Penetration of Eimeria tenella sporozoites under different oxygen concentrations in vitro.

Sporozoites of Eimeria tenella (Wisconsin strain) were inoculated onto monolayers of normal chicken kidney fibroblasts and cultured in RPMI-1640 supplemented with 5% fetal bovine serum, sodium bicarbonate, and gentamicin under either aerobic, 5% CO2/95% air, or anaerobic conditions. Penetration of fibroblasts by sporozoites under CO2 or anaerobic conditions at 2 and 24 hr postinoculation was 3-4 times greater than that in the aerobic atmosphere. Effect of reduced oxygen concentrations, i.e., 20.0, 12.5, and 5.0% oxygen, was also investigated in an N2-O2-CO2 incubator. Under 5.0 and 12.5% oxygen at 2 and 24 hr postinoculation, the number of sporozoites that penetrated was about 4 and 2 times greater, respectively, than under 20.0% O2. These results indicate that lower oxygen concentrations provide for greater penetration by E. tenella sporozoites in cultured cells.     

Ultrastructure of mass of floccular substance in the parathyroid gland of golden hamster.

Mass of floccular substance was observed in the parathyroid glands of fetal, newborn and infantile golden hamsters. Mass composed of floccular substances was spherical with no limiting membrane around it. It was located near the nucleus and the Golgi area, but was also observed in the peripheral cytoplasm. No cell organelles were detected within area of mass.     

Size dependent predatory pressure in the Japanese horned beetle,Allomyrina dichotoma L. (Coleoptera; Scarabaeidae)

We surveyed male survival and reproductive performances associated with dimorphism in the Japanese horned beetleAllomyrina dichotoma L. in a secondary forest in 1996. Morphological comparisons between living marked and prey individuals indicated that the larger horned males suffered higher predatory pressure than the smaller ones. The dominant predators of the beetles were suspected to be 2 crow species. The small-horned males showed lower recapture rates than the large-horned ones. This suggested that the former was more sensitive to disturbance, and/or dispersed more than the latter. Fighting behavior was rarely seen because of the low population density of the beetles in the study area. These results suggested that the large-horned males suffer not only the injury risk of intrasexual competition but also more predatory risk than the small-horned ones.     

Mechanism of poly(ethylene glycol) interaction with proteins

Poly(ethylene glycol) (PEG) is one of the most useful protein salting-out agents. In this study, it has been shown that the salting-out effectiveness of PEG can be explained by the large unfavorable free energy of its interaction with proteins. Preferential interaction measurements of beta-lactoglobulin with poly(ethylene glycols) with molecular weights between 200 and 1000 showed preferential hydration of the protein for those with Mr greater than or equal to 400, the degree of hydration increasing with the increase in poly(ethylene glycol) molecular weight. The preferential interaction parameter had a strong cosolvent concentration dependence, with poly(ethylene glycol) 1000 having the sharpest decrease with an increase in concentration. The preferential hydration extrapolated to zero cosolvent concentration increased almost linearly with increasing size of the additive, suggesting steric exclusion as the major factor responsible for the preferential hydration. The poly(ethylene glycol) concentration dependence of the preferential interactions could be explained in terms of the nonideality of poly(ethylene glycol) solutions. All the poly(ethylene glycols) studied, when used at levels of 10-30%, decreased the thermal stability of beta-lactoglobulin, suggesting that caution must be exercised in the use of this additive at extreme conditions such as high temperature.     

Preparation and characterization of recombinant DNA-derived human interferon-gamma

An attempt was made to prepare a highly purified, active recombinant DNA-derived human interferon-gamma. When the protein was denatured in urea and refolded, gel filtration and sedimentation velocity experiments indicated the presence of two forms, which are different in size and are not in a rapid reversible equilibrium. The two forms could be chromatographically separated. Far-UV circular dichroic spectra indicated the presence of secondary structures for both forms. Near-UV circular dichroic spectra revealed that the smaller form is folded into a rigid tertiary structure. The antiviral activity of the two forms of interferon-gamma showed a significant difference, i.e. the smaller form was 4-8-fold more active than the larger form. A variety of experiments show that the smaller form is more active, homogeneous, soluble, and stable than the larger form.     

