Growth and differentiation potentials in confluent state of culture of human skeletal muscle myoblasts

The transitional behaviors of myoblasts toward differentiation were investigated in the cultures at the low and high seeding densities (respectively, X₀ = 1.0 × 10³ and 2.0 × 10⁵ cells/cm²). In the culture at the low seeding density, an increase in confluence degree accompanied a decrease in growth potential (R_p), being R_p = 0.85 and 0.11 at t = 48 and 672 h, respectively. Myoblasts seeded at the high density resulted in the immediate cessation of growth with keeping the low range of R_p = 0.02–0.09 throughout the culture. The reduction of R_p led to the generation of three subpopulations of cells in proliferative, quiescent and differentiated states. Close cell contacts in the confluent state of high seeding culture induced cell quiescence to a higher extent with suppressing differentiation.     

Effects of additives on prebreakdown phenomena in n-hexane

Prebreakdown phenomena in n-hexane are observed in detail for positive and negative polarities by using simultaneously a high speed schlieren technique and an LED current measuring system, when an impulse voltage (1.1/225 μs) is applied to a point-to-plane electrode gap. Furthermore, the effects of several additives on the streamer propagation are investigated. Especially the effects of electron-trapping additives on negative streamer propagation and of low ionization potential additives on the positive streamer propagation, are examined, as is a correlation between the shape and the propagation velocity of the streamers     

Beating in Membrane Potential Mediated by Cortical γ-Oscillations for Reading Out Synchronous Neuronal Activities Over Distant Cortical Areas

I investigate whether cortical -oscillations are relevant to reading out synchronous neuronal firings that are distributed over the brain. A cortical neural network model is proposed and simulated. The model consists of two sensory networks (SI and SII) and one coincidence-sensitive network (CS). -oscillatory inputs and external stimuli are applied to interneurons and projection neurons of the sensory networks, respectively. The SI and SII networks project to the CS network in a divergent/convergent manner. Here I show that beating in membrane potential of CS neurons mediates the detection of synchronous neuronal firings between the SI and SII networks. A slight increase (a few Hz) in difference between frequencies of the -oscillations in the SI and SII networks generates the beating in the CS network. The synchronous neuronal firings are detected at each peak of the beating, regardless of difference in signal transmission time between SI-to-CS and SII-to-CS pathways.     

In vitro construction and screening of a Burkholderia cepacia lipase library using single-molecule PCR and cell-free protein synthesis

A mutant library of Burkholderia cepacia lipase KWI-56 was constructed on microplates by a cell-free process and tested with a chromogenic assay. This high-throughput construction system can be used to screen mutant proteins based on their catalytic activity.     

The alpha-aminoadipate pathway for lysine biosynthesis is widely distributed among Thermus strains

We previously reported that lysine is synthesized through the alpha-aminoadipate pathway in Thermus thermophilus HB27 (T. Kosuge and T. Hoshino, FEMS Microbiol. Lett., 169, 361-367, 1998), which was the first report demonstrating the synthesis of lysine through the alpha-aminoadipate pathway in Bacteria. LYS20 and LYS4, which respectively encode homocitrate synthase and homoaconitate hydratase have already been identified as the lysine biosynthetic genes in T. thermophilus HB27. In the present work, we examined eight other Thermus strains for the existence of genes belonging to the alpha-aminoadipate pathway. BamHI- or BglII-digested total DNAs from the eight strains were analyzed by Southern hybridization using LYS20 or LYS4 as a DNA probe. DNA fragments that hybridized with one or both of the genes were detected in seven of the Thermus strains but not in T. ruber. The sizes of the fragments that hybridized with the LYS20 and LYS4 probes were the same among T. thermophilus HB27, T. thermophilus HB8, "T. caldophilus" GK24, and four "T. flavus" strains. For example, a similar 4.3-kb fragment was detected in each of the above seven strains. In this fragment, four open reading frames were found downstream of the LYS4 gene in T. thermophilus HB27. Gene disruption experiments revealed that three open reading frames are involved in lysine biosynthesis in T. thermophilus HB27. These results strongly suggest that the lysine biosynthetic gene cluster for the alpha-aminoadipate pathway is widely distributed in the genus Thermus.     

