Molecular cloning of equine chromogranin A and its expression in endocrine and exocrine tissues

Chromogranin A (CGA) is a member of a family of highly acidic proteins co-stored and co-released with catecholamines in the adrenal medullary cells as well as in other neurons and paraneurons. The nucleotide sequence encoding equine CGA was determined using RT-PCR and rapid amplification of complementary DNA (cDNA) ends (RACE) techniques. A total 1,828 bp of the nucleotide sequence reveals that equine CGA is a 448-residue protein preceded by an 18-residue signal peptide. Comparison of the amino acid sequence of equine CGA with those of human, porcine, bovine, mouse, rat and frog CGA showed high conservation at the NH2-terminal 1-77 amino acids regions (94.8%, 93.5%, 92.2%, 81.8%, 83.1% and 66.2%, respectively) and COOH-terminal 314-430 amino acids regions (90.6%, 81.4%, 90.6%, 80.5%, 83.3% and 39.0%, respectively), as well as a potential dibasic cleavage site, whereas the middle portion showed marked sequence variation (52.5%, 49.1%, 38.9%, 26.6%, 27.9% and 6.2%, respectively). Northern blot analysis and RT-PCR elucidated the tissue distribution of equine CGA mRNA. Its expression was confirmed not only in the adrenal medullary cells but also in other organs (cerebrum, cerebellum, pituitary gland, spinal cord, liver, thyroid gland, striated muscle, lung, spleen, kidney, parotid gland and sublingual gland). Further, in adrenal chromaffin cells and pituitary cells of the anterior-intermediate lobe, the expression was confirmed by in situ hybridization with anti-sense CGA cRNA probe.     

Effects of in vivo administration of anti-IL-10 or anti-IFN-gamma monoclonal antibody on the host defense mechanism against Plasmodium yoelii yoelii infection

Our previous reports indicated that C57BL/6 mice infected with a lethal variant of Plasmodium yoelii 17X (P. yoelii 17XL) produced high levels of interleukin 10 (IL-10) and interferon-gamma (IFN-gamma) while mice infected with the nonlethal variant of the parasite did not produce detectable levels of IL-10. In the present study, the involvement of IL-10 and IFN-gamma in exacerbation or regulation of blood-stage malaria was investigated by using the lethal variant of P. yoelii 17XL and monoclonal antibodies (mAb) against the cytokines. C57BL/6 mice were injected intraperitoneally with a neutralizing anti-IL-10 mAb or anti-IFN-gamma mAb after inoculation with P. yoelii 17XL. Treatment of mice with anti-IL-10 mAb resulted in substantial prolongation of survival and 60% of treated mice survived while 100% of control mice died by day 11. On the contrary, treatment of mice with anti-IFN-gamma mAb exacerbated infection and all mice died after an earlier period than those treated with normal rat Ig. No differences in parasitemias were found between treated and untreated mice. To elucidate the involvement of nitric oxide in the host protection or exacerbation, mice were treated with aminoguanidine, an inhibitor of nitric oxide synthetase, after inoculation of P. yoelii 17XL. Neither mortality nor parasitemia was influenced by the treatment. These results indicate that an IFN-gamma response is associated with protective immunity in mice infected with P. yoelii 17XL, while an IL-10 response is associated with disease exacerbation during the infection.     

Larger body size with higher predation rate

Abstract –  Many studies have reported that prey fish increase in body size with an increase in piscivore abundance, and this change may be explained mainly by two processes: release from intraspecific competition because of a reduction in the prey fish population and size-dependent selection by the piscivore. However, the evolutionary mechanism is not yet fully understood. Here, we first show the body-size dynamics of the semelparous gobiid fish Gymnogobius isaza , which is one of the prey fishes of the introduced largemouth bass, Micropterus salmoides . The data show the possibility that the goby body size might have changed in parallel to the number of largemouth bass from 1975 to 2002, which may be one of the best examples of the positive relationship between body size and predation rate. Furthermore, we explored a mathematical model to consider a body-size change of semelparous fish from an evolutionary viewpoint. The model provided the following qualitative predictions: (1) the optimal assimilation rate increases with predation pressure; (2) a rapid large-scale increase in predation pressure may exterminate the prey fish because of delayed evolution of the assimilation rate; and (3) the optimal assimilation rate increases when the prey fish is more likely to die through predation than through natural mortality or when it is difficult to grow larger because of low resource availability or insufficient foraging efficiency. Moreover, we propose a theoretical framework to evaluate the relative effects of ecological and evolutionary processes over the long term.     

