key: cord-015335-l0kjxhd1 authors: nan title: Irish Society of Gastroenterology: Proceedings for summer meeting – 26th/27th May 1995 in Galway date: 1995 journal: Ir J Med Sci DOI: 10.1007/bf02967835 sha: doc_id: 15335 cord_uid: l0kjxhd1 nan Inflammatory bowel disease (IBD), including Crohn's disease (CD) and ulcerative colitis (UC) are associated with a diverse group of extra gastrointestinal features. However, unlike patients with irritable bowel syndrome (IBS), there have been few reports of gynaecological abnormalies in IBD. The aim of our study was to observe the effect of IBD and IBS on the age of menarche and menopause in female's. 153 subjects (mean age 36 yr, range 19-95 yr), resident in Dublin, participated in the study. These included UC (n=40), CD (n=26), IBS (n=52) patients and 35 healthy female controls. The different subgroups were matched for age and marital status. Information from subjects was obtained by a written questionnaire. Menarche was at a significantly later date (student t test, p value < 0.05) in patients with CD (13.9 + 1.14 yr) and UC (13.5 + 1.4 yr) than in normal controls (12.8 + 1.5 yr). No significant differences in age of menarche were observed between the IBS (13.2 + 1.6 yrs) and control groups or between the CD and UC groups (p value > 0.05). The date of menopause was also significantly early (Mann-Whitney U-test) in patients with CD (41.8 _+ 2.4 yr) and UC (43.4 + 2.1 yr) than in normal controls (46.8 + 3.3 yr). The differences between the IBS group (42.8 + 5.6 yr) and the controls or the CD and UC patients were not statistically significant. Conclusion: The fertile period of females with IBD is significantly reduced compared to healthy controls. This difference is not observed in IBS patients. The underlying mechanism is not known but may involve the abnormal immune responses observed in these patients. Chloride (CI) secretion by the human colon epithelium provides the osmotic driving force for fluid secretion into the colon lumen. Under 'normal' conditions this fluid is readily reabsorbed. In secretory states the volume of fluid secreted secondary to CI secretion may exceed the absorptive capacity of the epithelium and diarrhoea results (secretory diarrhoea). Increase in secretion by the colon epithelium is also thought to play a role in the diarrhoeas which occur in inflammatory disease of the colon. In this study we examined tile pathways for CIsecretion in human colon and some aspects of their cellular control. Normal human colon epithelium was dissected from colectomy specimens, mounted in Ussing chambers and voltageclamped to 0 milli-volts. When necessary a single membrane of the epithelial cell was permeabilised with nystatin in order to study the 'opposite' membrane in isolation. Entry of CI across the basolateral membrane of the epithelial cells was inhibited by basolateral addition of a combination of bumetanide and DIDS. Entry of CI at the basolateral membrane was found to be rate limiting for transceUular CI secretion. This suggests that basolateral CI entry to the cell ocurs via Na*/K § 2C1 cotransport and/or CI/HCO~-exchange. CI ~ diffuses across the cell and is extruded at the apical membrane via CI channels which were inhibited by a combination of anthracene-9carboxylic acid (A-9-C) and 5-nitro-2-(3-phenylpropylamino)benzoic acid NPPB. The NPPB inhibited channel was stimulated by experimental manoeuvres designed to increase intracellular I.J.M.S. ~vc,~.~ wvastroen.ero.06~ July, August. September, 1995 Ca 2.. The A-9-C inhibited channel was stimulated by an increase in cellular cAMP (addition of forskolin). This demonstrates that CI secretion across the apical membrane occurs via AMP and also by Ca 2* activated pathways. Vectorial trans-cellular C I transfer occurs in a basolateral to apical direction. CI" uptake across the basolateral membrane is the rate limiting step in this process and occurs via Na §247 cotransport and/or CI/HCO 3" exchange. These mechanisms of CI" uptake are inhibited by bumetanide and DIDS respectively. CI secretion across the apical membrane occurs by two separate types of CI-channel (which may be active simultaneously), a cAMP stimulated CI" channel which is inhibited by A-9-C, and a Ca 2 § stimulated C I channel which is inhibited by NPPB. The understanding of the transcellular mechanism of CI" secretion provided here provides a basic model for CI (and fluid) secretion in human colon epithelium and allows for study of how these processes are affected by neuro-humoral and pharmacologioal agents. Gastric carcinoma is the most common malignancy in China, the fourth most common in Western.Europe and a major cause of cancer morbidity and mortality worldwide. It is well known that metastases, especially distant organ metastases through the blood vascular system, have been the most important cause of patients' death. So far, markers valuable in predicting the metastatic potential of gastric carcinoma have not been identified and the mechanism of metastasis has been unclear. The aim of this study was to determine whether DNA ploidy analysis had any objective value in predicting the metastatic potential of gastric carcinoma to distant organs. DNA ploidy of 57 gastric carcinomas with metastases (12 liver, 1 adrenal, 4 ovary and 48 