Biomedical subjects

J M Gee

Publications and source records attributed to J M Gee.

At least 91 records · Page 5Linked to original sources

PubMed1990

An immunocytochemical method for localization of estrogen receptors in rat tissues using a dinitrophenyl (DNP)-labeled rat monoclonal primary antibody.

We have developed an immunocytochemical method to demonstrate estrogen receptor in hormone-sensitive tissues of the rat using a dinitrophenyl (DNP) hapten-labeled rat antihuman estrogen receptor monoclonal antibody (MAb), H222. Mouse IgM anti-DNP was used secondarily, followed by a DNP/peroxidase conjugate, diaminobenzidine/hydrogen peroxide chromogen, and silver intensification. This method was applied to tissues from intact female rats and showed that estrogen receptor was localized in the nuclei of the stromal and glandular components of the uterine endometrium. Reduced receptor staining was observed in the luminal epithelium, with minimal myometrial staining. Anterior pituitary glands showed heterogeneous immunostaining and ovaries expressed the receptor predominantly in the interstitial cells; fallopian tubes demonstrated substantial epithelial staining. Uteri from chemically castrated rats showed reduced estrogen receptor immunostaining in both stromal and luminal cells, whereas staining was enhanced in the glandular elements. Classical estrogen-unresponsive tissues (heart, lung, and spleen) were unstained. Antibody controls involved pre-blocking antibody recognition sites on the receptor with unlabeled antibodies to estrogen receptor (H222, H226, and D547), as well as use of an inappropriate DNP-labeled antibody to metallothionein. These controls illustrated the specific nature of the DNP-H222 binding.

Adnexa Uteri↗

PubMed1989

Effect of oat gum on the physical properties of the gastrointestinal contents and on the uptake of D-galactose and cholesterol by rat small intestine in vitro.

Recent reports indicate that oats have a relatively low glycaemic effect in comparison with other carbohydrate food, and that their consumption leads to a reduction in plasma-cholesterol levels in man. These properties may be due to a soluble non-starch polysaccharide in oats. The present study was undertaken to explore the physiological properties of this material. Three groups of male Wistar rats were meal-fed on a control diet free of soluble dietary fibre for 10 d before being given a 10 g meal of either the control diet, a diet containing oat gum (beta-glucan), or finely ground rolled oats. The contents of the stomach, small intestine and caecum were later recovered and the weight, water content and viscosity were measured. The small intestinal contents from oat-gum-fed or oat-fed rats had a higher wet: dry weight ratio than that of the controls, and a higher viscosity. In in vitro studies the rate of uptake of D-galactose by jejunal rings was reduced in the presence of oat gum. The estimated Michaelis-Menten constant for the carrier-mediated component in the presence of oat gum was higher than that for controls, but the maximum transport rates were similar. Cholesterol uptake by everted jejunal sacs was progressively inhibited by increasing concentrations of oat gum in the mucosal medium. It is concluded that increased viscosity of the contents of the small intestine may contribute to the low glycaemic index and hypocholesterolaemic effects of oats in man. Oats appear to be amongst the few palatable sources of viscous dietary fibre in the conventional Western diet.

Animals↗

PubMed1988

The effect of Gypsophila saponins in the diet on mineral status and plasma cholesterol concentration in the rat.

