Use of ex-press® implant in glaucoma surgery - retrospective study.

PURPOSE: The objective of the study is to evaluate of primary open-angle glaucoma (POAG) with the use of EX-PRESS drainage implant. Evaluated was the decrease of intraocular pressure (IOP), visus stabilization, perimeter Humphrey - related finding (T 30-2), Heidelberg Retina Tomograph (HRT) and possibility of reducing the local drug therapy. PATIENTS AND METHODS: Retrospective data analysis was performed in 40 eyes with POAG in 28 subjects (14 female- and 14 male patients) average-aged 69.5 years. In all 40 eyes, surgery was performed by one surgeon within the years 2011-2017. Indications for EX-PRESS implantation in our study were the POAG with decompensated IOP, decompensated chronic secondary glaucoma or failure previously anti-glaucoma surgical operations. Within the preoperative period, in all cases the progression was observed on the perimeter T 30-2 or on the HRT. Before and after surgery, all 40 eyes were evaluated for the following factors: the IOP, visus, pachymetry, therapy by anti-glaucomatic drugs and regular inspections of the perimeter and HRT. The average post-surgery following-up time in our total of patients was 3 years and 8 months. The identified data of our total of patients were statistically processed using the Wilcoxon Signed-Rank Test. RESULTS: The average pre-surgery IOP was 21.4 mm Hg, 6 months post-surgery 11.2 mm Hg, and 13.2 mm Hg at the last inspection. Pre-surgery anti-glaucoma therapy: monotherapy in 2 eyes, dual therapy in 25 eyes, triple therapy in 13 eyes. Post-operatively without the need of therapy were 19 eyes, the need for monotherapy in 7 eyes, dual therapy in 13-eyes and triple therapy in 1 eye. During the last inspection of the perimeter or HRT within the postoperative period, we identified stationary finding in 39 eyes and only a mild progression in 1 eye. Peroperatively, we have not identified any serious complications. Within the postoperative period, we observed choroid ablation in 10 eyes as recovered within 6-7 days. As a more serious complication, we noted endophthalmitis in 1 eye on the background of generalized lichen planus complicating skin disease. In 5 eyes within the late postoperative period we found occlusion of lateral orifices in the EX-PRESS implant by the fibrotic tissue, the 4 eyes developed cataracts, and the eyeball hypotonia persists in 1 eye at the level of 5 torr without affecting the visus of the operated eye. CONCLUSION: It outflows from the above results that the use of EX-PRESS implant in the surgery of glaucoma is an effective and safe method with a minimal number of complications.

Tumor necrosis factor antagonism with etanercept improves systemic endothelial vasoreactivity in patients with advanced heart failure.

BACKGROUND: Anti-tumor necrosis factor (TNF)-alpha therapy with etanercept, a recombinant TNF receptor that binds to and functionally inactivates TNF-alpha, was shown to improve the functional status of patients with congestive heart failure (CHF). Because administration of TNF-alpha has been shown experimentally to depress endothelium-dependent relaxation, we hypothesized that TNF-alpha antagonism with etanercept might improve the depressed systemic endothelial vasodilator function, which importantly contributes to increased peripheral vascular resistance in patients with advanced CHF. METHODS AND RESULTS: Endothelium-dependent (acetylcholine, ACH; 10 to 50 microgram/min) and endothelium-independent (sodium nitroprusside, SNP; 2 to 8 microgram/min) forearm blood flow (FBF) responses were measured by venous occlusion plethysmography in 13 patients with documented CHF (New York Heart Association class III) before, 6 hours after, and 7 days after subcutaneous injection of a single dose of 25 mg etanercept. Maximum ACH-induced FBF increased significantly from 6.9+/-1.0 to 13.0+/-1.6 mL/min per 100 mL of forearm tissue (P<0.05) 6 hours after administration of etanercept and returned to 7.0+/-1.1 mL/min per 100 mL of forearm tissue after 7 days (P=NS), whereas SNP-induced FBF responses were not significantly affected. In contrast, FBF responses were not altered in control CHF patients, who did not receive etanercept (n=5). Etanercept-induced increases in ACH-mediated FBF were closely correlated with baseline TNF-alpha serum levels (r=0.66; P<0.02). CONCLUSIONS: The administration of etanercept profoundly improves systemic endothelial vasodilator capacity in patients with advanced heart failure, suggesting an important role of inflammatory mediators for impaired endothelial vasoreactivity in CHF. Improvement of systemic endothelial function might importantly contribute to the beneficial effects of etanercept on the functional status of patients with CHF.

