Wieslaw Zarzycki

Metabolic fingerprint of Gestational Diabetes Mellitus.

UNLABELLED Gestational Diabetes (GDM) is causing severe short- and long-term complications for mother, fetus or neonate. As yet, the metabolic alterations that are specific for the development of GDM have not been fully determined, which also precludes the early diagnosis and prognosis of this pathology. In this pilot study, we determine the metabolic fingerprint, using a multiplatform LC-QTOF/MS, GC-Q/MS and CE-TOF/MS system, of plasma and urine samples of 20 women with GDM and 20 with normal glucose tolerance in the second trimester of pregnancy. Plasma fingerprints allowed for the discrimination of GDM pregnant women from controls. In particular, lysoglycerophospholipids showed a close association with the glycemic state of the women. In addition, we identified some metabolites with a strong discriminative power, such as LPE(20:1), (20:2), (22:4); LPC(18:2), (20:4), (20:5); LPI(18:2), (20:4); LPS(20:0) and LPA(18:2), as well as taurine-bile acids and long-chain polyunsaturated fatty acid derivatives. Finally, we provide evidence for the implication of these compounds in metabolic routes, indicative of low-grade inflammation and altered redox-balance, that may be related with the specific pathophysiological context of the genesis of GDM. This highlights their potential use as prognostic markers for the identification of women at risk to develop severe glucose intolerance during pregnancy. BIOLOGICAL SIGNIFICANCE Gestational Diabetes Mellitus (GDM) is increasing worldwide and, although diabetes usually remits after pregnancy, women with GDM have a high risk of developing postpartum type 2-diabetes, particularly when accompanied by obesity. Therefore, understanding the pathophysiology of GDM, as well as the identification of potentially modifiable risk factors and early diagnostic markers for GDM are relevant issues. In the present study, we devised a multiplatform metabolic fingerprinting approach to obtain a comprehensive picture of the early metabolic alternations that occur in GDM, and may reflect on the specific pathophysiological context of the disease. Future studies at later stages of gestation will allow us to validate the discriminant power of the identified metabolites.

The Effect of Intragastric Balloon on Plasma Ghrelin, Leptin, and Adiponectin Levels in Patients with Morbid Obesity

CONTEXT Ghrelin and leptin are hormones regulating appetite and metabolic processes. Adiponectin plays an important role in the modulation of glucose and lipid metabolism. OBJECTIVE The objective of the study was to evaluate the levels of plasma ghrelin, leptin, and adiponectin in obese subjects treated with bioenterics intragastric balloon (BIB), low-calorie diet (1500 kcal), and physical exercise. DESIGN BIB was placed for 6 months in 21 subjects with body mass index 47.3 +/- 5.7. The control group consisted of 15 morbidly obese subjects treated with a low-calorie diet and physical effort. Plasma hormone levels were determined by RIA. RESULTS In the BIB group, the insertion of the balloon caused a considerable reduction in body mass over a 6-month period (17.1 +/- 8.0 kg) as compared with the control group (3.2 +/- 6.4 kg). After 1 month, the levels of ghrelin increased from 621.9 +/- 182.4 to 903.9 +/- 237 pg/ml and thereafter gradually decreased, reaching the starting level 3 months after the removal of the balloon. In the same group, the levels of leptin decreased from 61.3 +/- 36.7 to 39.9 +/- 17.5 ng/ml. In the control group, the corresponding levels of ghrelin and leptin remained relatively stable. During the observation period, in the BIB group, the levels of adiponectin remained unchanged as opposed to a transient increase noted in the control group. CONCLUSIONS In patients with morbid obesity, weight loss induced by BIB is associated with a decrease in plasma leptin and a transient elevation of plasma ghrelin. It is likely that the changes in hormones regulating the energy balance caused by BIB can prevent an increase in adiponectin level.