lymph node) was measured by flow cytometry. DNA aneuploidy was significantly related to liver metastases: 9 out of 12 gastric carcinomas with liver metastases were aneuploid (75%) compared with 13 out of 45 (28.8%) cases without liver metastases (p<0.01); the one gastric carcinoma with adrenal metastasis was also aneuploid. DNA ploidy was not related to ovarian or lymph node metastases. The results suggest that an aneuploid DNA pattern is a predictor of high risk potential for metastases to the liver and may be a useful tool in the "followup" of patients with gastric carcinoma in detecting those at high risk of developing metastases following surgical resection. Liver transplantation (OLT) is complicated by acute cellular rejection (ACR) in up to 80% of patients with a peak incidence in the first two weeks following surgery. A diagnosis of ACR is suspected on the basis of an acute deterioration in biochemical liver function, confirmed histologically with liver biopsy and associated with recipient T cell activation. To examine a component of the immunological events associated with ACR we have assessed the changes in T cell surface molecules and T cell activation status following OLT. Using flow cytometry a total of twenty four peripheral blood T lymphocyte profiles were analysed in six patients over a four week period following liver transplantation, during which four episodes of histologically confirmed ACR developed in three patients. Blood was stained with monoclonal antibodies against CD3, CD4 and CD8 molecules and with dual labelling of the same markers in combination with HLA-DR to define the activation status of the T lymphocyte subsets. Results: 1. Consistent changes were not observed in the HLA-DR negative CD4+ and CD8+ populations or the ratio of CD4+:CD8+ cells during ACR, a finding which has been equivocal in other studies. 2. All four episodes of ACR were associated with an increase in the activation status (HLA-DR antigen expression) of the CD3+, CD4+ and CDS+ populations for each patient as compared to baseline: Mean % CD3+HLA-DR+, CD4+HLA-DR+ and CD8+HLA-DR+ at baseline and during ACR was 2.8 vs 8.1%, 2.0 vs 12.5% and 1.6 vs 20.8% respectively. 3. Successful treatment of 3 episodes of rejection was accompanied by a decrease in the activation status of the three populations: CD3+, CD4+ and CDS+. Mean % CD3+HLA-DR+, CD4+HLA-DR+ and CD8+HLA-DR+ during ACR and following treatment was 8.1% vs 0.86%, 12.5% vs 0.86% and 20.8% vs 4.8% respectively. In two patients no episodes of ACR developed following transplantation. In neither of the subjects were there consistent changes in the activation status of the T cell subsets. In conclusion, we have identified consistent patterns of change in T lymphocyte markers following liver transplantation related to significant clinical events. Serial measurements of these markers may be of use in evaluating patients following transplantation and may predict those at risk of ACR. We have studied 37 haemophiliacs. None presented with liver related disease and all were contacted by postal questionnaire. 30 had haemophilia A, 3 haemophilia B, 4 Von Willebrand disease (VWD) and one was a carrier. There were 34 males and 3 females, ages ranged from 2 to 68 yr. All had ELISA and recombinant immunoblot (RIBA) testing. 20 tested positive by RIBA and 6 of these have circulating virus, demonstrated by polymerase chain reaction (PCR). 17 tested negative for HCV. Mean ALT in the positive groups was 1.59ukat/L, range 0.22 to 6.36, in those negative mean was 0.48ukat/L, and range 0.23 to 0.89. The positive group were older, (mean age 33 yr, range 18 to 68 yr), than those negative (mean age 20 yr range 2 to 51 yr). Conclusions: HCV has been demonstrated in 54% of this group, which is at the lower end of the reported range. Liver function tests reflected the presence of virus. HCV was commoner in older patients, possibly reflecting increased exposure to factor VIII. Although HCV is a slowly progressive diseases because of the improved life span in haemophiliacs, it is likely to be clinically significant in such patients. Whether duodenitis is a distinct clinical entity on its own or a stage in the evolution or resolution of duodenal ulcer disease is still controversial. The aim of our study was to observe the histology of differnt endoscopic patterns of these lesions. The case records of 41 consecutive patients (M:F ratio 2:1, mean age 38 yr, range 21 -48 yrs) with endoscopic evidence of duodenitis were stud.ied. Patients on anti ulcer medication before endoscopy were excluded from the study. All the endoscopies were performed in a single GI unit. Biopsies for histology were taken from the endoscopically inflamed areas of the bulb. The presence of Helicobacter Pylori (Hp) infection was checked with the deltawest test. 4 different.endoseopic patterns of duodenitis were observed: patchy (n=8, 19%), diffuse (n=16, 39%), erosive (n=ll, 27%), and nodular (n=6, 15%). 