Studies on the Protection Test of Pertussis Vaccine. Immunization Period and Protectivity

In order to confirm the data reported in the previous papers, variously prepared pertussis vaccines were employed in the present investigation. Pertussis organisms grown either on a solid or in a liquid semisynthetic medium were treated by: (1) heating at 56 C for 30 min, (2) storage in 0.1% formalin at 37 C for 5 days, (3) storage in 0.1% formalin at 25 C for 5 days, and (4) simple addition of sodium ethyl-mercuri thiosalicylate (merthiolate) as a preservative. Freeze-dried vaccines were made from these preparations four and a half months later. Mice were immunized intraperitoneally and challenged intracerebrally with a virulent strain of Bordetella pertussis 10 or 17 days later. The data were analyzed statistically assuming the probit corresponding to the percentage of survivors at any dose be a linear function of the logarithms of the dose. The 50% effective doses (ED₅₀) and slopes of each vaccine were found to be uniform. More accurate estimates of ED₅₀ were obtained by employing a pooled slope in each experiment. From these ED₅₀ values, the relative potency was estimated by comparing the value of a vaccine to that of a dried merthiolate-vaccine. For vaccines derived from solid cultures, with an immunization period of 17 days, the relative potency of the vaccine heated at 56 C was 0.63 (95% fiducial limits=0.52 to 0.76); the value for the formalinized vaccine at 37 C was 0.40 (0.30 to 0.53) and one at 25 C was 0.51 (0.34 to 0.77). Vaccines derived from liquid cultures showed a relative potency of 20 to 50% less than that of corresponding vaccine derived from a solid culture. The potency obtained for the 17 day immunization period was usually higher than that for the 10 day period. Using the overall-pooled slope, an experimental design which will be appropriate for statistical analysis is discussed.     

Purification and characterization of a novel chitinase from Burkholderia cepacia strain KH2 isolated from the bed log of Lentinus edodes,Shiitake mushroom

One of the chitinases secreted in the culture filtrate of a gram-negative bacteria, Burkholderia cepacia strain KH2, which was isolated from the bed log of Lentinus edodes, Shiitake mushrooms, was purified by DEAE Sepharose CL-6B chromatography, followed by Sephacryl S-100 HR gel filtration. The purified enzyme was homogenous, determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), with an estimated molecular weight of 34,000 and an isoelectric point (pI) of 5.9. The enzyme was stable at pH values of 4.0-6.0, and at temperatures up to 50 degrees C; the optimum pH and temperature were 4.5 and 50 degrees C, respectively. The enzyme exhibited higher activities toward chitosan 7B, a 62% deacetylated chitosan, than toward the highly deacetylated chitosan substrates. The enzyme was observed to drastically hydrolyze partially deacetylated chitin substrates, with the subsequent formation of N-acetylchitooligosaccharides [(GlcNAc) (n), n=2-7]. Separation and quantification of the hydrolysis products of (GlcNAc) (n), n52-6, by HPLC showed the splitting into (GlcNAc)(n), n=3-6. Activity toward N-acetylchitobiose was not detected. Oligomers with a higher number of units than the starting substrate were also detected, which indicate transglycosylation activity.     

Stem turnover strategy of multiple-stemmed woody plants

We have investigated stem turnover strategy for Lindera umbellata, an understory shrub that sprouts from its rootstock under natural conditions to replace constituent stems, on the basis of the hypothesis that the multiple-stemmed form of woody species is an adaptation enabling efficient reproduction in high-stress environments. We tested the hypothesis that the timing of stem replacement maximizes sexual reproduction for the shrub. We developed a model for the time of optimum replacement of a stem by a daughter stem which maximizes the sexual reproduction of a shrub and tested the model using L. umbellata growing in the field. From the model, the optimum time of replacement of a stem with a daughter stem is when cumulative sexual reproduction per unit time for the stem is maximum. In practice, this will be the last age (t _opt) at which annual sexual reproduction in a stem can potentially exceed cumulative sexual reproduction per unit time for the stem. Half of the stems died at almost t _opt and had sexually mature daughter stems at that time. Other stems, however, died at times more remote from t _opt when daughter stems were sexually immature. It is thought that normal replacement of the latter stems was prevented by accidents such as breakage. We conclude that clumps of L. umbellata achieve efficient sexual reproduction by stem replacement at the optimum time, although accidents can, to some extent, determine when the stem actually dies.     

