Absorption of beta-casomorphins from autoperfused lamb and piglet small intestine

1990 ◽  
Vol 259 (3) ◽  
pp. G443-G452 ◽  
Author(s):  
L. C. Read ◽  
A. P. Lord ◽  
V. Brantl ◽  
G. Koch
Keyword(s):  
Small Intestine ◽  
Intestinal Lumen ◽  
Physiological Conditions ◽  
Naturally Occurring ◽  
Gut Epithelium ◽  
Measured Absorption ◽  
Beta Casein ◽  
Kinetics Of

beta-Casomorphins (beta-CMs) derived from milk beta-casein may exert various opiate activities in milk-fed infants. To assess the physiological significance of beta-CMs as a source of circulating opioids in infants, we measured absorption rates of several beta-CMs under near-physiological conditions using in situ autoperfused lamb intestine. The naturally occurring beta-CMs, beta-CM-7 and beta-CM-4-amide, were absorbed readily into blood with no transfer into lymph. Uptake peaked within several minutes of the luminal infusion of peptide but then declined sharply and stopped within a further 10-15 min. The recovery in blood, intestinal contents, and tissue at the end of the 30-min experiment was less than 1% of the infused dose. The low recovery was due to rapid proteolysis based on in vitro studies that demonstrated half-lives of less than 5 min in lamb blood, luminal contents, and lymph. The synthetic dipeptidyl peptidase IV-resistant analogue beta-[D-Ala2]CM- 4-amide was stable during incubation in blood, lymph, or luminal contents and was absorbed into blood at rates that were maximal within several minutes and remained steady for the 30-min period. We conclude that although natural beta-CMs are transferred across the lamb small intestine, rapid degradation within the intestinal lumen, gut epithelium, and blood would prevent entry into the circulation under normal conditions. Val-beta-CM-7, a putative stable precursor, had similar stability and kinetics of absorption to beta-CM-7, results that exclude Val-beta-CM-7 as a stable precursor for delivery of beta-CMs to the circulation. Essentially identical results to those in lambs were obtained in 7-day-old piglets.

scholarly journals Part of quercetin absorbed in the small intestine is conjugated and further secreted in the intestinal lumen

1999 ◽  
Vol 277 (1) ◽  
pp. G120-G126 ◽  
Author(s):  
Vanessa Crespy ◽  
Christine Morand ◽  
Claudine Manach ◽  
Catherine Besson ◽  
Christian Demigne ◽  
...  
Keyword(s):  
Small Intestine ◽  
Intestinal Wall ◽  
Intestinal Lumen ◽  
Intestinal Cells ◽  
Biliary Duct ◽  
In Situ Perfusion ◽  
Perfusion Period ◽  
Hydrolysis Of

Rutin and quercetin absorption and metabolism were investigated in rats after in situ perfusion of jejunum plus ileum (15 nmol/min). In contrast to rutin, a high proportion of quercetin (two-thirds) disappeared during perfusion, reflecting extensive transfer into the intestinal wall. Net quercetin absorption was not complete (2.1 nmol/min), inasmuch as 52% were reexcreted in the lumen as conjugated derivatives (7.7 nmol/min). Enterohepatic recycling contribution of flavonoids was excluded by catheterization of the biliary duct before perfusion. After a 30-min perfusion period, 0.71 μM of quercetin equivalents were detected in plasma, reflecting a significant absorption from the small intestine. The differential hydrolysis of effluent samples by glucuronidase and/or sulfatase indicates that the conjugated forms released in the lumen were 1) glucuronidated derivatives of quercetin and of its methoxylated forms (64%) and 2) sulfated form of quercetin (36%). In vitro quercetin glucuronides synthetized using jejunal and ileal microsomal fractions were similar to those recovered in the effluent of perfusion. These data suggest that glucuronidation and sulfatation take place in intestinal cells, whereas no glucurono-sulfoconjugates could be detected in the effluent. The present work shows that a rapid quercetin absorption in the small intestine is very effective together with its active conjugation in intestinal cells.