12 (30%) had normal histology (4/8 of patchy, 6/16 of diffuse, 1/11 of erosive and 1/6 of nodular duodenitis). In the remaining 29 patients, histology showed acute inflammation in 17 (59%), chronic inflammation in 8 (28%) and gastic metaplasia in 4 (13%). Acute inflammation was most frequently seen in erosive duedenitis (90%), and gastric metaplasia in nodfflar duedenitis (40%). Of the 29 patients with abnormal histology, deltawest test was positive in 20 cases (69%), most frequently in erosive duodenitis (90% of the cases). Conclusion: The endoscopic diagnosis of 'patchy' duodenitis is unreliable and therefore if suspected, should be confirmed on histology. Erosive duodenitis is the type most accurately diagnosed endoscopically, most frequently associated with underlying acute inflammation and the presence of Hp infection. Nodular duodenitis is most commonly associated with underlying gastric metaplasia. Non-invasive tests to detect H.pylori are necessary for epidemiological studies, investigation of dyspepsia and for follow-up after eradication treatment. The 13Carbon urea breath test (UBT) is highly sensitive and specific. However, the current recommended protocol for the UBT, including an overnight fast, the use of large reservoir bags, multiple positional changes and sample collections render the test unsuitable for widespread use. The aim of this study was to assess the sensitivity and specificity of a modified, simple UBT. Subjects undergoing endoscopy for investigation of dyspepsia or after a course of eradication therapy were recruited. Nonfasting patients were administered a standard motility-inhibiting liquid meal (50mls Calogen + 50mls Ensure). Immediately after, breath samples were collected in duplicate in 20ml vaeutainers via straws. Subjects then received 100 mg of carbon urea dissolved in 100 ml of water. After sitting for 30 min, repeat breath samples were collected in vacutainers. An excess of ~3CO2 excretion of 5 per ml was taken as a positive result. The UBT results were compared with antral and corpus histology (x4), antral and corpus culture (x2) and antral CLO test. The "gold standard" was defined by the results of any two of the biopsy based methods. 169 subjects were recruited (98 male), mean age 42 yr (range 17-19). 89 subjects had a positive UBT; 85 true positives and 4 false positives. 80 subjects had a negative UBT; 78 true negatives and 2 false negatives. Therefore the sensitivity was 97.7% and the specificity was 95.1%. Conclusion: This modified UBT is user-friendly, ideal for use in a primary care setting and is as reliable as other methods for the non-invasive detection of H.pylori. U.C.C. High oestrogen states (pregnancy, the use of the oral contraceptive pill, phases of the menstrual cycle) are associated with hypertension and oedema secondary to salt and water retention. The natriferic steroid aldosterone promotes salt and water retention by activation of the ATP regulated potassium channel (KArp) via a membrane-bound, protein kinase C dependant mechanism and a slower genomic pathway. We investigated whether oestrogens have similar effects. Normal human colonic epithelium was mouhted in' Ussing chambers and the transmembrane potential was clamped to OmV by application of a short circuit current (S.C.C.) which is then a measure of electrogenic ion transport. The apical membrane was rendered electrically silent by treatment of the mucosal surface with the ionophore nystatin and a mucosa to serosal potassium gradient was established. Under these conditions the S.C.C. is a measure of potassium ion transport across the basolateral KAa. P channel. Subequent addition of the oestrogen B-oestradiol produced a 7.2 + 0.8% increase in S.C.C. (n=5) within 30 min. 17-a-oestradiol was more effective, it produced a 10.2 + 1.2% increase within 30 min (n=4). Addition of chloride ions to the apical membrane of the nystatin treated colon causes cell swelling, this activates a calcium regulated potassium channel (Kc,) and inhibits the KAr r channel. Under these conditions addition of 17-a-oestradiol caused a 7.1 + 1% decrease in S.C.C. after 30 min (n=4). These effects on the KA~ v and the Kca channels were abolished by treating the basolateral membrane with the diuretic amiloride (P< 0.01) or with sodium free Krebs (P<0.01), this suggests that these effects occur due to alkanization of the cell by stimulation of the Na+/H § exchanger. Upregulation of the KAT P channel promotes salt retention by generating a favourable electrochemical gradient for sodium absorbtion by surface cells. Inhibition of the Ken channels prevents chloride secretion by colonic crypt cells. Chloride secretion causes loss of salt and water from the colon. Oestradiol simultaneously activates sodium absorbtion and inhibits chloride secretion thus causing net salt and water retention in human colon. This provides a model for oestrogen effects on salt absorbing epithelia and has important implications for the management of fluid balance disturbances associated with high oestrogen states. The aim of the study was to estimate the incidence of symptomatic ulcerative colitis (UC) and Crohn's disease (CD) prospectively from 1st October 1991 to 30th September 1992 in the Eastern Health Board region. Cases were defined using the Lennard-Jones criteria and were identified primarily by systematic examination of hospital medical records. Risk factor data was obtained via telephone interviews with the incident cases. Data on each patient were collected using a standardised questionnaire by one researcher. A total of 135 UC cases and 56 CD