Phase II Trial of Copper Zinc Superoxide Dismutase (CuZn SOD) in the Treatment of Crohn'S Disease

Bovine Cu Zn SOD was used during an 8 year period as an antiinflammatory drug in 26 patients with severe Crohn's disease (CDAI 300) usually after failure of corticotherapy or when this drug was discontinued because of side eKects or infection.

This was a phase II trial during which doses routes of administration and concomitant therapies were progressively modified.

We obtained 73% good short term responses (judged upon CDAI and anatomic healing) and 82% positive results on long term evolution (the criteria were : i CDAI lower than 100 in between relapses, ii complete healing or notable improvement of lesions, iii no surgery needed, iv return to work. The acceptability was excellent with the free enzyme. Since the above described experience, published in Free Radical Biology and Medicine (1989, 7 : 145-151). we used always the same treatment schedule (SOD 8 mg/day associated with Desferroxamine -500 mg subcutaneous every 2 days). The follow-up during the 87-89 period showed that 12 are in good health without any relapse, 9 experienced one or more relapses, and showed good responses upon resumption of treatment, 5 failed to respond to treatment. all part of the initial group on which SOD treatment had already failed, and among whom 3 were lost for follow-up before 1987, and two others took up another SOD treatment which also failed. 3 new patients (2 females, 1 male) were treated since then, and all 3 had positive results (one with disappearance of ileococcal mass).

The efficacy of SOD as an antiinflammatory drug in Crohn's disease needs to be confirmed by controlled trials.     

Phase II Trial of Copper Zinc Superoxide Dismutase (CuZn SOD) in the Treatment of Crohn'S Disease

Bovine Cu Zn SOD was used during an 8 year period as an antiinflammatory drug in 26 patients with severe Crohn's disease (CDAI 300) usually after failure of corticotherapy or when this drug was discontinued because of side eKects or infection.

This was a phase II trial during which doses routes of administration and concomitant therapies were progressively modified.

We obtained 73% good short term responses (judged upon CDAI and anatomic healing) and 82% positive results on long term evolution (the criteria were : i CDAI lower than 100 in between relapses, ii complete healing or notable improvement of lesions, iii no surgery needed, iv return to work. The acceptability was excellent with the free enzyme. Since the above described experience, published in Free Radical Biology and Medicine (1989, 7 : 145–151). we used always the same treatment schedule (SOD 8 mg/day associated with Desferroxamine -500 mg subcutaneous every 2 days). The follow-up during the 87–89 period showed that 12 are in good health without any relapse, 9 experienced one or more relapses, and showed good responses upon resumption of treatment, 5 failed to respond to treatment. all part of the initial group on which SOD treatment had already failed, and among whom 3 were lost for follow-up before 1987, and two others took up another SOD treatment which also failed. 3 new patients (2 females, 1 male) were treated since then, and all 3 had positive results (one with disappearance of ileococcal mass).

The efficacy of SOD as an antiinflammatory drug in Crohn's disease needs to be confirmed by controlled trials.     

Microbiological and immunological strategies for treatment of inflammatory bowel disease.

Chronic inflammatory bowel diseases such as Crohn's disease or ulcerative colitis, affect around 1 in every 1000 individuals in western countries. They probably result from an inappropriate reaction towards the commensal microflora and are currently treated with anti-inflammatory drugs or surgery. Novel strategies aim at blocking lymphocyte recruitment and activation, improved targeting of therapeutics and modification of gut microflora.     

In vitro analysis of interferon gamma (IFN-gamma) and interleukin-12 (IL-12) production and their effects in ileal Crohn's disease

