Intergeneric hybrid origin of the invasive tetraploid Cirsium vulgare.

The invasive tetraploid Cirsium vulgare hybridizes with both Cirsium and Lophiolepis. Its conflicted position in molecular phylogenies, and its peculiar combination of morphological, anatomical, and genomic features that are alternatively shared with representatives of Cirsium or Lophiolepis, strongly suggest its intergeneric hybrid origin. Genetic relationships of C. vulgare (8 samples) with genus Lophiolepis (11 species) and other representatives of genus Cirsium (12 species) were evaluated using restriction site-associated DNA sequencing (RADseq) and examined using analytical and imaging approaches, such as NeighborNet, Heatmap, and STRUCTURE, to identify nuclear genomes admixture. Estimation of the intensity of spontaneous hybridization within and between Cirsium and Lophiolepis was based on herbarium revisions and published data for all reported hybrids pertinent to taxa currently included in Cirsium or Lophiolepis. The genome of any examined Cirsium species is more similar to C. vulgare than to any Lophiolepis species, and vice versa. The nuclear genome of the tetraploid C. vulgare is composed of two equivalent parts, each attributable either to Lophiolepis or to Cirsium; the organellar RADseq data clustered C. vulgare with the genus Cirsium. Spontaneous hybridization between Cirsium and Lophiolepis is significantly less intensive than within these genera. Our analyses provide compelling evidence that the invasive species C. vulgare has an allotetraploid intergeneric origin, with the maternal parent from Cirsium and the paternal from Lophiolepis. For the purpose of delimiting monophyletic genera, we propose keeping Lophiolepis separate from Cirsium and segregating C. vulgare into the hybridogenous genus Ascalea.

ASSESSMENT OF CORNEAL ENDOTHELIAL LAYER IN CONTACT LENS WEARERS WITH THE AID OF AN ENDOTHELIAL MICROSCOPE.

AIMS: The main aim of our study was to demonstrate the difference in endothelial cell density between a group of keratoconus contact lens users and non-contact lens users (without keratoconus). MATERIAL AND METHODS: In our study we had data from 96 subjects with an average age of 40.5 ±14.05 years. For the purposes of our study, we worked with each eye separately for each subject (n = 192). Keratoconus (research group) was diagnosed in 97 eyes. The mean age of the patients in the research group was 41.9 ±10.6 years. Keratoconus was not diagnosed in the remaining 95 eyes (control group). The mean age of the patients in the control group was 39.5 ±16.6 years. In the keratoconus group, the patients wore hard contact lenses (HCL) in a total of 43 eyes, hybrid contact lenses (HbCL) in 48 eyes and soft contact lenses (SCL) in 6 eyes. The average total period of contact lens use in the research group was 10.6 ±2.36 years. RESULTS: The average number of endothelial cells in the research group was 2607.11 ±298.45 cells per mm2. The average number of endothelial cells in the control group was 2831.94 ±523.51 cells per mm2. We tested these two variables using a T-test, which showed a statistically significant difference (p < 0.001). A statistically significant difference in the mean endothelial cell count related to patient age and use / non-use of contact lenses of two types was demonstrated only in the under-40 group in a comparison of the non-contact and keratoconus group with the keratoconus group using HCL (p = 0.02). A statistically significant difference in the length of contact lens wear was demonstrated between the keratoconus-free group with or without SCL and the keratoconus group when wearing HCL or HbCL for more than 20 years (p = 0.01 and p = 0.02). For HbCL users, this difference was demonstrated also after 15 years of wearing (p = 0.001). CONCLUSION: From our results we can conclude that there is a difference in endothelial cell density between patients with and without keratoconus.

SPECIFIC CORNEAL PARAMETERS AND VISUAL ACUITY CHANGES AFTER CORNEAL CROSSLINKING TREATMENT FOR PROGRESSIVE KERATOCONUS.

