Biomedical subjects

T K Smith

Publications and source records attributed to T K Smith.

At least 127 records · Page 7Linked to original sources

PubMed1989

Effects of dietary polyamine precursors on the metabolism and tissue concentrations of amino acids in the rat.

Experiments were conducted to determine the effects of dietary supplements of amino acid precursors of polyamines on amino acid metabolism in the rat in order to better understand comparative aspects of polyamine metabolism. Rats were fed isonitrogenous combinations of methionine, ornithine, arginine and 2-difluoromethylornithine in casein-based diets. It was observed that hepatic concentrations of methionine, 5'-deoxy-5'-methylthioadenosine and decarboxylated S-adenosylmethionine were more easily influenced by diet than were arginine and metabolites. It was concluded that rats may be more refractory to exogenous polyamine precursor amino acids than are chicks because of the presence of a functional urea cycle.

Adenosine↗

PubMed1989

Histological identification of the interstitial cells of Cajal in the guinea-pig small intestine.

In order to clarify the contraversial structure that Cajal (1889, 1893, 1911) called "interstitial cells" in the intestine, we have compared the descriptions by Cajal with our recent findings. Cajal defined three groups of interstitial cells in the intestine: those in the mucosa, the deep muscular plexus, and the myenteric plexus. In the deep muscular plexus, he described cells with small perikarya and long, branching processes. Neither glial cells nor ZIO positive fibroblast-like cells, the only cell types seen in the numbers and location necessary to support Cajal's observations, conformed with the morphology he described. Cajal might have described a composite cell, or chimera, the cell body being that of a glial (or perhaps a fibroblast-like) cell and the processes being simultaneously stained neurites that ran in close association with the cell. In the myenteric plexus, staining with S-100 protein for glial cells and ZIO for fibroblast-like cells reveal cells between the external muscle layers. Cells stained with S-100 protein resemble the drawings and also his description of the interstitial cells, although these cells appear to be less frequent than Cajal's drawings indicate. There are numerous glial cells in the ganglia and primary strands of the plexus which were not included in Cajal's publications. Conversely, the fibroblast-like cells are incidentally associated with nerve fibers. Although they lie in the same plane, the fibroblast-like cells form a pattern distinct from that of the nerve fibers of the tertiary component of the myenteric plexus. These cells occur in the distribution and numbers of the interstitial cells described by Cajal. They are stained by ZIO and also immunoreactive for gamma-aminobutyric acid. They can be identified by scanning electron microscopy. In the mucosa, Golgi staining reveals a pattern of nerve fiber bundles that is indistinguishable from Cajal's drawings. Glial cell bodies frequently occur at the intersections of these bundles and appear as cell nuclei surrounded by a cytoplasm which is actually the stained nerve fiber bundles. What Cajal depicted as interstitial cells were composite structures consisting of glial cells and contiguous nerve fiber bundles. We conclude that the structures which Cajal called interstitial cells in the intestine do not originate from one cell type. Nevertheless, two groups of fibroblast-like cells, those lying parallel and close to the nerve strands of the deep muscular plexus and those of the myenteric plexus, can be recognized by ZIO staining and scanning electron microscopy.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1989

Correlated functional and structural analysis of enteric neural circuits.

Views about the roles and nature of the enteric nervous system have changed dramatically in the last ten years. This system of neurons is recognized to control, through reflex pathways intrinsic to the gut wall, motility, transport of water and electrolytes, and blood flow in the small and large intestines. There are thus a range of neuron types in the enteric nervous system, including sensory neurons, motor neurons, and interneurons, involved in the control of each of these functions. The present paper deals with the recent efforts to provide an integrated functional and structural description of the nerve circuits. One of the challenges in this quest has been to identify primary sensory neurons and final motor neurons involved in motility control. Evidence is presented that in the guinea-pig small intestine the primary sensory neurons have Dogiel type II morphology and are, electrophysiologically, AH neurons. They send circumferential processes to adjacent myenteric ganglia and some of them, at least, have processes leading from the mucosa. Motor neurons are S neurons, with cell bodies in the myenteric plexus. Those that supply the circular muscle are Dogiel type I neurons and provide processes that run circumferentially for about one third of the circumference of the intestine. The circuits for secretomotor reflexes have been partly worked out. The secretomotor neurons have cell bodies in the submucous ganglia and are activated to return water and electrolytes to the lumen during digestion. The secretomotor reflexes are regulated, in accord with whole body water and electrolyte homeostasis, via sympathetic neurons which lower the excitability of secretomotor neurons.