Crohn's disease is an inflammatory disease of the gut in which tumour necrosis factor (TNF) and T helper 1 (Th1) cytokines (interleukin (IL)-12, interferon (IFN)-gamma) are thought to play a major role. After the successes obtained with neutralisation of TNF, interest is now growing for therapy aiming at neutralisation of Th1-associated cytokines. Since cytokines are linked in a delicate network, in vitro cultures of ileal lamina propria mononuclear cells (LPMC) were set up for evaluation of a) IFN-gamma and IL-12 production, b) effects of rhIFN-gamma and rhIL-12 and c) effects of anti-IFN-gamma and anti-IL-12 on pro-inflammatory cytokines and IL-10 production. LPMC were isolated from surgical specimens of a total of 27 Crohn's disease and 17 caecum carcinoma (control) patients. Cells were stimulated with CD40L (which triggers myeloid CD40-expressing cells) or anti-CD3 +CD80 (which triggers T cells). LPMC from involved ileal, Crohn's disease produced, in both non-stimulated and stimulated conditions, more IFN-gamma and IL-12p70 than LPMC from non-involved tissue or from control patients. rhIFN-gamma significantly enhanced TNF production in both controls and in ileal Crohn's disease patients, while rhIL-12 enhanced IFN-gamma but not TNF production. LPMC from involved tissue were more sensitive to IL-12 than control LPMC. LP-T cell-dependent activation of monocytes was then studied by co-culture of anti-CD3/CD80-stimulated LPMC with fresh monocytes, which resulted in high IL-12, IFN-gamma, TNF and IL-10 production. The data show that neutralisation of either IL-12 or IFN-gamma with mAb in these cultures also affects secretion of the reciprocal cytokine and (in the case of anti-IL-12) also that of the anti-inflammatory cytokine IL-10. However, no effect of anti-IL-12 or anti-IFN-gamma on production of TNF, a cytokine with an important pathogenic role in Crohn's disease, could be found. Therapies aiming at neutralisation of IFN-gamma or IL-12 are therefore unlikely to replace anti-TNF, but they might provide an additive or synergistic effect.     

Hyperbaric oxygenation and drug therapy in treatment of nonspecific ulcerative colitis and Crohn's disease]

Patients with nonspecific ulcerative colitis and Crohn's disease were treated with drug therapy (prednisolone, sulphasalazine, metronidazole per os and hydrocortisone per rectum) and subjected to 12 sessions of HBO. Every 10-14 months the HBO course was repeated. After 7 years of such a treatment a gradual partial recovery of large intestine mucosa and in some cases even its absolute recovery were observed. It was most difficult to get good results when treating lesion of distal colon segments. HBO produced a good effect when the disease was diagnosed at the early stage, when the intestine injury was accompanied by hepatobiliary system disease and in teenagers.     

The Crohn's disease protein, NOD2, requires RIP2 in order to induce ubiquitinylation of a novel site on NEMO

BACKGROUND: Crohn's disease is an autoimmune inflammatory disorder of the gastrointestinal tract and is characterized clinically by dysregulation of both pro-inflammatory and anti-inflammatory cytokine signaling networks. The function of the Crohn's disease protein, NOD2, highlights the biphasic nature of the pathology of Crohn's disease. NOD2 can both strongly activate and negatively attenuate NF-kB signaling. The biochemical mechanism for this dual function of NOD2 is unknown. RESULTS: We demonstrate that NOD2 activation leads to ubiquitinylation of NEMO, a key component of the NF-kB signaling complex. This ubiquitinylation is agonist dependant, and it does not regulate proteosomal destruction of NEMO. We show the NOD2-dependent ubiquitinylation of NEMO is dependent on the scaffolding protein kinase RIP2. Crohn's disease-associated polymorphisms of NOD2 show a decreased ability to bind RIP2, and this decreased ability to bind RIP2 correlates with a decreased ability to ubiquitinylate NEMO. We map the site of NEMO ubiquitinylation to a novel NEMO ubiquitinylation site (Lysine 285) and show that this ubiquityinylation occurs in vivo. Lastly, we show functionally that RIP2-induced ubiquitinylation of NEMO is at least in part responsible for RIP2-mediated NF-kB activation. CONCLUSIONS: These data suggest that this novel mode of regulation of the NF-kB signaling pathway could be a factor underlying the pathogenesis of Crohn's disease.     

Liver: the formation and actions of aspirin-triggered lipoxins

Eicosanoids play a key role in the initiation, progression and resolution of the inflammatory response. Although most current anti-inflammatory strategies are focused on the pharmacological inhibition of pro-inflammatory eicosanoids, such as prostaglandins and leukotrienes, mounting evidence indicates the existence of potent endogenous eicosanoids able to control inflammation and orchestrate its resolution. The first eicosanoids recognized as anti-inflammatory compounds generated by our own organism were the lipoxins (LXs). More recently, a new series of carbon-15 epimers of LXs, with anti-inflammatory properties similar to those of native LXs, was identified during aspirin treatment. Since their formation is specific to this venerable non-steroidal anti-inflammatory drug, the term aspirin-triggered LXs (ATLs) was coined for these compounds. This chapter deals with the biosynthesis of LXs and ATLs in the liver, the largest solid organ/gland in the body, and discusses the most relevant actions of these lipid mediators in the context of liver inflammation and injury.     

