Exosome Proteomics Reveals the Deregulation of Coagulation, Complement and Lipid Metabolism Proteins in Gestational Diabetes Mellitus.

Exosomes are small extracellular vesicles with a variable protein cargo in consonance with cell origin and pathophysiological conditions. Gestational diabetes mellitus (GDM) is characterized by different levels of chronic low-grade inflammation and vascular dysfunction; however, there are few data characterizing the serum exosomal protein cargo of GDM patients and associated signaling pathways. Eighteen pregnant women were enrolled in the study: 8 controls (CG) and 10 patients with GDM. Blood samples were collected from patients, for exosomes' concentration. Protein abundance alterations were demonstrated by relative mass spectrometric analysis and their association with clinical parameters in GDM patients was performed using Pearson's correlation analysis. The proteomics analysis revealed 78 significantly altered proteins when comparing GDM to CG, related to complement and coagulation cascades, platelet activation, prothrombotic factors and cholesterol metabolism. Down-regulation of Complement C3 (C3), Complement C5 (C5), C4-B (C4B), C4b-binding protein beta chain (C4BPB) and C4b-binding protein alpha chain (C4BPA), and up-regulation of C7, C9 and F12 were found in GDM. Our data indicated significant correlations between factors involved in the pathogenesis of GDM and clinical parameters that may improve the understanding of GDM pathophysiology. Data are available via ProteomeXchange with identifier PXD035673.

Interleukin-8, CXCL10, CXCL11 and their role in insulin resistance in adult females with subclinical hypothyroidism and prediabetes.

In obesity, the hormonal secretion of the thyroid gland switches from homeostasis to type 2 allostasis in order to adapt to persistent modifications of adipose tissue and inflammation. Previous meta-analyses have linked obesity with an increased risk of developing thyroid diseases, prediabetes, and type 2 diabetes mellitus. We designed an observational cross-sectional study including all female patients presenting consecutively in an ambulatory clinic for 16 months. This study aimed to describe the level of serum cytokines and chemokines in relation to TSH, fT4 and insulin resistance (IR) indexes in patients with subclinical hypothyroidism (SCH). The study included 72 women with a median age of 59 ± 17.75 years, and a mean BMI (Body Mass Index) of 31.48 ± 6.75 kg/m2. Modelling homeostasis model assessment of IR indices (HOMA-IR) based on chemokines (IL-8, CXCL10, CXCL11, leptin), C-reactive protein, the presence or absence of SCH, taking into account age, BMI, abdominal circumference, glycated haemoglobin (HbA1c), and anti-thyroid peroxidase antibodies (ATPO) as covariates, identified a single chemokine that was significantly associated with the dependent variable (IL-8). IR indices are negatively associated with IL-8 in female patients with subclinical hypothyroidism, but the effect of the cytokine is minimal. BMI rather than TSH influences the level of CXCL11 in our population. CXCL10 has a tendency to increase in patients with SCH, obesity and prediabetes, with no association with TSH.

Interleukins and redox impairment in type 2 diabetes mellitus: mini-review and pilot study.

BACKGROUND: Type 2 diabetes mellitus (T2DM) represents a leading cause of morbidity and premature mortality, low-grade inflammation being acknowledged as a key contributor to its development and progression. A tailored therapeutic approach, based on sensitive and specific biomarkers, could allow a more accurate analysis of disease susceptibility/prognostic and of the response to treatment. OBJECTIVES: This mini-review and pilot study had two main goals: (1) reviewing the most recent literature encompassing the use of interleukins as inflammatory markers influenced by the redox imbalances in T2DM and (2) assessing parameters that conjunctly evaluate the redox impairment and inflammatory burden of T2DM patients, taking into consideration smoking status, as such group-specific biomarkers are scarcely reported in literature. METHODS: Firstly, PubMed database was surveyed to select and review the relevant studies employing interleukins as T2DM biomarkers and to assess if studies using combined inflammatory-redox indices were reported. Then, routine biochemical parameters were assessed in a pilot study -T2DM patients with 3 subgroups: non-smokers, smokers and ex-smokers, were compared to a control group of non-diabetic, apparently healthy non-smokers. Protein (AOPPs, AGEs), lipid/HDL (Amplex Red-based method) oxidative damage and inflammatory status (CRP, IL-1ß, IL-6, IL-10) biomarkers were assessed. Cytokine ratios and 2 oxidative-inflammatory status indices were developed (IH1 and IH2) and evaluated. RESULTS: We observed significant differences in terms of serum redox and inflammatory status (AOPPs, AGEs, CRP, CRP/HDL, CRP/IL-6, IL-10/IL-6, IH1) between T2DM patients compared to control and, moreover, between the subgroups formed considering smoking status (CRP, CRP/HDL, IH1). Glycemic control strongly influenced inflammatory status biomarkers: glycemia was positively correlated with the inflammatory parameters (CRP/IL-10) and inversely with the anti-inflammatory ones (IL-10, IL-10/IL-1ß ratio). CONCLUSIONS: Several of the assessed parameters may possess prognostic value for diabetics, especially when comparing subgroups with a different smoking history and could prove useful in clinical practice for assessing disease progress and therapeutic efficacy.

Zinc status, insulin resistance and glycoxidative stress in elderly subjects with type 2 diabetes mellitus.