AIM: To evaluate the effect of crosslinking (CXL) therapy on the change in the quality of visual acuity and the change in the topographic properties of the cornea - curvature, pachymetry, and change of astigmatism, coma abberation and CLMIaa (Cone Localisation and Magnitude Index). METHODS: A retrospective analytical study included 29 eyes of 24 patients who had progressed in the last 12 months and were suitable candidates for CXL surgery. The monitored parameters were the steepest, flatest and mean anterior instantaneous curvature (AICS, AICF, AICM) and the steepest, flatest and mean posterior instantaneous curvature (PICS, PICF, PICM) of the cornea, corneal thickness in the centre of the cornea (PACHC) and in the thinnest point of the cornea (PACHT), corneal astigmatism (ASTIG). coma (COMA), Cone Localization and Magnitude Index (CLMIaa) and uncorrected distance visual acuity (UDVA) with corrected distance visual acuity (CDVA). Data were analysed before surgery and 12 months after surgery. The AIC, COMA, CLMIaa and ASTIG parameters were analysed by paired t test. As the parameters of UDVA, CDVA, PIC and PACH did not meet the conditions of normal distribution, the Wilcoxon test was used to investigate the change in these parameters after CXL. RESULTS: Twelve months after the procedure, we recorded an improvement in UDVA (p = 0.371) and CDVA (p = 0.825), an increase in PICS, PICF and PICM (p = 0.902; p = 0.87 and p = 0.555), a decrease in PACHCC (p = 0.294) and a decrease in CLMIaa (p = 0.113) that did not reach statistical significance. The decrease in PACHT (p = 0.027), decrease in COMA (p = 0.037) and decrease in anterior corneal curvature of AICS, AICF and AICM were statistically significant (p = 0.019; p = 0.010 and p = 0.005). The decrease in the value of astigmatism did not show statistical significance, as p = 0.297. CONCLUSION: CXL corneal therapy has been shown to be an effective method to stabilize the cornea in progressive keratoconus, and to improve the higher order of coma. This contributes to the possible improvement of UDVA and CDVA.

USAGE OF DIGITAL D CHART TEST AS A MODIFICATION OF AMSLER GRID IN OPHTHALMOLOGY AND OPTOMETRY.

AIMS: Metamorphopsia is important symptom of macular disease. The most common simple detection method of metamorphopsia is Amsler grid. Usually it is used monocularly with best correction for near. Patient should evaluate grid deformation a describe position of the deformity. This method is based on qualitative principle. For quantitative evaluation we can use Software D Chart (Thomson Software Solution). This instrument enables evaluate degree and position of the metamorphopsia in central visual field. Our goal was to establish M-score values in group of young healthy subjects without correction (M-score natural), with cylindrical spectacle lens (M-score SL) and in group of patients with age related degeneration (M-score ARMD). OBJECTS AND METHODS: We had 33 probands divided into 2 samples. The first sample contains 15 young probands with average age 23 years without any eye pathology. The second sample contains 18 patients with ARMD (7 with dry form and 11 with wet form). In our study we used software D Chart (Thomson Software Solution). This software was use in Acer PC with touchable screen. We note total M-score in right eye of all probands. Level for statistic evaluation was set on p = 0.05. RESULTS: Natural M-score values for young probands was: median 0, minimum 0, maximum 2.3. With cylindrical lens we got these values: median 25.2, minimum 3.6, maximum 41.6. In second sample with probands suffer from ARMD we got these values: median 0.8, minimum 0, maximum 29.4. Wilcoxon non-parametric test was used for statistical evaluation. We proved statistically significant difference between all variables. M-score natural vs. M-score SL showed p < 0.001, M-score natural vs. M-score ARMD showed p = 0.04 and M-score SL vs. M-score ARMD showed p < 0.001. CONCLUSION: Our study showed statistically significant differences between variable M-score natural, M-score SL and M-score ARMD. We found that printed Amsler grid as well as its digital modification D Chart are suitable for determining metamorphopsia in central visual field. The main advantage of D Chart is quantitative evaluation of the test with M-score and digital registration of retinal changes during patient´s follow up.

Sensitivity and specificity in methods for examination of the eye astigmatism.

