Recommendations for the Management of Uveitis Associated With Juvenile Idiopathic Arthritis: The Czech and Slovak adaptation of SHARE Initiative.

Juvenile idiopathic arthritis (JIA) is the most common rheumatic disease in children and uveitis is its most important extra-articular manifestation. Evidence-based recommendations are available only to a limited extent and therefore JIA associated uveitis management is mostly based on physicians experience. Consequently, treatment practices differ widely, both nationally and internationally. Therefore, an effort to optimize and publish recommendations for the care of children and young adults with rheumatic diseases was launched in 2012 as part of the international project SHARE (Single Hub and Access Point for Pediatric Rheumatology in Europe) to facilitate clinical practice for paediatricians and (paediatric) rheumatologists. The aim of this work was to translate published international SHARE recommendations for the diagnosis and treatment of JIA associated uveitis and to adapt them for use in the Czech and Slovak Republics. International recommendations were developed according to the standard methodology of the European League against Rheumatism (EULAR) by a group of nine experienced paediatric rheumatologists and three experts in ophthalmology. It was based on a systematic literature review and evaluated in the form of an online survey and subsequently discussed using a nominal group technique. Recommendations were accepted if > 80% agreement was reached (including all three ophthalmologists). A total of 22 SHARE recommendations were accepted: 3 on diagnosis, 5 on disease activity assessment, 12 on treatment and 2 on future recommendations. Translation of the original text was updated and modified with data specific to the czech and slovak health care systems and supplemented with a proposal for a protocol of ophthalmological dispensarization of paediatric JIA patients and a treatment algorithm for JIA associated uveitis. Conclusion: The aim of the SHARE initiative is to improve and standardize care for paediatric patients with rheumatic diseases across Europe. Therefore, recommendations for the diagnosis and treatment of JIA-associated uveitis have been formulated based on the evidence and agreement of leading European experts in this field.

Identification and characterization of a new member of the genus Luteovirus from cherry.

The complete genomic sequence of a new virus from cherry trees was determined. Its genome is 5857 nt long and resembles that of members of the genus Luteovirus in its genomic organization and nucleotide sequence. Based on the species demarcation criteria for luteoviruses, the virus represents a new luteovirus species. Furthermore, a 47-nt-long inverted repeat was found at the 3' end of its genome. The virus has been provisionally named cherry-associated luteovirus (ChALV) and is the fourth member of the family Luteoviridae reported to naturally infect woody plants.

Methotrexate efficacy, but not its intolerance, is associated with the dose and route of administration.

BACKGROUND: There is a lack of published evidence on the importance of methotrexate (MTX) dose and route of administration on both its efficacy and adverse events in children with Juvenile Idiopathic Arthritis (JIA). We aimed to document our clinical practice based on the treat-to-target approach in order to support the concept that better therapeutic effect achieved with an optimal dose of parenteral MTX is associated with clinically acceptable adverse effects comparable to those reported for oral treatment. METHODS: Study inclusion criteria were indication of new MTX therapy for active arthritis in confirmed JIA patients younger than 18 years. Eligible patients were evaluated prospectively every 3 months for 1 year using standardized instruments for treatment response (American College of Rheumatology Pediatric (ACRPedi) response, Juvenile Arthritis Disease Activity Score (JADAS) 71, Clinically Inactive Disease (CID)) and adverse events (laboratory monitoring, Methotrexate Intolerance Severity Score (MISS)). MTX responders had to achieve at least ACRPedi 70 response. MTX intolerance was defined by MISS ≥ 6. RESULTS: In 45/55 patients (81.8 %) MTX was started as subcutaneous injection. The initial median weekly dose was 14.4 mg/m(2) in parenteral and 11.7 mg/m(2) in oral administration. MTX therapy was effective in the level of ACRpedi70 and CID in 50.9 % and 30.9 % of patients at month 6 and in 70.9 % and 56.4 % after 12 months of the treatment, respectively. MTX intolerance at 6 and 12 months was noted in 25.5 % and 30.6 %, respectively. Management of intolerance included change in the dose and/or route of administration, education and councelling. Adverse events led to MTX withdrawal in 5 patients (9 %) due to toxicity (n = 3) and intolerance (n = 2). We did not find any significant predictive factors for either MTX therapeutic response or intolerance. CONCLUSION: Subcutaneous MTX weekly dose around 15 mg/m(2) is associated not only with a high response rate within the first 12 months of treatment, but also with a relatively low rate of significant adverse effects that would lead to the treatment termination. It allows early recognition of MTX non-responders and addition of biologic therapy. Sustainability of therapeutic effect and longer-term evolution of adverse events will be addressed by an ongoing extension of the study.

