Biomedical subjects

N M Griffiths

Publications and source records attributed to N M Griffiths.

At least 37 records · Page 2Linked to original sources

PubMed2000

Functional changes in the rat distal colon after whole-body irradiation: dose-response and temporal relationships.

The aim of this study was to determine the acute radiation response of the rat distal colon by in vivo and in vitro measurements of the functions of the colon over a range of radiation doses. Rats received a whole-body irradiation of 2 to 12 Gy and were studied from 1 to 7 days after exposure. In vivo water and electrolyte absorption was measured by insertion of an agarose cylinder in the colon of anesthetized rats. In vitro transepithelial electrical parameters (potential difference, short-circuit current, transepithelial conductance) were measured in Ussing chambers in basal and agonist-stimulated conditions. In vivo and in vitro functional studies were completed by standard histological analyses. The majority of functional modifications appeared at 4 days after exposure. At this time, a dose-dependent decrease in absorption of water and sodium/chloride ions in the colon was noted. In contrast, a twofold increase in potassium secretion was observed for every radiation dose studied. The response to secretagogues was attenuated at doses >8 Gy. Modifications of basal transepithelial electrical parameters together with marked histological alterations were observed at 4 days with the higher doses (>/=10 Gy). In conclusion, these results show that functions of the colon are affected by irradiation and may contribute to diarrhea induced by ionizing radiation.

Animals↗

PubMed1999

Long-term effects of X-irradiation on gastrointestinal function and regulatory peptides in monkeys.

PURPOSE: To investigate the long-term effects of X-irradiation on different aspects of gastrointestinal function in the non-human primate (Macaca mulatta). MATERIALS AND METHODS: Animals were exposed to X-radiation (5 or 6 Gy) or not (sham) and gastrointestinal function was investigated 4-6 years after exposure. Basal and agonist-stimulated short circuit current (Isc) responses were measured in isolated jejunum. Intestinal tissue was taken for histological analysis as well as for determination of mucosal marker enzyme activities and gastrointestinal regulatory peptide levels. Vasoactive intestinal peptide receptor characteristics were determined as well as VIP-stimulated Isc responses. GI peptides were also measured in plasma. RESULTS: Few differences were seen in basal electrical parameters or tissue morphology but there was a tendency for reduced basolateral membrane enzyme activity. VIP-stimulated Isc responses were reduced in irradiated animals as were VIP-stimulated adenylate cyclase responses. Plasma and tissue (ileal and colonic muscle layers) gastrin releasing peptide levels were increased in irradiated animals. In contrast circulating gastrin levels were lower. CONCLUSIONS: Late effects of total-body irradiation on GI function in monkeys showed altered circulating and tissue levels of some GI peptides. In addition the biological effects of vasoactive intestinal peptide were modified.

Adenylyl Cyclases↗

PubMed1999

Modified bile acid profiles in mixed neutron and gamma-irradiated pigs.

PURPOSE: To determine the effect of mixed neutron and gamma-irradiation on the bile acid pool, which may be a key factor in radiation-induced diarrhoea. MATERIALS AND METHODS: The bile duct of pigs was catheterized to derive bile over several experimental weeks, both before and after a 5.9 Gy neutron and gamma-irradiation. After measurement of the volume and sampling, bile was returned to the pig via a duodenal catheter. Samples of bile were analysed by HPLC for their individual and total bile acid content. Blood samples were also collected for total bile acid determination. RESULTS: Bile flow was significantly decreased during the first 24h and after the fifth day post-irradiation. Whereas total bile acid concentration in bile was not altered, profiles of individual bile acids were significantly altered as early as the first post-irradiation day. Such modifications in these profiles resulted in a change of the properties of the bile acid pool. An increased proportion of dihydroxylated bile acids known to be more deleterious for the intestine was observed. CONCLUSIONS: Neutron and gamma-irradiation leads to modifications of bile acid profiles, which may partly explain radiation-induced diarrhoea by a coherent physiopathological mechanism.

Animals↗

PubMed1999

Modified absorptive and secretory processes in the rat distal colon after neutron irradiation: in vivo and in vitro studies.

