Recombinant human interleukin-1 beta: new possibilities for the prophylaxis and correction of toxic myelodepression in patients with malignant tumors. II. Phase II study of the protective effect of recombinant human interleukin-1 beta on myelodepression induced by chemotherapy in cancer patients.

The clinical study of human recombinant IL-1beta as a leukopoiesis protector, administered intravenously and subcutaneously, simultaneously with the chemotherapy, confirmed the "protective" effect of this cytokine in patients with grade II leukopenia induced by previous cycles of chemotherapy. The highly protective effect of IL-1beta was proved by the normalization of absolute peripheral blood granulocyte counts over 5 days, from the start of hemostimulation in patients with leukopenia induced by previous cycles of chemotherapy, despite the continuation of cytostatic treatment. Changing the method of IL-1beta administration from the intravenous to the subcutaneous route had no statistically significant effect on the protective influence of IL-1beta. The time taken for complete restoration of leukopoiesis was almost the same for both methods, 5 +/- 1 days and 7 +/- 1 days for s.c. and i.v. methods respectively. This effect of IL-1beta, not seen in other hematopoietic growth factors such as GM-CSF and G-CSF, allows greater flexibility as regards chemotherapy and provides increased effectiveness of anticancer treatments.     

The pleiotropic function of the phosphoenolpyruvate-dependent phosphotransferase system in bacteria. Communication II

The structure and function of regulators and anti-terminators is under discussion in gram-positive bacteria. The regulators of lichen and levan operons (LiR and LevR) as well as the implementation of both gram-positive and negative regulations of operons by them are in the focus of attention. Po-independent termination is regarded by the example of the regulatory activity for the utilization systems of glucose (GlcT) beta-glucosides (LicT), sucrose (low-efficiency system SacY-SacX) and of glycerin (GlcP). Changes in the functional activity of the above systems, which are dependent on a condition of anti-terminators (phosphorylated or dephosphorylated forms and an ability to demirelize etc.) are regarded from the viewpoint of a possibility of occurrence of catabolic repression.     

Nitrogen assimilation enzymes in Bacillus subtilis mutants with hyperproduction of riboflavin

Three groups of the nitrogen assimilation cycle enzymes (glutamate synthases (GTS), glutamine synthases (GS), and glutamate dehydrogenases (GD)) were studied in Bacillus subtilis strains with hyperproduction of riboflavin (vitamin B2). It was found that in all strains tested activity of GS was virtually the same, activity of GD was absent, and activity of GTS was reduced. In strains 41 and 24, riboflavin producers, activity of GTS was 30-60% the enzyme activity in the original strain (wild-type RosR). The most pronounced decrease in the activity of GTS (0-12% relative to RosR) was observed in the strain AS5, which had the highest level of biosynthetic activity relative to the other strains. According to the results of determination of the sensitivity of induction of beta-xylosidase to glucose- and fructose-induced catabolic repression, none of the strains studied was characterized by disorders in the protein CcpA, a global regulator of the catabolic repression in gram-positive bacteria, which is required for reducing amination and resulting activation of biosynthesis of glutamic acid in cell. It was suggested that mutations responsible for partial or complete inhibition of GTS biosynthesis caused an increase in the intracellular pool of glutamine. The intracellular pool of glutamine is a nitrogen source for riboflavin in cell. It follows from the results of this work that there is a trend toward an increase in the rate of biosynthesis of vitamin B2 in mutants with inhibited GTS activity. However, the complexity of the processes of regulation of nitrogen assimilation enzymes makes it difficult to find a distinct correlation between GTS activity and riboflavin biosynthesis in these strains.     

Cloning of the genes of hemolytic factors of Bordetella pertussis in Escherichia coli

The gene library of B. pertussis strain No. 475 (serovar 1. 2. 3.) was obtained on plasmid pUC19 in E. coli strain JM107. The average size of the cloned fragments of chromosomal DNA was 4.9 Kb. Out of 1,700 tested recombinant clones, six caused visible hemolysis on petri dishes with agar containing 4% of sheep red blood cells after 16-hour incubation at 37 degrees C. Only two out of four isolated plasmids were similar in Pst I restriction. Consequently, the existence of several different genes responsible for the hemolytic factors of B. pertussis may be assumed. None of the cloned fragments had Hind III sites. Clones harboring hybrid plasmids possessed different modes of hemolysis, which may be indicative of gene expression in E. coli.     

