Intraduodenal serotonin elicits non-propagating spike potentials in the small intestine of the rat

Serotonin (5-hydroxytryptamine, 5-HT) is an endogenous signalling molecule capable of altering small intestinal motility. Serotonin is normally present in the intestinal lumen and released by enterochromaffin cells of the mucosal epithelium. We found that intraduodenal infusion of exogenous serotonin causes a dose-dependent myoelectric response in the smooth muscle of the small intestine in the conscious rat. The response consists of repetitive bursts of action potentials (RBAP) that are characterized as short bursts of non-propagative myoelectric spiking. RBAP occur intermittently and only during the first 15 min after intralumenal serotonin infusion. After the initial 15 min period, the frequency of RBAP declines, and the myoelectric pattern shifts to prolonged and continuous spiking, eliminating the interdigestive migrating myoelectric pattern. The effects of intralumenal serotonin are not replicated by parenteral or intraperitoneal infusion nor by intralumenal infusion of 5-hydroxytryptophan or 5-hydroxyindoleacetic acid. The response to intralumenal serotonin was eliminated by several specific 5-HT receptor antagonists. On repeated intralumenal administration of serotonin, the RBAP response decreased demonstrating a decreased sensitivity of the muscle contraction on re-exposure to serotonin. We conclude that intralumenal infusion of serotonin can temporarily initiate specific small intestinal muscle events that are not generated by serotonin from other non-lumenal administration sites. We speculate that an afferent neuro-pathway is necessary for the induction of RBAP, since RBAP are not observed from in vitro muscle preparations.     

Myoelectric response of the small intestine to the orad presence of the tapeworm Hymenolepis diminuta.

During its 24-hr migratory cycle in the small intestine, Hymenolepis diminuta is located in the orad part of the small intestine during the early morning hours and then in the caudad part of the small intestine during the late afternoon and early evening. During the later period, tapeworm-induced alterations of interdigestive myoelectric activity, a correlate of smooth muscle contraction or intestinal motility, are most intense in the ileal region. The hypothesis tested was that the tapeworm-induced changes in intestinal motility are local responses of the intestine responding to the close proximity of the lumenally positioned tapeworm and to the nutritional state of the host. The small intestine was monitored before and for 20 days after infection using electrodes implanted on the serosa of the small intestine. Myoelectric recordings were analyzed for the frequency of the normal patterns of interdigestive myoelectric spiking patterns and the altered myoelectric spiking related to tapeworm infection. During the morning hours, when the tapeworms are situated in the orad small intestine, no changes were observed during the normal myoelectric pattern of the digestive phase in any region of the intestine. When examined after the conversion of the digestive to interdigestive phase of motility, only on day 10 postinfection was the interdigestive phase significantly altered. It was concluded that the presence of the tapeworm in the orad small intestine during the satiety stage of the rat causes no changes in the electric events of the small intestine, with the exception of day 10 postinfection. Because tapeworms in the orad small intestine do not induce the tapeworm-altered myoelectric activity observed in the afternoon and evening with caudally positioned tapeworms, tapeworm-altered motility is not simply a response of the small intestine to the local presence of the tapeworm.     

Hymenolepis diminuta: praziquantel removal of adult tapeworms is followed by apoptotic down-regulation of mucosal mastocytosis.

