelial growth factor, pigment epithelium-derived factor, glial cell line-derived neurotrophic factor, and Norrin. Each, in their own unique way, prevents RGC damage caused by glaucoma, ocular hypertension, ischemic neuropathy, and even oxygen-induced retinopathy. These factors are produced mainly by neurons, leukocytes, glial cells, and epithelial cells. Neuroprotective factors act via various signaling pathways, including JAK/STAT, MAPK, TrkA, and TrkB, which promotes RGC survival. Many attempts have been made to develop therapeutic strategies using these factors. There are ongoing clinical trials with CNTF and NGF, but they have not yet been accepted for clinical use.

Cowpox: How dangerous could it be for humans? Case report.

Cowpox is a rare zoonosis transmitted to humans mainly from cats. The disease usually causes skin lesions; however, the ocular form may lead to other serious complications. We describe a case of cowpox in a rare location of the upper eyelid of an immunocompetent male, which lead to necrosis of the upper eyelid, keratitis and leucomatous opacity, and the neovascularization of the cornea. The patient underwent several surgeries, including reconstruction surgery of the eyelids, correction of the medial canthus, and corneal neurotization with supraorbicular nerve transplantation. Suspicion of cowpox should be made in patients where there are poorly healing skin lesions accompanied by a painful black eschar with erythema and local lymphadenopathy. Ocular cowpox may lead to serious complications and possibly mimic anthrax. Diagnosis of cowpox can be confirmed by detection of cowpox virus DNA by polymerase chain reaction. Patients should be advised to protect themselves while handling sick animals.

Stable Mydriasis After Intracameral Injection of a Combination of Mydriatics and Anesthetic During Cataract Surgery: A Real-Life, Multicenter Study.

Purpose: To evaluate the effect of Mydrane (contains tropicamide, phenylephrine hydrochloride, and lidocaine hydrochloride) on time needed to induce mydriasis and mydriasis stability during cataract surgery. Methods: This was an observational, non-interventional, multicenter study of patients undergoing cataract surgery who received Mydrane for mydriasis and intraocular anesthesia. The study was conducted at seven ophthalmology departments at university hospitals in Poland. Patients admitted for cataract surgery within a 2-week period were asked to participate in the study. Patients whose pupils dilated to a diameter ≥6 mm after topical mydriatic administration during preoperative examinations were scheduled to receive Mydrane and included in the registry. No additional inclusion criteria were used. Patients' medical histories, examination results, and operative details were recorded. Pupil diameter was measured during surgery. Surgeons were asked to complete a Likert-based survey in parallel. Results: A total of 307 patients were enrolled. The mean pupil diameter was 7.0 ± 1.0 mm before capsulorhexis and 6.9 ± 1.2 mm before lens implementation. A pupil diameter ≥6 mm was achieved in 91.9% and 87.6% of patients before capsulorhexis and lens implantation, respectively. We asked 58 surgeons whether they agreed with the statement "Mydriasis was obtained in a short time after the administration of Mydrane"; the surgeons agreed with this statement after 92.2% (283/307) of surgeries. In addition, after 88.2% of surgeries, the surgeons agreed with the statement "Mydriasis was stable after the administration of Mydrane." Conclusions: Mydriasis was rapidly and stably obtained after Mydrane injection, as demonstrated by pupil diameter measurements during surgery and surgeons' feedback.

Immersion Biometry for Intraocular Lens Power Calculation with Fourth-Generation Formulas.