Microvascular abnormalities in capillaroscopy correlate with higher serum IL-18 and sE-selectin levels in patients with type 1 diabetes complicated by microangiopathy

Microvascular abnormalities are one of the most important causes of persistent diabetic complications. The aim of this study was to compare microvascular changes examined by nailfold capillaroscopy with serum concentrations of soluble E-selectin (sE-selectin) and IL-18 in type 1 diabetic patients with and without microangiopathy. Serum levels of sE-selectin and IL-18 were determined by an enzyme-linked immunosorbent assay in 106 patients with type 1 diabetes and in 40 healthy controls. All diabetic patients were evaluated by extensive clinical, laboratory and capillaroscopic studies. Morphological changes were observed by nailfold capillaroscopy in 86 out of 106 (81%) diabetic patients. Severe capillaroscopic changes were seen in 32 out of 54 (59%) patients with microangiopathy, but in only seven out of 52 (13%) patients without microangiopathy. Higher serum levels of sE-selectin (p < 0.001) and IL-18 (p < 0.05) were demonstrated in diabetic patients compared to controls. Significant differences of sE-selectin (p , 0.001) and IL-18 (p < 0.01) serum concentrations were observed between diabetic patients with microangiopathy and controls. Moreover, comparison between patients with and without microangiopathic complications showed a significantly higher capillaroscopic score and sE-selectin serum concentration in the group with microangiopathy (p < 0.001). Furthermore, diabetic patients with severe microvascular changes in capillaroscopy showed significantly higher IL-18 (p < 0.001) and sE-selectin (p < 0.05) serum levels than subgroups without changes or with mild abnormalities. Our findings suggest that abnormalities in nailfold capillaroscopy may reflect the extent of microvascular involvement and are associated with higher sE-selectin and IL-18 serum levels, as well as with microangiopathic complications in diabetic patients.

Optimization and validation of a chiral GC–MS method for the determination of free d-amino acids ratio in human urine: Application to a Gestational Diabetes Mellitus study

Gestational Diabetes Mellitus (GDM) is defined as glucose intolerance with onset or first recognition during pregnancy. It is affecting approximately up to 14% of all pregnancies with an increasing tendency. GDM has been related to relevant short-term and long-term health complications for both mother and offspring. Recent studies strongly emphasized the role of several essential amino acids in the pathogenesis of obesity and highlighted their strong correlation with insulin resistance, but there are no references related to modifications in D-AAs in biological fluids. As D-AA elimination proceeds mainly by renal excretion, urine was the selected sample to evaluate the alterations in free D-AAs ratio in a GDM study. Only 1 mL of first void urine or standard solution was required for purification, by using a Discovery DSC-SCX SPE cartridge (500 mg/3 mL) and derivatization into their N(O)-pentafluoropropionyl amino acid 2-propyl esters. Enantiomeric separation was carried out by GC-MS on a Chirasil-L-Val N-propionyl-L-valine-tert-butylamide polysiloxane fused-silica capillary column (25 m×0.25 mm I.D., 0.12 μm film thickness, Agilent Technologies, Waldbronn, Germany), under programmed temperature elution. Detection was performed with an ion trap mass analyzer, operating in the full scan mode in the m/z 50-350 range. 14 pairs of derivatives of D-and L-AAs were separated. The steps of sample preparation, derivatization and GC-MS conditions were optimized for both urine and standards. Several conditions affecting the SPE procedure, such as sorbent mass/volume ratio of the cartridge, sample dilution and pH, were optimized. Volume of reagents and solvents and reaction temperature and time were also tested for the derivatization. Regarding the GC-MS parameters, split ratio, temperature program and mass range were optimized. The final method was validated in terms of linearity, sensitivity, accuracy and precision for D-Ala, D-Pro, D-Ser, D-Met, D-Phe, D-Glu, D-Orn and D-Lys. Identification of AAs in urine samples was based on retention time and mass spectra. Urine from 20 women with GDM and 20 pregnant women with normal glucose tolerance (after 2-h 75-g oral glucose tolerance test), matched according to the week of gestation and age (22-28 week of gestation and age 24-37 years), were enrolled into the study. %D-Relative amounts were determined for Ala, Val, Thr, Ser, Leu, Asx (Asp+Asn), Glx (Glu+Gln), Met, Phe, Tyr, Orn and Lys. Statistically significant differences (p<0.05) were observed only for D-Phe and higher values were found in the GDM group. It is possible that D-Phe could be involved in metabolic/signaling pathways to compensate early stages of insulin resistance, although further work is necessary to confirm this hypothesis.

