Children with screening-detected coeliac disease show increased levels of nitric oxide products in urine.

AIM: Increased concentration of nitric oxide (NO) metabolites, nitrite and nitrate, in the urine is a strong indication of ongoing small intestinal inflammation, which is a hallmark of the enteropathy of coeliac disease (CD). It has previously been shown that children with symptomatic, untreated CD have increased levels of NO oxidation products in their urine. The aim of this study was to investigate whether screening-detected, asymptomatic coeliac children display the same urinary nitrite/nitrate pattern. METHODS: In a multicenter screening study, serum samples were collected from 7208 12-year-old children without previously diagnosed CD. Sera were analysed for anti-human tissue transglutaminase (tTG) of isotype IgA. Small bowel biopsy was performed in antibody-positive children, yielding 153 new cases of CD. In the screening-detected individuals, the sum of nitrite and nitrate concentrations in the urine was analysed and used as an indicator of NO production. For comparison, 73 children with untreated, symptomatic CD were studied. RESULTS: The nitrite/nitrate levels in children with screening-detected CD and those with untreated symptomatic CD did not differ significantly. Both groups had significantly increased urinary nitrite/nitrate concentrations compared to the children with normal small bowel biopsy (p < 0.001). CONCLUSION: Children with screening-detected CD have increased production of NO just as children with untreated symptomatic CD. High NO metabolite levels in the urine may indicate a pathogenetic feature of CD and be a marker of major clinical importance.

Duodenal endocrine cells in mice with particular regard to age-induced changes.

Duodenal endocrine cell types in four age groups of NMRI mice (1, 3, 12 and 24 months old) were identified by immunocytochemistry and quantified by computerized image analysis. Whereas the number of secretin-immunoreactive cells was significantly increased in the 24-month-old group, the number of GIP-immunoreactive cells was reduced in 12-month-old compared with 3-month-old mice. The number of somatostatin-immunoreactive cells was fewer in both the 12- and 24-month-olds vis-à-vis the 3-month-old mice. Whereas serotonin-immunoreactive cells were fewer in both 1-month-old and 12-month-old mice, they were more numerous in 24-month-old mice then in the 3-month-old ones. The number of gastrin/CCK-immunoreactive cells was unaffected by age. The cell secretory index (CSI) of secretin- and serotonin-immunoreactive cells was increased in the 24-month-old mice vis-à-vis the 3-month-old ones and the CSI of GIP- and somatostatin-immunoreactive cells was increased in 12-month-old mice vis-à-vis 3-month-old mice. In contrast, the CSI of somatostatin- and serotonin-immunoreactive cells in 1-month-old mice was lower than that of 3-month-old-mice. The nuclear volume of secretin-, GIP-, gastrin/CCK- and serotonin-immunoreactive cells was less in 1-month-olds than in 3-month-old mice. Whereas the nuclear volume of somatostatin-immunoreactive cells was decreased in 12-month-old animals, that of gastrin/CCK- and serotonin-immunoreactive cells was greater in 24-month-old mice than in 3-month-old ones. It is concluded that these changes may be secondary to structural and functional changes in the gastrointestinal tract caused by ageing. It is possible that these changes are involved in the development of dysfunction of the gut observed at advanced age.

Interruption of early pregnancy with mifepristone in combination with gemeprost.

BACKGROUND: Mifepristone in combination with prostaglandin has been used since 1988 for induction of early abortion. The aim of the present investigation was to assess the tolerance and efficacy of 600 mg. mifepristone orally followed by gemeprost 1 mg. vaginally either 24 hours (group one) or 48 hours (group two) later. METHODS: Sixty-four healthy women applying for abortion within the first 8 weeks of pregnancy were randomly allocated to one of the two treatment groups. Intrauterine pregnancy and gestational age were verified by ultrasonography. Symptoms after administration of mifepristone and gemeprost were recorded, and the patients observed at the hospital for at least three hours after prostaglandin-insertion. Blood samples for blood group, hemoglobin, beta-chorion-gonadotrophin, aspartate-aminotransferase and creatinine were drawn. RESULTS: Outcome was established by gynecological examination, the level of beta-hCG and ultrasonography, at visits one, two and if necessary three to four weeks later. Surgical curettage was performed in case of incomplete abortion, of which there were four in the 24-hour interval group and five in the 48-hour interval group with a success rate (complete abortion) of 55 out of 64 patients (86%). CONCLUSIONS: There was no difference in efficacy or side effects whether the prostaglandin was administered 24 or 48 hours after mifepristone intake, which suggests that the treatment period can be reduced from the conventional 48 hours.

