Prognostic significance of standardized AgNOR analysis and Ki-67 immunostaining in gastrointestinal stromal tumors

OBJECTIVE: To evaluate the prognostic utility of argyrophilic nucleolar organizer region (AgNOR) protein parameters and Ki-67-immunostained growth fraction (Ki-67 labelling index) and to correlate AgNORs with Ki-67 LI and the main clinicopathologic parameters in gastrointestinal stromal tumors (GISTs). STUDY DESIGN: On 55 patients with surgically excised GISTs, visualization and quantification of AgNORs were performed as specified in the guidelines of the Committee on AgNOR Quantification. RESULTS: AgNOR protein area (NORA) > or = 5.28 microns 2 was statistically associated with mitotic rate > or = 5 x 10 high-power fields (hpfs) (P < .001) and presence of necrosis (P < .001); Ki-67 LI > or = 9.69% was significantly associated with mitotic rate > or = 5 x 10 hpfs (P < .001), size > or = 5 cm (P = .033) and presence of necrosis (P < .001). Ki-67 LI and NORA strongly correlated. Preliminary Kaplan-Meier survival analysis showed that an increased value of NORA, Ki-67 LI, mitotic rate, tumor size and presence of necrosis had a negative influence on patient survival. Multivariate regression analysis showed that only NORA and Ki-67 LI were independent parameters in predicting the clinical outcome for patients with GISTs. Mitotic rate and necrosis remained as independent prognostic factors when NORA and Ki-67 LI were not allowed to enter in models. CONCLUSION: AgNOR protein quantity, as determined by image cytometry, and Ki-67 immunostaining seem to represent reliable predictive parameters in GISTs and are independent of mitotic rate, tumor dimension and necrosis.     

Iron absorption by small intestine of chickens

Iron (Fe) absorption by three segments (duodenum, jejunum, and ileum) of the small intestine of chickens was studied by a perfusion technique in vivo in closed circuit using⁵⁹Fe Cl₃ and was related to the histological characteristics of each segment. The serosal transfers of Fe for the duodenum and jejunum were the same (14%/cm), but significantly different (p<0.05) from those of the ileum (9%/cm), which may be explained by the morphological and histological properties of the gut of chickens. However, the presence of Fe in blood and in liver was significantly lower after perfusion of the jejunum and ileum than after perfusion of the duodenum. It is concluded that chickens show an early adaptation of small intestine to Fe absorption in response to the considerable loss of Fe suffered during the laying process.     

Localized colonic inflammation increases cytokine levels in distant small intestinal segments in the rat

Local inflammation in the colon has been associated with nutrient malabsorption and altered motility in the small bowel. These remote effects suggest the release of mediators which can act (or alter) the function of intestinal segments located far from the primary area of inflammation. This study describes the changes in the expression of pro-inflammatory cytokines in the colon and in various segments of the small intestine in two rat models of experimental colitis. Colitis was induced by the intracolonic administration of 100 microL of 6% iodoacetamide or 250 microL of 2, 4, 6-trinitrobenzene sulfonic acid. Levels of interleukin one beta, interleukin 6, and tumor necrosis factor alpha were measured by ELISA in tissue homogenate sampled from duodenum, jejunum, ileum and colon at different time intervals. In homogenates of strips isolated from duodenum, jejunum and ileum, tumor necrosis alpha and interleukin-6, increased significantly 3-6 h after iodoacetamide or TNBS administration and remained elevated until the colonic inflammation subsided. Interleukin one beta showed comparable but delayed increase. Similar, but more pronounced increase of the three cytokines was noticed in areas of the colon adjacent to the ulcer. Histologic examinations revealed important inflammatory changes in the colon; however, examination of sections from the small intestines did not reveal significant differences between controls and rats with colitis. In conclusion, expression of pro-inflammatory cytokines is increased in remote segments of the small intestines during colitis. The findings may provide a partial explanation or a molecular substrate for the associated small bowel dysfunction.     

Somatostatin binding sites in cytosolic fraction of rabbit intestinal mucosa: distribution throughout the intestinal tract

Specific binding sites for somatostatin have been identified in cytosolic fraction of both small and large intestinal mucosa. The stoichiometric data suggested the presence of two classes of binding sites in each part of the intestine. The binding capacity varied depending on the segment considered (rectum greater than duodenum = jejunum greater than ileum, caecum and colon). However, the affinities of the binding sites were similar throughout the whole intestinal mucosa, with the exception of rectum which showed higher Kd values. The binding sites were shown to be highly specific for somatostatin since neuropeptides such as vasoactive intestinal peptide, neurotensin, substance P and Leu-enkephalin did not show any effect upon somatostatin binding.     