1. Immature, male Wistar rats were allocated to one of six groups and caged individually. The first group was given a semi-synthetic diet containing 38 mg iron and 55 mg zinc/kg (basal group). The second and third groups were given a diet containing 10 mg Zn and 12 mg Fe/kg respectively (low-Zn and low-Fe groups). Groups four, five and six were given similar diets containing 20 g Gypsophila saponins/kg. After 21 d the Fe and Zn status of the rats was estimated and plasma cholesterol concentration determined. 2. Measurements of whole blood haemoglobin concentration, packed cell volume and liver Fe stores indicated that rats in the basal + saponin and low-Fe + saponin groups had a significantly reduced Fe status when compared with their controls. Rats in the low-Zn + saponin group also showed a trend toward reduced Fe stores. 3. Zn status, as judged by femur Zn concentration, was not adversely affected by the inclusion of Gypsophila saponins in the diet. 4. Consumption of the saponins resulted in a significant reduction in blood cholesterol concentration, with rats in both the low-Fe groups having significantly lower concentrations than their basal and low-Zn counterparts. 5. In view of suggestions that the consumption of saponins should be encouraged because of their ability to lower blood cholesterol, possible effects on Fe metabolism should be investigated further, particularly with respect to the levels and sources of saponin in the human diet.

Animals↗

PubMed1988

Intestinal microflora and gastrointestinal adaptation in the rat in response to non-digestible dietary polysaccharides.

1. A comparison was made of the effect of a fibre-free diet and diets containing non-digestible polysaccharides on rat caecal and colonic physiology and microflora. 2. All polysaccharide-containing diets led to enlargement of the caecum and colon, associated with increased weight of contents, and of tissue. Carboxymethylcellulose (CMC) had the most marked effect and animals given this also had watery faeces. 3. The density of bacteria in the caecum and colon varied significantly with diet and the proportion of aerobic bacteria in the flora was increased by the CMC diet. 4. In vitro, CMC and hydroxypropylmethylcellulose were poorly fermented. 5. There was a high correlation (caecum r 0.93; colon r 0.94) between tissue weight and wet weight of organ contents but no correlation with bacterial density, number of bacteria per organ, moisture content or short-chain fatty acid content. 6. It is concluded that caecal and colonic enlargement is due to tissue hypertrophy in response to increased bulk of contents, irrespective of the nature of that bulk which varies with diet; it is unlikely that short-chain fatty acids or other microbial metabolites are the stimulus for the trophic response seen when non-digestible dietary polysaccharides are fed to rats.

Adaptation, Physiological↗

PubMed1988

Plasma enteroglucagon and small bowel cytokinetics in rats fed soluble nonstarch polysaccharides.

The influence of dietary fiber on plasma glucagon and enteroglucagon (EG) and on ileal crypt cell production rate (CCPR) was assessed in the rat. A fiber-free semisynthetic diet and diets containing insoluble cellulose, Na-carboxymethylcellulose (CMC), guar gum, pectin, gum arabic, methylcellulose (MC), and hydroxypropylmethylcellulose (HPMC) at 10 g/100 g were fed for 14 d. Animals given soluble polysaccharides had plasma EG levels significantly higher than animals given insoluble cellulose, but only those fed the most viscous gums showed evidence of increased mucosal cell proliferation in the distal ileum. It was concluded that viscous nonfermentable polysaccharide gums stimulate the release of EG in the rat by slowing the absorption of nutrient, whereas the nonviscous polysaccharide gum arabic, or its breakdown products, probably directly stimulate mucosal endocrine cells. The failure of gum arabic to stimulate CCPR despite increased plasma EG indicated that EG alone is not a sufficient stimulus for increased mucosal cell proliferation.

Animals↗

PubMed1988

Interactions between hemolytic saponins, bile salts and small intestinal mucosa in the rat.

The interaction between bile salts and saponins from Gypsophylla was investigated in vitro, using changes in transmural potential difference across isolated lengths of rat jejunum as an index of the capacity of the saponin to affect the permeability of intestinal mucosal cells. The addition of saponin (ca. 5 mM) to taurocholic acid in buffered saline (pH 7.4) led to a significant increase in viscosity, indicating the formation of a polymer. The viscosity of the solution was highest at low molar ratios of bile salt to saponin (0.5-2.0). Under these conditions there was an inhibition of the permeating effect of saponins on the gut, as judged by the rate of decline in transmural PD. There was no evidence of inflammation or functional damage to the jejunal mucosa of rats fed a diet containing Gypsophylla saponin (ca. 1.5% w/w) for 7 d but changes in villus morphology were observed. There was also evidence of an increased rate of mucosal cell proliferation. Serum cholesterol levels were significantly lower in saponin-fed rats than in controls, while the cholesterol content of the cecal contents was increased. These results suggest that despite the protective effect of bile salts, Gypsophylla saponins interact with the mucosa of the proximal small intestine in vivo, but at the dietary level used in this study the mucosa was protected by an enhanced rate of cell replacement. The loss of cholesterol via exfoliated mucosal cells may contribute to the hypocholesterolemic effect of saponins in rats.