Animals↗

PubMed1988

Reflex changes in circular muscle activity elicited by stroking the mucosa: an electrophysiological analysis in the isolated guinea-pig ileum.

A preparation of isolated small intestine from the guinea-pig was studied in which reflex responses of the circular muscle were recorded intracellularly when sensory receptors in the mucosa were stimulated mechanically. This preparation was used to examine the properties of mucosa to muscle reflexes that involve non-cholinergic motor neurons innervating the circular muscle. Reproducible stimulation of the mucosa was achieved by stroking with a motor-driven brush. Gentle brush-strokes applied to the mucosa typically evoked inhibitory junction potentials anal to the stimulus and excitatory junction potentials at recording sites oral to the stimulus. Both events were rapid in onset and up to 25 mV in amplitude. The reflexes were blocked by tetrodotoxin (0.5 microM). Junction potentials declined in amplitude with distance from the stimulus, the amplitude of the excitation 15 mm oral to the stimulus was half that at 5 mm from the stimulus, whereas the amplitude of the inhibitory potential at 40-45 mm was about 60% of that at 5-10 mm anal to the stimulus. Hexamethonium (100-200 microM) blocked the ascending excitation but only slightly reduced the descending inhibition. Ascending excitation was blocked by antagonists for substance P receptors in the muscle, and inhibition was substantially reduced by apamin (0.2 microM), both before and after hexamethonium. Both responses were abolished by removal of the mucosa from the stimulus site and when lesions were made through the myenteric plexus between the stimulation and recording sites, but persisted when similar lesions were made through the submucous plexus. It is concluded that there are neurons with mechanoreceptive nerve endings in the mucosa. Stimulation of such sensory neurons leads to activation of pathways in the myenteric plexus that excite motor neurons to the muscle both oral and anal to the stimulation site. The demonstration that mucosa to muscle reflexes can be consistently evoked in the small intestine in vitro provides an opportunity for close analysis of the reflex pathways. Such analysis is not so readily achieved when reflexes are initiated by distension that, by moving the intestine, can dislodge the recording electrode.

Animals↗

PubMed1988

An electrophysiological study of the projections of motor neurones that mediate non-cholinergic excitation in the circular muscle of the guinea-pig small intestine.

The projections of neurones that produce the fast non-cholinergic excitatory junction potentials (e.j.p.s) in the circular muscle were analysed in the isolated ileum of the guinea-pig. Standard intracellular microelectrode techniques were used to record the amplitudes of such e.j.p.s in response to short trains of stimuli from transmural electrodes. Projections of the neurones around the circumference of the intestine were determined by plotting the change in e.j.p. amplitude with distance from longitudinally placed electrodes. Projections in the oral and anal directions were examined by recording at varying distances from transversely placed electrodes, and also by recording responses elicited close to longitudinal electrodes at various distances from lesions made 3-5 days earlier to interrupt orally and anally directed pathways. Experiments were performed in the presence of hexamethonium to determine the projections of the final motor neurones and in the absence of the drug to examine the projections of excitatory inputs to these neurones. With hexamethonium present, there was a decline in e.j.p. amplitude to 7.5% of maximum at 12 mm (a half circumference) from longitudinal stimulating electrodes. The decline was much less if hexamethonium was not present, and slightly greater if the myenteric plexus was removed. Thus, excitatory motor neurones and cholinergic neurones that impinge upon them both project circumferentially. When the longitudinal muscle and myenteric plexus were removed, and 3-5 days allowed for terminals to degenerate, no e.j.p.s could be recorded in the circular muscle, indicating that the fibres reach the circular muscle from the myenteric plexus. Following transverse lesions, substantial deficits in excitatory transmission only occurred within 1 mm oral or anal to the lesions indicating that the majority of neurones have only short projections along the intestine. A slight deficit in e.j.p. amplitude, up to 20%, was observed extending to about 5 mm oral; but normal transmission was restored by about 10 mm for the lesions. Thus there is a minority of excitatory motor neurones with oral projections up to about 10 mm in length. Results with transmural stimulation showed that these nerve fibres can cause excitation both when orthodromically and when antidromically stimulated, indicating that they provide collaterals along their lengths. With no hexamethonium present, e.j.p.s exhibited little decrement in amplitude over distances of over 30 mm oral or anal indicating that there are both ascending and descending cholinergic pathways that impinge on the final motor neurones.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

PubMed1988

A role for tryptophan in regulation of protein synthesis in porcine muscle.

