STIMULATION OF COLLAGEN SYNTHESIS IN RAT CARDIAC FIBROBLASTS BY EXPOSURE TO HYPOXIC CULTURE CONDITIONS AND SUPPRESSION OF THE EFFECT BY NATRIURETIC PEPTIDES

Synthesis of type I and type III collagens by rat cardiac fibroblasts was stimulated when the cells were cultured under 95% N₂/5% CO₂for one hour followed by incubation under normoxic conditions for 24 hours. The stimulative effect was attenutated by the presence of atrial natriuretic peptide (ANP, 10⁻⁶m) or brain natriuretic peptide (BNP, 10⁻⁶m) in the culture medium. Northern blot analysis indicated that α1(I) and α1(III) collagen mRNA levels were also increased by hypoxia, and decreased with the addition of ANP or BNP in a dose-dependent manner. These results indicate interaction between intracellular signals of a physical stimulus (hypoxic stress) and those of a chemical one (ANP or BNP) and demonstrate that both signals regulate collagen synthesis by cardiac fibroblasts at the levels of the mRNAs. The results also suggest that natriuretic peptides produced by cardiomyocytesin vivomay function as paracrine factors that play a role in the prevention of cardiac fibrosis in ischaemic heart diseases.     

Notes on the biology of the Oriental amblyoponine ant Myopopone castanea: Queen‐worker dimorphism,worker polymorphism and larval hemolymph feeding by workers (Hymenoptera: Formicidae)

Queen‐worker dimorphism, worker polymorphism and worker behavior in the Oriental amblyoponine ant Myopopone castanea were studied. Queen body size was large (head width, 3.0 mm) with 24 to 32 ovarioles while workers showed a remarkable size variation in both body size (head width 1.48 to 2.18 mm) and ovariole number (6 to 22). Both head width and abdomen width showed allometric growth against thorax width. Workers performed larval hemolymph feeding as was described for Amblyopone silvestrii queens.     

Effects of NO2 and O3 Alone or in Combination on Kidney Bean Plants (Phaseolus vulgaris L.): Products of 13CO2 Assimilation Detected by 13C Nuclear Magnetic Resonance

The effect of exposure of kidney bean primary leaves to NO₂and O₃, alone or in combination, on the fate of ¹³CO₂ assimilatedby photosynthesis was examined by ¹³C-NMR. There were more than70 peaks appearing in the ¹³C-NMR spectra for substances extractedfrom leaves with 80% ethanol. The 16 relatively well resolvedpeaks corresponded to signals from three sugars, two organicacids and four amino acids. These signals were used to estimatepool sizes and ^l3C incorporation. Exposures to NO₂ and O₃ increased the amounts of sucrose andfructose, but not the incorporation of the ¹³C label during10 min photosynthesis from ¹³CO₂. This suggests the presenceof photo-synthetically inactive pools of sucrose and fructose.Amounts of glycine and serine, and ¹³C incorporation into them,were increased by the exposure to the pollutants. The incorporationof ¹³C into alanine was stimulated by exposure to NO₂, but notby exposure to O₃ alone. The present study shows that with only simple procedures ofsample preparations ¹³C-NMR provides information on the productsof photosynthesis in leaves stressed by the two air pollutants. Key words: NO₂, O₃, Phaseolus vulgaris, CO₂ assimilation, ¹³C-NMR     

Comparative Analysis of Hair Melanins by Chemical and Electron Spin Resonance Methods

Eighteen hair samples from Karakul newborn lambs with various colors were estimated for eumelanin and pheomelanin contents (C_e and C_p, respectively) by electron spin resonance (ESR) spectrometry and by high-performance liquid chromatography (HPLC). Correlation coefficients between the values estimated by the ESR and HPLC methods were 0.96, 0.93, and 0.99 for C_e, C_p, and C_e/C_p, respectively. The high correlation coefficients show that both methods fit well for estimation of relative values of these parameters. The absolute values of C_e and C_e/C_p coincide rather well when C_e is high, but considerable discrepancies appear when C_e is low. The reasons for these discrepancies are discussed. The HPLC method appears to be more sensitive for detection of low concentrations of pheomelanin, while the ESR method fits well for mass selection purposes.     

Histone modification and signalling cascade of the dormancy‐associated MADS‐box gene,PpMADS13‐1, in Japanese pear (Pyrus pyrifolia) during endodormancy

Dormancy‐associated MADS‐box (DAM) genes play an important role in endodormancy phase transition. We investigated histone modification in the DAM homolog (PpMADS13‐1) from Japanese pear, via chromatin immunoprecipitation–quantitative PCR, to understand the mechanism behind the reduced expression of the PpMADS13‐1 gene towards endodormancy release. Our results indicated that the reduction in the active histone mark by trimethylation of the histone H3 tail at lysine 4 contributed to the reduction of PpMADS13‐1 expression towards endodormancy release. In contrast, the inactive histone mark by trimethylation of the histone H3 tail at lysine 27 in PpMADS13‐1 locus was quite low, and these levels were more similar to a negative control [normal mouse immunoglobulin G (IgG)] than to a positive control (AGAMOUS) in endodormancy phase transition. The loss of histone variant H2A.Z also coincided with the down‐regulation of PpMADS13‐1. Subsequently, we investigated the PpMADS13‐1 signalling cascade and found that PpCBF2, a pear C‐repeated binding factor, regulated PpMADS13‐1 expression via interaction of PpCBF2 with the 5′‐upstream region of PpMADS13‐1 by transient reporter assay. Furthermore, transient reporter assay confirmed no interaction between the PpMADS13‐1 protein and the pear FLOWERING LOCUS T genes. Taken together, our results enhance understanding of the molecular mechanisms underlying endodormancy phase transition in Japanese pear.     

Clustered Genes Require Extragenic Territorial DNA Sequences

This paper is concerned with the basic question as to whether there exists a complex interaction between DNA sequences which have little specific function and functional genes regarding the spatial arrangement of the gene. Since gene clusters are a characteristic and basic feature of gene structure in higher eukaryotes, the size of extragenic DNA sequences surrounding the individual genes of various clustered gene families were compared. The size of the intergenic region, which is composed of the extragenic DNA sequences flanking the 3'-end of one gene and those flanking the 5'-end of the other gene, of the paired genes increases as the genes becomes larger. However, such a gene size-dependent increase is not seen if the total gene size of the paired genes is less than 0.3 kb or greater than 4 kb. The results suggests that a higher eukaryote gene requires extragenic territorial DNA sequences surrounding it, which presumably are necessary to maintain the gene's active functions.