Median nerve conduction impairment in patients with diabetes and its impact on patients' perception of health condition: a quantitative study.

IntroductionImpaired mobility and compromised manual dexterity leading to difficulties with the activities of daily living (ADL) are an inherent part of the clinical picture in diabetes. Hand function in diabetes is influenced by a variety of pathologies: the median nerve, the most important nerve of the hand, can suffer from metabolic disturbances, ischemia and/or entrapment neuropathies. The resulting deterioration in functional capacity is likely to have significant consequences for the ability to perform ADL, influencing adjustment to diabetes and affecting quality of life. The aim of the present study was to examine the influence of hand function as measured by median motor nerve conduction on quality of life, taking into account various aspects of functioning in patients with diabetes, including activities of daily living, psychological status and acceptance of illness.Patients and methodsSeventy one hospital patients with diabetes participated in the study. Electrophysiological recordings of conductance in the median nerve were obtained for both hands and the relationship between hand function and functional status (BI), depression and anxiety (HADS), adjustment to illness (AIS) and their effect on quality of life (SF-36v2 and QLI) was studied.ResultsDamage to the median nerve of the left hand was associated with significant differences in functioning in the physical, but not the mental component of the SF-36v2, p = 0.03 and in functional status (p = 0.006). QOL was associated with depression, patient age, acceptance of illness, functional ability and to a small, but significant extent with median nerve damage to the right hand on the measure of conduction velocities (R2 =0.726).ConclusionsNerve conductance studies demonstrated a small, but significant effect of hand function on quality of life. Impairment of the median nerve in the left hand was associated with functional difficulties in the activities of daily living and a diminished quality of life in the area of physical functioning. No dependencies of this kind were found for the right hand, which may reflect the greater compensatory capacity of the right hand resulting from improved efficiency due to practice.

GC–MS based Gestational Diabetes Mellitus longitudinal study: Identification of 2-and 3-hydroxybutyrate as potential prognostic biomarkers

HIGHLIGHTSGC–MS offers high‐throughput molecular profiling to study metabolic signature of diabetes.Metabolomics may serve for early prediction of GDM and developing T2DM.2‐hydroxybutyrate and 3‐hydroxybutyrate may be potential prognostic biomarkers.Metabolomics is bringing knowledge gaps in current management of diabetes.Translation of metabolomics research for clinical practice is desired. ABSTRACT Gestational Diabetes Mellitus (GDM) causes severe short‐ and long‐term complications for the mother, fetus and neonate, including type 2‐diabetes (T2DM) later in life. In this pilot study, GC–Q/MS analysis was applied for plasma metabolomics fingerprinting of 24 healthy and 24 women with GDM at different stages of gestation (second and third trimester) and postpartum (one and three months). Multivariate (unsupervised and supervised) statistical analysis was performed to investigate variance in the data, identify outliers and for unbiased assessment of data quality. Plasma fingerprints allowed for the discrimination of GDM pregnant women from controls both in the 2nd and 3rd trimesters of gestation. However, metabolic profiles tended to be similar after delivery. Follow up of these women revealed that 4 of them developed T2DM within 2 years postpartum. Multivariate PLS‐DA models limited to women with GDM showed clear separation 3 months postpartum. In the 2nd trimester of gestation there was also a clear separation between GDM women that were normoglycemic after pregnancy and those with recognized postpartum T2DM. Metabolites that had the strongest discriminative power between these groups in the 2nd trimester of gestation were 2‐hydroxybutyrate, 3‐hydroxybutyrate, and stearic acid. We have described, that early GDM comprises metabotypes that are associated with the risk of future complications, including postpartum T2DM. In this pilot study, we provide evidence that 2‐hydroxybutyrate and 3‐hydroxybutyrate may be considered as future prognostic biomarkers to predict the onset of diabetic complications in women with gestational diabetes after delivery.