Impaired fluid and electrolyte transport in the intestine is a well-recognized characteristic of radiation-induced pathologies in the gastrointestinal tract. The aim of this study was to investigate the responsiveness of the epithelium of the colon of the rat to electrical and pharmacological (serotonin, carbachol) stimulation concomitantly with in vivo assessment of the absorptive capacity of the colon at 1, 3, 5 and 7 days after 3.8 Gy whole-body exposure to neutrons. The responsiveness of rat colon in vitro to electrical stimulation and the number of mast cells were measured to examine the role of neuroimmune networks in radiation-induced dysfunction. Animals showed an impaired capacity of the colon to absorb water and sodium from 3 to 5 days after irradiation together with decreased responsiveness to electrical and pharmacological stimulation. The time course of decreased responsiveness to neural stimulation was similar to that of impaired absorption observed in vivo, but it was not correlated with variations in mast cell numbers. Histological (mast cells) and biochemical analyses (myeloperoxidase and NO synthase activities) did not find evidence of a marked infiltration and/or activation of inflammatory cells. Thus the impaired absorptive capacity of the colon observed after irradiation occurs concomitantly with decreased neural influence, and is possibly related to reduced epithelial functional capacity but not to decreased mast cell numbers.

Animals↗

PubMed1998

In vivo alterations of fluid and electrolyte fluxes in rat colon by gamma irradiation.

Colonic function in rats was investigated up to 14 days following exposure to whole-body gamma irradiation (8 Gy) using a combination of in vivo and in vitro approaches. Water and electrolyte fluxes were measured in vivo under anesthesia by insertion of an agarose cylinder into the descending colon. Short-circuit current responses (Isc; basal, agonist-stimulated) of distal colon were measured in vitro as were mannitol and sodium fluxes. Water and electrolyte absorption (Na, Cl) was markedly reduced at four days after irradiation but returned to normal at seven days. Potassium secretion was increased from one to seven days after exposure. There were no differences in basal Isc, Na, or mannitol fluxes at four days but responses to secretagogues (5-hydroxytryptamine, forskolin, carbachol) were attenuated. No morphological alterations were associated with these functional modifications.

Animals↗

PubMed1998

A substance P receptor antagonist (FK 888) modifies gut alterations induced by ionizing radiation.

PURPOSE: We previously reported disturbances of ileal substance P (SP) levels and of characteristics of specific receptors after ionizing radiation associated with disorders of intestinal motility. The aim of this study was to investigate the effect of a SP receptor blockade by FK 888 on gut SP levels and contractile properties after rat irradiation. MATERIALS AND METHODS: Rats were exposed to 6 Gy whole-body gamma-irradiation and injected 1 h post-irradiation with FK 888 for 3 days (0.1 mg/kg/day). Plasma and ileal SP concentrations, ileal muscle SP receptor binding and SP-induced contractions in isolated ileum were investigated 3 and 14 days post-irradiation and FK 888 treatment. RESULTS: Irradiation induced an increase of total SP binding site number at day 3 (1.3-fold) and day 14 (1.6-fold). FK 888 had no effect on SP receptor characteristics in irradiated animals. In contrast, FK 888 treatment caused a reduction of endogenous ileal SP level in mucosal (-29%) and muscularis (-40%) layers at day 3 and these decreases were greater at day 14, -88% in mucosal and -61% in muscularis layers. FK 888 treatment decreased efficacy of ileal contraction in both the control and irradiated rat but surprisingly it increased potency at day 3 and decreased it at day 14 in the irradiated rat. CONCLUSIONS: The findings demonstrate that a SP receptor antagonist could be effective on intestine contractility alteration induced several days after ionizing radiation exposure but not at 3 days after irradiation.

Animals↗

PubMed1998

Exposure to ionizing radiation modifies circulating gastrin levels and gastrointestinal endocrine cell densities in the rat.