Experiments were conducted to determine the effect of varying concentrations of dietary tryptophan on growth rate and protein synthesis in edible muscle tissues of growing swine. A total of 45 immature swine (initial weight approximately 24 kg) were fed corn-gelatin diets containing 0.5 (n = 8), 0.8 (n = 10), 1.3 (n = 10), 1.5 (n = 7) or 2.0 (n = 10) g tryptophan/kg diet for 35 d. Animals fed 0.5 and 0.8 g tryptophan/kg grew more slowly, consumed less feed and had a lower efficiency of feed utilization than animals fed higher concentrations of tryptophan. Thirty similar animals were used in a second experiment. Diets containing 0.5, 0.8, 1.0, 1.5 or 2.0 g tryptophan/kg diet (n = 6) were fed for 14 d, after which all animals were killed and samples were taken of longissimus dorsi, triceps brachii and biceps femoris. Protein synthetic activity was determined by monitoring the incorporation of [14C]phenylalanine into protein in vitro. There was no significant difference in synthetic activity between different muscle types. There was no effect of diet on the activity of the muscle soluble protein fraction. The activity of the muscle ribosomal fraction, however, was positively correlated with increasing concentrations of dietary tryptophan. It was concluded that tryptophan has the potential to regulate muscle protein synthesis in a manner beyond serving simply as a component of protein.

Administration, Oral↗

PubMed1988

Potential for dietary amino acid precursors of neurotransmitters to overcome neurochemical changes in acute T-2 toxicosis in rats.

Experiments were conducted to determine the potential for overcoming T-2 toxin-induced changes in brain neurotransmitter concentrations through dietary manipulation. Rats were fed either a tryptophan-deficient, gelatin-based diet or the same diet supplemented with a mixture of large neutral amino acids for 4 d. Rats were then dosed with 0 or 2.0 mg T-2 toxin/kg body weight and killed 4, 8 or 12 h after dosing. The large neutral amino acid supplements successfully reduced brain concentrations of tryptophan and serotonin in control rats, but this was not enough to overcome the acute effects seen in T-2 toxin-treated rats. A further experiment was then conducted to monitor the effect of T-2 toxin on the ratio of free to protein-bound tryptophan in plasma. Total plasma tryptophan increased in T-2 toxin-treated rats, although there were no significant differences in the ratio of free to protein-bound tryptophan. A final experiment was conducted to determine the specificity of the T-2 toxin effect on concentrations of plasma amino acids. Concentrations of amino acids that use the large neutral amino acid transport system into the brain were higher in T-2 toxin-treated animals. The only other amino acid that had a higher concentration was arginine. It was concluded that acute doses of T-2 toxin may selectively alter membrane transport of amino acids.

Amino Acids↗

PubMed1988

Effect of dietary protein deficiency and L-2-oxothiazolidine-4-carboxylate on the diurnal rhythm of hepatic glutathione in the rat.

Maximizing hepatic glutathione (GSH) concentration may provide greater protection against toxic compounds. A dietary supplement of L-2-oxothiazolidine-4-carboxylate (OTC), a stable derivative of cysteine, increased hepatic GSH in rats fed for 2 wk a diet deficient in protein (7.5%) but not in rats fed a diet adequate in protein (15%). Experiment 2 determined whether a dietary supplement of OTC could maintain the maximum GSH concentration over 24 h. Rats acclimatized for 5 d to a 7.5% protein diet and then fed a 7.5% protein diet supplemented with either 2.5 mmol OTC or cysteine-HCl (CYS)/100 g diet had a more rapid increase in hepatic GSH (4 and 8 h after beginning of feeding, P less than 0.05) than did rats fed an unsupplemented 7.5% protein diet. This response was not due simply to the greater supply of cysteine for GSH synthesis because supplementing the 15% protein diet with OTC or CYS had no effect on the hepatic GSH of rats acclimatized to a 15% protein diet. In experiment 3, rats acclimatized to the 7.5% protein diet had a more rapid rate of increase in hepatic GSH concentration in response to feeding than did rats acclimatized to a 15% protein diet, regardless of which diet they were fed during the 24-h period. It was concluded that in addition to cysteine availability, previous dietary protein status plays a key role in the regulation of the feeding-induced diurnal rhythm of hepatic GSH concentration in rats.

