Proefschrift Maastricht. / Met lit. opg. - Met samenvatting in het Nederlands.

Thesis (M.Sc. (Basic Medical Science))--Mahidol University, 1969.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1959. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 55-59).

No description available.

Infection by <I>Salmonella enteritidis </I>and <I>S. typhimurium</I> has been studied in the rat (Hooded Lister) model <I>in vivo</I>. Salmonellosis in the Hooded Lister rat has many similarities with the disease in humans. <I>Salmonellae</I> associated with the small intestinal epithelium as early as 4 h after intragastric intubation. Therefore, reinfection from the large intestine initially may not play a significant role in the infection process. Both <I>Salmonella</I> serotypes colonised, persisted and proliferated in the gastrointestinal tract and invaded sub-epithelial tissues, mainly via the ileum, leading to the systemic distribution of these pathogens. Coincidental with the infection, polyamine levels and crypt cell proliferation rates in the small intestine increased resulting in substantial growth of the tissue. This growth was particularly dramatic in the ileum where there was also some disruption of the villous epithelium. It is possible that these effects of the infection on the metabolism and morphology of the small bowel, which strongly resembles the changes induced by some plant lectins, may facilitate the colonisation and invasion of the gut by <I>Salmonella sp.</I> It was shown for the first time, that <I>S. enteritidis</I> and <I>S. typhimurium</I> strains expressing type 1 fimbriae closely associated with the rat ileal epithelium <I>in vivo</I>. Moreover, orally administered purified type 1 fimbriae also associated with the ileal surface. Thus, type 1 fimbriae alone or in combination with other fimbriae may play an important role in the early stages of infection with these pathogenic bacteria. This was further supported by the finding that deletion of the type 1 fimbriae from a strain of <I>S. enteritidis</I> led to a reduction in the numbers of <I>Salmonella </I>present in the distal ileum 6 h post-dosing. The deletion of type 1 fimbriae did not however affect the long-term progression of infection, suggesting that type 1 fimbriae do not play a prominent role in persistence.

