The Different Phytochemical Profiles of Salvia officinalis Dietary Supplements Labelled for Menopause Symptoms.

Phytochemical screening of four commercial products containing Salvia officinalis was carried out. Total phenolic content was estimated spectrophotometrically through the use of the Folin-Ciocalteau method, flavonoid content was measured through the use of aluminum chloride and 2,4-dinitrophenylhydrazine colorimetric assays, and isoflavones and α/ß-thujones were analyzed through the use of high-performance liquid chromatograph (HPLC) and the gas chromatographic method. The analyses revealed the absence of thujones and isoflavones (i.e., genistin, genistein, and daidzein) in all four different extracts. The content of polyphenolic compounds varied among the samples, with the extract T being richer in both polyphenols and flavonoids than the other products by 1.8-3.2 and 1.4-4.0 times, respectively (p-value < 0.05). These results highlight the importance of quality control in salvia-based products since a thujone-free extract rich in polyphenols and flavonoids could be a good candidate for further preclinical and clinical studies to identify an effective herbal approach suitable for the long-term therapy of menopausal symptoms.

The atypical receptor CCRL2 is required for CXCR2-dependent neutrophil recruitment and tissue damage.

CCRL2 is a 7-transmembrane domain receptor that shares structural and functional similarities with the family of atypical chemokine receptors (ACKRs). CCRL2 is upregulated by inflammatory signals and, unlike other ACKRs, it is not a chemoattractant-scavenging receptor, does not activate ß-arrestins, and is widely expressed by many leukocyte subsets. Therefore, the biological role of CCRL2 in immunity is still unclear. We report that CCRL2-deficient mice have a defect in neutrophil recruitment and are protected in 2 models of inflammatory arthritis. In vitro, CCRL2 was found to constitutively form homodimers and heterodimers with CXCR2, a main neutrophil chemotactic receptor. By heterodimerization, CCRL2 could regulate membrane expression and promote CXCR2 functions, including the activation of ß2-integrins. Therefore, upregulation of CCRL2 observed under inflammatory conditions is functional to finely tune CXCR2-mediated neutrophil recruitment at sites of inflammation.

An innovative cell-based assay for the detection of modulators of soluble guanylate cyclase.

Guanylate cyclase (GC) catalyzes the biosynthesis of cyclic guanosine 3',5'- monophosphate (cGMP) from GTP. GC exists in two isoenzyme forms: soluble and membrane-bound. The soluble GC (sGC) is a heterodimer composed of an alpha and a beta subunit, and it contains heme as a prosthetic group. The most important physiological activator of sGC is nitric oxide, which activates the enzyme upon binding to the heme moiety. By producing the second messenger cGMP, which regulates various effector systems such as phosphodiesterases, ion channels, and protein kinases, sGC plays an important role in different physiological processes, thus representing a very attractive pharmacological target. In fact, the pathogenesis of several diseases, especially those of the cardiovascular system, has been linked to inappropriate regulation of sGC. In order to find new modulators for this important enzyme, an innovative cell-based assay has been developed and optimized for the use in high-throughput screening. This luminescent assay, which is suitable for both 96- and 384-well plate formats, has been achieved by stably expressing the alpha and beta subunits of a mutated form of sGC in Chinese hamster ovary cells. The mutated form synthesizes cyclic adenosine 3',5'-monophosphate instead of cGMP, allowing the detection of enzymatic activity by a reporter gene approach. We demonstrated that this cell line responds to compounds typically used in the field of sGC research and that it represents an innovative and robust assay to screen for sGC modulators with high efficiency and high sensitivity by means of standard luminescence readers.