Complication Of Chronic Inflammation Research Articles

Prognostic possibilities of CT in the diagnosis of chronic inflammatory complications of diverticular disease

Objective: to determine the diagnostic effectiveness of computed tomography (CT) in predicting the course of the disease in patients with chronic inflammatory complications of diverticular disease (DD).Material and methods. The study included 70 patients with a complicated course of colon diverticular disease in the phase of exacerbation of the chronic inflammatory process. All patients underwent CT of the abdominal cavity with intravenous contrast to assess the type and severity of inflammatory changes in the colon and surrounding tissues in the area of localization of diverticula. All patients received conservative treatment and were monitored as part of the ongoing study for 12 months from the moment of initial treatment at the Center. Surgical intervention due to the ineffectiveness of conservative therapy or the recurrence of the inflammatory process during the established follow-up period was considered as an unfavorable outcome of the disease (42/60%). The positive effect of drug therapy without signs of a return of the clinical picture of inflammation within 12 months was considered as a favorable outcome (28/40%). A statistical analysis of CT signs of inflammatory changes in various DD outcomes was performed to identify prognostic CT parameters.Results. Statistically significant differences were revealed between the severity of inflammatory changes according to CT data for different outcomes of the disease. In the favorable outcome group, the main part (23/28.82%) were patients with diverticulitis, in the unfavorable outcome group, 2/3 of patients (29/42.64%) were diagnosed with pericolic infiltrates, including abscesses/cavities, and colon fistulas. It was found that the thickness of the intestinal wall, the extent of inflammatory changes in the intestinal wall, the extent of inflammatory infiltration of pericolic fiber, the symptom of “centipede”, the accumulation of fluid in the pericolic region statistically significantly differed with different outcomes of chronic inflammatory complications of diverticular disease. Мultivariate Cox proportional hazard model revealed two main predictors of the onset of an unfavorable outcome – thickening of the intestinal wall and the presence of fluid in the pericolic region. Thickening of the intestinal wall at values equal to or greater than 0,6 cm increased the risk of an unfavorable outcome by 4.69 times, and the presence of fluid by 4.52 times.Conclusion. The use in clinical practice of the revealed CT predictors of the onset of an unfavorable outcome in chronic inflammatory complications of DB can serve as one of the factors for deciding on elective surgery in this category of patients.

Value of computed tomography and abdominal ultrasound for chronic inflammatory complications of diverticular disease

Aim: to evaluate the diagnostic value of computed tomography (CT) and abdominal ultrasound (US) for chronic inflammatory complications in patients with diverticular disease (DD).Patients and Methods: the prospective cohort study included 50 patients with complicated DD. All patients underwent preoperative abdominal CT with intravenous contrast and abdominal US, with further elective bowel resection. The results of CT and ultrasound were compared with morphology of the removed specimens.Results: the sensitivity and specificity for chronic diverticulitis was 66.7% and 95.7% for CT and 100.0% and 95.7% for US. For chronic pericolic abdominal mass it was 94.8% and 90.9% for CT, 94.8% and 100.0% for US; for abdominal abscesses/cavities it was 87.5% and 96.2% for CT and 91.6% and 100.0% for US; for diverticular fistulas it was 87.5% and 100.0% for CT and 87.5% and 100.0% for US. No significant differences were obtained between two diagnostic modalities. A high level of consistency (κ-coefficient 0.71) of CT and US for the diagnosis of inflammatory complications of DD was found. Conclusion: CT and US have a similar high diagnostic value for chronic inflammatory complications of DD. Each of them can be used as a single diagnostic modality or both depending on the clinical case.

Non-canonical function of histone methyltransferase G9a in the translational regulation of chronic inflammation

We report a novel translation-regulatory function of G9a, a histone methyltransferase and well-understood transcriptional repressor, in promoting hyperinflammation and lymphopenia; two hallmarks of endotoxin tolerance (ET)-associated chronic inflammatory complications. Using multiple approaches, we demonstrate that G9a interacts with multiple translation regulators during ET, particularly the N6-methyladenosine (m6A) RNA methyltransferase METTL3, to co-upregulate expression of certain m6A-modified mRNAs that encode immune-checkpoint and anti-inflammatory proteins. Mechanistically, G9a promotes m6A methyltransferase activity of METTL3 at translational/post-translational level by regulating its expression, its methylation, and its cytosolic localization during ET. Additionally, from a broader view extended from the G9a-METTL3-m6A translation regulatory axis, our translatome proteomics approach identified numerous "G9a-translated" proteins that unite the networks associated with inflammation dysregulation, Tcell dysfunction, and systemic cytokine response. In sum, we identified a previously unrecognized function of G9a in protein-specific translation that can be leveraged to treat ET-related chronic inflammatory diseases.

