Health beliefs of hypertensive patients in a family medicine residency program. This paper presents the results of a survey of the hypertensive patient population in a university family medicine practice to determine health beliefs, patient perceptions of the severity of their condition, stated levels of compliance to drug regimens, frequency of drug side effects, and frequency of physician discussions of drug side effects and diet. Health beliefs were then associated with diastolic hypertension and increase in medication dosage over a four-month period. Results of the survey sharpen the focus of patient education efforts by the family physician on specific attitudes, beliefs, and treatment issues which are most appropriate for the hypertensive patient. The physician may improve his/her management of hypertensive patients by: (1) anticipating prevalent myths and misconceptions which patients have concerning hypertension; (2) alleviating patient anxiety by stressing that control of blood pressure decreases the likelihood of complications; (3) effectively communicating to each patient the current status of his/her condition at each visit; and (4) initiating discussion of side effects through direct questioning for those effects most frequently seen.


The after-hours call in family practice. Patient care after hours continues to be an important part of the work of family physicians, in spite of the trend towards increasing Emergency Room utilization. In this paper the literature concerning after-hours care in family practice is reviewed in terms of definition, demography, utilization, morbidity, and patient stereotypes. In the Family Practice Residency Program of the University of North Carolina, 4,760 after-hours calls were recorded over two years by residents and faculty physicians. Seventy-two percent of the calls were handled purely on the telephone with little variation for patient age groups. The overall call rate was 474 calls per 1,000 patients per year. Fever and skin wounds were the most frequent symptoms recorded and respiratory tract infections, minor trauma, and anxiety were the commonest diagnoses. Thirteen percent of the contacts engendered anger or frustration in the physician. A survey of patients calling after hours demonstrated a lack of congruence between physician and patient concerning the main reason for the call in over 30 percent of contacts.


Epigenetic formation of lactate dehydrogenase isozymes in the house mouse, Mus musculus. The origin of mouse lactate dehydrogenase (LDH) sub-bands was investigated by using our miniaturized polyacrylamide gel electrophoretic apparatus. Mouse LDH isozymes are generated by combinations of three types of A subunit, the primary type and two epigenetically modified forms. These are designated A1, A2, and A3 in the order of their electrophoretic mobilities towards the anode. The A1 subunit arises from the covalent binding of molecules of glutathione through disulfide bonds to the original subunit, A3. The A2 subunit arises from the covalent binding of molecules of cysteine through disulfide bonds to the A3 subunit. All isozymes can be explained as tetramers composed of the three kinds of A subunit (A1, A2, or A3) in combination with B subunits to yield a total of 35 isozymes. The kinetic properties of these sub-bands were also examined. There was no difference between A24 and A34 in the Km for pyruvate and for lactate. Thermostability at 56 degrees C was greater for A34 than for A24. The activities of tetramers at the electrophoretic position of A3B1 and A4 in extracts containing all five isozymes were increased by treatment of the extracts with high concentrations of reduced glutathione or cysteine with the concomitant disappearance or decrease in activity of tetramers at the position of B4 and A3B1. These results suggest that, in the presence of reduced glutathione or cysteine, LDH isozymes containing the B subunit are first dissociated and then the A subunits are preferentaially recombined.


The distribution of sorbitol dehydrogenase in embryonic and adult chicken tissues. Sorbitol dehydrogenase activity (SDH) has been determined in various organs of embryonic and adult chickens. SDH is present in 24-hour embryos, and its activity continues to rise during the next 48 hours. During embryonic development and after hatching, regional differences in SDH activity are demonstrable in the organs of the animal. These differences concern both level of enzyme activity and temporal changes with development. No correlation could be established between enzymatic activity and the fructose concentration of the organs studied.


Light-induced acceleration of retina development in a mouth-brooding teleost. Rearing animals in the dark had been shown to be either without effect on the development of the retina or to result in a reduction or a delay of retinal maturation. In the present study, the influence of light on a retina which normally develops under conditions of very dim light has been investigated. When 3-day-old embryos of the mouth-brooding teleost Tilapia leucosticta are placed into a continuously lighted environment, from day 6 on, optic tract diameter and inner plexiform layer thickness increase up to day 10 or 12. In the dark-reared retina, this increase occurs only after day 10. Similarly, synaptic junctions of the inner plexiform layer appear at about day 6 in the light-reared retina and increase continuously on following days, whereas in the retina of embryos reared in darkness, they appear at day 7 and do not increase essentially before day 11. These and other data suggest that continuous light induces a precocious growth of retinal structures. The first differences between light- and dark-reared retinae appear synchronously with the beginning of receptor cell development and prior to the first synaptic junctions. A non-neurally mediated effect of light on the retinal ganglion cells is consequently assumed.


Interactions between irradiated and unirradiated tissues during supernumerary limb formation in the newt. The interactions between irradiated and unirradiated blastemas and stumps in the newt forelimb were studied. Irradiated right blastemas at the stage of early digits were grafted to unirradiated left stumps and unirradiated left blastemas were grafted to irradiated right stumps. Grafts were oriented with their anterior-posterior axes opposed to that of the stumps. Supernumerary limbs ranging in completeness from one to four digits were found to arise predominantly on the anterior or posterior sides of the host limb. The graft developed well when the blastema was unirradiated and had reversed handedness with respect to the stump. Irradiated grafts developed poorly. On occasions, limbs with two supernumerary structures were found. The results are discussed in terms of the origin of the cells which comprise the supernumerary limbs and their bearing on a recently presented model concerned with pattern specification and regulation in epimorphic fields.


Periodic breathing in the crocodile, Crocodylus niloticus: consequences for the gas exchange ratio and control of breathing. The ventilatory pattern in the Nile crocodile consists of episodes of breathing, interrupted by periods of breath-holding, the latter occupying 80% of total time during normal breathing at 25 degrees C. End-tidal gas composition varied with the periodic breathing but PO2 was always high (PO2 less than 110 torr) and PCO2 low (PCO2 less than 25 torr). The alveolar gas exchange ratio, RE, was very low during the non-ventilatory periods (RE congruent to 0.5), but increased markedly during ventilation. Breathing of hypoxic and hpercapnic gases caused a pronounced decrease in the duration of breath-holds. Hypercapnia decreased breathing frequency during ventilatory periods, but increased tidal volume. The results are discussed in relation to the practice of prolonged breath-holding associated with diving in crocodiles.


DNA polymerase activity in preimplantation mouse embryos. DNA polymerase activity was measured in mouse embryos at stages before implantation to determine whether it increases in proportion to the amount of DNA synthesis, as it does in populations of differentiated mammalian cells, or remains constant, as it does in early sea urchin embryos. Total enzyme activity was found to be relatively unchanged following fertilization and in the first few cleavage stages. However, between the 12- and 120-cell (blastocyst) stage, the amount of activity increased by several-fold. These results indicate that the relationship between amount of DNA polymerase activity and DNA synthesis in mouse embryos exhibits two phases: in the early cleavage phase it is similar to that in sea urchin embryos, whereas, in the blastocyst phase, it is similar to that in differentiated mammalian cells.


DNA reassociation kinetics in diploid and phylogenetically tetraploid cyprinidae. Four diploid and three phylogenetically tetraploid Cyprinidae (Ostariophysi) have been characterized as for nuclear DNA content, modal chromosome number and DNA reassociation kinetics (hydroxyapatite chromatography). Among the diploid species nuclear DNA content (10(-12) g DNA/2C) was 1.62 for Tinca tinca, 1.87 for Scardinius erythrophthalmus, 2.53 for Leuciscus cephalus and 2.75 for Alburnus alburnus, while the phylogenetically tetraploid species Carassius auratus, Barbus barbus and Cyprinus carpio attained 3.40, 3.66 and 3.80 respectively. Modal chromosome number was 2n = 48-50 for diploid individuals and 2n = 100-104 for phylogenetically tetraploid ones. In all the species 5--8% of the genome is represented by highly repetitive and foldback DNA. In DNA reassociation kinetics of phylogenetically tetraploid Cyprinidae a distinct plateau separates an intermediate reassociating sequence fraction (about 22% of the genome; with average repetition frequencies between 1,000 and 1,400) from a slow reassociating one (unique DNA; about 72% of the genome). These two genome fractions are not clearly distinguishable from each other in Cot curves of the diploid Cyprinidae, where a similar plateau is not evident. Since simple ploidy changes are not expected to affect DNA reassociation kinetics we suggest a different evolution in the genome organization of the two ploidy groups. Some possible hypotheses are discussed.


Fatty acid and sterol synthesis by hepatocytes of thermally acclimated rainbow trout (Salmo gairdneri). Incorporation of tritium from tritiated water into lipid fractions was measured in isolated hepatocytes from rainbow trout (Salmo gairdneri) acclimated to 5 degrees C and 20 degrees C. Hepatocytes from cold-acclimated trout exhibited significantly higher rates of tritium incorporation into both fatty acid and sterol fractions at assay temperatures of 15 degrees C and 20 degrees C than did hepatocytes from warm-acclimated trout. Tritium incorporation into the fatty acid fraction was nearly temperature independent in hepatocytes from warm-acclimated trout (Q10 = 1.39) but markedly temperature dependent (Q10 = 2.63) in hepatocytes from cold-acclimated trout; in contrast, rates of sterol synthesis were more temperature dependent in warm-acclimated trout. At 5 degrees C, fatty acid lipogenesis comprised a significantly greater percentage of the total tritium incorporation in hepatocytes from warm-acclimated trout and the percentage of total lipogenesis attributable to fatty acids decreased significantly in warm-acclimated trout as the assay temperature increased; the opposite trends were observed in cold-acclimated trout.


Neurites grow faster towards the cathode than the anode in a steady field. We explanted fragments of embryonic chick dorsal root ganglia on to polylysine coated glass and cultured them in a medium containing one unit of nerve growth factor plus enough methylcellulose to give viscosities from 0.01-3,000 poise. We allowed them to grow out in the absence of a field, and then selected explants with halos of neurites which were relatively dense, relatively symmetrical, and practically free of glial cells. These selected explants were then exposed to electrical fields of up to 140 mV/mm for some hours. In media with viscosities of one poise or less, the field some times dragged the central cell mass of an explant towards the anode. However, in cases where the central cell mass did not move, fields of 70-140 mV/mm induced that sector of each neurite halo which faced the cathode to grow out several times faster than the one facing the anode.


