Publications by authors named "Tsvetelina Velikova"

40 Publications

Biomarkers in Irritable Bowel Syndrome: biological rationale and diagnostic value.

Dig Dis 2021 Mar 22. Epub 2021 Mar 22.

Patients with irritable bowel syndrome (IBS) usually suffer from non-specific and overlapping signs that hamper the diagnostic process. In line with this, biomarkers specific for IBS could be of great benefit for diagnosing and managing patients. In IBS, the need is for apparent distinguishing features linked to the disease that improve diagnosis, differentiate from other organic diseases, and discriminate between IBS subtypes. Some biomarkers are associated with a possible pathophysiologic mechanism of IBS; others are used for differentiating IBS from non-IBS patients. Implementation of IBS biomarkers in everyday clinical practice is critical for early diagnosis and treatment. However, our knowledge about their efficient use is still scarce. This review discusses the biomarkers implemented for IBS diagnosis and management, such as blood (serum), fecal, immunological, related to the microbiome, microRNAs, and some promising novel biomarkers associated with imaging and psychological features of the disease. We focus on the most common, studied, and validated biomarkers and their biological rationale, diagnostic, and clinical value.
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http://dx.doi.org/10.1159/000516027DOI Listing
March 2021

Neopterin in the Evolution from Obesity to Prediabetes and Newly Diagnosed Type 2 Diabetes.

Metab Syndr Relat Disord 2021 May 16;19(4):249-255. Epub 2021 Feb 16.

Clinic of Endocrinology, Department of Internal Medicine, Medical University of Sofia, University Hospital "Alexandrovska," Sofia, Bulgaria.

Neopterin, marker of cellular immunity and oxidative stress, is mainly produced by activated macrophages. It could play a crucial role in the development of insulin resistance (IR) and type 2 diabetes (T2D). The aim of this study was to investigate the circulating levels of neopterin in different stages of glucose dysregulation from obesity through prediabetes to newly diagnosed diabetes. Neopterin levels were determined using a commercially available human enzyme-linked immunosorbent assay kit. The homeostasis model assessment of IR was used as an index to assess IR. The sample consisted of 163 subjects with mean age 52.5 ± 11.3 years, divided in three age- and body mass index (BMI)-matched groups-obesity, prediabetes, and diabetes. The control group consisted of 42 healthy individuals. Neopterin levels were significantly higher in patients with obesity and/or prediabetes and newly diagnosed diabetes than those in the control group, respectively (4.14 ± 2.51; 4.04 ± 2.80 and 2.17 ± 1.93 vs. 0.87 ± 0.84;  < 0.05). Correlation analysis showed that the level of neopterin positively correlated with BMI, waist, waist-to-stature ratio, waist-to-hip ratio, fasting glucose, and triglycerides. Receiver operating characteristic analysis established neopterin suitable for distinguishing subjects with obesity [area under the curve (AUC) = 0.83;  < 0.001] and carbohydrate disturbances (AUC = 0.59;  < 0.05) from those without these conditions. Neopterin ≥0.47 ng/mL have an odds ratio (OR) of 2.71 for development of dysglycemia, whereas threshold value of neopterin ≥0.56 ng/mL shows an OR of 5.94 for development of obesity. The levels of neopterin were increased in patients with obesity and carbohydrate disturbances. Further studies will elucidate the role of the biomarker in development of T2D and its complications.
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http://dx.doi.org/10.1089/met.2020.0144DOI Listing
May 2021

Antibiotic-Related Changes in Microbiome: The Hidden Villain behind Colorectal Carcinoma Immunotherapy Failure.

Int J Mol Sci 2021 Feb 10;22(4). Epub 2021 Feb 10.

Department of Gastroenterology, University Hospital Lozenetz, Sofia University St. Kliment Ohridski, Kozyak 1 Str., 1407 Sofia, Bulgaria.

The interplay between drugs and microbiota is critical for successful treatment. An accumulating amount of evidence has identified the significant impact of intestinal microbiota composition on cancer treatment response, particularly immunotherapy. The possible molecular pathways of the interaction between immune checkpoint inhibitors (ICIs) and the microbiome can be used to reverse immunotherapy tolerance in cancer by using various kinds of interventions on the intestinal bacteria. This paper aimed to review the data available on how the antibiotic-related changes in human microbiota during colorectal cancer (CRC) treatment can affect and determine ICI treatment outcomes. We also covered the data that support the potential intimate mechanisms of both local and systemic immune responses induced by changes in the intestinal microbiota. However, further better-powered studies are needed to thoroughly assess the clinical significance of antibiotic-induced alteration of the gut microbiota and its impact on CRC treatment by direct observations of patients receiving antibiotic treatment.
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http://dx.doi.org/10.3390/ijms22041754DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7916407PMC
February 2021

SARS-CoV-2 vaccines and autoimmune diseases amidst the COVID-19 crisis.

Rheumatol Int 2021 03 30;41(3):509-518. Epub 2021 Jan 30.

First Department of Internal Medicine, Faculty of Medicine, Medical University-Varna, 55 Marin Drinov Str., Varna, 9002, Bulgaria.

Coronavirus disease 2019 (COVID-19) pandemic has become challenging even for the most durable healthcare systems. It seems that vaccination, one of the most effective public-health interventions, presents a ray of hope to end the pandemic by achieving herd immunity. In this review, we aimed to cover aspects of the current knowledge of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines and vaccine candidates in the light of autoimmune inflammatory diseases (AIIDs) and to analyze their potential in terms of safety and effectiveness in patients with AIIDs. Therefore, a focused narrative review was carried out to predict the possible implications of different types of SARS-CoV-2 vaccines which confer distinct immune mechanisms to establish immune response and protection against COVID-19: whole virus (inactivated or weakened), viral vector (replicating and non-replicating), nucleic acid (RNA, DNA), and protein-based (protein subunit, virus-like particle). Still, there is uncertainty among patients with AIIDs and clinicians about the effectiveness and safety of the new vaccines. There are a variety of approaches towards building a protective immunity against SARS-CoV-2. Only high-quality clinical trials would clarify the underlying immunological mechanisms of the newly implemented vaccines/adjuvants in patients living with AIIDs.
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http://dx.doi.org/10.1007/s00296-021-04792-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7846902PMC
March 2021

Current concept for tertiary lymphoid structures in urothelial carcinoma of the bladder: a literature review and our experience.

