Richtmyer-Meshkov instability: theory of linear and nonlinear evolution.

A theoretical framework to study linear and nonlinear Richtmyer-Meshkov instability (RMI) is presented. This instability typically develops when an incident shock crosses a corrugated material interface separating two fluids with different thermodynamic properties. Because the contact surface is rippled, the transmitted and reflected wavefronts are also corrugated, and some circulation is generated at the material boundary. The velocity circulation is progressively modified by the sound wave field radiated by the wavefronts, and ripple growth at the contact surface reaches a constant asymptotic normal velocity when the shocks/rarefactions are distant enough. The instability growth is driven by two effects: an initial deposition of velocity circulation at the material interface by the corrugated shock fronts and its subsequent variation in time due to the sonic field of pressure perturbations radiated by the deformed shocks. First, an exact analytical model to determine the asymptotic linear growth rate is presented and its dependence on the governing parameters is briefly discussed. Instabilities referred to as RM-like, driven by localized non-uniform vorticity, also exist; they are either initially deposited or supplied by external sources. Ablative RMI and its stabilization mechanisms are discussed as an example. When the ripple amplitude increases and becomes comparable to the perturbation wavelength, the instability enters the nonlinear phase and the perturbation velocity starts to decrease. An analytical model to describe this second stage of instability evolution is presented within the limit of incompressible and irrotational fluids, based on the dynamics of the contact surface circulation. RMI in solids and liquids is also presented via molecular dynamics simulations for planar and cylindrical geometries, where we show the generation of vorticity even in viscid materials.

Asymmetric crossing over in the spontaneous formation of large deletions in the tonB-trp region of the Escherichia coli K-12 chromosome.

The chromosomal tonB gene of Escherichia coli was used as a target for the detection of spontaneous deletion mutations. The deletions were isolated in both recA+ and recA- cells, and mutants carrying large deletions were identified because they also lacked part or all of the trp operon. The frequencies of tonB-trp deletion were 1.79 x 10(-9) and 1.09 x 10(-9) for recA+ and recA- cells, respectively. We analyzed 12 deletions from recA+ and 10 from recA- cells by cloning and direct sequencing. The deletions ranged in size from 5612 bp to 15142 bp for recA+ and from 5428 bp to 13289 for recA- cells. Three deletions from recA+ cells and five deletions from recA- cells were found to have occurred between short sequence repeats at the termini of the deletion, leaving one copy of the repeat in the mutant sequence. Seven deletions from recA+ cells and three deletions from recA- cells did not have repeats at their termini; in these cases, the DNA sequences that are adjacent to the deletion termini in the wild-type are characterized by short (2-4 bp) repeats. From these results, a model is presented for the generation of deletion mutations which involves formation of an asymmetric crossover mediated by repeated sequences of 2- to 4-bp.

DNA sequence changes in mutations in the tonB gene on the chromosome of Escherichia coli K12: insertion elements dominate the spontaneous spectra.

To obtain insight into the nature and mechanisms of spontaneous mutations, Escherichia coli K12 strain TM31 was constructed to determine, by DNA sequencing, the mutational spectrum of the tonB gene on the chromosome. We inserted the chloramphenicol resistant gene 1.6 kb upstream of the tonB gene, thus making it possible to retrieve the mutated tonB gene from the chromosome by shotgun cloning using a drug-resistant marker. The spontaneous mutation frequency in the tonB gene, which was judged by its colicin B-resistant phenotype, is 3-10 x 10(-7). Spontaneous mutations were dominated by large insertions that are identified by DNA sequencing to be IS elements; IS1 dominated, but IS2, IS5, and IS10 were also obtained. In uvrA- strain, transposition of both IS10-R and IS10-L are equally increased, suggesting the interaction of the UvrA protein and IS10 transposition. The base substitutions are the second largest group of mutations, among which G:C-->A:T transition is predominant. Deletions also contribute significantly in wild type with regard to DNA repair and uvrA- strains, but not recA- strain, suggesting that the RecA protein is involved to some extent in deletion formation. Endpoints of these deletions do not always correlate with the presence of repeated sequences, indicating the absence of homologous recombination for deletion formation.

