LUCENTIS IN THE TREATMENT OF DIABETIC MACULAR EDEMA, TWO-YEAR RESULTS.

AIM: To evaluate, on the basis of two-year observations, the effectiveness of intravitreal treatment with Ranibizumab in patients with diabetic macular edema (DME) unresponsive to the previous laser treatment. Cohort and Methods: A retrospective study evaluates 29 eyes of 29 patients with diffuse DME unresponsive to their previous laser treatment. The group of the patients consisted of 16 males (55.1%) and 13 females (44.8%); their mean age was 71.3. The mean duration of diabetes mellitus was 13 years (3-20). 19 patients (65.5%) were treated with insulin, 10 patients (34.4%) were treated with peroral antidiabetics (PAD); the mean HbA1c value was 52 mmol/l. The treatment was started with 3 initial doses of intravitreal injections of Ranibizumab 0.5 mg. There was a one- -month interval between the applications. Subsequent evaluations and administrations of the following injections were made in the pro re nata (PRN) mode; the check-ups were carried out every month during the first year and on average every 3 months in the second year. The monitored parameters: the best corrected visual acuity (BCVA) measured on ETRDS (Early Treatment Diabetic Retinopathy Study) optotypes, the central retinal thickness (CRT). These parameters were monitored prior to the treatment and then in the 3rd, 6th, 9th, 12th, 18th and 24th months. RESULTS: A statistically significant improvement in the mean value of BCVA was detected. From the initial 65.4 ±10.61 letters it improved by 11.2 letters (p.

The use of micropulse laser in patients with diabetic macular edema at the Department of Ophthalmology, Faculty Hospital Hradec Králové.

INTRODUCTION: Diabetic macular edema (DME) is the most frequent cause of decreased vision in patiens with diabetes type 2. DME is caused by increased permeability of macular capillaries. The aim of our study was to retrospectively evaluate the results of micropulse laser treatment, 577 nm wavelength, in pacients with DME with follow-up three months and one year. METHODS: The retrospective trial with one year follow-up includes 63 eyes of 37 patients with diabetic macular edema treated from September 2015 to January 2017.  Most patients had diabetes type 2 (34 patients), 3 patients had diabetes type 1. Diffuse DME was diagnosed in 54 eyes, focal DME in 9 eyes. Most of the patients (42 eyes) suffered from non proliferative diabetic retinopathy, 21 eyes showed signs of proliferative retinopathy. DME lasted on average for 29 months before initiating with micropulse laser (median 21 months). On average 1,56 laser visits were needed for the treatment of 1 eye, usually in 3 months interval. Photocoagulation of macula was performed in all patients by micropulse laser, 577 nm wavelenght/IQ 577TM IRIDEX). We used 5 % duty cycle. The average glycated hemoglobin in the group was 66,8 mmol/mol, maximal 100 mmol/mol. Estimated data were statistically evaluated by Friedman and Dunn´s test. RESULTS: At the end of 1 year period we found out improvement in BCVA (increase of at least 5 letters of ETDRS charts) in 20 eyes, 25 eyes showed stabilisation of BCVA (alltogether 71 % of the group), in 18 eyes we found out decrease of BCVA of more than 5 letters of ETDRS charts. On average we estimated decrease of visual acuity from 62 to 61,1 letters (p > 0,05). After one year we estimated 63 µm CRT decrease on average, from 442 µm to 379 µm (p= 0,0124). CONCLUSION: In our group of DME patiens treated by micropulse laser we have estimated BCVA stabilisation and signifiant improvement of macular edema in ¾ eyes, confirmed by OCT. We have estimated clinically significant decrease of macular edema in the whole group with one-year follow-up.

Cells transformed by murine herpesvirus 68 (MHV-68) release compounds with transforming and transformed phenotype suppressing activity resembling growth factors.

In this study, we investigated the medium of three cell lines transformed with murine herpesvirus 68 (MHV-68) in vitro and in vivo, 68/HDF, 68/NIH3T3, and S11E, for the presence of compounds resembling growth factors of some herpesviruses which have displayed transforming and transformed phenotype suppressing activity in normal and tumor cells. When any of spent medium was added to cell culture we observed the onset of transformed phenotype in baby hamster kidney cells (BHK-21) cells and transformed phenotype suppressing activity in tumor human epithelial cells (HeLa). In media tested, we have identified the presence of putative growth factor related to MHV-68 (MHGF-68). Its bivalent properties have been blocked entirely by antisera against MHV-68 and two monoclonal antibodies against glycoprotein B (gB) of MHV-68 suggesting viral origin of MHGF-68. The results of initial efforts to separate MHGF-68 on FPLC Sephadex G15 column in the absence of salts revealed the loss of its transforming activity but transformed phenotype suppressing activity retained. On the other hand, the use of methanol-water mobile phase on RP-HPLC C18 column allowed separation of MHGF-68 to two compounds. Both separated fractions, had only the transforming activity to normal cells. Further experiments exploring the nature and the structure of hitherto unknown MHGF-68 are now in the progress to characterize its molecular and biological properties.

