László Imre

Reliability and reproducibility of corneal endothelial image analysis by in vivo confocal microscopy

Abstract.Background: The aim of our study was to evaluate the reliability and reproducibility of in vivo slit-scanning corneal microscopy by endothelial analysis. Methods: We examined 12 normal eyes of 12 persons. We analysed intraobserver differences in group I and interobserver differences in group II by manual and automatic evaluation of endothelial images. We also compared the methods of analysis in the two groups. We observed endothelial density, mean cell area, coefficient of variation and number of cells within the frame. Results: In group I we found no significant difference between the observed parameters either by automatic or manual analysis. In group II no significant difference between the two observers was found using automatic analysis. On manual analysis, however, we observed a significant difference between endothelial density and the number of evaluated cells (P=0.0034 and P=0.0028). We observed a significant difference (P<0.001) between automatic and manual analysis concerning each parameter, with a rather tight linear correlation (Pearson correlation between 0.76 and 0.94). Conclusion: Automatic endothelial analysis was reliable and well reproducible in both – intraobserver and interobserver – groups. By manual evaluation, the clinical significance of interobserver differences can be disregarded. The differences between automatic and manual methods of analysis can be traced back to measurement technical reasons. We observed tight linear correlation between parameters. The data can be described well by linear regression. In vivo slit-scanning corneal microscopy may be an alternative to specular microscopic analyses for clinical use.

Confocal microscopic evidence of increased Langerhans cell activity after corneal metal foreign body removal.

Purpose The purpose of the study was to examine the corneal inflammatory reaction and Langerhans cells with confocal microscopy after metal foreign body removal. Methods Corneal metal foreign body was removed from 9 eyes of 9 consecutive patients 12.1±13.6 (4 to 72) hours after superficial angle grinder injury. Both eyes were examined with the Heidelberg Retina Tomograph II (HRT II) Rostock Cornea Module. Morphology and density of epithelium, nerves, metal deposits, keratocytes, endothelium, and Langerhans cells were compared to the uninjured fellow eyes (controls). Results Irregularity and partially missing superficial epithelium was found in all cases. Around the area of injury prolonged basal and wing epithelial cells were found in all eyes. The basal epithelium density is lower than in the control eye (p=0.043). Density of Langerhans cells (68.1±24.2/mm2) was increased in the epithelium, compared to controls (35.2±21.8/mm2, p=0.012). Keratocyte and endothelium densities were not different from that of controls. Some keratocytes showed signs of activation and the inhomogeneous background reflectivity revealed extracellular matrix alterations. Inflammatory reaction was observed up to 260 μm depth. The metal foreign body particles had high reflectivity and irregular edge. Conclusions In vivo confocal microscopy provided additional information to biomicroscopic signs such as epithelial damage and inflammation. It showed the effects of metal foreign bodies in the cornea: nerve damage and Langerhans cell density increase. Langerhans cells seem to play an important role in the inflammatory response after corneal foreign body injuries.

Drug reservoir function of human amniotic membrane.

PURPOSE The aim of the study was the quantitative pharmacokinetic evaluation of drug release from pretreated amniotic membrane (AM) in vitro. METHODS Cryopreserved AM pieces soaked in 3% ofloxacin ophthalmic solution were mounted in vertical Franz-diffusion cell system equipped with autosampler. In vitro release of ofloxacin was determined by quantitative absorbance measurement carried out with a UV spectrophotometer (wavelength 287 nm). Three groups were created according to the duration of soaking: 60 (Group 1), 120 (Group 2), and 180 (Group 3) minutes. Released amount of ofloxacin pro 1 cm(2) of AM (μg/cm(2)) was calculated in the period of 1 to 450 min. RESULTS Ofloxacin was detectable in the acceptor phase 1 min after mounting in all groups. Until 120 min, rapid increase of released ofloxacin could be observed. From 120 to 450 min, the amount of released ofloxacin showed a slower increasing pattern. Released ofloxacin in Group 1 was significantly lower than in Group 2 after 90 min (19.4±10.4 μg/cm(2), 51.6±20.7 μg/cm(2), respectively, P=0.044). In Group 3, cumulative drug release was higher than in Group at all timepoints. No significant difference could be demonstrated between Groups 2 and 3 at only 1 min timepoint. CONCLUSION Significant ofloxacin reservoir capacity of a single human amniotic layer could be demonstrated in vitro. AM acted as an ofloxacin slow release device for upto 7 h in vitro, depending on the duration of pretreatment of AM. Individual pretreatment of AM could increase beneficial effects of AM transplantation, especially in infectious keratitis.

