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Lateral intercellular space volume as a determinant of CFTR-mediated anion secretion across small intestinal mucosa.

Studies of full-thickness, small intestinal preparations have shown that maximal anion secretion [indexed by short-circuit current (I(sc))] during intracellular cAMP (cAMP(i)) stimulation is transient and followed by a decline toward baseline. Declining I(sc) is preceded by decreases in transepithelial conductance (G(t)), which in the small intestine reflects the lateral intercellular space (LIS) volume of the paracellular pathway. We hypothesized that decreases in LIS volume limit the magnitude and duration of cAMP(i)-stimulated anion secretion. Experimental manipulations to increase the patency of the LIS (assessed by G(t) and electron microscopy) were investigated for an effect on the magnitude of cAMP(i)-stimulated anion secretion (assessed by the I(sc) and isotopic fluxes) across murine small intestine. In control studies, changes of G(t) after cAMP(i) stimulation were associated with a morphological "collapse" of the LIS, which did not occur in intestine of CFTR-null mice. Removal of the outer intestinal musculature, exposure to a serosal hypertonic solution, or increased serosal hydrostatic pressure minimized reductions in G(t) and increased the cAMP(i)-stimulated I(sc) response. Increased I(sc) primarily resulted from increased Cl(-) secretion that was largely bumetanide sensitive. However, bumetanide-insensitive I(sc) was also increased, and similar increases occurred in the Na(+)-K(+)-2Cl(-) cotransporter (NKCC1)-null intestine, indicating that activities of non-NKCC1 anion uptake proteins are also affected by LIS volume. Thus LIS patency is an important determinant of the magnitude and duration of CFTR-mediated anion secretion in murine small intestine. Decreases in LIS volume may limit the pool of available anions to basolateral transporters involved in transepithelial secretion.

Animals↗

p53 and PTEN/MMAC1 mutational analysis of the small-intestinal cancer.

A 66-year-old female, suffering from small-intestinal cancer underwent resection of the small intestine. Genome DNAs were extracted from the patient's blood and small-intestinal cancer and were subjected to a polymerase chain reaction-single strand conformation polymorphism and nucleotide sequence analysis for exons of the p53 and PTEN/MMAC1 genes to search for any mutations. The sequence analysis revealed a point mutation of the p53 codon 93 in the cancer DNA; however, no mutation of the PTEN/MMAC1 gene was observed in either the blood or cancer DNA. The p53 mutation, therefore, seems to be related to tumour progression of small-intestinal cancer; however, no relationship was found between the PTEN/MMAC1 gene and the small-intestinal cancer.

Aged↗

Bacteriologic analysis of mucosal biopsy specimens for detecting small-intestinal bacterial overgrowth.

BACKGROUND: Although culture of luminal secretions is regarded as the most accurate diagnostic test for small-intestinal bacterial overgrowth, obtaining an aspirate is often difficult owing to the sparseness of luminal secretions present at the time of aspiration. Obtaining a mucosal biopsy specimen for bacteriologic analysis would overcome this problem. METHODS: Culture of small-intestinal and gastric aspirates and unwashed small-intestinal mucosal specimens was performed in 51 adult subjects investigated for small-intestinal overgrowth. RESULTS: Highly significant (r = 0.85-0.90; p < 0.0005) correlations were found between viable bacterial counts in small-intestinal luminal secretions and biopsy specimens. Small-intestinal bacterial overgrowth was present in 60.8% of subjects. When specimens weighing 4.0-84.0 mg were suspended in diluent, total aerobic and/or anaerobic bacterial counts > or = 10(2) CFU/ml were found to have 90.3% sensitivity and 100% specificity for small-intestinal bacterial overgrowth. CONCLUSION: Culture of an unwashed small-intestinal mucosal biopsy specimen is a useful alternative to culture of a small-intestinal aspirate for detecting subjects with small-intestinal bacterial overgrowth, especially when luminal secretions are scanty at the time of aspiration.

Adult↗

Use of small intestinal submucosa for corporal body grafting in cases of severe penile curvature.