Am J Clin Exp Immunol 2020 15;9(5):64-72. Epub 2020 Dec 15.

Department of Clinical Immunology, Medical University of Sofia, Bulgaria/University Hospital Lozenetz Sofia, Bulgaria.

Bladder carcinoma (BC) is one of the most common malignancies of the urinary system in developed countries, with a high number of recurrences. The secondary lymphoid organs (SLO) are crucial for initiating the adaptive immune response. They are developed as a part of a genetically preprogrammed process during embryogenesis. However, SLO's organogenesis can be reduplicated de novo in other tissues by a process termed lymphoid neo-genesis, giving rise to tertiary lymphoid structures (TLS). These well-organized lymphoid structures in cancer are essential modulators of cancer immunologic response, and the histological examination of TLS gave a new strategy for cancer immunotherapy. This review explores the biological and histological characteristics of TLS in muscle non-invasive and invasive BC.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7811926PMC
December 2020

Concentrations of Interleukin-1ß in Gingival Crevicular Fluid and Saliva - a Potential Diagnostic Biomarker of Periodontal Diseases.

Folia Med (Plovdiv) 2020 Dec;62(4):825-830

Medical University of Sofia, Sofia, Bulgaria.

Introduction: Bacterial challenge in periodontal diseases activates both local and systemic immune responses of a macroorganism by increasing multiple proinflammatory factors that can be discovered in gingival crevicular fluid (GCF) and in saliva. We tested the hypothesis that IL-1β concentration in GCF and saliva correlates with periodontal health and diseases. Materials and methods: The study included 62 people (mean age 36±14 yrs), divided into three groups - patients with periodontitis (24 people), patients with gingivitis (19 people) and periodontally healthy people (19 people). Saliva and GCF samples were taken from all participants and the levels of IL-1β in all samples were determined by ELISA.

Results: IL-1β concentrations in GCF of healthy individuals were significantly lower than the IL-1β concentration in GCF of patients with gingivitis (p=0.009) and with periodontitis (p.
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http://dx.doi.org/10.3897/folmed.62.e49872DOI Listing
December 2020

What could microRNA expression tell us more about colorectal serrated pathway carcinogenesis?

World J Gastroenterol 2020 Nov;26(42):6556-6571

Medical Faculty, Sofia University St. Kliment Ohridski, Sofia 1407, Bulgaria.

In the last two decades, the vision of a unique carcinogenesis model for colorectal carcinoma (CRC) has completely changed. In addition to the adenoma to carcinoma transition, colorectal carcinogenesis can also occur the serrated pathway. Small non-coding RNA, known as microRNAs (miRNAs), were also shown to be involved in progression towards malignancy. Furthermore, increased expression of certain miRNAs in premalignant sessile serrated lesions (SSLs) was found, emphasizing their role in the serrated pathway progression towards colon cancer. Since miRNAs function as post-transcriptional gene regulators, they have enormous potential to be used as useful biomarkers for CRC and screening in patients with SSLs particularly. In this review, we have summarized the most relevant information about the specific role of miRNAs and their relevant signaling pathways among different serrated lesions and polyps as well as in serrated adenocarcinoma. Additional focus is put on the correlation between gut immunity and miRNA expression in the serrated pathway, which remains unstudied.
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http://dx.doi.org/10.3748/wjg.v26.i42.6556DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7673963PMC
November 2020

Serum levels of matrix metalloproteinase-3 as a prognostic marker for progression of cartilage injury in patients with knee osteoarthritis.

Acta Reumatol Port 2020 Jul-Sep;45(3):207-213

University Hospital "St. Ivan Rilski", Medical Faculty, Medical University - Sofia.

Objective: To evaluate serum matrix metalloproteinase (MMP)-3 levels as a prognostic marker for the progression of cartilage damage in patients with knee osteoarthritis (KOA).

Methods: Fifty-six patients, aged 40 to 80 years (62.59 ± 10.11 years) who met the ACR criteria for KOA, were included in a one-year observational prospective clinical study. Complete baseline and follow-up data were collected from 50 out of 56 patients. X-ray and magnetic-resonance images were carried out at baseline and after 12 months. They were evaluated according to the Kellgren-Lawrence and Whole-Organ magnetic Resonance iMaging Score (WORMS) semi-quantitative scales, respectively. Progression of cartilage damage in the medial tibiofemoral compartment was registered at the end of the follow-up using the change in WORMS. Serum levels of MMP-3 were measured during the baseline visit, using enzyme-linked immunosorbent assay.

Results: Significantly higher values of baseline MMP-3 levels were observed in patients with a registered progression of cartilage injury in the medial tibiofemoral compartment of the knee compared with patients with no progression (p = 0.005). Binary logistic regression analysis showed that levels of serum MMP-3 (ng/ml) were an independent predictor of subsequent progression of cartilage injury in the medial tibiofemoral compartment of the index knee (assessed by MRI) (OR = 1.042, CI 95%: 1.002-1.084). Receiver operating characteristic analysis was performed to delineate progressors from non-progressors.

Conclusion: Serum MMP-3 levels may serve as a potential prognostic biomarker for cartilage injury in patients with KOA.
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November 2020

Increased PD-L1 expression in high-grade bladder cancer with squamous cell differentiation in Bulgarian and French patients' samples.

Ann Diagn Pathol 2020 Dec 8;49:151640. Epub 2020 Oct 8.

Department of General and Clinical Pathology, Medical University/University Hospital "St.George", Plovdiv, Bulgaria.

Urothelial carcinomas (UC) of the bladder are biologically and clinically heterogeneous and the most common malignancy of the urinary tract in developed countries worldwide, where several checkpoint targets as programmed death ligand-1 (PD-L1) and programmed cell death protein (PD-1) have received the most attention in the treatment of bladder cancer. However, the clinicopathological impact of this biomarker has not yet been established enough.

