Effects of the probiotic yeast Saccharomyces boulardii on the neurochemistry of myenteric neurones in pig jejunum

We studied the effects of food supplementation with Saccharomyces boulardii (S. boulardii; synonym S. cerevisiae HANSEN CBS 5926; 1 g per day for 9 days) on the presence and co-localization patterns of neuronal markers in myenteric neurones of the pig jejunum. The pan neuronal marker Hu revealed no change in the number of neuronal cell bodies per ganglion (37 +/- 7 in control vs 34 +/- 9 in the S. boulardii group). Ranked by size the following cell populations were identified: choline acetyltransferase (ChAT), calbindin-28k (CALB), substance P (SP), neurofilament 160 kD (NF-160), vasoactive intestinal polypeptide (VIP), nitric oxide synthase (NOS), calcitonin gene-related peptide (CGRP), calretinin (CALRET). We found a significant decrease in the number of CALB myenteric neurones in animals which received S. boulardii supplemented diet. None of the other neuronal markers revealed any difference between controls and S. boulardii treated animals. The study reports transmitter-localization patterns in the myenteric plexus of the pig jejunum and provides evidence that changes in the neurochemistry of enteric neurones occur with S. boulardii supplemented diet. Although only CALB expression was altered and the functional significance of this finding remains unknown, our study identified a possible new effector level of probiotics in the gut.     

Preventive Effects of the Probiotic Escherichia coli Strain Nissle 1917 on Acute Secretory Diarrhea in a Pig Model of Intestinal Infection

Pretreatment with the probiotic Escherichia colistrain Nissle 1917 (EcN) was assessed in a pig model of intestinal infection to prevent acute secretory diarrhea. In the model 10¹⁰ colony forming units of the porcine enterotoxigenic Escherichia coli Abbotstown (EcA) was given via orogastric tube to weaned piglets at day 21 postpartum (−EcN/+EcA group, n = 7). Forty-eight hours after challenge electrophysiological parameters of isolated intact jejunal epithelia were characterized in Ussing chambers. In agreement with clinical signs of diarrhea, tissues of challenged animals showed an overshoot of secretory response after stimulation of the cAMP-mediated second messenger pathway by forskolin, indicating higher excitability of chloride secretory systems under infected conditions. The data were compared with respective measurements from animals that got a daily dose of 10¹⁰ cfu of the probiotic EcN over 10 days before EcA challenge (+EcN/+EcA group; n = 4), from a group that received only EcN (+EcN/–EcA; n = 4), or from a group that remained totally untreated (−EcN/−EcA; n = 6). EcN pretreatment completely abolished clinical signs of secretory diarrhea in +EcN/+EcA animals. Furthermore, jejunum epithelia of these animals did not exhibit an overshoot of secretory response upon stimulation with forskolin. Our studies demonstrate for the first time the efficacy of prophylactic EcN in pig small intestine for preventing an effect of toxigenic EcA. This infection model with freshly weaned piglets may be predestinated to further characterize EcN effects on the cellular level, i.e., involved second messenger pathways, or it may also be useful to examine the efficacy of other substrates or microbe strains against secretory stimuli.     

Impaired Ca2+-handling in HIF-1α+/− mice as a consequence of pressure overload

The hypoxia-inducible factor (HIF)-1 is critically involved in the cellular adaptation to a decrease in oxygen availability. The influence of HIF-1α for the development of cardiac hypertrophy and cardiac function that occurs in response to sustained pressure overload has been mainly attributed to a challenged cardiac angiogenesis and cardiac hypertrophy up to now. Hif-1α ^+/+ and Hif-1α ^+/? mice were studied regarding left ventricular hypertrophy and cardiac function after being subjected to transverse aortic constriction (TAC). After TAC, both Hif-1α ^+/+ and Hif-1α ^+/? mice developed left ventricular hypertrophy with increased posterior wall thickness, septum thickness and increased left ventricular weight to a similar extent. No significant difference in cardiac vessel density was observed between Hif-1α ^+/+ and Hif-1α ^+/? mice. However, only the Hif-1α ^+/? mice developed severe heart failure as revealed by a significantly reduced fractional shortening mostly due to increased end-systolic left ventricular diameter. On the single cell level this correlated with reduced myocyte shortenings, decreased intracellular Ca²⁺-transients and SR-Ca²⁺ content in myocytes of Hif-1a ^+/? mice. Thus, HIF-1α can be critically involved in the preservation of cardiac function after chronic pressure overload without affecting cardiac hypertrophy. This effect is mediated via HIF-dependent modulation of cardiac calcium handling and contractility.     