Aging enhances the sensitivity of endothelial cells toward apoptotic stimuli: important role of nitric oxide.

Advanced aging leads to impaired endothelial NO synthesis and enhanced endothelial cell apoptosis; therefore, we investigated the sensitivity of aged endothelial cells toward apoptotic stimuli and determined the role of NO. Human umbilical vein endothelial cells (HUVECs) were cultured until 14th passage. In aged cells, oxLDL and tumor necrosis factor-alpha-induced apoptosis and caspase-3-like activity were significantly enhanced more than 3-fold compared with young cells (passage 3). Because NO contributes to protection against endothelial cell death via S-nitrosylation of caspases, we determined endothelial NO synthase (eNOS) protein expression and the content of S-nitrosylated proteins. Aged HUVECs showed significantly reduced eNOS expression (35+/-10%) and a decrease in the overall S-NO content (33+/-3%), suggesting that eNOS downregulation may be involved in age-dependent increase of apoptosis sensitivity. Indeed, eNOS knockout endothelial cells showed a significantly enhanced apoptosis induction. Exogenous NO donors abolished increased apoptosis and caspase-3-like activity. In contrast, the application of shear stress, which exerts a profound apoptosis inhibitory effect via upregulation of NO synthesis in young cells, failed to inhibit apoptosis in aged cells. Moreover, no upregulation of eNOS protein expression and S-NO content in response to shear stress was detected in aged cells. Overexpression of wild-type eNOS completely restored the antiapoptotic effect of shear stress, whereas only a partial inhibitory effect was detected under steady conditions. Strikingly, transfection of constitutively active phosphomimetic eNOS (S1177D) further abrogated apoptosis in aged HUVECs. Thus, aging of endothelial cells is associated with decreased NO synthesis and concomitantly increased sensitivity of apoptosis, which may contribute to functional impairment of the endothelial monolayer.

HMG-CoA reductase inhibitors (statins) increase endothelial progenitor cells via the PI 3-kinase/Akt pathway.

HMG-CoA reductase inhibitors (statins) have been developed as lipid-lowering drugs and are well established to reduce morbidity and mortality from coronary artery disease. Here we demonstrate that statins potently augment endothelial progenitor cell differentiation in mononuclear cells and CD34-positive hematopoietic stem cells isolated from peripheral blood. Moreover, treatment of mice with statins increased c-kit(+)/Sca-1(+)--positive hematopoietic stem cells in the bone marrow and further elevated the number of differentiated endothelial progenitor cells (EPCs). Statins induce EPC differentiation via the PI 3-kinase/Akt (PI3K/Akt) pathway as demonstrated by the inhibitory effect of pharmacological PI3K blockers or overexpression of a dominant negative Akt construct. Similarly, the potent angiogenic growth factor VEGF requires Akt to augment EPC numbers, suggesting an essential role for Akt in regulating hematopoietic progenitor cell differentiation. Given that statins are at least as potent as VEGF in increasing EPC differentiation, augmentation of circulating EPC might importantly contribute to the well-established beneficial effects of statins in patients with coronary artery disease.

Mutation of Trp1254 in the multispecific organic anion transporter, multidrug resistance protein 2 (MRP2) (ABCC2), alters substrate specificity and results in loss of methotrexate transport activity.

The ATP-binding cassette (ABC) proteins comprise a large superfamily of transmembrane transporters that utilize the energy of ATP hydrolysis to translocate their substrates across biological membranes. Multidrug resistance protein (MRP) 2 (ABCC2) belongs to subfamily C of the ABC superfamily and, when overexpressed in tumor cells, confers resistance to a wide variety of anticancer chemotherapeutic agents. MRP2 is also an active transporter of organic anions such as methotrexate (MTX), estradiol glucuronide (E217betaG), and leukotriene C4 and is located on the apical membrane of polarized cells including hepatocytes where it acts as a biliary transporter. We recently identified a highly conserved tryptophan residue in the related MRP1 that is critical for the substrate specificity of this protein. In the present study, we have examined the effect of replacing the analogous tryptophan residue at position 1254 of MRP2. We found that only nonconservative substitutions (Ala and Cys) of Trp1254 eliminated [3H]E217betaG transport by MRP2, whereas more conservative substitutions (Phe and Tyr) had no effect. In addition, only the most conservatively substituted mutant (W1254Y) transported [3H]leukotriene C4, whereas all other substitutions eliminated transport of this substrate. On the other hand, all substitutions of Trp1254 eliminated transport of [3H]MTX. Finally, we found that sulfinpyrazone stimulated [3H]E217betaG transport by wild-type MRP2 4-fold, whereas transport by the Trp1254 substituted mutants was enhanced 6-10-fold. In contrast, sulfinpyrazone failed to stimulate [3H]MTX transport by either wild-type MRP2 or the MRP2-Trp1254 mutants. Taken together, our results demonstrate that Trp1254 plays an important role in the ability of MRP2 to transport conjugated organic anions and identify this amino acid in the putative last transmembrane segment (TM17) of this ABC protein as being critical for transport of MTX.

