Analysis of mutational specificity induced by heterocyclic amines in the lacZ gene of Escherichia coli

Mutational spectra induced by different heterocyclic amines were characterized and compared with those obtained from diethylnitrosamine and N-methyl-N-nitrosourea. Mutation classes were identified by means of a series of mutant lacZ genes in F' episomes in Escherichia coli engineered to detect specifically each of two transitions, four transversions and five kinds of frameshift events. More than 99.5% of the mutations induced by heterocyclic amines were frameshift mutations. -2(C.G-G.C) frameshifts were favored over other types, such as +1(G.C), -1(G.C), +1(A.T) and -1(A.T), except when 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) was administered. -1(G.C) and +1(G.C) frameshifts predominate following Trp-P-1 treatment. A small number of G.C-->T.A transversions were induced by the treatment with 2-amino-3,4-dimethylimidazo[4,5-f]quinoline as well as with several other heterocyclic amines examined. Since G.C-->T.A transversions, but not frameshift mutations, are reported to play a role in heterocyclic amine-induced activation of the c-Ha-ras protooncogene or inactivation of the p53 tumor suppressor gene, the low level of base substitutions, particularly G.C-->T.A transversions, may represent a partial explanation for the relatively modest carcinogenic activity of heterocyclic amines, despite their extraordinarily strong mutagenicity in the Salmonella mutation assay.     

Development of 170 GHz/500 kW gyrotron

A development of 170GHz/500kW level gyrotron was carried out as R&D work of ITER. The oscillation mode is TE_31,8. In a short pulse experiment, the maximum power of 750kW was achieved at 85kV/40A. The efficiency was 22%. In the depressed collector operation, 500kW/36%/50ms was obtained. The maximum efficiency of 40% was obtained at P_RF=470kW whereas the power decrease by the electron trapping was observed. Pulse extension was done up to 10s at P_RF=170kW with the depressed collector operation. The power was limited by the temperature increase of the output window.     

Antibacterial actin of vinegar against food-borne pathogenic bacteria including Escherichia coli O157:H7 (Part 2). Effect of sodium chloride and temperature on bactericidal activity

Bactericidal effects of various kinds of AWASEZU (processed vinegar, 2.5% acidity) on food-borne pathogenic bacteria including Escherichia coli O157:H7 and other bacteria were examined. the order of bactericidal activities was NIHAIZU (3.5% NaCl was added) > SANBA-IZU (3.5% NaCl and 10% sucrose were added) > plain vinegar (spirit vinegar) > AMAZU (10% sucrose was added). This indicates that their activities were enhanced by the addition of sodium chloride and suppressed by the addition of sugar. On the other hand, when soy sauce was used instead of sodium chloride, the order of bactericidal activities was plain vinegar > AMAZU > NIHAIZU > SANBAIZU. This is mainly because their activities were suppressed by the increase in the pH value. The effect of sodium chloride (0.01-15%) and temperature (10-50 degrees C) on bactericidal activities against E. coli O157:H7 in spirit vinegar (0.5-2.5% acidity) was further examined. When vinegar was used in combination with sodium chloride, predominant synergism on the bactericidal activity was observed. Their activities were markedly enhanced by the addition of sodium chloride in proportion to the concentration. In addition to this, at higher temperatures spirit vinegar killed bacteria much more rapidly. It should be noted that the bactericidal activity of spirit vinegar was extremely enhanced by the combined use of the addition of sodium chloride and the rise of temperature. For example, in 2.5% acidity vinegar, the time required for 3 log decrease in viable cell numbers at 20 degrees C was shortened to 1/140-fold by the addition of 5% sodium chloride, shortened to 1/51-fold by the rise of the reaction temperature at 40 degrees C, and shortened to 1/830-fold; 0.89 minutes by both the addition of 5% sodium chloride and the rise of temperature at 40 degrees C. In order to propose the methods to prevent food poisoning by bacterial infection, bactericidal activities of vinegar solution containing sodium chloride on cooking tools and raw vegetables were examined. Vinegar solution (1-2% acidity, 3-7% NaCl) produced more than 3 log decrease in viable cell numbers of E. coli O157:H7 on the surface of cutting board, and cabbage and cucumber at 20-50 degrees C. These results suggested that the treatment with vinegar solution containing sodium chloride may be one of the useful methods to prevent food poisoning.     

