Determination of protein A I staphylococcus aureus strains isolated from monkeys

Protein A content in Staphylococcus aureus isolated from 6 species of monkeys at the Sukhumi Monkey Nursery has been studied. Protein A has been detected in 73% of the studied strains. One strain isolated from a rhesus macaque has been found to release high amounts of protein A into the environment.     

Genealogical approach to the formation of the winter wheat core collection

The correctness of the genealogical approach to the estimation of genetic diversity has been substantiated. The approach implies a comparison of the estimates of similarity based on genetic markers and coefficients of parentage. A method of the application of the genealogical approach to the formation of the core collection has been developed. The adequacy of the core set has been estimated using Shannon's diversity index adapted for genetic profiles and the correlation between the distributions of ancestors' contributions. A core collection has been formed. This collection contains 25% of the accessions of the original winter wheat (Triticum aestivum L.) collection from the Czech Genebank and represents 70-76% of the intracluster diversity. A decrease in intercluster diversity has not been found in the core collection.     

Determination of amylase activity in bovine serum and its association with the Ami and Amll loci

Amylase activity has been estimated in the sera of 213 cattle by an amyloclastic method with potato amylose as the substrate with the addition of CaCl₂. The method has been developed from that of Street and Close to overcome the problems of the high colour of bovine serum which would otherwise interfere with the assay. A relationship has been found between the serum amylase genotype detected on starch-gel and the total enzyme activity. The enzymes AmI-B and AmII-B in homozygous combinations have been found to have the highest activities. Age of animal has no significant effect on the total enzyme activity. Activity has been investigated in sera from three separate cattle populations and it has been found to be a highly heritable character. Heritability estimates of 49 % and 39 % by sib analysis and 75 % by the twin method have been obtained.     

Genealogical Approach to the Formation of the Winter Wheat Core Collection

The correctness of the genealogical approach to the estimation of genetic diversity has been substantiated. The approach implies a comparison of the estimates of similarity based on genetic markers and coefficients of parentage. A method of the application of the genealogical approach to the formation of the core collection has been developed. The adequacy of the core set has been estimated using Shannon's diversity index adapted for genetic profiles and the correlation between the distributions of ancestors' contributions. A core collection has been formed. This collection contains 25% of the accessions of the entire winter wheat (Triticum aestivum L.) collection from the Czech Genebank and represents 70–76% of the within-cluster diversity. A decrease in between-cluster diversity has not been found in the core collection.     

Primary structure and microheterogeneities of rat chloroleukemia histone H2A (histone ALK, IIbl or F2a2).

Rat chloroleukemia histone H2A, obtained from the F2a2 fraction, has been eluted in two peaks from a Biorex 70 column. The amino acid sequence of rat chloroleukemia histone H2A has been determined and compared to that of calf-thymus histone H2A. The structural studies performed on the tryptic peptides from the maleylated histone and on the thermolysin peptides from the native histone clearly demonstrate the existence of three molecular species of histone H2A depending on the nature of the amino acid residue at positions 16 and 99: H2A-alpha (Ser-16 and Lys-99) accounts for 60% and H2A-betaI (Thr-16 and Arg-99) and H2A-betaII (Ser-16 and Arg-99) for 20% each. A threonine residue at position 16 and a lysine residue at position 99 have been found in calf-thymus histone H2A.     

Improvement in bioreactor productivities using free radicals: HOCl-induced overproduction of xanthan gum from Xanthomonas campestris and its mechanism

Free-radical induction has been employed as a novel strategy to improve bioreactor productivity and, more specifically, the quality and productivity of xanthan gum from Xanthomonas campestris cultures. A 210% increase in xanthan yield and a 20% increase in viscosity (quality) resulted from HOCl (oxidant) treatment. The acetate mass fraction in xanthan gum decreased by 42% and its pyruvate mass fraction increased by 63% as a result of HOCl treatment. The growth rate was almost unaffected by HOCl treatment. A hypothesis to explain the mechanism of xanthan gum overproduction by free-radical induction has been formulated. The significant aspects of the hypothesis, such as SoxS protein binding to the promoter region of the gum gene and the consequent increase in mRNA concentrations, have been experimentally verified.     

Method for removing leukocytes and thrombocytes from preserved blood and packed red cells

A method has been described for removing leukocytes and thrombocytes from preserved ACD blood and from packed red cells of the same type of blood. A system has been used comprising two connected plastic bags model "Fenval". The leukocyte and thrombocyte removal is performed by two consecutive procedures, each comprising centrifugation with subsequent fast sedimentation of erythrocytes. No filters are needed. 97.3 +/- 3.04% leukocyte removal and 96.90 +/- 5.96% thrombocyte removal as regards the preserved blood and 95.65 +/- 8.70% leukocyte removal and 98.44 +/- 0.70% thrombocyte removal as regards the packed red cells have been achieved.     