Impaired generation of prostaglandins from isolated gastric surface epithelial cells in portal hypertensive rats

We studied generation of prostaglandins E2 and 6-keto F1a by surface epithelial cell isolated from the gastric mucosa of portal hypertensive and sham-operated rats. Oxygenated cell suspensions containing 80 +/- 3% of surface epithelial cells were incubated for 30 min at 37 degrees C and the concentration of prostaglandin E2 and 6-keto-prostaglandin F1a in medium was measured by radioimmunoassay. Viability of the cells was assessed with Fast green exclusion at baseline and after 30-min and 60-min incubation. Within 30 minutes the surface epithelial cells obtained from portal hypertensive rats generated 22.0 +/- 1.6 (mean +/- SE) pg prostaglandin E2 and 40.7 +/- 4.7 pg 6-keto prostaglandin F1a, per 10(6) cells. These were significantly less than prostaglandin generation by cells obtained from sham-operated rats. The viability of the surface epithelial cells from portal hypertensive rats was also significantly reduced compared with sham-operated rats after 60 minute incubation. Reduced ability of the surface epithelial cells to generate prostaglandins may be one mechanism for increased susceptibility of portal hypertensive gastric mucosa to injury by noxious agents.     

Photoregulation of Anthocyanin Synthesis in Apple Fruit under UV-B and Red Light

The involvement of photomorphogenic photoreceptors in anthocyaninsynthesis was investigated in apple fruits under UV light from280 to 320 nm (UV-B) and red light (R). Short-term R treatmentwas ineffective in the induction of anthocyanin synthesis butthe involvement of phytochrome was indicated by the resultsof long-term irradiation (18 h) with R. The inductive effectof 18 h UV-B on anthocyanin synthesis was stimulated synergisticallyby subsequent irradiation with R for 15 min, and the R, far-redlight (FR) photorevesibility of this effect indicated the involvementof phytochrome in this synergism. The effect of UV-B on anthocyaninsynthesis was not influenced by subsequent irradiation withFR, suggesting that the effect of UV-B was independent of phytochrome,and that a specific photoreceptor for UV-B was involved. WhenR was given simultaneously with UV-B (18 h), anthocyanin wassynthesized at a much higher rate than it was after sequentialirradiation with UV-B and R. Photosynthesis was shown to beinvolved inthis synergistic increase in the synthesis of anthocyanin,although the involvement of phytochrome in the expression ofthis response, at least in part, was suggested by a reductionin the rate of anthocyanin synthesis by FR. (Received March 14, 1988; Accepted September 28, 1988)     

Early-age-dependent selective decrease of differentiation potential of hair-follicle-associated pluripotent (HAP) stem cells to beating cardiac-muscle cells

We have previously discovered nestin-expressing hair-follicle-associated pluripotent (HAP) stem cells and have shown that they can differentiate to neurons, glia, and many other cell types. HAP stem cells can be used for nerve and spinal cord repair. We have recently shown the HAP stem cells can differentiate to beating heart-muscle cells and tissue sheets of beating heart-muscle cells. In the present study, we determined the efficiency of HAP stem cells from mouse vibrissa hair follicles of various ages to differentiate to beating heart-muscle cells. We observed that the whiskers located near the ear were more efficient to differentiate to cardiac-muscle cells compared to whiskers located near the nose. Differentiation to cardiac-muscle cells from HAP stem cells in cultured whiskers in 4-week-old mice was significantly greater than in 10-, 20-, and 40-week-old mice. There was a strong decrease in differentiation potential of HAP stem cells to cardiac-muscle cells by 10 weeks of age. In contrast, the differentiation potential of HAP stem cells to other cell types did not decrease with age. The possibility of rejuvenation of HAP stem cells to differentiate at high efficiency to cardiac-muscle cells is discussed.