MECHANISMS OF SPONTANEOUS MUTAGENESIS IN ESCHERICHIA COLI.

WE HAVE OBTAINED A SPECTRUM OF MUTATIONS OCCURRING BOTH IN THE SUPF GENE ON THE PLASMID AND IN THE TONB GENE ON THE CHROMOSOME. TWENTY SUPF MUTATIONS WERE ANALYZED. BASE CHANGES COMPRISED 55% OF EVENTS, AND TRANSVERSIONS WERE PREDOMINANT. IS ELEMENTS CONTRIBUTED 30% OF THE EVENTS. ONE OF THE TWO DELETIONS WAS ASSOCIATED WITH THE PRESENCE OF 10 BP DIRECTLY REPEATED SEQUENCES, AND A SECOND ONE DID NOT HAVE REPEATS AT THE JUNCTION. A FRAMESHIFT RESULTED FROM THE LOSS OF C IN A RUN OF 5 C RESIDUES. SIX OUT OF THE 7 TONB MUTATIONS OBTAINED FROM THE WILD-TYPE E. COLI WERE IS ELEMENT INSERTIONS WHICH WERE CLUSTERED WITHIN A 100 BP 5' REGION OF THE GENE INCLUDING THE PROMOTER REGION. THE REMAINING MUTATION WAS A DELETION AND DID NOT HAVE REPEATS AT THE JUNCTION. IN THE RECA56 STRAIN, ALL THE TONB MUTATIONS (6 CASES) WERE CAUSED BY THE IS INSERTIONS THAT OCCURRED DISPERSED THROUGHOUT THE GENE. THERE MAY EXIST AN IS INSERTION "HOT SPOTS" IN TONB. THESE RESULTS PROVIDE DIRECT INFORMATION CONCERNING THE NATURE OF SPONTANEOUS MUTATION ARISING IN E. COLI.

Enhancing effect of oxymetholone, an anabolic steroid, on development of liver cell foci in rats initiated with N-diethylnitrosamine.

The promoting potential of oxymetholone (OXM) administration on development of liver cell foci was investigated in male F344 rats previously treated with N-diethylnitrosamine (DEN). One week after a single injection of DEN (100 mg/kg, i.p.), rats were given OXM at a dietary level of 0.2% for the first 4 weeks and then at a concentration of 0.1% for an additional 35 weeks. All rats were killed at week 40 for histopathological and immunohistopathological examination of liver tissue. The numbers and areas of both clear cell and glutathione S-transferase placental form (GST-P) positive foci were significantly increased in the group treated with DEN and OXM as compared with the respective values for the DEN alone group. The results thus suggested that OXM possesses promoting potential for rat liver carcinogenesis.

Tumor promoting effect of goitrogens on the rat thyroid.

To evaluate the mechanism of the promoting effect of goitrogens on thyroid tumorigenesis, well-known goitrogens having different pharmacologic action, i.e., thiourea, phenobarbital sodium (PB), potassium thiocyanate (KSCN), and 3,4,5,6-tetrachloro-2',4',5',7'-tetraiodo-fluorescein sodium salt (Rose Bengal B, FD&C Red No. 105) (FR105) were administered to the DHPN-initiated and non-initiated F344 male rats in the drinking water for 25 weeks. Remington's iodine deficient diet (I-def) was fed as a positive control. These goitrogens showed significant tumor promoting effect or promoting tendency on the rat thyroids. According to the changes in thyroid morphology and thyroid-related hormone titers observed in the present study, we proposed to classify goitrogens at least into 2 groups, i.e., iodine deficiency-type promoters and the iodine excess-type promoters. The former contains goitrogens inducing TSH-stimulated diffuse goiter composed of uniform follicles with activated tall follicular epithelial cells, such as thiourea, KSCN and PB, and the latter contains goitrogens inducing colloid goiter composed of a mixture of colloid-rich follicles with flat follicular cells and normal-looking follicles with cuboidal follicular cells, such as FR105. This classification may be useful for the risk assessment of goitrogens.