Association of 'Candidatus Phytoplasma asteris' with yellowing and phyllody of Plantago lanceolata.

Long plantains (Plantago lanceolata L.) with symptoms resembling those associated with phytoplasma infection were observed repeatedly during the period 2000-2008 in southern Bohemia (Czech Republic). The symptoms of the plants were leaf yellowing, stunted growth, flower phyllody and lack of seed production. Transmission electron microscopy showed phytoplasmas in the sieve cells of affected plants but not in healthy ones. Association of phytoplasmas with the disease was confirmed by polymerase chain reaction using phytoplasma-specific universal ribosomal primers R16F2n/R16R2. An amplification product of the expected size (1.2 kb) was observed in all samples of the symptomatic long plantains. The restriction profiles obtained from digestion of the PCR products with three endonucleases (AluI, HhaI, MseI) showed that the phytoplasmas infecting long plantains in the Czech Republic were indistinguishable from those belonging to the aster yellows group (subgroup 16SrI-B). Sequence analysis of 1748 bp of the ribosomal operon indicated that the closest related phytoplasma was that associated with 'Rehmannia glutinosa var. purpurea', originating also in Bohemia. This is the first report of the natural occurrence of 'Candidatus Phytoplasma asteris' in plants of P. lanceolata.

Phytoplasma associated with witches'-broom disease of Ulmus minor MILL . in the Czech Republic: Electron microscopy and molecular characterization.

Visual inspections of elm trees in south Moravia in 1997-2007 revealed a rare occurrence of plants with smaller and cowl-forming leaves on some twigs, i.e. a feature resembling witches'-broom disease observed on the end of twigs. The presence of phytoplasma-like bodies was observed by transmission electron microscopy of phloem tissue. On the other hand, no phytoplasmas were found in asymptomatic trees. Nucleic acids extracted from these plants were used in nested-PCR assays with primers amplifying 16S rRNA sequences specific for phytoplasmas. Sequence analyses of the 16S-23S ribosomal operon (1852 bp) allowed for the classification of the detected phytoplasmas in the elm yellows group, but its position remained on the boundary of the 16SrV-A and 16SrV-C ribosomal subgroups. Sequence analyses of the ribosomal protein of the rpl22-rps3 and secY genes lead to further classification and revealed the phytoplasmas' affiliations to the 'Candidates Phytoplasma ulmi'. Some exceptions in unique oligonucleotide sequences defined for 'Ca. Phytoplasma ulmi' were found in the Czech isolate. This is the northernmost confirmed occurrence of phytoplasma on elm trees within Europe.

[Contribution of allergen immunotherapy using Phostal in the treatment of seasonal allergic rhinitis (two years of use)]. / Prínos alergenové imunoterapie Phostalem v lécbe sezonní alergické rýmy (dva roky aplikace).

Allergen immunotherapy consists in the administration of the appropriate allergen to which the patient is hypersensitive. The objective is to induce immunological tolerance of the organism. In the present study, the effect of the administration of Phostal on the occurrence of seasonal symptoms of pollinosis in a group of 35 outpatients was observed during two years. The substance was administered according to the regular dosage schedule, all year round. The results of the study have demonstrated a significant effect of the product on the decrease in the occurrence of observed objective symptoms of the disease, starting already from the first year of treatment. This is in correlation with changes in the spirometric functions, selected laboratory parameters and results of the prick tests in the treated group in comparison to the control group.

[Testing of antidiabetic and antioxidative effect of the flavonoid osajin in an experiment]. / Testování antidiabetického a antioxidacního úcinku flavonoidu osajinu v experimentu.

The antidiabetic and antioxidative effect of osajin was monitored under the conditions of alloxan-induced diabetes mellitus in an in vivo experiment. The animals were divided by random selection into 2 groups (n=7). The treated group was administered osajin in peroral doses of 10 mg/kg in Avicel, the placebo diabetic group was given only the solution of Avicel, and the last group was intact. Selected laboratory parameters (glucose, urea, cholesterol, antioxidative enzymes, total antioxidative capacity, malondialdehyde in serum, diuresis, total glucose and protein losses through urine) were determined in all animals. Kidney tissue and pancreas samples were taken for histopathological analysis. The findings included a statistically significant increase (p< or =0.05) in the glutathione peroxidase catalytic activity, total antioxidative capacity (p< or =0.01) and a statistically significant decrease (p< or =0.01) on malondialdehyde level in the treated group compared to the placebo group. A statistically highly significant decrease (p< or =0.01) in diuresis, glucose and protein losses through urine were identified in the treated group compared to the placebo group. The superoxide dismutase catalytic activity, urea and cholesterol levels involved non-significant changes. The results of biochemical examination show a protective antidiabetic and antioxidative effect of osajin. The results of histopathological examination correlate with them only partially.