Human rectal endocrine cells and aging.

Endocrine cells of the human rectum were investigated by immunocytochemistry and quantified by computerized image analysis in three different age groups. The age intervals were 20-29, 40-49 and 60-69 years. No statistically significant differences were found between the age groups, regarding the numbers of all endocrine cell types investigated, namely peptide YY (PYY)-, pancreatic polypeptide (PP)-, enteroglucagon-, somatostatin- and serotonin-immunoreactive cells. Nor was there any difference regarding the cell secretory index. Nuclear volume was significantly greater in the 40-49 year olds than the other age groups. There was no statistically significant difference between females and males regarding numbers of the endocrine cell types investigated. It is concluded that age does not affect the endocrine cells of the human large intestine as it was earlier found in animal models of aging. It is imperative that caution should be taken when applying results obtained in animal models of aging in humans.

Ageing and endocrine cells of human duodenum.

Motility and secretory disorders of the gastrointestinal tract and associated glands increase with ageing. The duodenum contains several peptide/amine producing cells that play an important role in regulating gastrointestinal motility and secretion. The present study was performed to elucidate changes in these cells that may have arisen as a result of ageing. A total of four age groups of subjects, aged 1-2, 20-29, 40-49 and 60-69 years were studied. The various endocrine cell types were identified by immunohistochemistry and quantified by computerized image analysis, and two parameters were determined; the number of cells/mm3 epithelial cells and the cell secretory index (CSI), which indicates the immunoreactive secretory granule content of the endocrine cells. Chromogranin A- and serotonin-immunoreactive (IR) cells were fewer in 1-2-year-olds than in 20-29-year-olds. Gastrin/CCK-IR cells were significantly more numerous in 1-2-year-olds and 60-69 years-olds than in 20-29-year-olds. Somatostatin-IR cells were more numerous in the 40-49-year-olds than in the 20-29 years-olds. The CSI was higher in chromogranin A-, gastric inhibitory polypeptide (GIP)-, somatostatin- and gastrin/CCK-IR cells in 1-2-year-olds than in 20-29-year-olds. There was no significant sex difference regarding the numbers and CSI of other endocrine cell types. This study established the absence of sex-related differences in all endocrine cell types investigated, regarding numbers and physiological activity. Age, on the other hand, was shown to be associated with changes in the numbers of CCK-, somatostatin- and serotonin-IR, which may have some bearing on the gastrointestinal disorders of the elderly.

Age-induced changes in the enteric nervous system in the mouse.

The enteric nervous system of the murine gut was investigated by immunocytochemistry in 1-, 3-, 12- and 24-month-old mice, using protein gene product 9.5, a general marker for nerve elements. Myenteric and submucosal plexi were quantified by computerized image analysis. In antrum, there were significantly fewer neurones per ganglion in both myenteric and submucosal ganglia of 12- and 24-month-old mice than in 3-month-old animals. The same was true of duodenum and colon. The relative volume density of nerve fibres in antral muscularis propria was significantly greater in the 1-, 12- and 24-month-old mice than in the 3-month-old mice. In colon, there were fewer submucosal ganglia per millimetre baseline in 1-month-old mice than in 3-month-old mice. The colonic myenteric ganglion in 1-, 12- and 24-month-old mice was smaller than in 3-month-old mice. There was no statistical difference between females and males regarding the number of ganglia per millimetre baseline, ganglionic area, number of neurones per ganglion or the relative volume density of nerve fibres in either the myenteric or submucosal plexi. As the enteric nervous system is responsible for coordinating and integrating the motility of the gut, the ageing-related changes reported here may well be of some relevance for the increased gastrointestinal motility dysfunction in the elderly persons.