Purification of extracellular proteinases from B. subtilis SKB 256 by biospecific chromatography

Abstract-A simple and efficient method of preparing highly purified extracellular proteinases of B. subtilis B-1 (SKB 256) has been developed. A sorbent based on sorsilen impregnated with hemoglobin or cytochrome c has been synthesized for this purpose. A significant difference between the efficiency of hemoglobin and cytochrome c as biospecific ligands has been observed: the enzyme yield amounted to 40.6 and 65.6% of the total amount of enzyme adsorbed, respectively. The culture was shown to contain two major proteinase forms with different molecular masses that could be separated by chromatography on a Sephadex G-50 but gave only one band with MW 27 kDa upon denaturing electrophoresis in 12.5% PAG in the presence of 0.1% SDS. The influence of eluent pH, ionic strength and ethanol concentration on the sorption of the proteinases on the biospecific sorbent, as well as on the desorption from it, has been investigated. Positive influence of 20% ethanol on proteinase desorption has been demonstrated.     

Purification, properties and glycoprotein nature of arylsulfatase A from sheep brain.

A simple and rapid method for the purification of arylsulfatase A (EC 3.1.6.1) from sheep brain has been developed. This includes the concanavalin A-Sepharose affinity chromatography and the pH-dependent polymerization and depolymerization of the enzyme. By these methods a homogeneous enzyme was obtained and the enzyme was purified 7180-fold. Sheep brain arylsulfatase A has been shown to be a glycoprotein containing 25% neutral sugar and 0.5% sialic acid. The constituent neutral sugars were identified as glucose and mannose.     

Correlation of x-ray deduced and experimental amino acid sequences of trimethylamine dehydrogenase.

The amino acid sequence of the iron-sulfur-flavoprotein, trimethylamine dehydrogenase, isolated from the bacterium W3A1 has been deduced from the x-ray diffraction pattern obtained at 2.4-A resolution. This sequence has been compared to portions of the primary sequence derived by gas-phase sequencing of isolated peptides obtained from cyanogen bromide and endoprotease Arg-C and Asp-N digestions of the purified enzyme. A consensus sequence has resulted and is comprised of 729 amino acids with Ala at both NH2- and COOH-terminal positions. The consensus sequence contains 13 cysteine residues. Approximately 80% of the sequence has been confirmed by direct sequencing with approximately 81% agreement with the x-ray deduced sequence. The calculated subunit molecular mass of the apoenzyme is 78,899 Da, in good agreement with published values of approximately 83,000. The anomalous scattering map from the native protein has also been shown to provide accurate information about the positions of most of the weak anomalous scattering centers such as sulfur or phosphorus atoms and to complement x-ray or chemical sequencing methods.     

The effect of ultrasound on the antigenic activity of human erythrocytes

The effect of ultrasound (frequency 0.88 MHz, intensity from 0.05 to 1 W/cm2) on alterations in antigenic activity has been investigated in vitro using ABO antigens of human erythrocytes. The existence of threshold doses of ultrasound influence has been found. These doses are shown to be independent of ultrasound intensity. The dependence of the effect on erythrocyte concentration has been established. Individual and group differences in the antigenic resistance to ultrasonic exposure in donors of groups A and B have been revealed. A drop in antigenic activity equal to 97% has been obtained.     

Partial amino acid sequence of a hemocyanin subunit from Palinurus vulgaris

1. Hemocyanin from the spiny lobster Palinurus vulgaris was separated into two fractions, which were designated as subunits a and b. 2. 55% of the amino acid sequence of subunit b has been determined. A comparison with Panulirus interruptus hemocyanins shows 78% sequence identity with subunit a and 56% with subunit c. It has carbohydrate attached to domain one. Two half-cystines have been substituted, indicating that it probably possesses only one disulfide bridge. Heterogeneity has been observed in seven out of 380 positions determined so far. 3. Subunit a is almost identical with subunit b. In contrast to Panulirus interruptus and Panulirus japonicus, Palinurus vulgaris hemocyanin contains no c-type subunit. 4. A position in the tentative evolutionary tree of arthropod hemocyanins based on sequence differences has been assigned to Palinurus vulgaris subunit b.     

Crystallization and preliminary X-ray analysis of L-2-haloacid dehalogenase from Xanthobacter autotrophicus GJ10.

Haloacid dehalogenases are enzymes that cleave carbon-chlorine or carbon-bromine bonds of 2-haloalkanoates. X-ray-quality crystals of L-2-haloacid dehalogenase from the 1,2-dichloroethane-degrading bacterium Xanthobacter autotrophicus GJ10 have been grown at room temperature from 20% PEG 8000, 200 mM sodium formate at pH 6.8-7.0, using macroseeding techniques. The crystals, which diffract in the X-ray beam up to 2.0 A resolution, belong to the spacegroup C2221. Cell parameters are a = 58.8 A, b = 93.1 A, c = 84.2 A. A native data set to 2.3 A has been collected, with a completeness of 97% and an Rsym of 6.0%.     