Mechanism of the serotonin effect on motility of the duodenum,ileum, and jejunum in awake rabbits

I. v. administration of serotonin to alert rabbits produced a phasic change of contractile activity of duodenum, ileum, and jejunum including excitatory and inhibitory components. It is shown that stimulation of the small bowel motility is caused by serotonin activation of non-cholinergic excitatory mechanism with participation of effector cholinergic neurones. The initial suppression of the motility is caused by participation of nonadrenergic noncholinergic inhibitory mechanism, and the secondary inhibition of contractile activity of a small bowel with serotonin has an adrenergic nature.     

Regional differences in the appearance of adult-type glycosphingolipids in the small intestine of inbred rats at weaning time

The small intestine of 15- to 33-day-old rats was cut into four segments: duodenum, proximal jejunum, distal jejunum, and ileum. Neutral glycosphingolipids and gangliosides were purified from each segment and analyzed by thin-layer chromatography in order to study the developmental appearance of adult-type glycolipids at each level of the small intestine. Type 1 A-6 glycolipid was first detected in the ileum at 15 days and subsequently in the jejunum and duodenum at 19 days of age. N-Glycolylneuraminic acid was expressed first in the ileum at 17 days, then in the proximal jejunum at 21 days, but only after 29 days in the duodenum. In each region, 6-8 days were required between first detection and full expression of N-glycolylneuraminic acid. The presence of 2-hydroxylated fatty acids in glucosylceramide was found first in the ileum at 19 days, 2-3 days before appearing in the duodenum and proximal jejunum. A period of 2-3 days was necessary to reach full adult-type level of 2-hydroxylated fatty acids in glucosylceramide. These results show that adult-type glycolipids appear earlier in the distal than in the proximal region of the rat small intestine, and that different glycolipids appear at different times and at different rates. The finding that the biochemical differentiation of the whole small intestine expands over a period of 3 days to 2 weeks, depending on the region and the glycolipid, before being fully completed indicates that, in addition to the time lag observed between the distal and the proximal region, the new cells arising from the crypt of Lieberkhün after 15 days of age are not at once fully differentiated.     

The morphometry and biomechanical properties of the rat small intestine after systemic treatment with epidermal growth factor

Morphometric and passive biomechanical properties were studied in isolated segments of the duodenum, jejunum and ileum in 22 EGF-treated rats and 12 control rats. The rats were allocated to groups with EGF treatment for 2, 4, 7, and 14 days (n = 6 for each EGF treatment group except n = 4 for the 14 days group) or saline treatment (n = 3 for each group). The intestinal segments were pressurized with Krebs solution from 0 to 8 cmH2O for duodenum and 0 to 6 cmH2O for jejunum and ileum using a ramp distension protocol. The diameter and length were recorded at different pressure levels. Circumferential and longitudinal stresses (force per area) and strains (deformation) were computed from the length, diameter, pressure and the zero-stress state data. EGF treatment was associated with pronounced morphometric changes, e.g., the wall thickness, wall area, and the circumferential lengths significantly increased during EGF treatment in all intestinal segments (P < 0.05). Histological analysis showed that the thickness and area of the layers increased after EGF treatment. With respect to the biomechanical data, the opening angle increased in all segments during EGF treatment with the highest value in the 14 days EGF treatment group (P < 0.05). The same result was found for residual strain and the residual strain gradient through the intestinal wall. Linear regression analysis demonstrated that the opening angle mainly depended on the mucosa thickness and area. Furthermore, the circumferential stiffness increased in the duodenum and decreased in the jejunum and ileum during EGF treatment. A plateau was reached after 7 days where after it started to normalize (P < 0.01). In the longitudinal direction, all intestinal segments became stiffer after EGF treatment for 7 days. After 14 days the curve started to normalize in duodenum and jejunum but not in the ileum.     

