Relationship between soluble carbohydrate level and tolerance of meadow fescue callus to Bipolaris sorokiniana (Sacc.) Shoem. and Drechslera dictyoides (Drechsl.) Shoem. metabolites

The aim of the presented work was the search for the relationship between the level of soluble carbohydrates in callus tissues of eight meadow fescue (Festuca pratensis Huds.) cultivars and their growth ability on media containing Bipolaris sorokiniana and Drechslera dictyoides metabolites. Calli were induced from mature grains using the method previously described (Płażek 1994). Callus obtained from single caryopsis was cut into three pieces which were weighted and put on the media with or without pathogen metabolites. Tissue selection was performed by means of “double-layer culture” technique (Lepoivre et al. 1986). After two-week culture in the darkness at temp. of 25°C the calli were weighted again. The sugar level in tissue was measured by means of colorimetric method of Klein & Weissman according to Snell (1961). Fresh mass decrease of calli developing on the media with fungus metabolites was observed by all studied object. The tolerance of calli of the tested cultivars to metabolites of both pathogens was significantly different. However, significant similarity between the tolerance of calli of particular varietes to both fungi was noted. The soluble carbohydrate contents in control tissue of all studied cultivars were similar and their values ranged between 2.4 and 3 % of fresh mass. B. sorokiniana metabolites caused a significant decrease of the sugar content in calli, while D. dictyoides metabolites did not decrease the sugar level.     

Activities of ligninolytic enzymes in some white-rot basidiomycete strains after recovering from cryopreservation in liquid nitrogen

Fourteen strains of white-rot basidiomycetes belonging to eight species of two genera (Inonotus and Pholiota) were tested for their ability to maintain the production of laccase, peroxidase and manganese-dependent peroxidase (enzymes involved in lignin biodegradation) after a short-time preservation in liquid nitrogen with different cryoprotectives (glycerol, dimethyl sulfoxide). No negative effect of cryopreservation or the used cryoprotective on production of the ligninolytic enzymes was found in the fungi tested.     

High-performance thin-layer chromatographic separation of ranitidine hydrochloride and two related compounds

Ranitidine hydrochloride and its two related compounds, used in the USP TLC purity testing of the drug, were separated on a high-performance thin-layer chromatography (HPTLC) RP-18 WF_254S precoated plate using methanol–3% NH₄OH (4:1, v/v) as the mobile phase. The main advantage of the proposed HPTLC system over the USP TLC system for testing the purity of ranitidine is a better and more efficient separation of these three compounds in a shorter time and with less consumption of solvents. The system is promising from the point of view of the development of a new method for the TLC purity testing of ranitidine hydrochloride. A video system was used for imaging thin-layer chromatograms. Direct UV densitometric quantitation of the three compounds and a model for the calculation of analytical performance parameters is presented in the second part of the paper.     

The Human Cystathionine β-Synthase (CBS) Gene: Complete Sequence, Alternative Splicing, and Polymorphisms

Cystathionine β-synthase [CBS; -serine hydro-lyase (adding homocysteine), EC 4.2.1.22] catalyzes the first committed step of transsulfuration and is the enzyme deficient in classical homocystinuria. In this report, we describe the molecular cloning and the complete nucleotide sequence of the human CBS gene. We report a total of 28,046 nucleotides of sequence, which, in addition to the CBS gene, contains 5 kb of the 5′ flanking region. The human CBS gene contains 23 exons ranging from 42 to 209 bp. The 5′ UTR is formed by 1 of 5 alternatively used exons and 1 invariably present exon, while the 3′ UTR is encoded by exons 16 and 17. We also describe the identification of two alternatively used promoter regions that are GC rich (80%) and contain numerous putative binding sites for Sp1, Ap1, Ap2, and c-myb, but lack the classical TATA box. The CBS locus contains an unusually high number ofAlurepeats, which may predispose this gene to deleterious rearrangements. Additionally, we report on a number of DNA sequence repeats that are polymorphic in North American and European Caucasians.     