PURPOSE: Small intestinal submucosa is a unique biomaterial that has been found to promote tissue specific regeneration in the urinary tract. We present our experimental and clinical experience with small intestinal submucosa (SurgiSis, Cook Biotech, Spencer, Indiana) for pediatric corporal body reconstruction. MATERIAL AND METHODS: A total of 20 Fischer rats underwent implantation of a 7 x 3 mm. small intestinal submucosa graft following excision of an ellipse of tunica albuginea and 14 control animals underwent tunical excision with reimplantation of this autologous segment. The animals were euthanized, and the penis was sectioned and histologically studied at intervals of 1, 2, 4, 6, 16 and 24 weeks. In 15 pediatric patients small intestinal submucosa was used for corporal body grafting. The grafting procedure was performed along the ventral (hypospadias cases) or dorsal (epispadias cases) surface of the corporal bodies. The tunica albuginea was incised full thickness at the point of maximal curvature down to the cavernosal tissue and the defect was filled with a single layer of small intestinal submucosa. RESULTS: Measurements of the animal small intestinal submucosa grafts did not reveal significant graft contraction through 6 months. There was no graft expansion or ballooning after pharmacological induction of an artificial erection. Histologically, marked inflammation at 1 week precipitously decreased to a normal appearing tunica albuginea at 3 and 6 months. In all clinical cases small intestinal submucosa was found to be technically easy to handle. Mean followup is 14 months (range 5 to 26). All patients have a straight phallus as documented by observation of spontaneous erections or artificial erection at the time of stage 2 hypospadias repair. No complications occurred. CONCLUSIONS: Small intestinal submucosa demonstrates tissue specific regeneration properties in the rat and human tunica albuginea. It is an off-the-shelf material that is safe, technically easy to use and readily available.

Adolescent↗

Studies on the intestinal flora. II. Bacterial flora of the small intestine in patients with gastrointestinal disorders.

The type and distribution of bacteria in the jejunal juice of patients with a variety of gastrointestinal conditions that might affect the small intestinal flora were examined. Bacterial colonization of the jejunum defined, in this context, as the occurrence of a bile salttolerant flora consisting of both aerobic and anaerobic bacteria qualitatively resembling that of faeces, was observed only in patients with some form of blind loop. Prominent among the bacteria isolated from these colonized juices were non-sporing anaerobic bacteria, most usually Bacteroides, able to hydrolyse bile salts. No simple correlation between the patient's fat excretion and bacterial colonization of the jejunum could be demonstrated.

Adolescent↗

Luminal immunity in small-intestinal bacterial overgrowth and old age.

BACKGROUND: The independent influences of small-intestinal bacterial overgrowth and old age on mucosal immunoglobulin production and secretion have not been assessed. This is an important issue, since luminal IgA deficiency may exacerbate small-intestinal bacterial overgrowth, the prevalence of which is high in selected elderly populations. METHODS: Proximal small-intestinal aspirates were obtained from 33 subjects for bacteriologic analysis and measurement of total IgA, IgM, total IgG. IgG subclass, and IgD concentrations. IgA subclasses were measured in 24 unselected subjects. Serum immunoglobulin and salivary IgA concentrations were measured in all subjects. RESULTS: IgA2 and IgG3 were predominant IgA and IgG subclasses in proximal small-intestinal luminal secretions. Luminal concentrations of IgA2 and IgM, but not IgG3 or any other IgG subclass, were significantly increased in small-intestinal bacterial overgrowth, which was present in 19 of 33 (57.6%) subjects. Old age did not influence these levels. Luminal immunoglobulin concentrations did not correlate significantly with either serum or salivary values. IgD was not measureable in proximal small-intestinal secretions. CONCLUSIONS: Increased luminal concentrations of the secretory immunoglobulins, IgA2 and IgM, occur in small-intestinal bacterial overgrowth. Local investigation is mandatory when assessing the mucosal immunopathology of this disorder. Luminal IgG3 is unlikely to be predominantly derived from serum. Old age does not independently influence luminal immunity.

Age Factors↗

Intestinal bypass. A comparison between two different bypass operations and resection of the small intestine in rats.

