The clinical pharmacology of meperidine--comparison of routes of administration. The serum time-concentration curves and pharmacokinetic parameters of meperidine after the intravenous, intramuscular, and oral administration of 26 mg/m2 doses were determined and compared. After 30 minutes, there was no statistically significant difference between the serum time-concentration curve following intravenous administration and intramuscular administration; and after 2 hours, all three serum time-concentration curves were the same. Comparison of the mean cumulative excretion of meperidine and normeperidine following all three routes of administration demonstrated that the maxmimun rates of both  meperidine and normeperidine reflect the route of administration, with a marked delay following oral administration reflecting a delay in absorption. Further, the excretion of normeperidene following oral administration, is greater despite adequate absorption than the other two routes of administration, indicating that delayed absorption increases the metabolic phase. Subjective symptomatology and the serum time-concentration curves were compared and their relationship wxamined. Comparison of the serum time-concentration curves to pervious analgesic and toxicity trials was made, and minimum serum levels for induction of analgesia and  production of side effects are discussed. From the data it appears that in the case of acute pain, the intramuscular route of administration is as beneficial as the intravenous route, that oral dosing is less efficacious due to lower peak serum concentrations, and that all doses, regardless of the route administration, should be administered clinically on a mg/m2 basis rather than as a predetermined dose as currently used. Oral administration may have some benefit in the treatment of chronic pain, but the possibility of cumulative toxicity due to normeperidine should be considered.


Sustained-release lithium carbonate in double-blind study: serum lithium levels, side effects, and placebo response. The utility and side effects of sustained-release lithium carbonate (Priadel) in a once-per-day dose regimen was investigated with 66 male delinquents, ages 17-24 years, in a double-blind study comparing the antiaggressive effect of lithium carbonate with placebo. Serum lithium levels and symptoms were determined weekly for up to eight drug-free and 12 on-medication weeks. Average daily doses of 1500-1700 mg Priadel gave 24-hour serum lithium levels in the range 0.7-0.9 mEq/liter. Principal side effects were polyuria and shakiness, with other important side effects bring hand tremor, dryness of mouth, nausea, and weakness. No lithium toxicity was observed, and diarrhea was reported infrequently. Placebo response data are presented.


Autoradiographic studies of the projections of the midbrain reticular formation: ascending projections of nucleus cuneiformis. The ascending projections of the cuneiform nucleus in the cat were traced by autoradiography in the transverse and sagittal planes following stereotaxically placed injections of (3)H-leucine. The ascending fibers are almost exclusively ipsilateral and enter the diencephalon as a wide radiation. At the mesodiencephalic junction fibers enter the nucleus of the posterior commissure and pretectal nuclei, and others cross in the posterior commissure to distribute to these structures on the contralateral side. More ventrally directed fibers distribute to the fields of Forel and then spread into the posterior hypothalamus and zona incerta. At the caudal level of the ventral thalamic group, the ascending fibers diverge and follow two separate courses. One division of fibers continues forward beneath the ventral thalamic group and distributes to the zpna incerta and dorsal hypothalamic area. It rapidly diminishes in size as it attains more rostral levels where it is found in the bed nuclei of the stria terminalis and the anterior commissure. Other fibers of this division spread laterally to innervate the ventral lateral geniculate nucleus, the lateral hypothalamus, and preoptic area, and still others follow the entire confirmation of the thalamic reticular nucleus. The second division of fiber ascends through midline and intralaminar nuclei, completely encircling the mediodorsal nucleus, which is uninnervated except for a small ventral region. The distribution of this division is heaviest to the paraventricular, parafascicular, and central dorsal nuclei. Neither division is conspicuous rostral to the anterior commissure. No projections to neostriatum or specific thalamic nuclei were evident.


Failure of vitamin C to affect the pharmacokinetic profile of antipyrine in man. A possible interaction of vitamin C with the metabolism of drugs which are oxidized (e.g., antipyrine and diphenylhydantion) is indicated from data in animals. The effect of megadoses of vitamin C (300 mg/day to 4800 mg/day) was therefore tested in human volunteers receiving antiprine (single oral dose) or diphenlhydantoin (chronic therapy). The half-life, volume of distribution, and plasma clearance of antipyrine were not affected by vitamin C treatment. Steady-state plasma levels of diphenylhydantoin in two epileptic patients were similar before and during vitamin C (1.2 Gm/day for 18 days) treatment. It is concluded that vitamin C, in commonly used megadoses, has no effect on the metabolism of antipyrine or diphenylhydantoin in men.


Central control of song in the canary, Serinus canarius. We have traced central nervous pathways controlling bird song in the canary using a combination of behavioral and anatomical techniques. Unilateral electrolytic brain lesions were made in adult male canaries whose song had been previously recorded and analysed on a sound spectrograph. After several days of postoperative recording, the birds were sacrificed and their brains processed histologically for degeneration staining with the Fink-Heimer technique. Although large lesions in the neostriatum and rostral hyperstriatum had no effect on song, severe song deficits followed damage to a discrete large-celled area in the caudal hyperstriatum ventrale (HVc). Degenerating fibers were traced from this region to two other discrete nuclei in the forebrain: one in the parolfactory lobe (area X, a teardrop-shaped small-celled nucleus); and a round large-celled nucleus in the archistriatum (RA). Unilateral lesion of X had no effect on song; lesions of RA, however, caused severe song deficits. Degenerating fibers from RA joined the occipitomesencephalic tract and had widespread ipsilateral projections to the thalamus, nucleus intercollicularis of the midbrain, reticular formation, and medulla. It is of particular interest that direct connections were found onto the cells of the motor nucleus innervating the syrinx, the organ of song production. Unilateral lesions of n. intercollicularis (previously implicated in the control of vocal behavior) had little effect on song. One bilateral lesion of HVc resulted in permanent (9 months) and complete elimination of the audible  components of song, although the bird assumed the posture and movements typical of song. Preliminary data suggest that lesions of the left hemisphere result in greater deficits than lesions of the right one. This finding is consistent with earlier reports that the left syrinx controls the majority of song components. Results reported here suggest a localization of vocal control in the canary brain with an overlying left hemispheric dominance.


Hormone concentrating cells in vocal control and other areas of the brain of the zebra finch ( Poephila guttata). Using the autoradiographic method in the zebra finch (poephila guttata), areas of the brain were identified which contain cells which accumulate testosterone. Among these areas are the caudal nucleus of the hyperstriatum ventrale, nucleus intercollicularis of the midbrain, and the tracheosyringeal portion of the hypoglossal nucleus of the medulla (nXIIts). These three are known to control or influence androgen dependent song and other vocalizations of passeriform birds, and nXIIts is composed of the motoneurons innervating the vocal (syringeal) muscles. Other areas containing hormone-concentrating cells are the medial preoptic area, nucleus periventricularis magnocellularis of the hypothalamus, dorsal infundibular layers, dorsomedial thalamus, lateral septum, magnocellular nucleus of the anterior neostriatum, periventricular medial neostriatum, nucleus taeniae of the archistriatum, and ventral paleostriatum augmentatum. Accumulation by cells in the preoptic area, hypothalamus, and limbic forebrain is consistent with a general vertebrate pattern of distribution of brain cells which accumulate sex steroids. Some of these same areas may be involved in the control of androgen dependent events such as courtship, copulation, aggression, and feedback regulation of the hypophysis.


Effects on visual tracking of delta9-tetrahydrocannabinol and pentobarbital. Detal-9-tetrahydrocannabinol and pentobarbital both interfered with visual tracking performance. Of these, delta9THC had a statistically significant effect (P less than 0.04) when the first 12 responses for each subject were analyzed at each half-hourly period.


Gustatory pathways in the bullhead catfish. 1. Connections of the anterior ganglion. The central projections of the external gustatory system in bullhead catfish were examined using orthograde degeneration and retrograde transport of horseradish peroxidase (HRP) techniques. Both large and small cells were observed in the anterior ganglion which contains a mixture of elements from the trigeminal, facial and anterior lateral line nerves. Some of the large cells on the lateral margin of the ganglion were found to belong to the lateral line system. No separation of trigeminal and facial nerve connections could be made. Using HRP, the relation between the barbels and specific ganglion regions was determined. The dorsalmost portion of the ganglion received recurrens nerve inputs (from taste buds on the trunk); the rostromedial portion of the ganglion, from the maxillary and mandibular barbel nerves. The facial lobe (similar to part of the n. solitarius) was found to be divided into lobules by fascicles of nerve fibers. The lateral lobule received input only from the dorsal-most part of the ganglion (recurrens nerve: trunk receptors); the intermediate lobule from the rostro-lateral part of the ganglion (nasal barbel); and the medial lobule from the ventral areas of the ganglion (maxillary and mandibular barbels). Thus a topographical relationship exists between the different taste receptor groups and their locus of representation in the facial lobe. The trunk receptors connect to the lateral lobule; the nasal barbel receptors to the intermediate lobule, and the maxillo-mandibular receptors to the medial lobule.


Retinofugal pathways in the lingnose gar Lepisosteus osseus (linnaeus). Retinal projections were studied with autoradiographic and silver methods in the gar, Lepisosteus osseus, one of the two surviving members of the holostean actinopterygians. Contralaterally, the retina projects to the preoptic nucleus of the hypothalamus, and, via the medial optic tract, to the dorsal thalamus, medial ventral thalamic nucleus, nucleus pretectalis profundus pars ventralis and pars dorsalis, and the medial portion of the deep layers of the central zone in the optic tectum. The dorsal optic tract projects to the lateral ventral thalamic nucleus, nucleus pretectalis centralis, and the superficial white and gray zone of the optic tectum. The ventral optic tract terminates in the nucleus of the ventral optic tract, the lateral and medial ventral thalamic nuclei, nucleus pretectalis superficialis, nucleus pretectalis centralis, nucleus pretectalis profundus pars ventralis, the basal optic nucleus, and the superficial white and gray zone of the optic tectum. Ipsilateral projections are to similar sites, except for an absence of inputs to the lateral ventral thalamic nucleus from the dorsal tract and to the nucleus pretectalis superficialis, nucleus pretectalis profundus pars ventralis, and the basal optic nucleus from the ventral tract. The presence of ipsilateral retinal projections in gars is compared to their presumed absence in teleosts, and comparisons of retinorecipient targets in gars are made with teleosts and with non-actinopterygian vertebrates.


