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Background Treating perianal Crohn’s disease (pCD) is challenging with at least 60% of the patients not responding or relapsing after a year of 1st line biological therapy. Effectiveness data on 2nd and 3rd line in pCD is still quite limited. The aim of this study was to describe the effectiveness of 2nd and 3rd line biological therapy in pCD. Methods A retrospective cohort study of patients with pCD from a large tertiary center registry. We included all patients with pCD that were treated with >1 biologic while anti-TNF was the first one used. Primary outcome was defined as clinical failure of 2nd or 3rd line treatment (as defined by the clinician) or radiological evidence for disease progression by MRI or transrectal ultrasonography (TRUS). Secondary outcomes were relapse of perianal fistula or abscess. Results Registry included 893 patients with pCD who are being followed up routinely in a dedicated IBD clinic; 643 patients received 1 biological were excluded. Final cohort included 245 patients treated with ≥2 biologics. In 2nd line, 40/245 patients (16%) treated with infliximab (IFX), 128/245 (52%) with adalimumab (ADA), 48/245 (20%) with vedolizumab (VDZ) and 32/245 (13%) with ustekinumab (UST). 7/32 patients (22%) treated with UST failed the treatment, compared to 32/40 (80%), 101/128 (79%) , 37/48 (77%) patients treated with IFX, ADA and VDZ, respectively [Hazard Ratio(HR) 0.27 (0.1-0.69)]. Median time of 2nd line treatment with UST, anti-TNFs and VDZb was 2.5 (IQR=1-3[uk1] ), 3 (IQR=1-5) and 3 (IQR=1-4) years, respectively. Higher response rate in 2nd line therapy was associated with longer response duration to 1st line [OR (1.11-1.55), p 99/245 patients (40%) required 3rd line treatment (VDZ - 52/99 , UST - 47/99 (48%)) patients). 8/47 patients (17%) under UST therapy experienced clinical failure, compared to 41/52 (78.8%) patients treated with VDZ [HR 0.32 (0.12-0.68)]. Median duration (years) of third line treatment with both UST and VDZ was 3 years (IQR=3-1). Figure A shows Kaplan-Meier survival analysis to 2nd line and 3rd line failure. Conclusion 2nd and 3rd line biological therapies may be efficacious in pCD patients. Ustekinumab appears to be effective as a 2nd or 3rd line treatment. Longer duration of treatment is associated with a higher likelihood of response to 2nd line treatment.

Background The herbal extracts Curcumin and QingDai (QD, Indigo) were previously shown to be effective in mild-moderate and in moderate-severe ulcerative colitis (UC), respectively. We evaluated the efficacy and safety of a combination of curcumin-QingDai (CurQD) in patients with active UC. Methods This was a two parts trial. Part 1 was an open label study of 4 weeks CurQD in patients with active UC, defined by a Simple Clinical Colitis Activity Index (SCCAI) score ≥5 and a modified Mayo endoscopic sub-score≥2. Part 2 was a placebo-controlled trial conducted in two centers in Israel and Greece, that randomized active UC patients at a 2:1 ratio to either enteric-coated CurQD 3gr/day or an identical placebo for eight weeks. The co-primary outcome at week 8 was clinical response (reduction in SCCAI of ≥3 points) and an objective evidence of response (Mayo endoscopic subscore improvement of ≥1 or 50% calprotectin reduction from baseline). Responding patients continued either curcumin or placebo alone for additional 8 weeks as maintenance treatment. Expression of Cyp1A1 in rectal mucosa was assessed as a measure of aryl-hydrocarbon receptor (AhR) pathway activation. Curcumin purity, and indigo and indirubin content in CurQD were confirmed by LC-MS/MS. Results There were 59 patients enrolled in the two study parts. In efficacy analysis of part 1, 7/10 responded including 3/10 who achieved clinical remission. For part 2, 95 patients were screened and 42 were included and randomized (48.8% biologics and/or immuno-modulators experienced, 36.6% biologics-experienced). The co-primary outcome was achieved in 43% and 8% of CurQD and placebo patients, respectively (p=0.033). Clinical response was observed in 85.7% versus 30.7% (p Conclusion In this randomized controlled trial, combination CurQD was found to be effective for inducing remission in active UC patients. Induction of AhR may merit further study as a potential treatment target in active UC

Background Patency capsule (PC) is a recommended procedure to rule-out small-bowel stenosis before video capsule endoscopy (VCE).We examined future clinical outcomes among patients with failed-PC versus patients in whom PC had passed (passed-PC). Methods A Post-hoc analysis of two prospective cohort studies of adult patients with quiescent small-bowel CD that underwent PC between 2013-2020. The primary composite-outcome was the need for intestinal-surgery or endoscopic-dilation during follow-up in patients with or without failed-PC. Results A total of 191 patients were included (47-failed PC, 144-passed PC, median follow-up 34.12 months). Patients with a failed-PC had higher rates of the primary composite-outcome (21.3% vs. 1.4%, Hazard ratio [HR] 20.4, 95% confidence interval [CI] 4.4-94.1, p Conclusion Clinically-stable CD patients with failed-PC have worse long-term clinical outcomes than those without, independently of CD phenotype. Standalone PC may serve as a novel, safe and affordable prognostic examination to stratify patients with CD for future risk of poor outcomes and potentially guide pre-emptive management decisions.