Modulation of electrolyte homeostasis by dietary nitrogen intake in growing goats

In goats, the combination of dietary N and Ca reduction caused hypocalcaemia and further changes in Ca homeostasis. The aim of the present study was to characterise the effects of dietary N reduction under normocalcaemia on mineral and bone metabolism in young goats. Young male goats of the Saanen breed were fed a diet reduced in N (8 %) for about 7 weeks (ten animals per group) and were compared with goats fed with an adequate N (14 %) diet. When N intake was reduced in young goats, plasma urea concentrations as well as renal elimination of urea were reduced. This was inversely related to creatinine in plasma and urine, which increased during a dietary N reduction as a function of reduced renal activity to save urea during N scarcity. During this decrease in renal function, associated with declined insulin-like growth factor 1 concentrations, a reduction in calcidiol and calcitriol concentrations could be observed. Meanwhile, carboxyterminal cross-linked telopeptide of type I collagen values and activity of total alkaline phosphatase were both elevated, indicating some bone remodelling processes taking place during a reduced N diet in young goats. The concentrations of inorganic phosphate (Pi) and total Ca were changed in several body fluids, indicating that Pi and Ca homeostasis was perturbed in goats fed a reduced N diet. Therefore, more research is needed to find the balance between reduction of environmental N pollution by reducing dietary N in ruminant feeding and maintaining the animal's health.     

Immunopathogenesis of Ascaridia galli infection in layer chicken

Gastro-intestinal nematode infections in mammals are associated with local T lymphocyte infiltrations, Th2 cytokine induction, and alterations in epithelial cell secretion and absorption. This study demonstrates that Ascaridia (A.) galli infection in chicken also elicits local gut-associated immune reactions and changes in the intestinal electrogenic nutrient transport. In A. galli-infected birds we observed infiltrations of different T cell populations in the intestinal lamina propria and accumulation of CD4+ lymphocytes in the epithelium. The Th2 cytokines IL-4 and IL-13 dominated the intestinal immune reactions following A. galli infection. A. galli-specific systemic IgY antibodies were detected after two weeks post infection, and did only poorly correlate with detected worm numbers. Electrogenic transport of alanin and glucose was impaired in A. galli-infected chicken. Our data provide circumstantial evidence that local immune responses and electro-physiological intestinal functions may be connected and contribute to the elimination of worm infection.     

Pre- and postprandial effects on ghrelin signaling in the brain and on the GH/IGF-I axis in the Mozambique tilapia (Oreochromis mossambicus)

The discovery of ghrelin (GRLN) has broadened our understanding of the regulation of energy homeostasis in vertebrates. In addition to stimulating growth hormone release from the pituitary, GRLN has been implicated as a hunger signal stimulating food intake in mammals and goldfish. Indeed, GRLN levels rise preprandial and fall following a meal. The current study investigated pre- and postprandial changes (3 h before and after a meal) in GRLN signaling in the tilapia (Oreochromis mossambicus). Significant elevations in preprandial brain mRNA levels of the GRLN receptor (GHS-R1a) and GRLN were observed; though not significant brain neuropeptide Y (NPY) mRNA levels did increase preprandially. GHS-R1b, and NPY mRNA levels were reduced significantly 3 h after a meal; whereas GHS-R1a levels were unaltered postprandially. Brain ghrelin mRNA levels exhibited a transient significant increase 1 h postprandially. Tilapia that missed the scheduled feeding exhibited no changes in brain GHS-R1a, GRLN and NPY postprandial mRNA levels; whereas GHS-R1b mRNA levels were significantly reduced 1 and 3 h postprandially. Brain GHSR preprocessed RNA (heteronuclear mRNA) levels were significantly elevated 3 h preprandially. GHS-R hnRNA levels were significantly elevated 1 h postprandial in fed and fasted tilapia. No preprandial rise in plasma GRLN was observed. Following a meal, plasma GRLN levels were significantly elevated; whereas there was no change in tilapia missing the scheduled feeding. Stomach mRNA levels of GRLN rose preprandially and remained unchanged following a meal. In animals that missed the scheduled feeding stomach GRLN levels dropped significantly 1 h following a meal. There was no change in plasma growth hormone levels in the fed fish, although there was a significant rise in the fasted fish 1 h after the scheduled feeding. Postprandial levels of plasma IGF-I were elevated in both fed and fasted tilapia. These results suggest that brain derived GRLN is likely driving day-to-day appetite through GHS-R1a and NPY; while systemic GRLN may play a role in postprandial metabolism.     

