A novel unconventional T cell population enriched in Crohn's disease.

OBJECTIVE: One of the current hypotheses to explain the proinflammatory immune response in IBD is a dysregulated T cell reaction to yet unknown intestinal antigens. As such, it may be possible to identify disease-associated T cell clonotypes by analysing the peripheral and intestinal T-cell receptor (TCR) repertoire of patients with IBD and controls. DESIGN: We performed bulk TCR repertoire profiling of both the TCR alpha and beta chains using high-throughput sequencing in peripheral blood samples of a total of 244 patients with IBD and healthy controls as well as from matched blood and intestinal tissue of 59 patients with IBD and disease controls. We further characterised specific T cell clonotypes via single-cell RNAseq. RESULTS: We identified a group of clonotypes, characterised by semi-invariant TCR alpha chains, to be significantly enriched in the blood of patients with Crohn's disease (CD) and particularly expanded in the CD8+ T cell population. Single-cell RNAseq data showed an innate-like phenotype of these cells, with a comparable gene expression to unconventional T cells such as mucosal associated invariant T and natural killer T (NKT) cells, but with distinct TCRs. CONCLUSIONS: We identified and characterised a subpopulation of unconventional Crohn-associated invariant T (CAIT) cells. Multiple evidence suggests these cells to be part of the NKT type II population. The potential implications of this population for CD or a subset thereof remain to be elucidated, and the immunophenotype and antigen reactivity of CAIT cells need further investigations in future studies.

Changes in the inflammatory markers with advancing stages of diabetic nephropathy and the role of pentraxin-3.

BACKGROUND: Immunological and inflammatory mechanisms have been shown to have role in both the development and progression of diabetic nephropathy (DNP). There is need for more specific markers for inflammation as the ones commonly used are influenced by many factors. Pentraxin-3 (PTX-3) seems to be a potential candidate. We aimed in our study to evaluate the changes of PTX-3 levels in different stages of DNP and its relationship with other inflammatory markers. METHODS: This is a cross sectional study in which patients with DNP at different stages were involved. Patient were divided into three groups according to estimated glomerular filtration rate (eGFR), microalbuminuria and proteinuria levels: Group-1: eGFR >60 mL/min and microalbuminuria, Group-2: eGFR >60 mL/min and macroalbuminuria, Group-3: eGFR <60 mL/min and macroalbuminuria. Besides the routine biochemical parameters, levels of PTX-3, high sensitivity C-reactive protein (hsCRP), interleukin (IL)-1 and tumor necrosis factor (TNF)-α was measured. Groups were compared with each other regarding the study parameters and correlation of PTX-3 with other markers was evaluated. RESULTS: The mean PTX-3 level in Group-2 (0.94 ± 0.26 ng/mL) and -3 (1.35 ± 1.55 ng/mL) were higher than in Group-1 (0.81 ± 0.25 ng/mL) (p = 0.009 and p = 0.012). There was a significant correlation of PTX-3 with proteinuria (r = 0.266, p = 0.016), microalbuminuria (r = 0.304, p = 0.014) and hypoalbuminemia (r = 0.197, p = 0.043). PTX-3 was not correlated with other markers of inflammation (IL-1, TNF-α and hsCRP) and diabetic metabolic parameters (hbA1c, C-peptide, insulin and HOMA-IR). PTX-3, IL-1 and TNF-α levels increased with the advancing stage of DNP while hsCRP level did not change. CONCLUSION: PTX-3 that increases similar to other markers of inflammation (IL-1, TNF-α) is a better inflammatory marker than hsCRP. Furthermore, there is a relationship between PTX-3 and proteinuria independent from eGFR.

Interobserver Reliability of the Nancy Index for Ulcerative Colitis: An Assessment of the Practicability and Ease of Use in a Single-Centre Real-World Setting.

BACKGROUND: Histological disease severity assessment in ulcerative colitis [UC] has become a mainstay in the definition of clinical endpoints ['histological remission'] in clinical trials of UC. Several scores have been established in the microscopic assessment of disease activity, but the Nancy index [NI] stands out as being the histological index with the fewest scoring items. To what extent histological assessment using the NI is affected by interobserver reliability in a real-word setting is poorly understood. We therefore performed a single-centre retrospective analysis of NI assessment in patients with UC. METHODS: We retrospectively evaluated the NI in two independent cohorts [total: 1085 biopsies, 547 UC patients] of clinically diagnosed UC patients, who underwent colonoscopy between 2007 and 2020. Cohort #1 consisted of 637 biopsies from 312 patients, while Cohort #2 consisted of 448 biopsies from 235 patients. Two blinded pathologists with different levels of expertise scored all biopsies from each cohort. A consensus conference was held for cases with discrepant scoring results. Finally, an overall consensus scoring was obtained from both cohorts. RESULTS: The interobserver agreement of the NI was substantial after the assessment of 1085 biopsy samples (κ = 0.796 [95% confidence interval, CI: 0.771-0.820]). An improvement of the interobserver agreement was found with increasing numbers of samples evaluated by both observers (Cohort #1: κ = 0.772 [95% CI: 0.739-0.805]; Cohort #2: κ = 0.829 [95% CI: 0.793-0.864]). Interobserver discordance was highest in NI grade 1 [observer 1: n = 128; observer 2: n = 236]. Interobserver discordance was lowest in NI grades 0 [observer 1: n = 504; observer 2: n = 479] and 3 [observer 1: n = 71; observer 2: n = 66]. CONCLUSION: The NI is an easy-to-use index with high interobserver reliability for assessment of the histological disease activity of UC patients in a real-world setting. While NI grades 0 and 3 had a high level of agreement between observers, NI grade 1 had a poorer level of agreement. This highlights the clinical need to specify histological characteristics leading to NI grade 1.