A study on anticoagulation for cardiovascular diseases using international normalized ratio (INR)

The usefulness of INR as an index for assessing anticoagulation was studied using 85 outpatients, all suffering cardiovascular diseases and receiving warfain as subjects. The thrombotest, prothrombin time and INR were measured at follow-up, monthly intervals. The correlation between the thrombotest value and the INR value were assessed retrospectively. We found a stronged correlation between them. The 10 approximately 25% range of the thrombotest value, the therapeutic range, corresponded to 2.29 approximately 3.60 of the INR value. The INR value was more accurate than the thrombotest valve in the situation of prolonged coagulation time. It is recommended to use INR as an objective index for the anticoagulation due to increase accuracy and objectivity when comparing the data among the various institutions.     

Antimicrobial activity of macrolides against clinical isolates

Antimicrobial activity of 6 macrolides was determined using a micro-broth dilution method, against 535 clinical isolates of 22 species, which were isolated in 1996 from 325 facilities in Japan. Results were as follows. 1. In general, antimicrobial activities of 14-membered macrolides were higher than those of 16-membered macrolides. The antimicrobial activities of 14-membered macrolides were in the order of clarithromycin (CAM), erythromycin (EM), roxithromycin (RXM). Among 16-membered macrolides, rokitamycin (RKM) was the most potent, josamycin (JM) was next potent followed by midecamycin (MDM). More numbers of highly-resistant strain of > 100 micrograms/ml were recognized in 14-membered macrolides than in 16-membered macrorides. 2. Most of S. pyogenes (group A) strains were distributed in the susceptible range and almost none was found in the resistant range. 3. S. pneumoniae strains were distributed widely from the susceptible range to the highly resistant range, and as high as 37.1% fell into the high resistance of > 100 micrograms/ml range. 4. Against Peptostreptococcus spp. and MRSA, 16-membered macrolides were more effective than 14-membered macrorlides, and their antibacterial activities were in the order of RKM, JM, MDM. Ratio of high-resistant strains of > 100 micrograms/ml against 14-membered macrolides was much higher than that against 16-membered macrolies. 5. Most of M. (B.) catarrhalis strains were distributed in the susceptible range of < or = 1.56 micrograms/ml, and most of H. influenzae strains were distributed within the moderately resistant and the resistant ranges. 6. In M. (B.) catarrhalis and H. influenzae, no correlation between macrolide resistance and beta-lactamase production was recognized. 7. Most of C. jejuni strains were susceptible to all macrolides used in this study.     

Human complement component C1q inhibits the infectivity of cell-free HTLV-I

Human T cell leukemia virus type I (HTLV-I) is a retrovirus that is not lysed by human serum or complement. It has not been determined, however, whether HTLV-I directly binds to complement components or whether it retains infectivity after incubation with human serum. We investigated the effects of human serum on the infectivity of cell-free HTLV-I produced by human and animal cells. Plating of vesicular stomatitis virus (HTLV-I) pseudotypes prepared in cat or human cells and formation of HTLV-I DNA after infection of cell-free HTLV-I produced by cat or human cells were markedly inhibited by treatment with fresh human serum, but not by heat-inactivated serum. HTLV-I infection was also inhibited by treatment with C2-, C3-, C6-, or C9-deficient serum, but not by C1q-deficient serum. Inhibitory activities of normal human serum against HTLV-I were neutralized by anti-C1q serum. Furthermore, purified C1q inhibited HTLV-I infection. The direct binding of C1q to HTLV-I was confirmed by comigration of C1q with HTLV-I virion upon sucrose density gradient ultracentrifugation of HTLV-I virion treated with C1q. Binding assay using synthetic envelope peptides indicated that C1q bound to an extramembrane region of the gp21 transmembrane protein. These findings indicate that the human complement component C1q inactivates HTLV-I infectivity.     

Leaching and cytotoxicity of formaldehyde and methyl methacrylate from acrylic resin denture base materials

Acrylic resin dentures have the potential to elicit irritation, inflammation, and an allergic response of the oral mucosa. Studies of substances leachable from acrylic resins, their cytotoxicity to cultured cells, and means of reducing their leaching were systematically conducted. Under in vivo and in vitro conditions, formaldehyde and methyl methacrylate were significantly leached into human saliva and saliva-substitute buffer, especially from autopolymerized resins. Both leachable substances showed cytotoxic potentials in the range of their leaching concentrations. Formaldehyde was cytotoxic at lower concentrations than methyl methacrylate. Preleaching in water reduced subsequent leaching of both formaldehyde and methyl methacrylate, and the amount of reduction depended on an increase in the preleaching temperatures. Immersion of acrylic resin dentures in hot water (50 degrees C) before insertion is recommended, especially for autopolymerized resins used either for rebasing or as denture base materials, to minimize the risk of adverse reactions in patients who wear acrylic resin dentures.