Background Colonoscopy is the gold standard for evaluation of inflammation in inflammatory bowel diseases (IBD), yet entails cumbersome preparations and risks of injury. Existing non-invasive prognostic tools are limited in their diagnostic power. Moreover, transcriptomics of colonic biopsies have been inconclusive in their association with clinical features. Our aim was to assess the utility of host transcriptomics of fecal wash samples of IBD patients compared to controls. Methods In this prospective cohort study, we obtained biopsies and fecal-wash samples from IBD patients and controls undergoing lower endoscopy. We performed RNAseq of biopsies and matching fecal-washes, and associated them with endoscopic and histological inflammation status. We also performed fecal mass-spectrometry proteomics on a subset of samples. We inferred cell compositions using computational deconvolution and used classification algorithms to identify informative genes. Results We analyzed biopsies and fecal washes from 39 patients (19 IBD, 20 controls). Host fecal-transcriptome carried information that was distinct from biopsy RNAseq and fecal proteomics. Transcriptomics of fecal washes, yet not of biopsies, from patients with histological inflammation were significantly correlated to one another (p=5.3*10–12). Fecal-transcriptome was significantly more powerful in identifying histological inflammation compared to transcriptome of intestinal biopsies (150 genes with area-under-the-curve >0.9 in fecal samples versus 10 genes in biopsy RNAseq). Fecal samples were enriched in inflammatory monocytes, regulatory T cells, natural killer-cells and innate lymphoid cells. Figure 1 - Fecal-wash host transcriptome predicts histological inflammation. A) Study layout, B) Clustergram of fecal-wash host cell mRNA signatures, demonstrating that patients with histological inflammation (red) are clustered when measuring fecal wash transcriptome yet not biopsy transcriptomes. C-D) Principle Component Analysis demonstrating improved separation of inflamed patients based on fecal host transcriptomes. E, F) Expression of host genes in fecal washes has higher statistical power (Area under the Curve, AUC) in classifying histological inflammation compared to biopsies. D shows NFKBIA as an example, E shows the AUC of the 5% best classifying genes, F shows the overall AUC based on biopsies or washes. Gray areas have AUC>0.9. G) UMAP of cells obtained from scRNAseq of mouse small intestine fecal washes. Conclusion Fecal wash host transcriptome is a powerful biomarker reflecting histological inflammation. Furthermore, it opens the way to identifying important correlates and therapeutic targets that may be obscure using biopsy transctriptomics.

Background Episodes of acute ileitis or colitis have been associated with future development of inflammatory bowel diseases (IBD). Nevertheless, the incidence of IBD among patients diagnosed with acute intestinal inflammation is currently unknown. We aimed to assess the risk for future development of IBD based on clinical and radiographical features of patients presenting with abdominal symptoms to the emergency department (ED) and undergoing urgent computerized tomography (CT) of the abdomen. Methods We created an electronic data repository of all patients visiting the ED at our medical center and undergoing abdominal CT between 2011–2020. We searched the computerized database for patients diagnosed with Crohn’s disease (CD) or ulcerative colitis (UC) during a visit to the gastroenterology department within maximal follow-up time of 9 years from the index-admission to the ED. Statistical analysis including multivariate models identified the risk and possible predictors of patients to develop IBD. Results Overall, 1551 patients visited the ED and underwent abdominal imaging with abnormal findings. The radiographic parameters reported the most on abdominal CT scans were bowel wall thickening, fat infiltration, and abdominal free fluid. Twenty-three patients (1.5%) were eventually diagnosed with IBD (19 CD, 4 UC) at gastroenterological follow-up with a median of 60 days (IQR 30–120) from the time of the ED admission. Patients with a future IBD diagnosis were significantly younger (28 vs 51 years, p Conclusion IBD would be eventually diagnosed only in a minority of patients presenting with ileitis or colitis on abdominal CT. Younger age, diarrhea as a presenting symptom and colitis on imaging are predisposing factors.