Interactive effects of leaf age and self-shading on leaf structure,photosynthetic capacity and chlorophyll fluorescence in the rain forest tree,Dryobalanops aromatica

In the tropical canopy tree, Dryobalanops aromatica Gaertn. f., upper-canopy leaves (UL) develop under sunlit conditions but are subjected to self-shading within the crown as they age. In contrast, lower-canopy leaves (LL) are exposed to uniform dim light conditions throughout their life span. By comparing leaf morphology and physiology of UL and LL, variations in leaf characteristics were related to leaf age and self-shading. Mass-based chlorophyll (chl) concentration and the chlorophyll/nitrogen (chl/N) ratio were lower and the chl a/b ratio was higher in UL than in LL. In UL, the chl/N ratio gradually increased and the chl a/b ratio gradually decreased with leaf aging, whereas these ratios remained unchanged with leaf age in LL. The effective quantum yield of photosystem II (PSII) (DeltaF/F(m)') at a given irradiance remained unchanged with leaf age in LL, whereas DeltaF/F(m)' changed with leaf age in UL. These data indicate N reallocation within the leaves from carbon fixation components to light harvesting components and a dynamic regulation of photochemical processes of PSII in response to increased self-shading of UL. Despite the difference in light environment with leaf age between UL and LL, maximum photosynthetic rates and nitrogen-use efficiency decreased with leaf aging in both UL and LL. Constancy in the chl/N ratio with leaf age in LL indicated that the decrease in photosynthetic capacity was caused by effects other than shading, such as leaf aging. We conclude that N reallocation and acclimation of PSII to self-shading occurred even in mature leaves, whereas the change in photosynthetic capacity with leaf age was more conservative.     

Dynamic excitation and static loading tests of glulam lattice floor

A wooden lattice floor with high stiffness and damping capacity has been developed to solve noise problems in wooden apartment houses. The lattice floor consisted of Douglas fir glulam beams with inserted steel plate joints and drift pins. To examine the structural performance of the floor, dynamic excitation and static loading tests were conducted on the full size floor. The first and second order resonance frequencies of the floor were 13.5Hz and 27.0Hz, respectively. These frequencies are similar to the peak frequency of a conventional wooden floor and the combined floor fabricated from glued laminated timber and iron. The maximum static load of the floor was 127kN. The apparent flexural rigidity was less than half the value of several floors studied in the past. However, it is considered that the stiffness is improved by constructing panels and this floor has almost equivalent performance. Relative deflection was not affected by the loading history.Part of this study was presented at the International Wood Engineering Conference, New Orleans, October 1996.     

The regeneration characteristics of Picea jezoensis and Abies sachalinensis on cut stumps in the sub-boreal forests of Hokkaido Tokyo University Forest

The regeneration characteristics of Picea jezoensis Carr. and Abies sachalinensis Masters on cut stumps in natural forest under selection cutting and natural regeneration were studied in a sub-boreal forest of the Hokkaido Tokyo University Forest. The following points were investigated: (1) what decay class of cut stumps are suitable for regeneration of the two conifer species; (2) what part, either a root collar or a cut surface, of a cut stump is suitable for regeneration; and (3) what percentages of cut stumps support conifer seedlings. The percentages of cut stumps that supported conifer seedlings increased as the decay class advanced, but slightly decreased in the latest stage of decay class, V. Both P. jezoensis and A. sachalinensis seedling densities per cut stump and per square meter of cut stumps increased as the decay class advanced. Some root collars in decay class IV supported A. sachalinensis seedlings, but few supported P. jezoensis. Densities of P. jezoensis per cut surface and per square meter of cut surface in decay class IV were higher than those of A. sachalinensis. On the other hand, densities of A. sachalinensis per root collar and per square meter of root collar in decay class IV were higher than those of P. jezoensis. Cut stumps in advanced decay classes are more suitable for regeneration of P. jezoensis and A. sachalinensis than those in early decay classes. Whereas a cut surface is more suitable for regeneration of P. jezoensis, a root collar is more suitable for regeneration of A. sachalinensis. Cut stumps are not sufficient to ensure enough conifer regeneration because not all conifer cut stumps in advanced decay classes support conifer seedlings.     