Activity of Peptide Hydrolases in Epithelial and Subepithelial Layers of Small Intestine of Rats of Different Age after Protein Deprivation

A significant decrease of protein content in epithelial, stromal, and muscular-serosal layers of jejunum and ileum, especially in aged animals, is revealed in rats of different age groups (young, mature, aged) after 10-day-long protein deprivation. The responses of peptide hydrolases (aminopeptidase M and glycylleucine dipeptidase) were different. There were, as a rule, no changes of these enzyme activities in small intestine of young rats, except for decreased activity of aminopeptidase M in stromal layer of jejunum and increased activity of both peptide hydrolases in epithelial layer of ileum. In mature animals, increased activities of these enzymes also was observed in the epithelial layer of jejunum and ileum and in the stromal layer of ileum in comparison with the rats receiving a full-value nutrition. In aged rats, a much more pronounced rise of these enzyme activities in the layers of small intestine was revealed after protein starvation in comparison with young and mature rats. Probably, such increase of the peptide hydrolase activities in the layers of small intestine after protein deprivation can be considered as an adaptive-protective reaction of the organism in response to formation of significant amounts of low-molecular peptides as a result of protein catabolism     

Absorption and metabolism of purines by the small intestine of the chicken.

1. Absorption of purines and their metabolism by the small intestine were estimated by using the everted gut sacs from the duodenum, jejunum and ileum of the chicken. 2. When no purine was added to the mucosal fluid, large amounts of uric acid, much less but appreciable adenine, hypoxanthine and xanthine and no detectable guanine were released from both sides of all segments of the small intestine, and these released amounts were largest in the duodenum. 3. Similar absorption rates of adenine from the jejunum and ileum were about 1.7-3.0 times as high as those of hypoxanthine and uric acid from these intestines and those of adenine and uric acid from the duodenum (P less than 0.05). 4. Guanine was not absorbed unchanged from any segments of the intestine and a little xanthine was absorbed only from the jejunum and ileum. 5. Guanine and xanthine seem to be absorbed in uric acid form, hypoxanthine in xanthine and uric acid forms and adenine in hypoxanthine form, from the small intestine especially from the jejunum. 6. Adenine, guanine, xanthine and hypoxanthine were greatly metabolized in the mucosa of the duodenum, and the conversions of hypoxanthine to xanthine and uric acid were most active.     

La TEP des tumeurs neuroendocrines de l’intestin grêle

This article focuses on the indication and the choice of tracer for PET/CT in case of neuroendocrine tumours (NET) of the small intestine, which are the most common digestive NETs. PET/CT can be used to search for the primary NET in case of detection of a metastasis, for staging and determination of resectability, for restaging, for optimising and determining the efficacy of therapeutic modalities in extended or recurrent forms. Currently, three types of PET tracers are routinely used: FDG can be useful in the case of aggressive NET especially the duodenum and proximal jejunum, FDOPA is the best tracer in the case of NET of distal jejunum or of ileum, the labeled somatostatin analogues ⁶⁸Ga in the case of well-differentiated NET from the duodenum or proximal jejunum, or irrespective of the location of primary NET if treatment with somatostatin analogue is intended, to confirm the overexpression of somatostatin receptors by lesions.     

化学药物对家兔离体小肠平滑肌电生理特性的影响

观察各种化学药物对家兔离体小肠各段平滑肌的作用,采用常规离体灌流的十二指肠、空肠及回肠平滑肌标本作舒缩运动实验,记录用药前后各段小肠平滑肌的收缩活动特征及变化规律.结果显示：不同浓度的乙酰胆碱和磷酸组织胺能增强小肠各段平滑肌的收缩频率与幅度,其幅值变化与用药前有显著性差异（P＜0.01） ,并呈剂量依赖性;而不同浓度的肾上腺素和阿托品则抑制小肠各段平滑肌（P＜0.01） .不同肠段对各种化学药物的作用存在着差异,一般十二指肠作用最强,空肠次之,回肠最差.     