An experimental inquiry into the central source of preganglionic fibers to the chick ciliary ganglion. Degenerative changes in the entire third nerve nucleus were studied following selective removal of the ciliary ganglion, usually of the right side, in day-old chicks. The operated animals were allowed to survive three, six, nine, and twelve days after ciliary ganglionectomy. Retrogarde degenerative changes proceed rapidly between three and nine days post-operatively, and are strictly limited to the ipsilateral accessory oculomotor nucleus. The changes are at first observed in the medial division of the accessory oculomotor nucleus, followed by the lateral division of the accessory oculomotor nucleus. By nine days following the operation, both the divisions of the accessory oculomotor nucleus are virtually depleted of almost all neurons. The other components of the oculomotor complex were not affected at any stage by ciliary ganglionectomy. These observations provide conclusive evidence that the accessory oculomotor nucleus is indeed the avian homologue of the Edinger-Westphal nucleus.


Functional role of efferents to the avian retina. I. Analysis of retinal ganglion cell receptive fields. Receptive fields of retinal ganglion cells were analyzed during extracellular microelectrode recordings in the optic tract of the lightly anesthetized pigeon. Four major types of receptive field can be distinguished among the 359 fibers studied. Twenty-five percent of the receptive fields are relatively simple, responding at on and at off to stationary spots of light in the central region. All of the receptive fields have inhibitory surrounds of varying strength that do not produce a response when illuminated alone, but antagonize responses from the central region. Motion sensitive units comprise 15% of the recorded population; they are similar to the on-off center type except that responses to stationary stimuli are absent or very weak while responses to moving stimuli are virorous. Directionally selective units also have the basic features of on-off, inhibitory surround cells, but respond to moving stimuli well from the preferred direction and not at all from the null direction. Directional cells have a broad range of null directions; in about one-third of the units the range becomes broader when the stimulus involves both center and surround of the receptive field, thus enhancing directional selectivity. Directionally selective units are common, comprising 38% of the units studied. Cells unresponsive to stimuli moving from anterior in the visual field are much more common than other types, while cells unresponsive to stimuli from posterior in the field are rare. A few units (11%) respond only at on or at off to stationary stimuli in their receptive field centers; they also have antagonistic but unresponsive receptive field surrounds. The area of the visual field sampled is uniform in regard to the relative numbers of the four major receptive field types.


Functional role of efferents to the avian retina. II. Effects of reversible cooling of the isthmo-optic nucleus. Efferents to the retina in the bird arise in the isthmo-optic nucleus of the caudal midbrain, and terminate on amacrine cells in the retina. The functional role of these efferents was studied by determining the receptive field properties of 107 optic tract fibers in the lightly anesthetized adult pigeon, and quantitating their responses to specific moving stimuli. While the recording from these fibers continued, the isthmo-optic nucleus was cooled by a thermoelectric cooling probe, and the response properties of the cells redetermined. Recording was maintained in half of the units long enough to observe recovery from cooling, and in several units the entire procedure was repeated. In 77 of the 107 units, responsiveness to all stimuli was decreased by removing efferent influences, whereas specific receptive field properties such as motion sensitivity or directionaltiy were not altered. All of the major receptive field types were affected in a similar fashion, irrespective of their position in the visual field. Responses to stimuli that did not involve the antagonistic surround were similarly affected by removal of the efferents, as were units were both weak and strong antagonistic surrounds. Efferents exert their influence on retinal ganglion cells by way of the amacrine cells on which they terminate. Data available on amacrines in Necturus indicates that they are inhibitory to ganglion cells. If amacrines have a similar role in the pigeon, then it may be stated that decreased activity in the centrifugal fibers leads to enhanced inhibition throughout the receptive fields of ganglion cells, and increased activity in the efferents produces disinhibition.


Taste pathways to hypothalamus and amygdala. The projections of a third order gustatory relay in the dorsal pons of rats have been traced using tritiated proline autoradiography and antidromic activation of pontine neurons from electrodes in the thalamus and amygdala. Labelled axons collect in the central tegmental tract and ascend to the thalamic taste area in the medial extension of the ventrobasal complex. The majority of the fibers remain ipsilateral, but a few cross in the rostral pons and midbrain. The largest crossing occurs at the level of the thalamic termination. Many fascicles of fibers continue rostrally by passing beneath the thalamic taste area, piercing the medial lemniscus, and spreading out along the dorsomedial corner of the internal capsule (IC). The terminal field at this level caps IC from the subthalamic nucleus down into the far-lateral hypothalamus. Labelled axons grandually penetrate through the internal capsule, and ramify throughout the underlying substantia innominata. This terminal zone extends laterally into the rostral end of the central nucleus of the amygdala, which is densely labelled to its caudal exremity. At the caudal end of the amygdala labelled fibers are visible in one component of the stria terminalis. These fibers can be followed over the dorsal thalamus into a smaller, but equally dense terminal area in the dorsolateral bed nucleus of the stria terminalis. The electrophysiological data demonstrate that pontine gustatory units can be antidromically activated by electrodes located in or near the central nucleus of the amygdala. Since many of the same units can also be driven from the thalamic taste area, at least some of the axons traced autoradiographically probably convey gustatory information to the hypothalamus and amygdala.


Fiber trajectories of olfactory bulb efferents in the hamster. Olfactory bulb efferents sweep caudally over the surface of the piriform lobe in a broad fiber sheet. The internal organization of this axon population was analysed by topologically transforming the cortical surface from its in situ cylindrical form into an unrolled (flattened) map. The distribution of degeneration elicited by restricted bulb lesions and fiber transections was then reconstructed onto this map. Most of the projection cortex of the main olfactory bulb is innervated in a widespread, non-topographic manner by axons that collect in the compact bundle of the lateral olfactory tract (LOT). LOT collateral branches bound for the prepiriform cortex veer laterally off the main trajectory of the tract at an angle of 50 degrees or less. Thus, transection of discrete fiber populations leaves only a small wedge-shaped pocket of totally denervated cortex distal to the cut. The medial half of the olfactory tubercle and the hippocampal rudiment receive their bulbar input along medially disposed fibers that do not join the LOT proper. The lateral half of the olfactory tubercle, however, receives an input from LOT fiber collaterals as well as these medial bulb efferents. Finally, much of the corticomedial amygdaloid complex receives fibers from the accessory olfactory bulb along a specialized subdivision of the LOT, the accessory olfactory tract. These observations are expressed in a schematic summary of the trajectories of olfactory bulb efferents as they appear in the unrolled map and in the more standard ventral view of the hamster brain.


Neuroplasticity in the rearrangement of olfactory tract fibers after neonatal transection in hamsters. Olfactory bulb efferent axons run caudally in the lateral olfactory tract (LOT) to end in a broad cortical field in the ventral forebrain. Principles governing the plastic rearrangement of this fiber population after early lesions were probed by cutting the tract in hamster pups and studying the distribution of surviving olfactory bulb projections in adulthood using silver and autoradiographic techniques. The spatial pattern of rearrangement proved to depend on the extent of the cut and also the age at which it was made. For example, after complete LOT section at seven days of age no bulb efferents appeared distal to the cut but the proximal projection increased in laminar thickness and spread laterally and medially beyond its normal cytoarchitectonic boundaries. This spread was less pronounced in animals with earlier or later lesions. After transection of only part of the LOT fibers at seven days of age the proximal input was similarly increased. Just distal to the transection, uncut fibers sprouted collaterals to fill the terminal sites vacated as a result of the lesion. In these cases, however, the farthest distal parts of the projection field lost their normal innervation. In a tentative interpretation of these data it is proposed that developing LOT fibers tend to conserve their total amount of axonal arbor. That is, when distal branches are pruned off surgically, the axon compensates by producing extra proximal branches. When an overabundance of proximal collaterals are produced in axons that have not been surgically pruned the growth of more distal axonal branches is stunted in compensation.


Synapses in the interpeduncular nucleus: electron microscopy of normal and habenula lesioned rats. Four types of synapses have been identified in the rat interpeduncular nucleus. The S synapses were formed by the mainhorizontal plexus of habenulointerpeduncular axons. They contained spherical synaptic vesicles and formed asymmetrical en passant contacts with dendritic processes. The crest synapses had all of these features in common with the S synapses, but occurred in pairs with the two contact zones coextensive and largely parallel on opposite sides of markedly attenuated dendritic processes. Synaptic glomeruli found in this nucleus consisted of several crest synapses engulfed by sheet-like dendritic processes. Identical crest synapses also occurred on dendritic crests. In a limited number of cases, both S and crest synapses arose in common from single axons. Following destruction of the habenular nuclei unilaterally or bilaterally, as expected from the above observations, both S and crest synapses underwent dense degeneration. In some cases with bilateral lesions and in all cases with unilateral lesions, only one of the two axonal endings forming a crest synapse degenerated while the other remained unaffected. Axons containing flat vesicles forming symmetrical axodendritic synapses, axosomatic synapses, and subjunctional bodies associated with S or crest synapses were all minor features of this nucleus. Aspects of the unusual synaptology of this nucleus are discussed in terms of morphology, physiology and transmitter chemistry.


Organization of retinocollicular pathways in the cat. A direct pathway from the temporal retina to the contralateral superior colliculus has been demonstrated using anterograde degeneration methods. This observation of a crossed pathway to the most rostral pole of the colliculus confirms electrophysiological evidence that within each colliculus the whole eye, rather than just its hemiretina is represented. Autoradiographic tracing methods have been used to study the total tectal projection of each retina. Following intreocular injections of 3H proline, there is dense label in most of the superficial gray of the contralateral colliculus, but in this region there are several small areas of only relatively sparse label. In contrast, the superficial gray of the ipsilateral colliculus contains islands or patches of transported label. These patches are present throughout an extensive region of the colliculus and suggest that the ipsilateral retinocollicular pathway in the cat may be more substantial than earlier reports have indicated. There is no ipsilateral label within the most caudal parts of the colliculus which receives afferents from the monocular crescent of the rostral pole is free of ipsilateral input, and we suggest that this corresponds to the region receiving the crossed input from the temporal retina.


