Change of pupil size as a function of exercise.

Exercise was found to dilate the pupil area while the exercise took place, while the area showed constriction following the exercise period. Exercise-induced change in the size of the pupil was minimal. Pupillary dilation was greatest under conditions of maximal exercise. In exercise under a consistent load, pupillary dilation increased as the exercise time was prolonged. With lower lighting, there was virtually no dilation with exercise.     

Molecular spectrum of somaclonal variation in regenerated rice revealed by whole-genome sequencing

Somaclonal variation is a phenomenon that results in the phenotypic variation of plants regenerated from cell culture. One of the causes of somaclonal variation in rice is the transposition of retrotransposons. However, many aspects of the mechanisms that result in somaclonal variation remain undefined. To detect genome-wide changes in regenerated rice, we analyzed the whole-genome sequences of three plants independently regenerated from cultured cells originating from a single seed stock. Many single-nucleotide polymorphisms (SNPs) and insertions and deletions (indels) were detected in the genomes of the regenerated plants. The transposition of only Tos17 among 43 transposons examined was detected in the regenerated plants. Therefore, the SNPs and indels contribute to the somaclonal variation in regenerated rice in addition to the transposition of Tos17. The observed molecular spectrum was similar to that of the spontaneous mutations in Arabidopsis thaliana. However, the base change ratio was estimated to be 1.74 × 10(-6) base substitutions per site per regeneration, which is 248-fold greater than the spontaneous mutation rate of A. thaliana.     

Various adhesion molecules impair microvascular leukocyte kinetics in ventilator-induced lung injury

Although the endothelial expression of various adhesion molecules substantially differs between pulmonary microvessels, their importance for neutrophil and lymphocyte sequestration in ventilator-induced lung injury (VILI) has not been systematically analyzed. We investigated the kinetics of polymorphonuclear cells (PMN) and mononuclear cells (MN) in the acinar microcirculation of the isolated rat lung with VILI by real-time confocal laser fluorescence microscopy, with or without inhibition of ICAM-1, VCAM-1, or P-selectin by monoclonal antibodies (MAb). Adhesion molecules in each microvessel were estimated by intravital fluorescence microscopy or immunohistochemical staining. In high tidal volume-ventilated lungs, 1) ICAM-1, VCAM-1, and P-selectin were differently upregulated in venules, arterioles, and capillaries; 2) venular PMN rolling was improved by inhibition of ICAM-1, VCAM-1, or P-selectin, whereas arteriolar PMN rolling was improved by ICAM-1 or VCAM-1 inhibition; 3) capillary PMN entrapment was ameliorated only by anti-ICAM-1 MAb; and 4) MN rolling in venules and arterioles and MN entrapment in capillaries were improved by ICAM-1 and VCAM-1 inhibition. In conclusion, the contribution of endothelial adhesion molecules to abnormal leukocyte behavior in VILI-injured microcirculation is microvessel and leukocyte specific. ICAM-1- and VCAM-1-dependent, but P-selectin-independent, arteriolar PMN rolling, which is expected to reflect the initial stage of tissue injury, should be taken as a phenomenon unique to ventilator-associated lung injury.     

Identification of a putative voltage-gated Ca2+ -permeable channel (OsTPC1) involved in Ca2+ influx and regulation of growth and development in rice

Cytosolic free Ca2+ serves as an important second messenger participating in signal transduction of various environmental stresses. However, molecular bases for the plasma membrane Ca2+ influx and its regulation remain largely unknown. We here identified a gene (OsTPC1) encoding a putative voltage-gated Ca2+ channel from rice, ubiquitously expressed in mature leaves, shoots and roots as well as in cultured cells. OsTPC1 rescued the Ca2+ uptake activity and growth rate of a yeast mutant cch1. To elucidate its physiological roles, we generated transgenic rice plants and cultured cells overexpressing OsTPC1 mRNA. Furthermore, a retrotransposon (Tos17) insertional knockout mutant of OsTPC1 was isolated. OsTPC1-overexpressing cells showed hypersensitivity to excess Ca2+ but higher growth rate under Ca2+ limitation, while growth of the OsTPC1-knockout cultured cells was less sensitive to extracellular free Ca2+ concentration, suggesting that OsTPC1 has Ca2+ transport activity across the plasma membrane. OsTPC1-overexpressing plants showed reduced growth and abnormal greening of roots. Growth of Ostpc1 seedlings was comparable to the control on agar plates, while significantly reduced in adult plants. These results suggest that OsTPC1 functions as a Ca2+ -permeable channel involved in the regulation of growth and development.     