Fine needle aspiration of gastrointestinal stromal tumors

OBJECTIVE: Gastrointestinal stromal tumors (GISTs) are uncommon mesenchymal tumors of the gastrointestinal tract. Fine needle aspiration (FNA) is one option for diagnosing GISTs before surgery. This study was designed to evaluate the clinical utility of FNA in the diagnosis of GISTs. STUDY DESIGN: FNAs from 19 GISTs originating in the stomach, small bowel and colon obtained from 1988 to 1998 were studied. Immunocytochemistry was performed on 12 cases. The GISTs were classified as benign, borderline and malignant, according to location, size, mitotic activity and clinical outcome. RESULTS: Benign (three) and borderline (five) GISTs were all spindle cell type; malignant GISTs included five spindle cell type and six epithelioid type. Most smears contained abundant cellular material. Benign and borderline GISTs of spindle cell type tended to have cells arranged in tightly cohesive clusters, while malignant GISTs were more likely to exhibit loosely cohesive groups with many single cells, occasional nuclear pleomorphism, hyperchromasia and irregular nuclear contours. Epithelioid-type GISTs mimicked adenocarcinoma. Mitoses were seldom observed in either type. CD117 (KIT protein product) was demonstrated by immunocytochemistry in 9 cases, CD34 in 11, desmin in 3, S-100 protein in 2 and smooth muscle actin in 6 cases. CONCLUSION: FNA can be used to diagnose GISTs as spindle cell and epithelioid types, but cytomorphology alone cannot be used to assess malignant potential. Immunocytochemical staining for CD117 is helpful in confirming the diagnosis. Care must be taken to differentiate epithelioid-type GISTs from adenocarcinoma.     

Changes in thymidine uptake in the gastrointestinal tract of the rat following treatment with 16,16-dimethyl prostaglandin E2

Thymidine uptake in the organs of the gastrointestinal tract of the rat was studied to determine if cell synthesis was involved in the increases in weight of the stomach, small intestine and colon which result from treatment with 16,16-dimethyl prostaglandin E₂ (16,16-dimethyl PGE₂). Animals were treated for 2 days with 16,16-dimethyl PGE₂. They were injected with the ³H-thymidine, sacrificed and the organs of interest were removed. The total amount of tritium in the stomach, duodenum, jejunum, ileum, and colon was determined.Thymidine uptake was significantly increased in the duodenum (1.50 times), jejunum (1.53 times), and colon (1.40 times) but not in the stomach and ileum. The increases were dose related in the duodenum and jejunum. The colon showed a similar dose response pattern but the changes with dose did not reach significance.These results confirm and extend a previous report that 16,16-dimethyl PGE₂ increased thymidine uptake in the duodenum but not the stomach (1). This is different from gastrin which has been shown by others to increase thymidine uptake in the stomach, duodenum, ileum and colon (2,3).     

Lactic acid is absorbed from the small intestine of sheep

A series of experiments was conducted in vivo on anaesthetized sheep to explore the hypothesis that lactic acid is absorbed from the small intestine of sheep. Test solutions varying in lactic acid concentration, pH, osmolarity, and with fixed physiological concentrations of volatile fatty acids (VFAs), K+, Na+, NH4 +, Cl-, and PO4 (-3), were separately introduced into clean, surgically sealed pouches. Studies were undertaken in 27 sheep, each with three pouches in the middle of the duodenum, jejunum, and ileum. Samples were taken at 15-minute intervals for 60 minutes to determine the absorption rates. The experimental results showed that L- and D-lactic acid were absorbed from the pouches of the duodenum, jejunum, and ileum throughout the 60 minutes. In the test solutions with pH 5.3, 420mOsmol/kg, and 12.5mM lactic acid that are in vivo conditions of light lactic acidosis, the mean absorption rates of D-lactic acid and L-lactic acid pooled from three pouches were similar, 0.07micro mol/cm2/min and 0.06micro mol/cm2/min, respectively, based on absorptive surface area. The mean absorption rates of DL-lactic acid from the duodenum, jejunum, and ileum pouches were almost the same, 0.14, 0.14, and 0.11micro mol/cm2/min, respectively. The absorption of lactic acid varied depending on lactic acid concentration, and there was a curvilinear relationship between lactic acid concentration and its absorption rate. A decrease in pH and osmotic pressure resulted in significant, corresponding increases in the absorption of lactic acid (P<0.0001 and P<0.05, respectively).     