The rat tapeworm, Hymenolepis diminuta, induces mastocytosis, hypertrophy of enteric smooth muscle, alteration of enteric myoelectric activity, and slowed enteric transit of the rat host's intestine. This report examines the resolution of both tapeworm-induced mastocytosis and tissue changes during the period following removal of the tapeworm with Praziquantel (PZQ). The dynamics of the mucosal mast cell (MMC) population following removal of the tapeworms was assessed by histochemical identification of MMC and morphometric techniques. As a possible mechanism of MMC population regulation, MMC apoptosis was examined over the same experimental period using the in situ nick end labeling of fragmented DNA (TUNEL). Shifts in MMC numbers were correlated with functional and morphological changes of the intestine following removal of the adult-stage tapeworm. Ileal tissues from rats infected 32 days with H. diminuta (the beginning of plateau phase of tapeworm-induced chronic mastocytosis) were harvested 1, 2, 3, and 4 weeks after the PZQ treatment. Control ilea were obtained either from rats which were never infected and never treated with PZQ or from rats infected with H. diminuta for 32 days but not treated with PZQ. In order to detect MMC and apoptosis, tissue sections of ileum were doubled stained sequentially with Astra blue for MMC granules followed by a modification of the TUNEL technique. No alteration in MMC numbers were observed in PZQ-treated animals until 3 weeks after the removal of the tapeworms. The decline of MMC occurred in the mucosa and submucosa. MMC numbers first approached uninfected control levels at 4 weeks posttreatment. Coincident with the decline in mucosal MMC numbers, the rate of MMC entering apoptosis also declined. Simultaneously, ileal smooth muscle layers, hypertrophied by infection, and mucosal structures began the process of involution and atrophy. Apoptosis of MMC in the submucosa and muscularis mucosa was not detected. In conclusion, H. diminuta-elicited mastocytosis and increased thickness of both mucosa and muscularis externa do not begin a decline toward control values until 3 weeks after the parasites are gone and normal intestinal motility is restored. These data are consistent with the lack of MMC mediation of altered motility, and the decline in the rate of MMC apoptosis at 3 weeks post-PZQ suggests that apoptosis may play an important role in the involution of tapeworm-induced mastocytosis.     

Modulation of caudal intestinal permeability in the rat during infection by the tapeworm Hymenolepis diminuta.

Bidirectional movement of solutes between the intestinal lumen and systemic circulation is restricted by tissue barriers that may be altered under conditions such as intestinal infection. In a study using an in vitro everted sac preparation to assess small intestinal permeability in a lumen-to-serosa direction, 51Cr-EDTA movement was compared regionally in the jejunum and ileum of rats infected and uninfected by tapeworms. Whereas jejunal segments showed no significant differences in permeability to 51Cr-EDTA at 6, 15, or 32 days postinfection (dpi), ileal segments displayed an increased permeability on 15 and 32 dpi, but not 6 dpi. The alterations in permeability were not reversed 1 wk after removal of the tapeworm from the intestine. In conclusion, the strictly lumen-dwelling tapeworm infection allows increased movement of molecules from the lumen into ileal, but not jejunal, tissues by 15 dpi.     

Effects of tapeworm infection on male reproductive success and mating vigor in the red flour beetle,Tribolium castaneum

Parasites may exert negative effects on host survivorship and reproductive success. The effects of parasites on female host fitness have been well documented; however, the effects of parasites on the reproductive success of male hosts and particularly the underlying mechanisms that alter male fitness are not well understood. Previous studies demonstrated that infection by rat tapeworm (Hymenolepis diminuta) reduced the fitness of male red flour beetles (Tribolium castaneum) in an environment of female mate choice and strong male-male competition. The present study determined the role of female mate choice and male insemination capacity on observed fitness reduction of male beetles by the tapeworm parasites. We found that infected males showed reduced mating vigor and consequently inseminated fewer females than did uninfected males. Specifically, tapeworm infection reduced the number of offspring sired by a male by 14-22% even when male-male competition and female mate choice were absent. Further, the insemination capacity of males diminished by 30% because of infection. Female beetles did not discriminate against infected males in precopulatory mate choice experiments. Copulatory courtship, a determinant of postcopulatory female choice, was not significantly different between infected and uninfected males. Hence, we concluded that female beetles did not show either pre- or postcopulatory choice against tapeworm-infected males. Therefore, tapeworm-induced reduction in the reproductive success of male beetles possibly results from altered reproductive biology, such as lower mating vigor and decreased sperm quantity or quality.     

Influence of microbial species on small intestinal myoelectric activity and transit in germ-free rats