cases were identified during the 12 month study period. Using population figures from the 1991 census, crude incidence rates for the diseases were 11.03/100,000 in UC and 4.58/100,000 in CD. Mean age at diagnosis was 41.2 yr in UC (range 16-85 yr) and 35.6 yr in CD (range 10-76 yr). Males accounted for 58.2% of UC and 39.3% of CD cases. The presence of blood or mucous in stools was the most frequent presenting symptom in all cases: 86.6% in UC and 50.0% in CD. Risk factor data demonstrated that 26.1% of UC and 51.8% of CD cases were smokers (males and females) and that 48.3% of UC and 50% of CD cases had ever used the oral contraceptive pill (females only). A positive family history of inflammatory bowel disease (in first degree relatives) was found in 1.5% of UC and 16.1% of CD cases. Conclusions: This is the first study to estimate the incidence of UC and CD in the Irish Republic, from a study population of over 1 million people. Demographic and risk factor data among Irish cases are consistent with previously published studies. These incidence rates may be used as a baseline to detect any change in the risk of inflammatory bowel disease in future studies. Apoptosis (a form of programmed cell death) of lymphocytes is an important mode of immune regulation in the mucosal immune system. Apopt0sis also has a critical role in pathologic states including tumour-derived immune suppression, inflammation and acquired immune deficiency syndrome. The only physiological, receptor-mediated mechanism for lymphocyte apoptosis that has been characterized is the Apo-l/ FAS (FAS) receptor-ligand pathway. We have used a partially purified oesophageal tumour-derived factor to demonstrate the presence of a hitherto unrecognised, non-FAS mechanism of apoptosis in mucosal and systemic lymphocytes. The factor is selective, non-toxic, and is inhibitory to activated, proliferative but not resting lymphocytes. The FAS antigen-deficient MRLllpr mice and their normal congenic control strain were obtained and bred under I.J.M.S. July, August, September, 1995 conventional conditions. The MRLIIpr mouse has a specific genetic mutation leading to defective apoptosis through the FAS pathway and develops lymphoproliferative and autoimmune disorders. A partially purified preparation of oesophageal tumour-derived inimune suppressor factor that has been shown to be free of all known cytokines was tested in dose-ranging studies on cell proliferation and apoptosis using lymphocytes from the mutant and control mice. Proliferation was shown using thymidine incorporation and apoptosis demonstrated by flow cytometry (propidium iodide/Hoechst reagent). The lpr mice were confirmed to have deficient FAS signalling as shown by defective labeling with FITC-tagged monoclonal anti-FAS and by defective inhibition of proliferation in functional assays with FAS antibody. Lymphocytes from the congenic control strain expressed the FAS receptor appropriately and had no functional deficit. When the responsiveness of lymphocytes from the two strains of mice to the tumour-derived immunesuppressor were compared, it was found that suppression leading to apoptosis occurred equally in a concentrationdependent manner in both the FAS-mutant (lpr) and the FASpositive congenic control. Conclusion: There exists a non-FAS-mediated pathway for triggering apoptosis in lymphocytes as shown by the ability of a human tumour-derived immune suppressor to mediate apoptosis equally in FAS-negative (mutant) and FAS-positive murine lymphocytes. This system can be now used to probe the non-FAS pathway and associated secondary signalling events. The malignancy of colorectal cancer (CRC) is related to its ability to disseminate as evidenced by the observation that the majority of CRC deaths are due to the development of metastases. Urokinase-type plasminogen activator (u-PA) is a serine protease implicated in tumour invasion. We previously employed semi-quantitative immunohistochemistry to demonstrate that u-PA antigen level is of prognostic importance in Duke's B colorectal cancer (CRC) 1. The aim was to quantitate u-PA levels in colorectal tissue and to determine the relationship between u-PA level, clinical features, tumour stage, tumour grade and patient survival following CRC resection. There were 105 CRC cases (63 males, 42 females mean age 68 yr, range 36-83). Tissue level of u-PA antigen was measured on detergent extracts of CRCs and corresponding colorectal mucosa remote from the cancer using an EMSA assay (American Diagnostica). There was a wide variation in the range of u-PA levels in normal mucosa and a significant difference between normal colonicand normal rectal levels (p--0.001). Results are expressed as ratio of cancer u-PA to normal mucosal u-PA to correct for this variation, u-PA ratio was independent of age (p=0.6), gender (p=0.23) tumour site (p=0.77), tumour size (p=0.75) or tumour stage (p=0.16). u-PA cancer/normal ratios were greater in poorly differentiated tumours (p=0.03). Patients with a high cancer/normal u-PA ratio had a significantly shorter survival than those with lower ratio (logrank test p=0.0009). No. 3 Cox regression analysis identified tumour stage (relative risk [RR] 2.6, p<0.0001) and cancer/normal u-PA ratio (RR 2.3, p=0.05) as independent prognostic features. Conclusion: u-PA antigen measurement is an independent variable which is predictive of survival in colorectal cancer. In recent years, the efficacy of treatments aimed at the inexpensive, one week Bologna regime (omeprazole 20 mg daily + clarithromycin 250 mg b.i.d. + metronidazole 400 mg b.i.d.) and the two week Bordeaux regime (omeprazole 20 mg b.i.d. + amoxicillin 1 g~am b.i.d + clarithromyein 500 mg b.i.d.) achieve eradication in 90-100% of cases. There is as yet, however, no standardised approach to eradication therapy. A first line treatment should be reliable, well-tolerated, inexpensive and efficacious. The aim was 1) to assess the suitability of the Bologna regime as a first line eradication treatment. 