A Case Report of Porphyria Cutanea Tarda with Hepatitis-C Virus Co-infection.

Porphyria cutanea tarda is a rare disorder of heme metabolism due to deficiency of the enzyme uroporphyrinogen decarboxylase which is manifested as some typical dermatological features and hepatic dysfunction. The Hepatitis-C virus co-infection is common and it can be aggravated by other environmental factors. We report a case of porphyria cutanea tarda in a 37-year-old woman, who presented with recurrent skin blisters and has concomitant Hepatitis-C virus infection. She was taking oestrogen containing oral contraceptive pill for a long duration. The diagnosis of porphyria cutanea tarda was considered on the basis of clinical features and high level of urine porphyrin level. She was put on hydroxychloroquine and combination drugs for Hepatitis-C virus with significant improvement after 3 months of therapy.

Raman Staircase in Charge Transfer SERS at the Junction of Fusing Nanospheres.

We present a theoretical analysis of Raman intensities for a molecule that bridges a current carrying junction. Experimental data is used to estimate parameters for the theoretical model. The recently reported staircase of Raman intensities observed during the fusion of nanodumbbell is reproduced.

Construction of a BAC library and a physical map of a major QTL for CBB resistance of common bean (Phaseolus vulgaris L.).

A major quantitative trait loci (QTL) conditioning common bacterial blight (CBB) resistance in common bean (Phaseolus vulgaris L.) lines HR45 and HR67 was derived from XAN159, a resistant line obtained from an interspecific cross between common bean lines and the tepary bean (P. acutifolius L.) line PI319443. This source of CBB resistance is widely used in bean breeding. Several other CBB resistance QTL have been identified but none of them have been physically mapped. Four molecular markers tightly linked to this QTL have been identified suitable for marker assisted selection and physical mapping of the resistance gene. A bacterial artificial chromosome (BAC) library was constructed from high molecular weight DNA of HR45 and is composed of 33,024 clones. The size of individual BAC clone inserts ranges from 30 kb to 280 kb with an average size of 107 kb. The library is estimated to represent approximately sixfold genome coverage. The BAC library was screened as BAC pools using four PCR-based molecular markers. Two to seven BAC clones were identified by each marker. Two clones were found to have both markers PV-tttc001 and STS183. One preliminary contig was assembled based on DNA finger printing of those positive BAC clones. The minimum tiling path of the contig contains 6 BAC clones spanning an estimated size of 750 kb covering the QTL region.

End of life.

Dying, but not the death, is an essential problem. The more we believe that death ends everything, the more we fear from death. A human spirit only hardly copes with this fact. All religions want to cut this fear. They highlight that present life continues and human spirit lives further on, in another postmortem dimension. Authors evaluated death of 142 patients, among which 45 (32%) died at home, 74 (52%) in hospital, 34 (24%) died among family relatives and 56 (39%) without the presence of relatives. Most of the dying patients wish to stay with their family or relatives at the end of life (end of life decision). If this wish cannot be fulfilled, then a palliative care seems to be the most suitable alternative for an individual in terminal stage in modern society. In the Presov region, there is a lack of hospices and palliative care does not cover the needs of terminally ill patients (Tab. 6, Ref: 41).

The cloning and characterization of alpha-galactosidase present during and following germination of tomato (Lycopersicon esculentum Mill.) seed.

alpha-Galactosidase (EC 3.2.1.22) is present in the embryo, micropylar and lateral endosperm of seeds of tomato during and following germination. Its activity is unchanged even when germination of the seeds is prevented by an osmoticum. It is also present in the developing and mature dry seed. A cDNA clone for tomato seed alpha-galactosidase (LeaGal) has been isolated and the characteristics of the protein deduced; the predicted molecular mass of the mature enzyme is 39.8 kDa, with a pI of 4.91. The tomato alpha-galactosidase has a high homology (>62%) at the amino acid level with that of other plant alpha-galactosidases. A hydrophobic signal peptide region is identified which is indicative that the enzyme enters the lumen of the endoplasmic reticulum during its translation, prior to its export to the protein body or cell wall, the presumed sites of its substrates. Using amino acid alignment and phylogenetic analysis, key amino acids have been identified, and relationships to other alpha-galactosidases inferred. Southern hybridization analyses show that the enzyme is derived from a single gene (for which a partial sequence has been obtained) and yet there are at least three different isoforms within the seed; post-translational modifications are thus presumed to occur. From Northern hybridization studies it is evident that alpha-galactosidase transcripts are present in the lateral and micropylar endosperm during and following germination, and also to a lesser extent in the embryo.

Endo-beta-mannanase is present in an inactive form in ripening tomato fruits of the cultivar Walter.

Fruits of the tomato cultivar Walter undergo normal development to the red-ripe stage but, unlike those of the cultivar Trust, they do not produce any active endo-beta-mannanase. Reasons for this failure to produce the enzyme were sought. The cv. Walter contains genes for endo-beta-mannanase, as shown by Southern blot analysis, and transcripts for the enzyme are present in ripening fruits, as revealed using Northern hybridization. Moreover, the enzyme protein is detectable by Western blots using an endo-beta-mannanase-specific antibody from tomato. In addition, the inactive enzyme is localized appropriately in the wall regions of the outer layers of the fruit (skin and outer pericarp). Mixing inactive fruit extracts of cv. Walter, in excess, with extracts from cv. Trust fruits, which contain active enzyme, leads to an increase rather than a reduction in enzyme activity, showing that there are no inhibitors of endo-beta-mannanase in cv. Walter fruits. Similar results were obtained with fruits of the tomato cv. Heinz 1439. In contrast to the situation in fruits, the seeds of both cvs Walter and Heinz 1439 produce active enzyme, especially following germination.

