Detoxification of Salmonella typhimurium lipopolysaccharide by ionizing radiation

The efficiency of ionizing radiation in detoxifying the lethal determinant(s) of the lipopolysaccharide (LPS) of Salmonella typhimurium, S. enteritidis, and Escherichia coli in aqueous solution and associated with heat-killed S. typhimurium cells in suspension decreased with doses above 1 Mrad. The 50% end point of inactivation was more than 7.0 Mrad for heat-killed salmonellae and 4.8, 4.5, and 1.0 Mrad for the LPS of S. typhimurium, S. enteritidis, and E. coli, respectively. After exposure to 20 Mrad, S. typhimurium LPS retained a small portion of its lethal properties although the ld(50) was much greater than 9.5 mg per 20-g mouse. However, at -184 C, no inactivation of the lethal determinant(s) occurred after exposure to as much as 20 Mrad. This demonstrated the significance of the indirect effect and the mobility and formation of free radicals. At 22 C, the optical density at 400 mmu increased and the pH decreased with increasing radiation dose, but no qualitative changes were observed in the infrared spectrum. No change was observed in the pyrogenicity of S. typhimurium LPS; a slight decrease in antigenicity was revealed when 6 days, but not when 1 day, elapsed between vaccination and challenge in the mouse protection test. The results were interpreted as evidence of the existence of two or more lethal and antigenic determinants. The differential effect of radiation on these properties and on the pyrogenic component(s) probably are indicative of separate functional sites for lethal, antigenic, and pyrogenic activities.     

Study of mutagenic activity of fullerene and some of its derivatives using His+ reversions of Salmonella typhimurium as an example

The influence of three new derivatives of fullerence C60 ([61]dimethoxyphosphoryl[61]carbethoxy-methano[60]fullerene, [61]-(dimethoxyphosphoryl-[61]-carbmethoxy-methanofullerene, and 1-methyl-2-(3,5-di-tertbutyl-4-hydroxy-phenyl)-3,4-fulleropyrrolidine) on the appearance of His+ reversions in the Salmonella typhimurium strain BA13 was studied. It was ascertained that the effect of fullerene derivatives on the occurrence of mutations depends on the type of the molecular group with which fullerene interacts. The biological effect is determined not only by the action of the group associated with fullerene. The dependence between the mutagenic effect and properties of the solvents was detected. Exposure to visible light of the culture treated with fullerene derivatives was found to have an antimutagenic effect in the case of [61]dimethoxyphosphoryl[61]carbethoxy-methanofullerene[60]. For two other derivatives, the differences between experimental and control variants were statistically nonsignificant.     

DNA damage caused by ionizing radiation.

A survey is given of continuous-time Markov chain models for ionizing radiation damage to the genome of mammalian cells. In such models, immediate damage induced by the radiation is regarded as a batch-Poisson arrival process of DNA double-strand breaks (DSBs). Enzymatic modification of the immediate damage is modeled as a Markov process similar to those described by the master equation of stochastic chemical kinetics. An illustrative example is the restitution/complete-exchange model. The model postulates that, after being induced by radiation, DSBs subsequently either undergo enzymatically mediated restitution (repair) or participate pairwise in chromosome exchanges. Some of the exchanges make irremediable lesions such as dicentric chromosome aberrations. One may have rapid irradiation followed by enzymatic DSB processing or have prolonged irradiation with both DSB arrival and enzymatic DSB processing continuing throughout the irradiation period. Methods for analyzing the Markov chains include using an approximate model for expected values, the discrete-time Markov chain embedded at transitions, partial differential equations for generating functions, normal perturbation theory, singular perturbation theory with scaling, numerical computations, and certain matrix methods that combine Perron-Frobenius theory with variational estimates. Applications to experimental results on expected values, variances, and statistical distributions of DNA lesions are briefly outlined. Continuous-time Markov chains are the most systematic of those radiation damage models that treat DSB-DSB interactions within the cell nucleus as homogeneous (e.g., ignore diffusion limitations). They contain virtually all other relevant homogeneous models and semiempirical summaries as special cases, limiting cases, or approximations. However, the Markov models do not seem to be well suited for studying spatial dependence of DSB interactions, which is known to be important in some situations.     

Influence of ionizing radiation on oxygen profiles in different types of multicellular spheroids

Human glioma (U-118 MG and U-138 MG), human colorectal adenocarcinoma (HT-29), human thyroid carcinoma (HTh 7), and hamster embryonic lung (V79-379A) spheroids were irradiated with either single doses of 16 or 40 Gy or fractionated doses of eight times 5 Gy. Oxygen profiles in the spheroids were measured with microelectrodes at different times following irradiation, and these profiles were then compared with the oxygen profiles measured in parallel cultured nonirradiated spheroids. No significant radiation-induced changes in the oxygen profiles were seen in any of the spheroids within the first few days after irradiation. The glioma spheroids did not show any significant increase in oxygen tension even after longer times; however, they were growth inhibited, and the number of S-phase cells was strongly suppressed. Increases in oxygen tension did occur in the HT-29 and V79-379A spheroids but only appeared more than a week after irradiation, when degeneration had started. Histological changes and decrease in diameter were seen in the spheroids that started to degenerate about 5 days after irradiation. Thus radiation doses in the therapeutic range did not, for the spheroids studied, produce rapid increases in the oxygen tension. When a change occurred, it appeared rather late and was probably a consequence of cell degeneration.     