Permeability of Human Amniotic Membrane to Ofloxacin In Vitro

PURPOSE The aim of this study was to develop a model to investigate the permeability of the amniotic membrane (AM) to ofloxacin eye drops, a widely used topical antibiotic in ocular surface disease after AM transplantation. METHODS AM pieces on cellulose acetate filter membranes were mounted in a vertical Franz-diffusion cell system equipped with an autosampler. In vitro release of 300 mg of 3% commercially available ofloxacin ophthalmic solution was determined by quantitative absorbance measurement carried out with a UV spectrophotometer (wavelength, 287 nm). Freshly prepared and cryopreserved AMs were compared. Filter membranes without AM served as positive controls. RESULTS Ofloxacin was detectable in the acceptor phase 1 minute after instillation, and a gradual increase of concentration could be detected in a period of 90 minutes in all groups. At 30 minutes 3.35% +/- 2.23% of ofloxacin penetrated the freshly prepared AM, 4.35% +/- 1.8% through cryopreserved AM compared with 17.52% +/- 3.91% filter membrane alone. At 90 minutes, penetration rates of ofloxacin were 5.04% +/- 1.11%, 5.26% +/- 3.21%, and 27.91% +/- 3.05%, respectively. Difference (P > 0.05; t-test) was not significant between freshly prepared and cryopreserved AMs. Compared with control, both membranes showed significant differences (P < 0.05; t-test) at all time points. CONCLUSIONS The in vitro model of the Franz-diffusion cell system was found to be applicable for drug permeability studies of human amniotic membranes to water-based solutions. The filter membrane and AM were permeable to a water-based solution of ofloxacin. Significant barrier function of the AM could be measured in ofloxacin permeability. Cryopreservation did not influence the permeability of the AM.

A Homogén Betegségcsoportok és a hozzá kapcsolódó finanszírozási rendszer elmúlt 25 éve

Absztrakt Magyarorszagon hosszas előkeszites utan, 1993-ban vezettek be az egeszsegugy finanszirozasanak uj rendszeret, amelynek egyik kozponti eleme a Homogen Betegsegcsoportok elven nyugvo modszer volt az aktiv fekvőbeteg-ellatas tekinteteben. A modszer egy atfogo rendszermodellbe illeszkedett, amelyet a Gyogyinfok szakemberei dolgoztak ki es publikaltak a rendszervaltas kornyeken. A vegul bevezetett rendszer sokban tukrozte ezt az elmeleti modellt, mai valtozata ugyanakkor tobb jelentős ponton is elter attol. A cikk celja ezen elteresi pontok azonositasa. Orv. Hetil., 2015, 156(29), 1155–1164.

Both Freshly Prepared and Frozen-Stored Amniotic Membrane Cells Express the Complement Inhibitor CD59

Amniotic membrane proved to be very effective tool in the treatment of a number of ocular surface diseases. The amniotic membrane, however, has to be stored before its transplantation onto the ocular surface followed by mandatory serologic control in order to exclude the transmission of certain viruses. Therefore it is most important to study if cryopreservation of the membrane affects cell surface expression of the molecules. We measured cell surface expression of CD59, a membrane-bound complement inhibitor on the cells of freshly prepared and cryopreserved amniotic membrane. Cells of amniotic membrane were separated mechanically. Epithelial and mesenchymal cells were identified by the intracellular expression of nanog and the cell surface ICAM1 positivity, respectively. Multicolor flow cytometric immunophenotyping was used for determination of the CD59 expression. CellQuest-Pro software program (Becton Dickinson) was used both for measurements and analysis. CD59-positive cells could be detected in all investigated samples and in all investigated cell types, although the expression level of CD59 differed. CD59 was expressed both on freshly prepared and frozen-stored samples. Higher level of CD59 was detected on ICAM1+ mesenchymal cells than on nanog+ epithelial cells. Our findings indicate that amniotic membranes maintain their complement inhibiting capacity after cryopreservation.