General nutrition, intestinal absorption and liver structure and function were compared in rats subjected to: 1) 90 per cent resection of the small intestine, 2) 90 per cent small intestinal bypass with end-to-side jejunoileostomy (ES bypass), and 3) 90 per cent small intestinal bypass with end-to-end jejuno-ileostomy and anastomosis between the excluded segment and the colon (E-E bypass). The E-E bypass group showed the highest mortality rate and the lowest body weight. In this group the haemoglobin concentration, the faecal fat excretion, and the liver function parameters were more abnormal than in resected rats. Rats with E-S bypass showed results in between the other two groups. In none of the animals was fatty infiltration or cirrhosis of the liver observed. It is concluded that intestinal bypass in rats has a more deleterious effect than resection, and this seems to be more pronounced when the excluded segment is anastomosed to the colon. Factors that might be responsible for this effect are discussed.

Alanine Transaminase↗

[The function of the small intestine based on hydrogen test data in men who have been exposed to small doses of ionizing radiation in the cleanup of the sequelae of the accident at the Chernobyl Atomic Electric Power Station].

Sixty-seven participants of the elimination of Chernobyl accident sequels were examined. Of these, 30 persons (44.8%) showed normal characteristics of the three small intestinal functions studied: activity of the intestinal enzyme beta-galactosidase, motility and microbial dissemination, 23 (34.3%) deficiency of the intestinal enzyme beta-galactosidase, and 24 (35.8%) bacterial dissemination of the small intestine. In 12 persons (17.9%), the deficiency of beta-galactosidase was associated with dysbiosis of the small intestine. The incidence of hypolactasia did not exceed the characteristic typical on the whole of the population of the Moscow region.

Accidents↗

Site-dependent small intestinal absorption of ranitidine.

The site-dependent, small intestinal absorption characteristics of ranitidine were estimated by the intestinal steady state perfusion technique (triple lumen tubing system) combined with simultaneous measurement of serum concentrations of ranitidine. Ranitidine 150 mg.l-1 was perfused at 10 ml.min-1 for 180 min in different sites of the small intestine between 65-250 cm beyond the teeth. Each of 9 healthy, male volunteers was examined twice, using perfusion sites in different regions of the small intestine to permit intraindividual comparisons. The absorption rates (micrograms.30 cm-1.min-1) calculated from intestinal samples showed distinct site-dependence; the highest rates (medians 160-923 micrograms.30 cm-1.min-1) were found in the most proximal region (duodenojejunal junction), and the most distal perfusion sites (distal jejunum/ileum) showed median rates from 193 to 265 micrograms.30 cm-1.min-1. In both of these regions there was a significant positive correlation between the net intestinal water flux and the movement of ranitidine. Within the mid-jejunum, every subject showed marked secretion of ranitidine into the gut lumen (medians -338 to -124 micrograms.30 cm-1.min-1), and in this region there was no influence of water flux on ranitidine movement. The intraluminal results were confirmed by the corresponding site-dependent areas under the serum concentration-time curves (AUC), which decreased with the distance of the perfusion site from the teeth. After the more distal perfusions individual AUCs amounted to 64-16% of the AUCs obtained after more proximal applications. The results demonstrate the small intestine as the site of a gradient of absorption of ranitidine.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Histochemical characteristics of the goblet cell mucins and their role in defence mechanisms against Nippostrongylus brasiliensis infection in the small intestine of mice.

The histochemical characteristics of the small intestinal goblet cell mucins of mice and their protective role during the expulsion of Nippostrongylus brasiliensis adult worms were examined. Lectin histochemistry revealed that, before infection, strongly positive stainability of Helix pomatia agglutinin (HPA) and Limax flavus agglutinin (LFA) was observed in all of the small intestinal goblet cell mucins of mice. When 'normal' or 'damaged' N. brasiliensis adult worms were implanted separately into the small intestine of euthymic mice, 50% of them were established in the intestine on day one after implantation, but were subsequently expelled within one week in association with HPA and LFA positive goblet cell hyperplasia. On the other hand, in dexamethasone treated euthymic mice, 70% of implanted 'normal' worms were expelled by day 7 and 80% of implanted 'damaged' worms were expelled by day 4 in association with HPA and LFA positive goblet cell mucins but no goblet cell hyperplasia. This is consistent with the fact that, in spite of no increase in goblet cell number in hypothymic nu/nu mice, all of the implanted 'damaged' worms were expelled within day 5 and 80% of implanted 'normal' worms were expelled by day 14 post implantation, possibly associated with strongly expressed terminal GalNAc and sialic acid residues of the small intestinal goblet cell mucins prior to infection.