Analysis of Variance↗

PubMed1988

Evaluation of six screening methods for detecting significant bacteriuria.

Six screening methods for the successful detection of significant bacteriuria--electrical impedance (Malthus), automated acridine-orange staining (Autotrak), particle counting (Ramus), bioluminescence, nitrite and leucocyte test strip (BM Nephur), and microscopy--were evaluated. All had excellent predictive values for a negative result (97%-100%) but were less accurate in predicting a positive result (31%-83%). All methods had high sensitivities (83%-100%) but lower levels of specificity (68%-79%). Bioluminescence was the method with the highest specificity (79%) and the lowest rate of false positive results (15%). It would be inappropriate to decide on treatment and management on the basis of the positive results achieved with any of the methods evaluated, but all methods tested could be used for screening out negative results.

Bacteriological Techniques↗

PubMed1988

The effect of dietary protein and sulfur amino acids on hepatic glutathione concentration and glutathione-dependent enzyme activities in the rat.

Hepatic glutathione concentration and glutathione-dependent enzymes, glutathione S-transferase, glutathione peroxidase, and glutathione reductase, are important for protection against toxic compounds. Rats were fed diets containing 4, 7.5, 15, or 45% protein for 2 weeks. Glutathione and cysteine concentrations in rats fed the 4 and 7.5% protein diets were significantly lower (p less than 0.05) than in rats fed the 15 and 45% protein diets. Glutathione S-transferase activity increased with increasing dietary protein. Glutathione peroxidase activity was significantly lower (p less than 0.05) in rats fed 4 and 7.5% protein compared with rats fed 15 and 45% protein, whereas the activity of glutathione reductase was higher in rats fed 4 and 7.5% protein then in rats fed 15 or 45% protein. Dietary sulfur amino acids alone could account for the increase in glutathione concentration resulting from the increase in dietary protein from 7.5 to 15%. The limited availability of glutathione in animals fed the low protein diets could reduce the potential for detoxification of xenobiotics.

Amino Acids, Sulfur↗

PubMed1988

Effects of membrane potential on electrical slow waves of canine proximal colon.

The effects of membrane potential on the waveforms and propagation of slow waves were tested using circular muscles of the canine colon. Studies were conducted with intracellular recording techniques on cross-sectional strips of canine proximal colon. Circular muscle cells near the submucosa generated slow waves that decayed in amplitude as they spread through the circular layer. The membrane potentials of cells were less negative as a function of distance from the submucosal border. Cells near the submucosa were depolarized with elevated external K+ and electrical pulses using the partitioned chamber technique. The waveforms of depolarized submucosal cells were compared with events recorded from cells in the bulk of the circular layer. The waveform changes caused by experimental depolarization were different from the changes in waveform that occur during propagation, suggesting the latter are due to a different mechanism than depolarization. The effects of the membrane potential on syncytial input resistance and length constant were also evaluated. The results of these studies are consistent with the hypothesis that slow-wave propagation across the circular layer in canine proximal colon occurs passively.

Animals↗

PubMed1988

Effect of dietary ornithine on renal and hepatic polyamine synthesis.

Chicks with genetically elevated renal arginase activity were fed crystalline amino acid diets varying in ornithine concentration (0, 1 or 2%) to assess the potential for precursor regulation of polyamine synthesis. Renal arginase and renal and hepatic ornithine decarboxylase activities fell when ornithine was fed. Renal and hepatic ornithine concentrations rose while putrescine concentrations varied quadratically with ornithine feeding. Spermidine and spermine concentrations were not affected by diet. It was concluded that ornithine synthesized in vivo was a more potent stimulator of polyamine synthesis than ornithine of dietary origin.

