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Biomedical subjects

W Ma

Publications and source records attributed to W Ma.

At least 19 recordsLinked to original sources

Inhibitory effect of green tea on the growth of established skin papillomas in mice.

In 10 separate experiments, mice with established chemically induced or UV light-induced skin papillomas were treated continuously with green tea in the drinking water or with i.p. injections of a green tea polyphenol fraction or (-)-epigallocatechin gallate three times a week for 4-10 weeks. Partial tumor regression or > 90% inhibition of tumor growth, as measured by changes in tumor volume per mouse, was observed in 5 experiments, and marked inhibition of tumor growth (46-89%) was observed in 5 additional experiments. Treatment of the mice with green tea or green tea constituents had an inhibitory effect on body weight increases in several but not all of the studies. Examination of the data from all ten experiments revealed that complete tumor regression occurred in 14 of 346 papilloma-bearing mice (4%) that were treated with green tea in the drinking water or with i.p. injections of green tea constituents, whereas none of the 220 papilloma-bearing control mice treated with only vehicle exhibited complete tumor regression. These observations indicate that oral administration of green tea, i.p. administration of a green tea polyphenol fraction, or i.p. administration of (-)-epigallocatechin gallate inhibited the growth and/or caused the regression of established experimentally induced skin papillomas.

9,10-Dimethyl-1,2-benzanthracene

Neuroepithelial cells in the rat spinal cord express glutamate decarboxylase immunoreactivity in vivo and in vitro.

It is unknown whether neuroepithelial cells in the mammalian central nervous system express neurotransmitter-synthesizing enzymes. In this study, expression of glutamate decarboxylase (GAD), the gamma-aminobutyric acid (GABA)-synthesizing enzyme, was examined in proliferative cells and postmitotic neuroblasts in embryonic rat spinal cord. Immunostaining coronal sections of the embryonic spinal cord with K2 antiserum, which recognizes GAD proteins encoded by the GAD67 gene, revealed intensely stained neuroepithelial cells in the basal plate at embryonic day (E) 13, in the intermediate plate between E 13-16, and last seen in the alar plate at E 16. Nissl counterstaining demonstrated that a small number of these GAD-immunoreactive cells adjacent to the neural tube lumen were mitotic. The ventral-to-dorsal gradient of GAD expression in precursor cells and postmitotic neuroblasts correlates anatomically and temporally with the sequential generation of motoneurons, commissural neurons, and interneurons in the dorsal horn. Some of these GAD-immunoreactive neuroepithelial cells may re-enter the mitotic cycle, while others are postmitotic neuroblasts presumably migrating to the intermediate zone to differentiate into young neurons. Double-immunostaining cells acutely dissociated from E 11-18 spinal cords with K2 and anti-bromodeoxyuridine antisera, following a bromodeoxyuridine pulse in vivo, revealed considerable numbers of DNA-synthesizing cells immunoreactive for GAD. The absolute number of double-stained cells peaked during E 12-15, coinciding with terminal cell division in most spinal neurons. These observations suggest that spinal neuronal precursors can synthesize GAD-related proteins prior to, or during, the terminal cell cycle. Although GAD immunoreactivity revealed by K2 antiserum was detected in proliferative cells and in migrating postmitotic neuroblasts, GABA immunoreactivity was never detectable in these cells. These early embryonic GAD-immunoreactive neuroepithelial cells may either synthesize levels of GABA that cannot be detected immunocytochemically, and/or express enzymatically inactive GAD-related proteins.

Animals

Transient expression of GABA immunoreactivity in the developing rat spinal cord.