We investigated the origins, neural modulation, ionic mechanisms, and electrical coupling properties of the pacemaker systems in the canine ileum by simultaneously recording the intracellular electrical activity and accompanying mechanical activity in cross-sectioned slabs of the muscularis extema or in the isolated circular muscle. In the whole thickness preparation, intracellular recordings were taken from the circular muscle near the myenteric plexus (MyP), deep muscular plexus (DMP), and intermediate areas between the MyP and DMP and in the isolated circular muscle preparation from similar areas except near the myenteric plexus. One type of slow wave, sigmoidal or triangular in shape, was recorded from impalements near the DMP region in the whole-thickness preparation. Another type observed from the MyP region oscillated at nearly the same frequency (9-10 cycles/min) and was characterized by a fast upstroke and a square shape. A mixture of these two patterns was recorded in intermediate areas (the outer circular muscle or OCM) while triangular slow waves were present near the the submucosal plexus (SMP) inner circular muscle. Neither type of slow waves was affected by atropine, guanethidine, propranolol, and phentolamine (all 1 μM). Under these conditions of inhibition of NANC (non-adrenergic, non-cholinergic) nerves, electrical field stimulation (EFS) produced a fast, monophasic inhibitory junction potential (IJP) followed by a triggered slow wave (TSW) which could be premature or delayed and whose amplitude was maximum near the MyP region and decayed progressively in the other areas (minimum in SMP region). The K+ channel blocker, apamin at 10⁻⁶ M, did not affect resting membrane potentials or spontaneous slow waves but inhibited the amplitude of the IJP up to 70% and slightly but significantly enhanced (30%) the amplitude of the TSW. Long duration, single pulses (50-100 msec square waves, 10-20 V) elicited TSWs without IJPs. Both the slow waves and TSWs were associated with contractions of circular muscle which were significantly enhanced by apamin but not by blockers of adrenergic and cholinergic nerves. When the IJPs recorded near the MyP or DMP were abolished by tetrodotoxin (TTX, 1 μM) or by the NO synthase (NOS) inhibitor, N^ω nitro L-arginine (L-NNA, 50 μM), the occurrence of the TSW in response to EFS was advanced in time and increased in amplitude. The effects of L-NNA were reversed by L-but not D-arginine (both 1 mM). L-arginine significantly prolonged the durations of IJPs from the MyP and DMP regions. In contrast, the N-type Ca²⁺ channel blocker ω-conotoxin GVIA (ω-CTX, 1-3 x10⁻⁷M) abolished the IJP but delayed the induction of the TSW. Subsequent addition of either TTX or L-NNA advanced the onset of the TSW. The TSWs elicited by 50-100 msec single pulses were resistant to TTX, ω-CTX, or L-NNA. All treatments which abolished the IJP significantly increased contractions of circular muscle associated with spontaneous slow waves and TSWs. In the isolated circular muscle preparation (with the DMP intact) triangular slow waves were recorded near the DMP or close to the MyP border. The frequency and amplitude of the slow waves recorded near the DMP were significantly smaller than those recorded in similar areas in the full thickness preparation. EFS of this preparation evoked IJPs of 18-20 mV in amplitude. The IJPs were biphasic, lasted 5s and showed a fast and a slow component. No TSW occurred after the fast component of the IJPs; slow repolarization was observed instead. Long duration single pulses did not induce TSWs. In this preparation, the NOS inhibitor, N^ω nitro L-arginine methyl ester (L-NAME, 3x10⁻⁴ M), abolished the IJPs and regularized the slow waves, but TSWs could not be evoked. Superfusion of inhibitory neuromediators had different effects on pacemaking activity. SIN-1, a donor of NO, hyperpolarized the membrane, significantly increased slow wave frequency but reduced its amplitude, and abolished contractions. VIP (less effective) and PACAP (more effective) reduced slow wave frequency and amplitude without changing resting membrane potentials. P ACAP, but not VIP, increased circular muscle tone at 10⁻⁶ M. Nifedipine (10⁻⁷ and 3 X 10⁻⁷ M), an L-type calcium channel blocker, did not modify the shape of slow waves in any area of the full thickness preparation. It also did not reduce the amplitude of the IJP or TSW. Ni²⁺ at 200 μM, a Ca²⁺ channel blocker, inhibited slow wave frequency and amplitude and contractions. In Ca²⁺ -free Krebs (0.1 mM EGTA) for 10-15 min, the amplitude and frequency of the slow waves were gradually reduced. The TSW in response to 100 msec single pulses was still recorded near the MyP but never near the DMP region. The inhibitory effect of Ca²⁺ -free solution on slow wave amplitude was more rapid in onset near the DMP region. The intracellular Ca²⁺ store pump inhibitor, cyclopiazonic acid (10-30 μM), enhanced slow wave frequency and contractions. This differential sensitivity to removal of Ca²⁺ may be related to the morphological and functional observations which suggested that different electrical coupling properties between the pacemaker networks existed. The MyP pacemakers were less electrically well-coupled by visible gap junctions (low resistive cell-to-cell contacts) to outer circular muscle and hence showed greater susceptibility to 1 mM octanol (a gap junction blocker). The DMP pacemakers made numerous gap junction contacts to circular muscle, and slow waves paced from this region were less susceptible to 1 mM octanol. We conclude that 1) the pacemaker system of the canine ileum consists of two types of pacemakers that correspond to the presence of two networks of pacemaker cells found in the MyP and the DMP. The MyP network appeared to dominate pacemaking activity. 2) The slow waves and the TSW originated independently of neural activity but were delayed by IJPs. The MyP and the DMP provide two independent inhibitory neural inputs, where NO is released to mediate IJPs and relaxation and influence the delay in the occurrence of the TSW. 3) The TSW originates exclusively from the MyP region from which it spreads passively to other areas. It can reset the timing of slow waves in both pacemaker networks. 4) Ca²⁺ entry through non L-or N-type Ca²⁺ channels initiates slow waves. Intracellular Ca²⁺ stores modulate slow waves. 5) Different nature of electrical coupling of the MyP and DMP pacemakers to circular muscle may explain the differential sensitivity of slow waves to Ca²⁺ removal and gap junction blockade. / Thesis / Master of Engineering (ME)

The aim of the study was to observe changes in illeal digestibility of calcium and phosphorus during shell formation. The experiment was conducted on 350 Lohmann LSL-Classic hybrids laying whiteshelled eggs. At the time of the experiment, layers were fed with 115 g per layer per day of DeHeus N1 Power with the supplement of 0,5 % of chromium dioxide. Five groups were established. T1 (5–6 hours after laying), T2 (7–8 hours after laying), T3 (9–10 hours after laying), T4 (11–12 hours after laying), T5 (13–14 hours after laying). Six replications for each group were performed. All laying hens were housed in the same conditions in enriched cages. The ileal digestibility of calcium and phosphorus was determined by the indicator method. Within the first 10 hours after laying the last egg, the digestibility of calcium stayed unchanged at 61–62 %. A statistically significant increase in digestibility up to 75 % occurred approximately 12 hours after the lay in the T4 group. The digestibility of the phosphorus was fluctuating across all groups. Extension of the study throughout the whole process of eggshell formation would show a more comprehensive view of the issue.

In order to characterize peptide transport in the ovine small intestine, [14C]-glycyl-sarcosine uptake by tissue collected from five sheep was studied through the use of brush border membrane vesicles (BBMV). Preliminary experiments determined that incubation in hyaluronidase is not necessary in order to separate mucosal tissue from the basement membrane and that the stop solution used in the uptake study needed to be buffered. Uptake was examined in proximal (denoted jejunal) and distal (denoted ileal) halves of the intestine at four times (15, 30, 45, and 60 s) and at three extravesicular pH levels (6.4, 7.0, and 7.5). An intravesicular pH of 7.5 was used throughout the study. The two tissue sites differed (P < .02), with BBMV from jejunal tissue showing greater uptake than ileal. Uptake plateaued after 45 s, resulting in a quadratic (P < .005) effect of time. The effect of changes in extravesicular pH was also quadratic (P < .04), with uptake being greatest at pH 6.4, lowest at pH 7.0 and intermediate between the two at pH 7.5. Peptide uptake by sheep jejunal and ileal BBMV was demonstrated, but there was no clear evidence for increased uptake with decreasing extravesicular pH. / Master of Science