[Wet form age-related macular degeneration two years treatment results using anti VEGF drugs]. / Dvouleté výsledky lécby vlhké formy vekem podmínené makulární degenerace s pouzitím léku proti vaskulárnímu endotelovému rustovému faktoru.

AIM: The aim of the study was to establish the efficacy of anti VEGF (Vascular Endothelial Growing Factor) drugs in the treatment of wet form ARMD (Age-Related Macular Degeneration) in everyday clinical practice in the Department of Ophthalmology, Faculty Hospital, Hradec Králové, Czech Republic, E.U., in patients registered in the Czech national registry AMADEUS. MATERIAL AND METHODS: Retrospective study with 24 months follow-up period. In the group were evaluated 143 eyes of 140 patients, out of them were 77 women (65.8 %), of average age 73.09 (71.69 - 74.48) years, and 40 men (34.2 %) of average age 74 (58 - 85) years. All of the patients were completely examined before the beginning of the treatment; during the treatment were, except the standardized eye examination, in patients treated with ranibizumab the color fundus photography and Optical Coherence Tomography (OCT) with measuring of the central retinal thickness performed every three months at least. The patients treated by pegaptanib were examined every six weeks before the drug application. The fluorescence angiography (FA) was performed at the beginning of the treatment to establish the type and extension of the choroidal neovascularization and during the treatment in case of necessity to establish the activity of the choroidal neovascular membrane (CNV). The treatment by ranibizumab was in the regimen PRN (pro re nata), and pegaptanib was applied every six months during the first year with the follow-up evaluation of the findings. The treatment evaluations were performed at 12 and 24 months. RESULTS: During the two years follow - up period, the authors noticed in patients treated with ranibizumab loss of 5.12 letters of ETDRS optotypes in case of mostly classical CNV, in occult CNV loss of 5.45 letters, and in minimally classical CNV loss of 2.83 letters. In three evaluated eyes with classical CNV in patients treated with pegaptanib we noticed after 2 years loss of 6.67 letters, in eleven eyes with occult CNV we established loss of 9.91 letters, and in two eyes with minimally classical CNV the average best-corrected visual acuity (BCVA) remained unchanged. The pegaptanib treatment results may be influenced by small number of evaluated patients. The visual acuity changes during the two years treatment were not statistically significant. We noticed the decrease of average CRT (central retinal thickness) in all types of CNV treated both with ranibizumab and pegaptanib after the two years follow up. To reach these results, an average of 5.51 applications of ranibizumab and 9 applications of pegaptanib during the two years were used. CONCLUSION: In the followed-up group we found, comparing to the natural course of neovascular form of ARMD, retarding of the BCVA decrease during the two years treatment with VEGF inhibitors in everyday clinical practice. Better results were achieved with ranibizumab treatment, however the differences were not statistically significant. Key words: age related macular degeneration, AMADEUS Czech national registry, ranibizumab, pegaptanib.

[The coincidence of the junction layer of inner and outer photoreceptors segments (IS/OS) defects localization on SD OCT and functional defects in case of Stargardt disease--a case report]. / Koincidence lokalizace defektu vrstvy junkce IS/OS fotoreceptoru na SD OCT s funkcními poruchami v prípade Stargardtovy choroby.

Case report of the different visual functions of the right and left eye of 60-years old man with Stargardt disease is presented. The difference in best corrected visual acuity between both eyes is accompanied with corresponding various alteration of the central retinal activity using multifocal electroretinography (mfERG) and asymmetrical defects of the IS/OS photoreceptor junction using HD-OCT.

Capillary zone electrophoresis of orotic acid in urine with on-line isotachophoresis sample pretreatment and diode array detection.