Inhibition of desmosome formation in chick cell aggregates. Desmosomes (macula adherens) have been associated with the function of adhesion. Their possible role in aggregation and sorting of chick and mouse epithelial cells has been investigated. Treatment of aggregates with 2-5 microgram/ml of actinomycin D which inhibited RNA synthesis also inhibited both desmosome formation and aggregation if administered at the beginning of the aggregation process. In contrast, if the drug was administered at six hours, when the cells had recovered from the process of dissociation, then aggregation over the following six hours appeared normal from observation of living samples. Such aggregates incorporated leucine-3H at roughly 85% of the control level. A quantitative comparison was made of desmosome formation in aggregates treated with actinomycin D for hours 6-12 and those cultured in normal medium. Desmosome formation was inhibited by the drug, although aggregation could proceed. Combinations of chick corneal and mouse skin cells sorted out in the presence of actinomycin D to the same extent as controls. Thus desmosome formation, which normally occurs during aggregation of the epithelial cells studied here, is not coupled with the aggregation or cell sorting process in these cells of stratified epithelia. When cells were treated with cycloheximide (100 muM) both desmosome formation and the progressive rounding up of aggregates was inhibited.


Acrosin does not appear to be bound to the inner acrosomal membrane of bull spermatozoa. Ferritin-conjugated soybean trypsin inhibitor was used for the ultrastructural localization of acrosin in bull spermatozoa following acrosomal disruption. The ferritin label was observed in the anterior segment of the acrosome in disrupted cells only. Emptied acrosomes were labeled, mostly on the external surface of their outer membrane. Labeling was also found on the material bound to detached acrosomal caps. However, at no time could the ferritin label be found on the inner acrosomal membrane. It is concluded that acrosin activity is not present on the inner acrosomal membrane but is lost from the acrosomal matrix as the acrosomal reaction proceeds.


Delayed reproductive regression in male hamsters bearing intrarenal pituitary homografts and kept under natural winter photoperiods. The testes and accessory sex organs of adult male hamsters regressed to an infantile state when the animals were maintained under natural photoperiodic and temperature conditions beginning November 13. The atrophy was evident after both five and ten weeks exposure to these conditions. Either one or two prolactin-secreting pituitary grafts placed under the kidney capsule greatly delayed the testicular and accessory organ regression. Animals bearing pituitary grafts also had higher circulating levels of immunoreactive prolactin while plasma luteinizing hormone values were similar in all groups of hamsters.


Enzyme changes accompanying thermal acclimation in two species of pleurocerid snails. Goniobasis cahawbensis is a stream snail that experiences an annual temperature cycle. G. cochliaris is limited in distribution to springs, and their immediate vicinities, which are characterized by nearly constant annual temperatures. The present study sought to determine whether temperature dependent biochemical differences exist that might account for the differential distribution of these congeneric pleurocerid snails. Eight enzymes were examined following acclimation to 10 degrees, 17 degrees and 24 degrees C. No significant temperature dependent qualitative differences in enzyme phenotypes were demonstrable in either species by starch-gel electrophoresis for malate dehydrogenase, glucose-6-phosphate dehydrogenase, phosphogluconate dehydrogenase, phosphoglucomutase, superoxide dismutase and acetyl and butyryl esterases. Significant quantitative differences were observed in three of these enzymes. G. cahawbensis cytosol malate dehydrogenase activity increased significantly with increasing acclimation temperature, while G. cochliaris malate dehydrogenase activity remained unchanged. The activities of glucose-6-phosphate dehydrogenase did not differ significantly between acclimation temperatures for either species; however, the overall activity of both enzymes was significantly higher for G. cochliaris. Appreciable levels of LDH activity were not demonstrable by electrophoresis or enzymatic assay.


Specificity of inhibition by steroids of porcine oocyte maturation in vitro. In order to examine the influence of several steroids on the process of oocyte maturation, denuded (adherent cumulus granulosa cells mechanically removed) and intact (cumulus granulosa cells left attached) porcine oocytes were cultured in the presence or absence of estradiol-17 beta, estradiol-17 alpha, testosterone, cortisol, progesterone, or the nonsteroidal estrogen diethyl stilbestrol (all at 10 microgram/ml) in defined medium that contained either BSA or dextran. Estradiol-17 beta was the only steroid to exert a significant inhibitory effect on the maturation of denuded oocytes, and did so only in BSA supplemented medium. The inhibition was reversible in that oocytes, cultured in steroid-free medium after initial culture in estradiol-17 beta medium, resumed meiotic maturation. Oocytes took up 3H-estradiol-17 beta in both media, although less radiolabel entered oocytes in BSA supplemented medium. The majority of label in the oocytes, when cultured with either medium, was not displaced by excess radioinert estradiol-17 beta or progesterone, nor were the oocytes saturated even when cultured in 10(-6) M estradiol-17 beta. Autoradiography of sectioned oocytes after culture in 3H-estradiol-17 beta has shown that there was no selective accumulation of silver grains over the germinal vesicle as was the case with granulosa cell nuclei. This observation suggests that estradiol-17 beta may not act at the level of the oocyte nucleus.


Trochlear-oculomotor nerve interactions in Xenopus laevis tadpoles: a temporal study. When the trochlear nerve (NIV), which innervates the superior oblique muscle (SOM), is crushed or cut at stages 48-49 in Xenopus tadpoles, fibers from the oculomotor nerve (NIII) sprout and invade the SOM. The maximal percentage of specimens having at least one oculomotor nerve fiber on the SOM on a given day increased from 9.1% following a single crushing of NIV to 84.2% following three successive severings of NIV and the average number of silver-impregnated NIII fibers per specimen increased from 0.23 +/- 0.16 (mean +/- S.E.M.) in the single-crush experiment to 7.35 +/- 1.33 in the triple-cut experiment. This increase directly reflects the delay in the return of NIV. As NIV returns to the SOM, a portion of the inappropriate innervation is lost; while another portion appears to be stable and is in evidence 90 days after a single sectioning of NIV. The more rapidly NIV returns to the SOM, the more complete is the displacement of the NIII fibers. This suggests that the association between NIII and the SOM changes with time so that easy displacement of the inappropriate innervation is likely only when the reinnervation by the appropriate nerve fibers is rapid.


Reduction of sodium dependent stump currents disturbs urodele limb regeneration. We have asked the question whether the natural electric currents which leave urodele limb stumps are in any way needed for their regeneration. As an initial test, we have greatly reduced such currents in the tiger salamander, Ambystoma tigrinum, by applying 0.5 mM amiloride to the stump skin or by immersion of the animals in sodium depleted media. We have also reduced such currents in the red spotted newt, Notophthalmus viridescens, by such immersion. Limb regeneration in half of the amiloride-treated animals was either entirely blocked or grossly deficient, while the others regenerated normally. Limb regeneration in sodium depleted media was consistently inhibited for some weeks but then recovered. These results are consistent with the hypothesis that stump currents are in some way needed for normal regeneration.


Stages of forelimb regeneration in Ambystoma maculatum. A series of normal stages describing the regeneration of larval A. maculatum limbs after amputation through the upper arm or wrist is described. Nine discrete stages were recognized, based on external morphological and associated histological features. These stages are Initial Dedifferentiation (ID), Early Bud (EB), Medium Bud (MB), Late Bud (LB), Early Redifferentiation (ER), Notch (N), 2-Fingerbud (2-FB), 3-Fingerbud (3-FB) and 4-Fingerbud (4-FB). Similarities and differences between this and other staging systems for urodele limb regeneration are discussed. The absence of osteoclasts was a striking feature during dedifferentiation of the wrist, in contrast to their presence in large numbers during dedifferentiation of the upper arm.


Reproductive failure and maternal-fetal relationship in a Peromyscus species cross. Mating, fertilization, implantation, prenatal mortality, fetal and placental size, and placental ultrastructure were studied in intraspecific and interspecific crosses involving Peromyscus maniculatus and P. polionotus. Failure to mate was a major factor in interspecific crosses and was much more pronounced in crosses between P. polionotus females and P. maniculatus males than in the reciprocal cross. Failure of implantation following mating, however, was more pronounced in crosses between P. maniculatus females and P. polionotus males. Failure of implanted embryos to survive to term was a factor in crosses between P. polionotus females and P. maniculatus males. Comparison of the placental labyrinth of conceptuses from intraspecific and interspecific crosses revealed no differences at the ultrastructural level. The relationship of these observations to the evolution of isolating mechanisms in mammals and to physiological aspects of the developing maternal-fetal relationship are discussed. A model of placental and fetal size inheritance is presented.


Limbless: a new genetic mutant in the chick. A mutation in the chicken resulting in total amelia is described. Genetic analysis indicates that the limbless condition is due to an autosomal recessive gene. In the limbless embryos the apical ectodermal ridge is lacking. However both the pectoral and pelvic girdles as well as the respiratory and excretory systems are normal. In the affected embryos usually the upper beak is shorter than the lower beak.


Separation of the epithelial and mesenchymal components of the newt limb regenerate with salt. After incubation of isolated forelimb regenerates of Notophthalmus (Triturus) viridescens at all developmental stages for 60 minutes at 37 degrees C in a salt medium containing 111 mM sodium chloride, 5.6 mM potassium chloride and 100 mM sodium phosphate buffer at pH 7.5, the wound epithelium of each regenerate was removed intact from its underlying mesenchymal component. The suggestion is made that the salt medium is an effective epithelial-mesenchymal separating agent due to a combination of its hypertonicity, high ionic strength and the fact that the medium precipitates calcium as calcium phosphate. Attempts to dissect away the epithelium from the mesenchyme after incubation of isolated regenerates in sodium phosphate containing 1% or 3% Difco 1:250 trypsin, 10 mM EDTA or 150 units collagenase/ml medium were unsuccessful. Epidermis of adult newt forelimb skin was removed only after extended incubation of the forelimbs in the salt medium for three hours at 37 degrees C or after freezing isolated forelimbs in buffer and subsequent thawing.