PURPOSE: We usually use objective and subjective methods for examination of the eye astigmatism in optometry, respectively ophthalmology. Objective methods enable to measure sphere-cylindrical refraction of the eye. If we want to prescribe new glasses or contact lenses we usually use subjective methods. The aim of this study was to measure sensitivity and specificity of some subjective and objective methods for examination of the eye astigmatism. We supposed that automatic objective refraction will be the most exact method so we choose this method as the reference method. For comparison we chose subjective methods Jackson crossed cylinders (JCC), fogging method (FM) and objective method Spot Vision Screener (SVS, WelschAllyn). MATERIALS AND METHODS: We had in total 30 subjects with average age 23 years (SD 1 year) in our study. We made each measurement per eye separately and it was independent measurement so we could use measurement from each eye (n = 60). Each eye was firstly measured by subjective method FM, followed by JCC method and finally was use objective method Spot Vision Screener (SVS, WelschAllyn). Measurement with objective instrument TRK-1P (TOPCON) was use as reference measurement. The significance level was set at p = 0.05. RESULTS: In variable FM we measured sensitivity 76.2 % and specificity 66.7 %. Criterion for positive finding was -0.25 D. Result was statistically significant on level p < 0.001. In variable JCC we measured sensitivity 95.2 % and specificity 66.7 %. Criterion for positive finding was -0.25 D. Result was statistically significant on level p < 0.001. In variable SVS we measured sensitivity 47.6 % and specificity 94.4 %. Criterion for positive finding was -0.75 D. Result was statistically significant on level p < 0.001. Direct comparison of all methods showed statistically important difference between techniques JCC and FM (p = 0.0095). In other method we did not find statistically important difference (FM vs. SVS, p = 0.526 and JCC vs. SVS, p = 0.105). CONCLUSION: All subjective and objective techniques were statistically significant in detection of eye astigmatism. Comparison of ROC curves showed statistically significant difference between FM and JCC technique. The JCC method showed the highest sensitivity, whereas SVS highest specificity.

Digital Eye Strain in a Population of Young Subjects.

The main goal of our study was to prove short-term objective influence of near addition and relieving prism on accommodative and vergence facility in group of young healthy persons. Further, we wanted to prove objective and subjective difference between two types of soft contact lenses during working with digital device. We had in total 37 subjects in our study. The first group contained 37 subjects with average age 23 years without important eye pathology. The second group contained 8 subjects with the same average age without important eye pathology. In the first group of subjects, we measured binocular accommodative facility (BAF) and binocular vergence facility (BVF) with and without addition 1 D and with and without relieving prism 2 pD BO at working distance 45 cm. In the second group, we evaluated BAF, BVF and standardized questionnaire (Computer Vision Syndrome Questionnaire - CVS-Q, Sequí M. 2015) during usage of two types of soft contact lenses. The first type of contact lenses (type 1) contained relieving prism for PC working and type 2 was soft contact lens with aspheric design. In the first group of subjects, we measured these values: BAF without addition = 12.78 cpm, BAF with addition = 11.57 cpm, BVF without prism = 12.32 cpm, BVF with prism = 11.59 cpm. In the second group of subjects, we measured with contact lens type 1 these values: BAF = 12.13 cpm, BVF = 13.63 cpm and questionnaire score 9.43 points. With contact lens type 2 we measured BAF = 11.75 cpm, BVF = 11.63 cpm and questionnaire score 12.86 points. We proved statistically important different between two variable only in the first group between BAF with and without addition and BVF with and without relieving prism. In conclusion, we found statistically important decrease in variable BAF and BVF with usage of addition and relieving prism. We found that neither addition nor prism have positive influence on increase of BAF and BVF. Subjects in second group had higher BAF and BVF values with both types of contact lenses in comparison with natural values. We found that with contact lens type 1 (with addition) subjects had higher BAF and BVF values in comparison with contact lens type 2 (aspheric). Subjects with contact lens type 1 had also lower (better) questionnaire score, i.e. 9.43 versus 12.86 point. Key words: Digital eyestrain syndrome, accommodative and vergence facility, addition, relieving prism.

Sensitivity and specificity of spectral OCT in patients with early glaucoma.