Zinc deficiencies have been reported in numerous pathologies, such as diabetes mellitus, but also in the physiological process of ageing. Similarly, the end products of glycoxidation processes, advanced glycation end products (AGEs), are damaging compounds, a myriad of reports linking them to the development and progression of several age-associated chronic diseases. The aim of the present study was to analyze the relationships between zinc status, glycoxidative stress and insulin resistance (IR) in elderly subjects with type 2 diabetes mellitus (T2DM). A group of 52 non-smoking subjects (9 men and 43 women, aged 65-83 years) were enrolled in this cross-sectional study: 27 patients with T2DM, and 25 apparently healthy control subjects. Serum zinc (Zn) levels were assessed using a commercial kit based on an end-point colorimetric method, and serum AGEs were evaluated with a fluorimetric analytic procedure. The calculated glucose-to-zinc ratio (Gly/Zn), insulin-to-zinc ratio (Ins/Zn) and insulin-zinc resistance index (HOMA-IR/Zn) were further used to study the associations between serum Zn levels, secretory function of ß-pancreatic cells and AGEs. T2DM patients presented significantly higher serum insulin and Zn levels, as compared to the controls. We found a significant inverse correlation between Zn and AGEs, and a strong positive correlation between AGEs and the Gly/Zn ratio, suggesting that both Zn and AGEs are biomarkers that could reflect the persistence of hyperglycemia. We identified new surrogate biomarkers useful for the assessment of glycemic control with great potential for the development of preventive and therapeutic strategies for elderly diabetics, based on the evaluation of serum Zn levels.

Subclinical hypothyroidism has no association with insulin resistance indices in adult females: A case-control study.

Longitudinal studies have indicated an association between thyroid function and insulin resistance (IR) or a neutral relationship. Both the lowest tertile of free thyroxine (fT4) and the highest tertile of free triiodothyronine (fT3) were found to be associated with IR in cross-sectional studies. The aim of the present study was to analyze the association between IR and subclinical hypothyroidism in a female adult population from Bucharest, Romania. This is a retrospective pilot case-control study that included female patients examined by two endocrinologists and a diabetologist in an outpatient clinic. The retrospective follow-up had a one-year duration and included the evaluation of thyroid function tests and IR indices based on fasting insulinemia and C-peptide. The study included 176 women, 91 with subclinical hypothyroidism, with a median age of 60±17 years and a mean body mass index (BMI) of 27.79±4.76 kg/m2. The majority of the population (50%) was diagnosed with autoimmune thyroiditis, and 17.05% with goitre. The univariate logistic regression using hypothyroidism as the explaining variable found no evidence of a significant relationship between a decreased thyroid function and IR (OR 1.32; P=0.36). Metabolic syndrome was probably the most important determinant of IR in the population group studied. Thus, it was not the thyroid function per se, but the coexistence of other elements of this syndrome that prevailed in determining IR. Advantages to the study are the design that permitted evaluation of IR and the thyroid function at different moments in time as well as the uniformity of the blood tests. The multivariate analyses were adjusted for age, lipid profile and treatment; however, one limiting factor was the absence of other hormonal blood tests. In summary, there was no association between the thyroid function tests (TSH, fT4) and IR indices in adult Romanian women in a case-control study with one-year retrospective follow-up.

No Association between 25-Hydroxyvitamin D and Insulin Resistance or Thyroid Hormone Concentrations in a Romanian Observational Study.

Background and objectives: Vitamin D is involved in insulin resistance through genomic and non-genomic mechanisms. Several observational and randomized studies have discrepant results; some of them showed an improved insulin resistance (IR), and others a neutral effect after vitamin D deficiency is corrected. Materials and Methods: We designed a retrospective observational study that included all women who presented for 33 months in an outpatient clinic in Bucharest, Romania. Results: We analyzed 353 patients with a mean age of 58.5 ± 13.7 years, a mean body mass index (BMI) of 27.36 ± 4.87 kg/m-2, and a mean level of 25-hydroxyvitamin D (25OHD) of 39.53 ± 15.73 ng/mL. There were no differences in the calculated Homeostatic Model Assessment of Insulin Resistance variants 1 and 2 (HOMA-IR) and the Quantitative Insulin Sensitivity Check Index (QUICKI) between women with vitamin D deficit versus normal values. In multivariate analysis, there was no significant relation between 25OHD and the response variables considered by us. Conclusions: We observed a small positive correlation between a higher level of 25OHD and increased glycosylated hemolobin (HbA1c) or IR indices without clinical significance. Other modifiable or non-modifiable factors override 25OHD influence on IR in adult women with a normal serum level and may contribute to the remainder of the variability observed.

Spectrophotometric versus spectrofluorometric assessment in the study of the relationships between lipid peroxidation and metabolic dysregulation.

Reactive oxygen species are crucial to normal cell function, but are also part of the pathogenesis of multiple modern maladies. As such, sensitive, fast, and reliable methods of appreciating redox status are needed. We aimed to optimize the Amplex Red (AR) and ferric-xylenol orange (FOX) methods using human serum samples, rat tissue homogenates, and mitochondrial preparations. For AR, we intended to reduce probe concentration, maintaining method sensitivity, as well as extending its use from isolated lipoproteins samples, and readjust it for a high-throughput application. Also, we evaluated the usefulness of a modified xylenol orange-based spectrophotometric protocol, comparing and contrasting these methods in terms of clinical relevance and suitability for their further use in assessing redox status of various biological samples in different pathological conditions. Our results show that these optimized protocols are suitable for complex in vivo studies, as they require low quantities of sample and reagents, and are sensitive, rapid, and economical, with the option of adapting them for high-throughput analysis. For a better assessment of oxidative status of serum-derived samples, the two methods can be used concurrently, while for tissue-derived ones, either can be employed for the measurement of a global redox status.