Allopurinol-induced melanism in the tiger salamander (Ambystoma tigrinum nebulosum). The enzyme, xanthine dehydrogenase (XDH), has been examined in Ambystoma tigrinum nebulosum with respect to its role in pigmentation. It now seems probable that the melanoid gene (m) either codes directly for XDH or is somehow intimately connected with the normal function of this enzyme. Inhibition of XDH using the drug, allopurinol, results in animals which appear to be phenocopies of melanoid mutants as described for the Mexican axolotl (Ambystoma mexicanum). The effects of allopurinol in terms of specific pigmentary alterations were examined, and a new method for analyzing heterogeneous extracts of skin pigments (e.g., purines and pteridines) is presented. The significance of the link between XDH and melanism is discussed with emphasis on possible mechanisms of pigment induction and general applicability to biological systems.


Liquid chromatographic demonstration of catecholamine release in fish heart. High-pressure liquid chromatography (HPLC) was used to assay for catecholamines released into the heart of the goldfish (Carassius auratus L.) upon stimulation of vagal nerve fibers. Stimulation induced the release of norepinephrine, epinephrine and dopamine into the lumen of the perfused, isolated heart. The results suggest that heart activity in teleosts is directly modulated by functional adrenergic, cardioacceleratory nerve fibers.


Ionic and osmotic equilibria of human red blood cells treated with nystatin. Human red blood cells have been incubated in the presence of nystatin, which allows Na and K, as well as Cl and pH to equilibrate rapidly when cell volume is set with external impermeant sucrose. The intracellular mean ionic activity coefficients, relative to values in the extracellular solution, for KCl and NaCl are 1.01 +/- 0.02 and 0.99 +/- 0.02 (SD, n = 10), respectively, and are independent of external pH, pH o, and of [sucrose]o. With nystatin the dependence of red cell volume on [sucrose]o deviates from ideal osmotic behavior by as much as a factor of three. A virial equation for the osmotic coefficient, phi, of human hemoglobin, Hb, accounts for the cell volumes, and is the same as that which describes Adair's measurements of phi Hb for Hb isolated from sheep and ox bloods. In the presence of nystatin the slope of the acid-base titration curve of the cells is independent of cell volume, implying that the charge on impermeant cellular solutes is independent of Hb concentration at constant pH. By modifying the Jacobs-stewart equations (1947. J. Cell. Comp. Physiol. 30: 79--103) with the osmotic coefficients of Hb and of salts, a nonideal thermodynamic model has been devised which predicts equilibrium Donnan ratios and red cell volume from the composition of the extracellular solution and from certain parameters of the cells. In addition to accounting for the dependence of cell volume on osmotic pressure, the model also describes accurately the dependence of Donnan ratios and cell volumes on pHo either in the presence or absence of nystatin.


Nonparallel isometric tension response of rabbit soleus skinned muscle fibers to magnesium adenosine triphosphate and magnesium inosine triphosphate. The isometric tension response of single "skinned' rabbit soleus muscle fibers to MgATP and McITP in the absence of calcium was studied. [MgATP] or [MgITP] was varied in solutions of ionic strength 0.30 and temperature 20 degrees C. Steady-state tension that developed in MgATP or MgITP solutions was a biphasic bell-shaped function of log [MgATP] or log [MgITP] which increased from zero to maximum tension and then declined again to zero. Analysis of the data showed that, under comparable ionic conditions, percent tension vs. log [MgATP] and percent tension vs. log [MgITP] curves are not parallel. Instead, the percent tension vs. log [MgITP] curve is much broader. Additionally, under comparable ionic conditions maximum tension in MgITP solutions was higher than in MgATP solutions. In addition, in MgATP solutions, pH, [K+], and excess ATP were varied. Raising pH from 7 to 8, [K+] from 46 mM to 200 mM, or decreasing excess ATP from 2 to 0.5 mM all increased maximum tension. None of these factors, however, influenced the shape or position of the percent tension vs. log [MgATP] curve.


Receptive field mechanisms of cat X and Y retinal ganglion cells. We investigated receptive field properties of cat retinal ganglion cells with visual stimuli which were sinusoidal spatial gratings amplitude modulated in time by a sum of sinusoids. Neural responses were analyzed into the Fourier components at the input frequencies and the components at sum and difference frequencies. The first-order frequency response of X cells had a marked spatial phase and spatial frequency dependence which could be explained in terms of linear interactions between center and surround mechanisms in the receptive field. The second-order frequency response of X cells was much smaller than the first-order frequency response at all spatial frequencies. The spatial phase and spatial frequency dependence of the first-order frequency response in Y cells in some ways resembled that of X cells. However, the Y first-order response declined to zero at a much lower spatial frequency than in X cells. Furthermore, the second-order frequency response was larger in Y cells; the second-order frequency components became the dominant part of the response for patterns of high spatial frequency. This implies that the receptive field center and surround mechanisms are physiologically quite different in Y cells from those in X cells, and that the Y cells also receive excitatory drive from an additional nonlinear receptive field mechanism.


Isolation and characterisation of a defective measles virus from a subacute sclerosing panencephalitis patient. A cytopathic measles virus was isolated from a brain biopsy of a subacute sclerosing panencephalitis (SSPE) patient. The agent could be transferred to Vero cells by cocultivation, but the infectivity always remained cell-associated -ie, a defective virus infection. The cell-associated nature of the virus was retained through 25 passages in Vero cells. Intracerebral inoculation of hamsters (2-6 days old) with the cocultured Vero cells gave rise to 100% mortality in 5-7 days. The virus retained its cell-associated nature after passage in hamsters. Electron microscopy of the brain and Vero cocultures showed the presence of virus-like ribonucleoparticles mainly in the nucleus. The presence of viral antigens in the nucleus, cytoplasm, and on the plasma membranes was confirmed by immunofluorescence. Using a combination of immunological and biochemical techniques, it was shown that all the viral proteins were synthesized with the exception of the haemagglutinin. Inclusion of the fusion inhibitor SV4814 (CBZ-D phenylalanine-L-phenylalanine-L-arginine-NO2) in the culture medium led to the elimination of the SSPE infection.


Deeper insight into HBsAg--anti-albumin antibody correlations. The presence and titers of anti-albumin antibodies (AAA)--ie, precipitins (AA-IP) and agglutinins (AA-Aggl)--and the serum concentrations of immunoglobulins and albumin were determined in 210 asymptomatic carriers of HBsAg grouped according to HBsAg titer and subtype. A different immunologic pattern was observed in the HBsAg/ad and HBsAg/ay carriers, the results suggesting an increased aggressivity for the HBsAg/ay subtype which was evident in subjects with lower HBs antigenemia and characterized by higher concentrations of IgG and IgM and rises in AA-P titers. A positive linear correlation was found between HBsAg and AA-Aggl titers; the carriage of HBsAg/ad was associated with significant higher values of AA-Aggl than of HBsAg/ay. These subtle correlations between HBsAg titer and subtype and AAA suggest that the human serum albumin (HSA) known to exist on the HBsAg particles may be similar in a modified form, to that representing AAA specificity.


Reaction time crossover in process schizophrenic patients, their relatives, and control subjects. Of 70 process schizophrenic patients clinically assessed and tested for reaction time, 40 showed reaction time crossover. Of the 40 with crossover, 53 of their first degree relatives were clinically assessed and tested. Five of the relatives, found to have psychotic symptoms, were analyzed separately. Also, 41 control subjects without personal or family history of psychiatric disturbance were clinically assessed and tested. The nondisturbed relatives had reaction time crossover (regular minus irregular) significantly greater than the (expected) negative values of the normal control subjects, but not significantly greater than a zero value within subjects. When disturbed and nondisturbed relatives were combined, greater differences indicative of crossover occurred. On the other hand, the relatives, as expected, did not show the slow mean reaction time characteristic of diagnosed schizophrenic patients.


On the meaning of electrodermal nonresponding in schizophrenia. The skin conductance responses of schizophrenics, neurotics and normals to orienting stimuli were examined. Only the schizophrenic group showed a high incidence of absence of skin conductance orienting responses. A subdivision of the schizophrenic sample into thosee who did not show any orienting response and those who responded more than three times revealed significant differences in a dichotic listening shadowing task and symptomatology. The schizophrenic nonresponders as compared to the responders made more errors of omission in the dichotic listening task and showed a symptom pattern of low spontaneous activity and withdrawal. The lack of skin conductance responses in some schizophrenics was interpreted as a protective mechanism.


Panic-fear in asthma. Symptomatology as an index of signal anxiety and personality as an index of ego resources. Clinical observations and studies of asthmatic patients have often concluded that therre is a strong relationship between the degree of the patient's anxiety and the medical intractability of his illness. However, psychotherapeutic interventions designed to alleviate patient anxiety have been noticeably inconsistent in achieving meaningful alleviation of the patient's asthma. The present paper addresses this apparent paradox by positing the existence of two types of anxiety: a) asthma-specific anxiety, as indexed by Panic-Fear symptomatology scores of the Asthma Symptom Checklist; and b) characterological and pervasive anxiety, as indexed by Panic-Fear personality scores of the Minnesota Multiphasic Personality Inventory. In this study, long term medical outcome was found to be influenced by the combination of these types of anxiety. Wehn high asthma-specific anxiety coexisted with high characterological anxiety, medical outcome following intensive long term medical treatment was exceptionally poor. In contrast, when high asthma-specific anxiety coexisted with average levels of characterological anxiety, medical outcome was exceptionally good. These results are discussed relative to the theoretical distinctions between signal anxiety and anxiety concomitant with a lack of basic ego resources.


Norepinephrine turnover in pineal gland and superior cervical ganglia. Changes after gonadotrophin administration to castrated rats. The effects of FSH and LH on norepinephrine (NE) turnover in pineal gland and superior cervical ganglia of castrated female rats were examined by measuring the decline of [3H]NE concentration in the organ 30--120 min after a single i.v. injection of the labeled amine. Both FSH and LH treatment significantly decreased [3H]NE turnover in the pineal gland whereas only FSH accelerated it in the ganglia. As compared to controls, the initial uptake of [3H]NE was lower in the pineal gland of FSH- and LH-treated animals, and higher in the superior cervical ganglia of rats injected with FSH. Neither FSH nor LH modified [3H]NE turnover in heart or adrenal gland, nor changed their endogenous catecholamine content. Pineal monoamine oxidase (MAO) activity type B (assaying by using beta-phenylethylamine as substrate) was decreased by FSH or LH injection. Only FSH modified MAO activity in the ganglia by increasing significantly type A enzyme (assaying by using serotonin as substrate). These results suggest that FSH and LH affect significantly NE metabolism in pineal gland and superior cervical ganglia of spayed rats.