Switching of Na+, K+-ATPase isoforms by salinity and prolactin in the gill of a cichlid fish

We identified and investigated the changes in expression of two gill Na(+), K(+)-ATPase α-subunit isoforms (α-1a and α-1b) in relationship with salinity acclimation in a cichlid fish, Mozambique tilapia. Transfer of freshwater (FW)-acclimated fish to seawater (SW) resulted in a marked reduction in α-1a expression within 24?h and a significant increase in α-1b expression with maximum levels attained 7 days after the transfer. In contrast, transfer of SW-acclimated fish to FW induced a marked increase in α-1a expression within 2 days, while α-1b expression decreased significantly after 14 days. Hypophysectomy resulted in a virtual shutdown of α-1a mRNA expression in both FW- and SW-acclimated fish, whereas no significant effect was observed in α-1b expression. Replacement therapy by ovine prolactin (oPrl) fully restored α-1a expression in FW-acclimated fish, while cortisol had a modest, but significant, stimulatory effect on α-1a expression. In hypophysectomized fish in SW, replacement therapy with oPrl alone or in combination with cortisol resulted in a marked increase in α-1a mRNA to levels far exceeding those observed in sham-operated fish. Expression of α-1b mRNA was unaffected by hormone treatment either in FW-acclimated fish or in SW-acclimated fish. The mRNA expression of fxyd-11, a regulatory Na(+), K(+)-ATPase subunit, was transiently enhanced during both FW and SW acclimation. In hypophysectomized fish in FW, oPrl and cortisol stimulated fxyd-11 expression in a synergistic manner. The clear Prl dependence of gill α-1a expression may partially explain the importance of this hormone to hyperosmoregulation in this species.     

Colonic Absorption and Bioavailability of the Pentapeptide Metkephamid in the Rat

The concept of delivering systemically active peptide drugs to the colon in order to improve their oral absorption requires reasonable peptide permeability of the large intestinal wall and stability against the activity of the colonic microflora. In addition, the role of hepatic extraction needs to be addressed. In this study the absorption of the pentapeptide metkephamid following single pass perfusion of rat ascending colon was investigated by monitoring its disappearance from the large intestine and simultaneous appearance in the portal vein, the hepatic vein and the aorta. In addition its stability against colonic microflora was tested in vitro using pig caecal contents. Metkephamid was absorbed from the large intestine and appeared in the blood circulation; peptide concentrations in the portal vein increased over-proportionally with increasing perfusate concentrations (0.1 – 4.6 mmol/L) from 0.19 µg/mL ± 0.12 (SD, n = 7) to 31.6 µg/mL + 20.65 (SD, n = 4), respectively, and thus suggesting a saturable transport or metabolism. Concentrations in the hepatic vein were significantly lower than in the portal vein, hepatic extraction ratios were 0.35 ± 0.14, 0.61 ± 0.18 and 0.62 ± 0.28 (SD, n = 4) for 0.1, 0.5 and 1.0 mM metkephamid perfusate concentrations, respectively. In the anaerobic colon metabolism model the degradation half-life of the peptide was 14.9 hours, thus, indicating relative stability in the bacterial environment of the colon. The results of the present study encourage further investigations on colonic delivery of peptide drugs.