PURPOSE: Gastrointestinal functions, controlled partly by gut peptides, are disturbed by ionizing radiation exposure. The effect of whole-body irradiation on circulating gastrin levels, densities of gastrointestinal endocrine cells and gastric acid secretion was investigated. MATERIALS AND METHODS: Rats were exposed to 2 or 6 Gy gamma-radiation. They were killed 3 or 7 days later and compared with shams. Plasma gastrin and basal acid output were measured. Endocrine cells were identified by argyrophilia or immunohistochemistry and their densities estimated. RESULTS: Radiation exposure significantly increased gastrinaemia and gastric acid output at the times studied (p<0.05-p<0.001). Endocrine cells displayed different sensitivities to irradiation. In the gastric mucosa, a 6 Gy dose induced a decrease in fundic argyrophil cell, antral gastrin and somatostatin cell densities, always accentuated 7 days after irradiation, while in the intestinal mucosa it induced an increase, with highest values often at 7 days post-irradiation (p<0.01-p<0.001). This was true for neurotensin cells in the jejunum and ileum, substance P cells in ileum and enteroglucagon cells in the descending colon. CONCLUSIONS: Whole-body irradiation in rats significantly alters plasma gastrin levels, and several gut endocrine cell densities. This has repercussions on hormonal function, such as that exerted on acid secretion, and may explain gastrointestinal dysfunction observed following radiation exposure.

Animals↗

PubMed1998

Exposure to ionizing radiation modifies neurally-evoked electrolyte transport and some inflammatory responses in rat colon in vitro.

PURPOSE: To characterize the responsiveness of the colon to neural stimulation following acute exposure of rats to gamma-radiation and to correlate observed changes to a number of parameters. MATERIALS AND METHODS: Rats were exposed to 5 or 10 Gy 137Cs gamma-radiation or not (sham-irradiated) and studied at 1, 3 and 7 days after irradiation. Stripped segments of colon were mounted in Ussing chambers for measurements of neurally-evoked electrolyte transport (electrical field stimulation). Colonic tissue was also taken for biochemical (tissue 5-hydroxytryptamine, histamine, leukotriene B4, nitric oxide synthase) and histological analyses (mast cells). RESULTS: In irradiated rats both proximal and distal colon were hyporesponsive to electrical field stimulation at 1 and 3 days, but had recovered by 7 days. In the distal colon, carbachol responses were attenuated 1 day after 10 Gy. Mast cells, tissue histamine and leukotriene B4 synthesis were significantly reduced at all time points but no changes were seen in 5-HT or inducible NOS activity. CONCLUSIONS: Rat colon becomes hyporesponsive to neural stimuli post-irradiation. The response initially (1 and 3 days) correlates with decreased mast cells and histamine, but not at 7 days.

Animals↗

PubMed1998

Ionizing radiation stimulates muscarinic regulation of rat intestinal mucosal function.

The aim of this study was to determine whether ionizing radiation modifies muscarinic regulation of intestinal mucosal function. Rats exposed to total body 8-Gy gamma-irradiation or sham irradiated were studied up to 21 days after irradiation. Basal and carbachol-stimulated short-circuit current (Isc) and transepithelial conductance (Gt) of stripped ileum were determined in Ussing chambers. Muscarinic receptor characteristics using the muscarinic antagonist [3H]quinuclidinyl benzilate and three unlabeled antagonists were measured in small intestinal plasma membranes together with two marker enzyme activities (sucrase, Na+-K+-ATPase). Enzyme activities were decreased 4 days after irradiation (day 4). Basal electrical parameters were unchanged. Maximal carbachol-induced changes in Isc and Gt were increased at day 4 (maximal DeltaIsc = 195.8 +/- 14.7 microA/cm2, n = 19, vs. 115.4 +/- 8.2 microA/cm2, n = 63, for control rats) and unchanged at day 7. Dissociation constant was decreased at day 4 (0.73 +/- 0.29 nM, n = 10, vs. 2.14 +/- 0.39 nM, n = 13, for control rats) but unchanged at day 7, without change in binding site number. Thus total body irradiation induces a temporary stimulation of cholinergic regulation of mucosal intestinal function that may result in radiation-induced diarrhea.

Animals↗

PubMed1997

Changes in gut neurotensin and modified colonic motility following whole-body irradiation in rat.