Animals↗

[The diagnostic significance of the small bowel enema in lesions of the small intestine requiring surgery].

Given its anatomical location the small intestine is the section of the digestive tract most inaccessible to objective clinical and instrumental examination. For this reason, traditional investigation methods provide inadequate data. In the barium meal X-ray, images are superimposed and the barium excessively fragmented due to the tortuosity and length of the intestinal loop. Access problems limit the value of endoscopy. The indications to angiography are fairly restrictive. Ultrasound and CT scans provide no conclusive evidence in this area. The so-called Small Bowel Enema technique based on the direct infusion of an opaque contrast medium after duodenal intubation remains the only way to obtain images with a satisfactory diagnostic value and should be the method of choice in examinations of the small intestine. Introduced in the twenties, the value of the technique was long questioned, mainly because it was difficult to perform correctly and it is only quite recently that several decisive technological innovations have made it highly reliable. The present paper describes its use on a series of patients with suspected pathologies of the small intestine requiring surgical treatment. The aim is to specify the correct indications to this technique as well as its limitations with a view to promoting its appropriate use in routine practice. MATERIALS AND METHODS. The study covers 42 small bowel enemas performed on 29 male and 13 female patients aged 12-85 years. Indications to the enema included chronic abdominal pain in 28.5%, chronic diarrhea in 21.4%, pain+diarrhea in 14%, subocclusion in 16.5%, faecal blood in 11.9%, abdominal pain+diarrhea+intestinal haemorrhage in 4.7%, anaemia in 2.4%. We adopted the Sellink technique involving intubation with a French 14 Bilbao-Dotter probe and the infusion of 200-350 cc barium sulphate in a 90%-70% P/V concentration at a speed of 100 ml/min, followed by the infusion of 1000-1500 cc 0.5-1% methylcellulose in a 50-50 mixture with water for double contrast purposes. Mean duration of procedure 35-40 mins. RESULTS. We found lesions of the small intestine in 18 cases or 45% (true positives) and no alterations in 18 or 42.8% (true negatives). In 3 cases (7%) the examination was inconclusive. One false positive and one false negative complete the series, making a diagnostic accuracy of 89.1%. In 23.8% of the total series the diagnosis of Crohn's disease was confirmed; cancerous lesions were found in 11.9% and adhesions following earlier surgery in 9.7%. Surgery confirmed this diagnosis in 10 cases (5 tumours, 3 stenoses caused by adhesion fragments, 2 Crohn's disease) and disproved it in one case. Biopsy was used, for objective confirmation in 5 cases (2 Crohn's disease, 3 negatives). In the other cases reliance was placed on clinical course and the patients' response to treatment. DISCUSSION AND CONCLUSIONS. Examination of our results reveals that in most clinical cases the small bowel enema was used to confirm suspected Crohn's disease (31 cases out of 42) and with a very high degree of diagnostic accuracy (96.7%). In line with data in the literature we decided to base the indication to this examination on certain primary symptoms such as chronic abdominal pain, diarrhea, blood in the faeces, whether in isolation or in combination. On this rather general basis the diagnosis of Crohn's disease concerned only 23.8% of the cases studied, an incidence that rose dramatically in the few cases in which the indications were made more specific. We therefore conclude that the adoption of more restrictive criteria will improve the cost-benefit ratio for this test. Another major pathology in which small bowel enema possesses a high diagnostic value is partial obstruction of the small intestine.(ABSTRACT TRUNCATED AT 400 WORDS)

Adolescent↗

Epithelioid cell cultures from rat small intestine. Characterization by morphologic and immunologic criteria.