Potentialities of capillary zone electrophoresis with on-line isotachophoresis sample pretreatment and diode array detection (ITP-CZE-DAD) to the separation, detection and identification of trace analytes present in biological matrices were investigated. Urine represented a multicomponent, variable and high ionic strength matrix while orotic acid was chosen as a model analyte of a practical clinical relevance in this investigation. Using the ITP-CZE combination in the column-coupling configuration of the separation system ITP provided an enhanced sample load capacity to the separation system (a 30 microl sample injection volume), concentrated the analyte and served as an on-line sample clean up technique. On the other hand, CZE performed a final separation of the analyte from matrix constituents present in the ITP pretreated sample and provided favorable conditions for its detection and identification by DAD. Using current correction and smoothing procedures analytically relevant DAD spectra of orotic acid could be obtained also in instances when this was injected in a model sample at a 2 x 10(-7) mol/l concentration (an estimated limit of determination of orotic acid at a 218 nm detection wavelength). ITP-CZE separations of urine samples (based on differences in acid-base properties and host-guest complexations of the analyte and matrix anionic constituents) led to significant sample clean ups. Consequently, DAD spectra of orotic acid matching its reference spectrum, could be acquired also in instances when the acid was present in urine matrices (loaded in 30 microl injection volumes of 20-fold diluted urine samples) at 4-6 x 10(-7) mol/l concentrations. Here, residual trace matrix interferents prevented a closer approach to the above value attainable for model samples. Although this work was focused only on one analyte and urine matrix it implies very promising potentialities of the ITP-CZE-DAD combination in the identification and quantitation of trace analytes present in biological matrices, in general.

Capillary electrophoresis of inorganic anions.

This review deals with the separation mechanisms applied to the separation of inorganic anions by capillary electrophoresis (CE) techniques. It covers various CE techniques that are suitable for the separation and/or determination of inorganic anions in various matrices, including capillary zone electrophoresis, micellar electrokinetic chromatography, electrochromatography and capillary isotachophoresis. Detection and sample preparation techniques used in CE separations are also reviewed. An extensive part of this review deals with applications of CE techniques in various fields (environmental, food and plant materials, biological and biomedical, technical materials and industrial processes). Attention is paid to speciations of anions of arsenic, selenium, chromium, phosphorus, sulfur and halogen elements by CE.

Endothelial cell adaptation to chronic thrombosis.

BACKGROUND: The autogenous vein graft has proven to be the most durable conduit in lower extremity vascular bypass grafts. Failures due to thrombosis, intimal hyperplasia, and progression of atherosclerotic disease commonly plague the vascular surgeon. Part of the ability of vein grafts to provide a nonthrombogenic surface relies on the capability of the endothelial cell to produce prostacyclin, a potent vasodilator and inhibitor of platelet aggregation. Once a graft fails and thromboses, little is known as to the effects of the thrombus on the function and morphology of endothelial cells. Earlier studies by this laboratory demonstrated the ability of arterialized canine vein grafts to recover function after 5 days of exposure to thrombus. This investigation sought to explore the limits of endothelial cell viability and recovery to extended periods of thrombosis. METHODS: Using a canine model of arterialized vein grafts, prostacyclin production (measured as 6-keto-PGF1a) was assessed in an ex vivo perfusion system from grafts exposed to thrombus for 10 days (group I) and 20 days (group II). Both groups underwent thrombectomy and a recovery period of 30 days. The grafts were perfused with Hanks' balanced salt solution and samples were obtained at 5 and 30 minutes to determine prostacyclin levels. Arachidonic acid was then added to a new perfusate of Hanks' solution and samples were again obtained at 5 and 30 minutes. Results were expressed as PGF/graft area (cm2/min). Representative samples of each graft underwent scanning electron microscopy. RESULTS: Without arachidonic acid, prostacyclin production of group II (20 day) grafts was greater than group I (10 day) grafts at 5 minutes of perfusion (4.31 versus 2.42, P = 0.08) and at 30 minutes (1.86 versus 0.95, P = 0.02). In response to the addition of arachidonic acid both groups increased prostacyclin production (group I, P = 0.004; group II, P = 0.12). A comparison was made between prostacyclin production at baseline and after addition of arachidonic acid. Group I grafts demonstrated a greater percent increase in prostacyclin production compared to group II (385% versus 229%, P = 0.01). Scanning electron microscopy showed no differences in endothelial coverage between the study groups. CONCLUSIONS: These results demonstrate that although endothelial cells are able to recover a basal level of prostacyclin production, the response to substrate stimulation diminishes with increased exposure time to thrombus. This diminished response may be important in understanding the ability of vein grafts to survive after a period of thrombosis.

The effect of thrombus on the vascular endothelium of arterialized vein grafts.