Exposure to ionizing radiation induces gastrointestinal dysfunction often associated with disorders of intestinal motility. Neurotensin is one of the mediators involved in the control of intestinal muscle activity. The aim of this study was to relate neurotensin tissue content and specific receptor binding with contractile effect of neurotensin in rat colon after irradiation. Rats were exposed to whole-body gamma-irradiation (60Co; 6 Gy). Intestinal (caecum, colon) neurotensin-like immunoreactivity, colonic muscle neurotensin receptor binding and neurotensin-induced contractions in isolated colon were investigated 3 and 7 days after irradiation. Irradiation produced a marked increase in the intestinal muscle content of neurotensin-like immunoreactivity (2.5-fold in caecum, 5-fold in colon) 3 days post-irradiation. At 7 days, the intestinal neurotensin content was close to that of the control values. Three days after irradiation, neurotensin receptors in colonic muscle were characterized by the appearance of a transient second class of sites of low affinity-high capacity. A three-fold increase in the total number of sites was observed. In addition, effects of neurotensin on isolated colon preparations showed an increase (37%) of potency but a decrease (7-fold) of efficacy. Seven days after irradiation, the efficacy was close to the control. Modifications of intestinal neurotensin content and specific receptor characteristics induced by irradiation can influence the colonic contractile activity.

Animals↗

PubMed1997

Whole-body gamma irradiation modifies bile composition in the pig.

The effects of 6 Gy whole-body 60Co gamma irradiation on bile composition in pigs were studied to determine possible alterations in the quality of the bile, which may be a determining factor in diarrhea as well as nutrient malabsorption, which classically occurs after irradiation. The bile duct of pigs was catheterized to allow a total and continuous deviation of bile over several weeks, before and after irradiation. After measurement of the volume and sampling, bile was returned to the animal via a duodenal catheter. Bile samples were then analyzed for cholesterol, phospholipid and total bile acid content. Individual bile acids were quantified by HPLC analysis. Bile flow was significantly decreased during the first 24 h and after the fifth day postirradiation. Whereas cholesterol, phospholipid and total bile acid concentrations were not altered, profiles of individual bile acids were modified significantly as early as the first day postirradiation. Moreover, the change of these profiles with time was specific for each bile acid. Such modifications in bile acid profiles resulted in a change in the properties of the bile acid pool in an increased proportion of dihydroxylated bile acids known to interfere with gut functions, and it is reasonable to suggest that radiation-induced changes in bile acid profiles may be involved in radiation-induced gastrointestinal disorders.

Animals↗

PubMed1996

Exposure to either gamma or a mixed neutron/gamma field irradiation modifies vasoactive intestinal peptide receptor characteristics in membranes isolated from pig jejunum.

The effect of acute whole body exposure to ionizing radiation was investigated on intestinal vasoactive intestinal peptide (VIP) receptors and adenylate cyclase activity in membranes isolated from pig jejunum. Pigs under light anaesthesia were exposed to a single dose (6 Gy) of gamma (gamma) or to mixed neutron/gamma field (ratio 1:1; neutron/gamma) irradiation. Seven days after irradiation, plasma-membranes were prepared from post mortem jejunal mucosal scrapings. Marker enzyme activities (sucrase, leucine aminopeptidase (LAP), Na,K-ATPase) were measured in each preparation. The characteristics (KD, Bmax) of VIP receptors were determined using 125I-labelled VIP. In addition VIP-sensitive adenylate cyclase activity was measured. Results showed that enzyme activities were reduced following both gamma (sucrase 67%; LAP 53%; Na/K-ATPase 29%; N = 7) and neutron/gamma (sucrase 53%; LAP 59%; Na/K-ATPase 68%; N = 5) compared with control values (N = 5). VIP receptor affinity was decreased following either type of irradiation (gamma or neutron/gamma P < 0.01) and receptor numbers increased. Both VIP- and forskolin-stimulated adenylate cyclase activities were reduced but the sensitivity of the enzyme remained the same for VIP (EC50 values (nmol dm-3)-control-1.27 +/- 0.35; gamma-2.18 +/- 0.41; neutron/gamma-1.91 +/- 0.28). In conclusion, exposure to either gamma or neutron/gamma irradiation attenuates intestinal enzyme activities and VIP receptor affinity but increases VIP receptor numbers.

Adenylyl Cyclases↗

PubMed1994

Active intestinal secretion of the fluoroquinolone antibacterials ciprofloxacin, norfloxacin and pefloxacin; a common secretory pathway?

