RADIOAUTOGRAPHIC STUDY OF IN VIVO AND IN VITRO INCORPORATION OF FUCOSE-3H INTO THYROGLOBULIN BY RAT THYROID FOLLICULAR CELLS

The incorporation of fucose-³H in rat thyroid follicles was studied by radioautography in the light and electron microscopes to determine the site of fucose incorporation into the carbohydrate side chains of thyroglobulin, and to follow the migration of thyroglobulin once it had been labeled with fucose-³H. Radioautographs were examined quantitatively in vivo at several times after injection of fucose-³H into rats, and in vitro following pulse-labeling of thyroid lobes in medium containing fucose-³H. At 3–5 min following fucose-³H administration in vivo, 85% of the silver grains were localized over the Golgi apparatus of thyroid follicular cells. By 20 min, silver grains appeared over apical vesicles, and by 1 hr over the colloid. At 4 hr, nearly all of the silver grains had migrated out of the cells into the colloid. Analysis of the changes in concentration of label with time showed that radioactivity over the Golgi apparatus increased for about 20 min and then decreased, while that over apical vesicles increased to reach a maximum at 35 min. Later, the concentration of label over the apical vesicles decreased, while that over the colloid increased. Similar results were obtained in vitro. It is concluded that fucose, which is located at the end of some of the carbohydrate side chains, is incorporated into thyroglobulin within the Golgi apparatus of thyroid follicular cells, thereby indicating that some of these side chains are completed there. Furthermore, the kinetic analysis demonstrates that apical vesicles are the secretion granules which transport thyroglobulin from the Golgi apparatus to the apex of the cell and release it into the colloid.     

Mirror responses in a Japanese macaque troop (Arashiyama West)

Behavior toward two mirrors in the field was observed in the Arashiyama West troop ofMacaca fuscata. Counts of visits to the mirrors, plus scan and focal animal sampling were used. Some animals were marked with fluorescent paint to test informally for self-recognition. A relatively high mean frequency of visits to the reflecting side of both mirrors by all age classes, ranks, and sexes was recorded. There was no age difference in frequency of mirror visits per sample but adults spent more time per visit than subadults who in turn spent more time than juveniles. There was no indication of self-recognition by paint-marked animals. Mirrors appeared to be used to monitor the reflected scene and to look at the self-image. Social behavior in the mirror zone that was not directed toward the mirror was common to all age classes. Species-typical behavior directed toward the mirror was seen in younger animals but very seldom in adults. No threat displays by any animal were observed. We suggest that for adults the mirror image was not seen simply as another monkey.     

Neurochemical Abnormalities in Brains of Renal Failure Patients Treated by Repeated Hemodialysis

We examined autopsied brain from 10 patients with end-stage renal failure who had undergone repeated hemodialysis. Eight had classic symptoms, and two had suggestive symptoms of dialysis encephalopathy. Findings were compared with those in autopsied brain from control adults who had never been hemodialyzed. Mean gamma-aminobutyric acid (GABA) contents were significantly reduced in frontal and occipital cortex, cerebellar cortex, dentate nucleus, caudate nucleus, and medial-dorsal thalamus of the hemodialyzed patients, the reduction being greater than 40% in cerebral cortex and thalamus. Choline acetyltransferase activity was reduced by 25-35% in three cortical regions in the hemodialyzed patients. These two abnormalities were observed in the brain of each hemodialyzed patient, regardless of whether or not the patient died with unequivocal dialysis encephalopathy. Pyridoxal phosphate contents were substantially reduced in brains of the hemodialyzed patients, but metabolites of noradrenaline, 3,4-dihydroxyphenylethylamine (dopamine), and 5-hydroxytryptamine (serotonin) were present in normal amounts. Aluminum levels were abnormally high in frontal cortical gray matter in the hemodialyzed patients. Although this study does not clarify the role played by aluminum toxicity in the pathogenesis of dialysis encephalopathy, the abnormalities we found suggest the need for further neurochemical investigations in this disorder.     

An improved polyurethane support system for monitoring growth in plant cell cultures

An improved culture system for plant cells that employs filter paper resting on polyurethane saturated with liquid medium is described. It combines a simplified version of the system outlined by Weber and Lark [1979, Theor Appl Genet 55: 81–86] with the method of growth estimation described by Horsch et al. [1980, Can J Bot 58: 2402–2406]. The growth of plated cells or callus can be conveniently monitored through repeated non-destructive fresh weight measurements of the filter paper and adhering cells, thereby allowing the construction of a complete growth curve over the course of an experiment. Experiments with 3 Nicotiana genotypes (N. plumbaginifolia Viv., N. tabacum L. SC 58 and N. tabacum WI 38) and 3 Vitis vinifera L. genotypes (Chenin Blanc, Dogridge and White Riesling) clearly demonstrate higher growth rates of plated cells on polyurethane supports compared with agar. Further experiments with N. plumbaginifolia illustrate the use of polyurethane supports for culturing cells at low pH (4.0) and the recovery of spent medium for monitoring changes in pH. These features will greatly facilitate quantitative studies of mineral nutrition and metal toxicity in cultured cells. Polyurethane supports also allow the incorporation of conditioned medium or feeder cells to support the growth of cells at low densities and facilitate the rapid recovery of variant cells.     