Clinical usefulness of videocapillaroscopy and selected endothelial cell activation markers in people with Type 1 diabetes mellitus complicated by microangiopathy

PURPOSE Microvascular abnormalities are one of the most important causes of persistent diabetic complications. The aim of our study was to compare microvascular changes examined by nailfold videocapillaroscopy (NVC) examination with serum concentrations of vascular endothelial growth factor (VEGF), soluble thrombomodulin (sTM) and endothelin-1 (ET-1) in people with Type 1 diabetes with and without microangiopathy. MATERIAL/METHODS The study included 106 people with Type 1 diabetes and 40 healthy controls. All participants were evaluated by extensive clinical, laboratory and capillaroscopic studies. NVC was performed using a stereomicroscope SZ 4045 (Olympus, Germany). The intensity of morphological changes was graded from 0 to 3. Serum levels of VEGF, sTM and ET-1 were determined by an enzyme-linked immunosorbent assay (ELISA). RESULTS Morphological changes were observed by NVC in 86 out of 106 (81%) people with Type 1 diabetes mellitus. Severe capillaroscopic changes (score 3) were seen in 32 out of 54 (59%) people with microangiopathy, but in only seven out of 52 (13%) individuals without microangiopathy. Higher serum concentration of VEGF (p<0.001), ET-1 (p<0.001) and sTM (p<0.05) were demonstrated in people with diabetes complicated with microangiopathy compared to healthy controls. Moreover, comparison between people with and without microangiopathic complications showed a significantly higher capillaroscopic score and sTM serum concentration in the group with retinopathy (p<0.001) nephropathy (p<0.001) and neuropathy (p<0.01). CONCLUSIONS Our results suggest that abnormalities in NVC may reflect the extent of microvascular involvement and associated with higher VEGF, sTM and ET-1 serum levels, as well as with microangiopathic complications in diabetic people.

N-Acetyl-β-D-hexosaminidase in gestational diabetes mellitus - a preliminary study.

PURPOSE N-Acetyl-β-D-hexosaminidase (HEX) is an exoglycosidase which has been extensively studied and which has been used as a marker for inflammation. It was therefore thought that measurement of the activity of this enzyme might be useful in diagnosing gestational diabetes mellitus (GDM) as this condition is frequently associated with inflammation. The main object of the study was the determination of N-acetyl-β-D-hexosaminidase activity in women with GDM and 3 months postpartum in comparison with control groups of non-pregnant and healthy pregnant women. MATERIAL AND METHODS Twenty-five blood serum samples from women with GDM and women 3 months postpartum; 20 blood serum samples from non-pregnant and healthy pregnant women (control groups) were enrolled into the study. Serum was prepared from all blood samples and HEX activity was measured by the method of Chateriee et al. (modified by Zwierz et al). RESULTS A statistically significantly increase in the activity of HEX in the GDM blood serum was found as compared to the control groups (p<0.05). Further analysis showed a statistically significant decrease in the activity of HEX among postpartum women, but the level of enzyme activity was still above the normal control level in comparison to the control group of nonpregnant healthy women (p<0.05). CONCLUSIONS Changes in the activity of HEX appear to be involved in the pathogenesis of gestational diabetes mellitus. Determination of HEX activity may have prognostic significance as an early indicator of diabetes mellitus among GDM women in the future.