The content and uptake of 5-HT by blood platelets in depressive patients. The obtained results and data of literature indicate the increase (absolute or relative) of the rate of 5-HT uptake by platelets in depressives. These data suggest that the acceleration of 5-HT reuptake in neurones may be one of the reasons of insufficiency of 5-HT-ergic processes in depression of 5-HT type (Asberg, 1976).


Clinical and experimental studies of phenytoin-induced hyperkinesias. Phenytoin administration occasionally leads to the induction of hyperkinetic movement disorders. The pathophysiologic basis of this phenomena is unknown, but thought to be a toxic effect of phenytoin. Study of two cases of this disorder and a review of the literature suggest that antecedant pathologic changes in the basal ganglia are prerequisites for the development of phenytoin-induced hyperkinesias. In an animal model of tardive dyskinesia, phenytoin was found to enhance neuroleptic-induced behavioral supersensitivity but have no effect in control animals. We conclude that phenytoin induced hyperkinesias reflect a specific effect of phenytoin on an abnormal neural substrate and suggest the presence of an otherwise silent pathological alteration of the corpus striatum. The diagnostic value of an episode of phenytoin-induced hyperkinesia is discussed.


EEG sleep patterns in Parkinsonian patients treated with bromocryptine and L-dopa: a comparative study. The nocturnal sleep patterns of six Parkinsonian patients treated with Bromocryptine (2-Br-L-ergocryptine CB-154), a dopamine-like agonist, were compared with those of the same patients under L-DOPA treatment. No significant differences were found between the two groups. It is suggested that Bromocryptine, acting on dopamine receptors in the sleep regulating systems at the reticular level in the midbrain has the same effect on sleep patterns of Parkinsonian patients as L-DOPA.


Ganglioside patterns and cholera toxin--peroxidase labeling of aggregating cells from the chick optic tectum. 2The ganglioside compositions of the chick optic tectum and aggregating tectal cell cultures were examined. Both showed similar trends in changes in ganglioside patterns during development. GD3 and GD1b were the predominant gangliosides early in development, while GD1a and several other multisialogangliosides increased in relative amounts with increasing age in vivo and in vitro. Four gangliosides were present early in development which have not previously been reported. These gangliosides are not present at later developmental times suggesting a possible role for them during the critical early stages of nervous tissue differentiation. Some differences were noted when comparing in vivo versus in vitro ganglioside patterns; these differences may possibly be due to the lack of normal retinotectal connections in the cultures. Cytochemical studies on the localization of the presumed cholera toxin--peroxidase binding site GM1 showed conjugate binding correlates with increasing levels of GM1 in the cultures. In older cultures, the conjugate was uniformly localized on all cells and processes in the aggregates. The conjugate also bound to synaptic membranes and intensely stained the synaptic cleft. This latter observation suggests an enrichment of GM1 in the synaptic cleft region.


Neurotrophic control of 16S acetylcholinesterase at the vertebrate neuromuscular junction. Endplate 16S acetylcholinesterase (16S-AChE) from rat anterior gracilis muscle was assessed, 6 hr to 10 days after denervation, by velocity sedimentation analysis on linear sucrose gradients. The innervating obturator nerve was transected either close (1--2 mm, short stump) or far (35--40 mm, long stump) from the muscle. In both instances, the activity of 16S-AChE gradually decreased and reached approximately the same level (10%--20% of control) by 6 days after denervation. However, enzymatic decay started considerably earlier in short stump (12--24 hr) as compared to long stump (4--5 days preparations, i.e., the time of onset of 16S-AChE loss depended on the length of nerve that remained attached to the muscle. Whether this result extended to other AChE molecular forms (10S, 4S) in muscle endplates could not be determined because, in contrast to 16S-AChE, these forms were also detected in red blood cells (4S) and plasma (10S). Only small amounts of 16S-AChE were found in intact obturator nerves (1/100 of that in gracilis endplate regions). Thus a faster depletion of enzyme from shorter nerve stumps after axotomy could not entirely account for the substantial effect of nerve stump length on 16S-AChE. Since muscle contraction ceases immediately following nerve transection, regardless of nerve stump length, the results can be ascribed to the lack of some neural influence other than nerve-evoked muscle activity. The present findings are consistent with the view that maintenance of 16SAChE at neuromuscular junctions primarily depends on regulatory substances which are conveyed by axonal transport and released from nerve terminals.


Ultrastructure of Aplysia neurons having different degrees of light sensitivity. The relationship between ultrastructure and photosensitivity of pigmented neurons of the abdominal ganglion of Aplysia californica was investigated using electron microscopy and electrophysiological methods. Four identified neurons of similar light microscopic appearance were examined; two are photoresponsive and two are not. Illumination hyperpolarizes both responsive neurons. One of them, R2, requires roughly 100 times greater light intensities than does the other, the ventral photoresponsive neuron (VPN), for similar responses. Two neurons lying adjacent to VPN and similar in appearance to VPN do not have measurable electrophysiological responses to even the highest light intensities. All four neurons contained lipochondria, pigmented organelles associated with the light response. Therefore the presence of these organelles is not the only requirement for light sensitivity in these neurons. Illumination appeared to increase the number of membranous lipochondria in both R2 and the ventral neurons, but only in R2 was this increase significant. Factors such as the concentration of lipochondria near the plasma membrane may affect quantitative aspects of the light response, but in the insensitive cells the lipochondria are apparently uncoupled from other factors required for the light response.


Lobster claw motorneurons respond to contralateral sensory stimuli. Unilateral sensory stimuli in an isolated lobster claw--ganglion preparation elicits soma and axon spikes in homologous motorneurons in the ipsilateral as well as the contralateral hemiganglion. This cross excitation of the motorneurons provides a basis for bilateral reflexes in the claw and is likely mediated through interneuron(s).


Cerebrospinal fluid pulse pressure and intracranial volume-pressure relationships. In six anaesthetised and ventilated dogs the CSF pulse pressure was compared with the volume-pressure response (VPR) during continuous inflation of an extradural balloon. Both pulse pressure and VPR increased linearly with the ventricular fluid pressure (VFP) up to a mean VFP of 60 mmHg. At this pressure a breakpoint occurred above which the CSF pulse pressure showed a steeper linear increase, while the VPR remained constant. It is suggested that the breakpoint is related to failure of autoregulation, and that in non-autoregulating patients the CSF pulse pressure is a better parameter of the clinical state than the VPR.


Conditioning of the H reflex by stimulation of the posterior tibial nerve in Parkinson's disease. The excitability curve of the H reflex conditioned by stimulation of a mixed nerve was studied in eight Parkinsonism patients, before and after L-dopa therapy. There was no significant variation between the two curves. However, there was a reduction of the normal early inhibition of the H reflex conditioned by exteroceptive stimulation. This indicates the presence of alterations in the organisation of the reflex pathways at a spinal level in this disease.


An investigation of the antitumor activity of Micromelum integerrimum (Rutaceae). Extracts of Micromelum integerrimum (Buch.-Ham. ex Coleb.) M. Roem. were fractionated based on in vivo activity in mice in the P-388 lymphocytic leukemia system. Activity in ethanol extracts was concentrated in the chloroform partition fraction, which was further resolved by chromatography on silica gel. The known coumarins, micromelin and scopoletin, were crystallized from the active fractions and demonstrated to have antitumor activities. Micromelin was converted to the corresponding butenolide (deoxymicromelin) which was inactive in the 9KB assay.


Abnormal head movements.

Abnormal head movements. Abnormal head movements have been studied in a variety of diseases using objective recording techniques and the data analysed with respect to the frequency content of the movement. Flopping, nodding, tic, chorea, myoclonic jerks, and most head tremors involve frequencies of approximately 2 and 4 Hz which correspond to the natural fundamental and second harmonic resonances of the head as determined by the mechanical properties of the head/neck system. These findings provide a basis for classification of abnormal head movements as well as an explanation of the characteristics of those arising from hypotonia of the neck muscles. The similarities between tremor frequencies and natural resonances suggest that in the case of the head, tremor arises from disorders of neural mechanisms normally responsible for the fine control of voluntary head movement and for stabilisation of the head during disturbance of posture. Head movements in cases of congenital nystagmus were found to be of two types. Some were of bizarre waveform, in no way assisted vision, and were taken to be of primarily pathological origin and classified as tremors. Others were learned adaptive responses which assisted vision either by interrupting the nystagmus, as in the case of spasmus nutans, or by compensating for the nystagmus with an inverse waveform and were called nodding. A prerequisite for true compensatory nodding is modified vestibulo-ocular reflex.


Assessment of cognitive asymmetries in brain-damaged and normal subjects: validation of a test battery. A test battery designed to assess cognitive functions normally related to the left and right cerebral hemispheres was validated on 30 patients with unilateral (16 right, 14 left) lesions. The tests were preselected to reflect typical functioning of the hemispheres according to general agreement in the literature. A Cognitive Laterality Quotient (CLQ) was calculated from the difference in performance between the "right" and "left" test batteries and, therefore, reflected the relative functioning attributed to the right and left hemispheres. Using the CLQ measurement and a control group of 30 non-neurological patients matched for age and education, 28 out of 30 brain-damaged patients (93%) were categorised correctly according to side of lesion; the other two were considered to have either abnormal lateralisation (one was left handed) or asymmetrical premorbid cognitive profiles. Using only one (paired) test whose two subparts were designed to vary only slightly in task requirements to measure either right or left functioning, 29 out of 30 patients were correctly categorised. It is suggested that the concept of relative assessment of basic cognitive functions is more fruitful than general assessment of intellectual functions for use in diagnosis and rehabilitation of neurological patients or normal subjects with developmental or acquired behavioural cognitive abnormalities.


