STUDIES ON COMPONENTS OF RESPIRATORY SYSTEM OF BEGONIA LEAVES

Investigations were made of the properties of diaphorase, cytochromec reductases, cytochrome c oxidase, and other components ofelectron transfer system in various fractions of leaf homogenateof Begonia semperflorens.

1. All the fractions tested showed the existence of cytochromec oxidase, succinic- and reduced diphosphopyridine nucleotide-cytochromec reductases, and diaphorase. Activities of these enzymes werefound to be associated mainly with the particulate fractions.The particulate fractions showed, in particular, a capacityof reducing oxidized cytochrome c with fumarate, malate, -ketoglutarate,ß-hydroxy-butyrate, and citrate.
2. Optimum pH foroxidation of cytochrome c by the particulatefractions was foundto be 5.5, while that for reduction was7.2.
3. The activityof cytochrome c reductase was partially suppressedby malonate.Partial inhibition of cytochrome c oxidase wascaused by azideand cyanide, the inhibitory effects observedbeing strongerwith particulate fractions than with solublefractions.

(Received August 11, 1962; )     

Involvement of Nitric Oxide in UVB‐Induced Pigmentation in Guinea Pig Skin

Ultraviolet (UV) B irradiation evokes erythema and delayed pigmentation in skin, where a variety of toxic and modulating events are known to be involved. Nitric oxide (NO) is generated from l ‐arginine by NO synthases (NOS). Production of NO is enhanced in response to UVB‐stimulation and has an important role in the development of erythema. NO has recently been demonstrated as a melanogen which stimulates melanocytes in vitro, however, no known in vivo data has been reported to support this finding. In this study, we investigated the contribution of NO with UV‐induced pigmentation in an animal model using an NOS inhibitor. UVB‐induced erythema in guinea pig skin was reduced when an NOS inhibitor, l ‐NAME (N‐nitro‐ l ‐arginine methylester hydrochloride), was topically applied to the skin daily, beginning 3 days before UVB‐irradiation. Delayed pigmentation and an increased number of DOPA‐positive melanocytes in the skin were markedly suppressed by sequential daily treatment with l ‐NAME. Furthermore, melanin content 13 days after UVB‐irradiation was significantly lower in skin treated with l ‐NAME than in the controls. In contrast, d ‐NAME (N‐nitro‐ d ‐arginine methylester hydrochloride), an ineffective isomer of l ‐NAME, demonstrated no effect on these UV‐induced skin responses. These results suggest that NO production may contribute to the regulation of UVB‐induced pigmentation.     

A Three-Dimensional Musculoskeletal Model of the Human Knee Joint. Part 1: Theoretical Construction

A three-dimensional model of the knee is developed to study the interactions between the muscles, ligaments, and bones during activity. The geometry of the distal femur, proximal tibia, and patella is based on cadaver data reported for an average-size knee. The shapes of the femoral condyles are represented by high-order polynomials: the tibial plateaux and patellar facets are approximated as flat surfaces. The contacting surfaces of the femur and tibia are modeled as deformable, while those of the femur and patella are assumed to be rigid. Interpenetration of the femur and tibia is taken into account by modeling cartilage as a thin, linear, elastic layer, mounted on rigid bone. Twelve elastic elements describe the geometry and mechanical properties of the cruciate ligaments, the collateral ligaments, and the posterior capsule. The model is actuated by thirteen musculotendinous units, each unit modeled as a three-element muscle in series with tendon. The path of each muscle is approximated as a straight line, except where it contacts and wraps around bone and other muscles; changes in muscle paths are taken into account using data obtained from MRI. In the first part of this paper, the model is used to simulate passive knee flexion. Quantitative comparisons of the model results with experimental data reported in the literature indicate that the relative movements of the bones and the geometry of the ligaments and muscles in the model are similar to those evident in the real knee. In Part II, the model is used to describe knee-ligament function during anterior-posterior draw, axial rotation, and isometric knee-extension and knee-flexion exercises.     

DIFFERENTIATION OF PARTIALLY MESODERMALIZED ECTODERM; HOMOIOGENETIC AND HETEROGENETIC INDUCTION BY PRIMARILY INDUCED PART OF THE ECTODERM

Using embryos of the Japanese newt, Cynops pyrrhogaster , homoiogenetic and heterogenetic induction were investigated in the partially mesodermaelzed presumptive ectoderm. Half of the isolated presumptive ectoderm was placed in contact with the swimbladder of the crucian carp, Carasius auratus , for 15 or 60 min, while the other half was stained with Nile blue sulfate at the same time. The distribution of the stained cells in the tissues evoked in the explants was examined after cultivation for 10 days.
Some mesodermal tissues were composed of both stained and unstained cells. This indicates homoiogenetic induction by the primarily induced part of the ectoderm on the other half. The neural and epidermal tissues in the explants were composed of stained cells only, except in one case. We conclude that the neural tissues are derived from cells not placed in contact with the swimbladder and that they are induced by the primarily induced part of the ectoderm.     