Lack of carcinogenicity of tartrazine (FD & C Yellow No. 5) in the F344 rat.

The carcinogenicity of tartrazine (C. I. Food Yellow No. 4, FD & C Yellow No. 5), a food, drug and cosmetics colouring, was examined in F344 rats. Tartrazine was dissolved in distilled water at levels of 0, 1 or 2%, and groups of about 50 male and 50 female rats were given one of these solutions ad lib. as their drinking-water for up to 2 yr. No toxic lesions specifically caused by tartrazine were detected in any treated group of either sex. Many tumours developed in all groups including the control group, and the organ distribution of these tumours and their histological characteristics were similar to those of the spontaneous tumours that are known to occur in this strain of rats. Except for mesothelioma in males and endometrial stromal polyp in females, there were no significant increases in the incidences of any tumours over those in the corresponding control group. In males, mesotheliomas were found only in the group given 1% tartrazine and the incidence of this lesion was statistically significant (Fisher's exact test) in comparison with the other two groups (P less than 0.02). The incidence of endometrial stromal polyp was also significantly higher among females given the 1% dose than in the controls (P less than 0.05). However, no positive trend was noted in the occurrence of these two tumours using an age-adjusted statistical analysis. Mesothelioma and endometrial stromal polyp are frequently observed spontaneous tumours in this strain of rats, and their incidences in our historical controls are 4.1 and 21.9%, respectively. However in the present study mesothelioma occurred in none of the male control rats and the incidence of endometrial stromal polyp was only 10.6% in the female control group. Moreover, there was no significant difference between the control and treated groups in hyperplastic or pre-neoplastic changes in the mesothelium or endometrium. From these findings, we concluded that the significant increases in the incidences of mesothelioma and endometrial stromal polyp that occurred in the groups given 1% tartrazine were not attributable to tartrazine administration. Thus, it is concluded that tartrazine was not carcinogenic in F344 rats when administered continuously at doses of up to 2% in the drinking-water for up to 2 yr.

Long-term studies on carcinogenicity and promoting effect of phenylbutazone in DONRYU rats.

The carcinogenicity and promoting effect of phenylbutazone were investigated in inbred DONRYU rats. In the carcinogenicity study, both sexes were administered the chemical at dietary levels of 0 (control), 0.125, or 0.25% for 2 years. Toxic lesions were associated with phenylbutazone treatment in the kidney and digestive tract, appearing to have an adverse effect on life expectancy. Various tumors were detected in all groups including the controls. With the exception of pheochromocytoma in the female high-dose group, no statistically significant increase in yield of any tumors, including leukemia, was apparent in the treated groups of either sex when the data were analyzed by Fisher's exact probability and/or chi-square tests. Application of an age-adjusted statistical analysis revealed a slight positive effect regarding the occurrence of pheochromocytomas, neoplastic liver nodules, and leukemias in females. However, these tumors are commonly observed to develop spontaneously in this rat strain, and no such effect was apparent in the male groups. In addition, no differences in incidences of relevant preneoplastic lesions were evident between control and treated groups. Thus phenylbutazone showed no carcinogenic activity in DONRYU rats when given continuously in the diet for 2 years. For the investigation of promoting effect, phenylbutazone was given as a dietary supplement for 2 years subsequent to initiation with N-ethyl-N-nitrosourea or N-propyl-N-nitrosourea. No enhancement of nitrosourea-induced leukemogenesis was apparent, although a slight promoting effect was demonstrated for renal and thyroid tumorigenesis.

Glandular changes associated with the spontaneous interstitial cell tumor of the rat testes.