The effect of an intestinal microflora consisting of selected microbial species on myoelectric activity of small intestine was studied using germ-free rat models, with recording before and after specific intestinal colonization, in the unanesthetized state. Intestinal transit, neuropeptides in blood (RIA), and neuromessengers in the intestinal wall were determined. Clostridium tabificum vp 04 promoted regular spike burst activity, shown by a reduction of the migrating myoelectric complex (MMC) period from 30.5 +/- 3.9 min in the germ-free state to 21.2 +/- 0.14 min (P < 0.01). Lactobacillus acidophilus A10 and Bifidobacterium bifidum B11 reduced the MMC period from 27.9 +/- 4.5 to 21.5 +/- 2.1 min (P < 0.02) and accelerated small intestinal transit (P < 0.05). Micrococcus luteus showed an inhibitory effect, with an MMC period of 35.9 +/- 9.3 min compared with 27.7 +/- 6.3 min in germ-free rats (P < 0.01). Inhibition was indicated also for Escherichia coli X7 gnotobiotic rats. No consistent changes in slow wave frequency were observed. The concentration of neuropeptide Y in blood decreased after introduction of conventional intestinal microflora, suggesting reduced inhibitory control. Intestinal bacteria promote or suppress the initiation and aboral migration of the MMC depending on the species involved. Bacteria with primitive fermenting metabolism (anaerobes) emerge as important promoters of regular spike burst activity in small intestine.     

Effect of size of Trichinella spiralis (Nematode) infections on glucose and ion transport in the rat intestine

An in vivo perfusion technique, using 3 intestinal loops representing the anterior, mid and posterior regions of the rat small intestine, was used to determine intestinal glucose uptake 5 days after infection with Trichinella spiralis. At high levels of infection (3,000 and 6,000 larvae/rat) net glucose absorption by the intestinal mucosa was significantly impaired in all regions of the small intestine when compared to uninfected controls. At low levels of infection (50 larvae/rat) glucose uptake by the mucosa was significantly enhanced in all 3 regions of the small intestine. Intermediate levels of infections (200-1,000 larvae/rat) also enhanced glucose uptake, but only in the anterior regions of the small intestine. When washings from the small intestine of rats infected with 50 larvae/rat were added to the perfusion fluid used on uninfected rats, glucose uptake was also significantly enhanced. These results suggest that at low levels of infection the intestinal lumen contains a metabolite which may affect the mucosal transport of glucose and the related fluxes of H2O, Na+, Cl-, and K+, in the rat intestine. Luminal [H+] and pCO2 decreased from the proximal to distal regions of the small intestine following perfusion; pO2 was significantly decreased in the proximal and distal regions.     

Pathophysiology of cestode infections: Effect of Hymenolepis diminuta on oxygen tensions,pH and gastrointestinal function

Studies on the normal and parasitized rat intestine were used to investigate the effect of the tapeworm, Hymenolepis diminuta, on in vivo intestinal lumenal oxygen tensions, acid-base balance and mucosal absorption and accumulation of fluid and glucose.The lumenal bulk aqueous phase is considerable, well mixed and aerobic with an oxygen tension of 40–50 mm Hg. Neither the unstirred layers adjacent to the brush border membrane nor the area adjacent to the mucosa (“paramucosal lumen”) are significant barriers to the diffusion of oxygen from the blood to the intestinal lumen. In the uninfected distal ileum and colon anoxic conditions may occur in the central lumen, but, in the parasitized intestine fluid absorption is reduced and anoxic conditions do not occur. Increased H⁺ ion concentration in the parasitized intestine plays a role in increasing the availability of oxygen to intestinal helminths. Concomitant with the lower pH, the pCO₂ in the lumen of the parasitized intestine was twice as high as that found in normal animals. The total CO₂ in the parasitized intestine steadily decreased over a 3-h perfusion period, while in the normal intestine the total CO₂ content increased after an initial fall during the first 30 min of perfusion. When the worms were removed, the ability of the intestine to restore normal acid-base balance was restored. Glucose and fluid absorption in both the infected and uninfected intestine were reduced by an increase in H⁺ ion concentration; both parameters were lower in the parasitized intestine than in the normal animals. Low pH increased fluid and glucose transport by H. diminuta.While the dry weights of both the parasitized and uninfected total small intestine and of the intestinal mucosa were the same, the wet weights were considerably different, indicating defective fluid balance in the infected intestine. Accumulation of glucose by the parasitized mucosa was greater than in control animals and decreased with an increase in H⁺ ion concentration. The glucose transport system in the parasitized gut was therefore affected at two levels, one at the brush border, where transport into the mucosa was decreased by lowering the pH, and secondly at the level of the basal and lateral membranes, where transport out of the mucosal tissue into the circulatory system was also reduced.The above results are discussed in terms of current widely accepted but erroneous concepts relating to the intestinal ‘microcosm’.     