The development of GABAergic neurons in the spinal cord of the rat has been investigated by immunocytochemical staining of frozen sections with anti-gamma-aminobutyric acid (GABA) antiserum. In the cervical cord, GABA-immunoreactive fibers first appeared at embryonic day (E) 13 in the presumptive white matter within the ventral commissure, ventral funiculus, and dorsal root entrance zone, and in the ventral roots. There were no GABA-immunoreactive cell bodies detected at this age. By E14, motoneurons, the earliest generated spinal cells, were the first cell population to become GABA-immunoreactive at the cell body level. Thereafter, GABA-immunoreactive neurons increased progressively in number and extended from ventral to dorsal regions. GABA-immunoreactive relay neurons within lamina I of the dorsal horn were initially detected at E17. Interneurons in the substantia gelatinosa, the latest generated cells in the spinal cord, were also the last to express the GABA immunoreactivity at E18. Immunoreactive neurons peaked in intensity and extent at E18 and 19. GABA immunoreactivity was only detectable in neurons within the intermediate and marginal zones 1-3 days after they withdrew from the cell cycle. This contrasts to glutamate decarboxylase immunoreactivity, which is detected in precursor cells in the ventricular zone prior to, or during, withdrawal from the cell cycle. Toward the end of gestation, GABA immunoreactivity declined in intensity and extent. This regression began in the ventral horn of the cervical region and ended in the dorsal horn of the lumbosacral region. During the first week after birth, immunoreactivity in motoneurons and in many other neurons within the ventral horn, intermediate gray, and deeper layers of the dorsal horn disappeared, and only in those neurons predominantly within the superficial layers of the dorsal horn did it persist into adulthood. Thus, the expression and regression of GABA immunoreactivity in the spinal cord followed ventral-to-dorsal, rostral-to-caudal, and medial-to-lateral gradients. These observations indicate that the majority of embryonic spinal neurons pass through a stage of transient expression of GABA immunoreactivity. The functional significance of this transient expression is unknown, but it coincides with the period of intense neurite growth of motoneurons, sensory neurons, and interneurons, and of neuromuscular junction formation, suggesting that the transient presence of GABA may play an important role in the differentiation of sensorimotor neuronal circuits.

Animals

Cholinergic depletion prevents expansion of topographic maps in somatosensory cortex.

Although the role of acetylcholine in processing stimuli in the cerebral cortex is becoming defined, the impact of cholinergic activity on the character of cortical maps remains unclear. In the somatosensory cortex, topographic maps appear capable of lifelong modifications in response to alterations in the periphery. One factor proposed to influence this adaptational ability is the presence of acetylcholine in the cortex. The studies presented here, using the 2-deoxyglucose technique, demonstrate that the unilateral removal of a digit in cats, followed by stimulation of an adjacent digit, produces a pattern of metabolic activity in the somatosensory cortex that is dramatically expanded when compared with the opposite (normal) hemisphere. In contrast, experiments in which the somatosensory cortex was depleted of acetylcholine and the animal received a similar amputation led not to patterns of expanded metabolic activity, but rather to reductions in the evoked metabolic distribution. These studies implicate acetylcholine in normal map formation and in the maintenance of the capacity of cortical maps to adapt to changes in the periphery.

Acetylcholine

The relationship between thalamocortical connections and stimulus-evoked metabolic activity in the ventroposterior nucleus of the monkey.

Although a highly organized system of reciprocal projections exists between the cerebral cortex and the thalamus, the relationship of the thalamocortical projections to functional activity remains unclear. This study attempts to identify the correlation between thalamic relay cells and functional activity evoked in the ventroposterior nucleus (VP) of cynomolgus and squirrel monkeys. Wheatgerm agglutinin conjugated to horseradish peroxidase (WGA:HRP) was iontophoretically injected into physiologically determined sites in the somatosensory cortex, resulting in retrogradely labeled cells and anterogradely labeled terminals in corresponding somatosensory thalamic regions. In the same animals, 2-deoxyglucose (2DG) experiments were carried out 2 days later, using the somatic stimuli identified as best exciting the cortical neurons. Stimulation to the limbs produced crescent-shaped clusters of metabolic label arranged in a somatotopically organized fashion in the ventral posterior lateral nucleus (VPL). Following WGA:HRP injections into area 3b, the stimulus-evoked 2DG label was colocalized with the retrograde and anterograde tracer. This finding suggests that the location of stimulus-evoked metabolic activity can be predicted by the presence of transported WGA:HRP clusters.

Afferent Pathways

Cholinergic manipulation alters stimulus-evoked metabolic activity in cat somatosensory cortex.

The role of acetylcholine (ACh) in cerebral cortical activity has recently been reevaluated. It now seems clear that this neurotransmitter increases the magnitude of cortical responses. Although substantial information has been gathered regarding the role of ACh in sensory information processing, little is known about the participation of ACh in the organization of maps in the cerebral cortex. To address this issue, we used 2 methods to manipulate the supply of ACh in the somatosensory cortex of cats: 1) unilateral neurotoxic lesions of the basal forebrain and 2) unilateral topical applications of the cholinergic antagonist, atropine. For each experimental condition, the animal received an injection of 2-deoxyglucose (2DG) while identical somatic stimuli were delivered to the right and left forepaws. In the somatosensory cortex, the 2DG uptake most often occurred in the form of patches that extended from layer II to IV. When the patches were reconstructed into 2-dimensional maps of activity throughout the somatosensory cortex, they formed strips that ran in the rostrocaudal direction. The reconstructed maps revealed that the 2DG patterns in ACh-depleted and the normal cortex were similar in their overall topographic distribution. Depletion or antagonism of ACh, however, caused the stimulus-evoked metabolic label to be reduced in dimension and density. Measurements of background activity levels were obtained by using 1) cytochrome oxidase histochemistry or 2) metabolic activity values in regions of somatosensory cortex that were not specifically stimulated. This analysis indicated that background values in the ACh-depleted hemispheres were not different from those in the normal hemispheres. The absence of ACh therefore appears to reduce the cortical response to stimulation, while background activity values do not change. These observations indicate that ACh plays a significant role in the processing of sensory information and the organization of somatosensory cortical maps.