Endo-beta-mannanase activity increases in the skin and outer pericarp of tomato fruits during ripening.

Activity of endo-beta-mannanase increases during ripening of tomato (Lycopersicon esculentum Mill.) fruit of the cultivar Trust. beta-Mannoside mannohydrolase is also present during ripening, but its pattern of activity is different from that of endo-beta-mannanase. The increase in endo-beta-mannanase activity is greatest in the skin, and less in the outer and inner pericarp regions. This enzyme is probably bound to the walls of the outermost cell layers of the fruit during ripening, and it requires a high-salt buffer for effective extraction. The enzyme protein, as detected immunologically on Western blots, is present during the early stages of ripening, before any enzyme activity is detectable. The mRNA for the enzyme is also present at these stages; endo-beta-mannanase may be produced and sequestered in a mature-sized inactive form during early ripening. Most non-ripening mutants of tomato exhibit reduced softening and lower endo-beta-mannanase activity, but a cause-and-effect relationship between the enzyme and ripening is unlikely because some cultivars which ripen normally do not exhibit any endo-beta-mannanase activity in the fruit.

Polysaccharide hydrolases in germinated barley and their role in the depolymerization of plant and fungal cell walls.

Cell wall degradation is an important event during endosperm mobilization in the germinated barley grain. A battery of polysaccharide and oligosaccharide hydrolases is required for the complete depolymerization of the arabinoxylans and (1 --> 3,1 --> 4)-beta-glucans which comprise in excess of 90% by weight of these walls. The (1 --> 3,1 --> 4)-beta-glucan endohydrolases release oligosaccharides from their substrate and are probably of central importance for the initial solubilization of the (1 --> 3,1 --> 4)-beta-glucans, but beta-glucan exohydrolases and beta-glucosidases may be important additional enzymes for the conversion of released oligosaccharides to glucose. The latter enzymes have recently been purified from germinated barley and characterized. There is an increasing body of evidence to support the notion that the (1 --> 3,1 --> 4)-beta-glucan endohydrolases from germinated barley evolved from the pathogenesis-related (1 --> 3)-beta-glucanases which are widely distributed in plants and which hydrolyse polysaccharides that are abundant in fungal cell walls. Arabinoxylan depolymerization is also mediated by a family of enzymes, but these are less well characterized. (1 --> 4)-beta-Xylan endohydrolases have been purified and the corresponding cDNAs and genes isolated. While the presence of (1 --> 4)-beta-xylan exohydrolases and alpha-L-arabinofuranosidases has been reported many times, the enzymes have not yet been studied in detail. Here, recent advances in the enzymology and physiology of cell wall degradation in the germinated barley grain are briefly reviewed.

Structure, hormonal regulation, and chromosomal location of genes encoding barley (1-->4)-beta-xylan endohydrolases.

A gene encoding (1-->4)-beta-xylan xylanohydrolase (EC 3.2.1.8) isoenzyme X-I has been isolated from a barley genomic library and the nucleotide sequence of a 2704-bp fragment defined. The gene contains a single intron of 91 bp in the coding region of the mature enzyme and additional introns may be present in the 5'-untranslated region. Expression of the xylanase gene is restricted to the aleurone layer of germinated grain, where the phytohormone gibberellic acid induces both transcriptional activity of the gene and the secretion of active enzyme from the layers. Abscisic acid abolishes the gibberellic acid induction of xylanase gene expression. The hormonal responses are consistent with the presence of promoter sequences, all of which are within 150 bp of the putative transcription start site, that have been implicated as cis-acting elements within gibberellic acid response complexes in plant genes. The elements include a pyrimidine box, CTCTTTCC, together with TAACGAC and TATCCAT boxes. Three genes encode (1-->4)-beta-xylanase isoenzymes in barley and these have been mapped on the barley genome using two doubled haploid populations and seven wheat-barley addition lines. The three xylanase genes are closely linked on the long arm of chromosome 7 (5H). No recombination was detected between the genes in 234 doubled haploid lines. The genes are flanked by the RFLP markers CDO506 on the proximal side and PSR370 at the distal end.

Molecular cloning of cDNAs encoding (1-->4)-beta-xylan endohydrolases from the aleurone layer of germinated barley (Hordeum vulgare).

Heteroxylans are major constituents of cell walls in the graminaceous monocotyledons. Degradation of walls in the starchy endosperm of germinated cereal grains is mediated, in part at least, by the action of (1-->4)-beta-xylan endohydrolases (EC 3.2.1.8). Complementary DNAs encoding (1-->4)-beta-xylan endohydrolases from the aleurone layer of germinated barley have been isolated and characterized. Southern blot analyses suggest that the enzymes are derived from a family of 3 or 4 genes, and cDNAs corresponding to two of these genes have been sequenced. The amino acid sequence deduced from one cDNA almost exactly matches the amino acid sequence determined previously from the purified enzyme. This enzyme is designated (1-->4)-beta-xylan endohydrolase isoenzyme X-I. The mature enzyme consists of 395 amino acid residues, has a calculated M(r) of ca. 44600 and an isoelectric point of 6.1, and is likely to adopt an (alpha/beta)8 barrel conformation. The amino acid sequence of the barley (1-->4)-beta-xylan endohydrolase encoded by the other cDNA, which is designated isoenzyme X-II, shows ca. 13% sequence divergence compared with isoenzyme X-I. Both enzymes exhibit sequence and structural similarities with microbial xylanases. Expression of the genes in germinated grain appears to be confined largely to the aleurone layer, and no mRNA transcripts could be detected in young vegetative tissues.