Comparison of the physiology, morphology, and leaf demography of tropical saplings with different crown shapes

Branch architecture, leaf photosynthetic traits, and leaf demography were investigated in saplings of two woody species, Homolanthus caloneurus and Macaranga rostulata, co-occurring in the understory of a tropical mountain forest. M. rostulata saplings have cylindrical crowns, whereas H. caloneurus saplings have flat crowns. Saplings of the two species were found not to differ in area-based photosynthetic traits and in average light conditions in the understory of the studied site, but they do differ in internode length, leaf emergence rate, leaf lifespan, and total leaf area. Displayed leaf area of H. caloneurus saplings, which have the more rapid leaf emergence, was smaller than that of M. rostulata saplings, which have a longer leaf lifespan and larger total leaf area, although M. rostulata saplings showed a higher degree of leaf overlap. Short leaf lifespan and consequent small total leaf area would be linked to leaf overlap avoidance in the densely packed flat H. caloneurus crown. In contrast, M. rostulata saplings maintained a large total leaf area by producing leaves with a long leaf lifespan. In these understory saplings with a different crown architecture, we observed two contrasting adaptation strategies to shade which are achieved by adjusting a suite of morphological and leaf demographic characters. Each understory species has a suite of morphological traits and leaf demography specific to its architecture, thus attaining leaf overlap avoidance or large total leaf area.     

Distribution of -amino-acid oxidase and -serine in vertebrate brains

In mammalian brains, -amino-acid oxidase activity is absent or scarce in the forebrain, is confined to the brain stem and cerebellum, and its localization is extended to the spinal cord. The oxidase-containing cells are astrocytes including Bergmann glial cells. Neither neurons, endothelial cells, oligodendrocytes nor ependymal cells show the oxidase activity. Free -serine, a potent activator of the N-methyl- -aspartate (NMDA) receptor, is in high levels in the forebrain (ca. 0.4 μmol/g wet weight), and in low levels in the hindbrain. Thus, the localization of the oxidase activity is inversely correlated with the distribution of -serine in mammalian brains. This inverse correlation is generally found in vertebrate brains. These results indicate that -amino-acid oxidase decomposes -amino acids including -serine in vertebrate brains, and that the magnitude of its activity is important in determining the regional concentrations of -amino acids in the steady states.     

Canopy ergodicity: can a single leaf represent an entire plant canopy?

While leaves typically emerge near shoot apices around the outer surface of a plant canopy, their relative position “moves” deeper into the canopy as additional leaves emerge. The photosynthetic capacity (A _max) of a given leaf can be expected to decline over time as its relative position (P _r) in the canopy becomes progressively deeper; this can be observed as a spatial gradient with the A _max of leaves declining distally from the shoot apex. As a consequence, we propose that the photosynthetic capacity averaged over a single leaf’s lifespan is equivalent to the average photosynthetic capacity of the entire plant canopy at a given time; in other words, there is an ergodic time to space averaging in the organization and development of plant canopies. We tested this “canopy ergodic hypothesis” in two woody (Alnus sieboldiana and Mallotus japonica) and two herbaceous (Polygonum sachalinensis and Helianthus tuberosus) species by following the photosynthetic capacity in 100 individual leaves from the time of their emergence until their death. We compared the average photosynthetic capacity of individual leaves through time (time-average) to the average photosynthetic capacity of all the leaves along a shoot at the time of emergence of the focal leaf (space-average). We found that A _max and P _r were positively correlated and that the time-averages of three plant species (Alnus, Mallotus, and Helianthus) were not significantly different from the corresponding space-averages, although the averages differed among individual plants. Polygonum, however, did show significant differences between time and space averages. Ergodicity appears to apply to the leaf–canopy relationship, at least approximately—the average photosynthetic capacity of a single leaf through time (time-average) can represent the average photosynthetic capacity of the entire canopy.     

Reduction of photosynthesis before midday depression occurred: leaf photosynthesis of <Emphasis Type="Italic">Fagus crenata</Emphasis> in a temperate forest in relation to canopy position and a number of days after rainfall

We investigated an effect of canopy position and a number of days after rainfall on reduction of photosynthetic rate in a Fagus crenata forest in summer 2008, during days when midday depression was not apparent. We compared in situ photosynthetic rate and photosynthetic rate that was calculated by photosynthetic light response curves measured in the morning. The ratio, in situ photosynthesis divided by the curve-estimated value, declined towards the end of each day for the upper leaves, but not for the lower leaves. Total photosynthesis was reduced only for the upper leaves by 12% during 5 days after the rainfall.     

Leaf Longevity as a Normalization Constant in Allometric Predictions of Plant Production

In metabolic scaling theory the size-dependence of plant processes is described by a power function of form Y=Y _o M ^θ where Y is a characteristic such as plant productivity that changes with plant size (M) raised to the θ ^th power and Y _o is a normalization constant that adjusts the general relationship across environments and species. In essence, the theory considers that the value of θ arises in the size-dependent relationship between leaf area and vascular architecture that influences plant function and that Y _o modulates this general relationship to account for ecological and evolutionary effects on the exchange of resources between plant and environment. Enquist and colleagues have shown from first principles that Y _o is a function of carbon use efficiency, the carbon fraction of a plant, the area-specific carbon assimilation rate of a leaf, the laminar area of a leaf, and the mass of a leaf. We show that leaf longevity provides a functional integration of these traits that can serve as a simpler normalization in scaling plant productivity for individual species and potentially for mixed-species communities as well.