Acetylcholine

[Detection of antibodies to citric acid extract of skeletal muscle (CAE-ab) in the sera of patients with myasthenia gravis using indirect hemagglutination].

In this paper are described the method and results for the demonstration of the antibodies to citric acid extract of skeletal muscle (CAE-ab) in the sera from myasthenia gravis (MG) patients with or without thymoma, in patients with other diseases and in healthy controls by an indirect hemagglutination assay (IHA), were described. The CAE-ab titers were positive in 15 (71.43%) of the 21 MG patients with thymoma, the antibody titers ranging from 1:16 to 1:512. In the 35 MG patients without thymoma, as well as in the 32 healthy controls and 22 patients with other diseases, the antibody titers were all negative A statistically significant difference was found between the CAE-ab titers in the group of MG patients with thymoma and those in the other three groups. It was considered that IHA for the purpose of demonstrating CAE-ab could serve as a supplementary diagnostic method for early detection of thymoma in MG patients at an early stage.

Adolescent

Lesions of the basal forebrain alter stimulus-evoked metabolic activity in mouse somatosensory cortex.

The role that acetylcholine plays in processing sensory stimuli is beginning to be characterized; however, morphological correlates of cholinergic effects on activity patterns in sensory cortex are not available. To study this problem, unilateral neurotoxic lesions that depleted the necortex of acetylcholine were made in the basal forebrains of mice. The aim of these experiments was to study the effect of cholinergic depletion on stimulus-evoked activity in the barrel field of the mouse somatosensory cortex. One month following the lesion, 2-deoxyglucose (2DG) experiments were conducted on the lesioned and on normal mice while the animal received bilateral stimulation to the C3 whisker. The tissue was processed for acetylcholinesterase and cytochrome oxidase histochemistry and 2DG autoradiography. Evaluation of the column-like 2DG label evoked in the somatosensory cortex revealed that the activity on the lesioned side was significantly reduced in dimension and intensity from that in the normal hemisphere. On the normal side, the activated barrels averaged 641 microns in tangential width, were 76.5% above background in density, and extended from lamina I-V. On the lesioned side, the activated barrels were 485 microns in tangential width, 65.4% above background in density, and extended from lamina II-V. In other cortical regions, outside the stimulus-evoked barrel field, 2DG activity values were similar on the normal and lesioned side. Additionally, both the pattern and intensity of the cytochrome oxidase staining within the barrel field displayed no differences in either hemisphere. These studies suggest that acetylcholine plays a significant role in the processing of somatosensory information. Neurotoxic lesions that diminish cortical cholinergic innervation cause a reduction of stimulus-evoked activity levels, while underlying metabolic activity is either not affected or recovers over time.

Acetylcholine

Spinal input to the parabrachial nucleus in the cat.

The projections from the spinal cord to the parabrachial nucleus in the cat were investigated using both the degeneration method and the anterograde transport of wheat germ agglutinin-horseradish peroxidase conjugate. Both methods produced similar results. Spinal input to the parabrachial nucleus was bilateral, with a slight contralateral predominance. The termination area was localized predominantly in the dorsal part of the lateral parabrachial nucleus, with additional limited terminations in the Kölliker-Fuse subnucleus. Projections from different rostrocaudal levels of the spinal cord overlapped completely, suggesting that spinal input to the parabrachial nucleus is not topographically organized. Taking these results together with those of others indicating that spinal input to the parabrachial nucleus arises primarily from nociceptive-specific neurons in lamina I of the dorsal horn, it is concluded that the spinal projections to the parabrachial nucleus are likely to be involved in various generalized aspects of nociception.

Animals

Spino-diencephalic relays through the parabrachial nucleus in the cat.