Changes in lipophosphoglycan and gene expression associated with the development ofLeishmania majorinPhlebotomus papatasi

Parasitology ◽  
1995 ◽  
Vol 111 (3) ◽  
pp. 275-287 ◽  
Author(s):  
E. M. B. Saraiva ◽  
P. F. P. Pimenta ◽  
T. N. Brodin ◽  
E. Rowton ◽  
G. B. Modi ◽  
...  
Keyword(s):  
Differential Expression ◽  
Fold Increase ◽  
Surface Expression ◽  
Side Chain ◽  
Surface Coat ◽  
Natural Vector ◽  
Metacyclic Promastigotes ◽  
Kinetics Of

SUMMARYStage-specific molecular and morphogenic markers were used to follow the kinetics of appearance, number, and position of metacyclic promastigotes developing during the course ofL. majorinfection in a natural vector,Phlebotomus papatasi. Expression of surface lipophosphoglycan (LPG) on transformed promastigotes was delayed until the appearance of nectomonad forms on day 3, and continued to be abundantly expressed by all promastigotes thereafter. An epitope associate with arabinose substitution of LPG side-chain oligosaccharides, identified by its differential expression by metacyclics invitro, was detected on the surface of a low proportion of midgut promastigotes beginning on day 5, and on up to 60% of promatigotes on days 10 and 15. In contrast 100% of the parasites egested from the mouthparts during forced feeding of 15 day infected flies stained strongly for this epitope. At each time-point, the surface expression of the modified LPG was restricted to morphologically distinguished metacyclic forms. Ultrastructural study of the metacyclic surface revealed an approximate 2-fold increase in the thickness of the surface coat compared to nectomonad forms, suggesting elongation of LPG as occurs during metacyclogenesisin vitro. A metacyclic-associated transcript (MAT-1), another marker identified by its differential expression invitro, also showed selective expression by promastigotes in the fly, and was used inin situhybridization studies to demonstrate the positioning of metacyclics in the anterior gut.

Demonstration and modification of intervillous pH profiles in rat small intestine in vitro

1989 ◽  
Vol 257 (4) ◽  
pp. G489-G495 ◽  
Author(s):  
H. Daniel ◽  
C. Fett ◽  
A. Kratz
Keyword(s):  
Small Intestine ◽  
Low Ph ◽  
Rat Small Intestine ◽  
Ph Profile ◽  
Subsequent Increase ◽  
Alkaline Secretion ◽  
Alkaline Fluid ◽  
Ph Profiles

The intervillous pH profiles along the crypt villus axis in different regions of the rat small intestine were measured in vitro by using pH-sensitive liquid ion-exchanger microelectrodes. A characteristic pH profile was observed in the duodenum and jejunum. A region of low pH was detected in the upper parts of the villi (pH 6.65 +/- 0.06 to 6.85 +/- 0.07), whereas pH at the villus base was always higher. In the ileum no gradient was observed (pH 7.26 +/- 0.05 to 7.31 +/- 0.05). Preincubation of the tissue in situ with 10 mM theophylline for 1 h caused an increase in the villus base pH in the jejunum (pH 7.24 +/- 0.04) and ileum (7.44 +/- 0.04) followed by a subsequent increase of the pH in the upper part of the villi. These results indicate that the low pH in the upper intervillous space may be related to H+ secretion occurring from the mature enterocytes, whereas the crypt cells may secrete a rather neutral or slightly alkaline fluid. Alkaline secretion from the crypts may be increased by theophylline, which changes the levels of cyclic nucleotides in the mucosa.

scholarly journals Comparative Survival and the Cold-Induced Gene Expression of Pathogenic and Nonpathogenic Vibrio Parahaemolyticus from Tropical Eastern Oysters during Cold Storage

2020 ◽  
Vol 17 (6) ◽  
pp. 1836
Author(s):  
Francisco Alarcón Elvira ◽  
Violeta T. Pardío Sedas ◽  
David Martínez Herrera ◽  
Rodolfo Quintana Castro ◽  
Rosa María Oliart Ros ◽  
...  
Keyword(s):  
Low Temperatures ◽  
Vibrio Parahaemolyticus ◽  
Cold Shock ◽  
Gene Expressions ◽  
Reverse Transcription Pcr ◽  
Naturally Occurring ◽  
Gene Response ◽  
Eastern Oysters ◽  
Kinetics Of