Oxidative stress in sickle cell anemia can be a prognostic marker for disease severity

CONTEXT: Sickle cell anemia (SCA) or sickle cell disease (SCD) is an inherited blood disorder characterized primarily by chronic anemia and periodic episodes of pain. There are reports that increase in oxidative stress may play a significant role in the pathophysiology of SCA. AIM: The present study aims to investigate enzymatic and nonenzymatic antioxidant status in SCA patients to understand the incidence of increased oxidative stress in the populace of tribal Palghar region of Western Maharashtra. SUBJECTS AND METHODS: Patients with SCA (n = 250) and age- and sex-matched healthy persons (n = 250) as controls from the primary health center of Palghar were included in this study. Informed written consent was obtained from all the participants. RESULTS: Activities of enzymatic antioxidants such as glutathione peroxidase, glutathione-S-trasferase, catalase, and level of nonenzymatic antioxidants such as glutathione, Vitamin E and C decreased significantly in SCA participants when compared to controls. The level of lipid peroxides and activity of superoxide dismutase increased significantly above normal in SCA participants. SCA is characterized by the increased generation of reactive-oxygen species resulting in oxidative damage of various cell types, including erythrocytes and this chronically elevated oxidative stress in SCD might play a significant role in the increased autoxidation of Sickle hemoglobin (HbS), increased intravascular hemolysis, ischemia-reperfusion injury, and chronic inflammatory complications. CONCLUSION: The present study indicates that oxidative stress can be considered one of the prognostic markers to evaluate the clinical severity of the SCA participants.

O10 Multi-disciplinary working in SpA diagnosis: case study

Case report - IntroductionHigh-intensity exercise is effective in treating Axial Spondyloarthritis (SpA). A reduction in morbidity therefore should be gained from accurate diagnosis early in the disease history. In 2021 the National Axial Spondylarthritis Society (NASS) released a statement to reduce time to diagnosis (TTD) of Axial SpA from 8 years to 1, to reduce morbidity and long-term complications of chronic inflammation.This case study aims to provide an example of how a multidisciplinary team working in close clinical proximity (CCP MDT) can be utilised to reduce the TTD and optimise treatment and biomechanics of acute and chronic SpA.Case report - Case descriptionCases 1 and 2 compare patients presenting to a musculoskeletal clinic with lower back pain, stiffness, and reduced range of movement. Case 1. Subacute presentation: 25-year-old male with a 3-month history of lower back pain and stiffness. This progressed to a month of not being able to walk, disturbed sleep and morning stiffness of > 1hr. Clinical examination showed no extra musculoskeletal manifestations, and no other joint pain. Tests showed HLA B27 positive status with florid bilateral sacroiliitis on MRI.Case 2. Chronic presentation: 34-year-old male with 20-year history of insidious onset lower back pain previously diagnosed as “non-specific” back pain by a series of health care professionals (HCPs). HLA B27 negative, MRI showed acute on chronic SIJ arthropathy with juxtarticular sclerosis.Patients were seen by a HCP who referred to a rheumatologist. In both, clinical and radiological signs indicated Axial SpA which was treated with 120mg of IM Depo-Medorone and 8-weeks of strength-based active intervention from a physiotherapist and strength and conditioning coach. BASDAI, 5-rep max single leg press scores were recorded pre- and post-intervention (Table 1.). Case 1Pre-interventionPost-interventionBASDAI5.10.9Single leg press 5RM55kg75kgCase 2Pre-interventionPost-interventionBASDAI 6.14.2Single leg press 5RM65kg115kgCase report - DiscussionConventional therapies for SpA back pain are effective even in chronic cases; this is reflected in the improvements made across both cases. Using a similar approach of high-intensity exercise and mobility both were able to significantly improve BASDAI scores and lower limb strength.We can hypothesise that attempts to innovate the biomechanics of the treatments is missing the fundamental issue surrounding SpA back pain highlighted by NASS; TTD takes too long for too many. There is a feasible argument therefore that it is the structures of how we operate as HCPs that is reducing the efficacy of said treatments. This is evidenced by case 2 (Table 1), whose multiple presentations to different HCPs over 20 years led to potentially preventable prolonged morbidity as well as increased the risk of future complications due to chronic inflammation. This is a story that is sadly reflective of the disease histories of many SpA patients.Much like any primary care provider, patients at this clinic will first present to a HCP. The difference is that all HCPs are trained to recognise features of SpA back pain and encouraged to flag concerns to a doctor.HCPs work in adjacent spaces allowing for better sharing of ideas and concerns that is often reduced by physical separation. This creates the concept of a CCP MDT. HCPs are encouraged to share concerns face to face without reproach or the formality or delay of a written request. This structure has proven to be effective in diagnosis in both of these cases but has proven to be as useful in many more instances to rule out a differential or allay a concern.These cases argue that the low lying fruit of improving clinical outcomes in this cohort is addressing how we as HCPs interact and cooperate. Case report - Key learning pointsBiomechanics of the treatments of SpA are effective in the majority of cases when applied by sufficiently skilled practitioners. It is the TTD that we should address to aid the reduction in morbidity in this patient group through the following recommendations: Foster education of SpA in allied health professionals. This allows us to be aware of the boundaries of our own knowledge and at what point a different clinical perspective should be sought.Diversify clinical environments and encourage MDT working. The use of CCP MDT structures allows more nuanced access to different opinions in a safe supportive manner, allowing for sharing of ideas and concerns – allowing for supporting diverse expertise and insight. If we are to reduce TTD to the 1-year NASS hopes for, we should first look at how we work, not what we do. Ongoing research and auditing of patients is being carried out to better assess how educational practices can be optimised to help meet the NASS targets. For more information or to collaborate contact matthew.virden@puresportsmed.com