Increases in collateral axonal growth rostral to a thoracic hemisection in neonatal and weanling rat. The spinal cords of newborn and weanling rats were hemisected at the mid-thoracic level. Control studies revealed that Fink-Heimer positive debris was absent in the gray matter at three months postoperative. The remaining animals were given a second lesion, a high cervical spinal hemisection, at five to seven months after the original thoracic hemisection. The pattern of degeneration rostal to the thoracic lesion was compared with similar regions of the spinal cord from animals receiving only a cervical hemisection at the adult stage. In neither experimental group of doubly hemisected rats was there any degeneration observed below the thoracic lesion site, even though no glial or connective tissue scar had formed in animals originally operated at birth. Thus no regeneration had occurred. At least one segment above the initial hemisection: 1. the majority of degenerating axons were localized toward the lateral edge of the spinal cord, especially in the doubly lesioned neonatal group; 2. the erae of ipsilateral white matter was reduced more in the neonatal than the weanling operates; 3. there was an upward shift in axonal diameter of ipsilateral fibers in both the region of the rubrospinal tract and the ventrolateral portion of the lateral funiculus of the doubly hemisected rats when compared with the cervically lesioned controls; 4. a significantly greater amount of degeneration was present in lamina VII of Rexed in both the neonatal and weanling experimental operates (p less than 0.05 weanling; p less than 0.001 neonate); 5. no mean difference in area was seen between the ipsilateral and contralateral gray matter in any group for the segments of the spinal cord in which the judgements and measurements were taken. These data suggest that there has been sprouting of axons from descending nerve tracts rostral to the thoracic lesion in both the neonatal and weanling experimental groups. The question remains whether the sprouting of descending nerve tracts is from collateral of axons which normally project rostral to the thoracic hemisection and are not cut by the thoracic lesion (collateral sprouting) or from collaterals of lesioned axons (regenerative sprouting). Present evidence favors collateral sprouting, expecially in the neonatal operate where much retrograde cell death appears to have taken place.


Changes in the magnocellular portion of the red nucleus following thoracic hemisection in the neonatal and adult rat. The spinal cords of newborn (0-3 day old) and adult rats were mid-thoracically hemisected. Ninety days later a glial and connective tissue scar had formed at the lesion site in the adult hemisected rats while in neonatally lesioned animals only normal appearing regions of the contralateral spinal cord were found in the area of hemisection. Comparisons of the magnocellular portions of the red nucleus (MPRN) revealed a decrease in cell number in the MPRN contralateral (C-MPRN) to the spinal lesion. However, only in the newborn operates was there massive cell loss accompanied by reduction in area and change in shape of the nucleus. These changes were most obvious in the caudal and ventrolateral portions of the C-MPRN. Pooled data from each group of operates indicated that significantly more cells were lost in the C-MPRN in the newborn than in the adult operates (p less than 0.01). Neurons of the C-MPRN which are known to project to the lower cervical and upper thoracic segments of the spinal cord (Brown, '74; Gwyn, '71) remained undamaged after the mid-thoracic hemisection in both groups. However, neurons of this region were enlarged in both groups when compared to a similar region of the ipsilateral MPRN. These neurons were found to be more enlarged in the newborn than in the adult operates (p less than 0.01). This result indicates that massive retrograde cell death takes place after a mid-thoracic hemisection in the neonatal rat. The retrograde degeneration of axotomized neurons partially may explain why CNS regeneration is not found in the immature mammal even though many of the factors thought to limit regeneration in the adult mammal may not be apparent. The increase in cell size of C-MPRN neurons which remain in the neonatal animals after mid-thoracic hemisection may be related to the increase in axonal size found in the region of the rubrospinal tract rostral to the thoracic lesion reported earlier (Prendergast and Stelzner, '76a). Both the increase in axonal and perikaryal size are hypothesized to be related to the increased distribution of supraspinal axons found in the gray matter rostral to a hemisection of the neonatal rat spinal cord.


A somatotopic and functional representation of the respiratory muscles in the trigeminal and facial motor nuclei of the carp (Cyprinus carpio L.). The place of the cells innervating six most important respiratory muscles in the carp (Cyprinus carpio L.) were determined by means of the horseradish peroxidase technique. It was found that these muscles are represented in the rostral and caudal subdivisions of the trigeminal and facial motor nuclei. All connections were ipsilateral. The distribution of the various cell populations over the different subnuclei has been related to the function of the respective muscles in the respiratory cycle. On these grounds it could be concluded that muscles active in the contraction phase of respiration are represented in the rostral subnuclei, the expansion muscles in the caudal subnuclei. This arrangement appears to be a compromise between a somatotopic and functional and functional organization.


Postnatal maturation of rat Purkinje cells cultivated in the absence of two afferent systems: an ultrastructural study. Organized cultures of newborn rat cerebellum were established in Maximow chambers in order to study the maturation of Purkinje cells in absence of afferent systems. In the first model, standard cultures were devoid of extracerebellar afferents mossy and climbing fibers. Despite this absence, somatic spines appeared upon Purkinje cells during the first week in vitro and maturation proceeded normally except for the almost absence of spiny branchlets. Large dendritic trunks were studded with numerous spines, some of which were naked, a few bearing isolated post-synaptic densities and others occupied by boutons of parallel fibers. Stellate and basket axons made synapses upon the smooth portions of dendrites and soma. In a second model, the cultures were fed the antimitotic drug methylazoxymethanol (MAM) to prevent multiplication of granule cell precursors. Despite the absence of climbing and parallel fibers, the elongation of Purkinje dendrites was not prevented, but again the dendritic arbor consisted of large trunks studded with spines; somatic as well as dendritic spines were contacted by large boutons identified as Purkinje recurrent collaterals (PRC). It is concluded that the Purkinje cell possesses a large autonomy from afferent systems as to the growth of soma and dendrites. Conversely, the geometry of the dendrite and especially the spiny branchlets depend on the presence of both climbing and parallel fibers. One may conclude from the above experiments that specificity of synaptic contacts is maintained as long as postsynaptic sites are not devoid of their normal afferents. Heterologous synapses are formed when postsynaptic sites are present, their normal afferents absent and aberrant ones increasing by collateral sprouting. Such is probably the case in the second model of this study.


Neurogenesis in the visual system of the rat. An autoradiographic investigation. Rats of the BD III strain were injected with a single dose of 3H-thymidine on either the twelfth, fourteenth, sixteenth, eighteenth or twentieth day of gestation (ED 12. . . . .ED 20) or on the postnatal day one, three, or seven. Animals were killed at age 22 to 24 days. DNA synthesis, as an indicator of cell division, was studied in matrix precursors of nerve and glial cells in the visula centers, including the lateral geniculate body (LGB), the superior colliculus (SC) and the visual cortex (VC). It was found that proliferation of matrix precursors of nerve cells destined for all the regions studied was in progress on ED 12. In the subcortical regions (LGB, SC) this process was substantially more advanced than in the VC. The first neuroblasts appeared in the SC (ED 12) and only later (ED 14) in the LGB and VC. In comparison with the LGB, VC neuroblasts were quite rare on ED 14 and were present only in layer VI. They appeared more frequently in this region only after injection of isotope on ED 16. Matrix cell proliferation and nerve cell formation ceased in the LGB between ED 16 and ED 18. The number of labeled cells arising after injection of the isotope on ED 16 indicates that neurogenesis ceased somewhat earlier in the dorsal nucleus of the LGB than in the ventral. In the SC the last neurons arose between ED 18 and ED 20, and in the VC, with the possible exception of a few granular neurons (which may continue division into the first few days postnatally), proliferation continued until the end of gestation. The origin of neuroblasts initially followed a caudo-rostral gradient. Later, the times of neurogenesis in the regions studied overlapped significantly. This is clear, for example. on ED 16, when neurogenesis in the mesencephalic SC continued for about two days longer than in the more postral LGB, and coincided with that in the VC, especially in the deep layers. The end of neurogenesis in the LGB, especially in the ventral nucleus, coincided with the time of neurogenesis in the deep cortical layers. In the VC, and partly also in the SC, an inside-out pattern of proliferation and neuron formation was confirmed. The times of proliferation of precursor cells, with the exception of the very end of neurogenesis, substantially overlapped within both these regions. The degree of this overlapping, described in terms of Labeling Index values, decreased towards the end of the neurogenetic period. Division of neuroglial cell precursors, started as early as on ED 14 in/for subcortical centers (LGB, SC), but not until ED 18 in/for the VC. A few labeled endothelial-like cells were observed in all regions studied after isotope injection on ED 12.


Afferents to the cerebellar lateral nucleus. Evidence from retrograde transport of horseradish peroxidase after pressure injections through micropipettes. HRP was injected by pressure from glass capillary micropipettes unilaterally into the lateral nucleus of rat so as to encompass the entire nucleus, but without spread into the interpositus nuclei. The cells of origin of the afferents to the lateral nucleus were studied after retrograde transport of the HRP. The reticulotegmental nucleus of the pons was labelled bilaterally and is the major source of crossed and uncrossed reticular imputs. The pontine nuclei also provide extensive crossed and uncrossed afferents. The inferior olive gives a large crossed olivo-lateral nucleus projection and a minor uncrossed input. The trigeminal nuclear complex--the nucleus of the spinal tract and the mesencephalic, principal sensory, and motor nuclei--all provide uncrossed afferents. The rostral portion of the lateral reticular nucleus gives a small crossed and uncrossed projection while the perihypoglossal nuclei and the dorsal parabrachial body give crossed afferents to the lateral nucleus. The norepinephrine afferent system from the locus coeruleus is represented by one or two heavily labelled cells and the serotonin raphe systems come from at least five raphe subgroups, the dorsal, superior centralis, pontis, obscurus and magnus nuclei. No evidence was found for commissural fibers between ipsilateral or contralateral cerebellar nuclei, or afferent axons from the spinocerebellar nuclei and the paramedian retricular nucleus. The significance of these sources of afferent imputs to the lateral cerebellar nucleus is discussed. The question is raised of the direct relationship between size of terminal axonal arborization and the quantity of HRP granules present in a cell retrograde transport. The limitations of the HRP method for detecting subtle local differences in the distribution of afferents within the heterogeneous groups of neurons in the lateral nucleus are discussed.