Testes of untreated F344 and Wistar rats in the control groups of carcinogenicity studies were histologically examined, and the histopathological characteristics and histogenesis of glandular changes in these testes were studied. In 266 testes of 2-year-old F344 rats, 263 had interstitial cell tumors (ICTs) (98.9%) and 39 had glandular changes (14.7%). These glandular changes were also found in 1 out of 38 1-year-old F344 rat testes (2.6%), and 3 in 154 2-year-old Wistar rat testes (1.9%). The changes were observed exclusively in the interstitial cell tumors (ICTs). These glandular changes showed variation in size, shape and number. They were composed of tubules or cysts lined by a layer of cuboidal or columnar epithelial cells, which had terminal bars, occasionally PAS- and alcian blue-positive brush borders, and rarely, alcian blue-positive cytoplasmic vacuoles. Serial sections revealed that the changes were not connected with the rete testes, but with the degenerative seminiferous tubules involved in the ICTs lined by a layer of flat endothelial-like cells. The findings suggest that the lesions constitute metaplastic changes of the Sertoli cells.

Spontaneous neoplastic and non-neoplastic lesions in aging Donryu rats.

Spontaneous neoplastic and non-neoplastic lesions in 95 male and 96 female Donryu rats which were observed up to 120 weeks of age, were examined. The incidence of spontaneous tumors was 73.7% in males and 88.5% in females. In males, the most frequent tumors were pituitary adenomas, followed by pheochromocytomas and insulinomas. In females, uterine adenocarcinomas, mammary fibroadenomas and pituitary adenomas were the most common. Other tumors with relatively high incidences in both sexes included cortical adenomas of the adrenal gland, histiocytic sarcomas of the hematopoietic organs and granular cell tumors of the brain. Various tumors were also found in many other organs and/or tissues, although their incidences were low. The organ distribution and incidences of spontaneous tumors observed in Donryu rats were different from those in other strains of rats such as the ACI, Wistar, F344 or Sprague-Dawley strains. The main non-neoplastic lesions were observed in the lung, cervical lymph nodes and kidney of both sexes. In addition, lesions were also observed in the urinary bladder, prostate and peripheral nerves (spinal nerve roots and peripheral nerves) and/or femoralis muscle of males. Histologically, the most characteristic lesion was radiculoneuropathy with degeneration of the peripheral nerves.

Lack of carcinogenicity of triethanolamine in F344 rats.

The carcinogenic potential of triethanolamine was examined in F344 rats. Triethanolamine was dissolved in distilled water at levels of 0 (control), 1, and 2%, and groups of 50 males and 50 females were given these doses ad libitum as drinking water for 2 yr. The dose levels in females were reduced by half from wk 69, because of associated nephrotoxicity. A variety of tumors developed in all groups, including the control group, and all tumors observed were histologically similar to spontaneous tumors in this strain of rats. No statistically significant increase of the incidence of any tumor was observed in the treated groups of both sexes by the chi-square test. In this study, however, there was an increase in nephrotoxicity, which appeared to have an adverse effect on the life expectancy of the treated animals, especially of females. Therefore, an age-adjusted statistical analysis on incidences of main tumors or tumor groups of both sexes was also done by methods recommended by Peto et al. (1980). The result showed that a positive trend (p less than 0.05) was noted in the occurrence of hepatic tumors (neoplastic nodule/hepatocellular carcinoma) in males and of uterine endometrial sarcomas and renal-cell adenomas in females. These tumors, however, have been observed spontaneously in this strain of rats, and their incidences in the control group of the present study were lower than those of our historical controls. These results may indicate that a positive trend in the occurrence of these tumors is not attributable to triethanolamine administration. Increased incidence of renal tumors in the female high-dose group may have been connected with renal damage. Histological examination of renal damage observed in the treated groups, especially in the female high-dose group, revealed acceleration of so-called chronic nephropathy. In addition, mineralization of the renal papilla, nodular hyperplasia of the pelvic mucosa, and pyelonephritis with or without papillary necrosis were also observed. Thus, it is concluded that under these experimental conditions triethanolamine is not carcinogenic in F344 rats but is toxic to the kidneys.

Relation between development of leukemia and duration of N-nitroso-N-ethylurea treatment in DONRYU rats.