Background Biologics are being used increasingly in the treatment of Inflammatory Bowel Disease. However, up to 40% of patients do not respond to biologics. Therefore, methods to predict response are imperative. We aimed to identify novel genes and pathways predictive of anti-TNF response in patients with Ulcerative Colitis (UC) undergoing electronic chromoendoscopy and probe confocal laser endomicroscopy (pCLE). We further evaluated the ex-vivo binding of fluorescent labelled biologics as markers of response Methods 26 UC patients starting anti-TNF therapy as standard of care were recruited. Pre-treatment colonoscopy, with electronic chromoendoscopy and pCLE (Cellvizio, Mauna Kea) by injecting intravenous fluorescein (2.5-5mls), was performed to assess disease activity. Targeted biopsies were taken for fluorescein isothiocyanate (FITC)-labelled infliximab staining and RNA extraction and gene expression analysis. Ex vivo labelling was evaluated by an automated analysis: after a first pre-processing step to remove biases, the labelled regions were identified using statistical multi-level thresholding, and evaluated as area and intensity. To assess response, the same endoscopic procedure was repeated at week 12–14 after anti-TNF. cDNA libraries were prepared using QIAseq UPX 3’Transcriptome reagents and sequenced. Normalised gene expressions were obtained through the CLC Genomics Workbench. Differentially expressed genes (DEGs) (FDR-corrected P-value Results At baseline increased binding of the labelled biologic was associated with a higher likelihood of response to treatment (AUROC81%, accuracy77%, PPV100%, NPV63%). 342 DEGs (75 up-regulated, 267 down-regulated) distinguished responders from non-responders, 76 fell within enriched pathways. Pathways related to inflammation, chemotaxis, TGF-beta signalling, extracellular matrix and carbohydrate metabolism were reduced and cell-cell adhesion increased in responders pre-treatment. Among the 37 genes with VIP>1, CRIP2, CXCL6,EMILIN1,GADD45B, LAMA4 and MAPKAPK2 were upregulated in non-responders pre-treatment and were good predictors of response (AUROC>0.7) in this cohort and validation cohort Conclusion A higher mucosal binding of the biologics before treatment was observed in anti-TNF responders. Responsive UC patients have a less inflamed and fibrotic state pre-treatment. Chemotactic pathways, involving CXCL6 may be novel targets to treat non-responders

Background Introduction: Intestinal ultrasound can be used to assess transmural healing and response in Inflammatory bowel disease (IBD). Bowel wall thickness (BWT) appears to be the most applicable parameter for defining inflammation. Other parameters include amplified color Doppler signal, disappearance of the normal bowel wall layers and proliferation of mesenteric fat. The implication of incorporating these parameters in defining transmural healing and response is not clear. Our aim was to examine the significance of combining these parameters with BWT for defining transmural healing and response. Methods: Methods A post-hoc analysis of data from, 2 trials was used to examine the correlation of the different IUS parameters on drug retention and trough drug levels. Results: Results The study cohort included, 94 patients (median age, 24.5) with Crohn’s disease., 44 patients were treated with adalimumab (ADA) and, 50 with infliximab (IFX). There was a significant correlation of failure of drug retention and terminal ileum (TI) BWT >, 3 mm (P=0.07), amplified doppler sign (p7.1, IFX>5) and terminal ileum BWT Conclusion Conclusions: Increased BWT was correlated with failure of drug retention. The addition of other ultrasonographic activity parameters did not significantly improve the predictive model. Therefore, TI-BWT should be used as the main target in the definition of transmural healing and response.

Background Mucosal healing is associated with improved short- and long-term outcomes in Crohn’s disease (CD). However, the likelihood of reaching mucosal healing differs between segments of the mucosal tract. The prevalence of panenteric mucosal healing is scarcely described. Capsule endoscopy is the prime modality for evaluation of the entire small bowel. Recently, a combined pan-enteric capsule (PillCam Crohn’s capsule (PCCD) that enables evaluation of mucosal inflammation in the entire digestive tract in a single procedure, has been developed. The main aim of our study was to evaluate the rate of panenteric mucosal healing in Crohn’s disease patients treated with vedolizumab (VDZ) after 52 weeks of therapy. The current interim report focuses on the rates of mucosal improvement after 14 weeks of VDZ treatment. Methods This was a prospective open- label observational study. Patients starting vedolizumab (VDZ) for Crohn’s disease underwent a patency capsule (PC) and if it was excreted within 30 hours, enrolled in the study. The patients were evaluated with PCCD at week 0, 14 and 52 of VDZ treatment. Mucosal inflammation was quantified using the Lewis score (LS) and the panenteric Pillcam score (PS). In addition, fecal calprotectin (FC) and CRP levels were measured at each visit. Patients without active mucosal inflammation on week 0 PCCD were excluded from further participation Results We enrolled 30 patients; 5/30 failed PC and were dropped out of the study.Twenty five patients underwent PCCD at week 0 and 19/25 (78%)% -at week14 (six patients were dropped out for either absence of active mucosal inflammation at week 0 (1 patient), treatment discontinuation before week 14 (2 patients) endoscopic activity – 1, withdrawal of consent (1 patient) or loss to follow-up (1 patient)). After 14 weeks of VDZ treatment, there was a significant improvement in LS (median of 900 (interquartile ration (IQR)- 1143)), vs 450 (IQR-750), p=0.013), ES (15 (IQR-18) vs 10 (IQR-11), p=0.012) and FC (996 (IQR-455) vs 742 (IQR -740), p=0.012), while CRP levels were not significantly different. Conclusion VDZ induces significant improvement in mucosal inflammation evaluated by panenteric capsule endoscopy, in addition to a significant improvement in FC levels, after 14 weeks of therapy *The study was supported by IIR grant from Takeda and Medtronic