Abstract 753: Associations of inflammatory bowel disease and subsequent cancers in the United States elderly population

Abstract Introduction: The risk for cancer may be elevated in patients with inflammatory bowel disease (IBD) due to chronic inflammation caused by IBD and/or immunosuppressive treatments. A comprehensive investigation of cancer as a complication of IBD in elderly patients is needed, as this population has a high background risk of cancer and are underrepresented in IBD studies. Methods: We performed a case-control study using Surveillance Epidemiology and End Results-Medicare linked data. Cancer cases (N=1,986,735) included all patients aged 66-99 with a first invasive neoplasm diagnosed in 1992-2015. Cancer-free controls (N=200,000) were randomly selected from Medicare and matched by sex, age, and selection year. Medicare claims containing ulcerative colitis (UC) or Crohn's disease (CD) diagnosis codes occurring at least one year prior to selection were used to identify IBD patients. We estimated odds ratios (OR) and 95% confidence intervals (CI) with logistic regression, adjusting for the matching factors, race, and the average number of physician visits per year. A Bonferroni-corrected threshold of significance was defined as p < 0.0003 for 156 tests. Colorectal cancer analyses were further adjusted for the average number of colorectal cancer screens per year occurring at least one year before selection. Results: IBD, CD, and UC were present in 0.80%, 0.33%, and 0.53% of cancer cases and in 0.76%, 0.30%, and 0.52% of controls, respectively. Out of 52 cancers examined, IBD was significantly associated with cancers of the small intestine (OR=2.53; 95% CI=2.14-2.99), intrahepatic (1.93; 1.48-2.52) and extrahepatic bile ducts (1.73; 1.37-2.19), rectum (1.67; 1.43-1.94), colon (1.20; 1.10-1.31), and lung (1.14; 1.07-1.23). For hematological malignancies, IBD was significantly associated with myelodysplastic syndrome (MDS) (OR=1.45; 95% CI=1.28-1.65) and diffuse large B-cell lymphoma (DLBCL) (1.28; 1.13-1.46). An inverse association was observed for IBD and cancer of the uterus (0.72; 0.62-0.83). CD was associated with cancers of the small intestine (OR= 4.21; 95% CI= 3.40-5.22), myelodysplastic syndrome (1.74; 1.44-2.11), rectum (1.60; 1.1.26-2.03), and lung (1.29; 1.16-1.43) and. UC was associated with cancers of the gastrointestinal system: intrahepatic bile ducts (1.94; 1.41-2.67), rectum (1.88; 1.57-2.25), and colon (1.38; 1.25-1.53). Conclusions: Our large population-based analysis evaluating a comprehensive range of cancers found IBD significantly associated with increased risk for cancers of the small intestine, biliary tract, colorectum, and lung. IBD was also associated with DLBCL and MDS. Positive associations with IBD and cancers of the gastrointestinal system may reflect complications of chronic inflammation, while associations with DLBCL and MDS may reflect the effect of immunosuppressive medications. Citation Format: Jeanny Wang, Monica D'Arcy, Eric Engels, Minkyo Song. Associations of inflammatory bowel disease and subsequent cancers in the United States elderly population [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 753.

Ultrasound diagnosis of chronic paracolic inflammatory mass in diverticular disease

To evaluate the feasibility of ultrasound in diagnosis of chronic paracolic inflammatory mass in patients with diverticular disease. We analyzed ultrasonic findings in 216 patients with chronic inflammatory complications of colonic diverticular disease. Chronic paracolic inflammatory mass as the most common and significant chronic complication of diverticular disease was analyzed in 116 patients. Ultrasonic findings were compared with specimen assessment, intraoperative data, irrigoscopy, colonoscopy, endoscopic ultrasound and computed tomography data. Sensitivity of ultrasound for diagnosis of chronic paracolic inflammatory mass was 76,7%, specificity - 100%, overall accuracy - 87,5%. CT and endoscopic ultrasound were the most informative among different diagnostic tools (sensitivity 79,6% and 77,8%, respectively). Ultrasonic examination and computed tomography are the most valuable methods for diagnosis of chronic paracolic inflammatory mass in patients with diverticular disease. Ultrasound is a first-line method for diagnosis and follow-up of complicated diverticular disease due to its availability, safety and unnecessary special preparation of patients.

