E. coli Secreting Short Chain Variable Fragments Protect from Intestinal Salmonellosis

Yehuda Chowers1, Matityahul2, Nathan Keller3, Simon Bar-Meir1 and Martin Goldway2

Dept. Gastroenterology1, Laboratory of Microbiology3, Chaim Sheba medical Center, Tel Hashomer and MIGAL Galilee Technological Center, Rosh Pina2

Background: The indigenous bacterial milieu and secreted IgA pathogen-specific antibodies play an important role in mucosal protection.

Aims: To test the protective effect of luminal E. coli secreting Salmonella (Sal)-specific short chain variable fragments (ScFv) against intestinal salmonellosis.

Methods: Antigen-binding domain genes of an anti-Sal hybridoma were cloned into a plasmid vector and transformed into E. coli. Periplasmic fluids, bacterial and hybridoma culture media and E. coli expressing ScFv were tested for Sal binding and ability to prevent Sal invasion into HT-29 cells and IL-8 secretion. Subsequently, the effect of E. coli secreting ScFv on salmonella colonization of chick intestines was tested.

Results: Western bloting detected ScFv in bacterial periplasmic fluids and culture media. Both hybridoma culture fluid and periplasmic extracts from E. coli transformed with ScFv constructs detected unfractionated bacteria. ScFv periplasmic extracts inhibited invasion of Sal into the cells (2.6 fold) similar to hybridoma supernatants (2.7 fold). IL-8 secretion was similarly inhibited. Coculture of

4.5 X 108 ScFv expressing E. coli with 4.5 X 105 Sal resulted in a 3.2 fold decrease in Sal invasion. Subsequently, newly hatched SPF chicks were inoculated with 107 Sal and 107 E. coli transformed with either ScFv constructs or control plasmid. After 24 hrs Sal were quantitated in cecal homogenates. ScFv-expressing E. coli but not controls significantly reduced Sal CFU in the chick’s intestines.

Conclusions: Indigenous bacteria expressing ScFv may protect from intestinal pathogens. The results suggest that such approach may serve as a novel method for mucosal protection.

Allicin Inhibits Proinflammatory Cytokine Secretion from Rat Hepatic Stellate Cells

E.Sakhnini, Y. Chowers, E. Haramati, S. Bar-Meir and A. Lang

Dept. Gastroenterology, Chaim Sheba Medical Center, Sackler School of Medicine, Tel-Aviv University

Background: Activated hepatic stellate cells (HSCs) have been implicated in hepatic inflammation, a condition that often precedes and coexists with fibrogenesis, through their ability to secrete cytokines and chemokines as well as express adhesion molecules. Garlic is widely used in complementary and alternative medicine for a varaity of diseases including liver diseases. Allicin, an active component of garlic has anti inflammatory properties.

Aim: To assess the effect of allicin on inflammatory response of HSCs.

Methods: Rat HSC cell line and rat primary HSCs were used. Cells were incubated with allicin. Unstimulated, TNFa stimulated and IL-1 b stimulated cells were assessed for the secretion of IL-1 b, TNFa and IL-8 using ELISA.

Results: Allicin markedly inhibited the secretion of IL-1 b, TNFa and IL-8 from HSC. This effect resulted in a 6-fold suppression of IL-1 b from baseline, 12 fold supression of TNFa and 20 – fold suppression of IL-8 from baseline. Suppression effects of allicin were apparent following TNFa and IL-1 b stimulation as well. HSCs cell line results were confirmed using primary HSCs.

Conclusion: Allicin has a potent immunomodulatory effect on HSCs resulting in suppression of proinflammatory cytokine secretion.

DELAYED INSERTION OF PEG: A PROSPECTIVE, CONTROLLED STUDY OF NEW APPROACH.

Galia Abuksis, Meli Mor, Shlomit Plaut, Gerald Fraser, Yaron Niv

Department of Gastroenterology, Rabin Medical Center, Beilinson Campus, Tel-Aviv University, Israel.

Background PEG is the technique of choice for long-term enteral nutrition. Though described as safe and simple, PEG has been associated with significant morbidity and mortality.

Aim. We prospectively compared the outcome of 2 strategies, in 2 different periods: PEG insertion during hospitalization (upon request), and our new approach of PEG insertion 30 days after hospital discharge.