Objective: To evaluate programmed death ligand-1 (PD-L1) expression in UCs of the bladder in Bulgarian and French patients' samples.

Materials And Methods: Urothelial bladder carcinomas cases from 2016-2020 were retrospectively were analyzed. The cohort included 105 cases: 42 (40%) low grade and 63 (60%) high grade. Immunohistochemical (IHC) staining for PD-L1 expression was performed using an anti-PD-L1 primary antibody clone 22C3pharmDx only to 73/105 cases.

Results: Approximately 21/73 cases (28.8%) of urothelial bladder carcinomas demonstrated positive PD-L1 expression, and in 52/73 cases (71.2%) were negative. Positive PD-L1 expression was associated with high grade and high pathologic stage (p < 0.001). We found that PD-L1 was expressed in a significant percentage in UC with squamous differentiation (40%), followed by classic UC (30%). An association between histological grading systems of bladder UC (WHO1973 and WHO 2016) and the TNM-staging system, estimated by Pearson correlation coefficients (r = 0.590 and r = 0.583, respectively, p < 0.001) was observed.

Conclusions: We found that PD-L1 expression is increased in patients with muscle-invasive UC, and PD-L1 might be a new biomarker that correlates with the pathological stage of urothelial bladder cancer and might predict recurrence-free survival.
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http://dx.doi.org/10.1016/j.anndiagpath.2020.151640DOI Listing
December 2020

Immunological aspects of COVID-19: What do we know?

World J Biol Chem 2020 Sep;11(2):14-29

Department of Epidemiology and Disaster Medicine, Medical University, Plovdiv, University Hospital "St George", Plovdiv 4002, Bulgaria.

The newly emerged coronavirus (severe acute respiratory syndrome coronavirus 2 SARS-CoV-2) and the disease that it causes coronavirus disease 2019 (COVID-19) have changed the world we know. Yet, the origin and evolution of SARS-CoV-2 remain mostly vague. Many virulence factors and immune mechanisms contribute to the deteriorating effects on the organism during SARS-CoV-2 infection. Both humoral and cellular immune responses are involved in the pathophysiology of the disease, where the principal and effective immune response towards viral infection is the cell-mediated immunity. The clinical picture of COVID-19, which includes immune memory and reinfection, remains unclear and unpredictable. However, many hopes are put in developing an effective vaccine against the virus, and different therapeutic options have been implemented to find effective, even though not specific, treatment to the disease. We can assume that the interaction between the SARS-CoV-2 virus and the individual's immune system determines the onset and development of the disease significantly.
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http://dx.doi.org/10.4331/wjbc.v11.i2.14DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7520644PMC
September 2020

ABO-nonidentical liver transplantation from a deceased donor and clinical outcomes following antibody rebound: A case report.

World J Transplant 2020 May;10(5):138-146

Department of Clinical Immunology, University Hospital Lozenetz, Sofia 1407, Bulgaria.

Background: Although ABO-nonidentical and ABO-incompatible liver transplantation (LT) are other options for end-stage liver disease treatment, the development of antibodies against blood group antigens (anti-A/B antibodies) is still a challenge in managing and follow-up of the recipients.

Case Summary: A 56-year-old male with end-stage liver disease with rapid deterioration and poor prognosis was considered to receive a deceased ABO-nonidentical liver graft. All required tests were performed according to our pre-LT diagnostic protocol. The orthotopic LT procedure involving O+ donor and A1B+ recipient was performed. Our treatment strategy to overcome the antibody-mediated rejection included a systemic triple immunosuppressive regimen: methylprednisolone, mycophenolate mofetil, and tacrolimus. The immunological desensitization consisted of the chimeric anti-CD20 monoclonal antibody rituximab and intravenous immunoglobulins. The patient was also on antibiotic treatment with amoxicillin/clavulanate, cefotaxime, and metronidazole. On the 10 postoperative day, high titers of IgG anti-A and anti-B antibodies were found in the patient's plasma. We performed a liver biopsy, which revealed histological evidence of antibody-mediated rejection, but the rejection was excluded according to the Banff classification. The therapy was continued until the titer decreased significantly on the 18 postoperative day. Despite the antibiotic, antifungal, and antiviral treatment, the patient deteriorated and developed septic shock with anuria and pancytopenia. The conservative treatment was unsuccessful, which lead to the patient's fatal outcome on the 42 postoperative day.

Conclusion: We present a patient who underwent ABO-nonidentical LT from a deceased donor. Even though we implemented the latest technological advancements and therapeutic approaches in the management of the patient and the initial results were promising, due to severe infectious complications, the outcome was fatal.
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http://dx.doi.org/10.5500/wjt.v10.i5.138DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7428789PMC
May 2020

Lumican in Obese Patients with Nonalcoholic Fatty Liver Disease With or Without Prediabetes.

Metab Syndr Relat Disord 2020 11 11;18(9):443-448. Epub 2020 Aug 11.

Department of Internal Medicine, Clinic of Endocrinology and Metabolic Diseases, University Hospital "Alexandrovska," Medical University, Sofia, Bulgaria.

Lumican is a small leucine-rich proteoglycan that regulates the assembly of collagen fibers in the extracellular matrix of different tissues. Excess collagen production in the liver is key in the pathogenesis of nonalcoholic fatty liver disease (NAFLD) and might contribute to the risk of type 2 diabetes mellitus and cardiovascular diseases. The aim of this study was to evaluate the relationship between serum lumican and prediabetes, and other biochemical and clinical parameters in obese subjects with NAFLD. The study group included 79 subjects with obesity and NAFLD of which 41 had normal carbohydrate tolerance and 38 had prediabetes. Serum lumican was measured by means of enzyme-linked immunosorbent assay. Higher lumican serum levels were found in patients with prediabetes compared with those with normal carbohydrate tolerance (0.117 ± 0.074 vs. 0.080 ± 0.048 ng/mL,  = 0.010) as well as in subjects with metabolic syndrome (MetS) versus those without MetS (0.113 ± 0.071 vs. 0.079 ± 0.048 ng/mL,  = 0.034). There was also a modest positive association between lumican levels and fasting glucose ( = 0.228,  < 0.05). Lumican levels ≥0.07 ng/mL determine a 3.9-fold higher risk of prediabetes (odds ratio: 3.945, 95% confidence interval: 1.518-10.254,  = 0.005). Lumican levels were higher in obese subjects with NAFLD with prediabetes and MetS. Lumican bears an increased risk for prediabetes in the study population.
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http://dx.doi.org/10.1089/met.2020.0001DOI Listing
November 2020

Relationship between circulating netrin-1 levels, obesity, prediabetes and newly diagnosed type 2 diabetes.