Expression of the regulatory stress rpoS gene controls the transcription of cspA genes, which are involved in survival and adaptation to low temperatures. The purpose of this study was to assess the growth kinetics of naturally occurring V. parahaemolyticus in shellstock oysters and in vitro and the cold-shock-induced expression of the rpoS and cspA gene response in vitro during postharvest refrigeration. Naturally contaminated eastern oysters (Crassostrea virginica) and pathogenic (Vp-tdh) and nonpathogenic (Vp-tlh) isolates were stored at 7 ± 1 °C for 168 h and 216 h, respectively. The regulatory stress (rpos) and cold-shock (cspA) gene expressions were determined by reverse transcription PCR. At 24 h, the (Vp-tdh) strain grew faster (p < 0.05) than the (Vp-tlh) strain in oysters (λ = 0.33, 0.39, respectively) and in vitro (λ = 0.89, 37.65, respectively), indicating a better adaptation to cold shock for the (Vp-tdh) strain in live oysters and in vitro. At 24 h, the (Vp-tdh) strain rpoS and cspA gene expressions were upregulated by 1.9 and 2.3-fold, respectively, but the (Vp-tlh) strain rpoS and cspA gene expressions were repressed and upregulated by −0.024 and 1.9-fold, respectively. The V. parahaemolyticus strains that were isolated from tropical oysters have adaptive expression changes to survive and grow at 7 °C, according to their virulence.

scholarly journals Synthetic and genetic dimers as quantification ruler for single-molecule counting with PALM

2019 ◽  
Vol 30 (12) ◽  
pp. 1369-1376 ◽  
Author(s):  
Tim N. Baldering ◽  
Marina S. Dietz ◽  
Karl Gatterdam ◽  
Christos Karathanasis ◽  
Ralph Wieneke ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Single Molecule ◽  
Fusion Proteins ◽  
Fluorescent Proteins ◽  
Superresolution Microscopy ◽  
Superresolution Imaging ◽  
Molecular Information ◽  
Kinetics Of

How membrane proteins oligomerize determines their function. Superresolution microscopy can report on protein clustering and extract quantitative molecular information. Here, we evaluate the blinking kinetics of four photoactivatable fluorescent proteins for quantitative single-molecule microscopy. We identified mEos3.2 and mMaple3 to be suitable for molecular quantification through blinking histogram analysis. We designed synthetic and genetic dimers of mEos3.2 as well as fusion proteins of monomeric and dimeric membrane proteins as reference structures, and we demonstrate their versatile use for quantitative superresolution imaging in vitro and in situ. We further found that the blinking behavior of mEos3.2 and mMaple3 is modified by a reducing agent, offering the possibility to adjust blinking parameters according to experimental needs.

Kinetics of transepithelial movement of heavy metals in rat jejunum

1987 ◽  
Vol 253 (2) ◽  
pp. G134-G138
Author(s):  
E. C. Foulkes ◽  
D. M. McMullen
Keyword(s):  
Heavy Metals ◽  
Venous Blood ◽  
Compartment Model ◽  
Intestinal Lumen ◽  
Rat Jejunum ◽  
Temperature Dependent ◽  
Appearance Time ◽  
Portal Venous Blood ◽  
Kinetics Of

The kinetics of the transepithelial movement of heavy metals were studied in the perfused rat jejunum in situ. The peak appearance time (TET) of Zn, Ni, and Cd in portal venous blood was determined after their transient (10 s) introduction into the intestinal lumen. Observed TET values for these metals were positively correlated with their affinities for metallothionein and agreed with those predicted on the basis of a linear three-compartment model that does not allow for paracellular pathways. Further evidence was provided to support the hypothesis that Cd absorption consists first of Cd binding to negative membrane charges followed by a temperature-dependent internalization step.

scholarly journals The Effect of Potassium on the Intestinal Transport of Glucose

1966 ◽  
Vol 50 (1) ◽  
pp. 113-128 ◽  
Author(s):  
T. Z. Csáky ◽  
P. M. Ho
Keyword(s):  
Small Intestine ◽  
Specific Activity ◽  
Lactic Acid Production ◽  
Sugar Transport ◽  
Intestinal Transport ◽  
Intestinal Lumen ◽  
High K ◽  
Rate Of Absorption ◽  
Specific Sugar

The rate of absorption of glucose, galactose, and 3-0-methylglucose was studied in the rat's small intestine perfused in situ with isosmotic solutions containing these sugars and Na2SO4 or K2SO4. The presence of high [K+] in the lumen enhances absorption of glucose but not that of galactose or of 3-0-methylglucose. The potassium stimulation is apparent at higher glucose concentrations where primarily carrier-mediated diffusion is involved in the translocation. In this case potassium stimulates transport even if it is the only cation in the lumen. The potassium-stimulated intestine produces more glycogen with higher specific activity than the control gut. Lactic acid production by the intestine is markedly enhanced if the intestinal lumen is perfused with a solution containing glucose and high [K+]. It is concluded that potassium does not affect permeability or the specific sugar transport system of the gut, but enhances intracellular metabolic disappearance of glucose thereby creating a larger luminal intracellular concentration gradient which in turn enhances the rate of carrier-facilitated entry.