BACKGROUND: It is known that vein grafts can be salvaged by clot removal, but patency rates are diminished. This study was designed to determine the effects of thrombus on vascular endothelium and the ability of the endothelium to recover normal function. METHODS: Thirty external jugular vein grafts were placed as bilateral femoral artery interposition grafts in 15 mongrel dogs and allowed to arterialize for a period of at least 12 weeks. Six control grafts were not exposed to thrombus (C-NT). Six other control grafts were exposed to thrombus for 7 days and removed, ie, allowed no in vivo recovery (C-T). The remaining 18 grafts in 9 canines were exposed to autologous thrombus for 5 days and then flow was restored. The right femoral graft was removed 7 days after thrombectomy and the left removed 30 days after thrombectomy. At the time of removal, the grafts were perfused with a balanced salt solution alone and then with arachidonic acid added to the same volume of the salt solution. Perfusates were collected at 5, 15, and 30 minutes. These perfusates were assayed for the presence of 6-keto-prosglandin F1 alpha (6-keto-PGF1(1 alpha)), a metabolite of prostacyclin (PGI2). Over the 30-day recovery period, the amounts of 6-keto-PGF1(1 alpha) produced with and without arachidonic acid added were compared to assess endothelial response. Electron micrographs of the endothelium of all vein grafts were compared to the assay findings. RESULTS: When arachidonic acid was added to the perfusion system, there was a several fold increase in the production of 6-keto-PGF1(1 alpha) over baseline in all grafts allowed recovery. Grafts (C-T) that were allowed no in vivo recovery had no response to arachidonic acid. Ratios of 6-keto-PGF1(1 alpha) production with arachidonic acid stimulation to 6-keto-PGF1(1 alpha) production without stimulation were calculated to compare endothelial function. The electron micrographs showed the vascular endothelium to be severely injured after contact with thrombus, but recovered by 7 days. CONCLUSIONS: This study suggests that the endothelium of canine vein grafts is injured by contact with thrombus for 5 days but can recover structure and function. This recovery is detectable at 7 days post-thrombectomy.

A comparison of some metabolitesin skeletal muscle of different halothane sensitivity withmeat quality traits of pigs using a capillary isotachophoresis technique.

Isotachophoretic (ITP) measurements were made on biopsy samples from live muscle M. longissimus lumborum and carcasses of pigs. Two experiments were performed. In Experiment 1, the methodological aspects of ITP measurements from skeletal muscle, were studied. In Experiment 2, ITP measurements on muscle and carcass samples were made. The muscle and carcass samples were obtained by shot or spring-loaded biopsy from the Longissimus lumborum muscle of 30 Belgium Landrace × Duroc pigs, of which 10 were halothane-positive. The pigs were slaughtered by electro-stunning and manguination at approximately 105 kg body l. w. The potential meat quality in live pigs and after slaughter using small biopsy samples of M. longissimus lumborum was also determined. The experimental results show that ITP (mainly inosinemonophosphate and lactate) and meat quality data (water-holding capacity test, pH and R value) can differentiate halo thane-positive from halothane-negative pigs. Out results confirmed previous results which showed that the water-holding capacity test defined as fluid volume', pH and R value measurements on biopsy samples can also predict the potential meat quality in live pigs.

Photometric detection at 405 nm in trace analysis by capillary isotachophoresis.

The possibilities of photometric detection at 405 nm were investigated for trace isotachophoretic analysis of various ionic constituents. Increased selectivity of the detection enabled, for example, the detection and determination of nitrophenols at 10(-8) M in a 25-microliter sample. 5-Nitrofurylacrylic acid, a wine stabilizer, could be detected with confidence at ca. 0.1 ppm in wine without any sample pretreatment. Approximately 250-fmol amounts of amino acids, labelled with 2,4-dinitrophenyl, were detectable with the technique employed. This is almost three decades lower in comparison to what is achievable with an high resolution universal detector. The adsorption of the analytes in the sample handling devices as well as in the separation compartment were problems in the analysis at this concentration level. The detection limit given above was achieved when these phenomena were suppressed by the addition of naphthalene-1,3,6-trisulphonate or pyrophosphate to the sample solution.

A fiber apparatus in the nucleus of the yeast cell.

The structure and mode of division of the nucleus of budding yeast cells have been studied by phase-contrast microscopy during life and by ordinary microscopy after Helly fixation. The components of the nucleus were differentially stained by the Feulgen procedure, with Giemsa solution after hydrolysis, and with iron alum haematoxylin. New information was obtained in cells fixed in Helly's by directly staining them with 0.005% acid fuchsin in 1% acetic acid in water. Electron micrographs have been made of sections of cells that were first fixed with 3% glutaraldehyde, then divested of their walls with snail juice, and postfixed with osmium tetroxide. Light and electron microscopy have given concordant information about the organization of the yeast nucleus. A peripheral segment of the nucleus is occupied by relatively dense matter (the "peripheral cluster" of Mundkur) which is Feulgen negative. The greater part of the nucleus is filled with fine-grained Feulgen-positive matter of low density in which chromosomes could not be identified. Chromosomes become visible in this region under the light microscope at meiosis. In the chromatin lies a short fiber with strong affinity for acid fuchsin. The nucleus divides by elongation and constriction, and during this process the fiber becomes long and thin. Electron microscopy has resolved it into a bundle of dark-edged 150 to 180 A filaments which extends between "centriolar plaques" that are attached to the nuclear envelope.