Design,synthesis, docking and anti-inflammatory evaluation of novel series of benzofuran based prodrugs

Several new benzofuran derivatives were synthesized, via appropriate synthetic route as anti-inflammatory agents. The anti-inflammatory activity of the prepared compounds was evaluated using carrageenan rat model. Among the synthesized compounds, some compounds showed comparable anti-inflammatory activity to nimesulide, the standard drug taken for anti-inflammatory studies. Docking study of the prepared compounds was performed for the study of interaction of molecules with the active site of COX-2. Preliminary biological studies and docking gave an interesting insight, into the validity of employing benzofuran analogues as good anti-inflammatory agent.     

Thalidomide induces apoptosis in human monocytes by using a cytochrome c-dependent pathway

Thalidomide has been shown to be an effective treatment in various immunologic diseases such as Crohn's disease and rheumatoid arthritis. Its major effect is thought to be mediated by the inhibition of TNF-alpha, but the exact mechanism of action is still uncertain. Recent observations could demonstrate that the induction of monocyte apoptosis is a common feature of a variety of anti-inflammatory agents. Therefore, we investigated the role of thalidomide on monocyte apoptosis. Treatment with thalidomide resulted in apoptosis of human peripheral blood monocytes in a time- and dose-dependent manner as demonstrated by annexin V staining. Monocyte apoptosis required the activation of caspases, as combined stimulation by thalidomide together with the broad caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone markedly prevented monocyte cell death. Apoptosis was triggered by a CD95/CD95 ligand, TNF-RI, and TRAIL-R1 independent pathway with an inhibition of AKT-1 kinase and consecutive mitochondrial release of cytochrome c, followed by the proteolytic activation of initiator caspase-9 and effector caspase-3. Our data suggest that thalidomide-induced monocyte apoptosis is at least partially mediated by a mitochondrial signaling pathway and might contribute to the complex immunomodulatory properties of the drug.     

n of 1 trials comparing a non-steroidal anti-inflammatory drug with paracetamol in osteoarthritis.

OBJECTIVE--To evaluate the efficacy of paracetamol and a non-steroidal anti-inflammatory drug for symptom relief in osteoarthritis. DESIGN--Double blind, randomised, controlled trials in individual patients (n of 1 trials). Three treatment cycles with two weeks'' each of paracetamol (1 g twice daily) and diclofenac (50 mg twice daily) prepared in identical gelatin capsules. SETTING--General practices in metropolitan Sydney, Australia. SUBJECTS--25 patients (median age 64 years) with pain of osteoarthritis (median duration of disease eight years) considered by their general practitioners to require regular treatment. 20 were already taking non-steroidal anti-inflammatory drugs. MAIN OUTCOME MEASURES--Diary of pain and stiffness, function, and side effects. RESULTS--15 patients completed the study, five withdrew early but had made a therapeutic decision, and five dropped out very early. Results from 20 patients were analysed. Several patterns of response evolved. Eight of the 20 patients found no clear difference, symptoms being adequately controlled by paracetamol; five indicated a clear preference for the non-steroidal anti-inflammatory drug; two showed control of symptoms after their initial two weeks of the non-steroidal anti-inflammatory drug which continued throughout subsequent treatment changes; in five the non-steroidal anti-inflammatory drug may have been better but neither agent gave satisfactory control. After three months nine of the 20 patients had adequate symptom control with paracetamol alone. CONCLUSIONS--Of 1 studies--that is, randomised trials in individual patients--are clinically useful in deciding treatment in heterogeneous conditions which require long term symptomatic relief. In osteoarthritis many patients currently receiving or being considered for non-steroidal anti-inflammatory drugs may achieve adequate control with paracetamol.     

Visualization of Mycobacterium avium in Crohn's tissue by oil-immersion microscopy

Studies seeking Mycobacterium avium subsp. paratuberculosis in Crohn's disease by PCR have generated inconsistent findings. As an alternative, microscopy offers a number of advantages, including direct visualization of organisms in tissue. Experimental infections have demonstrated that M. avium organisms can be seen by both acid-fast staining and species-specific in situ hybridization, but because they are smaller than M. tuberculosis, oil-immersion microscopy (x 1,000 magnification) is needed. We performed a blinded search for M. avium in paraffin-embedded surgical resections from Crohn's and control subjects at two centres. Specimens were coded and subjected to acid-fast staining and ribosomal RNA in situ hybridization for M. avium rRNA. Agreement between these two methods was good (42/52 patients, kappa=0.60) and similar results were observed for patients from two centers. Together, both methods provided positive results in 10 of 17 Crohn's subjects (59%, 95% CI: 36-78), contrasting with only 5 of 35 control subjects (Odds ratio for Crohn's vs. controls=8.6, p=0.002). M. avium organisms had an intracellular localization within inflammatory lesions, but were often observed as lone organisms outside of granulomas. Using two assays in two settings, presence of M. avium organisms was strongly associated with Crohn's disease.     

