Focus Group Abstracts

Sleep and Biological Rhythms -     

DNA rearrangement associated with the integration of T-DNA in tobacco: an example for multiple duplications of DNA around the integration target

Transferred DNA (T-DNA) of the tumor-inducing (Ti) plasmid is transferred from Agrobacterium tumefaciens to plant cells and is stably integrated into the plant nuclear genome. By the inverse polymerase chain reaction DNA fragments were amplified that contained the T-DNA/plant DNA junctions from the total DNA of a transgenic tobacco plant that had a single copy of the T-DNA in a repetitive region of its genome. A DNA fragment containing the target site was amplified from the total DNA of non-transformed tobacco by the polymerase chain reaction using high-stringency conditions. Comparison of the nucleotide sequence of the target site with those of the T-DNA/plant DNA junctions revealed that various duplications of short stretches of nucleotide sequences around the target and in the incoming T-DNA had accompanied the integration of the T-DNA. A deletion of 16 bp at the target site was also found and the target site was similar, in terms of nucleotide sequence, to regions around the breakpoints of the T-DNA. This finding provides a clear example of the occurrence of complex rearrangements during the integration of T-DNA.     

The effect of long wave ultraviolet radiation (UV-A) on the photosynthetic activity of natural population of freshwater phytoplankton

The effect of ultraviolet radiation on diel changes and depth profiles of phytoplankton photosynthesis was studied in four temperate freshwater lakes. Photosynthetic oxygen production was determined by incubating lake water in light and dark bottles under various weather conditions. Half the light bottles were wrapped with sheets of vinyl chloride film to exclude light with wavelengths shorter than 400 nm. The inhibition of photosynthesis due to UV-A (320–400 nm) was observed during most of the daytime and was very strong around noon on both sunny and cloudy days. On sunny days, when the surface waters of the highly eutrophic Lake Suwa and Senzoku Pond were dominated by denseMicrocystis populations, cumulative daily production at the surface, estimated from the incubation of bottles from which UV-A was excluded by the vinyl film, were about double the rates obtained from glass bottles in which UV-A was present. The UV-A inhibition was detected from the surface toca 20 cm depth in hypereutrophic lakes and at depths greater than 50 cm in mesotrophic lakes. Analysis of the photosynthesis-irradiance (P-I) relationship obtained in the present study shows β, a parameter that describes photo-inhibition, is higher in the presence of UV-A than in its absence. This indicates that UV-A is the major cause of photo-inhibition of phytoplankton photosynthesis.     

Effect of a single administration of testosterone on the immune response and lymphoid tissues in mice.

Female mice were given a single intraperitoneal injection of testosterone immediately after irradiation and marrow reconstitution. Thirty days later testosterone had no suppressive effect on the recovery of thymus and spleen weights. Testosterone had no effect on the graft-versus-host reaction. Testosterone had no influence on the survival of the skin homografts. However, the plaque-forming cell response to sheep erythrocytes in the spleen was dramatically suppressed by testosterone. Histological observations revealed marked inhibition of lymphoid regeneration selectively in the thymus-independent areas of the peripheral lymphoid tissues. These results suggest that testosterone would act mainly on the differentiation of stem cells toward the population of bone marrow-derived B lymphocytes. The immune response to sheep erythrocytes was restored completely 90 days after testosterone administration. Testosterone given to normal adult mice can also have suppressive activity on the immune system 30 days after a single intraperitoneal injection.     

Perithecial formation in Gelasinospora reticulispora III. Inhibitory effects of near-UV and blue light during the inductive dark period

Growing hyphae of Gelasinospora reticulispora required a continuousdark period prior to photoinduction of perithecia. The inductivedark period was interrupted by brief exposure of the hyphaeto white light so that the formation of perithecia no longertook place. Photosensitivity of the hyphae in terms of the light-breakeffect gradually changed during the inductive dark period. Sensitivityreached its maximum at the 18th hr of the dark period when anirradiation of 1?10⁵ ergs cm^–2 of near-UV light or 4?10⁴ergs cm^–2 of blue-light was sufficient for the light-break.Red and far-red light had no effect at all. The light-breakeffect was limited to the irradiated portion of the hyphae anddid not affect any unirradiated portions. Inhibitory effecton perithecial formation of continuous white light could betotally replaced for several days with intermittent irradiationof near-UV or blue light if given for 5 min every 4 hr. (Received December 18, 1973; )     

