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.     

Cloning and characterization of elastase structural gene from Pseudomonas aeruginosa IFO 3455

An 8.3 Kb DNA fragment was cloned from Pseudomonas aeruginosa IFO 3455. This fragment-containing Escherichia clone, pEL2, produced a high level of elastase activity. A smaller EcoRI-KpnI fragment was subcloned into pUC118 and E. coli HB101 was transformed with the plasmid. A deletion mutant clone was also constructed in the same bacteria. These deletion mutants were tested for elastase activity and it became clear that the full length of the elastase gene was 1.0-1.3 Kb. DNA sequencing analysis revealed that this DNA fragment contains the DNA sequence coding N-terminal amino acid sequence of the elastase protein.     

Adsorption of mutagens by refined corn bran

Refined corn bran (RCB), a dietary fiber derived from the mechanical refining of corn hulls, effectively adsorbed various environmental mutagens. When RCB was added at a concentration of 10 mg/ml to an aqueous solution of dinitropyrene (DNP), 91.6% of the mutagenicity towards Salmonella tester strain TA98 disappeared. Under similar conditions decreases in mutagenicity of DNP using wheat bran and cellulose powder were 58.4% and 43.0%, respectively. The adsorption of DNP to the fibers appeared irreversible since little mutagenicity was recovered by washing the treated fibers with aqueous buffer solutions of various pHs. Even with an organic solvent (methanol: ammonium hydroxide 50:1), only 2/3 of the mutagenicity of DNP was recovered. RCB could similarly adsorb mutagenic heterocyclic amines such as IQ, Trp-P-1, Trp-P-2, Glu-P-1, and Glu-P-2.     

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.     

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     

Elementary steps in the acto-H-meromyosin ATPase reaction to arterial smooth muscle.

Transient and steady state kinetics were studied in the interactions of ATP with acto-H-meromyosin reconstituted from bovine arterial heavy-meromyosin (HMM) and rabbit skeletal muscle F-actin. The results showed that the rate of dissociation of the hybrid acto-HMM induced by ATP was slower than the rate of the fluorescence enhancement of HMM, and that the rate of the P1 burst of HMM was unaffected by addition of skeletal muscle F-actin. The ATPase [EC 3.6.1.3] activity of arterial HMM was activated only slightly even with addition of high concentrations of skeletal muscle F-actin. Furthermore, the rates of dissociation of the hybrid acto-HMM induced by ATP and reassociation of dissociated arterial HMM with skeletal muscle F-actin after decomposition of ATP were much lower than those of skeletal muscle acto-HMM.     

Behavior of a vigorous alpha- or beta-hemolysin-producing strain of Staphylococcus aureus in the cutaneous tissue of mice.

Two strains of Staphylococcus aureus were examined for behavior in the cutaneous tissue of mice by the fluorescent antibody technique, hematoxylin and eosin staining. When about 10(8) viable cells of an alpha-hemolysin-producing strain (Wood 46) were inoculated subcutaneously into a mouse, they multiplied in the subcutaneous tissue of the mouse and gradually entered the corium to produce alpha-hemolysin and nuclease. Edematous and necrotic lesions were observed in the cutaneous tissue where the organisms had multiplied. When 10(8) viable cells of a beta-hemolysin-producing strain (Kitami 3-9D) were inoculated into a mouse, they multiplied within a narrow extent surrounded mainly by infiltrating leukocytes and produced mainly beta-hemolysin. The changes of cutaneous tissue were weaker in mice inoculated with Kitami 3-9D strain than in mice inoculated with strain Wood 46. When 10(6) viable cells of both strains were inoculated into mice, they were phagocytized by leukocytes. Neither multiplication of organisms nor production of any active extracellular substance was observed in these mice. Edema, degeneration, and necrosis were also noticed in the cutaneous tissue of mice inoculated with alpha- and beta-hemolysin. In addition, the infiltration of leukocytes was inhibited mainly by alpha-hemolysin.