Background Vedolizumab trough serum levels have been associated with clinical and endoscopic response in patients with inflammatory bowel disease (IBD), albeit findings have not been conclusive. A recent study demonstrated that higher trough vedolizumab levels before therapy escalation predict optimal outcome. Our aim was to identify whether vedolizumab trough levels predict outcome of consecutive biologic or immunomodulatory therapy. Methods This retrospective study included IBD patients consecutively receiving vedolizumab therapy between November 2014 and June 2021 at Sheba Medical Center. Only patients who lost response to vedolizumab therapy, and had available trough levels prior to therapy cessation were included. Clinical and endoscopic scores, as well as drug retention, were recorded at 6 and 12 months post switching therapy. Results Overall, 66 IBD patients (36 CD, 30 UC) who lost response to vedolizumab therapy were included. They were treated with vedolizumab for a median of 47 weeks (IQR 31–85). Seventy-two percent of patients (48/66) were priorly treated with anti-TNF therapy (infliximab or adalimumab). After vedolizumab cessation, 66.7% of patients were switched to biologic therapy. These included 25.8% (17/66) to ustekinumab, 22.7% (15/66) to infliximab, 7.6% (5/66) to adalimumab, 6.1% (4/66) to certolizumab and 4.5% (3/66) to golimumab. The remaining 33.3% of patients switched to tofacitinib (10.6%), 5-ASA derivates or immunomodulator monotherapy (12.1%) or to a study therapy protocol (10.6%). Median vedolizumab trough levels at therapy cessation were 32.25 μg/ml (IQR 12.8–61.4). Trough drug levels at therapy cessation were not associated with retention of biological or immunomodulatory therapy at 6 months [median 29.1 μg/ml (IQR 12.4–57.6) vs 33.8 μg/ml (IQR 27.9–58), p = 0.43] and at 12 months [median, 38.2 μg/ml (IQR 14.5–76.3) vs 30.1 μg/ml (IQR 11.5–45), p = 0.57] for patients who retained consecutive therapy versus those who did not. Conclusion Vedolizumab trough serum levels before therapy cessation due to loss of response do not predict retention of consecutive therapy.

Background Acute severe ulcerative colitis (ASUC) is considered a medical emergency and is associated with high rates of colectomy and even death. Intra-venous corticosteroids serve as first line therapy for ASUC. Patients who fail to respond will progress to rescue therapy, comprised of either medical therapy (with Cyclosporine or Infliximab) or colectomy. We aimed to detect factors predictive of first and second line therapy failure in patients hospitalized for ASUC. Methods The Sheba Medical Center database was screened for all hospitalized UC patients between, 2009–2019. All clinical, demographic and laboratory parameters were retrieved from patients’ medical files. All parameters were compared between patients either responding to or failing corticosteroids / second line therapy, respectively. All statistical analyses were performed using the MEDCALC software. Results 159 hospitalized ASUC patients were identified (117, 73.6% responded to IV corticosteroids;, 42, 26.4% failed first line therapy). 4 (9.5%) of those who failed first line therapy underwent colectomy. The only factor predisposing for corticosteroid therapy failure was pervious biological therapy. No statistically significant difference was observed in terms of MAYO score at admission, UC location, smoking, previous hospitalizations, hemoglobin levels or fever upon admission., 25 patients responded to second line biological therapy (15 Infliximab, 8 Cyclosporine, 2 Tofacitinib) while, 13 failed this therapy (6 Infliximab, 7 Cyclosporine). The only factors predicting second line therapy failure were previous biological therapy (p=0.01) and pancolitis at presentation (p=0.04). Conclusion ASUC patients previously treated with biologics and who had pancolitis at diagnosis are at higher risk of second line therapy failure and colectomy.

Background Recent studies have demonstrated the importance of involving patients afflicted by a chronic disease in their own medical therapy with respect to disease control, compliance and general quality of life. Patient activation refers to patients’ independence in daily activities, involvement in the therapeutic process and having the knowledge, skills, and confidence to manage their health and health care. This study examined the association between the degree of activation of patients with inflammatory bowel disease (IBD) and its effect on health indices Methods This prospective longitudinal study included patients with Crohn’s Disease (CD) or ulcerative colitis (UC) treated at Sheba Medical Center, Israel. The patients answered questionnaires weekly during a period of 8 months using an application installed on their personal mobile phone. The questionnaires addressed clinical and emotional disease parameters, patient’s sense of disease-control (IBD-control), quality of life (short IBD quality of life questionnaire (S-IBDQ) and patient activation measure (PAM), a 4-level model of health behavior. The lowest level indicates that the patient may not believe his/her role in self-care management is important; the second level indicates that the patient lacks confidence or knowledge to take an active role; the third level indicates that the patient is beginning to be active; and the at highest level, patients actively attempt (even with difficulty) to maintain health management behaviors over time. Demographic and medical information, as well as clinical data were retrieved from the medical records. Results Among 356 IBD patients, 208 (58%) answered all questionnaire items and were included in the study 159 (76%) had CD, and 49 (24%) UC, 116 (56%) women and 92 (44%) men). Median scores of the questionnaires were as follows: PAM -63, IQR- 52.9–70.8; SIBDQ- median 44, interquartile ratio (IQR) 40–75; IBD-control 62.4, IQR- 45.7–82. 2 IBD-control mental health component- 47, IQR- 41.1–53.3). PAM was significantly correlated with questionnaires associated with overall well-being, emotional status and quality of life (SIBDQ -r=0.34 p=0.0001, IBD-control -r=0.24, p=0.004 and PROMIS-mental health component r=0.48, p Conclusion Intensive weekly mobile-App based monitoring program was feasible and adhered to by most patients. PAM scores correlated significantly with their sense of disease-control and mental health scores. Hence, IBD patients’ involvement in medical therapy increases their self-management, sense of disease-control and emotional well-being.