On the striate area of the human isocortex. A Golgi- and pigmentarchitectonic study. The distribution pattern of the pigment in various layers of the primary visual cortex of man is described by means of a method for the stereomicroscopical demonstration of neurolipofuscins in sections up to 1,000 mum thick. This paper presents the striate area as being composed of 12 cellular layers, which can be clearly delineated by a successive examination of both the Nissland the pigment preparations. Layers I, II, and III resemble more or less the corresponding laminae of other isocortical fields. The pyramids within the deep portion of layer III are unusually small and additionally are intermingled with thorny stellate cells which are a characteristic component of layer IV. Accordingly, this band of cells is called IIIc-IVa. The subjacent layers IVb and IVcalpha are poorly pigmented. In Nissl-preparations layer IVb, which contains only a few cells, is followed by a broad band of cells tightly packed together, which includes layers IVcalpha, IVcbeta, IVd, and Va. These layers cannot be reliably delineated in Nissl-preparations but appear in pigment preparations as distinct and sharply limited laminae. Particularly, layer IVcbeta, which is built up by minute thorny stellate cells, is marked by coarse and intensely stained lipofuscin granules, whereas IVd is mainly populated by minute pyramids which are almost devoid of pigment deposits. A remarkably narrowed layer Va, the pyramids of which store a medium number of lipofuscin grains, completes this series (IVcalpha, IVcbeta, IVd, Va). The cell-sparse layer Vb is almost devoid of pigment granules. Layer VIa has specialized pyramidal cells containing finely grained pigment. The relatively broad layer VIb shows loosely arranged cells poorly endowed with lipofuscin granules.


Cell growth in the lateral geniculate nucleus of kittens following the opening or closing of one eye. Closing one eye of kittens at 23 days after birth resulted in paler Nissl staining of the deprived cells in the lateral geniculate nucleus (LGN) that was detectable two days later. Differences between the two sides of the brain in mean cell area increased to a peak in binocular lamina A at 4--6 days after eyelid suture, and then fell to a trough at eight days before rising progressively to a higher level at 31 days. In lamina A1 the peak and trough were later. Opening one eye of kittens after 23 days of binocular closure from birth resulted in more intense Nissl staining of the stimulated cells in the LGN that was detectable four days later. The stimulated cells grew faster than the cells connected to the eye that remained closed, and this differential growth reached a peak at 17 days in binocular lamina A and at 21 days in lamina A1 before falling to a trough at 26--31 days. These results are compared with the time course of anabolic changes that have been measured in other neurons during stimulation. The coefficient of variation of cell size was computed and found to be slightly decreased for deprived cells and increased for stimulated cells. This suggests that larger cells changes their sizes proportionately more than smaller cells. The cells were measured in fronzen sections without shrinkage, and the areas are at least 25% larger than those reported previously after paraffin or celloidin embedding.


Efferent connections of the caudate nucleus in the Virginia opossum. The efferents of the opossum's caudate nucleus were investigated by charting the the fiber degenerations produced by electrolytic lesions in various parts of this nucleus. By the aid of a modified Fink-Heimer procedure, degenerating fibers were traced from each of the lesions to the small globus pallidus in which they appeared to be distributed in an orderly dorsoventral pattern. Fibers from all lesion sites in the caudate nucleus were found to terminate in the entopedunucular nucleus. In the substantia nigra, caudatofugal-fiber degeneration was confined in all cases to the rostal part of the pars reticulata, and was densest in the medial one-half of this nucleus. Only from lesions in the ventromedial part of the caudate nucleus could degenerating fibers be traced to the nucleus ansae lenticularis. No fiber degeneration could be traced rostrally from the lesions, or to the putamen, red nucleus, subthalamic nucleus, or pons.


The effect of frequency of feeding on milk secretion in the Ayrshire cow. Four Ayrshire cows in their second lactation were used in a 4 X 4 Latin square experiment to determine the effects of frequency of feeding on milk secretion. The animals were given a diet consisting of hay, sugar-beet pulp and dairy concentrates (34-9:19-7:45-5 on a dry-matter basis) either in 2 equal meals/d at 07.00 and 17.00 h or with the hay in 4 meals at 07.00, 12.00, 17.00 and 22.00 h and the sugar-beet pulp and concentrates in 24-h meals. Each feeding frequency was tested at 2 levels of feeding providing either 100 or 80% of the standard requirements for energy. There were no significant effects of frequency of feeding on milk yield or on the milk content of fat, protein, lactose, solids-not-fat or total solids, but underfeeding depressed the yield of milk and of all milk constituents. Neither the frequency nor level of feeding had appreciable effects on the fatty-acid composition of the milk fat or on the blood glucose concentration, but in animals fed frequently plasma urea levels were lower than in animals fed twice daily and there was no postprandial peak in urea concentration.


Formation of acetaldehyde from threonine by lactic acid bacteria. Group N streptococci were found to cleave threonine to form acetaldehyde and glycine. Threonine aldolase, the enzyme catalysing this reaction, was found in all strains except Streptococcus cremoris Z8, an organism which had been shown previously to have a nutritional requirement for glycine. The enzyme was strongly inhibited by glycine and cysteine. The inhibition showed characteristics of allosteric inhibition and was pH-dependent. Inhibition by glycine, but not by cysteine, was highly specific. Analogues and derivatives of cysteine which contained a thiol group and a free amino group inhibited the activity of threonine aldolase. The presence of a carboxyl group was not necessary for inhibition. The cleavage of threonine by whole-cell suspensions was stimulated by either an energy source to aid transport or by rendering the cells permeable to substrate with oleate. Threonine did not appear to be degraded by enzymes other than threonine aldolase, as threonine dehydratase activity was low and NAD- and NADP-dependent threonine dehydrogenases were absent.


A rumen linear programming model for evaluation of concepts of rumen microbial function. A linear programming model provides for analysis of general input-output relationships in the rumen, for evaluation of competitive relationships among rumen microbes, and for computation of optimal relationships in the rumen. Eight rumen microbial groups defined on the bases of substrate specificity, nutrient requirements for growth, fermentation products, and relative metabolic activities comprise the central core of the model. Relative metabolic rates of microbial groups calculated from their cell sized were used as coefficients in the objective function. The model was used to evaluate effects of different amounts of protein from feed and various carbohydrates upon microbial population and fermentation patterns as accommodated by current concepts. During the several solutions of the model, considerable simplification of the rumen microflora occurred. This implies that current data and concepts, and the hypothesis regarding relative metabolic rate, as represented in the model, do not accommodate adequately competitions among the several rumen microbial species and, thus, that additional data and concepts regarding rumen microbial interactions are required. Also evaluated were effects of ingestion of bacteria by protozoa upon over-all rumen function, absolute microbial cell yields, cell yields per mole of adenosine triphosphate, and factors affecting these.


Growth and morphology of Streptococcus bovis and of mixed rumen bacteria in the presence of aflatoxin B1, in vitro. The metabolism of aflatoxin B1 to other fluorescent compounds was studied in rumen organisms. No fluorescent metabolites were found by chromatographic procedures which separate aflatoxin B1 from them. Rumen bacteria (mixtures and Streptococcus bovis) incubated with aflatoxin B1 grew in long chains whereas cells in control cultures were mainly doublets with some short chains. Growth of mixed rumen bacteria and Streptococcus bovis cultures was inhibited by aflatoxin B1 as measured by change in optical density and by incorporation of [carbon-14] adenine into nucleic acids.


Effect of sampling location, time, and method of concentration of ammonia nitrogen in rumen fluid. In experiment 1, rumen fluid samples were obtained from dorsal, midpoint, and ventral regions of the rumen of rumenfistulated dairy cows at hourly intervals for 24 h. Dorsal, midpoint, and ventral rumen fluid samples contained 16.1, 14.2, and 12.1 mg/100 ml of ammonia nitrogen. Time after feeding resulted in differences in the concentration of ammonia in rumen fluid with the greatest concentration 30 to 90 min postfeeding. The concentration of ammonia nitrogen was greater for cows fed a high-concentrate diet (14.7% crude protein, 86% concentrate) than for cows receiving a normal diet (15.1% crude protein, 42% concentrate), 15.2 versus 13.1 mg/100 ml. In a second experiment, rumen fluid samples were obtained immediately before feeding and at 1 and 6 h postfeeding by stomach tube and from dorsal, midpoint, ventral, and mixed rumen contents via a rumen cannula. Concentrations of ammonia nitrogen (mg/100 ml) in the respective rumen fluid samples were 14.3, 23.0, 18.3, 17.2, and 19.6. Location of sampling, method of sampling, time of sampling, type of diet, and rumen fluid volume are factors that affect the concentration of ammonia nitrogen in rumen fluid.


Hexachlorobenzene retention and excretion by dairy cows. Two groups of three cows each were fed either 5 or 25 mg of hexachlorobenzene per day for 60 days. A reference compound, DDE (1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene), was fed to the cows at the same rate. Residues were determined in milk at 5-day intervals during the 60-day dosing period and for 60 days after dosing was stopped. The concentration of hexachlorobenzene in milk fat increased more slowly than did that of the reference compound during feeding. The ratio of the two was 1:2 at 10 days, but approached 1:1 after 40 days of feeding. The average concentrations in milk fat for the 40th to 60th days of feeding were 9.0 and 10.4 ppm with the 25-mg/day intake and 2.1 ppm for each compound with the 5-mg/day intake. Corresponding values in subcutaneous body fat were 8.8, 8.0, 1.9, and 1.4 ppm at 60 days. Milk fat concentrations of hexachlorobenzene declined 32% and those of the reference 51% within 15 days after feeding stopped. Thereafter, the decline was slower but similar for both compounds. Biological half-lives ranged from 29 to 64 days for individual cows. The milk fat:body fat concentration ratio was .87:1 for hexachlorobenzene and .71:1 for the reference during the 60 days when cows were not fed the compounds.


Effects of adrenalectomy upon ruminant liver and mammary function during lactation. Adrenalectomized, lactating ewes treated with a mineral corticoid or a mineral corticoid plus cortisol and sham operated ewes nursing one or two lambs were in a study of physiological role(s) of glucocorticoids in lactating ruminants. Measurements of ewe and lamb weight changes; feed intake; milk yield and composition; blood potassium; metabolite oxidation rates by mammary, liver, and kidney tissue slices; glucose production in liver and kidney tissue slices; and enzyme activities in mammary and liver tissue. 1) Mammary metabolism and milk biosynthesis in ruminants are not strongly dependent on glucocorticoid. 2) Liver gluconeogenesis in ruminants appears to be regulated in part by glucocorticover and kidney of ruminants may respond differently to adrenalectomy; thus, in ruminants, kidney may not be as good an indicator of liver gluconeogenic responses as it is in rodents. 4) Liver and mammary enzymes are not affected significantly by adrenalectomy or glucocorticoid therapy. 5) Glucocorticoids play a less significant role in the regulation of liver and mammary enzyme and metabolism in sheep than in rats.