This work deals with sensitivity and specificity in spectral OCT in detection of early glaucoma. Our goal was to evaluate data from RNFL analysis and compare them with resulted diagnosis of glaucoma neuropathy based on visual field changes. This retrospective study contains 31 subjects who undergone OCT examination in our department between years 2008 and 2017. Test statistic showed RNFL OCT sensitivity 63.64 % and specificity 90 %. Test ROC (receiver operating characteristic curve) showed AUC (area under curve) 0.734, which was statistically significant (p = 0.0097). We found that spectral OCT SLO with RNFL analysis is useful and effective instrument in analysis of glaucoma.

Colour vision in group of subjects with and without chromagen filter.

Our aim in this study was to prove influence of chromagen filter on color vision quality. Further we wanted to compare quality of color vision in groups of young healthy persons with persons with maculopathy. In our study we had in total 39 subjects. First group contained 13 subjects with average age 23 years without important eye pathology. In the second group we had 13 patients (average age 68 years) with maculopathy. Third group contained subjects with average age 64 years without important eye pathology. While examination we used sorting tests for color vision: Farnsworth-Munsell test and Lanthony test. Results were evaluated according the Vingryse and King-Smith technique. We found that average total error score (TES) in young healthy subjects with color chromagen filter doesnt differ from TES value gained from patients with disease of macula (p = 0.86). Further we found that chromagen filter changes color vision in group of young subjects on statistical significant level (p = 0.01). But in clinical view this is not important color vision defect (TES = 107.46 and CI = 1.42). Next study result showed statistically not important difference between the color vision in young healthy patient (average age 23 years) and older healthy patients with average age 64 years (p = 0.58). Finally we can conclude that green color chromagen filter doesnt have negative influence for dyslectic patients who will use this filter all day. Further we proved that color vision deterioration of patients with macular disease is not important for practical distinguishing of colors but is important clinically for diagnostic purposes. Last but not least we brought result which shows not important difference between young healthy subjects and older subjects (23 versus 64 years).Key words: Color vision, chromagen lens, total error score, maculopathy.

[Quantitative Color Vision Defect Evaluation - Lanthony Test and its Modified Interpretation]. / Kvantitativní vyhodnocení defektu barvocitu - Lanthonyho test a jeho modifikovaná interpretace.

UNLABELLED: Our aim was to develop quick and exact instrument for examination of color vision defects (CVD). We used Lanthony saturated and desaturated test. Data were evaluated according the Vingrys and King-Smith study. We had together 123 eyes of 86 patients. From all subjects we received these average values: AA 44.32 (min -87.13, max 80.64), TES 13.36 (min 8.84, max 30.30), SI 1.97 (min 1.22, max 5.69) and CI 1.66 (min 1.0, max 3.94). At the base of counting algorithm and average values form saturated and desaturated test we revealed 25 (29 %) patients with CVD. Twelve patients (14 %) classified as CVD+ had dichromacy and all had inborn CVD. Eight patients (9 %) from this group had deutranopia and four patient (5 %) protanopia. Anomaly trichromacy we revealed in thirteen patients (15 %). Eight (9 %) of these patients had inborn CVD. Six (7 %) of these patients had protanomalia, one (1 %) had deuteranomalia and one tritanomalia. We established and specified TES, CI and SI critical values, which was used to dividing patients into specific groups. KEY WORDS: Lanthony test, color vision defect, index of selectivity, index of confusion, total error score.

[Disorders of Simple Binocular Vision in Heterophoria and their Spectacle Correction]. / Poruchy jednoduchého binokulárního videní u heteroforií a jejich resení brýlovou korekcí.

This study deals with evaluation of binocular vision in group of young subjects without eye pathology. We examined at whole 68 subjects with average age 26 years, median 24 years. The sample was divided into two main groups. Group A contains subjects with far and near ortophoria (at whole 26 subjects = 38 %). Group B contains subjects with binocular vision disorder (at whole 42 subjects). One subject had strabismus. The most frequent non-strabismus disorder of binocular vision was convergence insufficiency (13 subjects = 19 %), simple esophoria (12 subjects = 17 %) and simple exophoria (8 subjects = 12 %). We decreased average distance heterophoria value (from 1.02 to 0.36 cm/m esophoria) and near heterophoria value (from 0.60 to 0.31 cm/m exophoria) after proper sphere-cylindrical correction. Result of our study shows that adequate and actual sphere-cylindrical correction can reduce disorder of simple binocular vision.Key words: simple binocular vision, heterophoria, spectacle lenses, convergence insufficiency.