An essential saccharide binding domain for the mAb 2C7 established for Neisseria gonorrhoeae LOS by ES-MS and MSn

A study of bacterial surface oligosaccharides were investigated among different strains of Neisseria gonorrhoeae to correlate structural features essential for binding to the MAb 2C7. This epitope is widely expressed and conserved in gonococcal isolates, characteristics essential to an effective candidate vaccine antigen. Sample lipooligosaccharides (LOS), was prepared by a modification of the hot phenol-water method from which de-O-acetylated LOS and oligosaccharide (OS) components were analyzed by ES-MS-CID-MS and ES-MSnin a triple quadrupole and an ion trap mass spectrometer, respectively. Previously documented natural heterogeneity was apparent from both LOS and OS preparations which was admixed with fragments induced by hydrazine and mild acid treatment. Natural heterogeneity was limited to phosphorylation and antenni extensions to the alpha-chain. Mild acid hydrolysis to release OS also hydrolyzed the beta(1-->6) glycosidic linkage of lipid A. OS structures were determined by collisional and resonance excitation combined with MS and multistep MSn which provided sequence information from both neutral loss, and nonreducing terminal fragments. A comparison of OS structures, with earlier knowledge of MAb binding, enzyme treatment, and partial acid hydrolysis indicates a generic overlapping domain for 2C7 binding. Reoccurring structural features include a Hepalpha(1-->3)Hepbeta(1-->5)KDO trisaccharide core branched on the nonreducing terminus (Hep-2) with an alpha(1-->2) linked GlcNAc (gamma-chain), and an alpha-linked lactose (beta-chain) residue. From the central heptose (Hep-1), a beta(1-->4) linked lactose (alpha-chain), moiety is required although extensions to this residue appear unnecessary.      

Cloning and the expression of the hemolysin gene of Leptospira pomona pomona in Escherichia coli

The library of Leptospira pomona genes was obtained on phage vector AL 47.1. From this library a recombinant phage carrying the hemolysin gene was selected. The DNA fragment (7.7 kb) of this phage containing the hemolysin gene was subcloned on plasmid pUC19. E. coli clones with hybrid plasmid pDR7 were shown to be hemolytic, but the secretion of hemolysin by E. coli into the culture medium was not observed.     

Isolation and properties of merodiploid strains with F-factor including the pts-region of the Escherichia coli K-12 chromosome]

A technique of hybridization of haploid methanol-utilizing yeast Pichia pinus MH4 is worked out using UV- and N-nitrosoguanidine-induced auxotrophic mutants. Vegetative diploid cultures are isolated. Tetrad analysis and random spore analysis have revealed a meiotic nature of spores, recombination of genetic material in the process of sporulation and the chromosomal nature of some mutations. A possibility to construct a genetic map of the yeast Pichia pinus MH4 is demonstrated on the basis of tetrad analysis. Three linkage groups are revealed. The life cycle in a homothalic haploid yeast, Pichia pinus, was demonstrated. They are capable to form zygotes and meiotic spores under conditions preventing vegetative growth.     

Transketolase mutation in riboflavin-synthesizing strains of Bacillus subtilis

Unlike its predecessors B. subtilis rosR and 41, riboflavin producing B. subtilis 24 strain does not utilize pentose and gluconate and poorly assimilates glucose. Simultaneous addition of glutamic and shikimic acid restored its capacity to grow and produce riboflavin in medium with pentose and gluconate. This strain lacks the activity of transketolase, the key enzyme of the pentose phosphate cycle, and possesses normal ribulose-5-phosphate-epimerase and glucose phosphate isomerase activities. Like enterobacteria, B. subtilis has two different transport systems for glucose and mannose. The data are discussed from the viewpoint of increasing riboflavin production by transketolase mutants. Probable consequences of cell wall and cytoplasmatic membrane damage in B. subtilis with this mutation are discussed.     

PO-independent termination of transcription of catabolite operons in Escherichia coli and Bacillus subtilis]

The review discusses the Po-independent antitermination in E. coli and B. subtilis. The functional role of antiterminators BglG (in E. coli), SacY, SacT, BglG, LicT, and GlcT (in B. subtilis) is described. Special attention is paid to the role of antiterminators in connection with the phosphoenolpyruvate-dependent phosphotransferase system involved in the carbohydrate transport in bacteria.     

Isolation and mapping of a mutation leading to damage in the cytoplasmic-specific component of the fructose transport system in Escherichia coli K-12 bacteria

A mutant delta ptsH of Escherichia coli was used to obtain a mutation which damages a function of cytoplasmic specific component of the fructose phosphotransferase system--FPr protein. This mutation was mapped using a number of genetical methods. In conjugational crosses the mutation was localized at 47 min of the E. coli chromosomal map. The gene responsible for this defect was designated fpr. In P1 mediated transduction and in conjugational three-factorial crosses the order of the markers in this region was established as: gyrA-ompC-fpr-ptsF-...-his. The frequency of cotransduction of the fpr gene was 4.5 and 35% with gyrA and ompC markers, respectively. In fructose containing minimal medium the doubling time of the ptsH+fpr mutant was lower than that of the delta ptsHfpr+ mutant. Also, the doubling time of the fpr mutant depends on concentration of fructose in growth medium but is independent of the amount of this sugar in the case of the delta ptsH mutant.