Spatial distribution and mortality process ofAnoplophora malasiaca (Coleoptera: Cerambycidae) eggs in citrus groves

The study was carried out in two (A and B) citrus groves to clarify the spatial distribution patterns of eggs and larvae, and to analyse the mortality process of eggs. From the analysis by using the mean density and the mean crowding, it was clarified that the distributions of eggs were contagious and that larvae were more contagiously distributed than eggs. The τ and z indices showed that the operation of egg mortality was inversely density-dependent in both groves, and that the degree of inverse density-dependence was greater in A than in B grove. The spatial correlations between the emergence holes and the eggs or larvae in each tree, which were analysed by using ω index, showed that the distributions were more overlapping between the emergence holes and the larvae than the eggs. As the result of dividing trees into several groups according to the number of emergence holes, it was clarified that the survival rates of eggs were positively correlated with the number of emergence holes. In conclusion, inversely density-dependent mortality process was considered to be caused by lower mortality rates of eggs in the trees with more emergence holes. Especially in A grove, because the trees with more emergence holes were larger in diameter and more egg oviposition, the inversely density-dependent mortality process was considered to be detected more conspicuously than in B grove.     

The augmentation of tumor-specific immunity using haptenic muramyl dipeptide (MDP) derivatives

Summary A new haptenic compound, a muramyl dipeptide (MDP) derivative (designated as L4-MDP-ONB) cross-reactive with Bacillus Calmette Guerin (BCG) was synthesized. The cross-reactivity of L4-MDP hapten to BCG was demonstrated from the following evidence; (a) lymph node cells from BCG-primed C3H/HeN mice exhibited appreciable L4-MDP-specific proliferative responses to the in vitro stimulation of L4-MDP-modified syngeneic cells (L4-MDP-self); (b) inoculation of L4-MDP-self into footpads of BCG-primed C3H/HeN mice elicited ample delayed type-hypersensitivity (DTH) responses in vivo as measured by footpad swelling; and (c) BCG-primed mice contained L4-MDP-reactive helper T cell activity which functions to augment the generation of effector T cell responses to cell surface antigens. This crossreactivity between L4-MDP hapten and BCG as measured by the helper T cell activity was applied to enhanced induction of tumor-specific immunity. When BCG-primed C3H/HeN mice were immunized with L4-MDP-modified syngeneic X5563 tumor cells, these mice could generate augmented tumor-specific in vivo protective (tumor neutralizing) immunity as well as in vitro cytotoxic T cell responses. These results indicate the effectiveness of L4-MDP hapten in augmenting tumor-specific immunity. The present approach is discussed in the context of potential advantages of this new hapten for its future application to clinical tumor systems.     

Aggregation of hemoglobin S modified by bifunctional imidoesters

Sickle hemoglobin (Hb S) was cross-linked by two types of bifunctional imidoesters, dimethyladipimidate (DMA) and dimethyl-3,3'-dithiobispropionimidate (DTBP). These modified hemoglobins were separated into monomer, dimer and polymer fractions by gel filtration. All of these modified hemoglobins showed extremely left-shifted oxygen equilibrium curves with no cooperativity. The stabilities of these hemoglobins were also decreased. The solubilities of these modified hemoglobins in high-phosphate buffers were lower than those of native Hb S. Studies on the kinetics of the aggregation of these modified hemoglobins showed that intracross-linked Hb S with DMA and DTBP (DMA- and DTBP-modified monomeric Hb S) still retained the capability of aggregation with a delay time, while intercross-linked Hb S with DMA and DTBP (DMA- and DTBP-modified oligomeric Hb S) aggregated without a delay time. When the kinetics of aggregation was measured for mixtures of modified and native deoxy-Hb S, DMA-modified monomeric deoxy-Hb S shortened the delay time prior to aggregation of native deoxy-Hb S. The other modified deoxy-Hb S did not affect the delay time, suggesting that these modified oligomeric hemoglobins neither participate in the formation of nuclei nor copolymerize with native deoxy-Hb S.     

Solution Radioactivated by Hadron Radiation Can Increase Sister Chromatid Exchanges

When energetic particles irradiate matter, it becomes activated by nuclear reactions. Radioactivation induced cellular effects are not clearly understood, but it could be a part of bystander effects. This investigation is aimed at understanding the biological effects from radioactivation in solution induced by hadron radiation. Water or phosphate buffered saline was activated by being exposed to hadron radiation including protons, carbon- and iron-ions. 1 mL of radioactivated solution was transferred to flasks with Chinese hamster ovary (CHO) cells cultured in 5 mL of complete media. The induction of sister chromatid exchanges (SCE) was used to observe any increase in DNA damage responses. The energy spectrum and the half-lives of the radioactivation were analyzed by NaI scintillation detector in order to identify generated radionuclides. In the radioactivated solution, 511 keV gamma-rays were observed, and their half-lives were approximately 2 min, 10 min, and 20 min. They respectively correspond to the beta+ decay of ¹⁵O, ¹³N, and ¹¹C. The SCE frequencies in CHO cells increased depending on the amount of radioactivation in the solution. These were suppressed with a 2-hour delayed solution transfer or pretreatment with dimethyl sulfoxide (DMSO). Our results suggest that the SCE induction by radioactivated solution was mediated by free radicals produced by the annihilated gamma-rays. Since the SCE induction and DMSO modulation are also reported in radiation-induced bystander effects, our results imply that radioactivation of the solution may have some contribution to the bystander effects from hadron radiation. Further investigations are required to assess if radioactivation effects would attribute an additional level of cancer risk of the hadron radiation therapy itself.     