Comparison of Endoscopic and Open Resection for Small Gastric Gastrointestinal Stromal Tumor

The National Comprehensive Cancer Network recommends conservative follow-up for gastric gastrointestinal stromal tumors (GISTs) less than 2 cm. We have previously reported that the mitotic index of 22.22% of small gastric GISTs exceeded 5 per 50 high-power fields and recommended that all small gastric GISTs should be resected once diagnosed. The aim of the present study is to compare the safety and outcomes of endoscopic and open resection of small gastric GISTs. From May 2010 to March 2014, a total of 90 small gastric GIST patients were enrolled in the present study, including 40 patients who underwent surgical resection and 50 patients who underwent endoscopic resection. The clinicopathological characteristics, resection-related factors, and clinical outcomes were recorded and analyzed. The clinicopathological characteristics were comparable between the two groups except for tumor location and DOG-1 expression. Compared with the surgical resection group, the operation time was shorter (P = .000), blood loss was less (P = .000), pain intensity was lower (P < .05), duration of first flatus and defecation was shorter (P < .05), and medical cost of hospitalization was lower (P = .027) in the endoscopic resection group. The complications and postoperative hospital stay were comparable between the two groups. No in situ recurrence or liver metastasis was observed during follow-up. Endoscopic resection of small gastric GISTs is safe and feasible compared with surgical resection, although perforation could not be totally avoided during and after resection. The clinical outcome of endoscopic resection is also favorable.     

Cholera toxin: Radio-iodination and uptake by the intestine of suckling rats

1. The chloramine-T procedure was employed to radio-iodinate cholera toxin using Na¹²⁵I. The procedure was found to be efficient and reproducible.2. Intragastric injections of both the labelled and the unlabelled toxin produced (a) significant increases in intestinal fluid accumulation as measured by the fluid accumulation ratio; (b) significant increases in cAMP levels; and (c) significant decreases in cAMP-phosphodiesterase activities when compared with the controls suggesting that radio-iodination did not impair the biological activity of the toxin.3. In vivo uptake studies of the labelled toxin by different parts of the intestine indicated that the uptake by the duodenum and jejunum was high and rapid when compared with the ileum implying that there are more binding sites (or receptor proteins) for cholera toxin in the duodenum and jejunum than in the ileum.     

Ontogeny of Somatostatin Binding Sites in the Rabbit Small Intestinal Epithelial Tract

Abstract

The ontogenic patterns of somatostatin content and its binding sites were studied in small intestine in the developing rabbit from birth until the adult stage. A peak of somatostatin concentration was observed immediately after birth (day 0) and at day 10 in duodenum and jejunum, followed by a decrease at day 15 and a new and gradual augment thereafter. Ileal somatostatin concentrations decreased after birth up to day 15 and then increased progressively with age until the adult period. The somatostatin binding capacity in cytosol of the mucosal layer of duodenum, jejunum and ileum decreased from birth until 10 days and increased thereafter up to the adult period. However, no age differences in dissociation constants were observed. Interestingly, there was an apparent lack of high-affinity sites immediately after birth (day 0) whereas two classes of binding sites were seen thereafter. These results suggest that somatostatin may be involved in the functional and anatomical development of the small intestine during the neonatal period.     

Effects of met-enkephalin on the mechanical activity and distribution of met-enkephalin-like immunoreactivity in the cat small intestine

Naloxone-dependent effects of Met-enkephalin (10(-8) M) on the spontaneous and electrically induced mechanical activities were studied in longitudinal and circular preparations isolated from the cat duodenum, jejunum and ileum. Met-Enkephalin changed the spontaneous activity of all preparations tested with the exception of the circular preparations from the ileum. Met-Enkephalin-induced responses of the longitudinal preparations from the ileum were abolished by treatment with tetrodotoxin (10(-7) M), while the responses of both longitudinal and circular preparations from the duodenum and jejunum were only partially depressed, being resistant to tetrodotoxin components. The latter were most pronounced in the duodenum. The neurogenic electrically induced (0.5 msec, 5 Hz, 150 pulses) responses of all the preparations consisted mainly of contractile components which were significantly and naloxone-dependently reduced by Met-enkephalin (10(-8) M). The contractile components of the responses, which were reduced by Met-enkephalin, were entirely abolished by atropine (3 x 10(-6) M). Both Met-enkephalin and atropine inhibitory effects on the neurogenic responses were more pronounced in the ileum. Met-Enkephalin was found in nerve fibers of the myenteric plexus distributed mainly among the circular muscle. Single immunoreactive nerve fibers were observed in the longitudinal muscle layer of the duodenum but not in the jejunum and ileum. The distribution of Met-enkephalin-like immunoreactivity along the small intestine did not show significant differences among the three intestinal regions tested. The results obtained suggest that Met-enkephalin can modulate the mechanical activity of the cat small intestine, inhibiting cholinergic transmission and/or activating smooth muscle opioid receptors.(ABSTRACT TRUNCATED AT 250 WORDS)