Antigenic and genetic diversities of Babesia ovata in persistently infected cattle

Exploring the antigenic and genetic diversities of Babesia ovata, we obtained several field isolates from grazing cattle in the Okushiri island, Japan. Parasite isolation was greatly facilitated by using bovine red blood cell-substituted SCID mice (Bo-RBC-SCID mice), into which the blood samples of the cattle were inoculated. Isolates from different individuals within a herd of cattle were compared in immunoblot analysis with an anti-B. ovata serum and also in Southern blot analysis with a probe for the small subunit ribosomal RNA gene. In both analyses, the isolates exhibited banding patterns that were significantly different from each other. We were also able to obtain a series of parasite isolates from a single cow in different seasons of a nine months period, including winter when active vector ticks were not in the field environment. Different seasonal isolates showed different banding patterns in both immunoblot and Southern blot analyses. By contrast, these analyses detected little difference among the parasites that had been passed various times in Bo-RBC-SCID mice, where no specific immune responses should be generated. These results indicate that individual animals within a herd of cattle were infected with antigenically and genetically diversified populations of B. ovata, and that the parasites could persistently infect a single animal with dynamic change in their predominant subpopulations.     

Development of a Trans-Mucosal Controlled-Release Device for Systemic Delivery of Antianginal Drugs Pharmacokinetics and Pharmacodynamics

Oral mucosa is well-known to be one of the best routes for drug absorption. But very few R & D works have been initiated to investigate the feasibility of using this site to control drug delivery. A transmucosal controlled-release device, which is capable of achieving excellent absorption and controlled release of drugs, has been developed. The device is a tabletshaped mucoadhesive system which is composed of two layers. The upper layer is a fast-release layer and the lower layer is a sustained-release layer, and designed to be applied between buccal and gingival mucosae. Both layers are formulated from synthetic polymers to control the release of drugs.

Isosorbide dinitrate(ISDN), a well-documented antianginal drug, is known to be susceptible to extensive presystemic elimination when taken orally. It was used as the candidate drug and the systemic bioavailability was studied in human and observed to be improved by as much as 5 fold when compared to a marketed oral sustained-release tablet; On the other hand, much smaller amount of metabolites was formed. The plasma profile of ISDN has also been observed to be substantially prolonged (12 hrs as compared to less than 1 hr for sublingual tablet and spray product on the market). These observations have demonstrated that this device is capable of not only bypassing hepatic “first-pass” metabolism but also having a sustainedrelease property of prolonging the release of ISDN.

Clinical studies performed in the anginal patients for up to one year have demonstrated the therapeutic benefits of this device in achieving a substantial reduction in the frequency of anginal attacks.

This type of device was also applied to the systemic delivery of another antianginal drug, Nifedipine, by employing a formulation with longer sustained drug release property. Again, the clinical results demonstrated that a prolonged duration of therapeutic plasma concentration has also been accomplished.     

Mechanical stress control in a VLSI-fabrication process: a method for obtaining the relation between stress levels and stress-induced failures

An ideal fabrication process is designed to minimize mechanical stress in semiconductor devices and to improve device reliability. Mechanical stress levels were predicted by in-house simulations supported by a thin-film database. These stress levels were correlated with stress-induced defects by TEM analysis supported by fail bit addressing on matured megabit SRAMs. Amorphous-doped silicon film with various annealing temperatures were used for the gate electrode to change the mechanical stress in devices and to get the direct relationship between predicted stress levels and stress related defects. The authors describe brief guidelines for suppressing dislocations in the small geometry shallow-trench isolation process utilizing this system. Polysilicon thickness in the W-polycide gate electrode is designed to minimize mechanical stress in the gate oxide and to suppress the gate oxide failure in probe and class tests. Moreover, critical stress generates dislocations during post source/drain ion implantation anneal obtained by a ball indentation method. This indicated that lower temperature anneal is effective in suppressing the dislocations. A two-step anneal was introduced to suppress dislocations and to enable higher ion activation.     

REM sleep deprivation potentiates the effects of imipramine and desipramine but not that of clomipramine in the forced swimming test

The human amnestic syndrome associated with lesions of the hippocampus and amygdala is characterized by a selective impairment of recent (explicit, episodic) memory. Benzodiazepine (BZ) treated normal subjects demonstrate similar, marked impairments in episodic memory, but in addition, BZ also induces sedation and inattention. Thus, the amnestic effects of BZ may be secondary to drug-induced sedation. However, when subjects were pretreated with the specific BZ receptor antagonist, flumazenil, the sedative and attentional effects of diazepam were blocked, but a marked impairment in episodic memory still occurred. This demonstrates that, using neuropharmacological methods, it is possible to produce a dissociation of memory impairment from inattention and sedation. Such distinct patterns of cognitive dysfunction may serve as models for clinical cognitive syndromes.