Arch Physiol Biochem 2020 Jul 11:1-6. Epub 2020 Jul 11.

Department of Internal Medicine, Medical University, Sofia, Bulgaria.

Background: Netrin-1 is presumed to have regenerative, angiogenic and anti-inflammatory properties, thus it could play a substantial role in the development of insulin resistance and T2DM.

Objective: The aim of this study was to evaluate the relationship between serum netrin-1 levels and carbohydrate disturbances in patients with obesity.

Methods: Sample size consisted of 163 patients, divided into four groups: obesity without carbohydrate disturbances prediabetes and diabetes and healthy controls Netrin-1 level was determined using ELISA method.

Results: Circulating serum Netrin-1 was significantly lower in patients only with obesity, as well as with those with prediabetes and diabetes in comparison to the control group. Correlation analysis revealed that netrin-1 correlates negatively with BMI, waist, WSR, LDL and positive with sudomotor function. Netrin-1 ≤ 0.17 ng/ml has about 3 fold higher risk for carbohydrate disturbances (OR 3.06, 95% CI 1.48-6.34,  = .003).

Conclusion: Netrin-1 is associated with an increased risk for glycaemic disorders in patients with obesity.
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http://dx.doi.org/10.1080/13813455.2020.1780453DOI Listing
July 2020

Identification of Specific IgE Antibodies and Asthma Control Interaction and Association Using Cluster Analysis in a Bulgarian Asthmatic Children Cohort.

Antibodies (Basel) 2020 Jul 6;9(3). Epub 2020 Jul 6.

Medical University, Pediatric clinic, UMHAT Alexandrovska, 1 Georgi Sofiyski blvd., 1431 Sofia, Bulgaria.

(1) Background: Asthma is a complex heterogeneous disease that likely comprises several distinct disease phenotypes, where the clustering approach has been used to classify the heterogeneous asthma population into distinct phenotypes; (2) Methods: For a period of 1 year, we evaluated medical history data of 71 children with asthma aged 3 to 17 years, performing pulmonary function tests, drew blood for IgE antibodies against inhalation and food allergies detection, and Asthma Control Questionnaire (ACQ); (3) Results: Five distinct phenotypes were determined. Cluster 1 (n = 10): (non-atopic) the lowest IgE level, very low ACQ, and median age of diagnosis. Cluster 2 (n = 28): (mixed) the highest Body mass index (BMI) with the latest age of diagnosis and high ACQ and bronchodilator response (BDR) levels and median and IgE levels. Cluster 3 (n = 19) (atopic) early diagnosis, highest BDR, highest ACQ score, highest total, and high specific IgE levels among the clusters. Cluster 4 (n = 9): (atopic) the highest specific IgE result, relatively high BMI, and IgE with median ACQ score among clusters. Cluster 5 (n = 5): (non-atopic) the earliest age for diagnosis, with the lowest BMI, the lowest ACQ score, and specific IgE levels, with high BDR and the median level of IgE among clusters; (4) Conclusions: We identified asthma phenotypes in Bulgarian children according to IgE levels, ACQ score, BDR, and age of diagnosis.
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http://dx.doi.org/10.3390/antib9030031DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7551616PMC
July 2020

Lactobacilli Supplemented with Larch Arabinogalactan and Colostrum Stimulates an Immune Response towards Peripheral NK Activation and Gut Tolerance.

Nutrients 2020 Jun 7;12(6). Epub 2020 Jun 7.

Laboratory of Clinical Immunology, University Hospital "St. Ivan Rilski", Medical University of Sofia, 15 "Acad. Iv. Evst. Geshov" Blvd, 1431 Sofia, Bulgaria.

Probiotics possibly affect local and systemic immune reactions and maintain the intestinal immune homeostasis in healthy individuals and patients with diseases such as irritable bowel syndrome (IBS). In this single-center, blinded trial, we enrolled 40 individuals (20 patients with IBS and 20 healthy individuals) whose blood and fecal samples were collected before and after a 21-day administration of a product comprising spp., larch arabinogalactan, and colostrum. The percentage of HLA-DR+ natural killer (NK) cells was higher in healthy individuals ( = 0.03) than in patients with IBS after product supplementation. In the fecal samples of patients with IBS, we observed a decline in IL-6, IFN-γ, TNF-α, and secretory IgA levels and, simultaneously, an increase in IL-10 and IL-17A levels after supplementation, although non-significant, whereas, in healthy individuals, we observed a significant decline in IL-6 and IFN-γ levels after supplementation ( < 0.001). Nevertheless, we observed a clinical improvement of symptoms in 65-75% of patients with IBS and the complete resolution of the initial symptoms in five of the 20 patients. We also observed a possible prophylactic effect by the inducing system antiviral impact accompanied by a trend for local immune tolerance in the gut in healthy individuals, where it is the desirable state.
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http://dx.doi.org/10.3390/nu12061706DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7352398PMC
June 2020

Impact of testosterone treatment on circulating irisin in men with late-onset hypogonadism and metabolic syndrome.

Aging Male 2020 Dec 26;23(5):1381-1387. Epub 2020 May 26.

Department of Internal Diseases, Medical University of Sofia, Sofia, Bulgaria.

Objectives: The beneficial effects of testosterone replacement therapy (TRT) in men with late-onset hypogonadism (LOH) on the body composition and metabolic outcomes are well-established. A potential explanation might lie in the hormones, secreted from skeletal muscles, named "myokines". The aim of this study was to evaluate the effects of TRT on the levels of serum irisin in subjects with LOH.