2) To determine the factors that lead to treatment failure. 3) To evaluate the Bordeaux regime as a second line eradication treatment. Subjects with H.pylori-associated duodenal ulceration (DU) or non-ulcer dyspepsia (NUD) were recruited at endoscopy. H.pylori status was assessed before and 4 weeks after treatment by histology (antral + corpus x 2), culture (antral + corpus) and CLO-test (antral); subjects were positive if 2 or more tests were +ve and negative if all tests were -ve. All subjects were treated with the Bologna regime. 162 subjects were enrolled (79 male), 141 NUD and 21 DU, mean age 49 yr (range 18-78). 150 patients completed the followup. H.pylori was eradicated in 121/150 (80.6%). Pre-treatment sensitivities were available in 20 of the 29 patients in whom treatment failed. 18/20 (90%) had primary metronidazole resistance, 1/20 had metronidazole and clarithromycin resistance and the remaining patient was sensitive to both antibiotics. 14 of the 29 subjects were subsequently treated with the Bordeaux regime. H. pylori was eradicated in 13/14 (92.9%). Conclusion. The inexpensive, Bologna regime eradicated H. pylori in 80.6% of patients. Primary metronidazole resistance is an important factor in treatment failure. The more expensive Bordeaux regime is a highly effective second-line treatment. A prospective comparison of laparoscopy and combined imaging (computed tomography and ultrasound) was made in the pre-operative staging of distal oesophageal and gastric cancer in patients who were selected for surgery. Patients with clinically overt metastases or a contraindication to major surgery were excluded from the study. All patients underwent chest radiography, computed tomography of mediastinum and abdomen, ultrasonography of abdomen and laparoscopy. Of 145 patients assessed consecutively, 39 (26.89%) had metastatic disease to the liver or peritoneal cavity, 35 of whom were detected pre-operatively. Metastases were detected by laparoscopy in 31 patients, and by combined imaging in 16 (ultrasound 11, contrast enhanced CT Scan 14). The distribution of metastases detected pre-operatively were peritoneal cavity in 23 (laparoscopy 23 versus combined imaging 7) and hepatic 12 (laparoscopy 9, combined imaging 7). Laparoscopy was significantly more sensitive than combined imaging in detecting metastases in patients with adenocarcinoma (lapar0scopy 29, combined imaging 12) than in those with squamous cell carcinoma (laparoscopy 2, combined imaging 4). Conclusion. Patients with cancer of the oesophago-gastric region who are selected for surgery, have metastases to the peritoneal cavity more frequently than to the liver. These metastases are more frequently detected by laparoscopy than by imaging methods. The addition of laparoscopy to the staging protocol prevented unbeneficial thoraco-abdominal exploration in 28 patients. Laparoscopy should be used in the assessment of these patients immediately prior to performing excisional surgery. Laparoscopic surgery has appeal for the performance of intraabdominal procedures with minimal invasion, however the range of its applicability requires further definition. The utilisation of laparoscopy in colorectal surgery has been limited by concern regarding the adequacy of 'clearance' of malignant disease with this technique. A series of patients in whom laparoscopic mobilisation of the colon was performed to assist colectomy, facilitating a less invasive abdominal incision, is reported. From July 1994, 16 patients have undergone a laparoscopically-assisted colectomy (LAC). (10 males, 6 females; mean age: 57.5 yr, range: 20-81 yr). The indication for surgery and the procedure performed are listed in the August, September, 1995 With the patient in the Lloyd-Davis position, pneumoperitoneum was induced in the standard fashion. Laparoscopic access was via 4 ports (10ram x 3; 5mm x 1). Depending on the procedure, the splenic flexure alone, or with the transverse colon and hepatic flexure, was mobilised laparoscopically. Thereafter, access was via a transverse lower abdominal incision for completion of the procedure. In all patients, colonic mobilisation was successfully achieved at laparoscopy. There were no complications related to this procedure. In 1 patient, a major pelvic bleed occurred during the open dissection. Mean operating time (total) was 222.5 min (range: 180-380). Colonic mobilisation was completed in less than 1 hr in all cases. 