One - stage laparoscopic treatment of chronic inflammatory complications of diverticular colon disease

Introduction. Diverticulosis of the colon is one of the most common diseases of the intestine. In recent years, there has been an increasing tendency for diverticular disease (DD) to manifest itself at earlier age, as well as to the more frequent development of complications in young and middle - aged people. Most interventions for planned surgical treatment of DD complications are performed by two - stage access for laparotomy. One - stage laparoscopic intervention is a relatively new approach to treatment. Case report. 40-year - old man was hospitalized in the surgical department with complaints of air release during urination and aching pain in the lower part of abdomen. Outpatient examination excluded the pathology of the prostate gland and revealed infiltration between the urinary bladder and sigmoid colon. A complex examination, including cystoscopy, colonoscopy, multispiral computed tomography with contrast, showed the presence of chronic paracolic infiltration and blad - derntestinal fistula, which complicated the latent course of colon DD. Laparoscopic opening the abscess, resection of the sigmoid colon, the formation of descendo - rectal anastomosis was performed in one - step. Intra - and postoperative complications were not observed, dynamic observation and control examination after 6 months showed no symptoms. Discussion. One - stage laparoscopic surgery in the treatment of chronic inflammatory complications of colon DD is currently available procedure, which allows in a short time to relieve patients from symptoms and has good immediate and long - term results. The condition for the use of this technique is a sufficient experience of laparoscopic surgery of the abdominal cavity, retroperitoneal space and pelvis.

Regulation of FOXP3 Expression by c-Rel O-GlcNAcylation

Abstract The NF-kappaB protein c-Rel plays a critical role in controlling autoimmunity through regulating T cell function and T regulatory cell development. Type 1 diabetes is a T cell-mediated autoimmune disease associated with hyperglycemia and chronic inflammatory complications. Previously, we discovered that hyperglycemia induces hyper O-GlcNAcylation of c-Rel at serine residue 350 and enhances the transcription of c-Rel-dependent pro-autoimmune cytokines interleukin-2 (IL-2), interferon gamma (IFNG) and granulocyte macrophage colony-stimulating factor (GM-CSF). In contrast, our recent results show that c-Rel O-GlcNAcylation decreases forkhead boxP3 (FOXP3) expression in T cells. Chemically enhancing cellular O-GlcNAcylation decreases the DNA binding ability of c-Rel at the FOXP3 promoter as demonstrated both by oligonucleotide pulldown and chromatin immunoprecipitation assays. Moreover, mutation of serine 350 in c-Rel to alanine augments T cell receptor-induced FOXP3 expression. O-GlcNAcylation-dependent suppression of FOXP3 was also found in vivo in streptozotocin-induced diabetic mouse model. This study reveals a novel molecular mechanism that regulates hyperglycemia-dependent c-Rel function in autoimmune diabetes, with potential to develop novel therapeutics targeting c-Rel O-GlcNAcylation.

Tattoos and skin barrier function: Measurements of TEWL, stratum corneum conductance and capacitance, pH, and filaggrin.

Initially after tattooing, the skin barrier function is broken. However, the long-term impact of clinically healed tattoos on this has never been studied. The aim was to investigate the long-term effect on the skin barrier function in normal tattoos and examples of tattoos with chronic inflammatory complication. Participants were recruited from the "Tattoo clinic" of the Dermatological Department on Bispebjerg Hospital in Denmark, where patients with complicated tattoo reactions are treated. Transepidermal water loss (TEWL), conductance, capacitance, and pH were measured in tattooed skin with regional control measurements in normal non-tattooed skin. Natural moisturizing factor (NMF) was measured in collected tape strips. Twenty six individuals with 28 tattoos were included, that is, 23 normal tattoos without any pathologic reaction and 5 tattoos with chronic inflammatory complications. No significant differences were found in tattooed versus non-tattooed skin with respect to TEWL (median values 6.6 vs 7.2g/m2 /h), conductance (76 vs 78a.u.), pH (5.94 vs 5.79), and NMF (0.58 vs 0.59mmol/g protein). Capacitance (64 vs 57a.u.) was higher in tattooed skin compared to non-tattooed skin (P=0.006). Similar results were found in tattoos with inflammatory reactions. Overall, skin tattoos do not affect the long-term skin barrier function markedly. The skin capacitance was, however, affected in tattooed skin areas compared to non-tattooed skin areas.