Hormone induced changes in lactase glycosylation in developing rat intestine

Lactase exists in both soluble and membrane-bound forms in suckling rat intestine. The distribution of lactase and its glycosylated isoforms in response to thyroxine or cortisone administration has been studied in suckling rats. 75% of lactase activity was detected, associated with brush borders, compared to 24% in the soluble fraction of 8-day-old rats. Thyroxine treatment enhanced soluble lactase activity to 34%, whereas particulate fraction was reduced to 67% compared to controls. Cortisone administration reduced soluble lactase activity from 24% in controls to 12% with a concomitant increase in membrane-bound activity to 89%. Western blot analysis revealed lactase signal, corresponding to 220 kDa in both the soluble and membrane fractions, which corroborated the enzyme activity data. The elution pattern of papain solubilized lactase from agarose-Wheat Germ agglutinin, or Concanavalin A or Jacalin agglutinin columns was different in the suckling and adult rat intestines. Also the elution profile of lactase activity from agarose-lectin columns was modulated in cortisone, thyroxine, and insulin injected pups, which suggests differences in glycosylated isoforms of lactase under these conditions. These findings suggest the role of these hormones in inducing changes in lactase glycosylation during postnatal development of intestine, which may contribute to adult-type hypolactasia in rats.     

The relation of the soluble thiamine triphosphatase activity of various rat tissues to nonspecific phosphatases

Polyacrylamide gel electrophoresis was used to investigate the relation of the soluble thiamine triphosphatase activity of various rat tissues to other phosphatases. This technique separated the thiamine triphosphatase of rat brain, heart, kidney, liver, lung, muscle and spleen from alkaline phosphatase (EC 3.1.3.1), acid phosphatase (EC 3.1.3.2) and other nonspecific phosphatase activities. In contrast, the hydrolytic activity for thiamine triphosphate in rat intestine moved identically with alkaline phosphatase in gel electrophoresis. Thiamine triphosphatase from rat liver and brain was also separated from alkaline phosphatase and acid phosphatase by gel chromatography on Sephadex G-100. This gave an apparent molecular weight of about 30,000 and a Stokes radius of 2.5 nanometers for brain and liver thiamine triphosphatase. The intestinal thiamine triphosphatase activity of the rat was eluted from the Sephadex G-100 column as two separate peaks (with apparent molecular weights of over 200,000 and 123,000) which exactly corresponded to the peaks of alkaline phosphatase. The isoelectric point (pI) of the brain thiamine triphosphatase was 4.6 (4 degrees C). The partially purified thiamine triphosphatase from brain and liver was highly specific for thiamine triphosphate. The results suggest that, apart from the intestine, the rat tissues studied contain a specific enzyme, thiamine triphosphatase (EC 3.6.1.28). The specific enzyme is responsible for most of the thiamine triphosphatase activity in these tissues. Rat intestine contains a high thiamine triphosphatase activity but all of it appears to be due to alkaline phosphatase.     

Intestinal immunity suppresses carrying capacity of rats for the model tapeworm, Hymenolepis diminuta

Hymenolepis diminuta is a parasitic tapeworm of the rat small intestine and is recognized as a useful model for the analysis of cestode-host interactions. In this study, we analyzed factors affecting the biomass of the tapeworm through use of rat strains carrying genetic mutations, namely X-linked severe combined immunodeficiency (xscid; T, B and NK cells deficiency), nude (rnu; T cell deficiency), and mast cell deficient rats. The worm biomass of F344-xscid rats after infection with 5 cysticercoids was much larger than control F344 rats from 3 to 8?weeks. The biomass of F344-rnu rats was also larger than the controls, but was intermediate between F344-xscid and control rats. These observations demonstrated that host immunity can control the maximal tapeworm biomass, i.e., carrying capacity, of the rat small intestine. Both T cell and other immune cells (B and NK cells) have roles in determining the carrying capacity of tapeworms. Total worm biomass and worm numbers in mast cell deficient rats (WsRC-Ws/Ws) were not significantly different from control WsRC-+/+ rats after 3 and 6?weeks of primary infection. Mast cell deficient rats displayed reinfection resistance for worm biomass but not worm expulsion. These findings suggest that the mast cell has a role for controlling the biomass of this tapeworm in reinfection alone, but does not affect the rate of worm expulsion. Overall, our findings indicate that the mast cell is not a major effector cell for the control of the carrying capacity of tapeworms. The identity of the major effector cell remains unknown.     