Background The treatment strategy in IBD patients with previous malignancy is not well defined, and the available literature is scarce.The main aim of this study was to describe the outcome of IBD patients with previous malignancy or before exposure to IBD-related treatment. Methods The study cohort included IBD patients with at least one malignancy diagnosed before IBD or before initiation of IBD-related treatment (excluding 5-ASA/corticosteroids). All patients were followed at the department of gastroenterology in Sheba medical center between 2010 and 2021. Clinical and demographic data were extracted from the patients’ medical records. The main outcome of interest was a relapse of previous malignancy or the development of a second malignancy. Results From our IBD database comprising 3811 patients, we identified 86 patients with malignancy before IBD-related treatment: 38 diagnosed with malignancy 6 years before IBD (median time, IQR=3–12) and 48 - after (15 years (IQR7-22) IBD diagnosis but before exposure to IBD treatment. The most common malignancy preceding IBD was breast cancer found in 9/38 (24%) patients. Other malignancies before IBD diagnosis included: lymphoma 7/38 patients (18%), melanoma in 6 patients (16%), and leukemia in 4 patients (11%). After a diagnosis of IBD, 38/86 (44%) patients were treated with thiopurines, 28/86 (32%) with vedolizumab (VDZ), and 26/86 (30%) with anti-TNFs. 10/86 (9%) patients were further diagnosed with second primary malignancy. The median time from first to second malignancy was 7 years (IQR=16-6); 7/10 patients (70%) were exposed to thiopurines, 6/10 (60%) to anti-TNFs, and 6/10 (60%) to VDZ. The only clinical variable associated with second malignancy was female gender (9/40 (22.5%) vs. 1/46 (2.2%), p=0.005). 20/86 patients (23%) had a recurrence of malignancy (median- 3 years (IQR=3–9). The most common recurrent malignancies were non-melanotic skin cancer (NMSC) In 9 patients (45%) and melanoma in 3 patients (20%). Of patients with recurrence, 11 patients (55%) were exposed to thiopurines, 10 patients (50%) were exposed to anti-TNFs, and 5 patients (25%) were exposed to vedolizumab. Treatment with anti-TNFs was associated with recurrence of NMSC (6/25 (24%) vs. 2/52 (3.7%), p=0.047). Conclusion In our retrospective tertiary center cohort, recurrence of malignancies in IBD patients with previous neoplasia was uncommon. However, most recurrent malignancies were solid and hematopoietic malignancies. Female gender was associated with an increased risk of second malignancy, while anti-TNF treatment was associated with an increased risk of NMSC recurrence. Our findings underline the importance of rigorous dermatological follow-up in IBD patients with previous NMSC treated with anti-TNFs

IBEX provides the observations needed for detailed modeling and in-depth understanding of the interstellar interaction (McComas et al. in Physics of the Outer Heliosphere, Third Annual IGPP Conference, pp. 162–181, 2004; Space Sci. Rev., 2009a, this issue). From mission design to launch and acquisition, this goal drove all flight system development. This paper describes the management, design, testing and integration of IBEX’s flight system, which successfully launched from Kwajalein Atoll on October 19, 2008. The payload is supported by a simple, Sun-pointing, spin-stabilized spacecraft with no deployables. The spacecraft bus consists of the following subsystems: attitude control, command and data handling, electrical power, hydrazine propulsion, RF, thermal, and structures. A novel 3-step orbit approach was employed to put IBEX in its highly elliptical, 8-day final orbit using a Solid Rocket Motor, which provided large delta-V after IBEX separated from the Pegasus launch vehicle; an adapter cone, which interfaced between the SRM and Pegasus; Motorized Lightbands, which performed separation from the Pegasus, ejection of the adapter cone, and separation of the spent SRM from the spacecraft; a ShockRing isolation system to lower expected launch loads; and the onboard Hydrazine Propulsion System. After orbit raising, IBEX transitioned from commissioning to nominal operations and science acquisition. At every phase of development, the Systems Engineering and Mission Assurance teams supervised the design, testing and integration of all IBEX flight elements.

Competing claims about the level of ignorance, or knowledge, among the current Nexus generation are addressed. The core of the paper is a theoretical analysis of ignorance in the knowledge society. Specifically, the knowledge-ignorance paradox suggests that the intense specialization demanded by a knowledge economy militates against a broader information society and gives rise to "reading reluctance." To provide evidence for this analysis, the results of a small-scale study testing the idea of a "functional knowledge deficit" are presented. Students were asked to identify metaphorical terms that are commonly used without definition in newspaper captions. The results revealed that students could only identify about 30% of these common expressions, and that they did not do better with terms derived from computers or the popular culture. Significant differences were also found between male and female responses. Both the implications of the findings and further avenues of research are discussed.