A method for probing the affinity of peptides for amphiphilic surfaces

We have developed a rapid method for probing the affinity of peptides toward an amphiphilic surface. Hydrophobic polystyrene-divinylbenzene beads of 5.7 +/- 1.5 micron diameter are coated with a monomolecular film of egg lecithin to achieve the equilibrium spreading density of the phospholipid, 6 X 10(-3) molecule/A2. The coated beads are ideally suited for assessing the affinity of peptides for phospholipid surfaces: Large quantities of lipid-coated beads of known surface area can be prepared easily and rapidly. Within the pH range 2.0 to 9.0, the adsorbed phospholipids are relatively resistant to hydrolysis and remain bound indefinitely. Following incubation with peptide ligands, beads can be separated from the reaction mixture by centrifugation. Peptides, such as melittin, which destroy or cause fusion of single bilayer phospholipid vesicles, cannot disrupt lecithin-coated beads in a comparable way, and do not displace lecithin from the surface of beads. After incubating these beads in solutions of peptides and proteins, we have determined the parameters for the binding of several ligands to the phospholipid surface. The binding of many amphiphilic peptides obeys a Langmuir adsorption isotherm, i.e., saturable reversible binding to independent and equivalent sites on the bead. That the binding is a true reversible equilibrium is shown by desorption of the ligand upon dilution. From the isotherm, the surface areas occupied by the ligand molecules were calculated, and were observed to be similar to those observed in monolayers at the air-water interface. In comparing the binding of amphiphilic peptides to that of completely hydrophilic peptides, we observed that only the former bind at levels measurable by our techniques. Thus, this method can serve as a rapid assay for detecting amphiphilicity in peptides of putative amphiphilic character.     

Role of decreased numbers of follicles on reproductive performance in young and aged rats

The primary objectives of this study were to: 1) determine if removal of 1.5 ovaries from young rats would mimic reproductive characteristics that normally occur with advancing age and 2) determine if removal of 1.5 ovaries from aged rats would further advance the process of reproductive aging. Removal of 1.5 ovaries increased the number of young (P less than 0.05) and old (P less than 0.01) rats that exhibited abnormal estrous cycles. In addition, concentrations of follicle-stimulating hormone (FSH) were higher at both ages in the groups with half an ovary. The increased concentrations of FSH are consistent with a decrease in the number of growing follicles after removal of 1.5 ovaries. All groups had lower concentrations of estradiol (E2) than young controls. There was a significant increase in the number of abnormal embryos with age and removal of 1.5 ovaries when rats were mated during a 5-day estrous cycle, but there was no effect if they were mated during a 4-day estrous cycle. From the results of this study, we conclude that the reduction in ovarian tissue in young and aged rats mimicked several reproductive characteristics of advancing age. Also, an effect of aging on the hypothalamus was evident in this study.     

Geographic parthenogenesis in the stream mayfly Eurylophella funeralis in eastern North America

Unisexual populations were found largely at the periphery of the geographic range of the mayfly Eurylophella funeralis in eastern North America. Bisexual populations generally had a normal sex ratio but at least two populations were observed with only about 2–11% males. Hatch success for unfertilized eggs depended on temperature but in general averaged >61% in unisexual populations and < 14% in bisexual populations. Eggs took about 65 d to hatch at 10°C and 19 d at 25°C; no hatching occurred at 5 or 30°C. Successful oviposition was observed for about 97% and 20% of unmated female adults from unisexual and bisexual populations respectively. The hatch success of unfertilized eggs for three geographically distinct bisexual populations was in versely correlated with the proportion of males in the populations. The genetic structure of bisexual and unisexual populations was very similar. Unisexual populations consists of clones, with as few as four recognizable phenotypes being present at a given location. All offspring had the same phenotype as the mother. Tychoparthenogenesis is suggested as the most reasonable working hypothesis concerning the origin of unisexual populations in E. funeralis .     

Eccentric exercise-induced muscle damage impairs muscle glycogen repletion

Five healthy untrained young male subjects were studied before, immediately after, and 10 days after a 45-min bout of eccentric exercise on a cycle ergometer (201 W). The subjects were sedentary at all other times and consumed a eucaloric meat-free diet. Needle biopsies of the vastus lateralis muscle were examined for intracellular damage and glycogen content. Immediately after exercise, muscle samples showed myofibrillar tearing and edema. At 10 days, there was myofibrillar necrosis, inflammatory cell infiltration, and no evidence of myofibrillar regeneration. Glycogen utilization during the exercise bout was 33 mmol glycosyl units/kg muscle, consistent with the metabolic intensity of 44% of maximal O2 uptake; however, the significant glycogen use by type II fibers contrasted with concentric exercise performed at this intensity. At 10 days after exercise, muscle glycogen was still depleted, in both type I and II fibers. It is possible that the alterations in muscle ultrastructures were related to the lack of repletion of muscle glycogen. Damage produced by eccentric exercise was more persistent than previously reported, indicating that more than 10 days may be necessary for recovery of muscle ultrastructure and carbohydrate reserves.     

ULTRASTRUCTURE OF NOCTILUCA MILIARIS (PYRROPHYTA) WITH GREEN FLAGELLATE SYMBIONTS1,2

The ultrastructure of the Noctiluca miliaris Suriray from Southest Asian waters which contains the green flagellate, Pedinomonas noctilucae (Subrahmanyan) Sweeney, is in all major respects similar to that of the European strain. New details of the thecal vesicles, pellicle and underlying microtubules are presented. The possibility that the lipid vesicles are identical with the strongly phase-retarding bodies in the surface cytoplasm, some of which are “microsources” of bioluminescence, is suggested.