Nitrile hydratase from a thermophilic Bacillus smithii

A novel thermophilic Bacillus smithii strain SC-J05-1, isolated from a hot spring, had the ability of hydrating nitrile to form amide. The nitrile hydratase was purified to homogeneity from the microbial cells of SC-J05-1 and was characterized. The enzyme was a 130-kDa protein composed of two different subunits (25.3 kDa and 26.8 kDa) and contained cobalt ions. This enzyme had the optimal temperature of 40°C and was stable up to 50°C. The optimal pH was in the alkaline region higher than pH 10. Received 02 September 1997/ Accepted in revised form 06 February 1998     

Cloning and antigenic expression of two genes from Chlamydia psittaci avian strain

Abstract: Genes from Chlamydia psittaci P-1041 were cloned into the Bam HI site of pUC19 and were transformed to host Escherichia coli JM109. Two recombinant plasmids that expressed protein antigens of Chlamydia were isolated. The sizes of the DNA fragments were 1350 and 1710 bp, and encoded for polypeptides of M _r 25 and 42 kilodaltons (kDa), respectively. The 25-kDa protein had cross-reactivity with antisera to ten C. psittaci strains and two C. trachomatis strains, whereas the 42-kDa protein reacted only with homologous antiserum to the C. psittaci P-1041 strain. Furthermore, in Southern hybridization analysis these two fragments as probes hybridized with DNA of ten C. psittaci strains and four C. trachomatis strains. These results indicated that the two fragments shared a DNA sequence common to the chlamydial genus.     

Essential Role of the m2R-RGS6-IKACh Pathway in Controlling Intrinsic Heart Rate Variability

Normal heart function requires generation of a regular rhythm by sinoatrial pacemaker cells and the alteration of this spontaneous heart rate by the autonomic input to match physiological demand. However, the molecular mechanisms that ensure consistent periodicity of cardiac contractions and fine tuning of this process by autonomic system are not completely understood.Here we examined the contribution of the m₂R-I_KACh intracellular signaling pathway, which mediates the negative chronotropic effect of parasympathetic stimulation, to the regulation of the cardiac pacemaking rhythm. Using isolated heart preparations and single-cell recordings we show that the m₂R-I_KACh signaling pathway controls the excitability and firing pattern of the sinoatrial cardiomyocytes and determines variability of cardiac rhythm in a manner independent from the autonomic input. Ablation of the major regulator of this pathway, Rgs6, in mice results in irregular cardiac rhythmicity and increases susceptibility to atrial fibrillation. We further identify several human subjects with variants in the RGS6 gene and show that the loss of function in RGS6 correlates with increased heart rate variability. These findings identify the essential role of the m₂R-I_KACh signaling pathway in the regulation of cardiac sinus rhythm and implicate RGS6 in arrhythmia pathogenesis.     

Development of the hand and wrist bones in chimpanzees

Skeletal developmental of chimpanzees was studied cross-sectionally. By application of the TW2 method, we described the skeletal development of chimpanzees and compared their skeletal development with humans'. A development pattern of chimpanzees repeated accelerations and decelerations displaying “early-juvenile trough,” “pre-adolescent peak,” “mid-adolescent trough,” and “post-adolescent peak” in incremental curves. Sex differences in skeletal development are slower development in males during infant and early juvenile phases, and greater increment around the adolescent phase in males. Females are fully mature at younger ages than males, e.g. about one and a half years. In comparison with chimpanzees, humans have such characteristics as a longer slower period of juvenile development and a shorter spurt-like adolescent fast period which ends with full maturity.     

Expression level of enzymes related to in situ estrogen synthesis and clinicopathological parameters in breast cancer patients

In order to evaluate the importance of estrogen production in tumor and surrounding tissues, we measured mRNA expression levels of 5 enzymes participating to estrogen synthesis in situ and 4 breast cancer-related proteins in 27 pairs of tumor and non-malignant tissues. Steroid sulfatase (STS) mRNA was more frequently detected in tumor tissues rather than in their non-malignant counterparts. Estrogen sulfotransferase (EST) was constantly expressed with high level not only in tumor tissues but also in their surrounding non-malignant counterparts. In contrast, mRNA expression levels of aromatase, and 17β-hydroxysteroid dehydrogenase type I and II were relatively low and detected only in small proportion of the patients. We also measured the mRNA expression levels of the same nine genes in tumor tissues of 197 breast cancer patients, and analyzed relationship between the mRNA expression level and the clinicopathological parameters. The mRNA expression levels of STS, aromatase and erbB2 in tumor tissues increased as breast cancer progressed. The tumoral mRNA expression levels of STS, estrogen receptor β, and erbB2 in patients with recurrence were higher than those in patients without recurrence. Upregulation of STS expression plays an important role in tumor progression of human breast cancer and is considered to be responsible for estrogen production in tumor and surrounding tissues.     

Echolocation sounds and hearing of the greater Japanese horseshoe bat (Rhinolophus ferrumequinum nippon)

Summary The relationship between the orientation sounds and hearing sensitivity in the greater Japanese horseshoe bat,Rhinolophus ferrumequinum nippon was studied.An orientation pulse consisted of a constant frequency (CF) component followed by a short downward frequency-modulated (FM) component. Sometimes, an initial upward FM component preceded the CF component. Duration of pulses was about 30 ms and the CF of resting pulses (RF) averaged 65.5 kHz. The best frequency (BF) at the lowest threshold in audiograms as measured by the pinna reflex averaged 66.1 kHz. Audiograms showed remarkable sharp cut-offs on both sides near the BF. The frequency difference between the BF and the RF was about 0.6 kHz, and the RF was always below the BF. The values of RF and BF were characteristically different from those of the European subspecies,Rhinolophus ferrumequinum ferrumequinum.Abbreviations BF best frequency - CF constant frequency - FM frequency modulated - RF resting frequency     

Non-empirical valence bond calculation of hydrogen bond energy in polypeptides

An approximate ab-initio valence bond calculation of hydrogen bond energy was carried out and the results are discussed. The total bond energy of a simplified N? H…O structure is calculated for various N? H and N…O distances and the potential energy profiles are obtained. The hydrogen bond energy, ie, the delocalization energy gained by the formation of one hydrogen bond is found to be 8.7–12.0 kcal/mole. The potential energy is characterized by a deep minimum at 1.6–1.8 a.u. from the nitrogen and the second trough is found to be considerably higher than the first.     

De novo glucan synthesis by mutans streptococcal glucosyltransferases present in pellicle promotes firm binding of Streptococcus gordonii to tooth surfaces

Adherence of 3H-labelled cells of Streptococcus gordonii and Streptococcus milleri to artificial pellicles prepared from saliva supplemented with glucosyltransferases from mutants streptococci was examined using a new assay for sucrose-dependent cell-to-pellicle attachment. Results indicate that S. gordonii, but not S. milleri, could attach tightly to hydroxylapatite surfaces through de novo glucan synthesis by mutants streptococcal glucosyltransferases present in the experimental salivary pellicles.