Salvia miltiorrhiza (SM) preconditioning was reported to be helpful in the early recovery of gastrointestinal motility in the intestinal congestion of rats with hepatic ischemia reperfusion. The aim of this study was to determine whether SM stimulates contraction of isolated terminal ileum of Sprague-Dawley rat ex vitro and the mechanisms which regulates that. The roots of SM were extracted by ethanol. One of the indicative marker of SM, Tanshinone IIA, was identified and quantified with high performance liquid chromatography (HPLC), and the results showed that Tanshinone IIA was 1190 £gg/ml in SM extract. The effects of contractile activity of SM extract at various cumulative dosages on the rat isolated terminal ileum were studied in organ bath. The area under curve above the baseline of contractile graphy of SM extract on isolated terminal ileum was recorded. In order to explore the contractile mechanism of SM extract on isolated terminal ileum, the individual pretreatment or use of atropine (a muscarinic receptor antagonist), tetrodotoxin (a sodium channel blocker), nifedipine (a calcium channel blocker), Ca2+ free Kreb¡¦s solution with EGTA, or trifluoperazine (a calmodulin blocker) was given and then cumulative dosages (40 £gL, 100 £gL, 180 £gL, 280 £gL) of SM extract were added. In addition, we used Fura-2 pentakis acetoxymethyl ester to detect the change of intracellular calcium concentration of intestinal epithelial cell-6 (IEC-6) induced in 1/1000-time or 1/10000-time dilution of SM extract. The result indicated SM extract significantly simulated the contraction of isolated terminal ileum in a dose-dependent manner. The individual addition or use of atropine, tetrodotoxin, nifedipine, or Ca2+ free Kreb¡¦s solution with EGTA all could not down-regulate significantly the contraction of SM extract on isolated terminal ileum. Trifluoperazine significantly down-regulated the contraction of SM extract on isolated terminal ileum. In addiation, SM extract was able to increase cytosolic calcium concentration of IEC-6 cells. In conclusion, the mechanisms of contraction of SM extract on isolated terminal ileum of rat were involved in calmodulin/Ca2+ associated contraction pathway.

Electrolyte transport in the intestine of the common brushtail possum (Trichosurus vulpecula) differs from that observed in eutherian mammals. This study has used molecular physiology to identify and characterise the expression and distribution of membrane transporters potentially responsible for these differences in electrolyte transport in the possum intestine. In the possum ileum, secretagogues stimulate an electrogenic Cl⁻-independent HCO₃⁻ secretory response but secretagogue-stimulated Cl⁻ secretion does not occur in this tissue. Based on the ion dependence and pharmacology of the stimulated secretory response, the expression of the cystic fibrosis transmembrane conductance regulator (CFTR), pancreatic Na⁺ HCO₃⁻ cotransporter (pNBC) and Na⁺ K⁺ 2Cl⁻ cotransporter (NKCC1) were investigated in the ileum. Reverse transcription PCR experiments showed that CFTR, pNBC and NKCC1 mRNA transcripts were expressed in the ileal epithelium. It was then demonstrated by in situ hybridisation that both CFTR and pNBC were localised predominantly in the crypts and the levels of expression decreased along the crypt-villous axis towards the lumen. Significantly, the in situ hybridisation results showed that there were low levels of NKCC1 transcript in the ileal epithelium. Western blot studies confirmed that mature CFTR and pNBC proteins were expressed in the ileum, while NKCC1 protein was not detected. The findings of the present study suggest that the absence of Cl⁻ secretion in the ileum is because NKCC1 expression is not elevated in the epithelium. The expression of mature CFTR and pNBC protein suggest that these membrane transporters are involved in the stimulated electrogenic HCO₃⁻ secretory response. The evidence also suggests that CFTR may mediate HCO₃⁻ efflux in the ileum. In contrast, secretagogues do not stimulate an electrogenic secretory response in the proximal and distal colon. This study has shown that CFTR, NKCC1 and pNBC proteins are expressed in the proximal and distal colon. Both NKCC1 and pNBC transcripts were localised to the crypt base in the proximal colon. However, it was shown that CFTR has a punctate distribution and the transcript was predominantly observed in the upper crypt and surface cell region. This study indicated that NKCC1 and pNBC were distributed in a different region of the epithelium compared to CFTR. It was concluded that the distribution of these membrane transporters in different regions of the epithelium accounts for the absence of a stimulated electrogenic secretory response in the possum colon. Given that no stimulated electrogenic secretory response is observed in the colon, it is suggested that HCO₃⁻ secretion by the ileum may have an important physiological role in maintaining an appropriate fluid and pH composition for fermentation in the colonic lumen.