3 patients developed specific postoperative complications (anastomotic dehiscence: 1, Pouch sepsis / excision: 1, pelvic abscess: 1). Initial results indicate reduced post-operative respiratory complications, analgesia requirements and hospital stay in patients undergoing laparoscopically assisted colectomy. Colonic fluid hypersecretion is an important and life threatening consequence of diseases such as inflammatory bowel disease and infectious diarrhoeas.-Secretion of chloride ion occurs in the crypts while sodium and water follow passively. The chloride ions leave the cell through an ion channel, this involves a net loss of positive ion from the apical membrane which must be balanced by an exit of positive across the basolateral membrane. In this studywe investigated the role of basolateral potassium channnels in regulating charge balance thus maintaining secretion of chloride and ultimately salt and water. Samples of colon were obtained from humans, rabbits and mice. Sheets of isolated epithelium were mounted in Ussing chambers. The short circuit current was measured under voltage clamped conditions. Bathing solutions were adapted to study chloride secretion and potassium recycling in isolation. Addition of forskolin (a secretagogue which increases intracellular cAMP) produced an immediate and sustained increase in chloride ion secretion in human (A S.C.C. = 45 + 8 uA/cm 2 n=10) rabbit (A S.C.C. = 30 + 5uAJcm 2 n=4) and mouse (A S.C.C. = 30 • 5uA/cm ? n--4). Treatment of the basolateral membrane with 10 umol tetra-pentyl-ammonium (T.P.A. an inhibitor of the Kca channel) produced a complete inhibition of the response in all three species, wheras tolbutamide (an inhibitor of the KAr p channel) produced less than 5% inhibition in all colons. Addition of carbachol (a secretagogue which increases intracellular calcium) produced an immediate but transient increase in chloride secretion which was completely inhihited by pre-treatment with T.P.A. Chloride secretion in human and other mammalian colons is dependent upon potassium exit through the Kca channel on thc basolateral membrane, this is essential to maintain charge balance within thc cell. Fluid secretion into the lumen of the colon may be prevented by inhibition of these potassium channels. The basolateral Kc~ channel may be a useful target for the pharmacological control of fluid secretion in the colon. Anal canal pressures may be raised in patients with acute anal fissure. A prospective study of the effect of parturition on the pelvic floor has allowed study of the hypothesis that anal sphincter hypertonia predisposes to postpartum anal fissure. 313 primigravid women were studied. Anal manometry was performed six weeks before and after delivery in 209 women. Postpartum studies only were performed on a further 104 women. A bowel symptom questionnaire and detailed obstetrical record were kept. Anal fissure was diagnosed on history and examination. 29 (9.3%) women developed postpartum anal fissure. Antenatal anal canal pressures were identical in those who did and did not develop fissure. Postpartum resting and squeeze anal pressures were decreased equally (table). Postnatal constipation was more common in those with fissure (62%) than those without (29%, chi 2 5.2, p<0.02). Caesarean section (11%), operative delivery (23%), perineal tear (22%) or epidural anaesthetic (54%) did not effect the incidence of fissure. Conclusion. Anal canal pressures are normal prior to development of postpartum anal fissure, and decrease postpartum. Gastrointestinal (GI) infection is a common cause of morbidity in young animals. Evidence is emerging that passive oral immunisation of the G.I. tract with specific immunoglobulin (Ig) provides protection against intestinal pathogens. Hens immunised against such organisms lay eggs containing large amounts of specific Ig. The aim of this study was to test the ability of specific hen egg Ig to protect against Salmonella infection, in the young calf. Twelve Isa Brown hens were immunised with heat inactivated Salmonella. An ELISA was developed to measure specific Ig levels, which reached a peak 20 days post immunisation. Specific Ig rich eggs were collected, the yolks harvested and pooled. An animal model of Salmonella induced gastroenteritis was developed using young calves. Forty two animals were divided into three groups. Two of these groups acted as controls, one receiving no egg yolk supplement, while the other received normal egg yolk from non-immunised hens as a supplement. The third group received a supplement of specific-Ig rich egg yolk from immunised hens. After five days each animal was orally challenged with 5 x 108 live Salmonella. In the 2 groups of control animals, 14/16 animals given no egg supplement, and No. 3 9/13 animals fed normal egg, succumbed to infection within three days of challenge. However, in the group fed Ig rich eggs, only 4/13 animals succumbed to infection after challenge. These preliminary results suggest that specific hen egg yolk Ig has a potential use in passive oral immunisation against GI infections. An inverse relationship between appendicectomy and ulcerative colitis (U.C.) and smoking and U.C. has been proposed. Our study examines the frequency of common surgical interventions and of smoking in a group of IBD patients attending a twice weekly G.I. clinic. 