Animals↗

PubMed1987

The effect of dietary protein source and guar gum on gastrointestinal growth and enteroglucagon secretion in the rat.

1. Male Wistar rats (approximately 100 g) were given fibre-free semi-synthetic diets containing either casein or albumin (168 g/kg diet) as the protein source with or without guar gum (75 g/kg diet) (casein, albumin, casein guar gum and albumin-guar gum groups). 2. Small intestinal length, weights of caecal tissue and contents and plasma enteroglucagon concentration were significantly increased in guar-gum-fed animals compared with the fibre-free groups. 3. Total caecal weight and plasma enteroglucagon concentration were higher in the albumin-guar gum group compared with the casein-guar gum group. The weights of caecal tissue and contents were significantly increased in rats given the fibre-free albumin diet compared with those consuming a similar diet with casein as the protein source, although daily food intake tended to be lower. 4. It is concluded that the effect of materials classed as dietary fibre may be significantly influenced by the non-polysaccharide component of the diet, and that such interactions may influence both the growth and endocrine activity of the gastrointestinal tract.

Albumins↗

PubMed1986

Hexose absorption from jejunal loops in situ in zinc-deficient and Zn-supplemented rats.

1. Immature, male Wistar rats were given a low-zinc semi-synthetic diet (2 mg Zn/kg) for 22-28 d. Control groups received a similar diet supplemented with 58 mg Zn/kg either ad lib., or in amounts matched to the consumption of the Zn-deficient group. There was a rapid onset of reduced food consumption and growth retardation in the Zn-depleted animals. 2. Serosal surface area of small intestines taken from Zn-deficient rats was significantly reduced compared with that of control animals. Villi, dissected from samples of proximal jejunum, were markedly smaller than those of control rats and were present in greater numbers per unit area of serosa. 3. Luminal loss of galactose from jejunal loops in situ was significantly greater in the Zn-deficient rats compared with controls when expressed in terms of unit dry weight of intestine and serosal or villous surface area. Since only a small proportion of the total galactose remained in the mucosal tissue and associated extracellular space, this loss could only be accounted for by an increased efficiency of net trans-epithelial transport. Differences in total galactose absorption per unit length of jejunum were not so marked. 4. This intestinal adaptation to Zn-deficiency allows the maintenance of normal, and possibly increased, rates of hexose transfer into the body of animals exhibiting severe growth retardation, reduced food utilization and abnormal glucose metabolism.

Animals↗

PubMed1986

Gastrointestinal adaptation in response to soluble non-available polysaccharides in the rat.

1. Rats were fed on a control semi-synthetic diet containing insoluble cellulose (Solkafloc; 100 g/kg; control group) as the only source of dietary fibre, or on one of two test diets containing the same quantity of either guar gum or carboxymethylcellulose (CMC). Animals in the test groups showed similar growth rates and food intakes, which were significantly lower than those of the control group. The CMC group produced frequent poorly formed faeces throughout the 21 d feeding period. 2. The small intestines of animals in both test groups were significantly longer than those of the control group at the end of the study. The caeca were also enlarged and heavier, particularly in the CMC-fed group. 3. The rate of production of mucosal cells was increased in the small and large intestines of both test groups. The CMC-fed group exhibited a particularly high rate in the distal ileum, where the rate of cell divisions per crypt was over three times greater than at the same site in the control group. The increased proliferation was associated with a significant lengthening of the crypts and an approximately 25% increase in the basal width of the villi. 4. Mucosal alkaline phosphatase (EC 3.1.3.1) and lactase (EC 3.2.1.23) levels were lower than those of the control group at proximal and distal sites in the small intestines of both CMC- and guar-gum-fed groups. Altered spatial distributions of maltase (EC 3.2.1.20) and sucrase (EC 3.2.1.48) activities were also observed in these animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Physiological↗