Tachykinin-stimulated small bowel myoelectric pattern: sensitization by NO inhibition, reversal by neurokinin receptor blockade

Tachykinins stimulate motility whereas NO inhibits motility in the gastrointestinal tract. AIM: To investigate if inhibition of NO production sensitizes myoelectric activity to subthreshold doses of tachykinins in the small intestine of awake rats. METHODS: Rats were supplied with a venous catheter and bipolar electrodes at 5, 15 and 25 cm distal to pylorus for electromyography of small intestine. The motor responses were evaluated using pattern recognition. Substance P and neurokinin A dose-dependently stimulated gut motility, with neurokinin A being more potent than substance P. Therefore, neurokinin A was chosen and administered under baseline conditions and 45-60 min after N(omega)-nitro-L-arginine (L-NNA) 1 mg kg(-1), with or without pretreatment with L-arginine 300 mg kg(-1). In addition, myoelectric activity effects of neurokinin A in conjunction with L-NNA were studied before and after administration of the tachykinin receptor antagonists, SR140333 (NK1), SR48968 (NK2) and SR142801 (NK3), each at 2.5 mg kg(-1). RESULTS: Dose-finding studies verified 10 pmol kg(-1) min(-1) to be the threshold dose at which NKA caused phase II-like activity in a low percentage of experiments (12%, n=41). This dose was therefore used in combination with L-NNA for sensitization experiments of gut myoelectric activity. In experiments where NKA-induced no response, pretreatment with L-NNA led to phase II-like activity in 9 of 18 (50%, p<0.05) experiments. Co-administration of SR140333 and SR48968 abolished this effect. CONCLUSION: NO counteracts the stimulatory effect of tachykinins on small bowel myoelectric activity in the rat. Inhibition of the L-arginine/NO pathway sensitizes the gut to tachykinin-stimulated motor activity.     

Effect of cloned Salmonella typhimurium enterotoxin on rabbit intestinal motility

Abstract We analysed the small intestine myoelectric responses of anesthetized New Zealand albino rabbits to Escherichia coli lysates containing an enterotoxin cloned from Salmonella typhimurium . Migrating action potential complex, which consisted of rapid bursts of action potentials and secretion of fluid, was observed only in ileal loops injected with the enterotoxin-containing lysate. Migrating action potential complex produced by Stn usually propagated aborally, which was typical of cholera toxin, but orad or bidirectional propagation occurred from a single point of origin when activity was intense. Gell lysates from an E. coli clone containing vectors alone, as well as proximal control segments injected with phosphate-buffered saline, gave neither a change in motility nor fluid secretion. These results show that Stn caused dramatic changes in intestinal motility and substantial fluid production.     

The spatial and temporal concentrations of choline in the lumen contents of the small intestine of uninfected and Hymenolepis diminuta infected rats

The spatial and temporal concentrations of free choline in the lumen of the small intestine of the uninfected and Hymenolepis diminuta-infected rat were investigated. In the unfed infected or uninfected rat, the choline concentrations ranged from approximately 500 microM in the duodenum to approximately 20 microM in the posterior ileum, with some segments in infected rats containing significantly higher choline levels than in uninfected rats. Following feeding, choline levels were significantly elevated to approximately 3 mM by 6 h, although these concentrations fell rapidly in transit down the intestine. By 12 h the choline levels were similar to those in the unfed rat. An initial small shift in the worm biomass toward the duodenum after feeding was followed by a redistribution of biomass along the length of the small intestine. The worm biomass, however, had little or no effect on the choline levels. The high concentrations of free choline observed in the anterior regions of the intestine are postulated to be predominantly determined by nutritional intake while the concentrations in the posterior region may in part be determined by blood choline levels. The high levels of choline indicate that choline is not limiting to support the growth of the worms.     