Number and migratory activity of circulating endothelial progenitor cells inversely correlate with risk factors for coronary artery disease.

Recent studies provide increasing evidence that postnatal neovascularization involves bone marrow-derived circulating endothelial progenitor cells (EPCs). The regulation of EPCs in patients with coronary artery disease (CAD) is unclear at present. Therefore, we determined the number and functional activity of EPCs in 45 patients with CAD and 15 healthy volunteers. The numbers of isolated EPCs and circulating CD34/kinase insert domain receptor (KDR)-positive precursor cells were significantly reduced in patients with CAD by approximately 40% and 48%, respectively. To determine the influence of atherosclerotic risk factors, a risk factor score including age, sex, hypertension, diabetes, smoking, positive family history of CAD, and LDL cholesterol levels was used. The number of risk factors was significantly correlated with a reduction of EPC levels (R=-0.394, P=0.002) and CD34-/KDR-positive cells (R=-0.537, P<0.001). Analysis of the individual risk factors demonstrated that smokers had significantly reduced levels of EPCs (P<0.001) and CD34-/KDR-positive cells (P=0.003). Moreover, a positive family history of CAD was associated with reduced CD34-/KDR-positive cells (P=0.011). Most importantly, EPCs isolated from patients with CAD also revealed an impaired migratory response, which was inversely correlated with the number of risk factors (R=-0.484, P=0.002). By multivariate analysis, hypertension was identified as a major independent predictor for impaired EPC migration (P=0.043). The present study demonstrates that patients with CAD revealed reduced levels and functional impairment of EPCs, which correlated with risk factors for CAD. Given the important role of EPCs for neovascularization of ischemic tissue, the decrease of EPC numbers and activity may contribute to impaired vascularization in patients with CAD. The full text of this article is available at http://www.circresaha.org.

Increase in circulating endothelial progenitor cells by statin therapy in patients with stable coronary artery disease.

BACKGROUND: Therapeutic neovascularization may constitute an important strategy to salvage tissue from critical ischemia. Circulating bone marrow-derived endothelial progenitor cells (EPCs) were shown to augment the neovascularization of ischemic tissue. In addition to lipid-lowering activity, hydroxymethyl glutaryl coenzyme A reductase inhibitors (statins) reportedly promote the neovascularization of ischemic tissue in normocholesterolemic animals. Methods and Results-Fifteen patients with angiographically documented stable coronary artery disease (CAD) were prospectively treated with 40 mg of atorvastatin per day for 4 weeks. Before and weekly after the initiation of statin therapy, EPCs were isolated from peripheral blood and counted. In addition, the number of hematopoietic precursor cells positive for CD34, CD133, and CD34/kinase insert domain receptor was analyzed. Statin treatment of patients with stable CAD was associated with an approximately 1.5-fold increase in the number of circulating EPCs by 1 week after initiation of treatment; this was followed by sustained increased levels to approximately 3-fold throughout the 4-week study period. Moreover, the number of CD34/kinase insert domain receptor-positive hematopoietic progenitor cells was significantly augmented after 4 weeks of therapy. Atorvastatin treatment increased the further functional activity of EPCs, as assessed by their migratory capacity. CONCLUSION: The results of the present study define a novel mechanism of action of statin treatment in patients with stable CAD: the augmentation of circulating EPCs with enhanced functional activity. Given the well-established role of EPCs of participating in repair after ischemic injury, stimulation of EPCs by statins may contribute to the clinical benefit of statin therapy in patients with CAD.

Stabilization and cytoskeletal-association of LDL receptor mRNA are mediated by distinct domains in its 3' untranslated region.