PubMed1986

Intestinal microflora, morphology and enzyme activity in zinc-deficient and Zn-supplemented rats.

1. Immature, male Wistar rats were given a low-zinc diet (2 mg/kg) for 22-24 d. Control groups received a similar diet supplemented with 58 mg Zn/kg either ad lib., or in amounts matched to the consumption of the Zn-deficient group. Food consumption, rate of growth and food conversion efficiency were markedly lower in the Zn-deficient group of rats compared with controls. Appetite, growth rate and food utilization improved dramatically over a subsequent 4 d period of Zn supplementation. 2. Morphological examination of samples of jejunum and ileum confirmed that Zn deficiency in the rat is accompanied by a reduction in villous dimensions and increase in villous density. After a short period of Zn supplementation, villous density and the basal width and maximum height of individual villi in the jejunum returned to normal. Similar changes occurred in the ileum but to a lesser extent. 3. Mucosal alkaline phosphatase (EC 3.1.3.1) activity was significantly lower in the small intestine of Zn-deficient rats compared with Zn-supplemented rats. Disaccharidase activities were lower in the Zn-deficient group, compared with their feed-restricted counterparts, but were similar to values for ad lib.-fed controls. Tissue alkaline phosphatase and disaccharidase activities were consistently higher after a 4 d period of Zn supplementation, compared with non-supplemented animals, but this increase was only significant for alkaline phosphatase. 4. Although there were striking similarities in the mucosal characteristics of gnotobiotic and Zn-deficient rats, there was no indication that even severe dietary Zn depletion reduced the numbers of viable bacteria present in either the small or large intestine.(ABSTRACT TRUNCATED AT 250 WORDS)

Alkaline Phosphatase↗

PubMed1986

Influence of saponins on gut permeability and active nutrient transport in vitro.

The influence of four saponins, three triterpenoid glycosides and one steroidal amine glycoside, upon intestinal transport was investigated in vitro. In the presence of Gypsophylla saponin, carrier-mediated galactose transport was inhibited, although the uptake of the passively transported L-isomer of glucose increased. The uptake of the extracellular space marker, polyethylene glycol 4000, was also higher, indicating that the saponin inhibited active transport by increasing the general permeability of the enterocytes. Gypsophylla saponin, in contact only with the mucosal surface of everted jejunal sacs, induced a rapid decline in glucose-stimulated transmural potential difference. The rate of decline increased as the saponin concentration was raised over the approximate range of 0.3 to 8 mM. Saponaria saponin and alpha-tomatine also reduced transmural potential difference, but soya saponins were much less effective. The results indicate that some saponins readily increase the permeability of the small intestinal mucosal cells, thereby inhibiting active nutrient transport, and facilitating the uptake of materials to which the gut would normally be impermeable.

Animals↗

PubMed1985

Differences in intestinal protein synthesis and cellular proliferation in well-nourished rats consuming conventional laboratory diets.