The objective was to develop an enzyme-linked immunosorbent assay (ELISA) for the detection of Cryptosporidium parvum IgG in the serum of cats. The ELISA was an indirect ELISA using soluble C. parvum oocyst antigens and a peroxidase-labeled anti-feline IgG secondary antibody. Sera from cats with Toxocara felis, Giardia spp., Aelurostrongylus abstrusus, Isospora felis, Isospora rivolta, Toxoplasma gondii, or Taenia spp. infections were assayed in specificity studies. Following optimization, the ELISA and fecal examination for oocysts were performed on samples from 170 client-owned or humane society source cats and 1 cat inoculated orally with C. parvum oocysts. Cryptosporidium parvum oocysts were detected in feces (4/170; 2.4%), and C. parvum IgG was detected in serum (26/170; 15.3%) from naturally exposed cats. The seroprevalence data suggest that some cats in the geographical area studied were exposed to C. parvum, but persistent oocyst shedding was less common. The ELISA is not useful for predicting oocyst shedding in individual cats.

An in vitro model of Cryptosporidium parvum infection was developed utilizing an adherent human intestinal epithelial cell line HT29.74. The efficacy of potential immunologic therapy in the form of Cryptosporidium-specific hyperimmune bovine colostrum was evaluated for the ability to inhibit in vitro infection. Oocysts were purified from stool of chronically infected AIDS patients. Hyperimmune colostrum obtained from cows immunized with Cryptosporidium and nonimmune conventional colostrum were evaluated. oocysts (10(5)-10(6)) were pre-incubated with either hyperimmune colostrum, conventional colostrum, or saline as control, for 15 min at room temperature than applied to a 70% confluent monolayer of HT29.74 cells. Cryptosporidium schizonts were identified and counted per 1,000 HT29.74 cells under oil immersion after 24 h. In the presence of hyperimmune colostrum, parasite infection was inhibited by 82% (p less than 0.001), and the presence of conventional colostrum, infection was inhibited by 67% (p less than 0.001). Treatment with the soluble fraction of hyperimmune colostrum resulted in 69% inhibition (p less than 0.001) compared to the soluble fraction of conventional colostrum which resulted in only 17% inhibition (p = NS). In vitro Cryptosporidium parvum infection of the differentiated human enterocyte cell line HT29.74 is a viable method for screening immunologic therapies. Hyperimmune bovine colostrum was highly inhibitory of Cryptosporidium infection in vitro and its soluble fraction remained significantly inhibitory while the soluble fraction of conventional colostrum did not.

In vitro agar culture patterns of bone marrow cells in acute myeloid leukemia may show several growth patterns, including cultures where no colonies or clusters develop, cultures with varying numbers of clusters and no colonies, or colony and cluster formation with an extremely high ratio of clusters to colonies. Twelve cases of carcinoma of the lung are described, of which two show an in vitro growth pattern of cluster formation alone, characteristic of that seen in acute myeloid leukemia. The remaining ten patients showed slightly reduced colony numbers compared to normal.

A survey of returning and non-returning students at Scarborough College was undertaken to investigate the problem of declining retention rates at Ontario Universities. This study describes the opportunities available to and chosen by non-returning students, and then seeks to evaluate various hypotheses that have been proposed to account for enrolment attrition. Consistent with prior research, there was no evidence found to support the claim that students are being forced out of university by an inability to afford the costs of their education. Instead, the results suggested that enrolment attrition is produced by a combination of an attenuation of the student's commitment to obtain the degree and the student's awareness of and access to viable alternatives. As a result of a pervasive sense of uncertainty about the market value of a university degree, many students are willing to assay other opportunities. Specifically, those students who went to work appear to have been "pulled out" of university by the availability of a credible job. In addition, the contingencies affecting the commitment of the returning students and the students who transfer to other educational institutions are discussed. The conclusion addresses the relationship between financial circumstances and access to university, and raises questions about future enrolment patterns.

Gastric carcinoma was detected nine, 10, 18, and 21 years after the biopsy diagnosis of atrophic gastritis in four patients of a group of 40. The gastritis was presumed to be of the simple type. Tests of vitamin B(12) absorption in three patients gave normal results, no gastric autoantibodies were detected in the two patients tested, in all patients histological examination of the gastrectomy specimens revealed a multifocal gastritis differing from the diffuse gastritis of pernicious anaemia and in three patients the gastritis affected the antrum, which is unusual in pernicious anaemia. The 10% incidence of gastric carcinoma in 40 patients with simple atrophic gastritis followed for a mean period of 15 years is equivalent to that previously described in pernicious anaemia. However, in view of the relative incidence of atrophic gastritis with and without pernicious anaemia in the general adult population, it emerges that atrophic gastritis without pernicious anaemia is numerically the more important precursor of gastric carcinoma.

