Deirdre P. Campion

Eosinophil-nerve interactions and neuronal plasticity in rat gut associated lymphoid tissue (GALT) in response to enteric parasitism.

Intestinal lymphoid tissues and Peyers patches (PP) are innervated sites of immune surveillance in the gastrointestinal tract. Following infection with F. hepatica, neuronal hyperplasia and significantly increased eosinophil and mast cell trafficking to colonic PP sites were evident in rat tissues. Nerve-eosinophil associations were significantly elevated in infected colon and colonic PP, as were colonic tissue levels of the circulatory recruitment factors IL-5 and eotaxin. Increased immunoreactivity for neuronal plasticity markers GAP-43 and neural cell adhesion molecule (NCAM) was also found in infected tissues. Such neuronal alterations in the PP during enteric parasitism may have functional consequences on particular or pathogen uptake.

Reflex Cardiorespiratory Effects of Nociceptive Oesophageal Distension in the Decerebrate Rat

It is well established that painful distension of hollow viscera such as the oesophagus can evoke a reflex tachycardia and pressor response; however, the nature of the oesophageal afferent pathway(s) remains controversial. This study investigated the afferent arc which mediates these reflex cardiovascular changes in the decerebrate rat. In addition, the effect of oesophageal distension on the respiratory activity of the costal diaphragm was studied. Focal distension of the oesophagus (volume of 0.3 ml applied for 10 s) just above the diaphragmatic hiatus evoked a reproducible presssor response and tachycardia in the decerebrate rat. Respiration was transiently inhibited at the beginning of oesophageal distension and prior to the rise in blood pressure. Neuromuscular blockade with the nicotinic acetylcholine receptor blocker α‐bungarotoxin (140 μg bolus) had no effect on the magnitude of the cardiovascular response. Therefore the efferent supply to the striated muscle of the rat oesophagus was not essential in mediating this reflex. Signal averaging of the mean blood pressure response showed that neither selective ablation of oesophageal spinal afferents nor bilateral vagotomy altered the early trajectory of the pressure response. Bilateral vagotomy reduced the peak magnitude of the response to sustained oesophageal distension. In contrast, selective removal of spinal afferents had no effect on the response. Ablation of both neural pathways was essential to abolish the reflex cardiovascular and respiratory responses. It can be concluded that both vagal and spinal afferent pathways are utilised in the reflex cardiorespiratory response to painful oesophageal distension. Although ablation of one neural pathway had no effect on the response it was still implicated in the reflex, since ablation of both pathways was necessary to prevent the cardiorespiratory changes. This study emphasises the need for caution when inferences are made concerning single selective ablations of multiply innervated organs.

Chloride-led Disruption of the Intestinal Mucous Layer Impedes Salmonella Invasion: Evidence for an 'Enteric Tear' Mechanism

Background/Aims: Methods: The intestinal epithelial layer can switch from a net absorptive state to one of net secretion in the presence of luminal toxins and pathogens. This suggests an innate defence role for regulated secretion of mucus, electrolytes and water. We hypothesised that chloride-led fluid secretion across the mucus-covered intestinal epithelium alters barrier properties by influencing the overlying mucous-gel layer. Results: We demonstrated that chloride-led disruption of the epithelial-associated mucus-gel covering HT29-MTX-E12 (E12) human colonic epithelial monolayers, a goblet-cell like line derived from parental HT29 cells, resulted in reduction of associated mucus as well as a reduction in mucous-gel density and barrier properties. Changes in epithelial secretory state were accompanied by increased water transport, and the resulting loss of gel integrity reduced Salmonella typhimuriuminvasion of epithelia in both E12 monolayers and of isolated rat ileal mucosae. However, neither chloride secretion nor mucus disruption altered numbers of adhering bacteria. Conclusion: These data suggest a role for chloride led fluid secretion in the shedding of the adherent mucous-gel layer, possibly as a rate-limiting innate defence mechanism, and offer evidence for “enteric tears” in intestinal host defence.

A gastrointestinal role for the amphibian ‘diaphragm’ of Xenopus laevis

The ‘diaphragm’ of Xenopus laevis has close anatomical relations to the lower end of the oesophagus. In mammals, the crural diaphragm acts as a pinch valve at the gastro-oesophageal junction and is an important component of the gastro-oesophageal reflux barrier. The present study analysed the effect of amphibian ‘diaphragm’ contraction on oesophageal pressure using a superfused in situ oesophago-diaphragmatic preparation of large female Xenopus. Three-dimensional reconstruction of the oesophageal pressure profile was performed using four-port oesophageal infusion manometry. Bilateral electrical stimulation of the nerves supplying the ‘diaphragm’ of Xenopus increased the pressure volume vector of 5 mm of oesophagus (centred around the insertions of the diaphragm) from 20.4 ± 16 to 553.6 ± 232 mm · mmHg2 (mean ± SD). This was a statistically significant increase and statistically significantly higher than that evoked by electrical stimulation of both vagi (28.1 ± 30.7 mm·mmHg2). The amphibian ‘diaphragm’ seems to be functionally similar to the mammalian crural diaphragm. By analogy, we suggest that the original role of the diaphragm was not respiratory but gastrointestinal.