Specificity of in vivo confocal cornea microscopy in Acanthamoeba keratitis.

Purpose To report on the presence of 4 different structures visualized by confocal microscopy in patients whose clinical presentation suggested infection by Acanthamoeba. Methods Data and charts of 28 consecutive patients were analyzed in a retrospective study. Four types of structures were recognized by confocal microscopy performed with HRT II Rostock Cornea Module: trophozoites, double-walled cysts, signet rings, and bright spots. The 28 patients (mean age 30.8 years, range 17-61 years, 10 male, 18 female) were divided into 4 groups according to the diagnosis established later by microscopic examination of smear, culture, response to therapy, and the course of keratitis. The 4 groups were Acanthamoeba keratitis (AK), Acanthamoeba suspect (AK-suspect), bacterial keratitis (BK), and fungal keratitis (FK). Results The rate of patients in AK, AK-suspect, FK, and BK groups where bright spots were found were 100%, 100%, 40%, and 55%, respectively. The sensitivity of presence of bright spots in the in vivo confocal microscopy in Acanthamoeba keratitis was 100% (95% confidence interval [CI] 73.5% to 100.00%) and specificity was 50% (CI 24.7% to 75.4%). When cases where the only signs of Acanthamoeba were bright spots were excluded, and only those cases were counted where any of cysts, trophozoites, or signet rings were also found, the sensitivity was 67% (95% CI 34. 9% to 90.1%) and the specificity was 94% (95% CI 69.8% to 99.8%). Conclusions The relatively high rate of bright spots in non-Acanthamoeba keratitis challenges the assumption that bright spots seen by confocal microscopy are a specific indication of Acanthamoeba keratitis.

Az otthoni szakápolás egészségbiztosí tási vonatkozásainak elemzése Magyarországon

Bevezetes: Az otthoni szakapolas 1996 vegen indult meg Magyarorszagon. Celkitűzes: A szerzők celja a magyar otthoni szakapolasi rendszer egeszsegbiztositasi es igenybeveteli mutatoinak bemutatasa. Modszer: Az elemzesben szereplő adatok az Orszagos Egeszsegbiztositasi Penztar finanszirozasi adatbazisabol (2001–2012) szarmaznak. Elemeztek a betegszamokat, a vizitszamokat, a szakapolasi es szakiranyu terapias tevekenysegek (gyogytorna, fizioterapia, logopedia) aranyat. Eredmenyek: Az ellatott betegek szama a 2001. evi 36 560-rol 2012-re 51 647-re nőtt, 41,3%-kal emelkedett. A teljesitett vizitek szama ugyanezen időszak alatt 841 715-ről 1 194 670-re, 41,9%-kal nőtt. A szakapolasi es szakiranyu terapias tevekenysegek, valamint az apolasi fokozatok vonatkozasaban jelentős megyek kozotti eltereseket figyeltek meg. A szakapolasra kifizetett osszegek aranya a legmagasabb volt Nograd (80,4%), Szabolcs-Szatmar-Bereg (79,7%) es Komarom-Esztergom (74,6%) megyekben, mig a legalacsonyabb Zala (53,0%) es Csongrad (52,7%...INTRODUCTION Home nursing care was introduced in Hungary in 1996. AIM The aim of this study was to analyse health insurance data and utilization indicators of the Hungarian home nursing care. METHOD Data derived from the database of the National Health Insurance Fund Administration (2001-2012). The number of patients and visits, and the ratio of special nursing and special therapy (physiotherapy, speech therapy) were analysed. RESULTS The number of patients increased by 41.3% from 36.560 (2001) to 51.647 (2012). The number of visits also increased by 41.9% from 841.715 (2011) to 1.194.670 (2012). Significant geographical inequalities were found in the ratio of special nursing and special therapy as well as nursing needs. The ratio of reimbursement for special nursing was the highest in county Nógrád (80.4%), Szabolcs-Szatmár-Bereg (79.7%) and Komárom-Esztergom (74.6%), while the lowest in county Zala (53.0%), Csongrád (52.7%) and Budapest (47.9%). CONCLUSIONS There are significant inequalities in the home nursing care in Hungary. In order to decrease these inequalities, specific guidelines should be developed for home nursing care.