Hormone induced lactation in the bovine III. Dynamics of injected and endogenous hormones. Three lactations were induced in two cows in two trials by twice daily subcutaneous injections for seven days. Each injection contained .05 mg/kg body weight of estradiol-17beta and .125 mg/kg body weight of progesterone. Included with the hormones in trial 1 was a combination of 233 muCi of [6,7-hydrogen-3] estradiol-17beta and 12 muCi of [4-carbon-14] progesterone. During trial 2,2050 muCi of [2,4,6,7 hydrogen-3] estradiol-17beta were incorporated with the hormones for one cow and 2344 muCi of [1,2,6,7 hydrogen-3] progesterone for the other. Radioactivity from hydrogen-3 and carbon-14 was detected in urine and feces but not in mammary secretion during trial 1. Isotope recovery from [hydrogen-3] estradiol-17beta was too low to quantify. By day 17, 90% of the carbon-14 from progesterone was accumulated in feces and 8.0% in urine. For trial 2, recoveries of radioactivity by day 28 from urine, feces, and mammary secretions were, respectively, 32.4, 56.2, and less than .002% from [hydrogen-3] estradiol-17beta and 6.0, 87.7, and .25% from [hydrogen-3] progesterone. Estrogen and progesterone in plasma averaged 1.83 and 2.61 ng/ml during days 2 to 7. Estrogen decreased more rapidly than progesterone post-treatment, but both had decreased by days 16 to 19 when average prolactin of plasma increased. Quantitative measures of estradiol 17-beta and progesterone absorption, whether in plasma or excreta, were correlated.


Mathematical modeling of the mastitis infection process. Mastitis infection is a biological process with a serial structure and stochastic in nature. A mathematical model for this process was developed from Markov chain theory. The unit of the process was an individual quarter; and four infection processes for first, second, third, fourth or higher lactation were described. Two Markov matrices which probabilistically determine the transitions between seven mutually exclusive states at each stage of the process were defined. After its validation, the model was used to determine the lactational consequences of the mastitis infection by calculating expected milk-yield productivity. Expected milk-yield productivity for an individual quarter was .93, .88, .85, and .84 for first, second, third, and fourth or higher lactation. The expected milk yield productivity of a quarter as a function of its state at the beginning of the process also was calculated. The quarters infected but not clinical have the lowest productivity.


Intracellular distribution of zinc and zinc-65 in calves receiving high but nontoxic amounts of zinc. The zinc homeostatic control breakdown in cattle fed a high but nontoxic amount of zinc was investigated. Liver copper was decreased by the 600 ppm added dietary zinc indicating altered copper metabolism. However, duodenal copper, liver and duodenal iron and manganese were not affected. Zinc-65 in blood was reduced 90% by the high dietary zinc 48 h following oral zinc-65 dosing. The 600 ppm supplemental zinc increased zinc by 500% in liver, 20 times in pancreas and kidney, and 100% in the duodenum. The increased liver and duodenal zinc was confined largely to the soluble cell fraction. This concentrating of excess tissue zinc in the soluble fraction may be an adaptive mechanism which detoxifies large quantities of zinc and prevents disruption of normal cellular activity.


Composition of the milks of dairy cattle. II. Ash, calcium, magnesium, and phosphorus. Milks from commercial dairy herds in Southeastern Pennsylvania were analyzed for ash, calcium, magnesium, and phosphorus. Milk samples were from single milkings of 151 healthy purebred cows in midlactation. The data represent market milk composition. Average values for all animals were ash .78%, calcium 1.25 g/liter, magnesium .11 g/liter, and phosphorus 1.14 g/liter. The data also were grouped and analyzed by breed and beta-lactoglobulin phenotype. Breeds differed in all inorganic components with 18 to 33 cows per breed. No differences in ash, calcium, magnesium, or phosphorus were significant when the data were grouped by beta-lactoglobulin phenotype.


Mammary transfer of vitamin A alcohol and ester in lactating dairy cows. The effect of intravenous injection of vitamin A alcohol and vitamin A ester on the vitamin A concentration of bovine milk was studied. Holstein cows received either an intravenous injection of 1 million international units (IU) of vitamin A alcohol, 1 million international units of vitamin A palmitate, or served as controls. Blood serum and milk were sampled at intervals prior to and following injection. Mean concentrations of vitamin A in milk at time 0 were 59.0, 52.0, and 58.4 mug/100 ml for the control, alcohol, and palmitate treatments. Mean contrations of vitamin A in milk, expressed as mug/100 ml or mug/g fat of cows receiving vitamin A alcohol at +195, +451, and +678 min postinjection, were greater than concentrations for either control cows or cows receiving palmitate. Partition of milk Vitamin A at +195 min postinjection for cows injected with vitamin A alcohol showed 4.1% of the recovered vitamin A in the alcohol and 95.9% in the ester form. Injection of vitamin A palmitate had little effect on milk vitamin A concentration. Vitamin A in serum (mug/100 ml) at 195 min after vitamin A alcohol injection was higher (100.4) than for either control (84.0) or cows injected with vitamin A palmitate (89.0).


Lactational response of dairy cows to oral administration of a synthetic glucocorticoid. Effects of oral administration of 0, 5, 10, or 20 mug of 6 alpha, 9 alpha-difluoro-16alpha-methyl-prednisolone (Flumethasone) daily on milk and milk component yields, udder health reproductive performance, and body weight change were measured. With 24 Holstein cows in a randomized block design, daily Flumethasone administration was initiated 4 days postpartum and continued until milk secretion ceased or the completion of 305 days on treatment. Mature equivalent yields (adjusted for days open) of milk, solids-not-fat, and milk fat were higher for cows receiving 5 or 10 mug Flumethasone than for cows receiving 0 or 20 mug Flumethasone daily. Lactations for cows receiving 20 mug Flumethasone were 37 to 48 days shorter than lactations for cows in other treatment groups. There were no differences between treatment groups for measures of udder health, reproductive efficiency, and body weight changes. A second experiment of 45 Holstein cows in their second or third lactations included treatments 0, 5, or 10 mug Flumethasone daily for 25 days followed by a 5 day withdrawal. Treatments were begun between day 210 and day 270 of lactation. During the 30 days, Flumethasone failed to alter milk production. These experiments indicate that Flumethasone given continuously increased milk components and milk yield but had no significant effect on milk yield when given only during the latter stage of lactation.


Effect of adrenalectomy on rat mammary metabolism. Rates and patterns of metabolism of specifically labeled glucose, pyruvate, and acetate by mammary slices indicated that rates of glucose metabolism through the Embden-Meyerhof and pentose phosphate pathways and relative pentose cycle activity were decreased by adrenalectomy. Fatty acid synthesis from acetate was not impaired. Capacity to provide reduced nicotinamide adenine dinucleotide phosphate for fatty acid synthesis was not the limiting factor responsible for decreased fatty acid synthesis from glucose in adrenalectomized, lactating rat mammary glands. Oxidation of [1-carbon-14]pyruvate and fatty acid synthesis from [2-carbon-14] pyruvate were not affected by adrenalectomy. Indirect reasoning based upon the data reported led to the suggestion that decreased fatty acid synthesis from glucose after adrenalectomy may have resulted from decreased formation of triose phosphate via the phosphofructose-kinase reaction. It is suggested further that most of the decreases in rat mammary enzyme after adrenalectomy have no direct (causative) relationship to depressed lactational performance after adrenalectomy. Restriction of food intake caused decreased fatty acid synthesis from glucose, pyruvate, and acetate, but relative fatty acid esterification activity was higher than in control and adrenalectomized animals. The effects of adrenalectomy on mammary gland metabolism can be dissociated partially from reduced food intake.


Factors influencing rumen microbial growth rates and yields: effects of urea and amino acids over time. Washed cell suspensions of mixed rumen bacteria were used to evaluate effects of 100% urea-nitrogen and 75% urea-nitrogen plus 25% amino acid-nitrogen in growth media upon microbial growth rate and yield, specific rate of glucose consumption, and incorporation of glucose into mixed cells, carbon dioxide, and end products. Rumen microbial dry matter, nitrogen, ribonucleic acid, deoxyribonucleic acid, glucose disappearance, and production of volatile fatty acids were considerably higher in medium containing urea plus amino acids as compared with urea only. Specific growth rates of microbes were .104 and .203 and mean doubling times were 6.7 and 3.4 h in the urea and urea plus amino acid growth media. Microbial growth in mg per 100 mg glucose used, per mole glucose and per mole adenosine triphosphate (ATP), and specific rate of glucose consumption in mmol per mg cells-h were 19.3, 34.7, 15.4, and .016 with urea, and 24.4, 44.2, 20.6, and .014 with urea plus amino acids. Percentages of catabolized glucose incorporated into microbial cells, carbon dioxide, and end products did not differ between treatments and averaged 19.5, 7.8, and 64.4%.


Dynamics of fermentation of a purified diet and microbial growth in the rumen. Ruminal fermentation and disappearance of glucose, starch, and cellulose, and incorporation of glucose and starch into microbial cells were estimated in a fistulated Jersey cow fed twice daily a purified diet containing urea as the sole nitrogen source. Estimated rumen volume was 59.8 liters. Turnover time and rate of passage of rumen contents were 33.4 h and 1.8 liters per h. Turnover times of glucose, starch, and cellulose were .17, 4.7, and 14.2 h. Fermentation times of glucose, starch, and cellulose were .17, 5.5, and 25.1 h. Percentages of glucose, starch, and cellulose utilized in the rumen were 99.4, 85.4, and 60.6. Thus, 18.5% of the carbohydrate fed bypassed rumen fermentation, and 81.5% was utilized in the rumen. All glucose disappeared from the rumen within an hour. An average of 32.1, 43.0, and 14%, respectively, of glucose utilized was incorporated into microbial cells, volatile fatty acids, and carbon dioxide. Percentage of starch incorporated into cells varied, with time being highest 2 h after feeding at 40% and lowest at 20%, 10 h after feeding. Respective percentages of starch incorporated into microbial cells, volatile fatty acids, and carbon dioxide were 32.4, 45.9; and 13.3. Total microbial protein and cell yields per kilogram carbohydrate utilized in the rumen were 77.1 and 117.5 g. Microbial cell yield per mole (estimated) of adenosine triphosphate was 16.2 g.