Effect of timing of post-partum introduction to pasture and supplementation with Saccharomyces cerevisiae on milk production, metabolic status, energy balance and some reproductive parameters in early lactation dairy cows.

Dietary change, an inconsistent nutrient intake and high levels of milk production make the early post-partum period (PP) a challenging time for the lactating dairy cow. This experiment investigates the effects of two early PP nutritional management strategies (NM): abrupt introduction to pasture (AP) or a total mixed ration (TMR) for 21 days followed by a gradual introduction to pasture over 7 days (GP), with (Y) or without (C) live yeast (YS) on milk production, energy balance (EB) and selected metabolic and reproductive variables. Forty multiparous dairy cows were assigned to one of four dietary treatments in a two (AP vs. GP) by two (Y vs. C) factorial, randomized block design. The experiment was conducted from days 1 to 70 PP. Blood samples were taken on day 1, day 5 and every 10 days until day 45 to determine metabolites, whilst intake (DMI), and EB were determined during week 6 PP. Milk was sampled weekly for fat, protein and lactose. Trans-rectal scanning for reproductive variables commenced on day 10 PP. Animals in the GP group had a higher DMI (p = 0.04), higher fat yield (p = 0.08) and fewer days to first ovulation (p = 0.09) vs. those in the AP group. EB (-3.5 ± 0.67 units of energy for milk production) and body condition score loss (0.70 ± 0.09) were not affected by NM. However, non-esterified fatty acids (NEFA) (p < 0.01) were higher, and glucose (p = 0.02) was lower in the AP vs. the GP group. Supplementary YS tended to improve EB (p = 0.09) and reduced NEFA (p < 0.01) vs. non-supplemented animals. These data suggest that offering animals a nutritionally balanced TMR during the first 3 weeks PP followed by a gradual introduction to pasture can improve DMI vs. pasture-based diets. Additionally, the blood metabolic profile suggests a more favourable energy status in the GP group or where YS was supplemented during the early PP period.

A comparison of the inhibitory effects of bupivacaine and levobupivacaine on isolated human pregnant myometrium contractility.

BACKGROUND: Epidural analgesia with levobupivacaine and bupivacaine is a common and effective method of labor pain relief. However, its use is associated with an increased instrumental delivery rate. One of the mechanisms postulated to account for this unwanted effect is the direct effect of local anesthetics on myometrial contractility. We determined the effects of bupivacaine and levobupivacaine on the amplitude and frequency of contractions of human term myometrium. METHODS: Uterine specimens were obtained from nonlaboring parturients scheduled for elective lower-segment cesarean delivery at term. Longitudinal muscle strips were prepared and mounted vertically in tissue chambers, and changes in the amplitude (peak force) and the frequency of contractions were recorded. Spontaneous contractions commenced after a period of application of 1 g (9.81 mN) of tension to the myometrial strips. No uterotonic drugs were used. The muscle strips were then exposed to cumulative concentrations of bupivacaine and levobupivacaine and dose–response curves were generated. RESULTS: Both bupivacaine and levobupivacaine decreased the amplitude of contractions in human myometrium in a concentration-dependent manner, reaching significance at 1 × 10−4 M for both bupivacaine and levobupivacaine compared with the internal control amplitude. With both drugs, the decrease in amplitude was accompanied by an increase in the frequency of contractions reaching significance at 3 × 10−5 M for both bupivacaine and levobupivacaine compared with the internal control frequency. CONCLUSIONS: The concentrations required for the effects on amplitude are much higher (33 fold) than the clinically relevant plasma concentrations of these drugs after epidural administration, and are unlikely to be significant in the setting of low-dose epidural analgesia in labor.