Health insurance data analysis on home nursing care in Hungary

Bevezetes: Az otthoni szakapolas 1996 vegen indult meg Magyarorszagon. Celkitűzes: A szerzők celja a magyar otthoni szakapolasi rendszer egeszsegbiztositasi es igenybeveteli mutatoinak bemutatasa. Modszer: Az elemzesben szereplő adatok az Orszagos Egeszsegbiztositasi Penztar finanszirozasi adatbazisabol (2001–2012) szarmaznak. Elemeztek a betegszamokat, a vizitszamokat, a szakapolasi es szakiranyu terapias tevekenysegek (gyogytorna, fizioterapia, logopedia) aranyat. Eredmenyek: Az ellatott betegek szama a 2001. evi 36 560-rol 2012-re 51 647-re nőtt, 41,3%-kal emelkedett. A teljesitett vizitek szama ugyanezen időszak alatt 841 715-ről 1 194 670-re, 41,9%-kal nőtt. A szakapolasi es szakiranyu terapias tevekenysegek, valamint az apolasi fokozatok vonatkozasaban jelentős megyek kozotti eltereseket figyeltek meg. A szakapolasra kifizetett osszegek aranya a legmagasabb volt Nograd (80,4%), Szabolcs-Szatmar-Bereg (79,7%) es Komarom-Esztergom (74,6%) megyekben, mig a legalacsonyabb Zala (53,0%) es Csongrad (52,7%...INTRODUCTION Home nursing care was introduced in Hungary in 1996. AIM The aim of this study was to analyse health insurance data and utilization indicators of the Hungarian home nursing care. METHOD Data derived from the database of the National Health Insurance Fund Administration (2001-2012). The number of patients and visits, and the ratio of special nursing and special therapy (physiotherapy, speech therapy) were analysed. RESULTS The number of patients increased by 41.3% from 36.560 (2001) to 51.647 (2012). The number of visits also increased by 41.9% from 841.715 (2011) to 1.194.670 (2012). Significant geographical inequalities were found in the ratio of special nursing and special therapy as well as nursing needs. The ratio of reimbursement for special nursing was the highest in county Nógrád (80.4%), Szabolcs-Szatmár-Bereg (79.7%) and Komárom-Esztergom (74.6%), while the lowest in county Zala (53.0%), Csongrád (52.7%) and Budapest (47.9%). CONCLUSIONS There are significant inequalities in the home nursing care in Hungary. In order to decrease these inequalities, specific guidelines should be developed for home nursing care.

Attendance of the fourth (2008-2009) screening round of the Hungarian organized, nationwide breast cancer screening program

INTRODUCTION Organised, nationwide screening for breast cancer with mammography in the age group between 45 and 65 years with 2 years screening interval started in Hungary in January 2002. AIM The aim of this study is to analyze the attendance rate of nationwide breast screening programme for the 2008-2009 years. METHOD The data derive from the database of the National Health Insurance Fund Administration. The ratio of women in the age group 45-65 years was calculated having either a screening mammography or a diagnostic mammography in the 4th screening round of the programme. RESULTS In the years 2000-2001, 7.6% of the women had an opportunistic screening mammography while in 2008-2009 31.2% of the target population had screening mammography within the organized programme. During the same periods 20.2% (2000-2001) and 20.4% (2008-2009) of women had a diagnostic mammography. Thus the total (screening and diagnostic) coverage of mammography increased from 26.6% (2000-2001) to 50.1% (2008-2009). The attendance rate failed to change between 2002 and 2009. CONCLUSIONS In order to decrease the mortality due to breast cancer, the attendance rate of mammography screening programme should be increased. Orv. Hetil., 154(50), 1975-1983.