Nile Red Quantifier:A novel and quantitative tool to study lipid accumulation in patient-derived circulating monocytes using confocal microscopy

The inflammatory profile of circulating monocytes is an important biomarker for atherosclerotic plaque vulnerability. Recent research revealed that peripheral lipid uptake by monocytes alters their phenotype toward an inflammatory state and this coincides with an increased lipid droplet (LD) content. Determination of lipid content of circulating monocytes is, however, not very well established. Based on Nile Red (NR) neutral LD imaging, using confocal microscopy and computational analysis, we developed NR Quantifier (NRQ), a novel quantification method to assess LD content in monocytes. Circulating monocytes were isolated from blood and used for the NR staining procedure. In monocytes stained with NR, we clearly distinguished, based on 3D imaging, phospholipids and exclusively intracellular neutral lipids. Next, we developed and validated NRQ, a semi-automated quantification program that detects alterations in lipid accumulation. NRQ was able to detect LD alterations after ex vivo exposure of isolated monocytes to freshly isolated LDL in a time-and dose-dependent fashion. Finally, we validated NRQ in patients with familial hypercholesterolemia and obese subjects in pre- and postprandial state. In conclusion, NRQ is a suitable tool to detect even small differences in neutral LD content in circulating monocytes using NR staining

Physical characteristics of the back are not predictive of low back pain in healthy workers: A prospective study

Background. In the working population, back disorders are an important reason for sick leave and permanent work inability. In the context of fitting the job to the worker, one of the primary tasks of the occupational health physician is to evaluate the balance between work-related and individual variables. Since this evaluation of work capacity often consists of a physical examination of the back, the objective of this study was to investigate whether a physical examination of the low back, which is routinely performed in occupational medicine, predicts the development of low back pain (LBP). Methods. This study is part of the Belgian Low Back Cohort (BelCoBack) Study, a prospective study to identify risk factors for the development of low back disorders in occupational settings. The study population for this paper were 692 young healthcare or distribution workers (mean age of 26 years) with no or limited back antecedents in the year before inclusion. At baseline, these workers underwent a standardised physical examination of the low back. One year later, they completed a questionnaire on the occurrence of LBP and some of its characteristics. To study the respective role of predictors at baseline on the occurrence of LBP, we opted for Cox regression with a constant risk period. Analyses were performed separately for workers without any back antecedents in the year before inclusion ('asymptomatic' workers) and for workers with limited back antecedents in the year before inclusion ('mildly symptomatic' workers). Results. In the group of 'asymptomatic' workers, obese workers showed a more than twofold-increased risk on the development of LBP as compared to non-obese colleagues (RR 2.57, 95%CI: 1.09 - 6.09). In the group of 'mildly symptomatic' workers, the self-reports of pain before the examination turned out to be most predictive (RR 3.89, 95%CI: 1.20 - 12.64). Conclusion. This study showed that, in a population of young workers wh no or limited antecedents of LBP at baseline, physical examinations, as routinely assessed in occupational medicine, are not useful to predict workers at risk for the development of back disorders one year later