Estimated Glomerular Filtration Rate in Oncology Patients before Cisplatin Chemotherapy.

BACKGROUND: The aim of the study is to compare measured glomerular filtration rate by technetium radiolabled diethylene tiamine pentaacetic acid (mGFR DTPA) to estimated GFR (eGFR). Glomerular filtration rate (GFR) is estimated from serum creatinine (eGFRcreatinine), serum cystatin C (eGFRcystatin C) and by combined equation (eGFRcreatinine+cystatin C). This study focuses on oncology patients considered for treatment with cisdiamminedichloroplatinum (cisplatin). We evaluated the impact of different GFR methods on the reduction of cisplatin dose. PATIENTS AND METHODS: The study population consisted of 112 consecutive oncology patients from oncology center treated in the town of Zlin in the Czech Republic, who were considered for cisplatin treatment. mGFR DTPA was performed by dynamic renal 99mTc scintigraphy method using diethyltriaminepentaacetic acid. Creatinine and cystatin C were determined by newly standardized tests. Estimation of GFR was calculated using The Chronic Kidney Disease Epidemiology (CKD EPI) equations which were established in 2009 and 2012. RESULTS: The median (interquartile range) of mGFR DTPA was 1.335 ml/s/1.73 m2 (1.070-1.725). The median of eGFRcystatin C 1.195 ml/s/1.73 m2 (0.885-1.625) was lower than mGFR DTPA (p<0.05). The median of eGFRcreatinine 1.460 ml/s/1.73 m2 (1.210-1.660) was higher than mGFR DTPA (p<0.05). Correlation analysis and Bland Altman plots show high individual differences between mGFR DTPA and all eGFRs. CONCLUSIONS: Oncology patients are a very special group of patients who dif-fer from general population. There are significant individual differences between mGFR DTPA and all eGFRs, impacting detection rate of CKD and potential drug dosage adjustment.

[Stereo Vision Deterioration by Artificially Induced Aniseikonia]. / Zhorsení prostorového videní umele navozenou aniseikonií.

UNLABELLED: Main purpose of this study was to evaluate effect of aniseikonia on the stereo vision. We had together 90 subjects without eye pathology with or without habitual correction. Five of them were excluded due to important anisometropia or bad visual acuity (V < 0.5 on worse eye). All 85 subjects every in 4 cases (without size lens, with size lens on OD 1, 3 and 5 %) undergone measuring of their stereoscopy parallax. This was evaluated by Random dot stereo test. The level for stereoscopy vision was set bellow 60 arc seconds. This criterion was not achieved naturally by 6 subjects, so final number of all cases was 316 (100 %). As a whole 48 subjects (15.2 %) fail after using the test with size lens on OD 1, 3 or 5 %. All 268 cases (84.8 %) had not impaired stereoscopy parallax with size lens over chosen critical level. KEY WORDS: size lens, anisometropia, aniseikonia, heterophoria, stereoscopy vision.

[First Results and Experience with the iTrace Aberometer]. / První výsledky a zkusenosti s aberometrem iTrace.

PURPOSE: To evaluate obtained data by using a relatively novel devices and their results which are important eg. in refractive and cataract surgery. MATERIAL AND METHODS: The study included 66 eyes (n = 66). Subjects were represented by 32 women and 1 man whose age was 22.5 years ± 1.2 years (min. 21, max. 26 years) without any signs of potential eye disease. Duration of the study was 3 months. Results were compared with the measurements using the auto-refract-keratro-tono-pachymeter (TRK 1P, Topcon, Japan), Keratograph 5M (Oculus, Germany) and aberrometer iTRACE (Hoya, Japan). RESULTS: After 3 months were statistically compared keratometry values of ​​ corneal anterior surface detected by all devices. They provided to be comparable. Furthermore the values ​​of objective refraction and pachymetry were detected. CONCLUSION: Results of this study show a statistically significant correlation values ​​of objective refraction using devices TRK and iTRACE (r = 0.66 at p = 0.05) and showed a significant relationship between the keratometric data for all the devices. All used methods and devices are possible to reliably and use for valid evaluation parameters of the eye. KEY WORDS: aberrometry, low and high orders abberations, keratometry, topography, pachymetry.