Prevalence and clinical challenges among adults with primary immunodeficiency and recombination-activating gene deficiency

Prevalence and clinical challenges among adults with primary immunodeficiency and recombination-activating gene deficiency

Changing epidemiology of AA amyloidosis: clinical observations over 25 years at a single national referral centre

Objective: Systemic AA amyloidosis is a serious complication of chronic inflammation; however, there are relatively few published data on its incidence. We investigated the changing epidemiology of AA amyloidosis over a 25-year period at a single national referral centre.Methods: We conducted a retrospective study of all patients diagnosed with AA amyloidosis who had attended the centre between 1990 and 2014 inclusive. Six hundred and twenty-five patients were studied in three cohorts: C1: 1990–1997; C2: 1998–2006; C3: 2007–2014.Results: Mean age at presentation increased from 46 in C1 to 56 in C3 (p < .0001). The proportion of South Asian patients increased from 4% in C1 to 17% in C3 (p = .0006). Comparison of underlying diseases between C1 and C3 revealed a reduction in patients with juvenile idiopathic arthritis from 25% to 2% (p < .0001), but an increase in patients with chronic infection due to intravenous recreational drug use from 1% to 13% (p < .0001), and uncharacterized inflammatory disorders from 10% to 27% (p <.0001). More patients were in end-stage renal failure at presentation in C3 (29%) than C1 (15%) (p = .0028). Median age at death was later in C3 (62 years) than C1 (54 years) (p = .0012).Conclusion: These data suggest both falling incidence and better outcome in AA amyloidosis over a quarter of a century, reflecting advances in therapeutics and overall management of complex chronic disease in an ageing population. AA amyloidosis of uncertain aetiology presents an emerging major problem. Newer techniques such as next-generation sequencing may aid diagnosis and effective treatment, thereby improving overall survival.

Characterizing the Inflammasome in Polycythemia Vera

Polycythemia vera (PV) is characterized by the presence of the Janus kinase 2 (JAK2) V617F mutation in 97% of patients. This results in constitutive activation of the JAK/signal transduction and activators of transcription (STAT) pathway. JAK2 can also enter the nucleus directly following SUMOylation to effect alterations in histone function. Patients with PV display elevated peripheral blood counts, chronic inflammation and cytokine driven symptoms leading to morbidity and reduced quality of life. An important target of inflammation via JAK-mediated interferon (IFN) signalling is promyelocytic leukemia protein (PML), necessary for nuclear body (NB) formation. NBs act as a hub for transcriptional regulation and histone modification, bringing together multiple proteins to elicit diverse cellular functions including apoptosis, cellular senescence, DNA repair and inflammatory responses.Multiplex analysis of a 29 cytokine/chemokine panel in normal donor (n=8) and PV patient (n=10) serum, indicated significant upregulation of inflammatory cytokines through both JAK2-dependent and independent signalling, including IFNγ, interleukin (IL)-5, IL-12 (p< 0.05; JAK2 dependent), and CXCL10, MIP-1α and TNF-α (p< 0.001; JAK2 independent). PV patients also had significant deregulation of JAK2 dependent and IFN response genes (n=84 genes) especially upregulation of GBP1 (p< 0.05), MGST3 and SUMO3 (p< 0.001) and downregulation of PML (p< 0.01) and NFκB (p< 0.001). Using Duolink® in situ proximity ligation assays in both JAK2 V617F cell lines (UKE1 and SET2) and PV patient/normal donor monocytes, we demonstrate that JAK2 is SUMOylated and actively co-localises with PML in the cell nucleus. Importantly, there was a significant increase in the degree of interaction between JAK2 and PML in PV monocytes compared to normal donors (n=3, p< 0.001). Treatment of UKE1 and SET2 cell lines with the JAK1/2 inhibitor ruxolitinib led to a decrease in JAK2 dependent and IFN response genes as well as the degree of JAK2/PML co-localisation indicating active JAK2 mediates these responses.To determine whether the deregulation of these cellular processes is manifested by a functional alteration in the myeloid lineages of patients with PV, we analysed their monocyte profiles and the ability of monocytes to form M1 and M2 macrophages as well as neutrophil function. Monocytes exist in different subsets defined by the expression of CD14 and CD16. In PV, the proinflammatory intermediate subset was significantly increased (16.4% ±2.04 vs 6.5% ±1.08, p< 0.05, n=5) at the expense of the classical subset of monocytes (79% ±2.04 vs 88.9% ±1.07, p< 0.05, n=5). The non-classical, IL-1RA producing subset remains unchanged (3.6% ±0.38 vs 4.6% ±0.48 p= 0.1). Macrophages generated from these monocytes were polarized for M1 or M2 differentiation using GM-CSF and M-CSF respectively. They were then cultured in the presence of normal or PV patient serum with or without stimulation; IFNγ and lipopolysaccharide (LPS) for M1 macrophages and IL-4 for M2 macrophages. Culture supernatant was then assessed for the same 29 cytokine/chemokine panel as above and showed significant alterations including an increase in IL-6 and CXCL10 in PV derived M1 macrophages (p< 0.001), while M2 macrophages had a decrease in CXCL1 and increase in IL-1RA (p< 0.01).Neutrophils were cultured for 24 hours in the presence of normal or PV patient serum with or without stimulation with LPS and showed significant alteration in the production of several proinflammatory cytokines/chemokines compared to neutrophils cultured in Promocell Base Media DXF alone including IFNγ, VEGF, and CXCL10 (p< 0.001), indicating that serum from PV patients can elicit neutrophil activation.In conclusion, we have shown that inflammation is a major pathophysiological process in PV patients resulting in increased PML/JAK2 co-localisation, increased transcription of direct JAK2 target and IFN response genes, high circulating proinflammatory cytokine/chemokine levels, altered proinflammatory monocyte profiles, and significant change in the secretory production of mature granulocytes following stimulation. Treatment of JAK2 V617F positive cell lines with ruxolitinib significantly alters the inflammasome supporting its efficacy in the treatment of PV patients to alleviate the symptoms and complications of chronic inflammation. DisclosuresDrummond:Gilead: Membership on an entity's Board of Directors or advisory committees; Celgene: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Novartis: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Baxalta: Membership on an entity's Board of Directors or advisory committees. Copland:BMS: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Ariad: Honoraria, Membership on an entity's Board of Directors or advisory committees; Pfizer: Honoraria, Membership on an entity's Board of Directors or advisory committees.