Osteomyelitis and arthritis caused by Salmonella typhimurium in a crow.

Salmonella typhimurium was isolated from an arthritic elbow joint of a crow (Corvus brachyrhynchos) which also had bilateral osteomyelitis of proximal tibias. The prevalence of Salmonella organisms in wild birds is reviewed briefly.     

Spontaneous, ultraviolet and ionizing radiation mutagenesis in two auxotrophic strains of Salmonella typhimurium carrying an R plasmid.

Ultraviolet-induced, gamma-induced and spontaneous mutation yields were studied in two different auxotrophic strains of Salmonella typhimurium in the presence and absence of the UV-protecting drug resistance transfer factor R-Utrecht. One strain, carrying the hisC527 (amber) mutation, showed significantly increased spontaneous, UV- and gamma-induced mutability in the presence of the R-Utrecht plasmid. The other strain, carrying the trpD1 mutation (thought to be a missense mutation), also showed significantly increased UV mutability in the presence of the R-Utrecht plasmid. The other strain, carrying the trpD1 mutation (thought to be a missense mutation), also showed significantly increased UV mutability in the presence of the R factor, but appeared to show no significant increase in spontaneous mutability and only a very slight increase in gamma-mutability when carrying the R factor. These results demonstrate that the R-Utrecht plasmid, known to enhance UV-induced mutation yields in S. typhimurium, can also significantly enhance both spontaneous and gamma-induced mutation yields in this species. The latter effects are not so discernible with all markers, however, as shown by the results with strains carrying the trpD1 mutation. Enhancement of spontaneous mutability thus appears to be correlated with enhancement of gamma-mutability rather than UV mutability.     

Biofilm formation by different strains of Salmonella typhimurium in artificial systems

The ability of 14 different strains of Salmonella typhimurium to biofilm formation depending on genotype and culture conditions was investigated in artificial systems: in 96-well plastic microtitre plates, plastic and glass tubes, plastic Petri dishes and on microscope glasses. Quantitative biofilm growth was monitored by using an assay based on crystal violet staining, while planctonic growth in the same cultures was monitored by absorbance in iEMS Reader MF, and qualitatively--by digital photo and visually. Optimal rate between growth and biofilm indications for all strains was determined at initial cell concentration 10(6-7) KOE/ml and culture incubation at t degrees 28 degrees C. The nutrient content of the medium significantly influenced the quantity of produced biofilm. The nutrient broth LB without NaCl was more effective in promoting biofilm formation, than LB itself. The least quantity of biofilm was formed in water. The genotype of the strains also critically influenced the quantity of produced biofilm. Nonmotile mutants cells had reduced ability to form biofilm. RpoS mutant cells produced significantly less biofilm as compared with cells of isogenic parent strains. The chemical content of plastic and glass also influenced biofilm formation.     

Trends in the development of the epidemic process in salmonellosis caused by Salmonella typhimurium and Salmonella enteritidis

The dynamics and structure of the epidemic process of Salmonella infections among the population of Perm in 1983-1988 was studied and the results of evaluation of antibiotic resistance of the dominating Salmonella species analyzed. The study revealed that a decrease in salmonellosis morbidity caused by S. typhimurium was associated with a limited circulation of anthroponotic (antibiotic-resistant) variants of Salmonellae and a relative increase in the proportion of zoonotic (antibiotic-sensitive) strains. At the period of elevated morbidity this Salmonella infection affected mainly young children in cold months, whereas in recent years seasonal morbidity rises shifted to spring-summer and summer-autumn months, affecting older age groups of the population. The study also revealed that a rise in salmonellosis morbidity caused by S. enteritidis was due to increased circulation of zoonotic variants of Salmonellae. Changes in the epidemiological situation necessitate correction of the system of epidemiological surveillance on Salmonella infections with the emphasis on sanitation measures in stock-breeding farms with unfavorable epidemiological situation.     

The destruction of Salmonella typhimurium in chicken exudate by different freeze-thaw treatments

Antibacterial treatments for frozen poultry, including holding at -5°C and slow thawing at 4°C to which exponential phase cells of Salmonella typhimurium were susceptible, were found to be relatively ineffective against stationary phase cells. Exposure of the latter, however, to a pre-freezing triple stress treatment of cold-shock exposure at 5°C to a solution containing 5 mg/l of free available chlorine in 1% succinic acid (pH 2.5) for 20 min substantially lowered the resistance of the cells to subsequent freezing, storage and thawing in poultry flesh exudate. Cell survival was further decreased by storage of exudate at - 18°C for 28 d and this reduced the proportion of stationary phase cells to less than 1% of initial numbers, with a concomitant increase in sensitivity to deoxycholate. Such a combined pre-treatment may have practical potential for salmonella decontamination in the production of frozen poultry.     