BACKGROUND: Fourth-generation formulas for intraocular lens power calculations, including the Barrett Universal II formula, the Olsen formula or the Holladay 2 formula, were thoroughly validated with optical biometry measurements. They precisely predict the effective lens position not only in normal eyes but also in eyes with unusual anatomy. However, in the setting of dense nuclear or posterior subcapsular cataracts, optical biometers fail to obtain accurate measurements and third-generation formulas, i.e. the Hoffer Q or the SRK/T, combined with ultrasound measurements are a method of choice. Considering that optical biometry was fine-tuned to immersion ultrasound, we hypothesize that fourth-generation formulas will yield precise intraocular lens power calculations with immersion ultrasound measurements. METHODS: We retrospectively analyzed 50 eyes of 50 patients who underwent uneventful cataract surgery. All patients had intraocular lens power calculated based on immersion ultrasound measurements. Refractive error predictions were compared between third-generation formulas and fourth-generation formulas. RESULTS: There were no statistically significant differences in the median absolute error between formulas. In the study, 86%, 88%, 86%, 84%, 88% and 80% of eyes were within 1 D of target refraction for the SRK/T, the Barrett II, the Hoffer Q, the Holladay 1, the Holladay 2 and the Olsen formula respectively. CONCLUSION: Fourth-generation formulas combined with immersion ultrasound produced similar results to third-generation formulas. However, the percentage of eyes within 1 D of target refraction remains inferior to previously reported results for optical biometry measurements.

Retinal arterial remodeling in patients with pheochromocytoma or paraganglioma and its reversibility following surgical treatment.

OBJECTIVE: Structural abnormalities in resistance arteries are a hallmark of patients with hypertension. In hypertensive patients with pheochromocytoma or paraganglioma (PPGL), it is still a matter of debate whether structural vascular changes are because of elevated blood pressure (BP) or to toxic effects of elevated circulating catecholamines. Hence, the aim of our study was to assess whether catecholamine excess and/or elevated BP affect the structure of small retinal arteries in patients with catecholamine-producing tumors. METHODS: The study included 27 patients with PPGL and 27 hypertensive patients. All patients underwent biochemical tests for catecholamine excess, echocardiography and analyses of scanning-laser-Doppler-flowmetry (SLDF) both at baseline and 12 months following surgical resection of PPGL. RESULTS: Baseline retinal arterial diameter, arterial wall thickness and wall cross sectional area (WCSA) were higher in patients with PPGL as compared with subjects without PPGL (arterial diameter: 110 ±â€Š16.5 vs. 99.5 ±â€Š10.8 µm, wall thickness: 16.3 ±â€Š6.0 vs. 13.5 ±â€Š4.0 µm, WCSA: 4953.9 ±â€Š2472.8 vs. 3784.1 ±â€Š1446.3 µm, P < 0.05). Significant correlations were noted between wall thickness and WCSA and echocardiographic parameters assessing diastolic and systolic function of left ventricle. No correlations between retinal parameters, BP level and plasma concentrations of metanephrines were observed. In patients with PPGL, there were postoperative decreases in wall thickness (16.4 ±â€Š15.8 vs. 14.8 ±â€Š4.7 µm; P = 0.011) and WLR (0.42 ±â€Š0.13 vs. 0.37 ±â€Š0.10; P = 0.003) at 12 months after surgical removal of tumors. CONCLUSION: This is the first study to demonstrate that catecholamine excess is related to thickening of retinal arteries independent of BP and reversible after surgical cure. These data support a role of catecholamines in vascular remodeling in PPGL patients.

Incidence and Characteristics of Endophthalmitis after Cataract Surgery in Poland, during 2010-2015.

Background: The assessment of the incidence and characteristic of acute and chronic postoperative endophthalmitis (POE) after cataract surgery in Poland during 2010-2015. Patients and methods: All hospitalizations of patients, in the National Database of Hospitalizations, who underwent cataract surgery alone or in combined procedures in Poland between January 2010 and December 2015, with a billing code of endophthalmitis, were selected. Acute endophthalmitis was identified if symptoms occurred within 1-42 days from the cataract surgery and chronic endophthalmitis if symptoms occurred ≥43 days after cataract surgery, respectively. Results: In total, 1331 cases of POE after 1,218,777 cataract extractions were identified. The overall incidence of POE decreased from 0.125% in 2010 to 0.066% in 2015. In multiple logistic regression analyses, increasing age was significantly associated with acute POE, while type II diabetes mellitus, extracapsular cataract extraction, and one-day surgery were significantly associated with chronic POE. The combined cataract surgery and male sex were significant risk factors for both acute and chronic POE. A total of 62.51% of all eyes affected by POE received antibiotic treatment and 37.49% had vitrectomy treatment. Conclusions: During the study period, the total incidence of postoperative endophthalmitis after cataract surgery decreased significantly.