Structure and regeneration dynamics of dominant tree species along altitudinal gradient in a dry valley slopes of the Bhutan Himalaya

The aim of our study was to characterize the structure and regeneration dynamics of dominant tree species along altitudinal gradient in dry valley slopes of the Bhutan Himalaya. In the typical dry valley slopes of west-central Bhutan, we analyzed forest stratification, tree crown projection, and population structure from lower dry valley bottom (1520 m a.s.l.) to upper humid ridge top (3370 m a.s.l.) across five major forest types: i.e. (1) dry pine forest with Pinus roxburghii as a dominant species; (2) mixed broad-leaved forest with Quercus lanata (evergreen), Quercus griffithii, (deciduous), and Rhododendron arboreum (evergreen) as major canopy species; (3) evergreen broad-leaved forest dominated by Quercus semecarpifolia, Quercus glauca and Quercus oxyodon; (4) cool conifer forest dominated by Abies densa and Tsuga dumosa; (5) cold conifer forest with A. densa and Juniperus recurva as dominants. In general, regeneration pattern of major dominant species shifted from inverse-J (lower altitudes), to sporadic (mid-altitudes), and to uni-modal type (upper altitudes) corresponding to three regeneration trends: (1) invasive at the lower, warm, dry forest under relatively strong human disturbances; (2) stable/balanced at the mid-altitude, in a relatively stable, mature moist evergreen broad-leaved forest with gap regeneration; and (3) poor/low regeneration at the upper, cool, humid conifer forest with a continuously cattle-grazed understory. Overall, regeneration patterns were balanced in all forest types, however care should be taken to prevent excessive exploitation of dry pine forest of lower valley bottom, and to control cattle grazing at higher altitudes. Our findings can be incorporated into management plans for sustainable management and conservation of mountain forests.     

Effect of moisture content of members on mechanical properties of timber joints

To investigate the effect of moisture content (MC) of members on the mechanical properties of timber joints, bending tests of precut joints and shear tests of dowel-type joints were carried out using timbers of Japanese cedar (Cryptomeria japonica D. Don) with three moisture conditions: green, kiln-dried with a MC target of 15%, and over-kiln-dried with a MC target of 5%. For the bending test, timbers were processed with a precut processing machine into “koshikake-ari” (a kind of dovetail joint) and “koshikake-kama” (a kind of mortise and tenon joint). A pair of members was jointed together without mechanical fasteners. Bolts (diameter = 12 mm) and nails (diameter = 2.45 mm) were used as dowels in the shear test. Bolted joints were constructed with one bolt and two metal side plates. Two nails and two metal side plates were used for the nailed joint. For precut joints, no clear effect of MC was recognized on maximum moment and initial stiffness. The maximum strength of mechanical joints assembled with kiln-dried wood was changed by the degree of drying. Stiffness of the joints assembled with kiln-dried specimens was larger than that of the joints assembled with green specimens.Part of this study was presented at the 7th International IUFRO Wood Drying Conference, Tsukuba, July 2001     

Effects of current-year and previous-year PPFDs on shoot gross morphology and leaf properties in Fagus japonica

We investigated how shoot gross morphology and leaf properties are determined in Fagus japonica Maxim., a deciduous species with flush-type shoot phenology, in which all leaves are produced in a single flush at the start of each season. We examined relationships between current-year shoot properties and local light environment in a 14-m tall beech tree growing in a deciduous forest. Leaf number (LN), total leaf area (TLA), and total leaf length (SL) of the current-year shoot increased with increasing photosynthetic photon flux density (PPFD). Leaf thickness, dry mass per leaf area and nitrogen content on a leaf area basis increased, whereas the chlorophyll/N ratio decreased with increasing PPFD. To separate the effects of current-year PPFD from those of previous year(s), we artificially shaded a part of the uppermost leaf tier. Reciprocal transfers of beech seedlings between controlled PPFD regimes were also made. Characteristics of shoot gross morphology such as LN, TLA and SL were largely determined by the PPFD of the previous year. The exception was the length of the longest "long shoots" with many leaves, in which elongation appeared to be influenced by both previous-year and current-year PPFD. In contrast, leaf properties were determined by current-year PPFD. The ecological implications of our findings are discussed.