Calibrating single-ended fiber-optic Raman spectra distributed temperature sensing data.

Hydrologic research is a very demanding application of fiber-optic distributed temperature sensing (DTS) in terms of precision, accuracy and calibration. The physics behind the most frequently used DTS instruments are considered as they apply to four calibration methods for single-ended DTS installations. The new methods presented are more accurate than the instrument-calibrated data, achieving accuracies on the order of tenths of a degree root mean square error (RMSE) and mean bias. Effects of localized non-uniformities that violate the assumptions of single-ended calibration data are explored and quantified. Experimental design considerations such as selection of integration times or selection of the length of the reference sections are discussed, and the impacts of these considerations on calibrated temperatures are explored in two case studies.

Chlamydia trachomatis Infection Leads to Defined Alterations to the Lipid Droplet Proteome in Epithelial Cells.

The obligate intracellular bacterium Chlamydia trachomatis is a major human pathogen and a main cause of genital and ocular diseases. During its intracellular cycle, C. trachomatis replicates inside a membrane-bound vacuole termed an "inclusion". Acquisition of lipids (and other nutrients) from the host cell is a critical step in chlamydial replication. Lipid droplets (LD) are ubiquitous, ER-derived neutral lipid-rich storage organelles surrounded by a phospholipids monolayer and associated proteins. Previous studies have shown that LDs accumulate at the periphery of, and eventually translocate into, the chlamydial inclusion. These observations point out to Chlamydia-mediated manipulation of LDs in infected cells, which may impact the function and thereby the protein composition of these organelles. By means of a label-free quantitative mass spectrometry approach we found that the LD proteome is modified in the context of C. trachomatis infection. We determined that LDs isolated from C. trachomatis-infected cells were enriched in proteins related to lipid metabolism, biosynthesis and LD-specific functions. Interestingly, consistent with the observation that LDs intimately associate with the inclusion, a subset of inclusion membrane proteins co-purified with LD protein extracts. Finally, genetic ablation of LDs negatively affected generation of C. trachomatis infectious progeny, consistent with a role for LD biogenesis in optimal chlamydial growth.