Studies on live rubella vaccine. VI. Seroepidemiological surveillance and immunization of adolescents in high school.

The rubella seroimmunity status of a total of 1,204 students aged 12 to 19 in a junior and a senior high school in Osaka district was surveyed. Among these, 487 students (40.5%) were found to be seronegative (less than 1 : 8) by the hemagglutination inhibition (HI) test. A total of 287 students were immunized with live rubella vaccine, Biken lot No.7233. This caused an increased titer in all except one of the 262 seronegative students, while among 25 students with an initial HI antibody titer of 1 : 8 it caused more than 4-fold increase in 20 and 2-fold increase in 5. The vaccine caused no clinical manifestations, such as fever, rash or arthralgia.     

Productivity of immunoreactive prolactin (IR-PRL) by the human decidual explants: a new sampling method

In the present study, we used an explant culture system of the human decidual tissues involving a new sampling method for investigating the productivity of immunoreactive prolactin (IR-PRL) for a 5 day period in labored and nonlabored deliveries. The maximal release of IR-PRL in the incubation medium for each 24 hour interval was achieved from the 2nd to the 3rd day of culture in both groups, which was 145.2 +/- 14.0 ng/ml/10 mg w.w. (mean +/- s.e.; n = 7) in the labor group and 101.5 +/- 16.3 ng/ml/10 mg w.w. (mean +/- s.e.; n = 4) in the nonlabor group. The release of IR-PRL in both groups was not significantly different for the first 3 days. However, the amount of IR-PRL released in the nonlabor group was 50 to 70% of that of the labor group. The tissue content of IR-PRL in both groups ranged between 3.0 and 5.0 ng/ml/10 mg w.w. From these results, it was concluded that 1) our explant culture system for the human decidual tissues produced considerably more IR-PRL than those previously reported and 2) productivity of IR-PRL was lower in nonlabored delivery than in labored delivery and 3) since the tissue content of IR-PRL for each 24 hour interval was very small, it should be strongly emphasized that the production and release of IR-PRL takes place simultaneously in the human decidual tissues.     

Visualization of Cortical Projection Neurons with Retrograde TET-Off Lentiviral Vector

We are interested in identifying and characterizing various projection neurons that constitute the neocortical circuit. For this purpose, we developed a novel lentiviral vector that carries the tetracycline transactivator (tTA) and the transgene under the TET Responsive Element promoter (TRE) on a single backbone. By pseudotyping such a vector with modified rabies G-protein, we were able to express palmitoylated-GFP (palGFP) or turboFP635 (RFP) in corticothalamic, corticocortical, and corticopontine neurons of mice. The high-level expression of the transgene achieved by the TET-Off system enabled us to observe characteristic elaboration of neuronal processes for each cell type. At higher magnification, we were able to observe fine structures such as boutons and spines as well. We also injected our retrograde TET-Off vector to the marmoset cortex and proved that it can be used to label the long-distance cortical connectivity of millimeter scale. In conclusion, our novel retrograde tracer provides an attractive option to investigate the morphologies of identified cortical projection neurons of various species.     

The Role of Catalase in Pulmonary Fibrosis

Background

Catalase is preferentially expressed in bronchiolar and alveolar epithelial cells, and acts as an endogenous antioxidant enzyme in normal lungs. We thus postulated epithelial damage would be associated with a functional deficiency of catalase during the development of lung fibrosis.

Methods

The present study evaluates the expression of catalase mRNA and protein in human interstitial pneumonias and in mouse bleomycin-induced lung injury. We examined the degree of bleomycin-induced inflammation and fibrosis in the mice with lowered catalase activity.

Results

In humans, catalase was decreased at the levels of activity, protein content and mRNA expression in fibrotic lungs (n = 12) compared to control lungs (n = 10). Immunohistochemistry revealed a decrease in catalase in bronchiolar epithelium and abnormal re-epithelialization in fibrotic areas. In C57BL/6J mice, catalase activity was suppressed along with downregulation of catalase mRNA in whole lung homogenates after bleomycin administration. In acatalasemic mice, neutrophilic inflammation was prolonged until 14 days, and there was a higher degree of lung fibrosis in association with a higher level of transforming growth factor-β expression and total collagen content following bleomycin treatment compared to wild-type mice.

Conclusions

Taken together, these findings demonstrate diminished catalase expression and activity in human pulmonary fibrosis and suggest the protective role of catalase against bleomycin-induced inflammation and subsequent fibrosis.     

Design, syntheses, and structure-activity relationships of novel NPY Y5 receptor antagonists: 2-{3-Oxospiro[isobenzofuran-1(3H),4'-piperidin]-1'-yl}benzimidazole derivatives

Design, syntheses, and structure-activity relationships of a novel class of 2-{3-oxospiro[isobenzofuran-1(3H),4'-piperidin]-1'-yl}benzimidazole NPY Y5 receptor antagonists are described. The benzimidazole structures were newly designed based on the urea linkage of our prototype Y5 receptor antagonists (2 and 3). By optimizing substituents on the benzimidazole core part of the lead compound 5a, we were able to develop a potent, orally available, and brain-penetrable Y5 selective antagonist (5k).     

Enhancement of the organic solvent‐stability of the LST‐03 lipase by directed evolution

LST‐03 lipase from an organic solvent‐tolerant Pseudomonas aeruginosa LST‐03 has high stability and activity in the presence of various organic solvents. In this research, enhancement of organic solvent‐stability of LST‐03 lipase was attempted by directed evolution. The structural gene of the LST‐03 lipase was amplified by the error prone‐PCR method. Organic solvent‐stability of the mutated lipases was assayed by formation of a clear zone of agar which contained dimethyl sulfoxide (DMSO) and tri‐n‐butyrin and which overlaid a plate medium. And the organic solvent‐stability was also confirmed by measuring the half‐life of activity in the presence of DMSO. Four mutated enzymes were selected on the basis of their high organic solvent‐stability in the presence of DMSO. The organic solvent‐stabilities of mutated LST‐03 lipase in the presence of various organic solvents were measured and their mutated amino acid residues were identified. The half‐lives of the LST‐03‐R65 lipase in the presence of cyclohexane and n‐decane were about 9 to 11‐fold longer than those of the wild‐type lipase, respectively. Some substituted amino acid residues of mutated LST‐03 lipases have been located at the surface of the enzyme molecules, while some other amino acid residues have been changed from neutral to basic residues. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009