Animals↗

PubMed1988

Flow cytometry DNA analysis on tumor cell subpopulation of human tumor specimens by exclusion of lymphohemopoietic cells.

We describe a new flow cytometry procedure in which DNA analyses can be obtained selectively on pure, freshly obtained tumor cell subpopulations of human tumor specimens. This procedure is based on exclusion from analysis of the contaminating lymphohemopoietic cells mixed with tumor cells in tumor specimens. This exclusion is made possible by labeling all lymphohemopoietic cells with an antibody to HLe-1 (HLE), which is present on all lymphohemopoietic cells but on no other cells, and by gating against these labeled cells when analyzing for DNA. For the model system, a 1:1 mixture of normal human peripheral blood leukocytes and either of two human cancer cell lines, HEp-2 and MCF-7, normal leukocyte contamination can be reduced to 3.1% while retaining 94.7% of tumor cells for DNA analysis. Four examples of human tumor samples, two cases each of malignant effusions and lymph node metastases, were analyzed with this procedure. The results clearly indicate that this new method will improve ploidy analysis/aneuploidy detection and will make it possible to obtain more accurate cell-cycle analyses of tumor cells than have previously been possible. This new procedure will contribute to clinical and biological studies involving DNA of human tumors.

Adenocarcinoma↗

PubMed1988

Effect of acute oral doses of T-2 toxin on tissue concentrations of biogenic amines in the rat.

T-2 toxin [3 alpha-hydroxy-4 beta, 15-diacetoxy-8 alpha-(3-methylbutyryloxy)-12,13-epoxytrichotec-9-ene] is an emetic Fusarium trichothecene mycotoxin known to cause lethargy, ataxia and feed refusal in economically important animals. Experiments were conducted to determine the effect of acute oral doses of T-2 toxin on tissue concentrations of neurotransmitters thought to play some role in regulation of feed consumption. Sixty-seven male weanling rats were intubated with a few grams of diet in a liquid slurry with or without 2.0 mg T-2 toxin per kilogram of body weight. At 1, 2, 4, 6, 8, 12, 24 and 48 h following dosing, rats were killed, and brains, spleens, hearts and adrenal glands were excised and analyzed for concentrations of neurotransmitters and metabolites using high-performance liquid chromatography with electrochemical detection. Administration of T-2 toxin caused increases in brain concentrations of tryptophan and serotonin at the early time intervals after dosing. Brain concentrations of dopamine increased, whereas concentrations of 3,4-dihydroxyphenylacetic acid (DOPAC) decreased at the later time interals following dosing. Concentrations of dopamine were increased in adrenal glands, whereas epinephrine concentrations decreased. Epinephrine was detected in spleen and heart after administration of T-2 toxin. It was concluded that the increase in brain indoleamines induced by T-2 toxin could contribute to feed refusal in animals suffering from T-2 toxicosis.

Administration, Oral↗

PubMed1988

Regulation of polyamine synthesis by dietary alpha-aminoisobutyric acid and ornithine.

An experiment was conducted to determine the effect of feeding ornithine in combination with alpha-aminoisobutyric acid (AIB), an inhibitor of arginase, on the regulation of polyamine synthesis in chicks. A total of 48 chicks with genetically elevated renal arginase activity was fed diets containing crystalline amino acids and 1% AIB with or without 2% ornithine. Feeding AIB reduced renal arginase activity, while renal and hepatic ornithine decarboxylase (ODC) activity increased. Feeding AIB plus ornithine caused no further reduction in renal arginase activity compared with that in chicks fed the AIB-supplemented diet. Renal and hepatic ODC activities, however, fell to below control levels. Renal, hepatic, and breast muscle ornithine concentrations increased substantially when ornithine was fed. AIB plus ornithine increased renal putrescine and spermidine concentrations. It was concluded that AIB could partially overcome the ornithine-induced inhibition of ODC activity. These findings support the hypothesis that dietary manipulation of precursor amino acids of polyamines in the presence of metabolites that induce ODC activity can influence tissue polyamine concentrations.