Study Design: A total 40 men with metabolic syndrome (MS) and LOH (measured serum testosterone concentration < 12 nmol/l). TRT with Testosterone Undecanoate (Nebido™) was performed at baseline and at week 6. Irisin serum concentration was determined at baseline and at week 18 by means of ELISA.

Results: Circulating irisin was positively associated with serum testosterone (r = 0.283,  < 0.05). TRT has led to a statistically significant rise in circulating serum irisin levels (7.12 ± 0.76 mcg/ml versus 7.76 ± 0.75 mcg/ml; paired-samples t-test  < 0.001). ROC-analyses determined irisin to be predictive of treatment response (AUC = 0.741,  = 0.014).

Conclusions: Irisin is positively associated with serum testosterone in a population of men with MS and LOH. TRT in these subjects has led to a significant improvement in associated clinical symptoms as well as to a significant rise in serum irisin levels.
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http://dx.doi.org/10.1080/13685538.2020.1770721DOI Listing
December 2020

Interleukin-6 compared to the other Th17/Treg related cytokines in inflammatory bowel disease and colorectal cancer.

World J Gastroenterol 2020 Apr;26(16):1912-1925

Department of Molecular Biology, Immunology and Medical Genetics, Medical Faculty, Trakia University, Stara Zagora 6000, Bulgaria.

Background: The connection between inflammatory bowel disease (IBD) and colorectal cancer (CRC) is well-established, as persistent intestinal inflammation plays a substantial role in both disorders. Cytokines may further influence the inflammation and the carcinogenesis process.

Aim: To compare cytokine patterns of active IBD patients with early and advanced CRC.

Methods: Choosing a panel of cytokines crucial for Th17/Treg differentiation and behavior, in colon specimens, as mRNA biomarkers, and their serum protein levels.

Results: We found a significant difference between higher gene expression of , , , and , and approximately equal level of in CRC patients in comparison with IBD patients. After stratification of CRC patients, we found a significant difference in , , , and mRNA in early cases compared to IBD, and alone in advanced CRC. The protein levels of the cytokines were significantly higher in CRC patients compared to IBD patients.

Conclusion: Our findings showed that upregulation is essential for both IBD and CRC development until the upregulation of other Th17/Treg related genes (, and transcription factor ) is a crucial primarily for CRC development. The significantly upregulated could be a potential drug target for IBD and prevention of CRC development as well.
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http://dx.doi.org/10.3748/wjg.v26.i16.1912DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7201144PMC
April 2020

Challenges and updates in the management of eosinophilic esophagitis.

Prz Gastroenterol 2020 12;15(1):27-33. Epub 2019 Apr 12.

Department of Clinical Immunology, University Hospital Lozenetz, Sofia, Bulgaria.

Eosinophilic esophagitis (EE) is a chronic T helper cell-2 mediated inflammatory condition and is considered as a primary cause of dysphagia and foregut symptoms. There are many challenges regarding the treatment options of EE. Different therapeutic approaches are best at meeting different endpoints. There are unresolved questions such as whether the goal for treatment should be to control esophageal eosinophilia and inflammation or to achieve symptomatic improvement. Still, proton pump inhibitors are used in differential diagnosis, along with their anti-inflammatory and anti-acid properties. Conducted trials continue to recommend the use of topical corticosteroids and empiric food elimination diets as first-line therapeutic modalities. The growing knowledge on the pathogenesis of EE has allowed further progression of encouraging targeted biologic therapies. However, the guidelines for EE management should be updated accordingly in the coming years, including fast evolving data on non-invasive diagnostic strategies, new treatment modalities, and the long-term prognosis of the disease.
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http://dx.doi.org/10.5114/pg.2019.84476DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7089863PMC
April 2019

Chemical Metabolism of Xenobiotics by Gut Microbiota.

Curr Drug Metab 2020 ;21(4):260-269

Clinical Immunology, Lozenetz University Hospital, Sofia, Bulgaria.

Among the gut microbiota's newly explored roles in human biology is the ability to modify the chemical structures of foreign compounds (xenobiotics). A growing body of evidence has now provided sufficient acumen on the role of the gut microbiota on xenobiotic metabolism, which could have an intense impact on the therapy for various diseases in the future. Gut microbial xenobiotic metabolites have altered bioavailability, bioactivity and toxicity and can intervene with the actions of human xenobiotic-metabolizing enzymes to affect the destiny of other ingested molecules. These modifications are diverse and could lead to physiologically important consequences. In the current manuscript we aim to review the data currently available on how the gut microbiota directly modifies drugs, dietary compounds, chemicals, pollutants, pesticides and herbal supplements.
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http://dx.doi.org/10.2174/1389200221666200303113830DOI Listing
May 2021

Retraction notice to "Alterations in cytokine gene expression profile in colon mucosa of inflammatory bowel disease patients on different therapeutic regimens" [Cytokine 92 (2017) 12-19].

Cytokine 2020 05 20;129:155018. Epub 2020 Feb 20.

Department of Molecular Biology, Immunology and Medical Genetics, Medical Faculty, Trakia University, Armeiska 11 Str., Stara Zagora 6000, Bulgaria.

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http://dx.doi.org/10.1016/j.cyto.2020.155018DOI Listing
May 2020

The association between retinol-binding protein 4 and prediabetes in obese patients with nonalcoholic fatty liver disease.

Arch Physiol Biochem 2019 Oct 7:1-6. Epub 2019 Oct 7.

Department of Internal Medicine, Clinic of Gastroenterology, University Hospital "St. Ivan Rilski", Medical University - Sofia , Sofia , Bulgaria.