[Examination of the antioxidative and antidiabetic effect of pomiferin in alloxan-induced diabetes mellitus in an experiment (a pilot study)]. / Sledování antioxidacního a antidiabetického efektu pomiferinu u alloxanem navozeného diabetes mellitus v experimentu--pilotní studie.

The antioxidative and antidiabetic effect of pomiferin was monitored under the experimental conditions of alloxan-induced diabetes mellitus. The animals were divided by random selection into 2 groups (n=7). The treated group was administered pomiferin in peroral doses of 10 mg/kg in Avicel, the placebo group was given only a solution of Avicel, and the last group was intact. Selected laboratory parameters (glucose, urea, cholesterol, antioxidative enzymes, total antioxidative capacity, malondialdehyde in serum: diuresis, total glucose and protein losses through urine) were determined. Kidney tissue and pancreas samples were taken for histopathological analysis. The findings included a statistically significant decrease (p < or = 0.01) in blood glucose level, a significant increase (p < or = 0.01) in glutathione peroxidase, total antioxidative capacity (p < or = 0.05) and a significant decrease (p < or = 0.01) in malondialdehyde level in the treated group compared to the placebo group. A highly significant decrease (p < or = 0.01) in diuresis, glucose and protein losses through urine were identified in the treated group compared to the placebo group. The superoxide dismutase catalytic activity, urea and cholesterol levels involved nonsignificant changes. The results of biochemical examination show a protective antioxidative and antidiabetic effect of pomiferin. The results of histopathological examination correlate with them only partially.

Daphne decline: what is the causal agent(s)?

A double infection of Daphne mosaic virus (DapMV) and an associated bacilliform virus was observed in the samples of diseased Daphne mezereum shrubs that showed mosaic patterns, precocious leaves reddening, defoliation, repeated flowering with subsequent declining. Extensive aggregations of bacilliform particles (166-370 x 65 nm and 169-233 x 68-78 nm) occurred in the nucleus or perinuclear space of root and leaf tissues suggesting that the virus might belong to the genus Nucleorhabdovirus, family Rhabdoviridae. However, the single bacilliform virions appeared occasionally in the cytoplasm of the leaf cells. We supposed that occurrence of the mixed virus infection could be the cause of D. mezereum decline in the Czech Republic.

[Morin in the therapy of the ischemia-reperfusion damage model of the rat kidney]. / Morin v terapii ischemicko-reperfuzního poskození ledvinné tkáne laboratorního potkana.

The aim of this study was to analyze the protective effects of morin administered during the therapy of reperfusion injury of the laboratory rat kidney. Animals were randomly divided into five groups (n= 10). One group was left intact. Three medicated groups and one placebo group were subjected to ischemia (60 min) and reperfusion of the left kidney. Morin was suspended in a 2 ml of 0.5% Avicel solution and administered orally by a gastric probe at doses of 5, 10, and 20 mg.kg(-1) once a day for 15 days. The placebo group was given only 2 ml of 0.5% Avicel in the same way. On the 15th day, all the animals were exsanguinated and the reperfused kidneys were recovered. Selected biochemical markers in blood were assessed: superoxide dismutase, glutathion peroxidase, total antioxidative capacity, malondialdehyde, creatinine, urea, and uric acid. Creatinine, urea, and total protein were analyzed in urine, and a 24-hour diuresis was recorded. The kidney tissue samples were used for histopathological examination. Morin supported the organism's own defensive reactions against free radicals and decreased lipid peroxidation in the cell membranes and contributed to the recovery of kidney functions. The histopathological results confirm 20 mg x kg(-1) as the most effective dose.

Daphne mosaic virus (DapMV), a new potyvirus from Daphne mezereum in the Czech Republic.

Daphne shrubs with light green rings and mosaic on leaves contained flexuous filamentous virions (696 x 13 nm) and cylindrical inclusions typical of the subdivision III of Edwardson's classification for inclusions induced by members of the family Potyviridae. Decoration tests using antisera to 67 potyviruses revealed distant serological relations among chilli veinal mottle virus, Colombian datura virus, papaya ringspot virus, tobacco vein mottling virus and yam mosaic virus. The 3' terminal region of the virus genome was amplified by RT-PCR using primers specific for cloned and sequenced members of the family Potyviridae. The most similar sequences in the GenBank were those of isolates of wild potato mosaic virus (WPMV) and yam mild mosaic virus (YMMV), originating from Peru and Guadeloupe, respectively. The new sequence had 63.2% and 61.9% nucleotide identity to WPMV and YMMV in the coat protein gene. The results suggest that the Czech isolate from daphne should be regarded as a new member of the genus Potyvirus. The name daphne mosaic virus (DapMV) is suggested for this virus.