Adenosylmethionine Decarboxylase↗

PubMed1987

Effect of dietary lysine on polyamine synthesis in the chick.

Experiments were conducted to determine the effect of dietary lysine on hepatic and renal polyamine synthesis in the chick. High arginine-requiring (HA) and low arginine-requiring (LA) strains were used, with differences in arginine requirements being due, in part, to differences in renal arginase activity. Arginine requirements were 1.2 and 0.9% for the HA strain and LA strain, respectively. A total of 96 chicks (16 per diet, 48 per strain) were fed a crystalline amino acid diet containing requirement concentrations of arginine and 0.95, 1.40 or 1.85% lysine for 2 wk. Hepatic arginase activity was unaffected by dietary lysine in both strains, whereas hepatic ornithine decarboxylase (ODC) activity fell with increased dietary lysine in the LA strain only. Increasing dietary lysine significantly increased renal arginase activity but reduced renal ODC activity in the HA strain only. Increasing dietary lysine also caused decreased renal concentrations of arginine, whereas lysine and ornithine concentration increased in both strains. Concentrations of putrescine also rose. It was concluded that excess dietary lysine increased renal concentrations of ornithine due to induction of arginase. This accumulation of polyamine precursors resulted in increased concentrations of putrescine despite feedback inhibition of ODC.

Adenosylmethionine Decarboxylase↗

PubMed1987

Origin and propagation of electrical slow waves in circular muscle of canine proximal colon.

Experiments to determine the site of slow-wave origin and the mechanism of propagation were performed on muscles of the canine proximal colon. Cells along the submucosal border of the circular layer had resting membrane potentials (RMP) averaging -78 mV, and slow waves, 40 mV in amplitude. The RMP of cells through the thickness of the circular layer decreased exponentially with distance from the submucosal border, such that RMPs of circular cells at the myenteric border were only -43 mV. Slow waves decreased in amplitude through the thickness such that slow waves could not be detected adjacent to the myenteric border. When a thin strip of muscle along the submucosal border was removed, slow waves were not recorded from the bulk of the circular layer and could not be evoked by acetylcholine. Slow waves were still present in the excised strip. Experiments to determine the rate of slow-wave propagation were also performed. Two cells were impaled, one at the submucosal surface, and another at some distance through the circular layer. Slow waves occurred nearly simultaneously at both sites. What latency was observed could be explained on the basis of electrotonic conduction. The results support the hypothesis that in the canine proximal colon slow waves are generated at the extreme submucosal surface of the circular layer. The bulk of the circular layer does not possess either pacemaker or regenerative mechanisms, and slow waves propagate passively toward the myenteric border. The cable properties of the circular muscle syncytium furnish a barrier to invasion of the longitudinal layer by the slow wave event.

Acetylcholine↗

PubMed1987

Interaction of two electrical pacemakers in muscularis of canine proximal colon.

Experiments were performed to determine the source of the 20 cycles/min electrical oscillation commonly seen in colonic electrical records, the influence of the 20 cycles/min rhythm on the circular and longitudinal muscle layers, and the interactions between the 20 cycles/min rhythm and slow waves in circular muscle cells. Cross-sectional muscle preparations of the canine proximal colon were used to allow impalement of cells at any point through the thickness of the muscularis. Intracellular recordings from circular muscle cells clearly showed the two characteristic pacemaker frequencies in the colon (6 cycles/min slow waves; 20 cycles/min oscillations). The 20 cycles/min oscillations were recorded from longitudinal and circular muscle cells. Their amplitudes were greatest at the myenteric border. In the longitudinal layer the 20 cycles/min events initiated action potentials; in circular muscle the 20 cycles/min events summed with slow waves. Simultaneous recordings from circular and longitudinal cells across the myenteric border demonstrated that events in the two layers were usually in phase, suggesting that the two layers are electrically coupled and are paced by a common pacemaker. The amplitude of the 20 cycles/min events decayed with distance from the myenteric border in both circular and longitudinal muscles. The data demonstrate that two discrete populations of pacemaker cells generate the spontaneous electrical activity in the colon. Both events appear to passively spread through the circular muscle. It is the summation of these events that appears to serve as the signal for excitation-contraction coupling in circular muscle.

Animals↗