440 patients were interviewed in a prospective, questionnaire based case control study. The subjects comprised 143 patients with U.C., 117 patients with Crohn's and 175 controls derived from aia orthopaedic traumatology clinic matched for age, sex and socio-economic group. Subjects were questioned on all previous surgery and on smoking history. The appendicectomy rate amongst controls was 18.2% (32/ 175) which was significantly greater than that of U.C. patients 10.1% (15/148p < 0.05) 11 of the U.C. patients had appendicectomy prior to disease onset. The appendicectomies were evenly distributed through the 3 categories of disease extent (proctitis, left sided and extensive colitis). There was no significant difference in appendicectomy rate between Crohn's patients and controls. The 3 groups had comparable rates of tonsillectomy and cholecystectomy. 83.2% of the U.C. patients were non-smokers at the time of diagnosis in contrast to 50.4% amongst the Crohn's patients (p <0.01). Conclusions: U.C. is associated with significantly lower appendicectomy rate than in controls. This is not true for other surgical procedures unrelated to the treatment of U.C. The role of the appendix in the evolution of U.C. requires definition and clarification with large multicentre studies. The immune response to gliadin has been proposed to play a central role in th e pathogenesis of coeliac disease. In this study, sensitivity to gliadin was investigated in 20 coeliac and 14 normal individuals. PBMCs were cultured with soluble gliadin and latex bead coated gliadin (latex-gliadin) at varying concentrations. PPD was included as a control antigen. Culture supernatants were removed after three days and cytokines (IL4,IL5,IL6.IFN)') measured using capture ELISAs. Proliferation was assessed on day 6 by measuring thymidine incorporation. Gliadin induced proliferation in cell cultures from 16 of twenty coeliac and 9 of fourteen normal individuals. IL6 production was seen in 11 of thirteen coeliacs and 8 of nine normal controls and these responses were independent of proliferation. Gliadin failed to induce IL4 or IL5 production. Whereas PPD induced IFN'r production in association with proliferation in all individuals, gliadin induced IFN), production was less frequently observed. Of 15 CD patients, four (all with active disease) demonstrated gliadin induced IFN T production (with both soluble and latex-gliadin). In the normal controls an IFN~ response was only observed with latex-gliadin in two of 8 individuals tested. Latex-gliadin also seemed more potent at inducing proliferation with 4 subjects (3 normals, 1 coeliac) responding to this preparation alone. In preliminary experiments, cells cultured with gliadin for ten days when restimulated with gliadin demonstrated a suppressed response: in this cell proliferation, IL4 and IL6 production was inhibited but IFN~/ was increased. These results indicate that gliadin specific T cells are present in the peripheral blood of both coeliac and normal individuals. It is postulated that cells may be of the Th2 phenotype in normal subjects and coeliac patients with inactive disease (due to the observed increase in IL6 in the absence of IFN~t production) whereas in patients with active disease Thl responses may predominate. ADULT COELIAC DISEASE IN AN IRISH PROVINCIAL HOSPITAL B. Twohig, K. Gaynor, R E O'Regan. St. Joseph's Hospital, Clonmel. Previous reports on adult coeliac disease in Ireland came from teaching hospitals. These often serve as tertiary referral centres with a possible resulting different patient type from hospitals serving a district general type function. We report on all patients diagnosed over a 10 year period in a county hospital serving a mixed urban/rural population of about 85,000. A retrospective study of all patients newly diagnosed with coeliac disease between 1/1/85 and 31/12/94 who'were over the age of 16 years was carried out. The sex, age, delay from onset of symptoms to diagnosis and mode of presentation were established and lab. parameters included haematology screen, serum calcium, serum albumin and specific antibodies were also recorded. There were 46 patients diagnosed over the 10 years at a mean age of 51 (range 16 -79 yr). The duration of symptoms prior to diagnosis ranged from three weeks to 30 yr (average 17 months). Those with mild non specific symptoms and in particular females, were most likely to have long delayed diagnosis. The principal modes of presentation were diarrhoea 26 (56%), weight loss 22 (47%), anaemia 18 (39%) of which 30% was iron deficiency, hypocalcaemia 5 (11%), steatorrhoea 3 (7%). In addition to haematological abnormalities, other abnormal lab. tests included low serum calcium in 9 of 36 tested, low serum albumin in 4 of 20 tested. Alpha gliaden antibodies were raised in 16 and normal in 10 patients with definite coeliac disease before starting on a gluten free diet. Endomysial antibodies in our population also seemed to be a poor marker for the condition, being raised in only 1 of 4 patients with definite coeliac disease before starting on a gluten free diet. Conclusions: In adults the symptoms of coeliac disease may be non specific but certain symptoms such as diarrhoea with or without weight loss, inadequately explained anaemia (particularly iron