Antitumor agents 37. The isolation and structural elucidation of isohelenol, a new antileukemic sesquiterpene lactone, and isohelenalin from Helenium microcephalum. The known compound isohelenalin (1) was isolated from Helenium microcephalum and the previously reported structure confirmed on the basis of physicochemical data and chemical transformation. In addition, a new antileukemic sesquiterpene lactone, isohelenol (2), was isolated and its structure elucidated.


Interictal acute psychoses in temporal lobe epilepsy during withdrawal of anticonvulsant therapy. Acute interictal psychotic attacks during withdrawal of medication are described in two patients with temporal lobe epilepsy submitted to depth EEG study with a view to surgical treatment. The patients were on chronic treatment with clonazepam associated in one with phenobarbitone and in the other with phenobarbitone plus carbamazepine. Our observations suggest that the acute withdrawal of clonazepam, the plasma levels of which were monitored may play a part in producing psychotic attacks characterised by dysphoric manifestations, irritability, aggressiveness, anxiety, and hallucinations. These symptoms could be interpreted as a withdrawal syndrome.


First coloured South African with multiple sclerosis confirmed by necropsy. The clinical details and neuropathological findings of a Coloured South African with multiple sclerosis are presented. The interest of this case lies in the fact that this is the first proven case of multiple sclerosis in a Coloured South African.


Lymphocyte function in myasthenia gravis. Mitogen-induced blastoid transformation of peripheral blood lymphocytes from patients with myasthenia gravis was studied using a microplate culture technique and evaluated with 3H-thymidine incorporation. It was found that both phytohaemagglutinin and pokeweed mitogen responses decreased significantly in patients with myasthenia gravis. In myasthenic crisis, indices of stimulation by phytohaemagglutination became very low. The autologous plasma neither inhibited nor facilitated mitogenic responses of lymphocytes. The decreased mitogen responsiveness of lymphocytes suggests that part of the T lymphocyte function is subnormal in myasthenia.


Crossed acoustic response combined with visual and somatosensory evoked responses in the diagnosis of multiple sclerosis. The crossed acoustic response, (CAR), a recently introduced test of brainstem function, has been studied in 66 patients with multiple sclerosis and 53 control subjects, and compared with conventional visual and somatosensory evoked responses (VER, SER). A latency abnormality was found in the CAR in 73% of patients, in the VER in 63%, and in the SER in 37%. Abnormalities have been related to the presence or absence of clinically detectable signs. All three responses detected subclinical lesions by showing abnormality in a proportion of multiple sclerosis patients who had no corresponding abnormal clinical signs (CAR 69%, VER 42%, SER 29%). The best diagnostic combination of responses was VER and CAR. Ninety per cent of patients had at least one of these two responses abnormal.


Cerebrospinal fluid rhinorrhoea from massive osteolysis of the skull. Involvement of the skull by a rare disorder-massive osteolysis-led to CSF rhinorrhoea and meningitis. This is believed to be the first time that such a case has been reported.


Development of mouse dorsal root ganglia: an autoradiographic and quantitative study. Pulse labelling with tritiated thymidine was used to determine the cell birthdays of dorsal root ganglion (DRG) neurons in foetal mice. The peak number of cell birthdays occurred at 11.5 days foetal age in cervical DRGs, and at 12.5 days in lumbar DRGs. The satellite cells were becoming heavily labelled by day 13.5 in lumbar and some hours earlier in cervical regions. A very sharp peak of satellite cell labelling was seen at 13 days in the lumbar region. Evidence for the existence of more than one neuronal cell type is presented. The earliest cells to stop dividing were part of a widely spread distribution which included all the large neurons. The birthdays of the population of small neurons began later and continued for at least 48 h after division of the large cells had ceased.


The postnatal development of large light and small dark neurons in mouse dorsal root ganglia: a statistical analysis of cell numbers and size. A method is described for the analysis of cell types in mouse dorsal root ganglia using the distribution of cell cross-sectional areas measured at the level of the nucleolus in 1.5 micron Epon sections. Using a computer program it was possible to demonstrate the existence of two normally distributed sub-populations of neurons in all the 3rd lumbar segment ganglia (17 in number) measured at various ages from birth to 70 days. The two populations appeared to correspond with large light cells and small dark cells. The large light cell bodies increased in size until about 20 days postnatal, subsequently their size decreased whereas the mean size of the small dark cells reached a plateau by about day 10. The relationship of both nuclear volume and surface area to the surface area of the perikaryon differed between light and dark cells. The number of neurons in L3 remained virtually constant at about 6000 throughout the period examined. Since the proportion of neurons in each population was not shown to change with age there was no evidence that cells could change from one type into the other.


The inner plexiform layer in the retina of the cat: electron microscopic observations. Neural connections of cells ramifying in the inner plexiform layer of the cat retina have been studied by serial section electron microscopy. Flat cone bipolars and invaginating cone bipolars segregate their axon terminals to different sublaminae of the IPL (sublamina a and sublamina b, respectively) where they relate to different subtypes of the same class of ganglion cell (a and b types respectively). Rod bipolar axon terminals end solely in sublamina b and synapse with amacrine cells (AI and AII). AI provides reciprocal synapses to clusters of rod bipolar axon terminals. The AII amacrine provides rod input to a type ganglion cells by means of chemical synapses and to b type ganglion cells through gap junctions with invaginating cone bipolar terminals. Amacrine cells exist which interconnect rod and cone bipolars, but some amacrines appear to be related specifically to neurons branching in particular sublaminae. Both large- and small-bodied ganglion cells have amacrine-dominated input while the medium-bodied ganglion cells with small dendritic trees have cone bipolar-dominated input.


The projection of the lateral geniculate nucleus to area 17 of the rat cerebral cortex. V. Degenerating axon terminals synapsing with Golgi impregnated neurons. The sites of termination of afferents from the lateral geniculate nucleus to layer IV and lower layer III in area 17 of the rat visual cortex have been determined by use of a combined degeneration--Golgi/EM technique. Degeneration of geniculocortical axon terminals was produced by making lesions in the lateral geniculate body. After the animals had been allowed to survive for two days, the ipsilateral visual cortex was removed and impregnated by the Golgi technique. Suitably impregnated neurons and their processes in layer IV and lower layer III were then gold-toned and deimpregnated for examination in the electron microscope. A search was made for synapses between degenerating axon terminals and the gold-labelled postsynaptic neurons. Geniculocortical synapses were found to involve: (1) the spines of basal dendrites as well as those of proximal shafts and collaterals of apical dendrites of layer III pyramidal neurons; (2) the spines of the apical dendritic shafts and collaterals of layer V pyramidal neurons; (3) the perikaryon and dendritic spines of a sparsely-spined stellate cell; and (4) the perikaryon and dendrites of a smooth, bitufted stellate cell. In view of this variety of postsynaptic elements it is suggested that all parts of the perikarya and dendrites of neurons contained in layer IV and lower layer III which are capable of forming asymmetric synapses can be postsynaptic to the thalamic input. Finally, an analysis of the known neuronal interrelations within the rat visual cortex is presented.


Regeneration and myelination in autonomic ganglia transplanted to intact brain surfaces. Fragments of superior cervical ganglia (SCG) from donor rats between newborn and three months of age were transplanted either into the fourth ventricle, onto the dorsal surface of the medulla or in contact with the area postrema of recipient rats aged 6--14 days (allografts) and 3--4 weeks (autografts). Except for the meninges, the entire brain surface and parenchyma was undisturbed. The regenerative capacity of the transplanted ganglia and its interaction with the brain surfaces was followed for post-operative periods between 1 h and six months. Both ependymal and glial cells reacted to the transplant even though there was no mechanical damage to the brain. Ependymal cells developed luminal fronds that projected into the ventricle and the subpial glia displayed a very subtle gliosis in the form of thin multi-laminated processes. Schwann cells from the transplant tended to cover the free surfaces of the brain. The transplants, often incorporated into the stroma of the choroid plexus, received an extensive vascular supply of both fenestrated and non-fenestrated vessels. In contrast to SCG in tissue culture, the perinatal explants quickly degenerated while all those from older donors, at least 3--4 weeks of age, regenerated briskly in the ambient cerebrospinal fluid. Thriving SCG neurons, which diminished in number over time, sprouted numerous neurites as early as one week; growth cones and synaptic contacts between cell processes were still evident at six months. The trasplanted mature SCG fragment underwent a redevelopment after an initial period of degeneration. It seems likely that the survival of the allografted ganglion cells depends on their acquisition of a target site in their new environment. By four to six months many axons became enclosed by myelin produced by SCG Schwann cells that normally do not form myelin in situ. Other Schwann cells appeared reactive in that they had a great increase in cytoplasmic filaments and formed gap junctions, two characteristics of C.N.S. astrocytes. It is possible that the proximity to the C.N.S. changes the character of certain Schwann cells or, alternatively, resulted in the migration of glial cells out of the brain. If the glial cells have migrated into the transplant, they may support alien neural tissue. This system in which the transplantation site is easily accessible with a minimum of trauma could lend itself to the study of some underlying mechanisms of the growth and regulation of both central and autonomic neurons and their supporting cells.


Structure of rat aortic baroreceptors and their relationship to connective tissue. The ultrastructure of fibres and sensory terminals of the aortic nerve innervating the aorta between the left common carotid and left subclavian arteries was investigated in the rat. This is the region from which most baroreceptor responses are recorded electrophysiologically. The fibres of the aortic nerve enter the adventitia and separate into bundles generally containing one myelinated fibre and four or five unmyelinated fibres of various sizes. The bundles pursue a roughly helical course through the adventitia; when they are close to the aortic media, the myelinated fibre loses its myelin sheath. A complex sensory terminal region is formed, as both the unmyelinated and 'premyelinated' axons become irregularly varicose. The concentration of mitochondria becomes very dense and cytoplasmic deposits of glycogen are observed. Both unmyelinated and premyelinated axons branch, and the unmyelinated axons wind irregularly around the premyelinated axon. The latter may have several loops and small holes. The terminal regions of both types of axon contain clusters of clear 40 nm vesicles. Part of the surface of each terminal region is ensheathed by Schwann cells, but the rest of the axolemma is directly exposed to extracellular connective tissue. There are often several layers of basal lamina around the sensory terminals and parts of the axolemma and Schwann cell membranes are attached to it by fine fibrillar material. The basal laminae are also attached to fibroblasts, fibroblast-like perineurial cells and elastic laminae, and the whole cellular and extracellular system appears to be tightly bound together. No differences between baroreceptors of spontaneously hypertensive and normal rats were found.