A prospective study on 100 homosexual male volunteers was designed to examine immunological function in relation to sexual activity and infection with the human T cell lymphotropic virus Type III (HTLV-III). Complete data were available for 71 men. In a comparison with 100 agematched heterosexual men, the study group of 100 men had a significantly higher mean serum IgG level (12.1±SD2.7 g/l vs. 10.9±2.4 g/l, p

The borohydride stabilizable binding of 2-phenylethylamine and p-tyramine to mouse brain homogenates was compared to that of tryptamine and of serotonin. The highest binding was found to be that of tryptamine, followed by that of serotonin, tyramine, and phenylethylamine. The stabilizable binding of phenylethylamine to calf midbrain (including corpus striatum) homogenates and synaptic membranes was decreased by dopamine; this amine and D-amphetamine also decreased the stabilizable binding of tyramine to rat brain homogenates. The subsynaptosomal distribution of the binding of phenylethylamine to synaptic calf midbrain fractions was also investigated. The highest binding capacity was found in the 0.8 M fraction, rich in myelin.

Incubation of p-tyramine (TRM) with rat midbrain plus corpus striatum (MB+ CS) crude synaptosomal preparations, under conditions which reduce to a minimum amine uptake, results in appreciable binding of this amine by synaptosomes. This process is inhibited by preincubation with a number of drugs active in the CNS, e.g., chlorpromazine, desipramine, d-amphetamine, and diphenhydramine. Morphine, however, does not affect this binding. The Ki for each one of these compounds, as well as the K association constant and concentration of the binding sites of TRM, were determined. These results suggest a role for TRM in synaptic transmission mechanisms occurring in the nigrostriatal system, a function which could be regulated by a number of substances representing some of the major chemical classes of centrally active drugs (CD).

We examined serum IgG fractions from 20 patients with pernicious anemia and 25 control subjects for their capacity to inhibit binding of [125I]15-leu human gastrin-17 to parietal-cell-enriched gastric mucosal cells. IgG fractions from six patients reduced gastrin binding by 45.6 +/- 12.2 per cent, as compared with a reduction of 1.8 +/- 0.7 per cent by fractions from the 25 controls. The fractions from these six patients also reduced gastrin-stimulated [14C]aminopyrine uptake by gastric cells (an index of gastric acid secretory activity in vitro) by 50.2 +/- 8.4 per cent (mean +/- S.D.), as compared with 9.2 +/- 4.1 per cent for the controls. IgG fractions from six other patients that did not reduce gastrin binding also inhibited gastrin-stimulated [14C]aminopyrine uptake, by 48.1 +/- 9.1 per cent. These reductions in gastrin binding and aminopyrine uptake were abolished by absorption of the IgG fractions with suspensions of viable gastric mucosal cells but not by absorption with liver or kidney cells. The IgG fractions did not inhibit [3H]histamine binding or histamine-stimulated [14C]aminopyrine uptake. These results suggest that serum IgG from some patients with pernicious anemia contains autoantibodies to the gastrin receptor.

In vitro agar culture patterns of bone marrow cells in acute myeloid leukemia may show several growth patterns, including cultures where no colonies or clusters develop, cultures with varying numbers of clusters and no colonies, or colony and cluster formation with an extremely high ratio of clusters to colonies. Twelve cases of carcinoma of the lung are described, of which two show an in vitro growth pattern of cluster formation alone, characteristic of that seen in acute myeloid leukemia. The remaining ten patients showed slightly reduced colony numbers compared to normal.

We examined 51 sera from patients with pernicious anaemia for their capacity to block maximal gastrin stimulation of acid secretion by isolated rodent gastric parietal cells. 14C-aminopyrine accumulation was used as the index of acid secretion in vitro. Sera from patients with pernicious anaemia gave significantly (P less than 0.005) more block of maximal gastrin stimulation of acid secretion (61.7 +/- 37.8%) than sera from 10 patients with systemic lupus erythematosus (19.6 +/- 17.7%), 10 with scleroderma (34.2 +/- 22.3%), five with rheumatoid arthritis (22.4 +/- 15.6%) or 30 from healthy persons (27.4 +/- 12.8%). Maximal histamine stimulation of acid secretion was not inhibited. The blocking factor was present in serum IgG fractions, and serum and IgG fractions gave parallel dose-response and dilution curves. The serum block was abolished by absorption with gastric mucosal cells and correlated with the presence of parietal cell surface autoantibody. We conclude that serum immunoglobulin in pernicious anaemia can block gastrin stimulation of acid secretion and suggest that this block may be mediated by competition with gastrin for surface receptors on parietal cells.

Using flow microfluorimetry (FMF), 60 sera from patients with pernicious anaemia (PA) were examined for immunoreactivity with the surface membranes of viable canine parietal cells. FMF analyses showed that the percentage of parietal cells giving a surface staining reaction with a fluorescence intensity greater than 50 arbitrary units was 44.5 +/- 17.5% for sera from 60 patients with PA compared to 13.7 +/- 2.7% for sera from 14 patients with chronic active hepatitis, 10.7 +/- 6.7% for sera from 10 patients with systemic lupus erythematosus and 16.5 +/- 4.4% for sera from 50 healthy persons. Surface staining detected by FMF was restricted to parietal cells and abolished by absorption with parietal cell enriched preparations but not by absorption with dog or rat hepatocytes, dog or rat kidney cells, human fibroblasts, human AB red blood cells or dog gastric microsomes. The intensity of the parietal cell surface staining reactions correlated with the presence of antibody reactions with parietal cell surfaces previously demonstrated by immunofluorescence microscopy but did not correlate with the presence of microsomal or intrinsic factor autoantibodies. The results provide further support for the presence of a parietal cell surface reactive autoantibody distinct from the conventional parietal cell microsomal autoantibody.