scholarly journals DES2 protein is responsible for phytoceramide biosynthesis in the mouse small intestine

2004 ◽  
Vol 379 (3) ◽  
pp. 687-695 ◽  
Author(s):  
Fumio OMAE ◽  
Masao MIYAZAKI ◽  
Ayako ENOMOTO ◽  
Minoru SUZUKI ◽  
Yusuke SUZUKI ◽  
...  
Keyword(s):  
Small Intestine ◽  
Epithelial Cells ◽  
Intestinal Epithelial ◽  
Vitro Assay ◽  
Peptide Antibody ◽  
Mouse Small Intestine ◽  
Mrna Content ◽  
Crypt Cells

The C-4 hydroxylation of sphinganine and dihydroceramide is a rate-limiting reaction in the biosynthesis of phytosphingolipids. Mouse DES1 (MDES1) cDNA homologous to the Drosophila melanogaster degenerative spermatocyte gene-1 (des-1) cDNA leads to sphingosine Δ4-desaturase activity, and another mouse homologue, MDES2, has bifunctional activity, producing C-4 hydroxysphinganine and Δ4-sphingenine in yeast [Ternes, Franke, Zahringer, Sperling and Heinz (2002) J. Biol. Chem. 277, 25512–25518]. Here, we report the characterization of mouse DES2 (MDES2) using an in vitro assay with a homogenate of COS-7 cells transfected with MDES2 cDNA and N-octanoyl-sphinganine and sphinganine as substrates. MDES2 protein prefers dihydroceramide as a substrate to sphinganine, and exhibits dihydroceramide Δ4-desaturase and C-4 hydroxylase activities. MDES2 mRNA content was high in the small intestine and abundant in the kidney. In situ hybridization detected signals of MDES2 mRNA in the crypt cells. Immunohistochemistry using an anti-MDES2 peptide antibody stained the crypt cells and the adjacent epithelial cells. These results suggest that MDES2 is the dihydroceramide C-4 hydroxylase responsible for the biosynthesis of enriched phytosphingoglycolipids in the microvillous membranes of intestinal epithelial cells.

scholarly journals A comparative study of ruminal activity in Churra and Merino sheep offered alfalfa hay

1997 ◽  
Vol 65 (1) ◽  
pp. 121-128 ◽  
Author(s):  
M. J. Ranilla ◽  
M. D. Carro ◽  
C. Valdés ◽  
F. J. Giráldez ◽  
S. López
Keyword(s):  
Cell Wall ◽  
Rumen Fluid ◽  
Merino Sheep ◽  
Rumen Ph ◽  
Sheep Rumen ◽  
Fermentation Parameters ◽  
Micro Organisms ◽  
Kinetics Of

AbstractA study was carried out to compare the fermentation parameters and kinetics of digestion of a range of different foods in the rumen of two breeds of sheep (Churra and Merino). Ten mature sheep (five Churra and five Merino), each fitted with a rumen cannula, were used in this study. In situ rumen degradability of both dry matter (DM) and cell wall was greater in Churra than in Merino sheep, the breed differences being significant for most of the foods used in the study (P < 0·05). These differences were greater when the foods had a higher cell wall concentration and this could be related to differences in the ruminal environment. However, when the foods were incubated with rumen fluid their in vitro organic matter (OM) degradability was similar in both breeds. Rumen pH was higher (P < 0·05) and ammonia concentrations were lower (P < 0·05) in Churra than in Merino sheep. Rumen volatile fatty acid concentrations tended to be higher in Merino than in Churra sheep, though differences were only significant just before feeding (P < 0·05). The ratio acetate: propionate was higher in the Churra than Merino breed before and 12 h after feeding (P < 0·05). Protozoa numbers in rumen liquid were similar for both genotypes. The greater degradation of forages in the rumen of Churra sheep is discussed in relation to the possible higher activity of fibre-degrading micro-organisms and the greater buffering capacity of the rumen contents against fermentation acids, which could result in more favourable conditions for the microbial degradation of foods in the rumen.

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