The mRNA encoding the human low density lipoprotein (LDL) receptor is transiently stabilized after phorbol ester treatment of HepG2 cells and has been shown to associate with components of the cytoskeleton in this cell line (G. M. Wilson, E. A. Roberts, and R. G. Deeley, J. Lipid Res. 1997. 38: 437-446). Using an episomal expression system, fragments of the 3' untranslated region (3'UTR) of LDL receptor mRNA were transcribed in fusion with the coding region of beta-globin mRNA in HepG2 cells. Analyses of the decay kinetics of these beta-globin-LDL receptor fusion mRNA deletion mutants showed that sequences in the proximal 3'UTR of LDL receptor mRNA including several AU-rich elements (AREs) were sufficient to confer short constitutive mRNA half-life in the heterologous system. Stabilization of LDL receptor mRNA in the presence of PMA required sequences in the distal 3'UTR, at or near three Alu-like repetitive elements. Furthermore, the 3'UTR of LDL receptor mRNA conferred cytoskeletal association on the otherwise unassociated beta-globin mRNA, by a mechanism involving at least two distinct RNA elements. Comparisons of decay kinetics and subcellular localization of endogenous LDL receptor mRNA and beta-globin-LDL receptor mRNA fusions in HepG2 cells have demonstrated that several cis-acting elements in the receptor 3'UTR contribute to post-transcriptional regulation of receptor expression, and provide further support for involvement of the cytoskeleton in the regulation of LDL receptor mRNA turnover.

[Modulation of wound healing in glaucoma filtration surgery]. / Modulace hojení rány pri antiglaukomových filtrujících operacích.

Authors evaluate application of mitomycin C (0.2-0.4 mg/ml) on the sclera during glaucoma filtering surgery. Mean follow up was 19.5 months in the group of 67 patients (75 eyes). The most serious complication was an early endophthalmitis in 2 cases with the loss of residual visual field and vision, 1x hemophthalmus followed with development of traction retinal detachment, 1x hypotonic maculopathy, 1x traction retinal detachment for progression of diabetic proliferative vitreoretinopathy of neovascular glaucoma. Higher rate of complications was noted in the group of mitomycin C 0.4 mg/ml. Visual acuity was better one line Snellen optotypes in 3 cases, and worse in 26 cases. Intraocular pressure compensation up to 20 mmHg was reached in 32 cases without therapy (45.1%), in 28 cases therapy (39.6%), Intraocular pressure above 20 mm Hg was in 11 cases (15.4) and the data are not known in 4 cases (5.4%). Intraocular pressure was compensated after mitomycin C application in 84.7%.

[Mitomycin C in glaucoma filtering surgery]. / Vyuzití mitomycinu C pri filtrujících glaukomových operacích.

Authors evaluate application of mitomycin C (0.2-0.4 mg/ml) on the sclera during glaucoma filtering surgery. Mean follow-up was 19.5 months in the group of 67 patients (75 eyes). The most serious complication was an early endophthalmitis in 2 cases with the loss of residual visual field and vision, 1x hemophthalmus followed with the development of traction retinal detachment, 1x hypotonic maculopathy, 1x traction retinal detachment for progression of proliferative vitreoretinopathy of neovascular glaucoma. Higher rate of the complications was noted in the group of mitomycin C 0.4 mg/l. Visual acuity was better one line Snellen optotypes in 3 cases, and worse in 26 cases. Intraocular pressure (IOP) compensation up to 20 mmHg was reached in 32 cases without therapy (45.1%), in 28 cases therapy, (39.6%). IOP above 20 mmHg was in 11 cases (15.4%) and the data are not known in 4 cases (5.41%). IOP was compensated after mitomycin C application in 84.7%.

[Refractive lensectomy with intraocular lens implantation in hypermetropia gravis]. / Refrakcní lensektomie s implantací nitroocní cocky u hypermetropia gravis.

The objective of the study was to evaluate the results of refractive lensectomy with implantation of an IOL in patients with hypermetropia gravis. The authors included in the group patients operated at the Ophthalmological Clinic in Hradec Králové between May 1995 and April 1997 with a refraction of +7.2 +/- 1.76 D and axial length of 19.91 +/- 1.23 mm. The mean value of IOL was 32 +/- 1.8 (+30.0 to +35.0 D). The group comprised 124 men and 23 women (range 21-67 years, mean age 45.8 +/- 10.5 years). From the total number of 94 eyes 22 were operated, while the patients were hospitalized, 72 were operated at the out-patient department. Visus of 6/12 and better with supplementary correction was achieved in 73 eyes, i.e. 77% during the last check up. Before operation thist was the case in 70 eyes, i.e. 75.3%. The mean refraction after operations was -0.25 +/- 1.05 D. Lensectomy in hypermetropia gravis with implantation of an intraocular lens is according to the achieved functional results a fully justified method for resolving the problem of severe far-sightedness.

cIRF-3, a new member of the interferon regulatory factor (IRF) family that is rapidly and transiently induced by dsRNA.