Changes in the ultraviolet (UV) absorption spectra of nicotinamide adenine dinucleotide (NAD), of its synthetic analogues, and of adenosine and its derivatives in the presence of high concentrations of orthophosphate were studied. The role of the carboxamide group of the nicotinamide moiety of NAD on these spectral changes was investigated by replacing that group with an acetyl or aldehyde group. The effect of the 6-amino group of the purine was investigated by studying the interaction of deamino-NAD and various adenosine derivatives with orthophosphate. 2,4-Dinitrophenol was also found to give a charge transfer complex with phosphate. Molar extinction coefficients (E) and association constants (K) of these charge transfer reactions were determined.

A prospective study on 100 homosexual male volunteers was designed to examine immunological function in relation to sexual activity and infection with the human T cell lymphotropic virus Type III (HTLV-III). Complete data were available for 71 men. In a comparison with 100 age-matched heterosexual men, the study group of 100 men had a significantly higher mean serum IgG level (12.1 +/- SD 2.7 g/l vs. 10.9 +/- 2.4 g/l, p less than 0.01) and a significantly lower mean number of CD4 (T4) cells (845 +/- 310 X 10(-6)/l vs. 1128 +/- 375; p less than 0.01). For the study group, seropositivity for anti-HTLV-III was present initially in 22 per cent and was associated with a higher mean level of serum IgG and lower mean number of CD4 cells. Among seropositive homosexual men a low CD4/8 ratio was attributable to low numbers of CD4 cells in those without lymphadenopathy and to high numbers of CD8 cells in those with lymphadenopathy. For the seronegative homosexual men, a low CD4/8 ratio as a result of an increased CD8 cell count was present in 12 of 60, and was associated with numerous sexual partners and semen culture positive for cytomegalovirus. In two seropositive subjects a low CD4/8 ratio due to a decrease in the CD4 cell count was predictive of the development of AIDS by some two years. For the 71 men with complete data over two years, indices of cell-mediated immunity, including mean counts of CD4 cells, the CD4/8 ratio, and score for recall of cutaneous delayed type hypersensitivity increased during the first year but not during the second year in both seropositive and seronegative subjects. These increases occurred in association with changes in sexual practices and activity, but could not be attributed to any one particular factor.

Sera from 20 of 24 patients with pernicious anaemia reacted by immunoblotting with a 65-70 kD protein in canine and rodent gastric mucosal cells enriched for 80-90% parietal cells, and in microsomal preparations derived from these cells. All 20 reactive sera were positive for parietal cell microsomal antibody demonstrated by immunofluorescence. Eighteen parietal cell microsomal antibody-positive sera from patients with unconfirmed pernicious anaemia also reacted with the same 65-70 kD protein. Serum reactivity with the same 65-70 kD protein was not seen with canine and rodent liver cells or with microsomal preparations derived from these cells. Sera from 10 patients with chronic active hepatitis, 10 with scleroderma and 10 with rheumatoid arthritis and 22 healthy persons did not react with the 65-70 kD protein. These results suggest that the 65-70 kD protein is probably the parietal cell microsomal autoantigen. A second antigen of 85-90 kD mol. wt. present only in canine gastric mucosal preparations also correlates with the presence of parietal cell microsomal antibody. However, the contribution of this second antigen to parietal cell microsomal antibody reactivity remains uncertain.

Summary: The clinical and pathological features of histiocytic medullary reticulosis (HMR) are described in five patients. Treatment by splenectomy and the MOPP routine resulted in a transient improvement in platelet and neutrophil counts and resolution of anaemia and apparent prolongation of survival in two of the five patients. The relationship of HMR to other histiocytic disorders is discussed. HMR is defined as a unique malignant histiocytosis in which malignant histiocytes engage in phagocytosis of erythrocytes, leucocytes and platelets.

Eight patients with Addisonian pernicious anaemia were given 20 mg of prednisolone daily for up to 20 weeks. Improvement in absorption of vitamin B12 as judged by the Schilling test occurred in six cases and was pronounced in four, and there was increased secretion of gastric intrinsic factor in four cases. Gastric biopsies showed regeneration of specialized gastric glands in four cases; chief cells were demonstrated histochemically and parietal cells by an immunofluorescent procedure using serum containing parietal cell antibody. There was no correlative change in titre of serum antibody to gastric parietal cells or gastric intrinsic factor. The improved absorption of vitamin B12 was not maintained after prednisolone was stopped, indicating that the regenerated gastric mucosa reverted to the atrophic state. These effects of corticosteroids on gastric function in pernicious anaemia are in keeping with their known capacity to modify damage resulting from antigen–antibody interaction.