Methods. We compared 3 groups of hospitalized patients in a prospective, controlled study. Group A of 61 consecutive patients, referred for PEG insertion between 1/1/97 and 31/12/98 and were scheduled for PEG closed to the physician request, while still hospitalized. Group B of 66 consecutive patients referred for PEG between 1/1/99 and 30/6/00, and were scheduled for PEG 30 days after discharged. Group C was a control group of 314 patients matched for age, sex and medical history that were hospitalized between 1/1/99-30/12/00 with diagnosis of malnutrition. We compared 30-day mortality rate in 3 phases: Thirty-day mortality rate since admission for the 3 Groups, Thirty-day mortality rate since PEG request for Groups A and B, and Thirty-day mortality rate since PEG insertion for Groups A and B.

Results. Thirty day mortality since PEG request decreased by 40% in Group B in comparison with Group A (P = 0.01); and since PEG insertion decreased by 87.5% in Group B in comparison with Group A (P = 0.000).

Conclusion. PEG insertion during hospitalization increased mortality and should be prevented. Grace period of 30 days may prevent mortality, and supply long term enteral nutrition.

Long-term use of Percutaneous Endoscopic Gastrostomies:

A survey of duration of use and level of maintenance

Rina Dolinsky, Jesse Lachter, Ron Reshef

Western Galilee Medical Center, Nahariya, Israel

Introduction: Few long-term guidelines have been established for PEG patients. Late complications include malfunction, skin breakdown and infections.

Patients and Methods: Patients were identified by visiting nearby consenting chronic care facilities. 100 patients with PEGs for at least 6 months were included. Data collected regarded demographics, indications for PEG, duration of PEGs, and brand, and scored evaluations of the PEGs as compared to photos.

Results: Ages were mean 70 +/- 21years, 62% were women. Duration of use of the current PEG ranged from 1-27 months. The indications for PEG use were dementia 42%, CVA 33%, trauma 25%. PEG tubes were judged normal in 29%, friable in 58%, semi-obstructed in 12%. Only 35% of the stopcock valves were in good condition. Fungus was found in 51%. The post-trauma victims had better condition PEGs in terms of clean skin, tube and valve, all at p<0.001 compared to the CVA and dementia patients in the other ten sites. Skin breakdown became common, as was decayed PEG tube and stopcock, all beginning about seven months after PEG insertion, (Spearman’s coefficient at p<0.01).

Discussion: The care of patients’ PEGs was sub-optimal for most non-trauma patients. Trauma patients benefited from much more expensive care. Experts who insert PEGs may be in a position to improve the care of the patients beyond the insertion and peri-operative management.

Conclusion: Long-term feeding by PEG might be improved by on-site periodic follow-up by experts, possibly nurses, and by guidelines which would mandate PEG replacement, probably at about six month intervals.

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THE EPIDEMIOLOGY OF IBS AMONG ISRAELI JEWISH ADULTS

1Ami D. Sperber, 2Pesach Shvartzman, 3Michael Friger, 1Alex Fich

1Department of Gastroenterology, Soroka Medical Center, Beer-Sheva , 2Department of Family Medicine and 3Epidemiology Unit, Ben-Gurion University of the Negev, Beer-Sheva, Israel

Objective: The prevalence of IBS depends upon the country studied and the criteria used. Using Rome I criteria estimates of IBS prevalence ranged from 8%-20%. The prevalence using Rome II is lower. In a previous pilot study with Rome I criteria we estimated the prevalence of IBS among Israeli Jewish adults at approximately 5%. The purpose of the present study was to determine the prevalence of IBS in a large, representative sample of Israeli Jewish adults, using the Rome II criteria. Methods: A questionnaire was administered by telephone to a representative sample of the adult Israeli Jewish population provided by the Israel Ministry of the Interior. Results: We attempted to reach 1,839 individuals, of whom 1,221 were successfully contacted. Of those, 221 refused to participate in the survey: response rate = 81.9%. The final study population was 981 individuals with complete, valid data. The mean age was 45.0 years and 55% were females. In all, 2.9% had IBS (females: 3.7%, males 1.8%, p=0.08). 25.6% of the respondents had a functional lower GI disorder (females: 32.1%, males: 17.7%, p<0.0001). IBS was not significantly associated with age, level of education, income, work status, family status or religiosity, but significantly more unemployed individuals had a functional lower GI disorder (p=0.01). Conclusions: Using Rome II criteria, prevalence rates for IBS among Israeli Jewish adults are low. However, a substantial percentage of the population suffers from some form of functional lower GI disorder, most commonly functional abdominal bloating.