N-Nitroso-N-ethylurea (NEU; CAS: 759-73-9) is a strong leukemogen that induces erythroblastic leukemia in inbred DONRYU rats. In the present experiments, relationships between development of leukemia, duration of NEU treatment, and sequential changes in the hematopoietic organs during carcinogen administration were examined. In experiment 1, groups of rats were given a 400-ppm NEU solution for 0, 2, 4, 6, 8, or 10 weeks, and the resultant incidence of leukemias was 0, 26, 40, 75, 95, and 100%, respectively. Of the various types of leukemia, the erythroblastic type was observed in 0, 0, 20, 40, 95, and 90% of rats, respectively. The average latent period showed an inverse correlation with the duration of NEU treatment. In experiment 2 the animals were divided into carcinogen-treated and control groups, and rats were sacrificed periodically for histopathologic examination. In the experimental group, the bone marrow became hypoplastic soon after commencement of NEU treatment and at the 6th week became severely aplastic, thereafter recovering slightly. At the 10th week, 2 rats out of 5 examined were leukemic. Relationships between incidence of leukemia, duration of NEU treatment, and sequential changes of the bone marrow during carcinogen administration are discussed.

Effect of Nocardia rubra cell wall skeleton and/or cyclophosphamide on leukemogenesis induced by N-ethyl-N-nitrosourea in Donryu rats.

The effect of Nocardia rubra cell wall skeleton (N-CWS) and/or cyclophosphamide (CP) on chemical carcinogenesis was examined in female Donryu rats exposed to N-ethyl-N-nitrosourea (ENU) in the drinking water for 6 weeks. Five administrations of N-CWS following ENU treatment caused a slight prolongation of the average survival of rats but did not reduce the incidence of leukemia. CP given on two occasions after ENU treatment caused a moderate prolongation of average survival period and a moderate reduction of the incidence of leukemia, but significant differences from ENU-treated control group values were not observed after statistical analysis. Combined treatment with N-CWS and CP after ENU treatment caused prolongation of the average survival period of rats and a statistically significant reduction in the incidence of leukemia. The present experiment indicates that combined treatment with N-CWS and CP effectively reduces induction of leukemia by ENU in rats, although other types of tumors were not affected.

Organ-specific carcinogenicity of N-methyl-N-nitrosourea in F344 and ACI/N rats.

Male and female F344 rats were continuously administered N-methyl-N-nitrosourea (MNU) in their drinking water at concentrations of 200 or 100 ppm, and both sexes of ACI/N rats were given MNU at a concentration of 200 ppm. By the 42nd week of the experiment, high incidences of brain/spinal cord tumors were observed in both strains of rats. Histologically, many of them were astrocytomas or anaplastic astrocytomas. In addition, malignant neurinomas were also detected in the spinal nerve roots and trigeminal nerves, although their incidences were rather low. There was no difference in the type and incidence of these neurogenic tumors between the two strains of rats. Tumors of the tongue and esophagus were mainly observed in the high-dose group of F344 rats and those of the glandular stomach were observed in the low-dose group of F344 rats. In ACI/N rats, tumors of the heart and renal pelvis were detected. The organ-specific carcinogenicity of MNU in these two strains of rats was compared with that of MNU in Donryu rats. It was demonstrated that organ specificity of MNU given orally was influenced not only by the strain of rats but also by the dose level.

Development of thymic lymphomas by oral administration of N-nitroso-N-propylurea and establishment of transplantable lines of thymic lymphoma in F344 rats.