Age-related changes in antral endocrine cells in mice.

Antral endocrine cells in four age groups of mice, namely prepubertal (1 month old), young (3 months old), ageing (12 months old) and senescent (24 months old), were detected by immunocytochemistry and quantified by computerized image analysis. A statistical difference was detected between the different age groups regarding the numbers of gastrin-, somatostatin-, and serotonin-immunoreactive cells. The number of gastrin-immunoreactive cells significantly increased between 1 and 12 months, whereas they became significantly fewer between 12 and 24 months. Somatostatin-immunoreactive cell number increased significantly in 1-, 12- and 24-month-old mice, compared with young mice (3 months old). The number of serotonin-immunoreactive cells also increased significantly in 1- and 12-month-old mice as compared with young mice. There was a statistical difference between different age-groups regarding the cell secretory index (CSI) of somatostatin- and gastrin-immunoreactive cells, the CSI of both somatostatin- and serotonin-immunoreactive cells increased significantly in 1-, 12-, and 24-month-old mice, compared with young mice. There was no statistical difference between the different age-groups regarding the CSI of gastrin-immunoreactive cells, nor between males and females regarding the number and CSI of all the endocrine cell types investigated. It is suggested that the large number of somatostatin-immunoreactive cells in ageing and senescent mice might have an impact on the gastric delay seen in the elderly. It was concluded also that the changes in the antral endocrine cells could be involved in the development of dysfunction of the gastrointestinal tract inherent in ageing, or could be secondary to structural and functional changes in the alimentary tract caused by ageing.

How age changes the content of neuroendocrine peptides in the murine gastrointestinal tract.

BACKGROUND: Motility disorders in the gastrointestinal tract increase with ageing, and colorectal carcinoma is a tumor of the middle-aged and elderly. Gastrointestinal secretion, absorption, motility, cell proliferation, local immune defense and blood flow are all regulated by the neuroendocrine peptides. It is conceivable that gastrointestinal disorders at an advanced age may be accompanied by changes in this regulatory system. OBJECTIVE: To ascertain possible age-related changes in neuroendocrine peptides in a rodent animal model. METHODS: The concentrations of various neuroendocrine peptides were determined by radioimmunoassays in tissue extracts from the antrum, duodenum and colon of mice in four different age groups: 1, 3, 12 and 24 months. The neuroendocrine peptides investigated were: secretin, gastric inhibitory polypeptide (GIP), gastrin, motilin, peptide YY (PYY), somatostatin, vasoactive intestinal polypeptide (VIP), substance P, neuropeptide Y (NPY), galanin and neurotensin. RESULTS: Antrum: Concentrations of somatostatin, VIP and substance P decreased significantly in 1-month-old mice (but that of neurotensin increased) compared with 3-month-old mice. In 12-month-old and 24-month-old mice, concentrations of gastrin, somatostatin, VIP, substance P, NPY, galanin and neurotensin all decreased vis-à-vis 3-month-old mice. Duodenum: Whereas the levels of secretin, GIP and neurotensin increased, those of gastrin, motilin, somatostatin, VIP, substance P, NPY and galanin decreased in 1-month-old mice vis-à-vis 3-month-old mice. In both 12-month-old and 24-month-old mice, the concentrations of secretin and GIP increased, compared with those of 3-month-old mice. The levels of gastrin, motilin, somatostatin, VIP, NPY and galanin decreased in both 12-month-old and 24-month-old mice vis-à-vis 3-month-old mice. Substance P and neurotensin concentrations decreased in 12-month-old mice, but not in 24-month-old mice. Colon: In 1-month-old mice the levels of PYY, somatostatin, VIP, substance P and galanin decreased vis-à-vis 3-month-old mice. In 12-month-old mice, the concentrations of PYY, somatostatin, VIP, NPY, galanin and neurotensin decreased compared with those in 3-month-old mice. In 24-month-old mice, the VIP level decreased, whereas the substance P level increased. CONCLUSION: The changes in neuroendocrine peptides observed in the gastrointestinal tract of this murine animal model could be of some relevance for the increased gastrointestinal dysfunction in the elderly human. They may also be involved in the development of colorectal cancer.

