April 1995 Intestinal Disorders A305

• EFFECTS OF JEJUNAL MUCOSAL NITRIC OXIDE (NO) SYNTHESIS ON SURVIVAL DURING HEMORRHAGE-REINFUSlON SHOCK IN ANESTHETIZED RATS. David Mailman. Biology Dept., Univ. of Houston, Houston, TX 77204.

Puroose : To determine if increasing or decreasing mucosal NO altered survival time during hemorrhagic shock and possible mechanisms contributing to the central role of the intestine in multiple organ failure.

Methods: The jejunal lumen was cannulated and perfused with saline + L-arginine (LARG) or N-G-nitro L-arginine (NOLARG) at a concentration (50 gM) that produces only mucosal effects (Mailman, Br. J. Pharmacol., 1994). Rats were bled to 40 mmHg for 60 min and then the remaining blood was reinfused. Net and unidirectional H20 fluxes and mucosal absorptive site blood 'flow (ASBF) were measured. Blood pressure, NO2-/NO3- and lumenal protein were used as indicators of shock, mucosal NO synthesis, and mucosal damage, respectively.

Results. Survival time was reduced by lumenal NOLARG (P<5%) to 60 _+ 11 min and increased (P<5%) to 172 + 4 min by LARG relative to lumenal saline controls at 114 + 23 min. The volume of blood not taken back after hemorrhage was 1.8 + 0 .1%BW with lumenal LARG (P<5%), 0.8 %BW with saline and 0.3 %BW with NOLARG. Blood pressure, excluding the period in which the animal died, was not significantly different among the groups. Net H20 absorption and ASBF were significantly decreased in the NOLARG group. Lumenal protein secretion was significantly increased after reinfusion in the LARG group. There were no significant differences in lumenal NO among the groups.

Conclusions. There is a protective effect of mucosa! NO during hemorrhage. The changes in gut function were due to morbidity rather than a cause of morbidity. The mechanism of protection was not determined but a downstream effect of the gut on another organ is a possibility.

• SHORT-CHAIN FATTY ACID (SCFA) ENEMAS ARE USEFUL FOR THE TREATMENT OF CHRONIC RADIATION PROCTITIS (CRPI. J.J. Mamel, M. Chen, W. Combs, I. Tanabe, Dept. of Medicine, University of South Florida, Tampa, FL.

Radiation injury to the rectosigmoid area occurs as a result of ionizing radiation therapy to malignancies in adjacent pelvic organs. The acute phase often begins during the second week of therapy due to direct crypt injury. Chronic injury occurs after a delay of 4-24 rues or more in 2-24% of cases. It is due to a progressive endarteritis which can result in long term morbidity. Medical therapy for CRP is directed to control tenesmus and rectal bleeding. The latter may be severe enough to require blood transfusions. Stool softeners, steroid enemas, 5-ASA enemas and SAS telangiectasias using laser or electrical cautery has been utilized but it cannot be effectively applied to diffuse areas of friability. SCFA enemas provide a novel approach to nurture the injured rectal mucosa since they are utilized directly from the gut lumen, not requiring vascular translocation to reach the mucosa. SCFA have been useful in treating diversion colitis and idiopathic ulcerative proctitis.

Six patient s (4M/2F) ranging in age from 61-80 yrs. received 3600- 6000 Gy external or combination internal/external radiation for carcinoma of the prostate (4), anal melanoma (1), and vaginal carcinoma (1) for T3-4NoMo lesions. All patients have received prior combinations of 5-ASA and steroid enemas, stool softeners or mucosal cautery without improvement. 2/6 patients had persistent bleeding prior to SCFA therapy requiring blood transfusions. All patients were free from measurable tumor at the time of SCFA therapy 4-15 mos after irradiation. SCFA enemas were compounded according to Harig using acetic acid 60mM, propionic acid 30mM, n- butyric acid 40mM, NaCI 22mM, titrated to a pH of 7.0 using NaOH pellets. 60 mL enemas were instilled by the patients bid for one month. Flexible sigmoidoscopy or colonoscopy was performed initially as indicated and a repeat flexible sigmoidoscopy to 60cm after 1 month. Initial endoscopic appearance was characterized by rectal telangiectasias, loss of vascular pattern, and friability with spontaneous bleeding. All 6 patients experienced clinical improvement in decreased stool frequency, tenesmus and bleeding. Two patients requiring blood transfusions prior to SCFA enemas required no further transfusions and maintained a stable hemoglobin level. Endoscopy at 1 me. showed less friable mucosa and no spontaneous bleeding.