The infrared and Raman spectra of the duplex of d(GGTATACC) in the crystal show bands due to both the A-form and the B-form of DNA

The deoxyoligonucleotide, d(GGTATACC), forms a duplex structure that crystallizes in the DNA A form. This has been shown by both X-ray diffraction studies and Raman spectroscopy (1,2). The presence of the DNA B form has been reported using diffuse X-ray scattering from a crystal of the closely related sequence d(GGBrUABrUACC)(3). In this paper the infrared spectrum of the d(GGTATACC) crystal is presented and curve resolution of both the Raman and IR spectra have been carried out. The percentage of A and B forms have been estimated. The %B form in the crystal has been estimated from the IR spectra to be about 15% and from Raman to be about 20%. Moreover the IR spectrum of the A conformation in the crystal is slightly different from the IR spectrum of the A conformation in polynucleotide fibers in particular in the region of the phosphate stretching vibrations and of the in-plane double bond vibrations of the bases. We show that it is feasible to obtain IR as well as Raman spectra of small crystals of oligonucleotides and that this is a good method of identifying all of the different conformations that may be in the crystal.     

The Infrared and Raman Spectra of the Duplex of d(GGTATACC) in the Crystal Show Bands Due to Both the A-form and the B-form of DNA

Abstract

The deoxyoligonucleotide, d(GGTATACC), forms a duplex structure that crystallizes in the DNA A form. This has been shown by both X-ray diffraction studies and Raman spectroscopy (1,2). The presence of the DNA B form has been reported using diffuse X-ray scattering from a crystal of the closely related sequence d(GGBrUABrUACC)(3). In this paper the infrared spectrum of the d(GGTATACC) crystal is presented and curve resolution of both the Raman and IR spectra have been carried out. The percentage of A and B forms have been estimated. The %B form in the crystal has been estimated from the IR spectra to be about 15% and from Raman to be about 20%. Moreover the IR spectrum of the A conformation in the crystal is slightly different from the IR spectrum of the A conformation in polynucleotide fibers in particular in the region of the phosphate stretching vibrations and of the in-plane double bond vibrations of the bases. We show that it is feasible to obtain IR as well as Raman spectra of small crystals of oligonucleotides and that this is a good method of identifying all of the different conformations that may be in the crystal.     

Hordein locus polymorphism of cultivated barley (Hordeum vulgare L.) in Turkey

Starch gel electrophoresis has been used to study the polymorphism of hordeins encoded by the Hrd A, Hrd B, and Hrd F loci in 93 landrace specimens of barley assigned to 17 ancient provinces located in modem Turkey. Forty-five alleles of Hrd A with frequencies of 0.11-29.34%, 51 alleles of Hrd B with frequencies of 0.11-8.07%, and 5 alleles of Hrd F with frequencies of 0.75-41.29% have been detected. Cluster analysis of the matrix of allele frequencies has demonstrated that barley populations from different old provinces of Turkey are similar to one another. Cluster structure of local barley populations has been found, most populations (82%) falling into three clusters. The first cluster comprises barley populations from six provinces (Thracia, Bithynia, Pontus, Lydia, Cappadocia, and Armenia); the second cluster, populations from five provinces (Paphlagonia, Galatia, Lycaonia, Cilicia, and Mesopotamia); and the third one, populations from three provinces (Phrygia, Karia, and Lycia). Barley populations from Mysia, Pamphlya, and Syria do not fall in any cluster.     

Characterization of a human chromosome 8 cosmid library constructed from flow-sorted chromosomes.

A cosmid library for human chromosome 8 has been constructed from flow-sorted chromosomes in the vector sCos-1. This library is 85% human and has been arrayed into 210 microtiter plates representing four genome equivalents. Cosmids have been isolated with 10 of 11 probes representing nine different loci from chromosome 8.     

Transglycosylation activity of Dictyoglomus thermophilum amylase A

Amylase A from Dictyoglomus thermophilum is a thermophilic enzyme and has about 40% identity with 4-alpha-glucanotransferase (GTase) from Thermococcus litoralis, and both of these enzymes belong to family 57 glycosyl hydrolase. Since the transglycosylation activity of T. litoralis GTase has been well characterized, the substrate specificity and reaction products of amylase A from D. thermophilum were examined. alpha-1,4 Glucan was produced from maltooligosaccharides, and glucoamylase-resistant molecules (cycloamyloses) were produced from longer chain amylose (average molecular mass 200 kDa). It has been reported that amylase A from D. thermophilum hydrolyzes starch, but in this study it was found that the enzyme was also able to use maltooligosaccharides and long chain amylose as substrate and has transglycosylation activity.