Endometriosis of the large intestine]

Two cases of large intestine endometriosis are presented. The disease was diagnosed during histological examination of samples taken during surgery. Clinically one case was diagnosed as Crohn's disease, while the second as cancer of the large intestine. The authors suggest, that an extent of surgery for, tumours of the large intestine should be carefully planned, specially if the tissue specimen was not examined histologically earlier.     

Synthesis and biological evaluation of some new pyridazinone derivatives

A series of pyridazinone derivatives (19-34) were synthesized with an aim to synthesize safer anti-inflammatory agents. The compounds were evaluated for their anti-inflammatory, analgesic, ulcerogenic and lipid peroxidation (LPO) actions. The percentage inhibition in edema at different time intervals indicated that compounds 20, 26, 28 and 34 exhibited good anti-inflammatory potential, comparable with that of ibuprofen (85.77%) within a range of 67.48-77.23%. The results illustrate that 5-(4-fluoro-benzyl)-3-(4-chloro-phenyl)-1,6-dihydro-6-pyridazinone (26) and 5-(4-chloro-benzyl)-3-(4-chloro-phenyl)-1,6-dihydro-6-pyridazinone (20) showed best anti-inflammatory activity. Furthermore, activity is more in case of chloro substitution as compared with methyl-substitution. The compounds synthesized were also evaluated for their ulcerogenic and LPO action and showed superior gastrointestinal safety profile along with reduction in LPO as compared with that of the ibuprofen.     

The role of heme oxygenase and carbon monoxide in inflammatory bowel disease

Inflammatory bowel disease (IBD), including ulcerative colitis (UC) and Crohn's disease, is a chronic and recurrent inflammatory disorder of the intestinal tract. Since the precise pathogenesis of IBD remains unclear, it is important to investigate the pathogenesis of IBD and to evaluate new anti-inflammatory strategies. Recent evidence suggests that heme oxygenase-1 (HO-1) plays a critical protective role during the development of intestinal inflammation. In fact, it has been demonstrated that the activation of HO-1 may act as an endogenous defensive mechanism to reduce inflammation and tissue injury in various animal intestinal injury models induced by ischemia-reperfusion, indomethacin, lipopolysaccharide-associated sepsis, trinitrobenzene sulfonic acid or dextran sulfate sodium. In addition, carbon monoxide (CO) derived from HO-1 has been shown to be involved in the regulation of intestinal inflammation. Furthermore, administration of a low concentration of exogenous CO has a protective effect against intestinal inflammation. These data suggest that HO-1 and CO may be novel therapeutic molecules for patients with gastrointestinal inflammatory diseases. In this review, we present what is currently known regarding the role of HO-1 and CO in intestinal inflammation.     

Discoid eczema of the hands

In general it is probable that round or oval, rather defined groups of fine vesicles (or crusting that results from former vesicular eruption) recurring in the same location like a "fixed drug eruption" is a particular type of eruption deserving a distinct name. The morphological features and the clinical course of repeated recurrences, always in round or oval areas, seem to make the term "discoid eczema of the hands" an appropriate one for the disease.A feature that distinguishes discoid eczema of the hands from infectious dermatologic conditions which it most closely resembles clinically, is the response of the disease to dietary therapy. A simple elimination diet was used with good effect in 20 cases of the disease reported upon herein.     

Role of oxidative stress and antioxidant enzymes in Crohn's disease

There is increasing interest in oxidative stress being a potential aetiological factor and/or a triggering factor in Crohn's disease, rather than a concomitant occurrence during the pathogenesis of the disease. Recent research has shown that the immune mononuclear cells of Crohn's disease patients are induced to produce hydrogen peroxide (H2O2). Similarly, the regulation of antioxidant enzymes during disease in these cells has been unravelled, showing that SOD (superoxide dismutase) activity and GPx (glutathione peroxidase) activity is increased during active disease and returns to normal in remission phases. However, catalase remains constantly inhibited which supports the idea that catalase is not a redox-sensitive enzyme, but a regulator of cellular processes. ROS (reactive oxygen species) can be produced under the stimulus of different cytokines such as TNFα (tumour necrosis factor α). It has been shown in different experimental models that they are also able to regulate apoptosis and other cellular processes. The status of oxidative stress elements in Crohn's disease and their possible implications in regulating cellular processes are reviewed in the present paper.