Egg yolk phospholipids enriched with 1-O-octadecyl-2-oleoyl-sn-glycero-3-phospho-(N-palmitoyl) ethanolamine inhibit development of experimentally induced tumours.

Dietary phospholipids (PLs) and their derivatives have proved active in suppression of various health problems and conditions including cancer. In this work we compared the effect of dietary phospholipids from hen egg yolk enriched with N-acyl ether-phosphatidyl ethanolamine (NAEPE) termed bioactive phospholipids (BAP+ preparation) with PLs lacking NAEPE (BAP- preparation) on the growth of transformed cells in vitro and on the promotion and progression of experimental tumours in vivo. For the in vivo experiments we used the chicken model in which liver, lung, and kidney tumours arose via natural selection from single cells initiated by experimentally introduced somatic mutations caused by insertional mutagenesis. Mutagenized animals were fed BAP+ or BAP- diet in various regimens. We observed that BAP+ at low concentrations killed cells of various tumour cell lines in culture but did not compromise viability of non-transformed cells. Oral administration of the BAP+ preparation efficiently reduced progression of all tumour types. However, it did not significantly reduce the number of already initiated tumours and their growth when BAP+ was discontinued. Our data suggest that NAEPE combined with hen egg PLs significantly interferes with tumour progression, possibly through the inhibition of tumour cell viability.

[Simple binocular vision evaluation on healthy adult subjects with synoptophore]. / Evaluace parametru jednoduchého binokulárního videní na synoptoforu u zdravé dospelé populace.

PURPOSE: The main goal of our study was to determine the database of parameters of simple binocular vision (SBV) in healthy adult population. Next goal was to verify current data of particular parameters of SBV. Recent public data (e.g. Divisova, Hromadkova) were determined without proper specification of examination (size of fusion object), or are too diffused (e.g. from 15 to 25 prismatic diopters = pD). At last we want to prove, if there are some other factors, which could influence parameters of SBV. METHODS: We had 74 subjects (64 women, 10 men) without significant eye pathology with average age of 24.82 years (max. 28 years, min. 22 years, SD 2.5 years). Particular parameters of SBV were examined without corrective lenses by emetropes, but with habitual correction by ametropes all on Sbisa synoptophore. We measured minimally 16 parameters of SBV, which were note in centimeters (pupillary distance) and in prismatic dioptres (deviation, fusion range and so on). These data were than statistically processed with program MS Excel and with Statitstika version 10. Level of statistical significance was set on p = 0.05. RESULTS: We got, thanks our study, these significant average results: Subjective deviation for far of all 74 subjects was count to 2.78 ± 3.65 pD, of women was 2.90 ± 3.69 and only for men was 2.00 ± 3.49. This result probably shows inadequate elimination of proximal convergence with the instrument. According our measurement of positive fusion range of all subjects is 25.10 ± 12.77 pD and negative fusion range -6.45 ± 4.18 dD, accommodation convergence to accommodation ration (AC/A) is 3.41 ± 1.47 pD a subjective deviation by accommodation on 33 cm (with minus 3 D) is 13.02 ± 5.23 pD. Further we proved statistical significant correlation between these parameters of SBV: Age and AC/A, SU-3 and AC/A, SU0 and SU-3, SU-3 and FS0 and FS-3. We didnt find any statistical significant differences, when we compared SBV parameters between emetropes, hypermetropes and myopes. The same results we got (except of pupillary distance), when we compared data divided according to gender. CONCLUSION: In our study we set the normative average data of SBV parameters, which were measured on healthy adult emetropes and ametropes with habitual correction. We also find how parameters influence each other. All SBV parameters differ on statistical significant level, when we compared them with respect to refractive state and gender. Knowledge of basic SBV parameters is important not only for ophthalmologist but also for optometrists. They can influence these in that way, which can bring comfortable SBV. The most frequent optometric methods are proper sphere-cylindrical correction, prismatic correction and visual training. accommodation ration, pupillary distance.