Abstract 3165: Galectin-4, a key regulator of inflammation and cancer of the intestine

Abstract The inflammatory bowel diseases including ulcerative colitis and Crohn's disease are chronic inflammatory disorders of the intestine. It is well recognized that a serious long-term complication of chronic inflammation is the development of colorectal cancer (CRC). Although various mechanisms underlying different chronic inflammation conditions are known, it is increasing clear that proinflammatory cytokines produced during immune responses are the key contributing factors to the above disease states. On the other hand, genetic alterations inducing CRC also promote inflammatory responses in the gastrointestinal microenvironment. This interdependent relationship between oncogenesis and inflammation suggests that it is possible to identify a regulatory molecule that links these two cellular processes. The long-term interest of our efforts is to determine the mechanisms of galectin-4 (gal-4) in the normal and disease physiology of the human gastrointestinal tract. Gal-4 is expressed predominantly by the gastrointestinal epithelia. It is secreted and binds to specific oligosaccharide moiety in glycoproteins on the cell surface. We have recently demonstrated that gal-4 down-regulates cell proliferation and its loss of expression coincides with CRC onset and progression, and suggested that gal-4 functions as a tumor suppressor. The purpose of this study is to determine whether gal-4 regulates inflammatory responses in colorectal cells. In this study, we used gal-4 +ve CRC cells, HT-29 and LS-180 and neutralized their surface-bound gal-4 with gal-4 antibodies. Such cells proliferated profusely with concomitant up-regulation of transcription of 47 genes of which 17 are known partners/gene targets of multiple inflammation signaling pathways. Extracellular (spent) media obtained from these experiments were found to contain unique cytokines that are known to be pro-tumorigenic. In converse experiments, exposure of gal-4 -ve HCT116 cells to purified gal-4 resulted in decreased cell proliferation. Concomitantly, there was increased expression of p27, decreased expressions of cyclin D1 and c-Myc. These data suggested that binding of gal-4 to cell surface arrests cell cycle. Finally, analysis of rat colon tissue sections obtained from dextran sulfate-sodium (DSS) induced chronic colitis (kindly given by Dr. Daniel W. Rosenberg, Univ. of Connecticut CT) suggested that the epithelial cells at the colitis regions exhibited loss of gal-4 expression. These data together suggested that surface gal-4 down-regulates inflammatory response pathways, cell proliferation and oncogenic events. These data also suggested that gal-4 protein therapy and re-expression of gal-4 in lesions of chronic inflammation and CRC are potentially attractive methods of controlling these diseases. *A part of this study was carried out by the authors while working at the Texas Tech University Health Sciences Center Note: This abstract was not presented at the meeting. Citation Format: U.S. Rao, Prema S. Rao. Galectin-4, a key regulator of inflammation and cancer of the intestine. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3165. doi:10.1158/1538-7445.AM2015-3165

Can HIV Be Cured and Should We Try?