Peculiarities of protein hydrolysis on the digestive-transport surfaces of the intestine of the Kittiwake Rissa Tridactyla and Alcataenia Larina (Cestoda, Dilepididae) parasitizing it

Research of the activity of proteolytic enzymes functioning on the digestive-transport surfaces of the intestine of infected and noninfected kittiwakes Rissa tridactyla and cestodes Alcataenia larina (Cestoda, Dilepididae) parasitizing it was conducted. The protease activities depending on the pH values in the intestine of gulls and tapeworms were determined. The dynamics of desorption of enzymes of protein hydrolysis from the surface of the cestodes A. larina and the kittiwake intestine was studied. It was shown that the tapeworm invasion activates the processes of membrane and cavitary digestion of birds. It was determined that the degree of influence of the dilepidid infection on protease activity was defined mainly by the intensity of the invasion.     

Myoelectric signal processing for control of powered limb prostheses

Progress in myoelectric control technology has over the years been incremental, due in part to the alternating focus of the R&D between control methodology and device hardware. The technology has over the past 50 years or so moved from single muscle control of a single prosthesis function to muscle group activity control of multifunction prostheses. Central to these changes have been developments in the means of extracting information from the myoelectric signal. This paper gives an overview of the myoelectric signal processing challenge, a brief look at the challenge from an historical perspective, the state-of-the-art in myoelectric signal processing for prosthesis control, and an indication of where this field is heading. The paper demonstrates that considerable progress has been made in providing clients with useful and reliable myoelectric communication channels, and that exciting work and developments are on the horizon.     

Myeloelectric complex of the small intestine and variations in the intestinal motility in sheep]

The motor pattern of the small intestine of the sheep fed ad libitum is characterized by the regular occurrence of myoelectric complexes comprising a phase of regular and irregular spiking activities. Each complex is propagated along the ovine small intestine at a mean velocity of 17 cm/min and originated on the duodenal bulb at 70-100 mn intervals. Reduction and increase in duration of the phase of irregular spiking activity of the complex occurred during fasting and overfeeding respectively. Reduction in spiking activity is paralleled by an increased velocity of propagation whilst a lower migration and a reduced number of the complexes are characteristic of overfeeding. It is concluded that duration, number and velocity of propagation of the myoelectric complexes are adaptative factors in changes of the intestinal flow rate.     

The spatial and temporal concentrations of choline in the lumen contents of the small intestine of uninfected and Hymenolepis diminuta infected rats

The spatial and temporal concentrations of free choline in the lumen of the small intestine of the uninfected and Hymenolepis diminuta-infected rat were investigated. In the unfed infected or uninfected rat, the choline concentrations ranged from 500 μM in the duodenum to 20 μM in the posterior ileum, with some segments in infected rats containing significantly higher choline levels than in uninfected rats. Following feeding, choline levels were significantly elevated to approximately 3 mM by 6 h, although these concentrations fell rapidly in transit down the intestine. By 12 h the choline levels were similar to those in the unfed rat. An initial small shift in the worm biomass toward the duodenum after feeding was followed by a redistribution of biomass along the length of the small intestine. The worm biomass, however, had little or no effect on the choline levels. The high concentrations of free choline observed in the anterior regions of the intestine are postulated to be predominantly determined by nutritional intake while the concentrations in the posterior region may in part be determined by blood choline levels. The high levels of choline indicate that choline is not limiting to support growth of the worms.     

Effect of surgical alteration of the rat gastrointestinal tract on the growth and development of Hymenolepis diminuta

Eight groups of rats were used to study the involvement of the enteric (ENS) and central (CNS) nervous systems in the development of Hymenolepis diminuta using surgical intestinal transection, or CNS denervation, or both procedures. The transection procedure was used to isolate the ENS of the small intestine from either orad and/or caudal portions of the alimentary system, while the CNS denervation was used to eliminate direct visceral efferent inputs from the CNS. Nine days after the surgical procedures, all rats were infected with 35 cysticercoids of H. diminuta. On 20 days postinfection, the infection intensity, tapeworm dry weight, tapeworm morphology, intestine length, and intestinal wet weight were recorded. Only the combination of the duodenal and ileal transections with a CNS denervation reduced infection intensity and prevented the increased intestinal length normally observed in infected rats. In contrast, none of the various intestinal transection procedures alone or CNS denervation alone had any effect on the survival, ability to produce oncospheres or morphology of the tapeworms. In conclusion, tapeworm survival is decreased when both CNS and ENS inputs into the small intestine are altered or absent.