Preservation of the Photoreceptor Inner/Outer Segment Junction in Dry Age-Related Macular Degeneration Treated by Rheohemapheresis.

Aim. To evaluate the long-term effect of rheohemapheresis (RHF) treatment of age-related macular degeneration (AMD) on photoreceptor IS/OS junction status. Methods. In our study, we followed 24 patients with dry AMD and drusenoid retinal pigment epithelium detachment (DPED) for a period of more than 2.5 years. Twelve patients (22 eyes) were treated by RHF and 12 controls (18 eyes) were randomized. The treated group underwent 8 RHF standardized procedures. We evaluated best-corrected visual acuity, IS/OS junction status (SD OCT), and macular function (multifocal electroretinography) at baseline and at 2.5-year follow-up. Results. RHF caused a decrease of whole-blood viscosity/plasma viscosity at about 15/12%. BCVA of treated patients increased insignificantly (P = 0.187) from median 74.0 letters (56.2 to 81.3 letters) to median 79.0 letters (57.3 to 83.4 letters), but it decreased significantly from 74.0 letters (25.2 to 82.6 letters) to 72.5 letters (23.4 to 83.1 letters) in the control group (P = 0.041). The mfERG responses in the region of eccentricity between 1.8° and 7° were significantly higher in treated patients (P = 0.04). Conclusions. RHF contributed to sparing of photoreceptor IS/OS junction integrity in the fovea, which is assumed to be a predictive factor for preservation of visual acuity.

Transient branch retinal artery occlusion in a 15-year-old girl and review of the literature.

BACKGROUND: Retinal artery occlusion is an extremely rare diagnosis in the pediatric population and the etiology with risk factors of retinal artery occlusion are poorly understood in younger individuals. METHODS AND RESULTS: This case report a rare case of transient branch retinal artery occlusion (BRAO) in a healthy young girl. A fifteen-year-old girl presented with painless decreased vision in her right eye. Fundus examination revealed in the right eye intensive edema of the part of superior half of the retina, but the vessels were absolutely normal. She was hospitalized for diagnosis and treatment. A thorough workup was performed to determine any etiologic factor. Her physical examination was normal, but from anamnesis has been found that patient began use oral contraception at the time of occurrence eye problems. The condition is evaluated as transient branch retinal artery occlusion by the course of the disease. The paper includes the review of the literature also. CONCLUSION: Though retinal arterial obstruction is rare in the pediatric population, this case highlights the importance of including this in the differential diagnosis of acute vision loss.

Dynamics of blood count after rheohemapheresis in age-related macular degeneration: possible association with clinical changes.

BACKGROUND: Rheohemapheresis (RHF) is a method that can stop the activity of the dry form of age-related macular degeneration (AMD). The pathophysiologic mechanisms are not well understood, and the effects of the RHF procedures extend beyond the time of the individual procedures. PATIENTS AND METHODS: We present the data for 46 patients with AMD treated with a series of 8 rheohemapheretic procedures. Blood count parameters were measured before the first and the last procedures. The clinical effect was judged by changes in the drusenoid pigment epithelium detachment (DPED) area before and after the rheopheretic sessions. RESULTS: Rheopheresis caused a decrease in hemoglobin (P<0.001), a decrease in leukocytes (P<0.034), and an increase in platelets (P<0.005). We found a negative correlation between the amount of platelets and their volume (P<0.001, Pearson correlation coefficient: -0.509). We identified the platelet/MPV ratio as a good predictor of the clinical outcome. Patients with a platelet/MPV ratio greater than 21.5 (before the last rheopheresis) had a significantly better outcome (P=0.003, sensitivity of 76.9% and specificity of 80%). CONCLUSION: Several basic blood count parameters after RHF can be concluded to significantly change, with some of those changes correlating with the clinical results (reduction of the DPED area).

Cost and effectiveness of therapy for wet age-related macular degeneration in routine clinical practice.

PURPOSE: Evaluation of the cost and effectiveness of therapy for patients with the wet form of age-related macular degeneration (AMD) in routine clinical practice. METHODS: A retrospective multicentre evaluation of changes in the best-corrected visual acuity in applied kinds of therapy and a comparison with the cost of individual therapeutic procedures. RESULTS: An overall total of 788 eyes of 763 patients with an average age of 73.2 ± 8.6 years was evaluated for a 1-year minimum period. In the ranibizumab and pegaptanib therapy groups, a reduction of 1.3 letters (p = 0.303) and 1.4 letters (p = 0.197) was found, respectively. In the group of photodynamic therapy (PDT) with verteporfin, a reduction of 5.2 letters was achieved (p < 0.001). Under the conditions of routine practice in the Czech Republic, the annual cost is highest (EUR 5,467.63/patient) in patients with pegaptanib therapy. The annual cost in patients with ranibizumab therapy is lower by EUR 1,220.16. The cost is nearly half (EUR 2,783.65) in the group treated with PDT with verteporfin. CONCLUSION: An initiation of AMD therapy by ranibizumab is cost-effective as compared to pegaptanib. Both ranibizumab and pegaptanib are significantly more efficient as compared to PDT with verteporfin. Therapy with ranibizumab and pegaptanib, as compared to PDT with verteporfin, prevents the loss of 1 line of vision on the ETDRS chart for EUR 1,225.98 and 2,286.18, respectively.