Evaluation of in vitro effects of selected physical and chemical agents on detected in Poland Acanthamoeba strains ­ factors of increasing threats for public health

Acanthamoeba keratitis, the vision-threatening corneal disease reported with increasing frequency in Poland is difficult to treat due to extremely high resistance of the amoeba cysts to chemicals. The agents of possible anti-amoebic activity are still tested. Pathogenic Acanthamoeba samples/isolates acquired from severe cases of keratitis examined by molecular techniques to determine genotypes, compared to one another as well as to the environmental Acanthamoeba castellanii Neff strain were included in the studies. These strains were in vitro examined in terms of their sensitivity/resistance to selected chemicals and tolerance to temperature changes. Samples of the strains cultivated in vitro under bacteria-free conditions were monitored during different growth phases. Higher amoebic population dynamics was observed in both pathogenic Acanthamoeba strains during transfer to 37°C. Agents tested influenced population dynamics in different degree; they showed amoebostatic or amoebicidal effects, however a tendency toward induction of encystment also appeared. Because activation of the dormant cysts can lead to repeated development of amoebae, very important is cysticidal efficacy of chemicals. Further in vitro investigations on various Acanthamoeba strains with different chemicals are still necessary.

Blood pressure and glaucoma: At the crossroads between cardiology and ophthalmology.

Glaucoma is an optic nerve neuropathy of undetermined cause. Although many mechanisms are thought to be involved in the development and progression of the disease, only an increased intraocular pressure has been established as a clinically significant modifiable risk factor. Nevertheless, up to 40% of patients develop glaucoma without evidence of increased intraocular pressure. Ample evidence suggests that alterations in the control of arterial blood might negatively affect optic nerve function. However, evidence-based guidelines on the management of arterial blood pressure in glaucoma patients are lacking. Regrettably, intraocular pressure is generally not included as a secondary end-point in clinical trials on arterial hypertension. Considering the relative simplicity of intraocular pressure measurements and large number of patients included in hypertension studies, the benefits of including intraocular pressure as a secondary end-point could be of a great value for improving care for glaucoma patients. Therefore, closer collaboration between cardiologists and ophthalmologists is needed.

Effect of povidone iodine, chlorhexidine digluconate and toyocamycin on amphizoic amoebic strains, infectious agents of Acanthamoeba keratitis - a growing threat to human health worldwide.

INTRODUCTION: Free-living amoebae, ubiquitous in outer environments, in predisposing circumstances may exist as parasites, infectious agents of Acanthamoeba keratitis. In recent decades, the vision-threatening corneal infection is a growing human health threat worldwide, including Poland. The applied therapy is often ineffective due to diagnostic mistakes, various pathogenicity of Acanthamoeba strains and high resistance of cysts to drugs; many agents with possible anti-amoebic activity are still being tested. In the presented study, selected chemicals are investigated in terms of their in vitro effect on corneal and environmental Acanthamoeba strains. MATERIAL AND METHODS: Samples of a corneal isolate from a patient with severe Acanthamoeba keratitis,of assessed on the basis of genotype associations of 18S rRNA and the type strain, Acanthamoeba castellanii Neff cultivated in bacteria-free condition, were exposed to povidone iodine, chlorhexidine digluconate or toyocamycin. In vitro population dynamics of the strains were monitored and compared to those of control cultures. RESULTS: All chemicals showed anti-amoebic effects with different degrees of effectiveness. Significant differences were observed in the in vitro population dynamics, and the morpho-physiological status of A. castellanii Neff T4 and corneal strains determined as A. polyphaga T4 genotype, exposed to povidone iodine or toyocamycin, in comparison with chlorhexidine taken as reference. CONCLUSIONS: Time-dependent amoebstatic in vitro effects were demonstrated for all agents, in particular, the results of assays with povidone iodine are promising. No significant stimulation of encystation appeared; however, as cysticidal efficacy of chemicals is expected, complementary research is needed on different Acanthamoeba strains with modified agent concentrations and method application.