Ultrastructure of the synapses of sensory neurons that mediate the gill-withdrawal reflex in Aplysia. We have identified the processes of mechanoreceptor sensory neurons by intracellular injection of horseradish peroxidase in order to study the structure of synapses which exhibit profound, behaviourally-relevant plasticity. These synapses are located at small, varicose expansions along or at the end of the fine, microtubule-containing neurites, and they are crowded with vesicles some of which are associated with the varicosity membrane at regions of membrane specialization morphologically equivalent to active zones described in other species. These active zones occur between pre- and postsynaptic processes at two varieties of apposition: a conventional flat one, and a more elaborate indented one. At indented appositions, the presynaptic varicosity is invaginated by a thin (less than 0.25 micrometer diameter) spine of variable length. The active zones of indented synapses have approximately twice the vesicle frequency of flat synapses, suggesting that indented synapses are more effective. Sensory neuron terminals are relatively uniform in their structure, having similar concentrations of vesicles and numbers of active zones, and the majority of the processes postsynaptic to them are less than 0.5 micrometer in diameter. These regularities, and the presence of two strikingly-different types of synaptic apposition, flat and indented, should facilitate structural comparisons of neurons from naive and behaviourally-modified animals. The possible dynamic interconversion of indented and flat appositions at the synaptic terminals of sensory neurons and its behavioural relevance are discussed.


Serotoninergic synapses on ependymal and hypendymal cells of the rat subcommissural organ. The nervous input to the subcommissural organ (SCO) of the rat has been investigated with Falck-Hillarp fluorescence histochemistry and electron microscopical techniques. Previous fluorescence histochemical observations of a dense plexus of serotoninergic nerve fibres in relation to the basal SCO were confirmed. Electron microscopically, unmyelinated fine varicose axons ranging in size from 0.1--0.6 micrometer were observed to penetrate into the SCO hypendyma. Boutons and presynaptic varicosities filled with a diversity of round and elongated clear vesicles, and occasional large dense cored vesicles establish asymmetric (Gray's type I) synaptic contacts with the basal processes and somata of the SCO ependymal and hypendymal cells. A typical varicosity in synaptic contact with an SCO cell contains a population of approximately 85% clear, elongated vesicles 45 X 60 nm in diameter, 15% clear, round vesicles 50 nm in diameter, and 1--2% large dense cored vesicles with a vesicle diameter of about 85 nm and a dense core diameter of 50--55 nm. The mean length of the postsynaptic membrane specialization was found to be 0.5 micrometer. Experiments with specific neurotoxic drugs revealed that the nerve terminals in synaptic contact with the SCO cells are identical to the fibres of the serotoninergic plexus identified fluorescence histochemically. Thus, an intraventricular injection of either 5,6-dihydroxytryptamine or 5,7-dihydroxytryptamine induced typical degenerative changes in most of the boutons in synaptic contact with the SCO cells, and also a disappearance of the yellow fluorescent nerve plexus. It is concluded that the SCO of the rat receives a dense plexus of serotonin-containing nerve fibres which form typical synaptic contacts with the specialized ependymal cells of the SCO and that these fibres may constitute the only direct nervous input to the organ. The degeneration of the serotoninergic synapses elicited a long-lasting, pronounced increase in the secretory activity of the SCO. Despite long survival times after the treatment with neurotoxic drugs, we found no evidence of regenerative restitution of the serotoninergic innervation nor normalization of the secretory activity of the SCO. The observed inverse relationship between secretory activity and serotoninergic innervation is in line with previous observations which indicate that the 5-hydroxytryptamine input to the SCO ependymal and hypendymal cells exerts a powerful inhibition on their protein synthetic machinary.


Neurotoxic destruction of the serotoninergic innervation of the rat subcommissural organ is followed by reinnervation through collateral sprouting of non-monoaminergic neurons. Specific neurotoxic destruction of the serotoninergic innervation of the subcommissural organ of the rat is followed by an efficient reinnervation by collateral sprouting of non-monoaminergic axons, which normally do not innervate the SCO cells. Morphologically, the reinnervating fibres totally replace the serotoninergic synapses lost by the lesion, but, functionally, they fail to substitute for the potent inhibitory control of secretory activity normally exerted by the serotoninergic innervation. It is possible that the observed reinnervation by foreign synapses explains why the regrowing serotoninergic neurons fail to re-establish their connections with the SCO.


The tight epithelium of the Mongolian gerbil subcommissural organ as revealed by freeze-fracturing. Examination of neonatal Mongolian gerbil subcommissural organ (SCO) by freeze-fracture electron microscopy reveals that the epithelial layer (the ependyma) possesses distinctive and very complex tight junctions which connect the apical portions of adjacent ependymal cells. The tight junction network of the Mongolian gerbil is more extensive neonatally than in the adult, and is in general, more conspicuous than in other animal species. The consistent finding of such very complex tight junctions at the interface between the C.S.F. and the SCO suggests that the SCO ependyma, at least in neonatal Monglian gerbils, might function as a tight epithelium with an ability to maintain steep solute gradients and high potential differences. The sealing-off of the SCO towards the C.S.F. (the C.S.F.--SCO barrier) and the effective blood-brain barrier in the organ (the blood-SCO barrier) might well be of functional significance, perhaps in relation to mechanisms involved in the control and/or detection of the stability of the internal environment of the brain.


Fine structure of initial outgrowth of processes induced in a pheochromocytoma cell line (PC12) by nerve growth factor. Cells of the PC12 line (which is derived froma rat pheochromocytoma) develop neuron-like processes upon exposure to nerve growth factor (NGF), and thus provide an opportunity to study this phenomenon de novo. We have used the transmission electron microscope to analyse the early stages of process outgrowth (1, 2, 3 and 7 days) to determine what organelles are involved and in what sequence they appear during development. Despite the non-synchronous response to NGF, we can derive three main stages in early process formation. (1) NGF-treated cells develop conical extensions similar to, but larger and more numerous than those of controls. Extensions terminate in bulbous expansions that contain large number of chromaffin-like granules and bear microspikes filled with microfilaments. (2) The extensions of NGF-treated cells then acquire membranous organelles indicative of transmitter packaging and/or recycling of cytoplasmic membranes, for example, tubular reticulum, clear and dense-cored vesicles, multivesicular bodies, and lysosomes. (3) As processes elongate, they develop a shaft that contains an array of microtubules and fine tubular reticulum dispersed in a filamentous matrix, and varicosities that exhibit the same organelles seen in stage 2. The discussion stresses the similarities in the outgrowth of processes in PC12 cells and neurons, and speculates that NGF causes a change in organization and/or quantity or organelles that already exist in non-treated control cells.


Immunohistochemical localization of neurofilament antigen in rat cerebellum. The distribution of neurofilaments in the rat cerebellar cortex was studied by immunoperoxidase histochemistry using an antiserum raised against neurofilaments isolated from brain (anti-NF). In light microscope preparations, this antiserum selectively stained known neurofilament-containing structures. Staining was most intense in myelinated axons of the white matter and in the terminal branches of basket cell axons. No staining was apparent in either neuronal or glial cell bodies or in glial cell processes. These findings were confirmed in electron microscopic preparations of the same material. Neurofilaments stained by the antiserum were abundant in basket cell axons and also occurred in small bundles in mossy fibre terminals. Adjacent microtubules were not stained by the antiserum. There was no evidence of stained cytoplasmic filaments in glial cell processes. Thus it appears that neurofilaments contain unique antigens which do not occur in either microtubules or in glial cytoplasmic filaments. The antiserum did not induce staining of synaptic junctional structures, a result which contradicts previous suggestions that neurofilaments are structural components of synaptic densities.


Cross-correlation analysis of connectivities among cat lumbosacral dorsal horn cells. 1. Eighty-three cell pairs were recorded in dorsal horns of 19 cats. Cross-correlograms were flat in 65 cell pairs; of the remaining 18 pairs, cross-correlograms had broad peaks or troughs suggestive of nonmonosynaptic causal relations in their discharges; one cell pair had a biphasic deviation from base line. 2. No cross-correlograms indicative of monosynaptic interactions were observed. 3. All cell pairs with nonflat cross-correlograms had overlapping receptive fields. 4. It is concluded that monosynaptically connected cell pairs are rare at the interelectrode distances used in this experiment because such cell pairs must lie close together on the somatotopic map or because our recording methods bias against monosynaptically linked cell pairs.


Neuronal mechanisms of human locomotion. 1. The surface electromyogram (EMG) of human leg muscles was recorded during running at different speeds; The EMG was compared with the simultaneously recorded vertical force exerted by the foot and with the angle of the ankle joint. 2. During running, the electrical activity of the gastrocnemius muscle increased sharply 35--45 ms after ground contact and reached its maximum at the end of muscle stretch; This activity was superimposed on a slowly increasing level of activation, which began 120--180 ms before ground contact. At the end of the stance phase, gastrocnemius became inactive and, simultaneously, there was a sudden increase in tibialis anterior activity. The assumption that the steep increase in the gastrocnemius EMG reflects the spinal stretch reflex of alpha-motoneurons is supported by the following findings. 3. The peak level of gastrocnemius EMG in the stance phase of fast running was 2--3 times higher than the activity during maximum voluntary contraction. 4. With stimulation of the tibial nerve at different rates, the maximum isometric force was about 30--40% higher than the maximum voluntary isometric contraction. 5. The increase in EMG at 35--45 ms after ground contact was markedly diminished during running, after partial blockage of Ia afferents by ischemia, at a time when the strength of voluntary contraction was shown to be uninfluenced by the ischemia. 6. The gastrocnemius activity during running was simulated by electrical stimulation of the tibial nerve. The rate of stimulation was varied so as to approximate to the EMG profile during running. This indicated that a spinal stretch reflex could become mechanically effective within the shortest stance phase measured in a fast sprint (ca. 120 ms).


