Search

Context The search for somatic mutations in adrenals resected from patients with primary aldosteronism (PA) is performed by Sanger sequencing, often implemented with immunohistochemistry (IHC)-guidance focused on aldosterone-producing (CYP11B2-positive) areas. Objective To investigate the impact of double IHC for CYP11B1 and CYP11B2 on Sanger and next-generation sequencing (NGS). Methods We investigated 127 consecutive adrenal aldosterone-producing adenomas from consenting surgically cured PA patients using double IHC for CYP11B1 and CYP11B2, by Sanger sequencing and NGS. Results Double IHC for CYP11B2 and CYP11B1 revealed 3 distinct patterns: CYP11B2-positive adenoma (pattern 1), mixed CYP11B1/CYP11B2-positive adenoma (pattern 2), and adrenals with multiple small CYP11B2-positive nodules (pattern 3). Sanger sequencing allowed detection of KCNJ5 mutations in 44% of the adrenals; NGS revealed such mutations in 10% of those negative at Sanger and additional mutations in 61% of the cases. Importantly the rate of KCNJ5 mutations differed across patterns: 17.8% in pattern 1, 71.4% in pattern 2, and 10.7% in pattern 3 (χ2 = 22.492, P <.001). Conclusion NGS allowed detection of mutations in many adrenals that tested negative at Sanger sequencing. Moreover, the different distribution of KCNJ5 mutations across IHC patterns indicates that IHC-guided sequencing protocols selecting CYP11B2-positive areas could furnish results that might not be representative of the entire mutational status of the excised adrenal, which is important at a time when KCNJ5 mutations are suggested to drive management of patients with aldosterone-producing adenomas. [ABSTRACT FROM AUTHOR]

Background: The aim of this study was to retrospectively evaluate the 3-year radiographic outcomes of periodontal intrabony defects treated with non-surgical subgingival therapy (NST), assessing radiographic bone gain (RBG) through experimental digital software, named "Bone Defect Analysis (BDA)". Methods: The study included 17 intrabony defects in 14 patients. BDA software (version 1) was used on radiographs to calculate RBG (in %) and variations in defect angle (in °) between baseline (T0) and 3-year follow-up (T1). Soft tissue conditions were registered, reporting bleeding on probing (BOP), probing pocket depth (PPD), and clinical attachment level (CAL). Defects were analyzed according to angles less (group A) or greater (group B) than 30°. Results: Nine and eight defects were, respectively, analyzed in groups A and B. Three years after treatment, an average RBG of 12.28% was found overall, with 13.25% and 10.11% for groups A and B, respectively (p = 0.28). Clinically, a mean CAL of 6.05 mm at T1 (from 10.94 mm at T0) was found, with 6.88 mm and 5.12 mm in groups A and B, respectively (p = 0.07). Conclusions: BDA software demonstrated predictability in the evaluation of bone variations after NST, revealing better clinical findings for intrabony defects with an initial smaller angle. [ABSTRACT FROM AUTHOR]

Background: Osteoarthritis (OA) is a disabling disease affecting all joint tissues and particularly articular cartilage. Moreover, inflammatory process and mediators plays an important role in the initiation and progression of OA determining cartilage extracellular matrix degradation. Aim: The aim of this study is to evaluate the effects of β-caryophyllene (BCP), a plantderived sesquiterpene with anti-inflammatory and antioxidant properties, alone or in combination with ascorbic acid (AA) and d-glucosamine (GlcN) on human primary chondrocytes inflamed with macrophage-derived conditioned medium (CM). Materials and Methods: Cartilage biopsies were obtained from OA patients and controls. Histological staining and immunohistochemistry for inflammatory markers were performed to characterize the tissues. Chondrocytes cell viability, gene expression of pro-inflammatory mediators, metalloproteinases (MMPs), collagen type II and aggrecan levels were analyzed in primary human chondrocytes isolated from OA patients and exposed to the CM of activated U937 monocytes and subsequently treated with BCP alone or in combination with AA and GlcN. Results: Histology and immunohistochemistry analysis pointed out cartilage degeneration, fragmentation and fissuring in OA patients, associated with the presence of positivity to inflammatory molecules compared to controls. The macrophage-derived CM induced a decrease of chondrocyte viability and a significantly increase of inflammatory molecules and MMPs expression. When BCP was added to the inflamed cells, alone or in combination with AA and GlcN, there was an increase of cell viability, a decrease of gene expression of interleukin-1 beta, NF-κB1 and MMP-13 and an increase of ECM matrix molecules (aggrecan and type II collagen). Conclusions: Low-doses of BCP are effective in counteracting the inflammatory effects determined by macrophage-derived CM stimulation on chondrocytes. Moreover, the combined used of BCP, AA and GlcN seems to have synergistic effects in protecting chondrocytes. [ABSTRACT FROM AUTHOR]