Effect of ageing on colonic endocrine cell population in mouse.

BACKGROUND: Dysmotility in the gastrointestinal tract increases with age. Colonic endocrine cells play an important role in regulating intestinal secretion and motility. The objective was to study possible age-related changes in the colonic endocrine cells of an animal model. METHODS: The colonic endocrine cells in four different age groups of mice were investigated by immunocytochemistry and quantified by computerized image analysis. The ages of these groups were 1, 3, 12 and 24 months old. RESULTS: The numbers of peptide YY (PYY), enteroglucagon and serotonin immunoreactive (IR) cells in 1-month-old mice were significantly increased compared with those of 3-month-old mice. Similarly, the numbers of these cells were significantly greater in 12- and 24-month-old mice than in 3-month-old mice. The cell secretory index (CSI) of enteroglucagon and serotonin IR cells was higher in 1-, 12- and 24-month-old mice than in 3-month-old mice. There was no significant difference between the different age groups regarding the CSI of PYY IR cells, nor was there any statistical difference between females and males in all endocrine cell types regarding numbers and CSI. CONCLUSION: It is suggested that the increase in colonic endocrine cells prior to puberty might reflect the role of gut hormones in the development of the gastrointestinal tract. It is speculated further that the increase in colonic endocrine cells with ageing may compensate for increased receptor resistance and/or weakened response of effector organs. It is suggested that the increase in numbers and unchanged CSI of PYY cells with advancing age may be responsible for the slow colonic transit and constipation, both of which increase with age.

Application of computer image analysis in endocrine cell quantification.

Computer image analysis was applied for quantifying endocrine cells by using an automatic standard sequence analysis operation. Two parameters were used, namely the number of cells per mm3 of epithelial cells and the cell secretory index (the volume of the immunoreactive secretory granules per cell). The first indicates the variation in the anatomical peptide-producing unit and the second the synthesis and secretion activity of the cell. The endocrine cells chosen in this study were chromogranin-immunoreactive cells and secretin-immunoreactive cells in the human duodenum. The measurements were made by five different investigators with different backgrounds in order to evaluate the effect of the intra- and inter-individual variation. This study showed that the intra- and inter-individual variation had no impact on the results. Comparisons with the classical point-counting method considered to be easy and most efficient in volumetry showed that the present approach is between two and three times faster and less strenuous for the performer. It is concluded that this approach seems to be suitable for adaptation in morphometric studies when information is required about the changes in the number of endocrine cells and about changes in secretory activities.

Evaluation of immunohistochemical staining of human duodenal endocrine cells after microwave antigen retrieval.

The effect of microwave antigen retrieval on the immunostaining of human duodenal endocrine cells in formaldehyde-fixed, paraffin-embedded material was investigated. The sections were immunostained by the avidin-biotin complex (ABC) and immunogold-silver autometallography (IGSS) methods with and without prior microwave treatment. Dilutions of up to 1:30,000 of the following antisera/antibodies were used: anti-chromogranin A, anti-chromogranin AB, anti-secretin, anti-gastrin, anti-gastric inhibitory polypeptide, anti-somatostatin and anti-serotonin. The detection threshold for all the antibodies was lower after antigen retrieval, and the primary antibody could be used in higher dilutions. The dilutions varied for different antibodies and were between two and ten times the optimal dilution without antigen retrieval. At extremely high dilutions of, or without, the primary antibody, non-specific staining of some lymphocytes and the mucus of some goblet cells was observed when the avidin method was applied, but not with the immunogold technique. This phenomenon was not observed when optimal dilution or a lower dilution was used. This seems to have been caused by the binding of the avidin-biotin complex to epitopes in these structures unmasked by microwave treatment when competition with specific binding sites was absent.