Rat small intestinal epithelial cell lines have been established in vitro and subcultured serially for periods up to 6 mo. These cells have an epithelioid morphology, grow as monolayers of closely opposed polygonal cells, and during the logarithmic phase of growth have a population doubling time of 19--22 h. Ultrastructural studies revealed the presence of microvilli, tight junctions, an extensive Golgi complex, and the presence of extracellular amorphous material similar in appearance to isolated basement membrane. These cells exhibit a number of features characteristic of normal cells in culture; namely, a normal rat diploid karyotype, strong density inhibition of growth, lack of growth in soft agar, and a low plating efficiency when seeded at low density. They did not produce tumors when injected in syngeneic animals. Immunochemical studies were performed to determine their origin using antisera prepared against rat small intestinal crypt cell plasma membrane, brush border membrane of villus cells and isolated sucrase-isomaltase complex. Antigenic determinants specific for small intestinal epithelial (crypt and villus) cells were demonstrated on the surface of the epithelioid cells, but they lacked immunological determinants specific for differentiated villus cells. An antiserum specifically staining extracellular material surrounding the cells cultured in vitro demonstrated cross-reactivity to basement membrane in rat intestinal frozen sections. It is concluded that the cultured epithelioid cells have features of undifferentiated small intestinal crypt cells.

Animals↗

Intestinal absorption of alpha-methyldopa: in vitro mechanistic studies in rat small intestinal segments.

The mechanism of intestinal uptake of alpha-methyldopa (Aldomet) was investigated using isolated segments of rat small intestine. Incubations were limited to 2 to 5 min as histological examination of the tissue showed significant loss of structural integrity after 10 to 20 min at 37 degrees C. alpha-Methyldopa in the tissue was extracted and assayed by high-pressure liquid chromatography. Corrections were applied for uptake into extracellular spaces using inulin (14C). Uptake was temperature- and concentration-dependent (Km congruent to 10 mM), was dependent on the location in the small intestine and was inhibited by ouabain (2 mM) or lack of sodium or glucose in the incubation medium. alpha-Methyldopa uptake also was inhibited by other neutral amino acids. The mucosal cell layer accounted for approximately 50% of the total drug accumulated in the tissue. Uptake was less when the serosal surface was exposed than when the tissue was everted. The similarity in uptake parameters between alpha-methyldopa and L-phenylalanine (parallel experiments) suggests that alpha-methyldopa is principally absorbed into rat small intestinal mucosal cells via an amino acid transport system.

Amino Acids↗

Effect of small intestinal bypass on hepatic lipid accumulation in rats.

Bypass of large segments of the small intestine to produce weight loss in morbidly obese human subjects has been associated with increased lipid accumulation in the liver. Congenital and nutritional obese rat models were employed to evaluate hepatic lipid accumulation following bypass of 90% of the small intestine. When small intestinal bypass was performed on nutritionally obese rats ingesting a high-fat diet (17%) supplemented with choline dihydrogen citrate (3,4 g/kg diet), a 59% loss of weight at 1 mo was unassociated with increased hepatic lipid accumulation when compared to sham-operated controls. When small intestinal bypass was performed on a nutritionally obese rat fed a relative choline-methionine-deficient diet following intestinal bypass, marked increases in total hepatic lipid and hepatic triglyceride content were evident when compared to the lipid content in sham-operated controls ingesting a similar choline-deficient diet. Hepatic steatosis following small intestinal bypass could be produced in a rat model only by a deficiency in a required substrate for triglyceride conversion to phospholipid and suggests that similar deficiencies may contribute to the increased hepatic lipid accumulation occurring in human subjects following intestinal bypass.

Acetyl-CoA Carboxylase↗

The site of absorption in the small intestine determines diltiazem bioavailability in the rabbit.

PURPOSE: Since the ability of the small intestine to biotransform a drug may decrease in distal segments of the intestine, this study aimed to assess whether the site of administration in the small intestine could affect the systemic bioavailability of diltiazem and its two active metabolites, N-desmethyldiltiazem (MA) and desacetyl-diltiazem (M1). METHODS: Five mg/kg of diltiazem were administered into the lumen of the proximal (0-30 cm, n = 9) or the distal (150-180 cm) small intestine (n = 7) of anesthetized New Zealand rabbits. Blood samples were drawn from the femoral artery for 6 hours, and diltiazem, MA and M1 were assayed by HPLC. RESULTS: The area under the curve (AUC0-->infinity) of diltiazem administered into the distal small intestine was larger than that estimated when diltiazem was given in the proximal segment (14.20 +/- 2.82 vs 8.14 +/- 0.88 micrograms.min/ml, p < 0.05), due to a lower diltiazem oral clearance (440 +/- 78 vs 660 +/- 55 ml/min/kg, p < 0.05). The AUC0-->360 of MA was not affected by the site of diltiazem administration, but the AUC0-->360 of M1 was increased when diltiazem was administered in the distal segment of the small intestine. When administered into the distal segment of the intestine, the molar sum of diltiazem and its active metabolites was 48% greater than when delivered into the 0-30 cm segment of the small intestine; as a consequence, absorption of diltiazem in distal segments of the small intestine may enhance its pharmacological response. CONCLUSIONS: The site of absorption into the intestine modulates the bioavailability of diltiazem and its two active metabolites.