Neural correlates of nystagmus in abducens nerve. 1. The firing rates of action potentials of abducens nerve single fibers were recorded in the cat's orbit during a variety of vestibular and optokinetic stimulations. 2. Comparison was made of the neural firing rates associated with agonist and antagonist responses during slow and fast components of vestibular and optokinetic nystagmus. It was found that the relationship between the motoneuron firing rates and the eye motion was independent of the reflex with which they were associated--vestibular or optokinetic, or the type of response--agonist or antagonist. No neurons were observed that responded only during the fast or only during the slow nystagmus phase. Motoneuron firing rates were proportional to both velocity and position of the eye in a ratio of 1 (spikes/s)/(deg/s) to 7.2 (spikes/s)/deg. The behavior of the motoneurons was compatible with the hypothesis that thier firing rates are sufficient to overcome both elastic and viscous forces by which the muscles and ligaments hold the eye in the orbit. 3. For low-frequency head rotations, eye displacement and neural responses showed a small phase angle difference. At higher frequencies, however, while the eyes maintained a fixed relationship to the head rotation, the neural responses showed an increasing phase lead. One component of this phase lead compensated for the phase lag introduced by the orbital mechanics. The other was modeled as a constant delay of approximately 70 ms, which may be accounted for by neuromuscular transmission and transduction.


Concept of neuron types in gustation in the rat. 1. In taste neurophysiology, from Pfaffmann's (49, 50) pioneering work until the present, the possibility of types of neurons corresponding in some sense with the "primary" taste qualities of Henning (33) has been entertained: recently types of gustatory neurons in peripheral nerves have been established according to which of the four classical stimuli is the "best stimulus." However, considerable variation occurs in the response profiles within neurons classified as belonging to the same type. The purpose of this research is to determine, using mathematical techniques where appropriate, if the within-type variation is spurious or, instead, indicates the absence of a typology of taste neurons. The data used were counts of the spike discharges of 50 individual taste neurons in the nucleus of the solitary tract of the rat, evoked by 32 diverse chemical stimuli. 2. Using as input the matrix of Pearson r correlation coefficients calculated for the responses of all pairings of neurons to all stimuli, multidimensional scaling analysis revealed a two-dimensional space in which no clear groupings of neurons occurred. 3. In a hierarchical cluster analysis of the neuron response profile similarities, no evidence of grouping was found, suggesting a more-or-less continuous variation among neurons. 4. When the organization of the 32 stimuli utilized was studied by the same techniques, no clear evidence for stimulus types was found, although the possibility of two stimulus types--"sweet" and "nonsweet"--was raised. 5. Construction of a joint neuron-stimulus space supported a spatial model of taste neuron-stimulus interaction, while analysis of the number and pattern of high correlations among neurons--even after allowance for attenuation due to measurement error--failed to support the notion of types of taste neurons with identical response profiles. 6. Aspects of the logical role of types of neurons in gustatory coding were discussed, and the results and methods of the present investigation were related to classification schemes for neurons in general. Suggestions for a formal taxonomy of neurons were given. 7. It should be emphasized that the present study and conclusions are of second-order, CNS neurons, whereas the studies advocating the presence of neurons types were of peripheral neurons. Taken together, the implication to be drawn from these studies is that if neural types do exist in peripheral taste nerves, the typology is lost at the first synapse and is thus unavailable to the CNS for coding purposes, at least in the rat.


Guidance of regrowing sensory axons after cutaneous nerve lesions in the cat. 1. Individual type I sensory neurons in cutaneous nerves typically innervate two to four type I cutaneous mechanoreceptors (Haarscheiben). The extent to which these neurons replicate the original innervation patterns of the type I receptors after peripheral nerve regeneration and the means by which these neurons are guided back to their old receptor sites during regeneration were studied in cats using neurophysiological techniques. 2. By recording activity of type I neurons in small cutaneous nerves and isolated dorsal rootlets, it was possible to map the distribution of these neurons in the skin. Maps made before nerve lesion were compared to maps made after recovery from nerve crush and transection. 3. Fibers regenerating after nerve crush return to their old receptor sites, probably by following their old Schwann tubes in the distal stump of the nerve, and replicate the original receptor innervation pattern. Essentially all the type I fibers successfully regenerate in this case. 4. In contrast, after nerve transection the regenerating fibers do not restore the original innervation pattern, although they do preferentially return to other old type I receptor sites. About 60% of the type I fibers reinnervate the skin after transection. 5. These observations provide a basis for the difference in functional recovery seen after crush and transection lesions of peripheral nerves.


Spatial and temporal features of afferent inhibition of thalamocortical relay cells. 1. The temporal and spatial features of the afferent inhibition of thalamocortical relay (TCR) cells in the ventrobasal complex of the thalamus have been examined using paired conditioning and test air jets. 2. The response of TCR units to air jets may be divided into three parts: a) an early response of 1--6 impulses, which begins after a latency of 8--10 ms and lasts for 10--25 ms; b) a period of 70--80 ms following the early response, during which the spontaneous activity is inhibited; and c) a period of late activity, which follows the inhibitory period. 3. The inhibition of TCR units generated by an air jet lasts about 80 ms. The time course of inhibition is the same in hair units and in units activated by Pacinian corpuscles. Evidence is presented that suggests that inhibition decays at the same rate throughout the inhibitory receptive field. 4. The spatial features of inhibition demonstrated in this study are: a) that the excitatory and inhibitory receptive-field centers coincide; b) that these fields have different shape; and c) that there is a significant area in which the inhibitory receptive field surrounds the excitatory receptive field.


Statoacoustic properties of utricular afferents. 1. We classified the utricular afferents on the basis of their spontaneous acitivity and responses to tilts and vibrations. 2. Type I afferents fire spontaneously in a regular pattern; their responses to tilts consist of a phasic-tonic change in firing rate. They may respond to vibrations by increasing or decreasing their rate and show no adaptation. 3. The spontaneous activity and the responses to tilts of type II are similar to those observed in type I afferents. The differences become apparent when the preparation is subjected to a vibrational stimulus, since type II neurons increase their firing rate regardless of the stimulus frequency and show adaptation. 4. Type III neurons have no spontaneous activity. They respond to tilts by firing during the transition from one position to the other. They respond to a vibrational stimulus with maintained firing and show no adaptation. 5. We studied the dynamic responses of each type of neuron. We used sensitivity curves for the study of type III afferents and proposed a statistical method to define gain curves for the study of the other types. 6. The gain curves generated by type I neurons reach their maximum at frequencies of stimulation close to the spontaneous rate of firing. 7. In the gain curves of type II afferents the maximum corresponds to frequencies higher than their spontaneous activity. 8. Sensitivity curves and gain curves give similar results for type III fibers. The sensitivity curves of these afferents were classified into four subtypes. 9. We studied the responses of the three types of afferents to bursts of sinusoidal vibrations. 10. We concluded that the properties of types I and II fibers are fit to carry information about movements and position of the head, but also transmit acoustical information. Type III fibers are more adapted to provide information about acoustical stimuli, but can also convey information about head movements.


Hepatic-portal glucose and insulin levels: relationship to glucose-induced satiety and hunger. The hepatic-portal infusions of both smaller and larger glucose loads, which had no significant effect on food intake in chronic feeding experiments, produced only a brief dose-dependent peak-like elevation in glucose and insulin levels. In contrast, metabolic changes produced by duodenal infusions were greater and considerably longer in duration. The maximal level and duration of glucose and insulin increase further differentiated two feeding phenomena induced by duodenal infusions: 1) the metabolic satiety of smaller glucose loads associated with blunted and delayed glucose and insulin increases within normal physiological range; 2) paradoxical antiregulatory stimulation of food intake induced by larger glucose loads associated with a gradually intensified, exaggerated and prolonged insulin increase. This increase was significantly larger than corresponding preinfusion value even at the end of 3 hour test period. Present results thus demonstrate that various glucose-induced feeding phenomena are associated and apparently dependent for their expression on distinct, prolonged changes in glycemia and insulin levels.


The absorbability by rats of various triglycerides of stearic and oleic acid and the effect of dietary calcium and magnesium. Rats were fed diets in which the triglycerides contained oleate and stearate as the sole fatty acids. These fatty acids were esterified to specific positions on the glycerol molecule. The triglycerides were 1-stearoyl diolein (SOO), 2-stearoyl diolein (OSO), 2-oleoyl distearin (SOS), 1-oleoyl distearin (OSS), and triolein (OOO). The absorbability of the fatty acid component was measured by the fat balance technique. Two diets, one sufficient and the other deficient in calcium and magnesium, were used. The oleic acid of all of the triglycerides was absorbed almost completely. The following values for the absorbability of the stearate component in the presence and in the absence of the divalent cations were obtained: OSO 98 and 99; SOO 55 OAND 96; SOS 37 and 70; OSS 59 and 60. These patterns of absorbability are discussed in relation to the pathway of triglyceride digestion. If the stearic acid is esterified at the 2-position of the triglyceride, the resulting 2-monostearin is well absorbed. If it is esterified at the 1- or 3-position, it is released as free stearic acid, and in the presence of calcium and magnesium it is poorly absorbed.


Influence of dietary zinc on lead toxicity during gestation and lactation in the female rat. Confirmed pregnant female albino rats received a purified diet containing either 12 or 120 ppm zinc with or without lead (0 to 500 ppm) from day 0 gestation through day 16 of lactation. An 11% reduction in dam weight gain during gestation and a 12% reduction in average pup weight in rats fed the low zinc plus lead diet was markedly different compared to controls not receiving lead and lead supplemented rats receiving supplemental zinc. Milk lead concentration was reduced in rats receiving additional dietary zinc and lead which was reflected in a 15% reduction in pup tibia lead concentration, a 20% reduction in inhibition of pup liver delta-aminolevulinic acid dehydratase activity and a 31% reduction in pup blood porphyrin concentration. Some of the beneficial effects of supplemental zinc on lead toxicity of pups may be explained by a reduction of both liver lead (40%) and blood lead (32%) concentration of dams. However an interaction between lead and zinc at the placental and mammary barriers must also be considered. The results of this study suggest that nutritional status of zinc may be an important factor in modifying lead burden of infants borne by women occupationally and environmentally exposed to lead.