Previous studies have shown that the spinal input to the parabrachial nucleus (PBN) in the cat is limited to certain portions of its lateral division 8,21,45. The purpose of the present study was to determine some of the output targets of PBN neurons located within this spinal terminal domain by means of single, double and triple light microscopic labeling strategies. Combinations of tracers included the retrograde transport of tritiated wheat germ agglutinin, wheat germ agglutinin conjugated with horseradish peroxidase (WGA-HRP) and Fluoro-Gold from the hypothalamus, amygdala or thalamus/zona incerta together with either the anterograde transport of WGA-HRP from the spinal cord or the degeneration of spinal terminals following spinal lesions. The results (summarized in Fig. 10) showed that the spinal terminal domain contains separable populations of neurons projecting to the thalamus/zona incerta and hypothalamus. Only a limited number of amygdala-projecting neurons was located in this domain. Evidence from several laboratories supports the conclusion that these potential spino-diencephalic relays are involved somehow in nociception. More information is needed, however, regarding differences in the response properties of these separable populations of spinal-recipient neurons before more specific hypotheses concerning the precise nature of their nociceptive functions can be formulated.

Animals

An ultrastructural analysis of serotoninergic neurons in the nucleus raphe magnus of the rat.

The synaptic organization of serotonin elements in the nucleus raphe magnus of adult rat was investigated by electron microscopic immunocytochemistry with an antiserum against serotonin itself. Immunoreactive somata in the nucleus raphe magnus were usually of the same size (7-15 microns) and showed similar cytological features as their unlabeled congeners. The serotonin perikarya were contacted by a few unlabeled axon terminals containing round synaptic vesicles, and gave rise to dendrites which often ran perpendicularly to the midline. These dendrites received many contacts from axon terminals containing round or pleomorphic synaptic vesicles, but themselves failed to show vesicular aggregates or membrane differentiations suggestive of synaptic specialization. Immunoreactive axon terminals were numerous and mostly contained round or pleomorphic vesicles. Several exhibited synaptic contacts on dendrites, but there were also others which did not show any synaptic membranous differentiation, even when followed in serial sections. Immunostained myelinated as well as unmyelinated axons could be observed. These results provide a first description of the morphology and synaptic organization of the serotonin neurons in adult rat nucleus raphe magnus.

Animals

Fine structure of the dorsal part of the nucleus submedius of the rat thalamus: an anatomical study with reference to possible pain pathways.

The dorsal portion of the nucleus submedius of the rat thalamus receives spinal and trigeminal projections which may convey noxious inputs. The present study was undertaken to analyse the fine structure of the nucleus with particular reference to a possible trigemino-thalamo-prefrontal cortical pathway relaying in nucleus submedius. Presynaptic terminals in the dorsal portion of the nucleus submedius were classified into three broad categories usually observed in thalamic nuclei: "small round", "flat" and "large round" types. Axonal tracing using either anterograde transport of horseradish peroxidase or degeneration methods indicated that some "small round" terminals originate from the pre-frontal cortex. Some "large round" terminals were labelled from the trigeminal subnucleus caudalis. These "large round" terminals exhibited distinct morphological features when compared with trigeminal terminals in other thalamic nuclei. In particular they made synaptic contacts predominantly with dendritic protrusions and were surrounded by multilamellate astrocytic processes. Double-labelling experiments were performed by means of the combined retrograde transport of horseradish peroxidase and Wallerian degradation techniques. Terminals degrading after lesion of the trigeminal subnucleus caudalis contacted submedius neurons labelled retrogradely from the prefrontal cortex. These observations demonstrate the existence of a direct monosynaptically relayed pathway from subnucleus caudalis to prefrontal cortex which relays in the dorsal part of nucleus submedius.

Afferent Pathways

Spinal and trigeminal projections to the parabrachial nucleus in the rat: electron-microscopic evidence of a spino-ponto-amygdalian somatosensory pathway.

The fine structure of spinal and trigeminal projections to the parabrachial area (PB) of the rat was studied using either the anterograde transport of a lectin-peroxidase conjugate or the degeneration technique. Two morphologically different types of terminals were observed. Most labeled terminals contained round vesicles (R type) and formed asymmetrical synapses, usually with large dendrites. Others contained pleomorphic vesicles (P type) and usually made symmetrical contacts with large or medium-size dendrites. A double-labeling strategy was used, combining the retrograde labeling of PB neurons with lectin-peroxidase conjugate from the amygdala and the identification of degenerating terminals after lesions of spinal or trigeminal pathways. These experiments demonstrated that spinal and trigeminal terminals contact PB neurons that project to the central nucleus of the amygdala. The role of this spino(trigemino)-ponto-amygdalian pathway is discussed in relation to some aspects of pain.