Long-term outcomes of rheohaemapheresis in the treatment of dry form of age-related macular degeneration.

Purpose. Determining long-term effects of rheohaemapheresis on the dry form of age-related macular degeneration. Methods. This study evaluates 19 patients, average age of 67.6 years, treated with rheohaemapheresis and 18 patients, average age of 72.8 years, comprising the control group. Minimum follow up period was 3.5 years. Each treated patient received a series of 8 sessions of rheohaemapheresis of 1.5 plasma volumes within 10 weeks. We measured the drusenoid pigment epithelium detachment (DPED), best-corrected visual acuity (BCVA), electroretinography (ERG), and rheological parameters. Results. In the treatment group, the baseline BCVA was 0.74 (0.36-1.0) 95% CI and BCVA after 3.5 years was 0.79 (0.41-1.0) 95% CI (P = 0.726). In the control group, the baseline BCVA was 0.71 (0.15-1.0) 95% CI and BCVA after 3.5 years decreased to 0.7 (0.32-0.87) 95% CI (P = 0.031). Baseline DPED was 6.78 ± 3.79 mm(2); after 3.5 years, it decreased to 4.13 ± 3.84 mm(2) (P < 0.001). In the control group, the baseline DPED was 4.09 ± 3.48 mm(2); after 3.5 years, it increased to 6.69 ± 4.2 mm(2) (P = 0.001). We noted increasing levels of positive wave peaking at 50 milliseconds (P50) after treatment (P = 0.022) and a stable amplitude of photopic responses of treated patients. Conclusion. Over the long term, rheohaemapheresis reduced the DPED, improved the function of photoreceptors, and prevented the decline of BCVA.

Effects of treatment change in patients with neovascular age-related macular degeneration; results from the Czech National Registry.

AIMS: To determine the effectiveness of second line treatments in patients with neovascular AMD who did not respond adequately to primary treatment. METHODS: Retrospective, multicentre assessment. The frequency of primary treatment failure and outcomes of subsequent secondary treatment were assessed according to the type of primary treatment, type of CNV and change in BCVA over a 12 month period. RESULTS: At the time of assessment 750 entries (750 treated eyes, 725 treated patients) had follow-up longer than 12 months. A treatment change required 7.7% subjects treated with ranibizumab, 20.5% with pegaptanib and 22% with PDT and verteporfin. Average BCVA of all patients at the beginning of primary treatment was 50.7 ± 3 letters and 43 ± 3.5 letters in 12(th) month (P<0.001). The mean decrease in BCVA was 7.7 ± 0.6 letters during the first 6 months of observation. During the next 6 months, no significant change occurred. The change of primary therapy was required on average after 6.5 ± 2.1 months. CONCLUSION: BCVA loss was the most significantly decelerated in patients who received ranibizumab as a secondary therapy following unsuccessful treatment with pegaptanib sodium.

The importance of rheological parameters in the therapy of the dry form of age-related macular degeneration with rheohaemapheresis.

To date, rheological treatment is the only chance to control the advanced dry form of age-related macular degeneration and arrest its progression to legal blindness. Rheohaemapheresis can change the main rheological parameters, blood and plasma viscosity, as well as change erythrocyte aggregability, improve erythrocyte flexibility and lead to substantial improvement when other methods of therapy fail. In this study, we describe changes in the levels of rheological efficacy indicators after rheohaemapheresis and their clinical significance in the dry form of age-related macular degeneration (AMD). Seventy-two patients with AMD were randomised; 34 controls, and 38 patients were treated with rheohaemapheresis (separator Cobe Spectra + Evaflux filter). After the procedures, α2-macroglobulin levels decreased by approximately 58%, fibrinogen by approximately 65%, IgM by approximately 67%, LDL cholesterol by approximately 71%, apolipoprotein B by approximately 65%, and lipoprotein (a) by approximately 42%. These decreases correspond with a decrease in blood and plasma viscosity (14/12%), clinical improvement (arrest of disease progression, even visual improvement in some cases), and heretofore-unreported improvement (even reattachment) of drusen retinal pigment epithelium detachment. Our modification of rheohaemapheresis is safe (5.4% of patients experienced clinically insignificant side effects).