Factors influencing rumen microbial growth rates and yields: effect of amino acid additions to a purified diet with nitrogen from urea. Effects of isonitrogenous urea and amino acid additions upon microbial growth in rumen contents from a cow fed a purified diet in which urea was the sole nitrogen source were studied. Incorporation of amino acids into microbial cells, volatile fatty acids, and carbon dioxide was estimated. Rates of microbial growth, volatile fatty acid production, and effects of amino acids upon microbial nitrogen yields were highest right after feeding and decreased with time after feeding. Microbial growth and amounts of amino acids incorporated into microbial cells, volatile fatty acids and carbon dioxide were related closely to quantity of starch remaining in the rumen. High amounts of starch increased microbial protein synthesis from carbon-14 labeled amino acids and reduced amounts of amino acid fermentation. Estimated microbial protein yields per day were 326.0, 444.4, 497.3, and 527.3 g when 0, 15, 30, and 45 mg amino acid nitrogen replaced urea nitrogen during incubation. Respective values for microbial cells per mole estimated adenosine triphosphate were 15.2, 19.2, 21.0, and 24.5. Microbial cell yields per kg carbohydrate digested were 139.0, 189.5, 212.0, and 224.8 g for 0, 15, 30, and 45 mg amino acid nitrogen. Addition of small amounts of amino acids to a diet containing urea as the sole nitrogen source improved considerably rumen microbial protein yields.


Zinc absorption from the small intestine in young calves. Zinc-65 was injected directly into various sites throughout the small intestine of Holstein calves fed a low zinc diet. Zinc absorption was determined by comparing zinc-65 content of blood, liver, kidney, lung, heart, pancreas, rib, muscle, and other tissues. Tissue zinc-65 was plotted against site of injection expressed as percentage of intestinal length from proximal to distal ends. Zinc absorption occurred throughout the small intestine with the amount absorbed per centimeter of length about equal throughout the small intestine. This is in contrast to conclusions from studies of rats indicating more absorption in the proximal part of the small intestine. The difference appears to have been due to artifacts in the procedures with rats. However, species and dietary differences may have affected relative findings between this and earlier work.


Relationship of testicular weight to age and scrotal circumference of Holstein bulls. Testicular weights were recorded for 250 Holstein bulls ranging in age from 19 to 189 mo. The multiple regression equation that best described the relationship between paired-testes weight and age of bull was Y = -368.8 + 952.2 LOG X - 180.3 (log X)2, where Y is paired-testes weight in g and X is bull age in mo. Correlations between these two variables were .86 for 24 bulls less than or equal to 60 mo of age and .60 for 23 greater than 60 mo of age. A linear regression equation for 47 bulls was Y = -645.5 + 34.0 X, where Y is paired-testes weight in g and X is scrotal circumference in cm measured within 1 mo of slaughter. The correlation coefficient was .79. Differences in testicular weights of bulls of similar ages were large. These should be taken into account by measuring scrotal circumference and selecting young bulls with larger testes and superior spermatozoal producing capabilities.


Body weight changes in lactating purebred and crossbred dairy cattle. Body weights of 396 first and 205 second lactation cows in the first, second, third, and fourth generations of the dairy cattle crossbreeding project at the Illinois Agricultural Experiment Station were studied. Body weights were taken regularly on each animal during the first 9 mo of each lactation with the first weight being taken 3 to 4 days after parturition and subsequent weights at intervals of 30 days thereafter. Among least-squares means there was a decrease in body weight during the first 1 or 2 mo of lactation and a steady increase thereafter in all breed groups for both first and second lactations. There were differences between cows as well as between months for all breed groups in each generation for both lactations. Sums of squares for months were partitioned into linear, quadratic, and cubic components. A third degree polynomial was fitted for each breed group within each generation and separately for each lactation showing the changes in body weights.


Bovine plasma corticoids during parturition. Concentrations of adrenal corticoids in plasma were determined by a competitive protein-binding technique using jugular blood samples taken three times daily and more frequently during labor from 21 cows before and after parturition. The prepartum value was 6 ng/ml with a sharp rise during the last 16 h before calving. The average value at the time of calving was 19.2 ng/ml with a significant drop 1 h after calving to 11.2 ng/ml and a return to basal concentrations by 3 days postpartum. Evidence that labor is responsible for the increased corticoid is given. Maternal cortisol at calving was correlated with the time taken to calve .23, calf cortisol .04, cow:calf weight ratio -.07, and calf weight .14.


Fasting heat production in "lactating" versus dry dairy cows. Fasting heat production was measured 1 and 31 days after lactation ceased in postabsorptive dairy cows varying in daily rate of milk yield just prior to fasting. Daily fasting heat production was higher at 1 than at 31 days, particularly in higher-producing cows. It averaged 100 +/- 3 kilocalories per kilogram body weight-75 31 days after lactation ceased.


Selenium content of feeds and effect of dietary selenium on hair and blood serum. A selenium depletion diet (.025 ppm selenium) was fed to two male Holstein calves for its effect on concentrations of selenium in serum. By 90 days, selenium in serum was depressed from .022 ppm to .013 ppm, and one of two calves had died of "possible white muscle disease." Repletion was rapid in that a diet containing .200 ppm selenium increased selenium in serum to .028 ppm within 2 wk. Dietary .280 ppm selenium increased it to .051 ppm in 1 wk. Eleven cows and eleven calves showed averages of .030 ppm in serum (range of .013 to .051 ppm) fed a diet of corn silage (.039 ppm) and supplement (.090 ppm). A sampling of feedstuffs showed a wide range of selenium among feedstuffs (.023 for corn cobs to 2.663 ppm for dried egg albumin) and a wide range within one type feedstuff (shelled corn, .017 to .219 ppm).


Detection of allergy to nuts by the radioallergosorbent test. The diagnosis of food allergy is often difficult to make by conventional means. Histories are frequently ambiguous, and skin testing is of dubious reliability because of the number of false-positive and false-negative reactions. We have evaluated the radioallergosorbent test (RAST) for the in vitro measurement of the specific IgE antibodies to nuts, including Brazil nut, almond, walnut, pecan, cashew, and the legume, peanut. Serums were obtained from 18 patients with a history of nut allergy and IgE level and specific IgE antibodies were measured. Thirteen of the 18 patients had significantly elevated IgE antibody (greater than twice control) to one or more of the allergens. Prausnitz-Küstner tests on selected serums in general corroborated the results of the in vitro studies. Five patients had RAST elevations to 2 or more nuts. As a group RAST-positive patients had elevated mean serum IgE levels and more severe clinical symptoms (p less than 0.01). The specificity and cross-reactivity of IgE antibodies to different nut antigens was investigated by RAST inhibition with serums from 5 patients having high levels of IgE antibody. In 4 patients no cross-reactivity between Brazil nut and peanut was found. In contrast, several nut extracts inhibited the reaction of pecan allergen with IgE antibodies. These results indicate that specific IgE antibodies can be measured by RAST in patients with nut allergy and the cross-reactivity of nut antigens can be investigated. RAST would appear to be most useful in confirming the diagnosis of nut hypersensitivity in children or in highly allergic patients in whom skin testing poses a risk of anaphylaxis.


The occurrence of arrhythmias in hospitalized asthmatic patients. Twenty-four hour continuous ECG monitoring was performed on 20 patients hospitalized for asthma to evaluate the occurrence of potentially dangerous arrhythmias. Most patients (19/20) had sinus tachycardia on admission; premature ventricular contractions (PVCs) were present in 4; 1 had angina associated with runs of increased sinus tachycardia. With improvement in PaO2 and FEV1, the heart rate decreased in 17 patients and fewer PVCs were present in 2. Arrhythmias were not a serious problem in the patients studied.


Activation of the complement sequence by extracts of bacteria and fungi associated with hypersensitivity pneumonitis. Complement (C) consumption by antigens associated with hypersensitivity pneumonitis was investigated. Micropolyspora faeni in the presence of precipitating antibodies consumed C in a manner similar to the BSA-anti BSA system. An extract of M. faeni was shown to consume C and convert C3PA to C3A in the absence of detectable antibodies. In addition, three species of Aspergillus and two strains of Thermoactinomyces vulgaris were also shown to have the same activity with respect to C3PA. Mucor racemosus, Hormodendrum sp. and a mixed extract of Penicillium did not convert C3PA.


Median age at onset of asthma and allergic rhinitis in Tecumseh, Michigan. The ages at which allergies are likely to first become manifest is a subject of epidemiologic interest. The distribution of the onset ages reported in a population survey provides a distorted picture because it is a function of the age distribution of the population at the time of survey, and hence does not represent generational experience. A lifetable method is presented which transforms the raw cross-sectional data to a longitudinal basis. The information on onset age of asthma and allergic rhinitis collected in the 1962-1965 cycle of the Tecumseh Community Health Study is used to illustrate the method. The resultant distributions have median onset ages for the respective allergies which are considerably higher than those based on the onset frequency distributions of enumerated data.


Wood dust hypersensitivity.

Wood dust hypersensitivity. Two patients developed severe respiratory difficulty after occupational exposure to sawdust derived from abiruana wood. Both patients had positive immediate wheal-and-flare responses to skin tests with an extract of abiruana wood. Skin tests to other wood dusts were negative. Precipitating antibodies could not be demonstrated in the patients' serum. Bronchial challenge in both patients produced acute airway obstruction that was rapidly reversible. In addition, 1 patient developed airway obstruction 6-8 hr later that was more gradually reversible.


Egg allergy, influenza vaccine, and immunoglobulin E antibody. In a study of 70 patients with asthma, rhinitis, and eczema, those giving a definite history of allergic reactions to egg more frequently showed positive skin tests to egg extracts (p = less than 0.003), the wheal diameters of which were significantly larger (p = less than 0.01) than in patients with only a possible or no such history. Patients with a definite history of egg allergy had significantly higher levels of specific IgE antibody against egg yolk, egg white, and allantoic fluid than patients in the other two groups (p = less than 0.005). Seven patients, all of whom had given a definite history of allergy to egg, were found to have positive skin prick tests to influenza vaccine, at the concentration used in medical practice. Two of these patients had previously been given influenza vaccine and both had developed adverse reactions. Of the 22 patients giving a definite history of allergy to egg, the 7 (35 per cent) with positive skin tests to influenza vaccine had significantly larger skin tests and higher levels of specific IgE antibody to the egg extracts than the group as a whole (p = less than 0.001). Allergic reactions to influenza vaccine are likely to occur in patients who have a definite history of allergy to egg and large skin prick test reactions or high levels of specific IgE antibody to egg extracts. Those at risk can best be identified by skin prick testing with egg extracts and undiluted influenza vaccine.