Changes in Sugar Content during Cold Acclimation and Deacclimation of Cabbage Seedlings

The relationship between freezing tolerance and sugar contentin cabbage seedlings was investigated. Seedlings exposed tonon-freezing low temperature (5 °C) acquired freezing tolerancedown to -6 °C. The degree of freezing tolerance increasedwith duration of exposure to low temperature (up to 10 d). Sucrose,glucose, fructose and myo -inositol were detected as solublesugars in cabbage leaves, and all soluble sugars, except formyo -inositol, and starch increased gradually during cold acclimationsuch that their levels were positively correlated with the degreeof freezing tolerance. The induced freezing tolerance was attributednot to ontogenetic changes but to cold acclimation. However,the induced freezing tolerance was lost after only 1 d of deacclimationat control temperatures, and this change was associated witha large reduction in sugar content. These results reveal that the sugar content of cabbage leavesis positively correlated with freezing tolerance. Brassica oleracea L.; cabbage; cold acclimation; deacclimation; freezing tolerance; sugars     

Purification of Acetylcholinesterase from Sea Urchin (Hemicentrotus pulcherrimus) Embryos by Affinity Chromatography

Only one form of acetylcholinesterase (AchE) was detected in Hemicentrotus pulcherrimus embryos. In H. pulcherrimus embryos as well as in the other sea urchin embryos, AchE activity begins to increase rapidly after gastrula stage.
Purification of AchE from plutei has been carried out by the procedure including affinity chromatography. Purified AchE had the activity 14,600 times higher than that of homogenate, and the final yield of AchE was 8%. The enzyme seems to be electrophoretically homogeneous, and has a molecular weight of 3 × 10⁵ as determined by Sepharose CL–6B column chromatography.     

Microtubule Formation in Regenerating Terminal Buds of the Silurid Fish, Corydoras aeneus

Regenerating terminal buds of Corydoras aeneus were observed by electron microscopy to determine how terminal buds developed with respect to microtubule formation. After surgical removal of the fish barbel, it and the terminal bud began to regenerate 1.5 weeks later at 25°C. The regenerating terminal buds were ovoid in shape and contained three types of cells. The first type of cell had extended cellular processes which contained numerous microtubules and tubules. A bundle of three or four microtubules ran parallel to the long axis of the cellular process. Receptor villi protruded from the cell two weeks later, suggesting that it is a receptor cell. The second cell type, which appeared 1.5 weeks after barbel removal, had numerous microtubules oriented along the long axis of the cellular process; and numerous dense granules appeared two weeks later, suggesting that it is a supporting cell. The third type of cell observed was a basal cell without cellular processes. These results suggest that microtubule formation plays an important role in the elongation of regenerating terminal buds.     

Seasonal changes in wing dimorphism of the lygaeid bug Dimorphopterus japonicus (Heteroptera: Lygaeidae) in relation to environmental factors

The lygaeid bug Dimorphopterus japonicus Hidaka, which mainly feeds on a eulalia, Miscanthus sinensis, shows marked wing dimorphism of brachyptery and macroptery. Its production of macropters is stimulated in rearing conditions such as high temperature, long photoperiod and crowding during the nymphal stage. In this study, we investigated the seasonal prevalence in occurrence and the seasonal change in incidence of macroptery in D. japonicus for field populations in Okayama, western Japan. The results demonstrated that nymphal density was a key factor in determining the proportion of macropters, indicating an escape strategy from crowded populations. The field surveys also revealed that this bug has a univoltine life cycle in Okayama. There was a seasonal change in the incidence of macroptery in the new generation. The combined effects of temperature and photoperiod on wing‐form determination explained this seasonality trait.     

Potential effects of tyramine on the transition to reproductive workers in honeybees (Apis mellifera L.)

Abstract.  To explore the role of brain tyramine in reproductive worker honeybees, its effects after injection and oral treatment on brain dopamine levels and ovarian development in queenless worker honeybees are determined. Both tyramine injection and oral treatment in 10-day-old queenless bees leads to tyramine transportation into the brain and significantly elevates brain dopamine levels as a function of the tyramine concentration. Ovarian diameters are significantly larger in 10-day-old queenless bees treated with tyramine compared with queenless bees of the same age without tyramine treatment. Results on yolk formation in the ovary support the finding of increased ovarian diameter, suggesting that oral tyramine treatment accelerates ovarian development through dopamine effects and/or direct effects of tyramine on the ovary in queenless bees. Thus, tyramine has potential effects on the enhancement of brain dopamine levels and the acceleration of ovarian development for the transition of normal workers to reproductive worker honeybees.