Retinol-binding protein 4 RBP4) is associated with visceral fat and insulin resistance (IR) in obesity, type 2 diabetes mellitus (T2DM) and nonalcoholic fatty liver disease (NAFLD), but some of these data remain controversial. This study evaluated the relationship between serum RBP4 levels and prediabetes in obese patients with NAFLD. A total of 79 obese NAFLD patients without ( = 41) and with prediabetes ( = 38) were included. Serum RBP4 was measured using ELISA method. Higher RBP4 serum levels were observed in patients with prediabetes, metabolic syndrome (MetS), or dyslipidaemia. There was correlation between RBP4 levels and visceral adiposity index (VAI), glucose, insulin, HOMA-IR, and Quicki index. RBP4 ≥ 61 mcg/ml have about 3.5-fold higher risk of prediabetes (OR 3.544, 95% CI 1.385-9.072, =.008), and RBP4 ≥ 55 mcg/ml increased the risk for MetS approximately 3.1 times. RBP4 is associated with increased risk for prediabetes and MetS in obese patients with NAFLD.
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http://dx.doi.org/10.1080/13813455.2019.1673429DOI Listing
October 2019

The Dynamic Changes in Soluble CD30 and Regulatory T Cells Before and After Solid Organ Transplantations: A Pilot Study.

Monoclon Antib Immunodiagn Immunother 2019 Aug 30;38(4):137-144. Epub 2019 Jul 30.

7Department of Surgery, University Hospital "Lozenetz," Sofia University "St. Kliment Ohridski," Sofia, Bulgaria.

Among multiple parameters, applied in the immunologic monitoring of transplantation, the levels of serum soluble CD30 (sCD30) and peripheral regulatory T cells (Tregs) are very promising. These are relatively new biomarkers, considered to reflect immune activation and tolerance in solid organ transplantation. Results are shown here from a preliminary study on the relevance of sCD30 and Tregs in the monitoring of the early post-transplantation period. Sixteen patients with chronic liver or kidney disease were examined. Nine of them were further selected for transplantation. Follow-up of sCD30 and Tregs was carried out during the first month after transplantation. Until day 30 (D30) after transplantation, a progressive decrease in sCD30 levels was observed in all patients. Conversely, the dynamic of Tregs was dependent on the transplanted organ: in liver recipients, an increase of Tregs was detected at day 7 (D7) followed by a gradual decrease until D30, whereas in kidney recipients, a sustained downward trend starting on D7 was observed. In liver recipients, the increase in Tregs preceded albumin normalization, whereas in kidney recipients, sCD30 was found to have predictive significance for the creatinine levels. Our results demonstrated that peripheral blood sCD30 and Tregs are valuable parameters in the immunologic monitoring of transplanted patients.
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http://dx.doi.org/10.1089/mab.2019.0010DOI Listing
August 2019

Serum levels of IL-5, IL-6, IL-8, IL-13 and IL-17A in pre-defined groups of adult patients with moderate and severe bronchial asthma.

Respir Med 2019 Jul - Aug;154:144-154. Epub 2019 Jun 26.

Clinical Immunology, University Hospital Lozenetz, Bulgaria. Electronic address:

Background: Bronchial asthma (BA) is a complex disease characterised by persistent inflammation. Exhaled nitric oxide (FeNO) and blood eosinophil count (b-Eos) are biomarkers for type 2 endotype of BA.

Objective: To analyse a panel of serum interleukins and total IgE in predefined by FeNO and b-Eos groups of moderate and severe BA patients.

Methods: Serum levels of IL-5, IL-6, IL-8, IL-13 and IL-17A (ELISA) were measured in 30 healthy controls (HC) and 80 adult BA patients. BA patients were split into 4 groups. Group 1:Low FeNO/Low b-Eos (n = 23; 28.8%); Group 2:Low FeNO/High b-Eos (n = 17; 21.3%); Group 3:High FeNO/Low b-Eos (n = 15; 18.8%); Group 4:High FeNO/High b-Eos (n = 25; 31.3%).

Results: All interleukins and total IgE were significantly higher in patients with BA as compared with HC. IL-5 levels were highest in Group 2 (p < 0.05). IL-6, IL-13 and IL-17A levels were elevated in Groups 2, 3 and 4 as compared with HC (p < 0.05). Higher IL-8 levels were associated with a pattern of current smokers. Highest IL-17A levels were found in type 2 high groups with frequent exacerbations, mostly uncontrolled and severe BA. We have found a distinct pattern for each group based on demographic, clinical, functional, immunological and inflammatory characteristics.

Conclusion: FeNO and b-Eos are useful in the identification of severe type 2 BA subgroups with frequent exacerbations. IL-5, IL-6, IL-13 and IL-17A are involved in the persistent type 2 immune response in moderate and severe BA. We have identified a pattern of refractory, severe type 2/IL-17A high BA in the real clinical practice.
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http://dx.doi.org/10.1016/j.rmed.2019.06.024DOI Listing
July 2020

Trefoil Factor 3 is Highly Predictive of Complete Mucosal Healing Independently and in Combination with C-Reactive Protein in Patients with Ulcerative Colitis.

J Gastrointestin Liver Dis 2019 Jun 1;28:169-174. Epub 2019 Jun 1.

Clinic of Gastroenterology, Tsaritsa Yoanna University Hospital, Medical University of Sofia, Sofia, Bulgaria.

Aim: In the current study we aimed to evaluate the role of trefoil factor 3 (TFF3) as a marker for complete mucosal healing (MH) in patients with ulcerative colitis (UC).

Methods: We enrolled 116 consecutive UC patients. Trefoil factor 3 levels were measured by ELISA and were compared to the clinical activity, assessed by Lichtiger Index, fecal calprotectin (FCP) and C-reactive protein (CRP) levels. Colonoscopy was performed in all the patients and the findings were graded according to Mayo endoscopic score (EMS) and UC endoscopic index of severity (UCEIS).

Results: Trefoil factor 3 levels were significantly correlated with Lichtiger Index (r=0.736, p<0.001), EMS (r=0.811, p<0.001), UCEIS (r=0.820, p<0.001), FCP (r=0.696, p<0.001) and CRP (r=0.405, p<0.001). The TFF3 cut-off level of 6.74 ng/ml indicated complete MH (EMS=0; UCEIS=0) with a sensitivity and specificity of 0.879 and 0.869, respectively (area under the curve, AUC 0.927; 95% confidence interval, 0.877-0.976). The DeLong's test revealed no significant difference between the AUC of TFF3+CRP and the AUC of FCP (Z=1.717, p=0.086), AUC of TFF3+FCP (Z=1.908, p=0.056), and AUC of TFF3+CRP+FCPP (Z=1.915, p=0.056). However, the AUC of TFF3+CRP showed significant difference compared with the AUC of TFF3 (Z=2.210, p=0.027) and the AUC of CRP (Z=3.145, p=0.002) for predicting complete MH.

