In vitro effects of photobiomodulation therapy on 50B11 sensory neurons: evaluation of cell metabolism,oxidative stress,mitochondrial membrane potential (MMP), and capsaicin-induced calcium flow

The analgesic properties of photobiomodulation therapy (PBMT) have been raising increasing interest in the clinical community due to the positive effects observed on patients, nevertheless the mechanistic basis of its action on peripheral sensory neurons remains still elusive. In this study, the effect of near-infrared (NIR) PBMT at 800 and 970 nm of wavelength was investigated on the 50B11 immortalized nociceptive sensory neuronal cell line by evaluating capsaicin-induced calcium flow and different markers correlated to mitochondria, that is, ATP, reactive oxygen species (ROS), and mitochondrial membrane potential (MMP). Calcium peak stimulated by capsaicin, the ligand of TRPV1 channel, was decreased in neurons pre-irradiated with the combination of the two wavelengths. Furthermore, delivering the 800 and 970 nm separately an increment of ATP, as well as MMP hyperpolarization were detected; notably, the 800 nm wavelength also increased ROS and O₂⁻ levels. Our findings, obtained on an in vitro model of nociception, show the positive effect of PBMT on two potential photo-targets of NIR light, namely the TRPV1 channel and the mitochondria.     

Colorimetric method for the assay of heparin content in immobilized heparin preparations

The properties of the metachromatic dye toluidine blue have been utilized to determine colorimetrically the amount of heparin covalently coupled to Sepharose. The method involves monitoring the dye depletion in the supernatant at 631 nm as Toluidine blue is adsorbed onto the heparin polymer upon the beaded matrix. The procedure represents a simple assay technique which allows the direct quantitation of heparin in immobilized heparin preparations.     

Quantitation of proteins using a dye-metal-based colorimetric protein assay

We describe a dye-metal (polyhydroxybenzenesulfonephthalein-type dye and a transition metal) complex-based total protein determination method. The binding of the complex to protein causes a shift in the absorption maximum of the dye-metal complex from 450 to 660 nm. The dye-metal complex has a reddish brown color that changes to green on binding to protein. The color produced from this reaction is stable and increases in a proportional manner over a broad range of protein concentrations. The new Pierce 660 nm Protein Assay is very reproducible, rapid, and more linear compared with the Coomassie dye-based Bradford assay. The assay reagent is room temperature stable, and the assay is a simple and convenient mix-and-read format. The assay has a moderate protein-to-protein variation and is compatible with most detergents, reducing agents, and other commonly used reagents. This is an added advantage for researchers needing to determine protein concentrations in samples containing both detergents and reducing agents.     

Diet of the Apennine hare in a southern Italy Regional Park

In this study, we examined the annual diet composition of Lepus corsicanus in two different sites within a southern Italy Regional Park. Vegetation of site 1 was composed of a mixed scrub forest (Viburno–Quercetum ilicis s.l.), a ripisilva (Roso sempervirentis–Populetum nigrae), some thermophilous scrubs (Pruno–Rhamnetalia alaterni), and a Pinus halepensis reforested area. Site 2 comprised a mixed-oak forest (Centaureo–Quercetum pubescentis s.l.) with meadows and arable lands. Micro-histological analysis of faecal samples revealed that hares utilised 70 different species of plants during the year, indicating the capability of the Apennine hare of exploiting a wide variety of vegetation. Herbaceous plants (Hemicryptophytes, particularly graminoid grasses, and Geophytes) predominated in the diet. Brachypodium sylvaticum (9.44 %) and Allium subhirsutum (8.28 %) were the major contributors to the diet in sites 2 and 1, respectively. Other taxa found most often in the diet were Trifolium pratense (site 1: 8.19 %; site 2: 5.80 %) and Prunus spinosa (site 1: 7.03 %; site 2: 4.10 %). Significant differences were found between sites in terms of diet richness, diversity, and evenness. Nevertheless, both the similarity indices (Morisita–Horn: 0.79; Sørensen: 0.87) showed that the food composition of the hare’s diet was broadly the same in both sites. Some qualitative and quantitative differences between sites were due to the availability or consumption of some plant species and evidenced that the Apennine hare can modify its trophic niche in order to adapt its dietary requirements to the availability of food.