In mammals, some of the effects of interferon (IFN) on gene transcription are known to be mediated by a family of IFN-inducible DNA-binding proteins, the IFN regulatory factor (IRF) family, which includes both activators and repressors of transcription. Although IFN activities have been described in many vertebrates, little is known about regulation of IFN- or IFN-stimulated genes in species other than human and mouse. Here, we report the cloning of a chicken cDNA, cIRF-3, encoding a protein with a DNA-binding domain similar to that found in the mammalian IRF family of proteins. Similarity between cIRF-3 and the mammalian IRFs is comparable with that between known members of the family. It is most similar to the IRF proteins ICSBP and ISGF3 gamma but is equally divergent from both. Gel mobility shift assays indicate that cIRF-3 is capable of binding a known IFN-stimulated response element that is conserved between the mammalian and chicken Mx genes. Expression of the cIRF-3 gene can be induced to high levels by poly(I).poly(C). Induction is rapid and transient with no requirement for protein synthesis. Co-treatment of cells with cycloheximide results in superinduction of cIRF-3 mRNA. The structural and regulatory characteristics of cIRF-3 indicate that it is the first example of a non-mammalian IRF protein.

Developmental regulation of specific protein interactions with an enhancerlike binding site far upstream from the avian very-low-density apolipoprotein II gene.

Expression of the avian very-low-density apolipoprotein II (apoVLDLII) gene is completely dependent on estrogen and restricted to the liver. We have identified binding sites for nonhistone nuclear proteins located between -1.96 and -2.61 kilobases. One of these sites, located at -2.6 kilobases (designated site 1), was found to span an MspI site that becomes demethylated between days 7 and 9 of embryogenesis, the stage of development at which competence to express the apoVLDLII gene begins to be acquired. Levels of the factor(s) involved were high at day 7 of embryogenesis, decreased two- to threefold by days 9 to 11, and continued to decline more slowly until hatching. Furthermore, the mobility of the complex formed underwent a well-defined shift between days 11 to 13 embryogenesis. Methylation interference studies showed that modification of the outer guanosines of the MspI site resulted in marked inhibition of the formation of the protein-DNA complex. Competition studies, fractionation of nuclear extracts, and tissue distribution indicated that the factor was not the avian homolog of hepatocyte nuclear factor 1, nuclear factor 1, or CCAAT/enhancer-binding protein (C/EBP). However, site 1 could complete for binding to an oligonucleotide, previously shown to be recognized by C/EBP, in a nonreciprocal fashion. These studies demonstrate that the sequence recognized by the protein includes a C/EBP consensus sequence but that elements in addition to the core enhancer motif are essential for binding.

Epidemiological aspects of Salmonella infantis and Salmonella typhi murium infections in Finnish broiler chickens.

In this paper salmonella infections in Finnish broilers have been described. Especially infections detected in the farms of company A have been studied. Attention has been paid to the year by year continuing numerous S. infantis infections. In agreement with earlier reports, there is some evidence suggesting that a hatchery might have spread the infections. The examination of grandparent and parent birds as well as hatcheries with respect to salmonella infections should be more thorough and clear. It is, for example, for the moment not known at the farms of company A from which parent bird flocks the chickens come. This knowledge should be very important when the origin of salmonella infections is tried to be clarified.

The 5' noncoding and flanking regions of the avian very low density apolipoprotein II and serum albumin genes. Homologies with the egg white protein genes.

We have isolated and sequenced the 5' proximal exons and flanking regions of the chicken very low density apolipoprotein II (apo-VLDLII) and serum albumin genes. These genes specify the most abundant mRNA species present in livers of hens or estrogen-treated roosters. Sequencing revealed that the promoter region of the estrogen-dependent apo-VLDLII gene contained at least two potential transcriptional initiation sites, preceded by appropriately positioned "ATA" sequences. One is homologous with the cap site of the ovalbumin gene, and the other exhibits a match of 10 out of 12 nucleotides with the cap site of the serum albumin gene. S1 nuclease mapping indicates that both sites are active in vivo, although the "ovalbumin"-like site is by far the most efficient at all stages of the estrogenic response. The relative efficiencies of these two sites are maintained during in vitro transcription of truncated templates. A third site, that was not anticipated from sequencing data, is active in vivo but inactive in vitro. A conserved 5' flanking element, initially identified during studies on egg white protein genes, is also present upstream from the apo-VLDLII and serum albumin genes. Its removal does not affect initiation site selection in vitro. Regions of the apo-VLDLII and ovalbumin genes extending 100 nucleotides downstream from the "TATA box" contain several striking homologies that suggest a common mode of evolution.