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Characteristic bacteriolytic activities of Staphylococcus hyicus.

Staphylococcus hyicus demonstrated characteristic bacteriolytic activities towards a Micrococcus luteus reference strain. This lytic activity was demonstrated on medium containing M. luteus cells as large zones of transparency around the culture streak. Smaller zones of transparency were observed with Staphylococcus intermedius, Staphylococcus chromogenes, and some strains of Staphylococcus aureus but not with other coagulase-negative staphylococcal species. The distribution and extent of the bacteriolytic activity could be useful as additional criteria for differentiation of S. hyicus.

Animals↗

Inhibition of DNA gyrase by optically active ofloxacin.

Inhibition of DNA gyrase activity by optically active ofloxacins was studied and compared with the inhibition of norfloxacin and ciprofloxacin. The (-)-isomer of ofloxacin inhibited the supercoiling activity of gyrase from Micrococcus luteus more effectively than did the (+)-isomer. The 50% inhibitory concentrations of (-)-, (+/-)-, and (+)-ofloxacin; norfloxacin; and ciprofloxacin for gyrase from Escherichia coli were 0.78, 0.98, 7.24, 0.78, and 1.15 microgram/ml, respectively. These values correlated well with the antibacterial activity of each compound against intact bacterial cells.

Anti-Bacterial Agents↗

[Investigations on the standardization of the lysoplate technique for assay of lysozyme].

In this paper studies for the standardization of the estimation of lysozyme in low concentrations concerning the lysoplate technique are reported. The influence of the conditions of diffusion, the concentration and the age of the bacteria as well as the judgement of different pictures of lysis and contrast resp. have been measured and compared. Sufficient results have been obtained with "Purified agar" (Difco) in a concentration of 0.9% and with living, 48 h old strains from Micrococcus luteus. After this a mixture of the bacteria in a concentration of 2.75 x 10(8)/ml with 5 ml "Purified agar" at 60 degrees C can be recommended. Using conditions of incubation with 50 degrees C and a final time of 4 h lysozyme can be estimated certainly with a sensitivity of 1 microgram/ml.

Agar↗

Evidence for an increased level of DNA damage in high doubling level human diploid cells culture.

Excision repair after ultraviolet (UV) irradiation was estimated in human diploid fibroblast-like cells (HDF) by treating DNA with a crude extract from Micrococcus luteus that contained pyrimidine dimer-specific endonucleases. The assays were done before and after the cells had been arrested in an essentially nonmitotic state. When low and high population doubling level (PDL) cells were assayed under these conditions, no age-related difference in the number of UV-induced endonuclease sensitive sites was observed, but the removal of these sites was more rapid in high PDL arrested cells. There was a difference between the calculated number average molecular weights (Mn) of DNA from unirradiated low and high PDL arrested cells. The lower Mn of the DNA from high PDL cells indicated that other enzymes present in the M. luteus extract were acting upon non-UV-induced DNA distortions and that these were present to a greater extent in the chromatin associated regions of older cells. These results support the hypothesis that DNA damage accumulates as HDF progress through their in vitro life span.

Cell Division↗

Elemental composition of bacterial metachromatic inclusions determined by electron microprobe X-ray analysis.

Electron microscopy and microprobe X-ray analysis were used to study metachromatic inclusions of Spirillum itersonii , Corynebacterium diphtheriae, and Micrococcus luteus. In situ metachromatic inclusions were electron dense and contained phosphorus and divalent cations. Metachromatic inclusions isolated by anion-exchange column chromatography and by isoosmolar Metrizamide density gradient centrifugation were similar in composition to in situ inclusions.

Calcium↗

In vitro inhibition of bacterial DNA gyrase by cinodine, a glycocinnamoylspermidine antibiotic.

Cinodine, a broad-spectrum glycocinnamoylspermidine antibiotic, binds to DNA and irreversibly inhibits bacterial and phase DNA synthesis. Cinodine was found to inhibit the activity of Micrococcus luteus DNA gyrase in vitro, but it did not inhibit the activities of two other DNA-binding enzymes, namely, topoisomerase I and BamHI. Although we cannot yet conclude that DNA gyrase is an intracellular target of the drug, in vitro inhibition of the enzyme by cinodine appears to be specific.