An estimated 35.0 million persons live with HIV; 2.1 million new infections occurred and 1.5 million persons died of HIV in 2013 (World Health Organization, http://www.who.int/hiv/data/epi_core_dec2014.png?ua=1). Despite effective combination antiretroviral therapy (cART), less than one-quarter of patients can access these life-prolonging medications; and despite its effectiveness, cART does not normalize life expectancy, as premature aging, metabolic complications and chronic inflammation complicate HIV therapy. HIV is incurable due to the presence of a latent viral reservoir. During the life cycle of the virus, HIV integrates into the host DNA. A subset of integrated HIV provirus remains transcriptionally silent, producing neither viral proteins nor viral progeny, until reactivation by various physiologic stimuli. This latency of HIV allows some infected cells to escape immune detection and elimination, and these latently infected cells constitute the viral reservoir. The latent viral reservoir allows viral rebound within weeks of interruption of cART, 1,2 where the magnitude of viral replication approaches that present pre-therapy. Although it was once thought that viral rebound occurred universally following therapy interruption, several recent reports challenge that paradigm. The “Berlin patient” successfully cleared HIV after two allogeneic transplants from a donor with homozygous CCR5Δ32 mutation,3 and he has not rebounded HIV after nearly seven years. This case likely represents a cure from HIV; yet other cases have been described where HIV rebound has been attenuated, or delayed. The “Mississippi” baby was a perinatally HIV-infected infant who initiated cART within hours of birth, and when interrupted eighteen months later, viremia remained undetectable for nearly two years.4 The Harvard BMT cases underwent allogeneic BMT, developed undetectable HIV DNA, yet rebounded viremia within only eight months after cART discontinuation.5 Together these cases demonstrate that control of viremia in the absence of cART is possible, if not durable.4 The VISCONTI cohort of 14 HIV-infected adults who initiated antiretroviral therapy during acute infection, and in whom high level rebound viremia had not occurred several years after cessation of therapy,6 similarly demonstrated that viral rebound following cART interruption can be attenuated. These cases have reinvigorated research toward finding a cure for HIV, which certainly will require an exceptional investment of time, talent and resources. Thus it is prudent to question whether such resources would be better devoted to more proximate needs with proven results (such as supplying cART to those without access to it, or supplying pre/post exposure prophylaxis to reduce the rate of new infections).

Molecular regulation of autoimmunity by O-GlcNAcylation of NF-kappaB c-Rel in T lymphocytes (IRM11P.633)

Abstract The NF-kappaB protein c-Rel plays a critical role in controlling autoimmunity through regulating T cell function and T regulatory cell development. Type 1 diabetes is an autoimmune disease associated with hyperglycemia and chronic inflammatory complications. We found that hyperglycemia induces O-GlcNAcylation of c-Rel at serine residue 350 and activates the transcription of c-Rel-dependent pro-autoimmune cytokines interleukin-2 (IL-2), interferon gamma (IFNG) and granulocyte macrophage colony-stimulating factor (GM-CSF). Our recent results show that c-Rel O-GlcNAcylation decreases FOXP3 expression in T cells. These results suggest that the glucose metabolism-dependent c-Rel O-GlcNAcylation serves as a key regulatory switch with dual roles in controlling T cell and T regulatory cell function in autoimmune diabetes. Despite decades of research, drugs based on molecular targets to treat type 1 diabetes have remained elusive. This study reveals a novel molecular mechanism that regulates NF-kappaB c-Rel and autoimmune diabetes, with potential to develop novel therapeutics targeting c-Rel O-GlcNAcylation. Moreover, this study serves as the basis to explore the role of c-Rel O-GlcNAcylation in other autoimmune diseases such as celiac disease, lupus and arthritis, where c-Rel function have been implicated.

Inflammatory Macrophages Promotes Development of Diabetic Encephalopathy

Background/Aims: Diabetes and Alzheimer's disease are often associated with each other, whereas the relationship between two diseases is ill-defined. Although hyperglycemia during diabetes is a major cause of encephalopathy, diabetes may also cause chronic inflammatory complications including peripheral neuropathy. Hence the role and the characteristics of inflammatory macrophages in the development of diabetic encephalopathy need to be clarified. Methods: Diabetes were induced in mice by i.p. injection of streptozotocin (STZ). Two weeks after STZ injection and confirmation of development of diabetes, inflammatory macrophages were eliminated by i.p. injection of 20µg saporin-conjugated antibody against a macrophage surface marker CD11b (saporin-CD11b) twice per week, while a STZ-treated group received injection of rat IgG of same frequency as a control. The effects of macrophage depletion on brain degradation markers, brain malondialdehyde (MDA), catalase, superoxidase anion-positive cells and nitric oxide (NO) were measured. Results: Saporin-CD11b significantly reduced inflammatory macrophages in brain, without affecting mouse blood glucose, serum insulin, glucose responses and beta cell mass. However, reduced brain macrophages significantly inhibited the STZ-induced decreases in brain MDA, catalase and superoxidase anion-positive cells, and the STZ-induced decreases in brain NO. Conclusion: Inflammatory macrophages may promote development of diabetic encephalopathy.