Dynamics and morphology of focal adhesions in complex 3D environment.

Focal adhesions are specific types of cellular adhesion structures through which both mechanical force and regulatory signals are transmitted. Recently, the existence of focal adhesions in 3D environment has been questioned. Using a unique life-like model of dermis-based matrix we analysed the presence of focal adhesions in a complex 3D environment. Although the dermis-based matrix constitutes a 3D environment, the interface of cell-to-matrix contacts on thick bundled fibres within this matrix resembles 2D conditions. We call this a quasi-2D situation. We suggest that the quasi-2D interface of cell-to-matrix contacts constituted in the dermis-based matrix is much closer to in tissue conditions than the meshed structure of mostly uniform thin fibres in the gel-based matrices. In agreement with our assumption, we found that the cell adhesion structures are formed by cells that invade the dermis-based matrix and that these structures are of similar size as focal adhesions formed on fibronectin-coated coverslips (2D). In both 2D situation and the dermis-based matrix, we observed comparable vinculin dynamics in focal adhesions and comparable enlargement of the focal adhesions in response to a MEK inhibitor. We conclude that focal adhesions that are formed in the 3D environment are similar in size and dynamics as those seen in the 2D setting.

Local treatment of hand-foot syndrome with uridine/thymidine: in vitro appraisal on a human keratinocyte cell line HaCaT.

5-fluorouracil (5-FU) is one of the most commonly used antineoplastic drugs in the anticancer therapy. The hand-foot (HF) syndrome (palmar-plantar erythrodysesthesia) is an adverse effect frequently related to long-term i.v. administration of 5-FU or its orally applicable prodrug capecitabine. Its severity can even lead to interruption of the otherwise effective anticancer therapy. Tentative practice in some clinics has shown that topical application of 10% uridine ointment is beneficial for calming down the HF syndrome. This study is focused on verifying the alleged protective activity of uridine in the in vitro model of cultured human keratinocyte cell line HaCaT. We also tested the protective effects of thymidine alone or uridine-thymidine combination. The cellular viability time progression was measured in order to evaluate the effect of protective agents by three different types of cytopathogenicity tests-NTCA test (non-destructive test of cellular activity), modified MTT test and RTCA (real-time cell analyser, Roche). All three methods proved the ability of uridine and uridine-thymidine combination to protect keratinocytes against 5-FU damage in vitro. While thymidine alone did not show any remarkable effect, the thymidine-uridine combination demonstrated enhanced protective activity compared to uridine alone. Our findings provided the supporting rationale for using uridine or uridine-thymidine ointments in the HF syndrome local therapy.

[Repeatability and reliability of the visual acuity examination on logMAR ETDRS and Snellen chart]. / Opakovatelnost a spolehlivost vysetrení zrakové ostrosti na optotypových tabulích logMAR ETDRS a Snellen.