Forty-eight female F344/DuCrj rats were given a 400-ppm solution of N-nitroso-N-propylurea (CAS: 816-57-9) continuously in their drinking water. Thymic lymphomas were induced most frequently (85%) followed by duodenal tumors (48%). Sixteen tumors were examined by the immunofluorescence inhibition test for murine leukemia virus-related antigens; 15 were negative and the other was weakly positive. Twenty-six tumors were intraperitoneally and subcutaneously transplanted syngeneically; 25 (96%) were successfully transplanted intraperitoneally and 24 (92%) subcutaneously. The serial intraperitoneal transplantation was continued, and 22 lines of transplantable lymphoma in an ascites form were established. In almost all tumor lines, tumor cells took in a high percentage of recipient rats and caused the death of the recipients within 12-23 days. The thymus, liver, spleen, greater omentum, and lymph nodes were frequently invaded by transplanted tumor cells. The tumor lines were considered to be of T-cell lineage.

Foreign-body tumorigenesis in rats by various kinds of plastics--induction of malignant fibrous histiocytomas.

Five kinds of plastics (3 polyvinyl chlorides, 1 polyhydroxyethyl metacrylate and 1 dimethyl polysiloxane) were implanted into subcutaneous tissues of Wistar rats of both sexes. Subcutaneous tumors developed in all experimental groups. The incidences of the tumors, however, differed from each other, although these materials were tested on the same experimental condition. This result indicates that chemical characters of the materials may influence the incidence of subcutaneous tumors. Histologically, most of these subcutaneous tumors were mesenchymal tumors with spindle cells arranged in a storiform pattern, with sheets of histiocyte-like cells or pleomorphic giant cells. Electron microscopy showed mixture of fibroblastic cells, histiocytic cells and undifferentiated cells in these tumors. From these histological and electron microscopical findings, many of the tumors were diagnosed as malignant fibrous histiocytomas.

Carcinogenicity of low doses of N-ethyl-N-nitrosourea in F344 rats; a dose-response study.

The threshold level of minimum carcinogenic dose of N-ethyl-N-nitrosourea (ENU) and the effect of dose on organ specificity were examined by continuous oral administration to both sexes of F344 rats of low doses of ENU at four concentrations (0.3, 1, 3 and 10 ppm) in the drinking water. ENU at 10 ppm selectively induced neurogenic tumors and tumors of the digestive tract, including duodenal tumors. Even at lower doses it enhanced the spontaneous development of some other tumors. A high dose of ENU (400 ppm) was previously found to induce duodenal tumors selectively. These results indicate that the organ specificity of ENU is influenced by the dose and that ENU has multi-potent carcinogenic activity in many organs and/or tissues. In this study, some specific tumors, such as those of the nervous system and digestive tract, seemed to require a minimum carcinogenic level of ENU (10 ppm) for their appearance. However, it seems that ENU is carcinogenic at much lower dose levels than 10 ppm because ENU enhanced the spontaneous development of some other tumors in many experimental groups. The so-called virtually safe doses inducing these specific tumors at a risk level of 10(-6) were calculated.

Malignant fibrous histiocytomas induced in rats by polymers.

Five polymeric materials (3 polyvinyl chlorides, 1 polyhydroxyethyl methacrylate, and 1 dimethyl polysiloxane) were implanted into subcutaneous (SC) tissues of rats. Subcutaneous tumors developed in all experimental groups. The incidences of the tumors differed however, although the experimental conditions were the same for all these materials. This result indicates that chemical characteristics of the materials may influence the incidence of SC tumors. From the histological and electron-microscopic findings many of these tumors were diagnosed as malignant fibrous histiocytomas.

Induction of digestive-tract tumors in F344 rats by continuous oral administration of N-butyl-N-nitrosourea.

Male and female F344/DuCrj rats were administered N-butyl-N-nitrosourea at a concentration of 400 ppm in their drinking water. By the 50th week of the experiment, the cumulative incidence of upper-digestive-tract tumors was as high as 35/39 (90%) and 34/39 (87%) in male and female rats, respectively. Among these, esophageal and forestomach tumors occurred most frequently. Except one female rat with fibroma, upper-digestive-tract neoplasms were of the epithelial type -- papilloma, squamous-cell carcinoma or adenocarcinoma. In female rats, vaginal tumors were induced in 16 (41%) animals. Ear-duct tumors and hematopoietic neoplasms were also induced in both sexes of rats, with incidence of less than 21%.