The transepithelial transport of three fluoroquinolones, norfloxacin, ciprofloxacin and pefloxacin has been compared by using cultured human intestinal Caco-2 cell-layers. Absorptive (apical-basal) fluxes of ciprofloxacin and norfloxacin are small relative to basal to apical fluxes. Norfloxacin and ciprofloxacin are thus subject to active transepithelial secretion. Active net secretion of norfloxacin displays saturation kinetics with Vmax and Km values of 36.2 +/- 6.9 nmol.cm-2.hr-1 and 1.42 +/- 0.79 mM. In contrast, transepithelial pefloxacin fluxes are large, show marked saturation while the direction of net flux is variable and small relative to transepithelial fluxes. Norfloxacin, pefloxacin and ciprofloxacin are all subject to accumulative transport across the basal surface of Caco-2 cell layers. A number of 4-quinolones and fluoroquinolones are capable of inhibition of both net secretion of ciprofloxacin and cellular accumulation across the basal-lateral cell surface. Cinoxacin, a 4-quinolone may selectively inhibit exit from the cell across the apical membrane. Cross-competition studies suggest that fluoroquinolones may compete for a common carrier at the basal-lateral membrane. It is likely that the mechanism of transepithelial secretion involves a common accumulative transport at the basal-lateral membrane followed by facilitated exist across the apical membrane. Pefloxacin may interact with a brush-border carrier for which norfloxacin and ciprofloxacin are poor substrates, enhancing the absorptive flux of this fluoroquinolone.

Biological Transport↗

PubMed1993

Effect of prostaglandin E2 on agonist-stimulated cAMP accumulation in the distal convoluted tubule isolated from the rabbit kidney.

The effects of calcitonin, vasoactive intestinal peptide (VIP), parathyroid hormone (PTH) and isoprenaline on intracellular cAMP accumulation were determined in the distal tubule (DCT) microdissected from collagenase-treated rabbit kidney. In DCTb (the initial "bright" portion) calcitonin (10 ng/ml) elicited a highly reproducible response 203.7 +/- 19.1 fmol cAMP mm-1 4 min-1 (SE,N = 13) whereas VIP-induced cAMP accumulation was less and more variable from one experiment to another (1 microM, 97.2 +/- 17.8 fmol mm-1 4 min-1, SE, N = 12). When used in combination, these two agonists were non-additive, indicating stimulation of a single pool of cAMP in DCTb. In DCTg, ("granular") which consists of at least two cell types, PTH (100 nM) elicited a marked, reproducible accumulation of cAMP (154.3 +/- 27.0 fmol mm-1 4 min-1; SE, N = 5). Isoprenaline (1 microM) and VIP (1 microM) induced much smaller increases in cAMP levels 20.9 +/- 2.7 and 29.4 +/- 4.1 fmol mm-1 4 min-1 (SE, N = 5) respectively, and, when used in combination, were non-additive, demonstrating that VIP and isoprenaline are active on the same cell type. In DCTb, prostaglandin E2 (PGE2) inhibited both calcitonin- and VIP-stimulated cAMP accumulation (calcitonin 57.8 +/- 2.7% inhibition, SE, N = 16; VIP, 80.6 +/- 2.1% inhibition, SE, N = 5). The EC50 values for calcitonin were 1.21 +/- 0.33 ng/ml and 1.83 +/- 0.25 ng/ml (SD, N = 3) in the absence and presence of PGE2 (300 nM) respectively with an IC50 for PGE2 of 26.3 +/- 6.3 nM (SE, N = 4).(ABSTRACT TRUNCATED AT 250 WORDS)

1-Methyl-3-isobutylxanthine↗

PubMed1993

Active secretion of the fluoroquinolone ciprofloxacin by human intestinal epithelial Caco-2 cell layers.

The bidirectional transepithelial fluxes of ciprofloxacin, an antibacterial fluoroquinolone, across the human intestinal epithelial Caco-2 cell-line show marked asymmetry. Basal-to-apical flux of ciprofloxacin (10 microM) exceeds apical-to-basal flux indicating net secretion. Net ciprofloxacin secretion is abolished by azide/2-deoxy-D-glucose treatment, displays saturation kinetics (Km = 0.89 +/- 0.23 mM, Vmax 44.3 +/- 4.9 nmol cm-2.h) and competition by other fluoroquinolones. A specific, active secretion in Caco-2 epithelia may explain the transintestinal elimination of ciprofloxacin observed in pharmacokinetic studies in man.

