Inflammatory and Metabolic Signaling Interfaces of the Hypertrophic and Senescent Chondrocyte Phenotypes Associated with Osteoarthritis.

Osteoarthritis (OA) is a complex disease of whole joints with progressive cartilage matrix degradation and chondrocyte transformation. The inflammatory features of OA are reflected in increased synovial levels of IL-1ß, IL-6 and VEGF, higher levels of TLR-4 binding plasma proteins and increased expression of IL-15, IL-18, IL-10 and Cox2, in cartilage. Chondrocytes in OA undergo hypertrophic and senescent transition; in these states, the expression of Sox-9, Acan and Col2a1 is suppressed, whereas the expression of RunX2, HIF-2α and MMP-13 is significantly increased. NF-kB, which triggers many pro-inflammatory cytokines, works with BMP, Wnt and HIF-2α to link hypertrophy and inflammation. Altered carbohydrate metabolism and the upregulation of GLUT-1 contribute to the formation of end-glycation products that trigger inflammation via the RAGE pathway. In addition, a glycolytic shift, increased rates of oxidative phosphorylation and mitochondrial dysfunction generate reactive oxygen species with deleterious effects. An important surveyor mechanism, the YAP/TAZ signaling system, controls chondrocyte differentiation, inhibits ageing by protecting the nuclear envelope and suppressing NF-kB, MMP-13 and aggrecanases. The inflammatory microenvironment and synthesis of key matrix components are also controlled by SIRT1 and mTORc. Senescent chondrocytes represent the functional end stage of hypertrophic differentiation and characteristically upregulate p16 and p21, but also a variety of inflammatory cytokines, chemokines and metalloproteinases, developing the senescence-associated secretory phenotype. Senolysis with dendrobin, miR29b-5p and other agents has been shown to be efficient under experimental conditions, and appears to be a promising tool for the treatment of OA, as it restores COL2A1 and aggrecan synthesis, suppressing NF-kB and destructive metalloproteinases.

An optimized image analysis algorithm for detecting nuclear signals in digital whole slides for histopathology.

Nuclear estrogen receptor (ER), progesterone receptor (PR) and Ki-67 protein positive tumor cell fractions are semiquantitatively assessed in breast cancer for prognostic and predictive purposes. These biomarkers are usually revealed using immunoperoxidase methods resulting in diverse signal intensity and frequent inhomogeneity in tumor cell nuclei, which are routinely scored and interpreted by a pathologist during conventional light-microscopic examination. In the last decade digital pathology-based whole slide scanning and image analysis algorithms have shown tremendous development to support pathologists in this diagnostic process, which can directly influence patient selection for targeted- and chemotherapy. We have developed an image analysis algorithm optimized for whole slide quantification of nuclear immunostaining signals of ER, PR, and Ki-67 proteins in breast cancers. In this study, we tested the consistency and reliability of this system both in a series of brightfield and DAPI stained fluorescent samples. Our method allows the separation of overlapping cells and signals, reliable detection of vesicular nuclei and background compensation, especially in FISH stained slides. Detection accuracy and the processing speeds were validated in routinely immunostained breast cancer samples of varying reaction intensities and image qualities. Our technique supported automated nuclear signal detection with excellent efficacy: Precision Rate/Positive Predictive Value was 90.23 ± 4.29%, while Recall Rate/Sensitivity was 88.23 ± 4.84%. These factors and average counting speed of our algorithm were compared with two other open source applications (QuPath and CellProfiler) and resulted in 6-7% higher Recall Rate, while 4- to 30-fold higher processing speed. In conclusion, our image analysis algorithm can reliably detect and count nuclear signals in digital whole slides or any selected large areas i.e. hot spots, thus can support pathologists in assessing clinically important nuclear biomarkers with less intra- and interlaboratory bias inherent of empirical scoring. © 2017 International Society for Advancement of Cytometry.

Safety of anastomotic techniques and consequences of anastomotic leakage in patients with colorectal cancer: a single surgeon experience.

INTRODUCTION: Colorectal cancer is a common type of malignant disease of the digestive tract. Anastomotic leakage (AL) still represents a serious complication in gastrointestinal surgery, associated with high morbidity and mortality. METHODS: We conducted a retrospective case-control study and analyzed a single surgeon's data about 359 patients treated for colorectal cancer. Patients were divided as follows: Study Group (patients with AL - 37 patients) and Control Group (patients without AL - 322 patients). Surgical and anastomotic technique-related information was processed. RESULTS: Surgical procedures for right sided colon tumors resulted in a significantly lower rate of anastomotic leakage (P=0.0231). For left sided colectomies end to end handsewn double layer anastomosis presented decreased odds (OR=0.176). For sigmoid segmental resection end to end anastomotic techniques developed low rate of fistula formation (handsewn - OR=0.593, stapled - OR=0.685). Performing Dixon type surgical interventions, anastomotic techniques seemed without influence on anastomotic leak appearance (handsewn and stapled), although distal anastomoses were identified as significant risk factors for fistula formation (P=0.0017). In order to perform subtotal colectomy, side to side sutures (handsewn and stapled) seemed safe choices for anastomotic procedure (P=0.0073). Patient with anastomotic leakage suffered a significantly longer hospital stay (P=0.0079), presented higher rate of surgical reintervention (P=0.0001), increased mortality (P=0.0001) and elevated hospitalization costs (P=0.0079). CONCLUSION: Postoperative complications like anastomosis leakage significantly increase hospitalization period, necessity of surgical reintervention, mortality and financial costs. In order to avoid these unpleasant events, bowel anastomoses require standardization during surgery.