Opioid mediated activity and expression of mu and delta opioid receptors in isolated human term non-labouring myometrium

The existence of opioid receptors in mammalian myometrial tissue is now widely accepted. Previously enkephalin degrading enzymes have been shown to be elevated in pregnant rat uterus and a met-enkephalin analogue has been shown to alter spontaneous contractility of rat myometrium. Here we have undertaken studies to determine the effects of met-enkephalin on in vitro human myometrial contractility and investigate the expression of opioid receptors in pregnant myometrium. Myometrial biopsies were taken from women undergoing elective caesarean delivery at term. Organ bath experiments were used to investigate the effect of the met-enkephalin analogue [d-Ala 2, d-met 5] enkephalin (DAMEA) on spontaneous contractility. A confocal immunofluorescent technique and real time PCR were used to determine the expression of protein and mRNA, respectively for two opioid receptor subtypes, mu and delta. DAMEA had a concentration dependent inhibitory effect on contractile activity (1 × 10(-7)M-1 × 10(-4)M; 54% reduction in contractile activity, P<0.001 at 1 × 10(-4)M concentration). Mu and delta opioid receptor protein sub-types and their respective mRNA were identified in all tissues sampled. This is the first report of opioid receptor expression and of an opioid mediated uterorelaxant action in term human non-labouring myometrium in vitro.

Peripheral and gastrointestinal immune systems of healthy cattle raised outdoors at pasture or indoors on a concentrate-based ration

BackgroundDespite an increasing preference of consumers for beef produced from more extensive pasture-based production systems and potential human health benefits from the consumption of such beef, data regarding the health status of animals raised on pasture are limited. The objective of this study was to characterise specific aspects of the bovine peripheral and the gastrointestinal muscosal immune systems of cattle raised on an outdoor pasture system in comparison to animals raised on a conventional intensive indoor concentrate-based system.ResultsA number of in vitro functional tests of immune cells suggested subtle differences between the animals on the outdoor versus indoor production systems. There was a decrease in the number of neutrophils and monocytes engaged in phagocytosis in outdoor cattle (P < 0.01 and P < 0.05, respectively) in comparison to those indoors. Following mitogen stimulation, a lower level of interferon-γ was produced in leukocytes from the outdoor animals (P < 0.05). There was evidence of a gastrointestinal nematode infection in the outdoor animals with elevated levels of serum pepsinogen (P < 0.001), a higher number of eosinophils (P < 0.05) and a higher level of interleukin-4 and stem cell factor mRNA expression (P < 0.05) in the outdoor animals in comparison to the indoor animals. Lower levels of copper and iodine were measured in the outdoor animals in comparison to indoor animals (P < 0.001).ConclusionDespite distinctly contrasting production systems, only subtle differences were identified in the peripheral immune parameters measured between cattle raised at pasture in comparison to animals raised on a conventional intensive indoor concentrate-based production system.

Oral delivery of pathogens from the intestine to the nervous system.

Most therapeutic agents are delivered orally. Consequently, the major classes of therapeutically useful chemicals are partially lipophilic, small molecular weight compounds. They have reasonable permeability coefficient values across cell membranes, including those of intestinal epithelia and vascular endothelia. In contrast, large molecular weight biotechnology compounds have limited usefulness by non-injected routes as a consequence of their low membrane permeability and variable solubility. However, a wide range of infectious agents have developed strategies or have hijacked physiological routings in order to enter the host by the oral route. Efforts to address such issues have refreshed interest in mechanisms by which different types of payloads (including particulates and microorganisms) translocate across gut epithelia and then distribute to target tissues. Special attention is given to the potential role of the enteric nervous system and its plasticity.

An Investigation into the Mechanism of Action of Almitrine on Isolated Rat Diaphragm Muscle Fatigue

Background: Previous studies have shown that almitrine bismesylate, a respiratory stimulant which acts on the mitochondrial electron transport chain, enhances recovery of rat diaphragm muscle from fatigue. Objectives: Our aim is to investigate if the enhanced recovery is due to an anti-oxidant property of almitrine, since the electron transport chain is a major site of intracellular free radical production. Methods: A low-frequency fatigue protocol was used (30 Hz; 250 ms; delivered once every 2 s for 5 min), and the effects of almitrine before and after fatigue onset were compared to those of the anti-oxidant compound N-acetylcysteine (NAC). Results: Almitrine (6 and 10 µg/ml) given before fatigue gave better recovery rates than postfatigue application. In contrast, NAC (100 µM) application before fatigue onset was not as effective as NAC given immediately after the cessation of the fatigue protocol. However, almitrine (6 µg/ml) completely reversed the reduction in baseline twitch tension brought about by a free-radical-producing mixture of FeCl3 + ADP (1 mM + 2.5 mM, respectively). Conclusion: The results of this study confirm that almitrine enhances recovery from fatigue and, in contrast to NAC prefatigue application, is more effective. Also, almitrine was shown to have an anti-oxidant effect, but it does not act like a typical anti-oxidant.