Azides↗

PubMed1991

Discrepancy between the short and long term effects of ouabain on the sodium pumps of human cells grown in culture.

1. Human cells (HeLa) were cultured for periods up to 48 h in growth medium in the absence or presence of a range of concentrations of cardiac glycosides. In some experiments the potassium concentration of the medium was varied between 0.3 mM and the usual 5 mM. 2. For periods up to 2 h in ouabain the association and dissociation rate constants were measured and the equilibrium binding constant (KD) calculated; the apparent equilibrium binding constant (K'D) was measured after 1-2 days growth in ouabain. 3. Ouabain had a K'D after 2 days of 2-6 nM in 5 mM K+ growth medium, a 4 fold greater blocking effect on sodium pumps after 2 days than expected from the association and dissociation rate constants measured in untreated or previously ouabain-treated cells. 4. This effect was: (a) approximately the same over a range of external potassium concentrations from 0.3 to 5 mM, although the absolute effect of ouabain over this range of potassium was much different; (b) probably not due to different isoforms of pumps in cells grown in ouabain compared to untreated cells; (c) apparently not a consequence of internalisation of pump-glycoside complexes. 5. We conclude that ouabain has only a limited access to sodium pumps in whole cells; this could be because sodium pumps cycle continuously through an inaccessible region of the plasma membrane. This effect needs to be considered both in the assessment of the magnitude of the long term effects of cardiac glycosides on cells, and in the measurement of the glycoside affinities of various isoforms of the pump.

Cardiac Glycosides↗

PubMed1990

Localisation and characterisation of functional vasoactive intestinal peptide receptors in feline kidney.

Specific 125I-labelled vasoactive intestinal peptide (VIP) binding was determined in feline renal cortical and medullary plasma membranes. For the cortex, Scatchard analysis of the data resulted in a curvilinear plot with a high-affinity site K0.5 of 8.4 +/- 2.6 nmol l-1 (SE, n = 6) and a second low-affinity site K0.5 204 +/- 16 nmol l-1 with binding site concentrations (Bmax) of 385 +/- 44.5 and 2710 +/- 181.3 fmol mg protein-1 respectively. Conversely a similar analysis of the results obtained for outer medullary membranes gave a single site with a K0.5 of 1.2 +/- 0.2 nmol l-1 (SE, n = 4) and Bmax of 157.8 +/- 24.7 fmol mg-1. Inner medullary membrane binding data. Gave a single site of lower affinity (K0.5 = 62.5 +/- 21.6 nmol l-1; n = 3). Structurally related peptides, glucagon and secretin, were ineffective (up to 1 mumol l-1) in displacing VIP from specific sites in both cortex and medulla. Porcine PHI 1-27 (a peptide having N-terminal histidine and C-terminal isoleucine) and a VIP antagonist [4-Cl-D-Phe6Leu17]VIP both displaced 125I-VIP from cortical and medullary membrane binding sites with IC50 values of 43.0 nmol l-1 and 1.3 mumol l-1 (cortex) and 132.0 nmol l-1 and 1.5 mumol l-1 (medulla) respectively. The localisation of specific VIP binding sites in feline kidney was investigated further by in vitro autoradiography.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenylyl Cyclases↗

PubMed1990

Vasoactive intestinal peptide stimulation of adenylate cyclase activity in isolated rabbit renal arterial microvessels.

Vasoactive intestinal peptide (VIP) at a concentration of 1 microM stimulates adenylate cyclase (AC) in isolated rabbit renal preglomerular arterial vessels by 3.9 +/- 1.3 fold (n = 6) over basal values. A comparable stimulation of AC was observed with prostaglandin E1. Half-maximal VIP-stimulated activity was observed at 7.2 +/- 3.5 nM (n = 6), which was increased to 17.4 +/- 4.4 nM by 10 microM-(4Cl-D-Phe6,Leu17)VIP, a VIP-receptor antagonist.

Adenylyl Cyclases↗