Decontamination of mycoplasma-contaminated Orientia tsutsugamushi strains by repeating passages through cell cultures with antibiotics

Background: Mycoplasmas-contamination of Orientia tsutsugamushi, one of the obligated intracellular bacteria, is a very serious problem in in vitro studies using cell cultures because mycoplasmas have significant influence on the results of scientific studies. Only a recommended decontamination method is to passage the contaminated O. tsutsugamushi strains through mice to eliminate only mycoplasmas under influence of their immunity. However, this method sometimes does not work especially for low virulent strains of O. tsutsugamushi which are difficult to propagate in mice. In this study, we tried to eliminate mycoplasmas contaminants from both high virulent and low virulent strains of the contaminated O. tsutsugamushi by repeating passage through cell cultures with antibiotics in vitro. Results: We cultured a contaminated, high virulent strain of O. tsutsugamushi using a mouse lung fibroblasts cell line, L-929 cell in the culture medium containing lincomycin at various concentrations and repeated passages about every seven days. At the passage 5 only with 10 μg/ml of lincomycin, we did not detect mycoplasmas by two PCR based methods whereas O. tsutsugamushi continued good growth. During following four passages without lincomycin, mycoplasmas did not recover. These results suggested that mycoplasmas were completely eliminated from the high virulent strain of O. tsutsugamushi. Furthermore, by the same procedures with 10 μg/ml of lincomycin, we also eliminated mycoplasmas from a contaminated, low virulent strain of O. tsutsugamushi. Our additional assay showed that 50 μg/ml of lyncomycin did not inhibit the growth of O. tsutsugamushi, although MICs of many mycoplasmas contaminants were less than 6 μg/ml as shown previously. Conclusion: Our results showed an alternative method to eliminate mycoplasmas from the contaminated O. tsutsugamushi strains in place of in vivo passage through mice. Especially this notable method works for the decontamination not only from the high virulent strain also from the low virulent strain of O. tsutsugamushi. For further elimination, lincomycin at the limit concentration, which does not inhibit the growth of O. tsutsugamushi, can possibly eliminate most mycoplasmas from contaminated O. tsutsugamushi strains

Statistical study of Doppler velocity and echo power around 75゜ magnetic latitude using data obtained with the Syowa East HF radar in 1997

We present the statistical relationship between the echo power and Doppler velocity of radar echoes observed with the Syowa East HF radar in Antarctica in 1997. The objective of this analysis was to clarify the mechanisms by which high-latitude (～75° magnetic latitude) F region irregularities are generated. Although data points are scattered over a large area, a positive correlation between Doppler velocity and echo power appears to be present. This relationship can be interpreted in terms of gradient-drift instability, which is the most probable cause of the decameter-scale irregularities in the F region. The positive correlation deteriorates in the afternoon and midnight sectors, probably as a result of other mechanisms related to particle precipitation (field-aligned current), such as the current-convective instability

Statistical analysis of echo power, Doppler velocity and spectral width obtained with the Syowa South HF radar

Statistical analyses are made of the physical parameters (echo power, Doppler velocity and spectral width) of Doppler spectra obtained in September 1995 with the Syowa South HF radar. We present time and range distributions and histograms of the parameters and cross-correlations among them. With K index at Syowa Station the distributions and histograms vary slightly but the correlations do not change so much. The most noticeable feature is that there is a positive correlation between the absolute values of Doppler velocity and echo power. This relationship can be interpreted in terms of the gradient-drift instability which is the most probable cause to generate decameter-scale irregularities in the F-region ionosphere

Potent Inhibitory Effects of D-tagatose on the Acid Production and Water-insoluble Glucan Synthesis of Streptococcus mutans GS5 in the Presence of Sucrose

We examined and compared the inhibitory effects of D-tagatose on the growth, acid production, and water-insoluble glucan synthesis of GS5, a bacterial strain of Streptococcus mutans, with those of xylitol, D-psicose, L-psicose and L-tagatose. GS5 was cultured for 12h in a medium containing 10ｵ (w/v) of xylitol, D-psicose, L-psicose, D-tagatose or L-tagatose, and the inhibitory effect of GS5 growth was assessed. Each sugar showed different inhibitory effects on GS5. Both D-tagatose and xylitol significantly inhibited the acid production and water-insoluble glucan synthesis of GS5 in the presence of 1ｵ (w/v) sucrose. However, the inhibitory effect of acid production by D-tagatose was significantly stronger than that of xylitol in presence of sucrose