deficiency) and hypocalcaemia should suggest early small bowel biopsy to rule out coeliac disease. Our study demonstrates a very long delay in diagnosis in some patients and particularly in women of child bearing years with anaemia. _oc.e.j _j_astroen_erO_Ooj July, August, September, 1995 Despite early optimism alpha gliaden antibodies have poor specificity and sensitivity in adult patients with coeliac disease in our area and it appears that endomysial antibody may not be any great improvement. Activated MOs produce 0 2-and TNF, known tumouricidal mediators. TIMs have been reported to demonstrate impaired production of these mediators suggesting that these effector cells are supressed. We postulated that TIM cytotoxicity is not dependent on the release of these cytotoxic agents but is mediated via the release of the enzyme-arginase. Currie et al have demonstrated that arginase is cytotoxic to tumour cells by depletion of the essential amino acid L-arginine, therefore the aim of this study was to determine the role of this enzyme in colorectal tumour-derived MOs. Human peripheral blood monocytes (PBM) were isolated from aged-matched controls (CON) and from blood pre-operatively obtained from patients undergoing surgery for colorectal cancer. Colorectal tumour specimens (5-10gm) were freshly procured and digested. The percentage of TIMS was then assesse d flow cytometrically, and both PBMs and TIMs were seeded at a concentration of 2 x 105/ well. Cytotoxicity was assessed using a standard 51Cr release assay against the SW707 colorectal cell line. TNF, 0 2-generation and cytosolic arginase were also assessed. These results indicate that both PBM and TIMs have impaired production of TNF and O2-, however, cytotoxicity is enhanced above CON and is associated ~,th a paralleled increase in arginase activity. The arginase-dependent nature of this cytotoxicity was confirmed by addition of the inhibitor L-lysine which significantly abrogated cytotoxicity in all groups. These results suggest caution in the use of the arginase substrate Larginine in the management of colorectal cancers. M~=macrophage, O2-=superoxide anion, TNF=tumour necrosis factor, TIMs=tumour infiltrating macrophages. Grosvenor Road, Belfast BTI 2 6BJ. The antitumour effects of L-arginine has been demonstrated in a number of experimental models including mammary and transplantable solid tumours. This study was to investigate the effect of dietary arginine in procarcinogen 1,2dimethylhydrazine (DMH) induced colorectal cancer model. Colorectal tumours were produced in male Wistar rats by 20 weekly subcutaneous injections of DMH at a dosage of 20 mg/ kg body weight. L-arginine was given in a 1% solution of drinking water. Animals were sacrificed 2 weeks after the last DMH injection. Animals were divided into 4 groups. Group I was given DMH injections, Group II was given DMH and 1% arginine for 22 weeks, Group III DMH and arginine for the first 10 weeks only, Group IV DMH and arginine for the last 12 weeks. Fourteen out of 24 animals developed tumours in Group III compared to 23 out of 24 in Group I (p=0.006, Yates-corrected Chi 2 test), 19 out of 24 in Group II and 19 out of 21 in Group IV. Decreased incidence concomitant with reduced tumour area and volume were found only when dietary arginine was given in the first 10 weeks of colorectal carcinogenesis i.e. during the initiation stages. Arginine supplementation during the promotion phases appeared to significantly enhance the tumour burden. Conclusion. These results suggest that dietary arginine may be used for tumour prevention only in the early stages of colorectal carcinogenesis before tumour formation but that its effects may be harmful in the later stages of tumour development. Early experience with interferon therapy in haemophiliacs chronically infected with hepatitis C suggested that they respond as well as non-haemophiliacs to interferon alpha. The aims of this study were to examine the effects of interferon therapy in hepatitis C in an Irish haemophiliac population. We prospectively studied 29 haemophiliac patients who received 32 courses of interferon alpha 2b (IFN), using HCV-RNA PCR to guide dosage and duration of.therapy as well as to determine virological response. All patients were asymptomatic, HCV RIBA and PCR positive, and had abnormal liver enzymes. Seven had concomitant asymptomatic HIV infection; none had hepatitis B. Genotyping in 20 patients revealed Type 1 in 14, Type 2 in 2 and Type 3 in 4. Treatment was initiated with IFN 3-5 MU s.c. alt die and increased to a max of 6 MU if LFT's remained raised or PCR +ve after 3 months. Blood was drawn monthly for liver enzymes and PCR. Responders were encouraged to continue treatment for 12 months. Three patients had sustained biochemical normalisation and negative PCR on treatment of which 1 remains in remission 1 year off treatment. Fourteen became virologically negative by PCR on treatment but relapsed within 3 months of stopping interferon. Three were retreated with interferon and again responded while on treatment. Twelve had no evidence of biochemical or virological response. We conclude that response to interferon is common in an Irish haemophiliac population. However, sustaine d virological and biochemical response rates appear