One-step multiplex PCR method for the determination of pecan and Brazil nut allergens in food products.

BACKGROUND: A one-step polymerase chain reaction (PCR) method for the simultaneous detection of the major allergens of pecan and Brazil nuts was developed. Primer pairs for the amplification of partial sequences of genes encoding the allergens were designed and tested for their specificity on a range of food components. RESULTS: The targeted amplicon size was 173 bp of Ber e 1 gene of Brazil nuts and 72 bp of vicilin-like seed storage protein gene in pecan nuts. The primer pair detecting the noncoding region of the chloroplast DNA was used as the internal control of amplification. The intrinsic detection limit of the PCR method was 100 pg mL(-1) pecan or Brazil nuts DNA. The practical detection limit was 0.1% w/w (1 g kg(-1)). The method was applied for the investigation of 63 samples with the declaration of pecans, Brazil nuts, other different nut species or nuts generally. In 15 food samples pecans and Brazil nuts allergens were identified in the conformity with the food declaration. CONCLUSION: The presented multiplex PCR method is specific enough and can be used as a fast approach for the detection of major allergens of pecan or Brazil nuts in food.

Haemorheopheresis could block the progression of the dry form of age-related macular degeneration with soft drusen to the neovascular form.

PURPOSE: To evaluate the influence of haemorheopheresis on anatomical and functional findings in patients with soft-drusen maculopathy. METHODS: We investigated 29 eyes (16 patients) and randomized 25 eyes (16 controls) with soft-drusen maculopathy [soft, confluent and reticular drusen, drusenoid retinal pigment epithelium detachment (RPED)]. Each patient received a series of eight haemorheophereses (cascade filtration of 1.5 plasma volume) within 10 weeks. The patients were followed up using Early Treatment Diabetic Retinopathy Study (ETDRS) charts, optical coherence tomography, fluorescein angiography, electroretinography and measurements of pulsed ocular blood flow. RESULTS: After the procedures, there was a substantial reduction in rheologically active substances [lipoproteins, α2-macroglobulin, immunoglobulin M (IgM), fibrinogen], plasma and blood viscosity. At the 1.5-year follow-up, we noticed soft drusen absorption; reattachment of drusenoid RPED and stabilization or improvement of visual acuity occurred in 72% of patients in comparison to only 39% of patients in the control group. Full-field electroretinograms showed significantly higher scotopic activity of treated patients in comparison with the control group, and mainly insignificant differences in photopic activity between both groups. Despite the significant increase of activity in the paramacular retina in treated patients, the differences in amplitudes of multifocal electroretinography (mfERG) average responses were insignificant between groups. CONCLUSION: Haemorheopheresis seems to be capable of changing the activity of promoters of the natural course of soft-drusen maculopathy, its development and progression. Visual acuity and electrical activity of the retina can be stabilized or even improved. The therapy has been shown to be effective and safe.

Visual functions after laser photocoagulation in central serous chorioretinopathy.

The authors evalute the visual functions of patients with unilateral central serous chorioretinopathy (CSC) and compare them with their non-affected eye and with a control group. Fourteen patients with CSC treated with direct laser-photocoagulation were examined preoperatively and followed-up, up to 2 years postoperatively. Baseline best corrected visual acuity (BCVA) and contrast sensitivity (CS) of both eyes of patients was significantly lower in comparison with the controls. BCVA and CS in affected eyes were significantly lower compared to the fellow eye of patients. The final BCVA and CS of patients did not differ significantly from the controls, except CS of affected eyes in the spatial frequency of 3.69 c/deg. Two years after laser treatment, there were only nonsignificant differences of both photopic full-field electroretinography (phERG) and multifocal electroretinography (mfERG) responses between the treated, the nonaffected eye of the patients and the control group with exception of a significantly longer P1 implicit time in the parafoveolar region in affected eyes. Colour discrimination was normal in 85.8% of affected eyes of the patients. Despite a significant improvement of macular function in CSC eyes, functional examination methods do not prove complete resolution of function 2 years after laser-treatment.

Identification of gadoid species in fish meat by polymerase chain reaction (PCR) on genomic DNA.