The destruction of Salmonella typhimurium in chicken exudate by different freeze-thaw treatments

Antibacterial treatments for frozen poultry, including holding at -5 degrees C and slow thawing at 4 degrees C to which exponential phase cells of Salmonella typhimurium were susceptible, were found to be relatively ineffective against stationary phase cells. Exposure of the latter, however, to a pre-freezing triple stress treatment of cold-shock exposure at 5 degrees C to a solution containing 5 mg/l of free available chlorine in 1% succinic acid (pH 2.5) for 20 min substantially lowered the resistance of the cells to subsequent freezing, storage and thawing in poultry flesh exudate. Cell survival was further decreased by storage of exudate at -18 degrees C for 28 d and this reduced the proportion of stationary phase cells to less than 1% of initial numbers, with a concomitant increase in sensitivity to deoxycholate. Such a combined pre-treatment may have practical potential for salmonella decontamination in the production of frozen poultry.     

Modeling hematopoietic system response caused by chronic exposure to ionizing radiation

A new model of the hematopoietic system response in humans chronically exposed to ionizing radiation describes the dynamics of the hematopoietic stem cell compartment as well as the dynamics of each of the four blood cell types (lymphocytes, neutrophiles, erythrocytes, and platelets). The required model parameters were estimated based on available results of human and experimental animal studies. They include the steady-state number of hematopoietic stem cells and peripheral blood cell lines in an unexposed organism, amplification parameters for each blood line, parameters describing proliferation and apoptosis, parameters of feedback functions regulating the steady-state numbers, and characteristics of radiosensitivity related to cell death and non-lethal cell damage. The model predictions were tested using data on hematological measurements (e.g., blood counts) performed in 1950–1956 in the Techa River residents chronically exposed to ionizing radiation since 1949. The suggested model of hematopoiesis is capable of describing experimental findings in the Techa River Cohort, including: (1) slopes of the dose–effect curves reflecting the inhibition of hematopoiesis due to chronic ionizing radiation, (2) delay in effect of chronic exposure and accumulated character of the effect, and (3) dose-rate patterns for different cytopenic states (e.g., leukopenia, thrombocytopenia).     

Antioxidative effects of melatonin in protection against cellular damage caused by ionizing radiation

Ionizing radiation is classified as a potent carcinogen, and its injury to living cells is, to a large extent, due to oxidative stress. The molecule most often reported to be damaged by ionizing radiation is DNA. Hydroxyl radicals (*OH), considered the most damaging of all free radicals generated in organisms, are often responsible for DNA damage caused by ionizing radiation. Melatonin, N-acetyl-5-methoxytryptamine, is a well-known antioxidant that protects DNA, lipids, and proteins from free-radical damage. The indoleamine manifests its antioxidative properties by stimulating the activities of antioxidant enzymes and scavenging free radicals directly or indirectly. Among known antioxidants, melatonin is a highly effective scavenger of *OH. Melatonin is distributed ubiquitously in organisms and, as far as is known, in all cellular compartments, and it quickly passes through all biological membranes. The protective effects of melatonin against oxidative stress caused by ionizing radiation have been documented in in vitro and in vivo studies in different species and in in vitro experiments that used human tissues, as well as when melatonin was given to humans and then tissues collected and subjected to ionizing radiation. The radioprotective effects of melatonin against cellular damage caused by oxidative stress and its low toxicity make this molecule a potential supplement in the treatment or co-treatment in situations where the effects of ionizing radiation are to be minimized.     

Ultraviolet-induced reversions of Salmonella his frameshift mutations

Ultraviolet (UV) light elicits reversions in 12 his frameshifts that also revert in response to ICR191. Reversion also is stimulated by UV in two of four spontaneously revertible, but ICR-negative, frameshifts.     

Rapid proteomic remodeling of cardiac tissue caused by total body ionizing radiation

Accidental nuclear scenarios lead to environmental contamination of unknown level. Immediate radiation‐induced biological responses that trigger processes leading to adverse health effects decades later are not well understood. A comprehensive proteomic analysis provides a promising means to identify and quantify the initial damage after radiation exposure. Early changes in the cardiac tissue of C57BL/6 mice exposed to total body irradiation were studied, using a dose relevant to both intentional and accidental exposure (3 Gy gamma ray). Heart tissue protein lysates were analyzed 5 and 24 h after the exposure using isotope‐coded protein labeling (ICPL) and 2‐dimensional difference‐in‐gel‐electrophoresis (2‐D DIGE) proteomics approaches. The differentially expressed proteins were identified by LC‐ESI‐MS‐MS. Both techniques showed similar functional groups of proteins to be involved in the initial injury. Pathway analyses indicated that total body irradiation immediately induced biological responses such as inflammation, antioxidative defense, and reorganization of structural proteins. Mitochondrial proteins represented the protein class most sensitive to ionizing radiation. The proteins involved in the initial damage processes map to several functional categories involving cardiotoxicity. This prompts us to propose that these early changes are indicative of the processes that lead to an increased risk of cardiovascular disease after radiation exposure.