Association of phytoplasmas and viruses with malformed clovers.

Plants of Trifolium spp. exhibiting two different kinds of symptoms--phyllody associated with yellowing/reddening, and dwarf growth habit without floral abnormalities--were observed in several areas of the Czechia. Nested polymerase chain reaction (PCR) with phytoplasma specific primers, and restriction fragment length polymorphism (RFLP) analyses of 16SrDNA revealed that phyllody of T. repens was associated with phytoplasmas belonging to the 16SrI-C subgroup. Similar symptoms in T. hybridum and T. pratense plants revealed the presence of phytoplasmas belonging to two subgroups: 16SrI-C and 16SrIII-B. Dwarf disease of cultivated T. pratense plants was associated with more than one agent: 11 of 20 plants examined by PCR/RFLP analysis revealed the presence of phytoplasmas belonging to four distinct subgroups: 16SrI-B, 16SrI-C, 16SrIII-B and 16SrX-A. Moreover, two kinds of bacilliform virions were observed in ultrathin sections of 15 T. pratense plants. Particles occurred mostly in the parenchymatous cells of vascular bundles and were located in the cytoplasm as aggregates within an extended network of membranous cisternae. Phytoplasmas and rhabdoviruses occurred singly, and both together or in co-presence with filamentous virus-like particles.

Occurrence of graft-transmissible virus diseases of the strawberry in the Czech Republic.

Strawberry virus diseases were monitored using a leaf graft bioassay in 17 cultivars of strawberry Fragaria ananassa Duchesne in the Czech Republic. Fragaria vesca indicator clones revealed after grafting several symptoms of strawberry mottle, crinkle, vein banding, and mild yellow edge as well as of mixed virus infections. Isometric virus-like particles (VLPs) ranging from 21 to 50 nm in diameter and flexuous filamentous VLPs (12 by 600-1400 nm) were observed by electron microscopy in negatively stained crude sap preparations. Our results confirmed the complexity of virus diseases of the strawberry and represent the first report on the detection of strawberry mild yellow edge disease (SMYED) and of filamentous particles in this crop in the Czech Republic.

Electron microscopy of two viruses of deadly nightshade (Atropa belladonna L.).

Deadly nightshade plants showing severe necrotic lesions on leaves were observed in southern Bohemia. In negatively stained preparations of spontaneously infected deadly nightshade, artificially inoculated host plants and purified preparations two types of virus particles, isometric ones of about 26 nm in diameter and flexuous ones with length of 765 nm were seen by electron microscopy. The virus with isometric particles was identified as belladonna mottle virus (BMV), indistinguishable serologically from the Hungarian isolate of this virus. Identification of the virus with flexuous particles is discussed. Observations of the ultrastructure revealed the presence of filamentous virus particle aggregates and chloroplasts with peripheral vesicles bounded by double membranes, a feature typical for tymoviruses.

Strawberry vein banding virus--definitive member of the genus Caulimovirus.

The complete DNA sequence (7876 nucleotides) of strawberry vein banding virus (SVBV) has been determined. Seven open reading frames are detected that potentially code for proteins of calculated weight 37.8; 18.3; 16.6; 56.0; 81.1; 59.0 and 12.6 kDa, respectively. Their position on the viral genome is the same as that of the corresponding proteins on the cauliflower mosaic virus (CaMV) genome. Phylogenetic analysis based on the amino acid sequence of this protein shows a closer relationship of SVBV with CaMV, figwort mosaic virus and carnation etched ring virus than with other caulimoviruses.

Occurrence of nepoviruses in Rubus species in the Czech Republic.

The occurrence of arabis mosaic virus (AMV), raspberry ringspot virus (RRV), tomato black ring virus (TBRV), strawberry latent ringspot virus (SLRV) and cherry leaf roll virus (CLRV) in cultivated and wild plants of raspberry and blackberry has been studied in the Czech Republic in 1993-1996. Five hundred and seventy samples were collected at 51 localities and assayed by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). The results represent the first evidence on the occurrence of AMV, RRV, TBRV and SLRV in cultivated Rubus species in the Czech Republic. Isolates AMV M20 and TBRV ML15 which were successfully transmitted by mechanical inoculation and characterized by reactions of differential host plants and by electron microscopy are the first isolates from Rubus from this territory. CLRV was not detected in either cultivated or wild Rubus species.