Competent cellular immunity in allergic rhinitis patients with elevated IgE. Parameters of cell-mediated immunity (CMI) were studied in 17 allergic rhinitis patients selected for markedly elevated total serum IgE levels (greater than 300 IU/ml) and 14 normal controls. Mean serum IgE levels were 1,421 IU/ml and 101 IU/ml in the allergic and control groups, respectively (p less than 0.001). There were no significant differences between the allergic patients and the normal controls in delayed cutaneous hypersensitivity, in mitogen and antigen lymphocyte transformation in heterologous or autologous plasma, or in percentage of sheep erythrocyte rosettes. The allergic patient group had a significantly higher percentage of sheep erythrocyte-antibody-complement rosettes (p less than 0.05). Markedly elevated total serum IgE levels in allergic rhinitis patients were not associated with any detectable impairment of CMI.


Effect of treadmill exercise on asthmatic children. Measurement of peak expiratory flow rate before and after treadmill walking repeated at hourly intervals in asthmatic children disclosed progressive decreases in exercise-induced asthma. Treadmill running was followed by more extreme excercise-induced bronchospasm than treadmill walking. Exercise-induced asthma occurred in some children whose heart rates did not reach 160 during treadmill exercise, and heart rate during exercise was not correlated with the extent of exercise-induced bronchospasm.


Properties of mouse homocytotropic and heterocytotropic antibodies. Mouse antisera were analyzed for the presence of homocytotropic and heterocytotropic antibodies. Two distinct populations of antibodies were detected by the homologous passive cutaneous anaphylaxis (PCA) reaction. The first was active 2 hr after injection, heat-stable, and partially reactive with antisera to mouse IgG1. The second was active 48 hr after injection, was heat-labile, and probably belonged to the IgE class of mouse immunoglobulins. In addition, it was demonstrated that there were also two antibodies active in the heterologous PCA reaction in rats, a heat-stable antibody and a heat=labile antibody. Contrary to results obtained with homocytotropic antibodies, none of the heterocytotropic antibodies detected reacted with antisera to mouse IgG1 or IgG2. These studies suggest that in addition to IgE there may exist another heterocytotropic antibody in mouse antisera and that caution should be employed when using the 2-hr PCA reaction in the rat as a sole criterion for detection of mouse IgE.


Allergenicity and immunogenicity of Basidiomycetes. Species selected from six families of the class Basidiomycetes were evaluated for allergenicity in atopic and nonatopic individuals and for immunogenicity and antigenic cross-reactivity in experimental animals. Between 42% and 68% of atopic asthmatics demonstrated positive Type 1 wheal-and-flare skin reactivity to basidiomycete metabolic and somatic antigens. Sixty-four percent of skin test-positive atopic asthmatics exhibited positive RAST to as basidiomycete metabolic antigen and 50% were positive to somatic antigen. Negative RAST results were obtained in all nonatopic control sera. Only an occasional individual demonstrated positive serum antibasidiomycete precipitins by counterimmunoelectrophoresis. All basidiomycete species studied were highly immunogenic in the rabbit and most appeared to contain an electrophoretically heterogenous group of antigens with predominant anodal mobility. Ouchterlony double-diffusion analysis employing hyperimmune rabbit antisera indicated the presence of shared antigens among all basidiomycete species with the exception of Pleurotus. Rabbit antibasidiomycete sera did not cross-react with antigens of several common species of the Fungi imperfecti. Results indicate that many atopic asthmatics of the Gulf South area demonstrate IgE-medicated hypersensitivity to basidiomycete antigens as evidenced by positive wheal-and-flare skin reactivity and/or RAST. Basidiomycetes are also immunogenic in the rabbit and possess antigens that do not cross-react with those of certain Fungi imperfecti.


Possible asbestos hazards in clinical allergy. The potential hazards of biological products contaminated by asbestos fibers are quite properly being reviewed by the food and pharmaceutical industries. We have demonstrated that glycerinated saline solutions can be contaminated with asbestos fibers under certain conditions of simulated clinical use of asbestos-packed syringes. Because the fibrogenic and carcinogenic effects of asbestos are subtle and may not be manifest for a long latent period, the future use of asbestos containing materials in allergy practice might be more harmful than now realized.


Changing patterns of professional education in colleges and universities--1964 and 1974. Survey in the North Atlantic Region. This report concerns two surveys of undergraduate nutrition education in a selected group of degree-granting institutions within the North Atlantic Region of the United States. The purpose of the surveys was to determine the status of nutrition teaching in the early 1960s and the direction of change during the intervening ten-year period (1974). Reviews of the literature and college catalogues, personal interivews, letters of inquiry, and questionaires formed the basis for study. The last decade has witnessed change in organizational arrangement and in the number of programs offering major concentrations in foods, nutrition, and dietetics. Of these, twelve units in the Region studied are now engaged in coordinated programming. There has been a 125 per cent increase in foods and nutrition faculty. All but seventeen of the 275 faculty appear in programs approved by The American Dietetic Association. Basic questions about undergraduate education in foods and nutrition and the most desirable administrative structure for this specialization will continue through the 1970s. Solutions will undoubtedly be forthcoming after further experimentation and evaluative research.


Development of a rating scale to measure learning in clinical dietetics. I. Theoretical considerations and method of construction. The trend to introduce earlier clinical experience in dietetic education requires that attention be directed toward evaluation of learning of such clinical skills as communication and interpersonal relations. Some of the theoretical factors associated with clinical evaluation are discussed, and a method is described for developing a rating scale to measure learning in an intoductory clinical course in dietetics. The method involved: (a) analysis of course objectives to determine the general areas of learning which were appropriate and valid to measure in the clinical environment; (b) specification of observable and one-dimensional behaviors seen as componets of the overall objectives; and (c) concise definition of the rating continuum. Specific attention was directed toward constructing an instrument which could be used to counsel students as well as provide information for determining grades. Preliminary trials indicated the instrument to be practical, reliable, objective, and valid to measure clinical learning.


Development of a rating scale to measure learning in clinical dietetics. II. Pilot test. When the instrument was pilot tested under actual conditions of a clinical course, it was found practical to use, provided the raters were sufficiently trained and were aware of the logistical problems that might occur. A consensus of both students and raters reaffirmed that the formative properties of the instrument were of value in student counseling, and the rating of students during each clinical session should continue. The objectivity of the instrument also appeared to be sufficient to allow summative grading of students without large differences of opinion within pairs of raters. Measurement of intra-rater reliability was not attempted. The investigators feel that any future study with the instrument should include this analysis. The process of construction and testing reported here appears to be useful in developing a rating scale to measure learning in clinical dietetics.


Design and use of an instrument to evaluate students' clinical performance. An instrument for evaluating clinical performance of students was developed for the coordinated undergraduate program in dietetics at The Ohio State University. The development and evaluation of the instrument are described. Statistical data show a significant degree of correlation among three evaluators: the student, his preceptor, and faculty. In addition, use of the instrument facilitated evaluation, and students responded favorably to criterion-referenced rather than to norm-referenced evaluation conferences.


Nutritional knowledge and attitudes of registered nurses. Nutritional knowledge and attitudes of hospital nurses in Nebraska and their perceptions of the role of the dietitian in patient care were studied. In general, they had favorable attitudes toward nutrition. Although their knowledge was adequate, they lacked confidence in that knowledge. The nurses perceived dietitians in an educational rather than a service role. Older nurses had more favorable attitudes but were less knowledgeable than younger ones. More knowledgeable nurses had more positive attitudes toward their own role in nutrition education and toward the team approach to health care.


Developing a demand forecasting system for a foodservice operation. In foodservice operations, accurate and dependable forecasts of food production demands can help control food and labor costs. A decreased incidence of menu item over- and under-production should lower scheduled labor and production time and optimize use of equipment. Each foodservice system has specified characteristics and patterns of activity. A procedure to develop, establish, control, and evaluate a forecasting system is described. The objectives of the foodservice and the proposed forecasting system must be defined. A cycle menu and historical data bases are two key inputs. It is more accurate to forecast menu item demand than diet category demand because of the complexity in categorizing multi-restricted diets. Control of the system is maintained by establishing policies and procedures and conducting routine subjective and objective evaluations.


A weight control program for students using diet and behavior therapy. A twelve-week weight-reducing program for students was conducted at the University of Kentucky. The program, which was developed and directed by graduate students in nutrition, utilized a combination of behavior therapy and diet therapy. Initiated in the spring of 1975, the program has been continued in the 1975-76 year. All students completing the program achieved weight reduction. Their weight losses averaged 2/3 lb. a week. The dropout rate for the program was 40 per cent. One-third of the participants completing the program reached their weight-reduction goals. These results suggest that further attempts should be made to improve weight-reducing suitable for college students.


Progeria: autopsy report of one case, with a review of pathologic findings reported in the literature. An autopsy case of progeria associated with elastotic degeneration of the skin and preservation of the scalp hair, in a 20-year-old woman, is presented. The pathologic findings were scleroderma-like skin atrophy, elastotic degeneration of the skin, arteriosclerosis, atrophy of the endocrine glands, and acute peritonitis after appendectomy. In the reported cases of progeria, as in Werner's syndrome, no responsible parenchymatous organ has been identified. Rather, connective tissue, widely distributed throughout the entire body, may play  an important role. The possible abnormal metabolism of connective tissue in progeria deserves further study.


Free radical theory of aging: inhibition of amyloidosis in mice by antioxidants; possible mechanism. The antioxidants, alpha-tocopherol acetate and a quinolone derivative (Santoquin), inhibited the development of amyloidosis when added to the diet of casein-injected C3HeB/FeJ male mice. Santoquin, and to a lesser extent butylated hydroxytoluene (BHT), also depressed the appearance of a plasma protein fraction in these mice; the effect of alpha-tocopherol was not determined. Consideration of the current knowledge of amyloid, in the light of these antioxidant studies, prompted the following hypothesis. Amyloidosis is largely the result of an enhanced rate of oxidative degradation of a connective-tissue glycoprotein(s) coupled with oxidative/enzymatic changes in the plasma, both of the tissue-derived substances and of immunoglobulins, to form the amyloid fibril protein subunits (AL and AA) which subsequently aggreagate to form the amyloid fibrils.