Aminoglycosides↗

Inverse correlation in nutritionally variant streptococci between the production of bacteriolytic activity and sensitivity to a Streptococcus pyogenes bacteriocinlike inhibitory substance.

Nineteen strains of nutritionally variant streptococci (NVS) were tested for bacteriolytic activity and for their production of and sensitivity to streptococcal bacteriocinlike inhibitory substances (BLIS). None appeared to produce BLIS. An inverse relationship was found between the sensitivity to BLIS and the production of bacteriolytic activity against Micrococcus luteus. All but one of the 14 Streptococcus defectivus isolates were sensitive to the BLIS of S. pyogenes P5, and these isolates were nonlytic. The five S. adjacens isolates were not sensitive to any BLIS tested, and all of these isolates displayed bacteriolytic activity. Sensitivity to the BLIS of S. agalactiae P3 separated the nonlytic S. defectivus strains into two categories.

Bacteriocins↗

Reclassification of Micrococcus agilis (Ali-Cohen 1889) to the genus Arthrobacter as Arthrobacter agilis comb. nov. and emendation of the genus Arthrobacter.

Phylogenetic evidence derived from a 16S ribosomal DNA analysis indicated that the type strain of Micrococcus agilis, DSM 20550 (= ATCC 966 = CCM 2390), is less closely related to the type species of the genus Micrococcus, Micrococcus luteus, than to the type species of the genus Arthrobacter, Arthrobacter globiformis, and related Arthrobacter species. The phylogenetic position of M. agilis is supported by the presence of peptidoglycan variation A3 alpha and by the presence of MK-9(H2) as the major isoprenolog, a characteristic also found in strains of A. globiformis, Arthrobacter crystallopoietes, Arthrobacter atrocyaneus, Arthrobacter citreus, Arthrobacter aurescens, Arthrobacter ilicis, Arthrobacter ureafaciens, Arthrobacter oxydans, Arthrobacter histidinolovorans, and Arthrobacter nicotinovorans. The last six species and M. agilis are characterized by the presence of threonine in the interpeptide bridge of the peptidoglycan. Threonine has not been found in the peptidoglycans of other Arthrobacter species or in members of the genus Micrococcus. Despite the fact that a morphological life cycle is not known, these data support the proposal that M. agilis should be transferred to the genus Arthrobacter as Arthrobacter agilis comb. nov.

Arthrobacter↗

Dissipative structures in proteoglycan solutions.

Diffusion in multicomponent solutions containing proteoglycan is shown to result in the formation of coherent, fluid structures (known as dissipative structures) and induction of rapid polymer transport. These phenomena occur over a wide range of conditions (i.e. varying solute distribution, concentration, size, and chemical composition) which are envisaged to occur in the extracellular matrix of connective tissues. A concentration gradient of chondroitin sulfate in a proteoglycan matrix of uniform concentration yields dissipative structures which transport the proteoglycan up to 300-fold faster than its transport in the absence of the gradient component. Similar behavior was observed with other polysaccharide and monosaccharide concentration gradient components. Amplification of structure formation and rapid transport was achieved by 1) increasing the concentration of proteoglycan matrix, 2) increasing the magnitude of the concentration gradient, 3) decreasing the molecular weight of the gradient-forming component, and 4) decreasing the concentration gradient of proteoglycan in the matrix. Dissipative structure morphology exhibits a marked dependence on the initial component distribution. Non-specific, excluded volume interactions between the proteoglycan and the gradient component are believed to induce coupled diffusive transport of the proteoglycan. This leads to microscopic density inversions which nucleate and develop into macroscopic convective flows. These results are similar to those previously observed in ternary solutions of uncharged polymers (i.e. dextran/polyvinylpyrrolidone). We have demonstrated that dissipative structures may transport Micrococcus luteus cells as well as various solutes. Flows were also observed in proteoglycan solutions after localized addition of small amounts of either a proteolytic enzyme or hyaluronic acid. It is likely that the prerequisites for this spontaneous macroscopic self-organization, as manifested by the flow phenomenon, are present in the extracellular matrix of connective tissues.

Animals↗

Nucleoside diphosphokinase activity associated with DNA polymerases.