Estimation of nutrient requirements from growth data. Two least squares methods of estimating nutrient requirements from growth data were compared. One method involved fitting a broken line by the method of least squares. The requirement was taken as the abscissa of the breakpoint in the curve. The other method involved fitting an appropriate exponential function to the growth data and estimating the requirement as the abscissa of the point on the fitted curve whose ordinate was 95% of the upper asymptote. For the nine sets of data studied, the broken line provided adequate fits for only six. The nonlinear models provided adequate fits for all the data studied. When both the broken line and the chosen nonlinear model provided adequate fits, the estimated requirements were nearly the same. However, the consistently good fits obtained with the nonlinear models suggest that this approach may generally be more useful.


Accumulation and depletion of zinc in chick tissue metallothioneins. Two experiments were conducted with Hubbard broiler chicks on the metabolism of high levels of dietary zinc. In the first experiment, chicks were fed a basal diet or the basal diet plus 500, 1,000, 2,000 or 4,000 ppm zinc, and in the second experiment chicks were fed the basal diet or the basal diet plus 1,000, 2,000, 4,000, 8,000 or 16,000 ppm zinc, using zinc acetate. Zinc was found to be associated with 4 peaks when the cytosols from tissues (liver, kidney, pancreas and intestinal mucosa) were eluted through columns of G-75 Sephadex. Although the zinc content of all four peaks increased with increased zinc content of the diet, the greatest elevation occurred in metallothionein (MT). This protein was purified from all four tissues (liver, kidney, pancreas and intestinal mucosa) and the amino acid analysis revealed the presence of about 30% cysteine. Zinc accumulated to the greatest extent (microgram in MT/g tissue) in MT of the pancreas. The zinc in MT disappeared very rapidly when chicks which had been fed a diet containing high levels of zinc were fed a low zinc diet, indicating the extreme lability of this metal in this protein. The results suggest the involvement of MT in zinc homeostasis.


Effects of dietary 3-methylthiopropionate on metabolism, growth and hematopoiesis in the rat. Rats were fed a basal diet supplemented with (2.57%) 3-methylthiopropionate (MTP) for 2 weeks. A marked depression in growth and food intake similar to that found in rats fed an equimolar level of methionine was observed. While supplemental glycine or serine alleviated the toxicity due to dietary methionine, similar levels added to the diets of rats fed MTP were without effect. The spleens of rats fed diets containing 2.57% MTP were grossly enlarged and darkened in comparison to spleens from control rats and histological examination of these spleens by light and transmission electron microscopy (TEM) revealed sequestration of large numbers of erythrocytes in the splenic sinusoids and red pulp similar to that seen in rats fed high levels of methionine. Marrow changes included increased numbers of erythroblastic islets and subtantial electron dense hemosiderin deposits in islet reticulum cells. Examination of peripheral blood erythrocytes by scanning electron microscopy revealed extensive variation in the size of the erythrocytes and the presence of large numbers of misshapen red cells in rats fed the diets containing MT. When viewed by TEM many erythrocytes had obvious membrane defects and remnants of cytoplasmic organellae. Many erythrocytes with reticulocyte morphology were present in the peripheral blood. This condition is characteristic of maturation arrest at the reticulocyte stage of development. The similarity of depression in growth and food intake and the identical abnormalities found in the spleens of rats fed high levels of MTP and methionine suggest that the transamination pathway of methionine catabolism may be important with respect to the toxicity of methionine. The ultrastructural changes noted in MTP-fed rats suggest a serious dysfunction of red cell hematopoiesis. The large numbers of defective and/or immature erythrocytes released from the marrow into the peripheral circulation, only to be later sequestered and destroyed in the spleen, is a reflection of a serious derangement.


The relationship of plasma arginine and kidney arginase activity to arginine degradation in chickens. Experiments were carried out to study urea excretion during arginine or ornithine infusion into wing veins of hens previously fed diets that induced different arginase levels in their kidneys. Urea excretion was found to increase as plasma arginine increased. Hens with high levels of arginase activity in their kidneys had a greater increase in urea excretion than hens with low kidney arginase activity. Arginine degradation was also dependent on both the kidney arginase activity and on the plasma level of arginine. Ornithine infusion did not inhibit urea excretion even when high levels of plasma ornithine were reached. Even though ornithine was an in vitro inhibitor of arginase, no evidence was obtained of in vivo inhibition.


On the physiological basis of vitamin A-stimulated growth. The influence of vitamin A depletion on tissue composition was studied in rats that were marginally vitamin A deficient, i.e. at their weight plateau stage. The total number of cells (DNA) was decreased in most organs as a result of vitamin A depletion. In thymus, spleen and the sublingual glands there was also a dramatic reduction in the number of cells per gram of tissue and in thymus and sublingual glands there was an increase in the protein to DNA ratio as a result of absence of dietary.vitamin A. We present the hypothesis that vitamin A stimulates growth by a direct role in cell replication in addition to or instead of stimulating the differentiation of epithelial and bone cells.


Fasting and diet affect the tolerance of young chickens exposed to acute heat stress. The interaction of nutritional status on the tolerance of young chicks to acute heat stress (HS) was investigated. The results of experiment 1 demonstrated that fasting for 24, 48 or 72 hours progressively increased survival time (ST) of chicks exposed to HS. When compared to that of non-fasted (NF) chicks ST more than doubled for 48 and 72 hour fasted chicks. During HS plasma glucose increased in fasted chicks but decreased in NF chicks. Conversely, plasma free fatty acids decreased in fasted chicks and increased in NF chicks. Plasma phosphate of all chicks fell markedly as a result of HS. Chicks of experiment 2 were fed a "carbohydrate-free" diet (CF). Survival time of NF chicks fed CF was significantly greater than that of NF chicks fed a diet containing glucose. The apparent metabolic responses of NF chicks fed CF corresponded to those of fasted chicks of experiment 1. It was suggested that the metabolic substrate utilized by chicks during HS influences ST. The effect of CF diet on ST was also observed in chicks which were fasted for 48 hours prior to HS. Plasma phosphate of chicks exhibiting fasting metabolic responses was not significantly different at heat induced prostration despite marked differences in ST. The results of experiment 3 demonstrated that ST of fasted (24 hour) chicks previously fed a low phosphorus diet was markedly lower than compared to that of fasted chicks fed adequate dietary phosphorus. The effect of diet was found to be dependent on metabolic status since level of dietary phosphorus fed had no effect on ST of NF chicks. The results of the present study indicated that metabolic and nutritional status affect the tolerance of young chicks acutely exposed to high ambient temperature.


Carcass deposition of dietary long-chain odd carbon fatty acids by rats and their effect on plasma glucose and ketone bodies during starvation. Weanling rats were fed diets containing various levels (0 to 40% of total dietary acids) of long chain, odd-carbon fatty acids (OCFA, 15:0 + 17:0) for 5 weeks. The OCFA did not significantly alter growth or feed efficiency and the OCFA were deposited in the carcass fat in proportion to their concentration in the diet fat. After the 5-week ingestion period, the rats were starved for 48 hours and the effect of carcass OCFA content on weight loss, fat loss, urinary total nitrogen, plasma glucose concentration and plasma ketone body concentrations was determined as a function of starvation time. The results demonstrate that OCFA catabolism during starvation results in a dose related increase in plasma glucose and dose related decrease in plasma ketone bodies without significantly altering the total weight loss, carcass fat loss, or urinary total nitrogen. Finally, the carcass percentage of OCFA did not change during starvation showing that these acids are as readily lost from the carcass during starvation as even chain fatty acids.


The effect of vitamin A deficiency on the in vitro cellular immune response of rats. The effect of vitamin A deficiency on the response of splenic lymphocytes to mitogenic stimulation was determined in an experimental rat model. Male Lewis rats were divided into three groups. The ad libitum group (AL) was fed unlimited amounts of a vitamin A-supplemented diet. The vitamin A-deficient group (DEF) received a commercial vitamin A-free diet. The pair-fed group (PF) received a vitamin A-containing diet equivalent in amount to that consumed by the DEP group. During the early stages of vitamin A deficiency (determined by cessation of weight gain), the rats were killed and the isolated splenic lymphocytes subjected to mitogenic stimulation. Lymphocytes from DEF rats had one-third the transformation response to the mitogens Concanavalin A, Phytohemagglutinin and E. coli Lipopolysaccharide S of the AL and PF groups. When the DEF rats were supplemented with vitamin A, the transformation response returned to control values within 3 days. In addition to the alterations in the immune response, the DEF rats showed a marked leukopenia, a decrease in the number of circulating lymphocytes and an increase in the number of circulating neutrophils.


Risks to the offspring from parental occupational exposures. Risks to the offspring of workers with occupational chemical exosures may derive from mutagenic, teratogenic or carcinogenic effects of industrial agents to which the parents are exposed. Evidence for impaired pregnancies and hazards to the offspring of working populations with chemical exposures is, however, very limited. Perhaps the best documented example is increased spontaneous abortion rates in female operating room personnel who have first trimester exposure to waste anesthetic gases. Evidence is reviewed for hazards to the offspring resulting from parental occupational exposure to vinyl chloride, benzene, chloroprene, radiation and petroleum-derived hydrocarbons. It is essential in investigating the role of occupational factors that other environmental and behavioral factors with major effects on pregnancy outcome be accounted for. These include smoking, alcohol, and drug exposures. An approach to surveillance for chromosomal abnormalities in offspring of occupationally exposed parents is outlined.


Skin carcinogenicity of synthetic and natural petroleums. In a series of three separate experiments mice were exposed to various concentrations of fossil liquids obtained from coal, oil shale or natural petroleum. All materials were capable of inducing squamous cell carcinoma, but potency differed substantially. Skin carcinogenicity was markedly greater for both coal or oil shale liquids than for natural petroleums. None of the syncrudes approached the skin carcinogenicity of a pure reference carcinogen, benzo(a)pyrene (BP). It is unlikely that determination of the concentration of an active compound in material applied to the test animal will allow meaningful comparison among the diverse agents of interest to the synthetic fuels industry. To better establish the relationship between actual tissue dose and surface concentration the authors are investigating various in vitro and biochemical measures of hydrocarbon-skin interaction to determine which, if any, could serve as a more definitive measure of surface dose. Results, using BP as a marker carcinogenic hydocarbon, suggest that carcinogenic crudes inhibit both BP metabolism in skin organ culture and the interaction of BP adducts with epidermal DNA, in vivo.