Irisin levels in LMNA-associated partial lipodystrophies.

AIM: The adipo-myokine irisin regulates energy expenditure and fat metabolism. LMNA-associated familial partial lipodystrophy (FPLD2) comprises insulin resistance, muscle hypertrophy and lipoatrophy. The aim of this study was to investigate whether irisin could be a biomarker of FPLD2. PATIENTS AND METHODS: This case control study included 19 FPLD2 subjects, 13 obese non-diabetic (OND) patients and 19 healthy controls (HC) of normal weight (median BMI: 26, 39 and 22 kg/m2, respectively). Serum irisin and leptin levels, body composition (DXA/MRI) and metabolic/inflammatory parameters were compared in these three groups. RESULTS: BMI and MRI intra-abdominal fat significantly differed among these three groups, whereas DXA total fat mass and leptin levels were higher in the OND group, but did not differ between HC and FPLD2. Lipodystrophy patients had higher intra-abdominal/total abdominal fat ratios than the other two groups. Irisin levels were higher in FPLD2 and OND patients than in HC (medians: 944, 934 and 804 ng/mL, respectively). However, irisin/leptin ratios and lean body mass percentages were strikingly higher, and lean mass indices lower, in FPLD2 and HC than in the OND (median irisin/leptin ratios: 137, 166 and 21, respectively). In the entire study group, irisin levels positively correlated with BMI, lean body mass and index, intra-abdominal/total abdominal fat ratio, triglyceride, cholesterol, insulin, glucose and HbA1c levels. Also, intra-abdominal/total abdominal fat ratio and lean body mass better differentiated the three groups only in female patients. CONCLUSION: Circulating irisin is similarly increased in FPLD2 and OND patients, who are characterized by higher lean body mass regardless of their clearly different fat mass. However, irisin/leptin ratios, strikingly higher in FPLD2 than in OND patients, could help to make the diagnosis and prompt genetic testing in clinically atypical cases.

Purity of islet preparations and 5-year metabolic outcome of allogenic islet transplantation.

In allogenic islet transplantation (IT), high purity of islet preparations and low contamination by nonislet cells are generally favored. The aim of the present study was to analyze the relation between the purity of transplanted preparations and graft function during 5 years post-IT. Twenty-four patients with type 1 diabetes, followed for 5 years after IT, were enrolled. Metabolic parameters and daily insulin requirements were compared between patients who received islet preparations with a mean purity <50% (LOW purity) or ≥50% (HIGH purity). We also analyzed blood levels of carbohydrate antigen 19-9 (CA 19-9)-a biomarker of pancreatic ductal cells-and glucagon, before and after IT. At 5 years, mean hemoglobin A1c (HbA1c levels) (P = .01) and daily insulin requirements (P = .03) were lower in the LOW purity group. Insulin independence was more frequent in the LOW purity group (P < .05). CA19-9 and glucagon levels increased post-IT (P < .0001) and were inversely correlated with the degree of purity. Overall, our results suggest that nonislet cells have a beneficial effect on long-term islet graft function, possibly through ductal-to-endocrine cell differentiation. ClinicalTrial.gov NCT00446264 and NCT01123187.

Reversed-phase liquid chromatography column testing and classification: physicochemical interpretation based on a wide set of stationary phases.

The high number of stationary phases commercially available for liquid chromatography makes the choice of the analyst a real headache. In order to provide a tool to carry out this choice on objective basis, the present work proposes interpretations of the column classifications obtained, thanks to a previously described testing procedure. The meaning of principal components was attributed to crossing over information carried by loading plots and groups revealed by hierarchical cluster analysis (HCA) on the corresponding score plots. At high solvent ratio, the retention seemed to be governed by enthalpy, whereas at low solvent ratio, entropic phenomena were predominating. Finally, the behavior of known families of RPLC columns was studied giving rise either to homogeneous groups like polar embedded grafts columns or to scattered families like Aqua type columns.

Reversed-phase liquid chromatography testing. Role of organic solvent through an extended set of columns.

Column testing is a primary concerns for analysts. It is of use not only for the choice of set of development columns with different behaviors, but also for a substitution column in a validated method or as a quality control of new batches of stationary phase. A validated chromatographic procedure for column testing was applied to 42 commercially available columns, including alkyl, polar embedded and Aqua type stationary phases. This procedure was based on the use of two different solvents: MeOH and MeCN; and two different solvent/aqueous buffer fractions. Principal component analysis has been combined to hierarchical cluster analysis to provide both rational and objective classifications. The solvent effects were then studied on the obtained representations, revealing the necessity for considering both the solvent nature and its fraction in RPLC column testing. Differences observed depending on the solvent nature and fractions revealed quite different chromatographic behaviors according to these parameters.

Testing of "special base" columns in reversed-phase liquid chromatography. A rational approach considering solvent effects.

A methodology for building a chromatographic test aiming at characterizing special base stationary phases was described. Instead of choosing its conditions a priori, a "full" comprehensive test based on extended running conditions was performed on a 12 column set. The conditions were carefully chosen from their ability to take into account the solvent and the pH effects. Principal component analysis (PCA) has been combined to hierarchical cluster analysis both to provide interpreted classifications and to reduce drastically the test itself by eliminating redundant information. The final reduced test can be considered optimal because the minimized set of test conditions allows to provide as much information as in the initial full test.

Reversed-phase liquid chromatography column testing: robustness study of the test.

Choosing the right RPLC column for an actual separation among the more than 600 commercially available ones still represents a real challenge for the analyst particularly when basic solutes are involved. Many tests dedicated to the characterization and the classification of stationary phases have been proposed in the literature and some of them highlighted the need of a better understanding of retention properties to lead to a rational choice of columns. However, unlike classical chromatographic methods, the problem of their robustness evaluation has often been left unaddressed. In the present study, we present a robustness study that was applied to the chromatographic testing procedure we had developed and optimized previously. A design of experiment (DoE) approach was implemented. Four factors, previously identified as potentially influent, were selected and subjected to small controlled variations: solvent fraction, temperature, pH and buffer concentration. As our model comprised quadratic terms instead of a simple linear model, we chose a D-optimal design in order to minimize the experiment number. As a previous batch-to-batch study [K. Le Mapihan, Caractérisation et classification des phases stationnaires utilisées pour l'analyse CPL de produits pharmaceutiques, Ph.D. Thesis, Pierre and Marie Curie University, 2004] had shown a low variability on the selected stationary phase, it was then possible to split the design into two parts, according to the solvent nature, each using one column. Actually, our testing procedure involving assays both with methanol and with acetonitrile as organic modifier, such an approach enabled to avoid a possible bias due to the column ageing considering the number of experiments required (16 + 6 center points). Experimental results were computed thanks to a Partial Least Squares regression procedure, more adapted than the classical regression to handle factors and responses not completely independent. The results showed the behavior of the solutes in relation to their physico-chemical properties and the relevance of the second term degree of our model. Finally, the robust domain of the test has been fairly identified, so that any potential user precisely knows to which extend each experimental parameter must be controlled when our testing procedure is to be implemented.