Nucleoside diphosphokinase activity is present in highly purified preparations of DNA polymerase from Micrococcus luteus and Escherichia coli, and in a partially purified DNA polymerase from avian myeloblastosis virus. The activity is also observed in the protein fragment of molecular weight 76,000 that is produced by subtilisin cleavage of DNA polymerase I from E. coli. The NDP kinase activity in DNA polymerase preparations from M. luteus uses various ribo- and deoxyribonucleoside di- and triphosphates as substrates. The presence of this activity in preparations of DNA polymerase results in the apparent use of deoxyribonucleoside diphosphates as substrates for DNA synthesis, provided that some triphosphate is present to serve as a phosphate donor.

Adenosine Diphosphate↗

Caffeine inhibits DNA polymerase I from Escherichia coli: studies in vitro.

Caffeine inhibits the activity of DNA polymerase I (E. coli) and its proteolytic large fragment in in vitro DNA replication system. DNA polymerase from Micrococcus luteus is also equally inhibited by caffeine. The extent of inhibition was more with the activated adenovirus, T4 and calf thymus DNA than with synthetic DNA template-primers. Results obtained from time-course studies indicated that caffeine inhibition reached maximum by 30 min of incubation. Enzyme kinetic studies showed that inhibition was competitive with respect to DNA template.

Adenoviridae↗

Nonspecific and cooperative binding of lectins to microorganisms.

Binding of lectins to microbial cell walls was investigated by fluorimetric titration and Scatchard plot. Data were correlated with agglutinability. Concanavalin A and lectins of wheat germ, soybean, pea, lentil, and peanut were tested against Escherichia coli. Micrococcus luteus, Lactobacillus plantarum, and Bacillus subtilis. In cases where binding occurred, it was either nonspecific or positively cooperative. Agglutination was observed only in those combinations of lectin and microorganism that showed positive cooperative binding, suggesting a definite relation between binding and agglutination. Lectins binding to the same carbohydrate did not necessarily bind to the same microorganism, confirming the complexity of the lectin-receptor bond.

Agglutination↗

Variations in excision repair of UVB-induced pyrimidine dimers in DNA of human skin in situ.

The excision repair kinetics of UVB (280-320 nm)*-induced pyrimidine dimers in DNA of human skin in situ was determined for seventeen volunteers using a dimer-specific endonuclease from Micrococcus luteus in conjunction with agarose gel electrophoresis. Removal of pyrimidine dimers from human skin could be detected within 6 h after irradiation and the average half-life for removal of pyrimidine dimers was 11.0 h (+/- 4.3 h). However, there was significant inter-individual variability of repair as indicated by a half-life coefficient of variation of 38%.

Adult↗

High-performance liquid chromatographic determination of cefoperazone in human serum and urine.

A gradient high-performance liquid chromatographic (HPLC) procedure has been developed for the determination of microgram amounts of cefoperazone in human serum and urine. The method employs a muBondapak C18 column and gradient elution with two mobile phases. Excellent separation of the drug from potential degradation products as well as from representative penicillins (sodium ampicillin, sodium methicillin, potassium penicillin G) and aminoglycosides (tobramycin, gentamicin, kanamycin) has been demonstrated. Coefficients of variation of 7.3% or less were obtained for 25-100 micrograms/ml cefoperazone in both serum and urine. Average recoveries of the drug from spiked serum and urine samples corresponded to 97.6% and 98.6%, respectively. Amounts as low as 1 microgram cefoperazone per ml of sample can be estimated using sample volumes corresponding to 0.1 ml serum or 1 ml urine. Good correlation between the HPLC assay and a microbiological cylinder-plate assay employing Micrococcus luteus ATCC 9341 has been demonstrated for human serum and urine of patients treated with cefoperazone. While the microbiological method is less time-consuming, it lacks specificity in the presence of other antibiotics. The HPLC method can be used to analyze cefoperazone in the presence of penicillins and aminoglycosides which can potentially be co-administered with cefoperazone.

Anti-Bacterial Agents↗

Template-primer activity of 5-(hydroxymethyl)uracil-containing DNA for prokaryotic and eukaryotic DNA and RNA polymerases.