Global and temporal regulation of gene expression in Xenopus kidney cells in response to presumed microgravity generated by 3D clinostats.

We documented changes in morphology and gene expression of the renal epithelial cell line A6 derived from Xenopus leavis adult kidney induced by long-term culturing with three dimensional clinostats. An oligo microarray analysis on A6 cells showed that mRNA levels of 52 out of 8091 genes were significantly altered in response to clinorotation. Upregulation or downregulation of gene expression became evident on day 8 and day 10 while there was no significant change on day 5. However, on day 15, expression of 18 out of 52 genes resumed to the levels similar to its original levels while remaining 33 genes maintained altered levels of expression. Quantitative analyses of gene expression by real-time PCR confirmed that changes in mRNA levels of selected genes were found only under clinorotation but not under hypergravity (7 G) and ground control (1 G). Morphological changes including loss of dome-like structures, disassembly of E-cadherin adherence junctions and disassembly of cortical actin were also observed over 10 days of culturing with clinorotation. The results revealed genes which expression was altered specifically in A6 cells cultured under clinorotation.     

Utilization of phosphate diester by the marine diatom Chaetoceros ceratosporus

Utilization of phosphate diester (PDE) and phosphodiesterase(PDEase) production by five marine phytoplankton species wereexamined in the laboratory to evaluate the contribution of PDEto the growth of phytoplankton. Among the five marine phytoplanktonspecies tested, only Chaetoceros ceratosporus was able to usethe PDE compound, bis(p-nitrophenyl) phosphate (bis-NPP), effectivelyas a sole phosphorus source. In addition, C. ceratosporus simultaneouslyproduced both PDEase and alkaline phosphatase (APase) at almostequal activity levels under the phosphate-deficient condition.These results suggest that PDE compounds presumably play animportant role as a phosphorus source for PDEase-producing phytoplanktonin coastal environments.     

Effect of amino acid at the beta 6 position on surface hydrophobicity, stability, solubility, and the kinetics of polymerization of hemoglobin. Comparisons among Hb A (Glu beta 6), Hb C (Lys beta 6), Hb Machida (Gln beta 6), and Hb S (Val beta 6)

Surface hydrophobicity, stability, solubility, and kinetics of polymerization were studied using hemoglobins with four different amino acids at the beta 6 position: Hb A (Glu beta 6), Hb C (Lys beta 6), Hb Machida (Gln beta 6), and Hb S (Val beta 6). The surface hydrophobicity increased in the order of Hb C, Hb A, Hb Machida, and Hb S, coinciding with the hydrophobicity of the amino acid at the beta 6 position. Solubility of the oxy-form of these hemoglobins decreased in relation to increases in their surface hydrophobicity, suggesting that the solubility is controlled by the strength of hydrophobicity of the amino acid at the beta 6 position. The solubility of the oxy-form of these hemoglobins is always higher than that of the deoxy-form. There is a similar linear relationship between the solubility and surface hydrophobicity among deoxyhemoglobins A, C, and Machida. However, the solubility of deoxy-Hb S deviated significantly from the expected value, indicating that the extremely low solubility of deoxy-Hb S is not directly related to the hydrophobicity of the beta 6 valine. Kinetic studies on the polymerization of deoxy-Hb Machida revealed a distinct delay time prior to polymerization. This confirms our previous hypothesis that beta 6 valine is not responsible for the delay time prior to gelation. The kinetics of the polymerization of 1:1 mixtures of sickle and non-sickle hemoglobins were similar to those of pure Hb S, suggesting that only one of the two beta 6 valines is involved in an intermolecular contact. In mixtures of equal amounts of Hb S and Hb A, Hb C, or Hb Machida, half of the asymmetrical AS, SC, and S-Machida hybrid hemoglobins behaved like Hb S during nucleation, while the other half behaved like the non-sickle hemoglobin.     

Inherited hydrocephalus in Csk: Wistar-Imamichi rats; Hyd strain: a new disease model for hydrocephalus

Hydrocephalic rats were found in a breeding colony of Csk: Wistar-Imamichi strain rats. In males, the hydrocephalus were serious and could be detected from 7 days after birth. Survival was 3-4 weeks. In females, the hydrocephalus was moderate, there was no abnormal external appearance, and the rats were able to mature. Ventricular dilatation was excessive in males but moderate in females. The total frequency of hydrocephalus was 34.3% in both males and females. Breeding data indicated that this disease is heritable and is single dominant and X-linked (symbol, Hyd). The female moderate hydrocephalics could be detected by progeny tests without examining brain sections. No evidence of developmental anomaly was observed in the ventricles. This hydrocephalus was classified as being of the communicating type, and this strain was named the Hyd strain as an animal model for human hydrocephalus.     

A method for detecting the optimum day for mating during the 4-day estrous cycle in the rat; measuring the value of electrical impedance of the vagina

A method for detecting the optimum day for mating in the rat was investigated. Cyclic changes of electrical impedance of vagina (EIV) were studied in the rats. EIV indicated high value (over 3,000 omega) only at proestrus, and it was lower (under 3,000 omega) at other stage of estrous cycle. These results apparently indicate that measuring the EIV made to distinguish proestrus from other phases of estrous cycle. The female caged with male showed high copulation rate (88 approximately 96%) when her EIV had been over 3,000 omega.