Conclusions: Trefoil factor 3 levels correlated significantly with clinical activity, endoscopic indices, CRP and FCP in our patients. TFF3 is a highly predictive marker of complete MH independently and in combination with CRP in patients with UC.
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http://dx.doi.org/10.15403/jgld-177DOI Listing
June 2019

Celiac-Related Autoantibodies and IL-17A in Bulgarian Patients with Dermatitis Herpetiformis: A Cross-Sectional Study.

Medicina (Kaunas) 2019 May 15;55(5). Epub 2019 May 15.

Department of Dermatology, Faculty of Medicine, Medical University-Sofia, 1431 Sofia, Bulgaria.

: Dermatitis herpetiformis (DH) is a blistering dermatosis, which shares common immunologic features with celiac disease (CD). The aim of the present study was to explore the performance of a panel of CD-related antibodies and IL-17A in Bulgarian patients with DH. Serum samples from 26 DH patients at mean age 53 ± 15 years and 20 healthy controls were assessed for anti-tissue transglutaminase (anti-tTG), anti-deamidated gliadin peptides (anti-DGP), anti-actin antibodies (AAA), and IL-17A by enzyme linked immuno-sorbent assay (ELISA), as well as anti-tTG, anti-gliadin (AGA), and anti-Saccharomyces cerevisiae antibodies (ASCA) using immunoblot. The average serum levels of anti-tTG, anti-DGP, AGA, AAA, and the cytokine IL-17A were at significantly higher levels in patients with DH compared to the average levels in healthy persons which stayed below the cut-off value ( < 0.05). Anti-DGP and anti-tTG antibodies showed the highest diagnostic sensitivity and specificity, as well as acceptable positive and negative predictive value. None of the healthy individuals was found positive for the tested antibodies, as well as for ASCA within the DH group. All tests showed good to excellent correlations (r = 0.5 ÷ 0.9, < 0.01). Although the diagnosis of DH relies on skin biopsy for histology and DIF, serologic testing of a panel of celiac-related antibodies could be employed with advantages in the diagnosing process of DH patients. Furthermore, DH patients who are positive for the investigated serologic parameters could have routine monitoring for gastrointestinal complications typical for the gluten-sensitive enteropathy.
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http://dx.doi.org/10.3390/medicina55050136DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6572648PMC
May 2019

Peroxiredoxin 4 levels in patients with PCOS and/or obesity.

J Gynecol Obstet Hum Reprod 2019 Nov 11;48(9):739-743. Epub 2019 Apr 11.

Medical University-Sofia, Bulgaria Department of Internal Medicine, Clinic of Endocrinology, University Hospital "Alexandrovska", Bulgaria.

Background: Peroxiredoxin 4 is a part of endogen antioxidant system and its levels are elevated in increased oxidative stress conditions. It is found to be positively associated with cardiovascular risk. The aim of the study was to investigate peroxiredoxin 4 levels in women with polycystic ovarian syndrome (PCOS) and/or obesity.

Methods: In this cros-sesctional study were included 80 patients. Anthropometric measurements and biochemical tests, including peroxiredoxin 4 measurement, were performed.

Results: There was a tendency towards lower peroxiredoxin 4 levels in non-obese PCOS subjects (5674.8 ± 3822.4 pg/ml), higher in obese PCOS (6588.9 ± 3731.0 pg/ml) and even higher in obese patients without PCOS (7724.6 ± 4840.4 pg/ml). Patients with abdominal obesity according to waist circumference and waist-to-hip ratio had significantly higher levels of peroxiredoxin compared to those without (7108.2 ± 4568.0 vs. 5079.8 ± 2555.4 pg/ml; p = 0.015 and 7310.6 ± 2646.2 vs. 4785.0 ± 2646.2 pg/ml; p = 0.013). There was no difference in peroxiredoxin 4 levels in patients with and without insulin resistance, hypertension, dislipidemia, hyperandrogenemia, metabolic syndrome. Peroxiredoxin 4 showed weak positive correlation to weight (r = 0.228; p = 0.044) and visceral adiposity index (r = 0.278; p = 0.031) and higher to erythrocyte sedimentation rate (r = 0.4; p < 0.01), but not to hormonal parameters and insulin sensitivity indexes.

Conclusions: Non-obese patients with PCOS have a tendency towards lower peroxiredoxin 4 levels compared to obese patients with and without PCOS. Patients with abdominal obesity have significantly higher peroxiredoxin 4 levels than those without. We were not able to prove correlation between peroxiredoxin 4 levels and hormonal and carbohydrate status of the PCOS patients.
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http://dx.doi.org/10.1016/j.jogoh.2019.04.002DOI Listing
November 2019

Higher levels of thioredoxin interacting protein (TXNIP) in patients with prediabetes compared to obese normoglycemic subjects.

Diabetes Metab Syndr 2019 Jan - Feb;13(1):734-737. Epub 2018 Nov 30.

Clinic of Endocrinology, University Hospital "Alexandrovska", Medical University-Sofia, 1 GeorgiSofiiski Str., 1431, Sofia, Bulgaria.

Background: Thioredoxin interacting protein (TXNIP) is one of the mediators of oxidative stress induced beta-cell glucotoxisity. TXNIP might play a key role in impaired glucose homeostasis preceding overt T2DM. The aim of the present study was to compare TXNIP levels between patients with prediabetes and obese normoglycemic controls and to evaluate the link between TXNIP and metabolic risk factors.

Patients And Methods: In the present study we included 79 patients with mean age 50.3 ± 10.6 years, divided into two age and BMI matched groups -control group with obesity without glycemic disturbances (NGT) (n = 40) and prediabetes (n = 39).