Afferent Pathways

Synaptic glomeruli in the nucleus submedius of the rat thalamus.

Synaptic glomeruli in the nucleus submedius of the rat are described and the source of some of the component terminals identified. The glomeruli consist of large terminals with round synaptic vesicles establishing Gray type I contacts with dendrites and surrounded by layers of astrocyte derived membranes. The astrocyte processes may be composed of cell membranes with minimal interventing cytoplasm or, less frequently, contain larger amounts of cytoplasm. Horseradish peroxidase injected into the trigeminal nucleus caudalis labels some of the large astrocyte-enclosed terminals in nucleus submedius.

Animals

Fine structure of the spinothalamic projections to the central lateral nucleus of the rat thalamus.

The fine structure of labelled spinothalamic terminals in the central lateral nucleus has been studied in the rat following injection of wheat germ agglutinin-horseradish peroxidase into the spinal cord. Myelinated axons gave rise to the labelled terminals, which were large profiles which contained round vesicles, numerous mitochondria, and formed asymmetrical contacts with large dendrites or dendritic protrusions. These profiles are similar to those described in other somatosensory thalamic nuclei, and in many other nuclei of the thalamus.

Animals

Swallow syncope after aneurysmectomy of the thoracic aorta.

A case of swallow syncope is presented and 30 previously reported cases from the literature in English are reviewed. A 67-year-old male developed syncope on swallowing 3 months after the resection of an aneurysm of the descending thoracic aorta. Electrocardiographic monitoring during eating demonstrated sinus bradycardia and sinus arrest with loss of consciousness. Neither Valsalva's maneuver nor carotid massage could produce bradycardia or syncope. Intravenous administration of edrophonium produced sinus bradycardia and the drinking of water by the patient after edrophonium administration brought about sinus bradycardia and sinus arrest with loss of consciousness. Sinoatrial node and atrioventricular node functions, evaluated by an atropine test, overdrive suppression test, and His bundle electrocardiogram were normal. No pathological changes were observed following a barium swallow. The patient was treated with a permanent pacemaker.

Aged

The differential synaptic organization of the spinal and lemniscal projections to the ventrobasal complex of the rat thalamus. Evidence for convergence of the two systems upon single thalamic neurons.

The synaptic organization of terminals originating either from the spinal cord (spinothalamic) or from the dorsal column nuclei (lemniscal) was investigated in the ventrobasal complex of the rat thalamus. Wheatgerm agglutinin conjugated to horseradish peroxidase was used as an anterogradely transported axonal tracer, using benzidine dihydrochloride as a chromogen for the identification by electron microscopy of spinal and lemniscal projections to the ventrobasal thalamus. A double anterograde tract tracing strategy, based labeling by wheatgerm agglutinin conjugated to horseradish peroxidase of spinal terminals and simultaneous visualization of lemniscal terminals identified by Wallerian degeneration induced by lesion of the neurons of origin in the dorsal column nuclei, was used to compare the postsynaptic elements contacted by the two pathways and to look for a possible convergence of the two pathways onto single thalamic neurons. Spinal and lemniscal terminals are large (2-2.5 microns mean average diameter) terminals containing several mitochondria and numerous rounded vesicles. A quantitative analysis of the mean average diameters of the terminals revealed that one could not differentiate between synapses formed by the two pathways on a morphological basis. Terminals of the two pathways make asymmetrical contacts (Gray type I) with dendrites of varying diameter, dendritic protrusions, and cell somata. A quantitative analysis of the least diameter of the postsynaptic elements demonstrates projections of the two systems to different, partially overlapping regions of thalamic neurons. Lemniscal terminals originating from the dorsal column nuclei frequently contact cell somata; axosomatic spinothalamic contacts are uncommon. In addition, lemniscal projections tend to contact more proximal dendrites than do spinal projections, and this differential synaptic organization is statistically significant. From a functional point of view, this differential synaptic organization might indicate that lemniscal inputs have greater influence than spinal inputs in affecting the activity of thalamic neurons. Labeled spinothalamic terminals contact the same dendritic profile as do degenerating lemniscal terminals in about 10% of single sections. Because the present study did not include a complete reconstruction of ventrobasal complex neurons of the thalamus or even regions of dendritic arbors, the degree of convergence is likely to be significantly underestimated. These findings indicate that the anatomical basis exists for an interaction between nociceptive and non-nociceptive somesthetic systems at the level of single ventrobasal neurons of the thalamus of the rat.

Animals