Male Wistar rats (100 g) were given a commercial pellet feed or a semi-synthetic diet ad lib. Although the pellet-fed group grew slightly faster than the other group during the early part of the feeding period, there was no significantly difference between the final weights of the groups. The fractional rates of protein synthesis in jejunum, proximal ileum and liver were measured by a technique based on the determination of L-[4-3H]phenylalanine incorporation over a short time period. Protein synthesis was higher in both jejunum and ileum of the pellet-fed rats compared with those eating the semi-synthetic diet, but there was no difference between the rates of protein synthesis measured in the livers of the groups. The rate of mucosal cell division was significantly faster in the ileal mucosa of the pellet-fed group compared with the other group, and there were significant differences in some aspects of mucosal morphology. The maintenance of higher rates of cell turnover and protein synthesis in animals given a commercial pellet feed is unexplained, but it may be related to the presence of non-absorbable polysaccharides or other complex plant materials in the pellet feed.

Animal Feed↗

PubMed1985

Intestinal cellular proliferation and protein synthesis in zinc-deficient rats.

Immature male Wistar rats were given a low-zinc semi-synthetic diet (2 mg Zn/kg) for 28 d. Control groups received a similar diet supplemented with 58 mg Zn/kg either ad lib. or in amounts matched to the consumption of the Zn-deficient group. Rates of growth, food consumption and small intestinal length were significantly reduced in the Zn-depleted rats. Zn deficiency in the rat was associated with a reduction in the ratio, crypt: villus and a lower rate of crypt cell division in the jejunum. This resulted in a substantial decrease in the net influx of new cells into the villi of the Zn-deficient animals compared with controls. The fractional rates of protein synthesis in jejunal mucosa were measured by a technique based on the determination of L-[4-3H]phenylalanine incorporation. There was no evidence of a decline in the protein synthetic rate in total mucosa from Zn-deficient rats. It is suggested that a reduction in cell influx into the villi may be responsible for the morphological and functional changes observed in the small intestine of rats fed on a low-Zn diet.

Animals↗

PubMed1984

Hexose transport and mucosal morphology in the small intestine of the zinc-deficient rat.

Immature male Wistar rats were given a zinc-depleted semi-synthetic diet (2 mg Zn/kg diet) for 4 weeks. Control groups received a similar diet containing 60 mg Zn/kg diet, either ad lib. or in amounts matched to the consumption of the Zn-deficient group. Rates of growth and food conversion efficiency were markedly lower in the Zn-deficient group compared to controls. The phloridzin-sensitive uptake of galactose and 3-O-methyl glucose by everted jejunal rings from each group was measured over a range of concentrations. Kinetic analysis revealed that the maximum transport rate (Vmax) for the uptake of both sugars was significantly higher in the Zn-deficient group than in the control groups. The villi, microdissected from samples of proximal jejunum, were significantly shorter and narrower across the base in the Zn-deficient group than those of the control groups, but were present in greater numbers per unit serosal area. The present study shows that Zn deficiency leads to significant morphological changes in rat small intestine, which are accompanied by an increase in the capacity of the tissue to take up sugars across the mucosal membrane.

3-O-Methylglucose↗

PubMed1984

Effect of dietary supplements of guar gum and cellulose on intestinal cell proliferation, enzyme levels and sugar transport in the rat.

Male Wistar rats (approximately 200 g) were given fibre-free semi-synthetic diets containing either sucrose (S) or a sucrose-starch mixture (SS) as the carbohydrate component, or a diet similar to SS containing 40 g guar gum/kg (G), or 100 g cellulose/kg (C). The animals remained healthy, and weight gain after 30 d was similar in all groups. The small intestines of the animals given diet G were significantly longer than those of the other groups, and showed signs of increased mitotic activity and mucosal growth. No significant differences in mucosal enzyme activity were detected between the two fibre-free control groups. Lactase (EC 3.2.1.23) and alkaline phosphatase (EC 3.1.3.1) activities were significantly lower than controls in group G, but were higher in group C. Kinetic analysis of 3-O-methyl glucose uptake by isolated intestine indicated that the maximum transport rate (Vmax) of tissue from group G tended to be lower than from the fibre-free group SS and group C. It is concluded that materials which are classed as dietary fibre but which differ markedly in their physical properties may also differ in the functional changes to which they give rise in the small intestine. These changes may be at least partially mediated by effects on mucosal cell proliferation.