Comparison of chlorhexidine disinfectant in vitro effect on environmental and ocular Acanthamoeba strains, the amoebic agents of human keratitis − an emerging sight-threatening corneal disease in Poland

Small amoebae belonging to the Acanthamoeba genus complete their life cycles in different environmental niches as free-living protists however some of them are facultative parasites that can cause severe disease in humans. The sight-threatening Acanthamoeba keratitis develops in immune-competent persons, mainly in contact lens wearers; it is detected with increasing frequency along with the spread of contact lens use. The high abundance of the amoebae in the environment is important for dispersion and transmission of the infections among humans. Emerging threats for the public health generated by these amoebae is the serious medical problem worldwide. Nonspecific symptoms, similar to those occurring in the other eye diseases, diagnostic mistakes, the delay of an appropriate treatment, an exceptional high resistance of the amoebae to chemicals and drugs result in a prolonged course of the disease and often unsuccessful therapeutic management. Thus, different chemicals are still examined for their potential activity in vitro against various species, strains/isolates of Acanthamoeba. As the prolonged therapy often induces encystation subsequently leading to excystment and recurrences of amoebic keratitis, apart from anti-amoebic activity, cysticidal effect of examined agents is desirable. In the present study, results of our comparative investigations showed that cationic antiseptic chlorhexidine digluconate indicated in vitro anti-amoebic effect on environmental Acanthamoeba castellanii Neff strain and pathogenic corneal Acanthamoeba polyphaga T4 genotype. Amoebostatic effect of the disinfectant was expressed in reduced number of surviving amoebae in comparison to the respective control cultures; simultaneously, despite prolonged incubation with the agent no stimulation of encystation was noted. The corneal strain was more resistant to the tested compound than the Neff strain. The cysticidal efficacy of chemicals is very expected, thus further in vitro studies on pathogenic Acanthamoeba strains with different application chemicals pattern are needed.

Incidence and Characteristics of Cataract Surgery in Poland, during 2010-2015.

Background: To assess the incidence and characteristic of cataract surgery in Poland from 2010 to 2015 and to interpret these findings. Patients and methods: Data from all patients who underwent cataract surgery alone or in combined procedures in Poland between January 2010 and December 2015 were evaluated. Patient data were from the national database of hospitalizations maintained by National Health Fund. Data on the population of Poland were obtained from Central Statistical Office of Poland. Results: In total, 1,218,777 cataract extractions (alone or combined with other procedures) were performed in 1,081,345 patients during 2010-2015. Overall, the incidence of cataract surgery increased from 5.22/1000 person-years in 2010 to 6.17/1000 person-years in 2015. Phacoemulsification was performed in 97.46% of cataract extractions, and 3.02% of cataract extractions were combined procedures. The rate of one-day procedures increased from 28.3% in 2010 to 43.1% in 2015. The probability of second-eye surgery 12 months after the first-eye surgery increased from 44% in 2010 to 73% in 2015 (log-rank test p < 0.0001). Conclusion: In Poland, from 2010 to 2015, the total incidence of cataract surgery, the number of people who underwent surgery, and the number of one-day cataract surgeries increased significantly.