Ammonium uptake in alpine streams in the High Tatra Mountains (Slovakia)

Uptake of NH _inf4 ^sup+ -N by streambed biota of mountain brooks was studied in the alpine zone of the High Tatra Mountains. Experiments were performed involving in situ dosing of ammonium directly to the acidified stream and incubations of ammonium and streambed bryophytes in enclosures within a range of pH from 4.45 to 8.10.NH _inf4 ^sup+ -N uptake length decreased with decreasing stream discharge, while comparable values of discharge-normalized uptake lengths were found during two in situ experiments.Maximum uptake rates of NH _inf4 ^sup+ -N obtained during the incubation of bryophytes (6 to 11 mg m^–2 h^–1) were comparable with results of two in situ experiments (8 and 12 mg m^–2 h^–1). The average NH _inf4 ^sup+ -N uptake rates observed during incubations lasting 3 to 5 hours (4.3 mg m^–2 h^–1) were not related to the pH of stream water. Nitrification of about 50% of the NH _inf4 ^sup+ -N added was observed in non-acidified streams, but was negligible in acidified streams. Significant photoinhibition of nitrification was observed in non-acidified streams during enclosure experiments.     

Chlorophyll-phosphorus relationship in acidified lakes of the High Tatra Mountains (Slovakia)

Concentrations of total phosphorus (TP) and chlorophyll a (CHLA) were measured in 28 lakes in the High Tatra Mountains (Slovakia) from 1983 to 1990. The relationship between log CHLA and log TP in the Tatra lakes is similar to relationships developed for lakes in other regions, but variation is higher. A part of this variation is caused by acidification of the lakes. In the lakes with pH between 4.9 and 6.3 the CHLA concentrations are often extremely low while TP concentrations decreased, but not as drastically.     

PSTV infection in tobacco (Nicotiana tabacum L.) transformed with PSTV cDNA

Tobacco cv. White Burley was transformed with disarmed expression vector pCB1314 containing dimeric cDNA of potato spindle tuber viroid (PSTV, severe strain) in plus orientation regulated from the mannopine promoter. Amount of PSTV specific (?) and (+) sequences and PSTV circular forms was measured in transformed tobacco stock and compared with PSTV content in untransformed tomato and tobacco grafts. It follows from the results that lower rate of accumulation of PSTV in tobacco as compared with tomato is due to less intensive viroid transportation through the cytoplasm and/or cell to cell moverment, whereas both, viroid replication and processing showed comparable characteristics in tomato and tobacco with respect to accumulation of minus and plus strands and circular forms in infected tissues. Despite of accumulation of viroid in comparable amount in both transformed tobacco and infected tomato, no expression of any morphological symptom of disease was observed in transgenic tobacco.     

Production of avermectin A2a and monoglycosides A2a and B2a by a strain ofStreptomyces avermitilis

A strain ofStreptomyces avermitilis producing almost predominantly avermectin A2a and monoglycosides A2a and B2a was described. Methods of analytical and preparative high performance liquid chromatography were used for comparison of chromatographic profiles and product isolation, respectively. Identification of isolated compounds was based on¹³C-NMR spectrometric results.     

Palatoquadrate in a Devonian fish Eusthenopteron Evidence of its dual origin

The palatoquadrate in Eusthenopteron displays certain variation concerning the extent of its commissural lamina, and the presence of imprints of vessels and nerves on the outer surface of this middle part of the element. Comparison with some contemporary fishes revealed that branches of the arteria ophthalmica magna or of the ramus palatinus posterior VII could have produced these imprints. The grooves are connected with canals piercing the bone along its pars pterygoquadrata. Possible explanation is that the autopalatine and pterygoquadrate portions of the palatoquadrate were free in early developmental stages and later become interconnected by cartilage and bone. The dual nature of the palatoquadrate is shown to occur not only in the development of some Recent fishes, but in extinct groups (e. g., placoderms) as well.     

Evolution of parallel β/α-barrel enzyme family lightened by structural data on starch-processing enzymes

The parallel /-barrel domain consisting of eight parallel -sheets surrounded by eight -helices has been currently identified in crystal structures of more than 20 enzymes. This type of protein folding motif makes it possible to catalyze various biochemical reactions on a variety of substrates (i.e., it seems to be robust enough so that different enzymatic functionalities could be designed on it). In spite of many efforts aimed at elucidation of evolutionary history of the present-day /-barrels, a challenging question remains unanswered: How has the parallel /-barrel fold arisen? Although the complete sequence comparison of all /-barrel amino acid sequences is not yet available, several sequence similarities have been revealed by using the highly conserved regions of -amylase as structural templates. Since many starch-processing enzymes adopt the parallel /-barrel structure these enzymes might be useful in the search for evolutionary relationships of the whole parallel eight-folded /-barrel enzyme family.