Animals↗

Epicatechin is the primary bioavailable form of the procyanidin dimers B2 and B5 after transfer across the small intestine.

Perfusion of isolated small intestine with the procyanidin dimers B2 and B5 extracted from cocoa indicated that both forms of dimer are transferred to the serosal side of enterocytes but only to a very small extent (<1% of the total transferred flavanol-like compounds). However, perfusion of dimer mainly resulted in large amounts of unmetabolised/unconjugated epicatechin monomer being detected on the serosal side (95.8%). The cleavage of dimer during transfer seemed to be energy-dependent, requiring an intact cell system, as incubation with jejunal homogenates failed to yield epicatechin. Low levels methylated dimer were also detected (3.2%), but no conjugates and metabolites of epicatechin indicating that metabolism of monomer and dimer is limited during dimer cleavage/translocation. The methylation of dimer may be by catechol-O-methyltransferase, however, at high concentrations of dimer COMT activity is reduced leading to an inhibition of both monomer and dimer O-methylation.

Animals↗

Total antioxidant capacity following extrinsic denervation and small intestinal transplantation in the rat.

Transplantation of small intestine in a rat model has been shown to affect expression of neurochemicals within enteric inhibitory nerves. However, the mechanism for altered expression of inhibitory neurochemicals is uncertain. Based on our previous studies, we hypothesized that small intestinal transplantation would result in altered intestinal levels of antioxidant capacity. Glutathione, total antioxidant capacity, and lipid peroxide levels were measured at 3 months following (1) transection of rat small intestine, (2) transection and extrinsic denervation of rat intestine, and (3) isotransplantation of rat ileum or (4) allotransplantation of rat ileum with cyclosporine therapy to suppress rejection. Glutathione levels were not significantly different among the four groups. There were trends toward increased lipid peroxide levels following isografting and extrinsic denervation. Total antioxidant capacity was increased following extrinsic denervation (P=0.05). Increased intestinal total antioxidant capacity in response to extrinsic denervation may represent a compensatory mechanism for protection against oxidative stress. This result enhances our understanding of the relationship between tissue antioxidant levels and alteration of enteric nerves.

Animals↗

Application of pulsed Doppler ultrasound for the evaluation of small intestinal motility in dogs.

The purpose of this study was to verify whether small intestinal peristalsis could be observed and quantitatively assessed using pulsed-Doppler ultrasound. Pulsed-Doppler ultrasound was used to evaluate small intestinal peristalsis after a meal in ten normal dogs and ten sedated dogs. The small intestinal peristalses were measured 0, 1, 3, 6, 9, 12, and 24 hours after a 24-hour fast and after feeding. The number of small intestinal peristalsis were 0.133/min, 0.100/min, 0.033/min, 0.167/min, 0.070/min, 0.067/min, and 0.100/min in the fasted dogs, and 1.667/ min, 0.933/min, 1.133/min, 1.234/min, 1.933/min, 1.533/ min, and 0.533/min in fed dogs, respectively. In the dogs sedated with xylazine HCl, the number of small intestinal peristalsis was significantly reduced (p<0.01). However, in the dogs treated with ketamine HCl and acepromazine, the number of small intestinal peristalsis remained unchanged. Therefore, it can be concluded that pulsed-Doppler ultrasound allows graphic visualization of the intestinal movements, which can be subjected to qualitative and quantitative analysis, and may be suitable for a non-invasive study of small intestinal motility.

Acepromazine↗