Perithecial formation in Gelasinospora reticulispora II. Promotive effects of near-ultraviolet and blue light after dark incubation

Hyphae of Gelasinospora reticulispora growing on corn meal agarat 25?C required light of form perithecia. This response tolight was highly correlated to the length of the preliminarydark period, i.e. the photoinduction of perithecia never occurredin cultures grown in the dark for periods of 27 hr or less,whereas hyphae became photosensitive if incubated for 30 hror longer in the dark. The perithecia became simultaneouslyvisible with the 30 to 48 hr dark-grown hyphae irrespectiveof the different dark periods, but time courses for cultureshaving 54 hr or longer of darkness were dependent upon the timewhen light was given. Light induced perithecia only in the dark-grown portions ofthe hyphae. The longer the dark period, the more the sensitivityto light increased when the most effective range of wavelenghtswas shifted from near-ultraviolet to blue. Light of green andlonger wavelenghts was not at all effective irrespective ofthe duration of darkness. The photoinduced stimulus was notmovable from the irradiated to the unirradiated portions. (Received August 3, 1973; )     

Effects of Storage Temperature and Wounding on Cytokinin Levels in Potato Tubers

The level of butanol-soluble cytokinin in potato tubers (Solanumtuberosum L.) decreased greatly after harvest of the tubers,remained low during rest-period and then began to increase priorto the end of the period. In warm-stored tubers (25?C) the levelrose faster than in cold-stored ones (4?C). On the other hand,the level of water-soluble cytokinin, which was high duringthe rest-period, decreased after the rest was broken. Wounding the tubers caused an increase in butanol-soluble cytokinin.The increase in cytokinin within the first 24 hr after woundingwas smaller in warm-stored tubers than in cold-stored ones andwas decreased with increasing storage time of the tubers. Thelevel of water-soluble cytokinin decreased by wounding. Theseresults suggest that water-soluble cytokinin is a storage formand that the level of butanol-soluble cytokinin is regulatedby an interconversion between the two. (Received February 6, 1982; Accepted May 13, 1982)     

LPA1/3 signaling mediates tumor lymphangiogenesis through promoting CRT expression in prostate cancer

Lysophosphatidic acid (LPA) is a bioactive lipid growth factor which is present in high levels in serum and platelets. LPA binds to its specific G-protein-coupled receptors, including LPA₁ to LPA₆, thereby regulating various physiological functions, including cancer growth, angiogenesis, and lymphangiogenesis. Our previous study showed that LPA promotes the expression of the lymphangiogenic factor vascular endothelial growth factor (VEGF)-C in prostate cancer (PCa) cells. Interestingly, LPA has been shown to regulate the expression of calreticulin (CRT), a multifunctional chaperone protein, but the roles of CRT in PCa progression remain unclear. Here we investigated the involvement of CRT in LPA-mediated VEGF-C expression and lymphangiogenesis in PCa. Knockdown of CRT significantly reduced LPA-induced VEGF-C expression in PC-3 cells. Moreover, LPA promoted CRT expression through LPA receptors LPA₁ and LPA₃, reactive oxygen species (ROS) production, and phosphorylation of eukaryotic translation initiation factor 2α (eIF2α). Tumor-xenografted mouse experiments further showed that CRT knockdown suppressed tumor growth and lymphangiogenesis. Notably, clinical evidence indicated that the LPA-producing enzyme autotaxin (ATX) is related to CRT and that CRT level is highly associated with lymphatic vessel density and VEGF-C expression. Interestingly, the pharmacological antagonist of LPA receptors significantly reduced the lymphatic vessel density in tumor and lymph node metastasis in tumor-bearing nude mice. Together, our results demonstrated that CRT is critical in PCa progression through the mediation of LPA-induced VEGF-C expression, implying that targeting the LPA signaling axis is a potential therapeutic strategy for PCa.