Comparison of two systems for stroke rehabilitation in a general hospital. This study was designed to compare the effectiveness of stroke rehabilitation therapy in a specialized Stroke Unit with that provided on the medical service of a general hospital (Rhode Island Hospital). The 8-bed Stroke Unit is staffed by a multidisciplinary team, and a weekly conference is held for evaluation and planning. On the basis of data obtained from the hospital records, two groups of patients were studied: 224 who were treated in the Stroke Unit, and 110 who were evaluated and approved for admission to the Unit but were not accommodated. A rigid "first come, first served" policy for admission to the Unit was observed. Hypothesis testing was performed with reference to the patient's medical condition, socioeconomic status, demographic characteristics, and difficulties during hospital stay to determine whether the groups were comparable. A patient was considered to have improved if his condition decreased in severity between the time of admission to therapy and the time of discharge. Severity was rated as: mild (level one), moderate (level two), severe (level three), and profound (level four). No significant difference in rehabilitation results was found between the two treatment systems at severity levels two (moderate) and four (profound). However, the Stroke Unit attained significantly better results with level-three patients (severe stroke). This group received more sessions of physical therapy and remained in the hospital longer than did the level-three patients treated on the general medical service. Physicians referred patients selectively to the Stroke Unit, althoug the Unit had no policy of screening patients for admission, and this may have had some influence on the achievement of better results with level-three patients. Level-four patients did not do well in either setting.


The care of decubitus ulcers pressure sores. Despite a large volume of literature particularly directed toward treatment, pressure sores (including decubitus ulcers) remain a difficult problem, especially in the nursing home environment. The treatment of pressure sores is somewhat controversial and quite diversified. Selection of a successful therapeutic modality must be preceded by correct evaluation, i.e., whether the sore is superficial or deep, open or closed. The treatment of superficial sores is conservative and directed toward cleanliness, relief of pressure, and exposure to air. Surgical debridement may be indicated. Proteolytic enzymes often are employed as adjunctive therapy, although there are some major drawbacks to their use. The plethora of therapeutic agents suggested for the treatment of deep pressure sores probably is related to the difficulties in achieving success. Surgical debridement is indicated, and proteolytic enzymes are widely used. Possible interactions. and factors leading to the inactivation of these enzymes are discussed, as is the use of various solutions, ointments, gold leaf, oxygen, dry heat, and other adjunctive devices. Of paramount importance in the management of pressure sores is the maintenance of cleanliness and dryness.


Myasthenia gravis in the elderly.

Myasthenia gravis in the elderly. During a 15-year period in Jerusalem, myasthenia gravis was diagnosed in 15 patients aged 60 or older. At this age, diagnosis is difficult because of such possible associated conditions as cerebrovascular and cardiovascular arteriosclerotic diseases or bronchopulmonary disorders. The clinical course often is complicated by these associated conditions, and also by their treatment. Seven of the patients died within 4 years of the onset of their illness, but only 5 because of myasthenia gravis. This disease represents an important diagnostic and therapeutic problem in elderly patients.


Immunisation of sheep against Schistosoma mattheei using either irradiated cercariae or irradiated schistosomula. Irradiated cercariae, irradiated schistosomula, or heterologous infections were used to vaccinate sheep against Schistosoma mattheei infection. In the first experiment four doses of 10(4) S. mattheei cercariae irradiated at 6Kr were administered to sheep by percutaneous infection at 4 week intervals. This induced a 74% reduction in a challenge infection compared to control sheep while only 13% protection was achieved in a third group of sheep immunised with normal cercariae of the heterologous parasite S. mansoni. No significant differences were seen in histopathology of the liver of any of the sheep but the pathological changes were more severe in the large and small intestines of sheep vaccinated with the heterologous parasite. In the second experiment with irradiated cercariae only one or two immunising exposures were used. The degree of protection in the adult worm load (9-11%) was not significant and no significant differences were noticed in the pathology of the vaccinated and control animals. In the third experiment four doses of irradiated organisms were used to vaccinate five groups of sheep: 3Kr or 6Kr cercariae were administered by percutaneous infection; 6Kr skin-transformed schistosomula were administered by intra muscular injection; the same 6Kr skin-transformed schistosomula were given by intravenous injection and 6Kr syringe transformed schistosomula were administered by intramuscular injection. The degree of protection (determined as the reduction in worm burden) achieved by these different procedures was respectively 72%, 61%, 77%, 56% and 78%. These results indicate the possibility of making a live vaccine against ovine schistosomiasis and show that effective immunisation is not dependent on the presence of a mature worm infection or on cercarial penetration of the skin by the immunising infection.


Comparative histochemical observations on the lipids in the immature and mature stages of Cotylophoron cotylophorum Paramphistomatidae: Digenea. Immature and mature stages of the sheep amphistome Cotylophoron cotylophorum have been analysed histochemically for their lipids. Excretory ducts of the immature worms were the common sites for the accumulation of neutral lipids (triglycerides) and phospholipids which showed very sparse distribution at the corresponding sites in the adult. Phospholipids and lipoproteins observed in the tegument of the adult could not be seen in the immature form. Intestinal caecae of both forms showed the presence of diffused and granular lipids which were relatively less in the immature. The possible physiological significance of these differences in the lipid contents of the immature and mature forms has been discussed.


Comparison of the Bell technique, a modified Kato thick smear technique, and a digestion method for the field diagnosis of schistosomiasis mansoni. A modification of the Kato thick smear technique, the Bell filtration technique, and a digestion method were investigated to determine the relative merits of each for the detection of schistosome eggs in human faecal samples. The modified Kato technique was the method of choice because it was sensitive, simple to perform, and suitable for field use. The Bell filtration technique was equally sensitive in detecting light infections, but was more tedious and more complicated to perform than the thick smear technique.


Studies with Brugia pahangi 12. The activity of levamisole. The effects of levamisole on adults, third stage infective larvae, and microfilariae of Brugia pahangi were studied in in vitro culture and in vivo against developing stages in the vector mosquito and in infected cats. In vitro the drug was effective only at dose levels much higher than can be tolerated by mammals. It was active against the developmental stages of the worm in the vector Aedes aegypti. The drug was strongly microfilaricidal in cats but less effective against adult worms.


Echinobothrium harfordi sp. nov. (Cestoda: Diphyllidae) from Raja naevus in the North Sea and English Channel. Echinobothrium harfordi sp. nov. from the spiral intestine of Raja naevus in British waters is described and illustrated. The species most closely resembles E. affine as both possess two groups of eleven large apical hooks, but can be distinguished by fewer peduncle spines and by characteristics of the eggs. The intensity of infection in individual hosts was generally low, and although the incidence of infection was high in rays less than 35 cm in length, this fell towards zero in larger rays. The cestode showed a marked affinity for the first two tiers of the spiral intestine.


A comparative study of the death of schistosomula of Schistosoma haematobium and Schistosoma mansoni in the skin of mice and hamsters. This study shows that some cercariae of S. haematobium and S. mansoni die during penetration of mouse or hamster skin. Approximately 30-38% of cercariae of both species die in mouse skin and 14-16% die in hamster skin. The greater number of cercariae which die in the skin of mice seems to account for the higher yield of adult worms recovered in hamsters. Adult worm recoveries from animals infected with S. haematobium were, however, only about half the worm recoveries from hosts infected with S. mansoni.


Intermediate host antigens associated with the cercariae of Schistosoma haematobium. Bulinus (Physopsis) africanus antigens were shown to be associated with the cercarial glycocalyx of Schistosoma haematobium using immunofluorescence and the Cercarienhüllen Reaktion. It is proposed that this snail antigen may sensitise the definitive host and that resistance to further invasion by cercariae could be induced in this manner.


The anthelmintic effects of a new compound "E" (Friedheim) on Onchocerca gutturosa in the cow--a possible tertiary screening system for drug action against O. volvulus in man. Three cows were treated with different levels of compound "E" which is known to have strong filaricidal activity. In all three animals the number of microfilariae of Onchocerca gutturosa declined after treatment. At autopsy most of the adult worms found were dead and in those still living embryogenesis appeared to have ceased. It is concluded that, despite the disadvantages, this system represents a possible way of assessing onchocercicidal activity before undertaking human trials.


The detection of antibodies in human and animal filariases by counterimmunoelectrophoresis with Dirofilaria immitis antigens. Counterimmunoelectrophoresis revealed the presence of precipitin antibody in all of 6 dogs and the 1 cat infected with Dirofilaria immitis and in the serum of 17 of 24 individuals living in a setting of hyperendemic subperiodic bancroftian filariasis. Antigens used in the test were prepared from microfilariae and adult male D. immitis. Some humans and animals had antibodies to both antigens while others had antibodies against microfilariae or adult worms only. The presence of soluble circulating antigen was detected in the sera of two dogs with high microfilaraemias.


Ultrastructural cytochemistry of p-N,N-dimethylamino-beta-phenethylamine (DAPA) oxidation reactions. The ultracytochemical localization of amine oxidase (AO) activity is demonstrated with a new substrate, p-N,N-dimethylamino-beta-phenethylamine (DAPA). DAPA was designed to yield a stronger reducing agent on oxidation by monoamine oxidase (MAO) than is obtained from the MAO substrate, tryptamine, upon oxidation. Thus MAO and possibly other oxidase(s) can be demonstrated with DAPA and the tetrazolium salt, 2-(2'-benzothiazolyl)-5-styryl-3-(4'-phthalhydrazidyl) tetrazolium chloride (BSPT). The latter is a nonosmiophilic tetrazolium salt which is reduced to an osmiophilic formazan. In addition, DAPA itself demonstrates AO activity ultracytochemically with and without BSPT. We speculate that either oxidative polymerization of DAPA or Schiff's base formation with protein after aldehyde formation is responsible for the latter reaction, which is made permanent for ultracytochemical localization by osmication at a later step. DAPA oxidation reaction products are demonstrated in guinea pig kidney, specifically in the endoplasmic reticulum, nuclear envelope and mitochondrial outer compartments and cristae. Differences in reaction product characteristics and localization in relation to formaldehyde fixation and the localization of reaction product in mitochondrial cristae, as well as outer compartments, suggest that DAPA oxidation is mediated through one or more MAOs and possible other oxidases.