3-O-Methylglucose↗

PubMed1983

The influence of cooked kidney beans (Phaseolus vulgaris) on intestinal cell turnover and faecal nitrogen excretion in the rat.

Male Wistar rats were fed on semi-synthetic diets containing cooked white kidney beans (Phaseolus vulgaris) or equivalent levels of protein and carbohydrate. No change was observed in over-all nitrogen balance in animals fed on the bean diet, but there was a two- to three-fold increase in their faecal excretion, compared with control rats. This was compensated by a decrease in urinary-N excretion. Homogenized small intestinal mucosa, prepared from bean-fed animals, showed a 28% increase in protein content compared with control material. Measurements of 3H-labelled thymidine turnover indicated that mucosal cell exfoliation was increased by approximately 35% in the small intestines of bean-fed rats compared with controls. It is concluded that though a diet rich in cooked P. vulgaris leads to some increase in mucosal cell turnover in the small intestine of rats, the consequent increase in mucosal protein loss could not account for the increased faecal-N excretion seen in these animals.

Animals↗

PubMed1983

The influence of guar gum on intestinal cholesterol transport in the rat.

Everted sacs of rat proximal small intestine were used to determine the effect of guar gum (5 g/l) on the uptake of cholesterol (0.1 mM) from a solution of micelles. The uptake of cholesterol was found to be linear both in the presence and absence of guar gum. When guar was present throughout the whole of the incubation medium, the uptake of cholesterol was reduced to approximately 40% of control values. Sacs which had been pre-incubated in guar gum before exposure to cholesterol in a guar-free medium also showed a reduction in cholesterol uptake but this was less pronounced. A two-stage perfusion technique, previously described (Blackburn & Johnson, 1981), was used to determine the effect of a guar layer adsorbed to the mucosal surface on cholesterol absorption in vivo. Such a layer leads to a reduction of approximately 36%; it was concluded that guar slows the absorption of cholesterol from micelles by a mechanism, or mechanisms, involving an increased resistance to diffusion in the aqueous medium. Groups of rats were meal-fed for at least 30 d on semi-synthetic diets with or without the inclusion of guar gum (20 g/kg). Rates of intestinal absorption of cholesterol, glucose and fluid were then determined by the perfusion technique in vivo. There was no reduction in absorption in the test animals compared with the controls. It is proposed that guar gum is able to slow the intestinal transport of cholesterol from a suspension of pre-formed micelles, but only when both are present in the lumen together. No evidence was obtained to suggest that the consumption by rats of a diet containing guar gum, at a level similar to that used in human studies, leads to any adaptive reduction in their rates of cholesterol or glucose absorption.

Animals↗

PubMed1982

Influence of viscous incubation media on the resistance to diffusion of the intestinal unstirred water layer in vitro.

Everted sacs of rat intestine were incubated in media made viscous with Na-carboxymethylcellulose (CMC). This increased the resistance to diffusion of the mucosal unstirred water layer, as measured by the half-time for the establishment of osmotic streaming potentials. The increase ws not however linearly related to viscosity, but reached a maximum, equivalent to an apparent thickness of 500 mu, at a viscosity of about 100 cP (shear rate = 50 s-1). The inhibition of maltose hydrolysis by CMC conformed to theoretical expectations, and was characterised by an increased apparent Km, with no change in Vmax. This was not true however of galactose transport by everted sacs which showed a decline in maximum transport rate, with no apparent increase in Km. It appears that increased unstirred layer resistance may induce partial anoxia in the everted sac preparation and thus reduce the energy available for active transport. We conclude that the effect of enhanced viscosity is to reduce turbulent flow, and hence stirring, in the intervillous compartment. Such an effect may be brought about by viscous components of the diet and may influence rates of absorption and digestion in the intact intestine in vivo.

Animals↗