PURPOSE: The main goal of our study was to prove the statistical significant correlation between repeated measurements and test-retest variability TRV expressed with confidence interval CI according to Bland-Altman's method on 95% level of statistical confidence. The methods are threshold interpolation logMAR method on ETDRS chart, whole-line method on Snellen chart and modified threshold interpolation method on Snellen chart with Sloan letters. METHODS: We had 468 measurements measured with threshold interpolation log MAR method on ETDRS chart, whole-line method on Snellen chart and modified threshold interpolation method on Snellen chart with Sloan letters. The average value of the first sequence of measurements measured with whole-line method on Snellen chart was -0.043 logMAR (min. 1, max. -0.30. SD 0.25) and of the second sequence of measurements was -0.045 logMAR (min. 1, max. -0.30. SD 0.23). The average value of the first sequence of measurements measured with interpolation method on Snellen chart was -0.018 logMAR (min. 0.98, max. -0.30. SD 0.29) and of the second sequence of measurements was -0.024 logMAR (min. 1, max. -0.80. SD 0.29). The average value of the first sequence of measurements measured with interpolation method on ETDRS chart was -0.0612 logMAR (min. 0.72, max. -0.30. SD 0.21) and of the second sequence of measurements was -0.0610 log MAR (min. 0.8, max. -0.28, SD 0.21). RESULTS: We have proved that all methods do not have statistical significant difference between repeated measurements (Wilcoxon paired test, whole-line method on Snellen chart p = 0.74, interpolation method on Snellen chart p = 0.33 and interpolation method on ETDRS p = 0.95) and they also have statistical significant correlations (Spearman correlation coefficient, whole-line method on Snellen chart r = 0.91, p < 0.0001, interpolation method on Snellen chart r = 0.89, p < 0.0001 and interpolation method on ETDRS chart r = 0.89, p < 0.0001). TRV expressed with CI on 95% statistical significance level according to method of Bland-Altman was with whole-line method on Snellen chart +/- 0.11 (i.e., 5 letters), with interpolation method on Snellen chart +/- 0.20 (i.e., 10 letters) and with interpolation method on ETDRS +/- 0.08 (i.e., 4 letters). CONCLUSION: In our study we proved that there is no statistical significant difference between repeated measurements with all three methods. After that we proved statistical significant correlations between repeated measurements with all three methods. The biggest clinical importance has determination and confirmation of the CI value of TRV. With interpolation method on ETDRS it was CI +/- 0.08 (i.e., +/- 4 letters), with whole-line method on Snellen it was CI +/- 0.11 (i.e., +/- 5 letters) and with interpolation method on Snellen it was CI +/- 0.20 (i.e., +/- 10 letters). Our recommendation based on results of our study is that is not clinically appropriate to convert measured values from Snellen optotype to logMAR and perform interpolation method on Snellen chart.

Skeletal muscle damage and impaired regeneration due to LPL-mediated lipotoxicity.

According to the concept of lipotoxicity, ectopic accumulation of lipids in non-adipose tissue induces pathological changes. The most prominent effects are seen in fatty liver disease, lipid cardiomyopathy, non-insulin-dependent diabetes mellitus, insulin resistance and skeletal muscle myopathy. We used the MCK(m)-hLPL mouse distinguished by skeletal and cardiac muscle-specific human lipoprotein lipase (hLPL) overexpression to investigate effects of lipid overload in skeletal muscle. We were intrigued to find that ectopic lipid accumulation induced proteasomal activity, apoptosis and skeletal muscle damage. In line with these findings we observed reduced Musculus gastrocnemius and Musculus quadriceps mass in transgenic animals, accompanied by severely impaired physical endurance. We suggest that muscle loss was aggravated by impaired muscle regeneration as evidenced by reduced cross-sectional area of regenerating myofibers after cardiotoxin-induced injury in MCK(m)-hLPL mice. Similarly, an almost complete loss of myogenic potential was observed in C2C12 murine myoblasts upon overexpression of LPL. Our findings directly link lipid overload to muscle damage, impaired regeneration and loss of performance. These findings support the concept of lipotoxicity and are a further step to explain pathological effects seen in muscle of obese patients, patients with the metabolic syndrome and patients with cancer-associated cachexia.

[Comparison of the threshold interpolation and whole-line method on logMAR chart and Snellen chart for visual acuity testing]. / Porovádní prahové interpolacní a celorádkové metody pri testování zrakové ostrosti na optotypových tabulich logMUR ETDRS a Snellen.

The main goal of our study was to prove the statistical significant difference between the threshold interpolation logMAR method on ETDRS chart and the whole-line method on Snellen chart with Sloan letters. We had 108 measurements with the threshold interpolation method and the whole-line method on ETDRS chart and the whole-line method on Snellen chart. The average value measured with the threshold method in ETDRS was 1,132 (min. 0,660, max. 1,580), with the whole-line method on ETDRS it was 1,134 (min. 0,630, max. 1,580) and with the whole-line method on Snellen chart it was 1,183 (min. 0,630, max. 1,600). We have proved statistical significant difference between the threshold interpolation method made on ETDRS chart and the whole-line method made on Snellen chart (p < 0.001). The values measured with the whole-line method on Snellen chart were overvalued. The exact and reliable measuring of visual acuity is an important component of further examinations (e.g. contrast sensitivity, perimetry, tonometry), which enable us to make a correct diagnosis of pathological changes on human eye structures.