Amoebicidal or amoebostatic influence of disinfectants used in health facilities and laboratories on corneal strains of Acanthamoeba

Different Acanthamoeba species are amphizoic organisms distributed in wide range of habitats in natural and man-made environments; they are also detected on surfaces of equipment and accessories in health facilities. Some strains of the amoebae are causative agents of the vision-threatening human disease Acanthamoeba keratitis, mainly reported in contact lens wearers. An exceptional high resistance of Acanthamoeba trophozoites and particularly cysts to chemicals, disinfectants and drugs is believed as influencing difficulty resulting in unsuccessful therapeutic management. As Acanthamoeba keratitis is the serious medical problem worldwide, different chemicals with possible activity against environmental and clinical Acanthamoeba strains are tested. In our study, selected disinfectants used in health care settings and laboratories were tested and their efficacy against the corneal strains Acanthamoeba castellanii and A. polyphaga , and environmental A. castellanii Neff strain was assessed. Comparative assessment of results of the assays show that, apart from amoebistatic effects, the disinfectants indicated expected cysticidal efficacy.

In vivo confocal microscopy and in vitro culture techniques as tools for evaluation of severe Acanthamoeba keratitis incidents

Amphizoic amoebae belonging to the genus Acanthamoeba are known as etiological agents of sight-threatening Acanthamoeba keratitis. The leading risk factor for the development of this serious human disease is contact lens wearing which popularity increases worldwide, also in Poland. The disease with active epithelial inflammations, corneal ulcers, including loss of the visual acuity is a serious medical problem as an emerging threat for the public health related to improper contact lens hygiene. The treatment of the amoebic keratitis is difficult, often unsuccessful due to delayed proper diagnosis. The clinical picture of the disease, often with severe course is nonspecific, similar to that occurring in viral, fungal or bacterial keratitis, thus clinical symptoms alone are not sufficient to identify the causative agent of the amoebic infection. Early diagnosis is decisive for the suitable therapeutic management and the treatment efficacy. In our studies, several complicated, difficult to treat Acanthamoeba keratitis incidences pertaining Polish patients using contact lenses have been retrospectively analyzed in terms of the usefulness of non-invasive methods of in vivo confocal microscopy and in vitro culture techniques applied for diagnosis. Hyper-reflective double-walled spherical Acanthamoeba cysts, with a more reflective outer wall were detected in the epithelium and anterior layers of the corneal stroma. In vivo confocal microscopy, if available, may be a valuable, sensitive tool for diagnosis in late identified severe infections mainly with strong viability strains, however confoscan may offer limited value at lowintensity amoebic infections. The microscopic visualization of amoebae in slides prepared directly from corneal scraping and laboratory examinations of specimens from in vitro cultivated corneal isolates allow to confirm or verify results of in vivo examinations, furthermore to identify directly the pathogens and to clarify previous misdiagnoses.

Assessment of in vitro dynamics of pathogenic Acanthamoeba strains originating from contact lens wearers with infectious keratitis

Recently, incidents of Acanthamoeba keratitis, the vision-threatening eye disease, are reported with increasing frequency worldwide, particularly in contact lens wearers. In our study, the retrospective assessment of in vitro dynamics of subsequent pathogenic Acanthamoeba isolates cultured at 24°C, detected in Polish contact lens wearers with keratitis is presented and results compared with those of environmental A. castellanii Neff strain. There were delayed the proper diagnosis that influenced prolonged and severe course of this eye disease and treatment difficulties. The corneal material was examined directly to visualize developmental amoeba stages for diagnose verification, microbiologically tested for the specific identification of bacteriae and fungi, and in vitro grown in culture medium in temperature 24°C. Among twenty-six keratitis incidents analyzed, eleven were cases of Acanthamoeba keratitis; in the six of them, Acanthamoeba strains and concomitant bacterial and/or fungal infectious agents were detected. In vitro assays showed variability in population density of several clinical strains in the exponential growth phase expressed in various range of overall amoeba number and different proportion between trophozoites and cysts. The clear influence of temperature on the in vitro cultivation of the amoebae was observed: statistically significant lower population dynamics was revealed by most of pathogenic clinical isolates in comparison with those showed by environmental strain. The in vitro monitoring of dynamics of Acanthamoeba strains isolated from infected eyes may be helpful for diagnostics verification, especially in mixed infectious keratitis.