Identification of fish species is significant due to the increasing interest of consumers in the meat of sea fish. Methods focusing on fish species identification help to reveal fraudulent substitution among economically important gadoid species in commercial seafood products. The objective of this work was to develop a conventional PCR method for the differentiation of the following gadoid fish species in fish products: Alaska pollack ( Theragra chalcogramma), blue whiting ( Micromesistius poutassou), hake spp. ( Merluccius spp.), Atlantic cod ( Gadus morhua), saithe ( Pollachius virens), and whiting ( Merlangius merlangus). The species-specific primer pairs for gadoid species determination were based on the partial pantophysin I ( PanI) genomic sequence. Sequence identification was confirmed by cloning and sequencing of the PCR products obtained from the species considered. For the simultaneous detection of Alaska pollack, blue whiting, and hake spp., a quadruplex PCR system was constructed. Other gadoid species were detected in separate PCR reactions. After optimization of the reactions, the developed PCR systems were used for the analysis of codfish samples obtained from the Czech market and the customs' laboratories. This method represents an alternative approach in the use of genomic DNA for the identification of fish species. This method is rapid, simple, and reliable without the need for further confirmative methods. Furthermore, the identification of a mixture of more than one species is possible. The PCR system has been optimized for routine diagnostic purposes.

Quantitative detection of species-specific DNA in feedstuffs and fish meals.

A sensitive and rapid method for the quantitative detection of bovine-, ovine-, swine-, and chicken-specific mitochondrial DNA sequences based on real-time PCR has been developed. The specificity of the primers and probes for real-time PCR has been tested using DNA samples of other vertebrate species that may also be present in rendered products. The quantitative detection was performed with dual-labeled probes (TaqMan) using absolute quantification with external standards of single species meat-and-bone meals. This method facilitates the detection of 0.01% of the target species-derived material in concentrate feed mixtures and fish meals.

Comparison of commercially available antibodies for the detection of central nervous system tissue in meat products by enzyme-linked immunosorbent assay.

An indirect competitive enzyme-linked immunosorbent assay was used to compare four commercially available monoclonal and polyclonal antibodies prepared against the markers of central nervous system tissue: glial fibrillary acidic protein, neurofilaments, and myelin basic protein. These antibodies were investigated for their usefulness at the detection of prohibited central nervous system tissue in meat products to prevent spread of bovine spongiform encephalopathy and its human form, a new variant of Creutzfeldt-Jakob disease. The detection limit was 2.5 to 10%. The specificity was tested using extracts of tissue samples of bovine and porcine brain, spinal cord, lymphatic node, diaphragm, skeletal muscle, and peripheral nerves. All antibodies reacted with both bovine and porcine tissue extracts.

[Results of transpupillary thermotherapy by patients with age-related macular degeneration 6 months after treatment (pilot study)]. / Výsledky transpupilární termoterapie u pacientu s vekem podmínenou makulární degenerací 6 mesícu po lécbe (pilotní studie).

We demonstrated the effectiveness of transpupillary thermotherapy (TTT) for the treatment of choroidal neovascularization (CNV) by patients with age- related macular degeneration (ARMD). CNV secondary to ARMD is a leading case of vision loss in patients older than 50 years. TTT is new promising method for the treatment CNV by ARMD. It is able to close CNV while maintaining visual function and in small percentage visual function improving. We have performed TTT by 30 patients (30 eyes) with subfoveolar CNV. We have demonstrated the stabilization or improvement of visual acuity by 16 patients (52%), improvemend two or more lines was documented by 5 patients (17%).

[Age-related macular degeneration--contemporary possibilities of treatment]. / Vekem podmínená makulární degenerace sítnice--soucasné moznosti lécby.

Age-related macular degeneration (AMD) is a complex multifactorial disease, that affects the central region of the retina. AMD is a leading cause of blindness among people over 50 years of age in the industrialized world. Two types of macular degeneration exist, the "dry" form and the "wet-neovascular" form. Neovascular AMD is responsible for 80-90% of cases of severe, irreversible central vision loss in patients over 65 years of age with AMD. The authors present contemporary possibilities of treatment of the patients with this disease.

Identification of species-specific DNA in feedstuffs.

Due to the menace of transmission of spongiform encephalopathies, feed components intended for ruminant nutrition must be checked for the presence of ruminant-derived materials. A sensitive method for the identification of bovine- and ovine- and also swine- and chicken-specific mitochondrial DNA sequences based on Polymerase Chain Reaction (PCR) has been developed. The specificity of the primers for PCR has been tested using samples of DNA of other vertebrate species, which may also be present in rendering plant products intended for feed manufacture. The method allows the detection in concentrate mixtures of 0.01% of the target species derived material. The identity of a sample containing 0.1% of bovine, ovine, swine, and chicken meat-and-bone meal has further been confirmed by sequencing.