Immunoprotective properties of transgenic plants expressing E2 glycoprotein from CSFV and cysteine protease from Fasciola hepatica.

Immune responses were elicited in laboratory animals after oral vaccination by transgenic plants (lettuce and alfalfa) expressing the E2 glycoprotein of Classical Swine Fever Virus (CSFV) or cysteine protease from Fasciola hepatica. ELISA analyses demonstrated that the oral route is effective in inducing a specific antibody response against these antigens in mice.

The variable part of the dnaK gene as an alternative marker for phylogenetic studies of rhizobia and related alpha Proteobacteria.

DnaK is the 70 kDa chaperone that prevents protein aggregation and supports the refolding of damaged proteins. Due to sequence conservation and its ubiquity this chaperone has been widely used in phylogenetic studies. In this study, we applied the less conserved part that encodes the so-called alpha-subdomain of the substrate-binding domain of DnaK for phylogenetic analysis of rhizobia and related non-symbiotic alpha-Proteobacteria. A single 330 bp DNA fragment was routinely amplified from DNA templates isolated from the species of the genera, Azorhizobium, Bradyrhizobium, Mesorhizobium, Rhizobium and Sinorhizobium, but also from some non-symbiotic alpha Proteobacteria such as Blastochloris, Chelatobacter and Chelatococcus. Phylogenetic analyses revealed high congruence between dnaK sequences and 16S rDNA trees, but they were not identical. In contrast, the partition homogeneity tests revealed that dnaK sequence data could be combined with other housekeeping genes such as recA, atpD or glnA. The dnaK trees exhibited good resolution in the cases of the genera Mesorhizobium, Sinorhizobium and Rhizobium, even better than usually shown by 16S rDNA phylogeny. The dnaK phylogeny supported the close phylogenetic relationship of Rhizobium galegae and Agrobacterium tumefaciens (R. radiobacter) C58, which together formed a separate branch within the fast-growing rhizobia, albeit closer to the genus Sinorhizobium. The Rhizobium and Sinorhizobium genera carried an insertion composed of two amino acids, which additionally supported the phylogenetic affinity of these two genera, as well as their distinctness from the Mesorhizobium genus. Consistently with the phylogeny shown by 16S-23S rDNA intergenic region sequences, the dnaK trees divided the genus Bradyrhizobium into three main lineages, corresponding to B. japonicum, B. elkanii, and photosynthetic Bradyrhizobium strains that infect Aeschynomene plants. Our results suggest that the 330 bp dnaK sequences could be used as an additional taxonomic marker for rhizobia and related species (alternatively to the 16S rRNA gene phylogeny).

Low sequence similarity and gene content of symbiotic clusters of Bradyrhizobium sp. WM9 (Lupinus) indicate early divergence of "lupin" lineage in the genus Bradyrhizobium.

Two sequenced nodulation regions of lupin Bradyrhizobium sp. WM9 carried the majority of genes involved in the Nod factor production. The nod region I harbored: nolA, nodD, nodA, nodB, nodC, nodS, nodI, nodJ, nolO, nodZ, fixR, nifA, fixA, nodM, nolK and noeL. This gene arrangement resembled that found in the nodulation region of Bradyrhizobium japonicum USDA110, however strain WM9 harbored only one nodD gene copy, while the nodM, nolK and noeL genes had no counterparts in the 410 kb symbiotic region of strain USDA110. Region II harbored nolL and nodW, but lacked an nodV gene. Both regions carried ORFs that lacked similarity to the published USDA110 sequences, though they had homologues in symbiotic regions of Rhizobium etli, Sinorhizobium sp. NGR234 and Mesorhizobium loti. These differences in gene content, as well as a low average sequence identity (70%) of symbiotic genes with respect to B. japonicum USDA110 were in contrast with the phylogenetic relationship of USDA110 and WM9 revealed by the analysis of 16S rDNA and dnaK sequences. This most likely reflected an early divergence of symbiotic loci, and possible co-speciation with distinct legumes. During this process the loss of a noel gene and the acquisition of a nolL gene could be regarded as an adaptation towards these legumes that responded to Nod factors carrying 4-O-acetylfucose rather than 2-O-methylfucose. This explained various responses of lupins and serradella plants to infection by mutants in nodZ and nolL genes, knowing that serradella is a stringent legume while lupins are more promiscuous legumes.

[Metabolic control in young children with type 1 diabetes treated with continuous subcutaneous insulin infusion (insulin pump)]. / Wyrównanie metaboliczne u malych dzieci z cukrzyca typu 1 leczonych metoda ciaglego podskórnego wlewu insuliny (pompa insulinowa).

UNLABELLED: Intensive insulin therapy is a method of choice in the management of patients with type 1 diabetes. Its administration in the youngest children is limited by little or no acceptance of multiple injections and a typical fear of needles and syringes. In recent years more and more frequently the method of multiple daily injections (MDI) of insulin is being replaced by the method of continuous subcutaneous insulin infusion (CSII) even in the youngest children. OBJECTIVE: Evaluation of the safety and efficacy of CSII method in children at prepubertal age. MATERIAL AND METHODS: There were 61 children under 10 years of age with type 1 diabetes recruited for the study (33 boys, 28 girls). CSII method was implemented for the period of minimum 6 months. In the group of 21 children CSII method was the first method of their therapy (it was administered at the time of diagnosis). Mean duration of diabetes was 3.0 years +/-1.87 year, mean age at diagnosis was 3.82+/-2.19 years and mean duration of CSII treatment was 1.4 +/-0.75 year. The average HbA1c at the baseline for all children was 8.7+/-1.4%. RESULTS: In the group where CSII therapy was implemented as the first method of management, mean duration of treatment was 1.5 years, mean HbA1c decreased after first 3 months from 9.6+/-1.68% to 7.22+/-0.99% (p<0.05). After 12 and 24 months the value further decreased to 7.01+/-0.57%. In the group that was earlier treated with MDI method (n=40), mean value of HbA1c decreased after 3 months from 8.27+/-1.4% to 7.6 +/-0.86% (p<0.05), after 12 months it further decreased to 7.37+/-0.86%, after 24 months its mean value was 7.53%. The number of patients with HbA1c >8% decreased from 58.4% to 10%. Adverse events were observed only in the group that was earlier treated with the MDI method. There were 3 incidences of severe hypoglycaemia, 2 incidences of diabetic ketoacidosis, 2 incidences of infection at the needle site (in one case the surgical attention was necessary). After two years of the trial there was a statistically significant difference in the mean value of HbA1c between children that used CSII method from the moment of their diagnosis (HbA1c=7.01%) and those who were earlier treated with MDI method (HbA1c=7.53 +/-0.73%). In both groups the daily insulin requirement was similar (CSII method 0.69+/-0.2 unit/kg/day, MDI method 0.75+/-0.19 unit/kg/day). CONCLUSIONS: The method of continuous subcutaneous insulin infusion (CSII) provides good and sustained metabolic control in the youngest children with type 1 diabetes. Administering of that method from the very beginning of the diabetes treatment may decrease the risk of acute complications.