Effects of alpha-tocopherol on acute pancreatitis in rat.

BACKGROUND: Acute pancreatitis is a disease with high morbidity and mortality, despite all the advances in technology. The overall mortality rate of acute pancreatitis is 10%, whereas the mortality rate in infected necrotizing pancreatitis is approximately 35%. In this study, we aimed to establish acute pancreatitis in rats in order to try out the alpha-tocopherol treatment protocol and to reveal the results biochemically and histopathologically. METHODS: Twenty-four male male Sprague-Dawley rats weighing between 300 and 350 g were used in the study. In Group 1, 80 µg/kg of normal saline was subcutaneously injected into eight rats; in Group 2, 80 µg/kg of cerulein was subcutaneously injected into eight rats; and in Group 3, 80 µg/kg of cerulein was subcutaneously injected into eight rats. In addition, 30 mg/kg of alpha-tocopherol was intraperitoneally injected into eight rats. RESULTS: The mean Schoenberg score, serum amylase, and lipase and Neutrophil Gelatinase-Associated Lipocalin (NGAL) levels were statistically significantly higher in Group 2 than in Group 1. The mean Schoenberg score and serum amylase and lipase levels were statistically significantly lower in Group 3 than in Group 2. CONCLUSION: In this experimental study rat model of cerulein-induced acute pancreatitis, 30 mg/kg of alpha-tocopherol was injected intraperitoneally to examine its effect on pancreatitis. The improvement was observed in the histopathological examination of pancreatic tissues. We think that alpha-tocopherol may have a therapeutic effect on pancreatic tissue.

Evaluation of the Postmortem Glucose and Glycogen Levels in Hepatic, Renal, Muscle, and Brain Tissues: Is It Possible to Estimate Postmortem Interval Using These Parameters?

The aim of our study was to investigate the postmortem levels of glucose and glycogen in hepatic, renal, muscle, and brain tissues and then examine the changes in those levels that could be useful for estimating postmortem interval. We established an animal model. Seventy female BALB/c albino mice were used in this study. After being sacrificed, the mice were randomly divided into six groups according to time elapsed since death (Group 1: 0 h; Group 2: 12 h; Group 3: 24 h; Group 4: 36 h; Group 5: 48 h; and Group 6: 60 h). Glucose levels were significantly different between groups for all tissues studied. Slope of the change per unit time was higher for the hepatic glucose levels. Based on these results, it is possible to estimate postmortem interval using postmortem glucose levels in hepatic tissue. Tissue-specific assessment may contribute valuable information to postmortem interval studies.

Pentraxin 3 and C-reactive protein as inflammatory markers after a kidney transplant.

OBJECTIVES: There are numerous changes in inflammatory status that occur after a kidney transplant. Pentraxin 3 is a marker of inflammation, but little information is available about pentraxin 3 levels after a kidney transplant. We evaluated the relation between pentraxin 3 and other inflammatory markers including high sensitivity C-reactive protein, interleukin 6, and tumor necrosis factor alpha in kidney transplant recipients. MATERIALS AND METHODS: Adult patients (40 patients; aged, 18-80 y; mean age, 38 ± 10 y) who had a kidney transplant from living-related donors were studied. Patients who had comorbidities associated with chronic inflammation were excluded. Blood samples were obtained before starting immunosuppressive treatment and 2 months after kidney transplant for measurement of pentraxin 3, high sensitivity C-reactive protein, interleukin 6, and tumor necrosis factor α levels. RESULTS: After transplant, mean levels of high sensitivity C-reactive protein and interleukin 6 decreased but levels of pentraxin 3 and tumor necrosis factor alpha did not change. There were significant correlations between interleukin 6 and high sensitivity C-reactive protein before transplant (r = 0.71; P ≤ .0001) and after transplant (r = 0.45; P ≤ .003). There was no correlation between tumor necrosis factor alpha and high sensitivity C-reactive protein before transplant, but there was a significant correlation between tumor necrosis factor alpha and high sensitivity C-reactive protein after transplant (r = 0.36; P ≤ .03). There was no correlation between interleukin 6 and pentraxin 3, tumor necrosis factor alpha and pentraxin 3, or high sensitivity C-reactive protein and pentraxin 3 before or after transplant. CONCLUSIONS: After a kidney transplant, pentraxin 3 may not be useful in determining inflammatory status, and high sensitivity C-reactive protein may be better than pentraxin 3 as a marker of inflammation.

Association between serum cystatin C levels and coronary slow flow.

We assessed the association between serum cystatin C (CysC) levels and coronary slow flow (CSF) phenomenon in patients who underwent coronary angiography. A total of 210 patients (mean age 55.6 ± 10.9 years; 100 females) were included. Patients were divided into 3 groups, (patients with CSF [group 2], with coronary artery disease [CAD; group 3], and without CAD [group 1]). Serum CysC levels in patients with CSF were significantly lower than those with and without CAD (912.5 ± 135.6, 820.4 ± 104.2, and 1343.4 ± 236.6 ng/mL in groups 1, 2, and 3, respectively; P < .001). Serum CysC levels correlated with the number of vessels with CSF and mean corrected thrombolysis in myocardial infarction frame count (Spearman correlation coefficient [r s] = .192, P < .001 and r s = .261, P < .001 respectively). In conclusion, patients with CSF have lower CysC levels; this could be a useful biomarker of CSF involvement in patients who undergo diagnostic coronary angiography.