Aur-A Stabilization in Cancer

Background. The serine/threonine kinase Aurora-A (Aur-A) is a proto-oncoprotein overexpressed in a wide range of human cancers. Overexpression of Aur-A is thought to be caused by gene amplification or mRNA overexpression. However, recent evidence revealed that the discrepancies between amplification of Aur-A and overexpression rates of Aur-A mRNA were observed in breast cancer, gastric cancer, hepatocellular carcinoma, and ovarian cancer. We found that aggressive head and neck cancers exhibited overexpression and stabilization of Aur-A protein without gene amplification or mRNA overexpression. Here we tested the hypothesis that aberration of the protein destruction system induces accumulation and consequently overexpression of Aur-A in cancer. Principal Findings. Aur-A protein was ubiquitinylated by APCCdh1 and consequently degraded when cells exited mitosis, and phosphorylation of Aur-A on Ser51 was observed during mitosis. Phosphorylation of Aur-A on Ser51 inhibited its APCCdh1-mediated ubiquitylation and consequent degradation. Interestingly, constitutive phosphorylation on Ser51 was observed in head and neck cancer cells with protein overexpression and stabilization. Indeed, phosphorylation on Ser51 was observed in head and neck cancer tissues with Aur-A protein overexpression. Moreover, an Aur-A Ser51 phospho-mimetic mutant displayed stabilization of protein during cell cycle progression and enhanced ability to cell transformation. Conclusions/Significance. Broadly, this study identifies a new mode of Aur-A overexpression in cancer through phosphorylation-dependent inhibition of its proteolysis in addition to gene amplification and mRNA overexpression. We suggest that the inhibition of Aur-A phosphorylation can represent a novel way to decrease Aur-A levels in cancer therapy

Neural Correlates of Stimulus–Response and Response–Outcome Associations in Dorsolateral Versus Dorsomedial Striatum

Considerable evidence suggests that there is functional heterogeneity in the control of behavior by the dorsal striatum. Dorsomedial striatum may support goal-directed behavior by representing associations between responses and outcomes (R–O associations). The dorsolateral striatum, in contrast, may support motor habits by encoding associations between stimuli and responses (S–R associations). To test whether neural correlates in striatum in fact conform to this pattern, we recorded single-units in dorsomedial and dorsolateral striatum of rats performing a task in which R–O contingencies were manipulated independently of S–R contingencies. Among response-selective neurons in both regions, activity was significantly modulated by the initial stimulus, providing evidence of S–R encoding. Similarly, response selectivity was significantly modulated by the associated outcome in both regions, providing evidence of R–O encoding. In both regions, this outcome-modulation did not seem to reflect the relative value of the expected outcome, but rather its specific identity. Finally, in both regions we found correlates of the available action–outcome contingencies reflected in the baseline activity of many neurons. These results suggest that differences in information content in these two regions may not determine the differential roles they play in controlling behavior, demonstrated in previous studies

Subcutaneous Single Injection Digital Block with Epinephrine

The aim of this study was to investigate the anesthetic effect and risk of epinephrine for subcutaneous single injection digital block. Either 3.0 mL 1.0% Lidocaine or a 3.0 mL 1.0% Lidocaine with (1 : 100,000) epinephrine was injected into the subcutaneous space at the middle point of the palmar digital crease of the 18 middle fingers of 9 healthy volunteers. The SpO2 of the fingers decreased to a maximum of 97. No subjects showed any symptoms of ischemic injury. The time to anesthesia for the fingers was significantly shorter (P < 0.05), and the duration of anesthesia was significantly longer (P < 0.01) for the fingers in the epinephrine group. In conclusion, a subcutaneous single injection digital blocks with 3.0 mL of 1.0% Lidocaine and (1 : 100,000) epinephrine were safe, reducing the time to the onset of anesthesia, while also markedly prolonging the anesthesia