Results: We found significantly higher levels of TXNIP in patients with prediabetes compared to normoglycemic obese controls (54.2 ± 69.9 vs. 23.9 ± 47.1 pg/ml; p = 0.03). The levels of TXNIP gradually increased from normal glucose tolerance trough IFG/IGT only to IFG + IGT (27,1; 44.0; 49.9 and 95.7 pg/ml respectively; p = 0.025 between NGT and IFG + IGT). TXNIP levels correlated weakly only with fasting blood glucose (r = 0.235; p = 0.04) but not with glucose during OGTT or the markers of insulin resistance.

Conclusions: The levels of TXNIP are higher in patients with prediabetes compared to normoglycemic controls as they increase gradually from NGT trough IFG/IGT only to IFG + IGT.
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http://dx.doi.org/10.1016/j.dsx.2018.11.056DOI Listing
May 2019

Higher levels of IL-18 in patients with prediabetes compared to obese normoglycaemic controls.

Arch Physiol Biochem 2020 Dec 11;126(5):449-452. Epub 2019 Jan 11.

Laboratory of Tumor Immunology and Cell Therapy, Hyogo College of Medicine, Tokyo, Japan.

: Overweight and obesity are linked to low-grade chronic inflammation that can impair normal insulin function and induce insulin resistance. The aim of this study was to compare IL-18 levels between patients with prediabetes and obese normoglycaemic controls.: In this study, we included 131 patients with mean age 54.9 ± 9.1 years, divided into two groups - group 1 with obesity without glycaemic disturbances ( = 66) and group 2 with prediabetes ( = 65). IL-18 was measured using enzyme-linked immunosorbent assay (ELISA) method.: Patients with prediabetes had significantly higher levels of IL-18 compared to obese controls (304.0 ± 220.4 vs. 233.6 ± 103.6 pg/l, =.029). When patients with prediabetes were divided into IFG only, IGT only and IFG + IGT the highest levels of IL-18 were found in IGT only patients.: Patients with prediabetes have higher levels of IL18 compared to obese normoglycemic controls.
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http://dx.doi.org/10.1080/13813455.2018.1555667DOI Listing
December 2020

Cultured Cells Isolated from CNS Indolent B-Cell Lymphoma Have Characteristics of Mesenchymal Stem Cells: A Clinical Case and Scientific Research.

Int J Hematol Oncol Stem Cell Res 2018 Jul;12(3):169-174

Laboratory of Clinical Immunology, University Hospital "St. Ivan Rilski", Department of Clinical Laboratory and Clinical Immunology, Medical University of Sofia, Sofia, Bulgaria.

The case report presented here describes the culturing and characterization of mesenchymal stem cells (MSCs) isolated from a primary indolent B-cell lymphoma, located in the CNS of an immunocompetent patient. The presence of such cells in the tumor mass can further elucidate the pathogenesis of the disease and reveal possible future approaches for its treatment. We present a case report of a 61-year-old immunocompetent woman who had an episode of confusion with numbness in the right leg and the right arm, slurred and dysarthric speech and urine incontinence. The peripheral blood tests were normal. The neurological examination demonstrated a latent hemi-paresis of the right side, aphasia, discrete hypertension and bradypsychia. The ophthalmologic examination revealed left quadranopsia. Computed tomography and magnetic resonance imaging of the brain showed a 3.5 × 2.9 cm infiltrative neoplastic lesion involving the left temporal parenchyma. The morphological features and the immunophenotyping of the lymphoid cell composition were consistent with low-grade (indolent) B-lymphocyte non-Hodgkin's lymphoma of CNS. Cells, isolated from the resected tumor mass, were cultured in medium containing 10% fetal bovine serum (FBS) and characterized by their morphology, growth, phenotype, clonogenicity and osteogenic differentiation. It was apparent that the cultured cells isolated from the indolent B- cell lymphoma located in the CNS have the basic characteristics of mesenchymal stem cells. The presence of MSCs is described for the very first time in such type of tumor. The well-known immunosuppressive properties of the MSCs may represent another mechanism favouring the tumor growth.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6305270PMC
July 2018

Interleukin-18 serum level is elevated in type 2 diabetes and latent autoimmune diabetes.

Endocr Connect 2018 Jan 7;7(1):179-185. Epub 2017 Dec 7.

Laboratory of Tumor Immunology and Cell TherapyHyogo College of Medicine, Hyogo, Japan.

Background: Interleukin-18 (IL-18) is an inflammatory cytokine found to be elevated in obesity, metabolic syndrome and type 2 diabetes (T2D) as a part of the chronic low-grade inflammatory process in these states. The aim of the study was to evaluate the interleukin level in patients with latent autoimmune diabetes of the adults (LADA) in comparison to that in T2D subjects.

Materials And Methods: IL-18 was analyzed through enzyme-linked immunosorbent assay in 76 participants with T2D and 24 with LADA and 14 control subjects. Evaluation was also carried out in body mass index (BMI)- and glycemic control-matched diabetic patients.

Results: The serum concentration of IL-18 was higher in patients with T2D (389.04 ± 203.44 pg/mL) and LADA (327.04 ± 144.48 pg/mL) than that in control subjects (219.88 ± 91.03 pg/mL),  < 0.05. However, it was not significantly different between both diabetic groups ( = 0.255) despite higher IL-6 (4.78 ± 5.84 vs 1.79 ± 0.96 pg/mL,  < 0.001) and hs-CRP (2.60 ± 1.70 vs 1.29 ± 1.20 mg/L,  = 0.002) level in T2D patients. The results were persistent in BMI-matched subjects with diabetes (IL-18 = 403.48 ± 226.32 vs 329.30 ± 146.30 pg/mL, respectively for T2D and LADA,  = 0.391). The correlations in T2D group concerning HDL cholesterol ( = -0.377,  = 0.001), postprandial glucose ( = 0.244,  = 0.043), IL-6 ( = 0.398,  < 0.001) and hs-CRP ( = 0.427,  = 0.001) were not confirmed in LADA and control subjects.

Conclusion: The IL-18 serum level was higher in T2D and LADA than that in control subjects, but did not differ between both diabetic groups, even when they were BMI matched. Correlations with lipid, glycemic and inflammatory parameters were present in T2D only.
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http://dx.doi.org/10.1530/EC-17-0273DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5776671PMC
January 2018
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