Risk factors for subclinical inflammation in children with familial mediterranean fever

Familial Mediterranean fever (FMF) is the most common autosomal-recessive inherited inflammatory disease characterized by attacks of painful inflammation. Some patients with FMF have subclinical inflammation persisting between the attacks. Persistent inflammation may lead to amyloidosis and other complications of chronic inflammation.

Pathogenetic mechanisms of amyloid A amyloidosis

Systemic amyloid A (AA) amyloidosis is a serious complication of chronic inflammation. Serum AA protein (SAA), an acute phase plasma protein, is deposited extracellularly as insoluble amyloid fibrils that damage tissue structure and function. Clinical AA amyloidosis is typically preceded by many years of active inflammation before presenting, most commonly with renal involvement. Using dose-dependent, doxycycline-inducible transgenic expression of SAA in mice, we show that AA amyloid deposition can occur independently of inflammation and that the time before amyloid deposition is determined by the circulating SAA concentration. High level SAA expression induced amyloidosis in all mice after a short, slightly variable delay. SAA was rapidly incorporated into amyloid, acutely reducing circulating SAA concentrations by up to 90%. Prolonged modest SAA overexpression occasionally produced amyloidosis after long delays and primed most mice for explosive amyloidosis when SAA production subsequently increased. Endogenous priming and bulk amyloid deposition are thus separable events, each sensitive to plasma SAA concentration. Amyloid deposits slowly regressed with restoration of normal SAA production after doxycycline withdrawal. Reinduction of SAA overproduction revealed that, following amyloid regression, all mice were primed, especially for rapid glomerular amyloid deposition leading to renal failure, closely resembling the rapid onset of renal failure in clinical AA amyloidosis following acute exacerbation of inflammation. Clinical AA amyloidosis rarely involves the heart, but amyloidotic SAA transgenic mice consistently had minor cardiac amyloid deposits, enabling us to extend to the heart the demonstrable efficacy of our unique antibody therapy for elimination of visceral amyloid.

Mo1823 Involvement of MDR1 in Intestinal Epithelial Injury Caused by Acetylsalicylic Acid

Introduction Intestinal fibrosis (IF) is a major complication of chronic inflammation of the intestinal tissue in inflammatory bowel disease (IBD). IF can cause narrowing of the intestinal lumen, which may lead to stricture formation. Currently, adequate models are lacking to study the mechanism of intestinal fibrosis in IBD. Our aim is to develop an ex vivo model for IF by using mouse precision-cut intestinal slice (PCIS). In addition, regional differences in the onset of fibrosis in the bowel will be studied. In PCIS all cell types are present in their original tissue-matrix environment. As fibrosis is a multicellular phenomenon, PCIS can be used as a model to study the early onset of intestinal fibrosis. Methods Mouse jejunum, ileum and colon were excised and prepared as segments embedded in agarose. PCIS (estimated 300-400 μm) were prepared and incubated up to 48 hr. ATP content of the PCIS was used to assess the general viability. To study the early onset of fibrosis, the gene expression of different fibrosis markers: Pro-Collagen 1 A1 (COL1A1), Heat Shock Protein 47 (HSP47), alpha-Smooth Muscle Actin (SMA) and Fibronectin (FN1) was determined in PCIS after 48 hr of incubation. Results Mouse PCIS from the different regions of the bowel were viable up to 48 hr in culture. After 48 hr of incubation of PCIS, HSP47 and FN1 gene expression was significantly up-regulated, compared to PCIS directly after slicing, in jejunum (3.9 and 3.6 fold, respectively) and in ileum (4.5 and 4.9 fold, respectively). Gene expression of Col1A1 (0.7 fold) was only down-regulated in PCIS from the colon. SMA gene expression was down-regulated in PCIS from all the regions of the intestine. PCIS from jejunum cultured in the presence of 5ng/mL Transforming-Growth Factor (TGF)β1 for 48 hr, HSP47 (1.2 fold), FN1 (4.6 fold) and COL1A1 (2.0 fold) were significantly upregulated compared to control PCIS after 48 hr of incubation. In addition, in PCIS from Ileum incubated with 5ng/mL TGF-β1, the gene expression of HSP47 (1.9 fold) and FN1 (3.9 fold) was significantly increased. In PCIS from the colon cultured for 48 hr in the presence of 5ng/ml TGF-β1, the gene expression of fibrosis markers was not affected. Conclusion Mouse PCIS from different regions of the bowel can successfully be cultured for 48 hr. Fibrosis markers are induced by incubation up to 48 hr and by addition of TGFβ1 in jejunum and ileum PCIS, but not in colon PCIS. We are the first to show that regional differences in the onset of intestinal fibrosis can be measured using PCIS.