Treatment of glioblastoma by direct inoculation of concentrated high titer-recombinant retrovirus carrying the herpes simplex virus thymidine kinase gene.

We prepared retroviruses carrying the lacZ gene or herpes simplex virus thymidine kinase (HTK) gene with titers of 1.4-2.5 x 10(11) colony-forming units (cfu)/ml, and stereotaxically inoculated only 3 microliters of the retroviruses into a mouse glioma model. This resulted in highly efficient transduction in vivo. The transduced glioma cells migrated far from the implantation site, potentiating the induction of the remarkable bystander effect. Following repetitive ganciclovir (GCV) intraperitoneal injection, effective killing of glioma cells in the mouse brain was observed. The transduction efficiency was nearly as high as that observed for the implantation of high-titer retrovirus-producing fibroblasts. Eighty per cent of brain tumor-bearing mice were completely cured by our treatment protocol using concentrated HTK-harboring retroviruses. Our results suggest that repeated inoculations of high-titer retroviruses carrying the HTK gene followed by GCV treatment may be a promising strategy for the clinical treatment of malignant gliomas. To achieve further safety in the gene therapy of glioma, genes abundantly expressed in human glioblastoma were searched by the Serial Analysis of Gene Expression (SAGE) technique. Among the top-147 most expressed tags in glioblastoma, we found a tag, TTTTGGGTAT, originated from an unidentified gene, which was not detected in human astrocyte cultures. Real-time quantitative RT-PCR showed that MAGE-E1 expression was 2.6-15 fold enriched in glioblastoma relative to human astrocytes. Expressed Sequence Tags (ESTs) containing this tag were homologous to melanoma-associated antigen gene (MAGE) family, and this new cDNA, named MAGE-E1, was cloned by 5'-rapid amplification of cDNA ends (RACE) technique. MAGE-E1 expression was enriched in glioblastoma and low in other cancers, and MAGE-E1 expression was detected only in brain and ovary among normal tissues. These results indicate that MAGE-E1 is a novel and glioma-specific member of MAGE family, which can be applied to glioma-specific gene transduction.     

Precipitating antibody against protease of Corynebacterium pyogenes in pigs.

Antibody in sera from pigs carrying an abscess associated with Corynebacterium pyogenes and healthy pigs was examined by the agar gel diffusion test. In the test, the concentrated culture fluid containing the protease of C. pyogenes was used as antigen. As a result, precipitating antibody was demonstrated in sera from 25 of 30 abscessed pigs and a few of the healthy pigs. When the relationship between precipitating antibody and protease was examined by the immunoelectrophoresis and gel filtration of the concentrated culture fluid, the antibody was shown in the same position as the protease. From the result, it was clear that the precipitating antibody was against the protease of C. pyogenes. All the proteases produced by 27 strains of C. pyogenes of porcine and bovine origin were serologically identical with one another. They were, however, serologically different from those of Staphylococcus aureus, Bacillus cereus, and B. subtilis. In the inhibition test, the proteolytic activity of C. pyogenes was inhibited by the serum of the abscessed pig. It was also inhibited by healthy pig serum. From the results, it seems that the determination of precipitating antibody may be useful for the diagnosis of C. pyogenes infection.     

Physiological and genetic analyses of aluminium tolerance in rice, focusing on root growth during germination

Aluminium (Al) ion limits root growth of plants in acidic soils, and rice exhibits the highest level of Al-tolerance among graminous crops. To elucidate Al-tolerance mechanisms in rice, response to Al was compared between rice (Oryza sativa L., cv. Nipponbare) and wheat (Triticum aestivum L., cv. ET8), focusing on seminal root growth at seedling stage and germination stage. At seedling stage, rice and wheat were similarly sensitive to Al in both dose- and time-dependent manner during a 24-h Al exposure. On the contrary, at germination stage, rice was more tolerant to Al than wheat, and wheat roots displayed the loss of plasma membrane integrity more extensively than rice. A rice mutant exhibiting Al hypersensitivity at germination stage was obtained by screening 42,840 R2 progeny derived from the regenerated R0 plants of Nipponbare and thereafter confirmation of the mutant phenotype in R3 progeny. At germination stage, root growth of the mutant was strongly inhibited in the presence of Al but not in the absence of Al. However, at seedling stage, root growth of the mutant and wild type was similarly tolerant to Al. Taken together, we conclude that rice possesses Al-tolerant function that is under genetic control and specifically operates for root growth at germination stage, making rice more tolerant to Al than wheat.