SCFA enemas appear to be a promising form of therapy for reducing the morbidity of chronic radiation proctitis.

• EXPRESSION OF TYPE 1I CYCLIC GMP DEPENDENT PROTEIN KINASE IN RAT INTESTINE. T. Markert, U. Walter, T. Jarehau, *A: B. Vandraager, *H. De Jonge, S. M. Lohmann. Klinische Biochemie and Pathobioehemie, Medizinisehe UniversiUitsklinik Wgrzburg, Germany and * Dept. o f Biochemistry, Erasmus University, Rotterdam, The Netherlands Background: Heat stable enterotoxin (STa) and gumxylin invre,~e cGMP by activating a membmae bound gtmaylyl cydase, The intracellul& effeetor for cGMP has not yet been clm-ified. Recently we deacribed a type II cGMP dependent protein kin~e (cGK ID which was found highly expressed in intestinal mucouL Methods: cGK II mRNA expression was assayed by Northern blots ofintastinal scrapes and by in situ bybridization of paraformaldehyde perfusion fixed tissue. Recombinant cGK II was expressed in E. coil and used for imm,miTmlon of rabbits, cGK II protein expression was studied by Western Blot and immunohistochemistry. Intact brush borders were released from jejunal-ileal segments by vibration; brush border membmae vesicles were prepared by a ~ee~e- thaw procedure and differential MgS04 precipitation. Short circuit current was measured in an Ussing chamber. Results: cGK II mRNA and protein were detected in the intestinal segments fi'om duodenum to the proximal colon. Neither the stomach nor the distal parts of the colon expressed detectable amounts of cOK H. Within the intestinal segments, cOK II was found only in the epithelial cell lining, cGK H mRNA and protein d e e r ~ along the villus to crypt axis in the small intestine, whereas in cecum and proximal colon high amounts were found in the crypts. Subcollulm cGK II was localized in the apical brush border membrane. The cGMP dependent protein kinase type I (cGK I) showed a totally different distribution, being restricted to the smooth muscle cells of the lamina propria. No detectable amounts of cGK I wea'e found in the intestinal epithelium. Short circuit current, a measure of chloride tzansport, was activated by 8-Br-cGMP and STa in the ~m~l! intestine and proximal colon, but not in the distal colon where cGK H was not detected. In human and mouse intestine, the cyclic nucleotide-regulated CI- conductance can be exclusively accounted for by the cystic fibrosis t~ansmembrane conducnmce regulator (CFTR). Conclusion: These data support an important role o f cGK lI in the mediation of cGMP effects on ion transport in the intestine.

• The A n g i o g r a p h i c Study of Le f t Gas t r i c Ve in for Esophagea l V a r i c e a l Cases w i thou t R e c u r r e n c e a f te r Endoscop i c V a r i c e a l L iga t i on .

A.Matsumeto,H.Miyoshi,K.Yoshimura,K.Sugi,S.Nakajima,S.Asada,

I.Hirata,and K.Katsu

The Second Department of Internal Medicine,Osaka Medical College,

Taka tsuki ,OsakaJapan

Recently,endoscopic variceal ligatien (EVL) has been prevalent for

t reatments of patients with esophageal varices. However,the early

recurrence of varices after EVL has been in problem.The present s tudy

was car r ied out in the esophageal variceal pat ients to settle the

indication for an EVL t reatment by evaluation of angiographic

findings jus t after the treatment. M e t h o d s : 32 pat ients with

esophageal varices has been t reated with EVL.There were 18 patients

whose varices were disappeared within two times of EVL therapies.

From 1 5 of these 18 patients,the venous phase of celiac,superior

mesenterie ,and left gastr ic a r te r iogram were obtained.Then the flow

direction types of left gastr ic vein were investigated. R e s u l t : Among

these patients,11 patients have not been recognized their variceal

recurrence over 18 months.In 10 of non-recurrent patients had types

hepatopetal ,and one case had hepatofugal with a paraesophageal vein

as a shunt to the vena cava superior .On the other hand,in the

recurrence 4 patients,the flow dereetion types were all bidirectional.

C o n c l u s i o n s : The above facts indicates that if the flow direction

type of left gastr ic vein is hepatopetal jus t after the EVL treatments,

these patients have not been recognized their varieeal recurrence for a

long time.