We have utilized Bacillus subtilis phage SPO-1 DNA as a model of irradiated DNA. In this phage, all thymine (Thy) residues are replaced by 5-(hydroxymethyl)uracil (5HmUra), which is a known irradiation-induced derivative of DNA Thy. SPO-1 phage is naturally devoid of other such irradiation-induced DNA lesions. DNase I activated SPO-1 phage DNA served as well as, or even better than, the control DNAs (Bacillus subtilis DNA and calf thymus DNA) as a template-primer for Escherichia coli, Micrococcus luteus, and human HL-60 cell DNA polymerases. Furthermore, the template activity of SPO-1 phage DNA was also superior when transcription with E. coli RNA polymerase was investigated. The results reported here indicated that the replacement of Thy by 5HmUra is not deleterious to template and primer functions during DNA or RNA synthesis.

Bacillus subtilis↗

Interactions of Bacillus licheniformis ATCC 10716 and normal flora of human skin.

To determine whether antibiotic production might be ecologically advantageous in the survival of Bacillus species on human skin, we applied spores of a bacitracin-producing strain of Bacillus licheniformis (ATCC 10716) to the forearms of 11 volunteers. Three additional strains of B. licheniformis which did not synthesize antibiotics, including a mutant of ATCC 10716, were used in subsequent control trials. Samples of flora were taken from inoculated and control (opposite forearm) sites during the colonization period, generally 3 weeks. Although population densities were unaltered, changes in the carriage, composition, and bacitracin sensitivity of resident flora were related with the presence of ATCC 10716 only, which suggests that microbial interactions are important in bacillus colonization and in maintenance of normal flora. Interactions were examined in vitro by comparing growth curves of representative skin bacteria, including isolates of Staphylococcus epidermidis, Staphylococcus saprophyticus, Micrococcus luteus, and a large-colony diphtheroid, grown individually, in mixed culture with each other, and together in presence of each test strain of B. licheniformis. We observed some diminution of growth of M. luteus and the diphtheroid in the first mixed culture, and the diphtheroid was completely retarded in common culture with ATCC 10716. Lesser antibiotic effects were seen on the cocci, whose rank of sensitivity was similar to that in vivo. The growth of the diphtheroid was enhanced in mixed culture with those strains of bacilli which lack antibiotic activity.

Adult↗

A comparison of the effects of daunomycin and adriamycin on various DNA polymerases.

The effects of the anthracycline antiboties, daunomycin and adriamycin, on the DNA-directed activities of DNA polymerases from murine sarcoma virus, rat liver (high-molecular-weight species), Escherichia coli, and Micrococcus luteus were determined. Under all conditions tested, these compounds had greater inhibitory effect against the viral polymerase than against cellular polymerase. The inhibition of murine sarcoma virus DNA polymerase by daunomycin was competitive with respect to DNA. For viral DNA polymerase it was concluded that the inhibition was predominatly caused by the interaction of duanomycin with the primer-template DNA. Also, an appreciable reversal of the daunomycin-induced inhibition of this polymerase by an increase in Mg-2+ concentration is consistent with the conclusion derived by competition experiments. In contrast, the inhibition of both rat liver and M. luteus DNA polymerases was essentially noncompetitive with DNA. Also, bacterial enzymes wer e less sensitive to inhibition by these drugs than the virion polymerase. The strong and preferential inhibiton of viral DNA polymerase is discussed in relation to a differential sensitivity of normal as compared to tumor cells observed in some cell lines.

Animals↗

Synthesis and study of antibacterial and antifungal activities of novel 2-[[(benzoxazole/benzimidazole-2-yl)sulfanyl] acetylamino]thiazoles.

Several 2-[[(benzoxazole/benzimidazole-2-yl)sulfanyl]acetylamino]thiazoles derivatives were synthesized by reacting 4-substituted-2-(chloroacetylamino)thiazoles with benzoxazole/benzimidazole-2-thioles in acetone and in the presence of K2CO3. The chemical structures of the compounds were elucidated by IR, 1H-NMR, and FAB(+)-MS spectral data. Their antimicrobial activities against Micrococcus luteus (NRLL B-4375), Bacillus cereus (NRRL B-3711), Proteus vulgaris (NRRL B-123), Salmonella typhimurium (NRRL B-4420), Staphylococcus aureus (NRRL B-767), Escherichia coli (NRRL B-3704), Candida albicans and Candida globrata (isolates obtained from Osmangazi Uni. Fac.of Medicine) were investigated and in this investigation, a significant level of activity was illustrated.

Anti-Bacterial Agents↗