Sensitivity to Cisplatin in Head and Neck Cancer Cells Is Significantly Affected by Patient-Derived Cancer-Associated Fibroblasts

Cancer-associated fibroblasts (CAFs) are one of the most abundant and critical components of the tumor stroma. CAFs can impact many important steps of cancerogenesis and may also influence treatment resistance. Some of these effects need the direct contact of CAFs and cancer cells, while some involve paracrine signals. In this study, we investigated the ability of head and neck squamous cell carcinomas (HNSCC) patient-derived CAFs to promote or inhibit the colony-forming ability of HNSCC cells. The effect of cisplatin on this promoting or inhibiting influence was also studied. The subsequent analysis focused on changes in the expression of genes associated with cancer progression. We found that cisplatin response in model HNSCC cancer cells was modified by coculture with CAFs, was CAF-specific, and different patient-derived CAFs had a different “sensitizing ratio”. Increased expression of VEGFA, PGE2S, COX2, EGFR, and NANOG in cancer cells was characteristic for the increase of resistance. On the other hand, CCL2 expression was associated with sensitizing effect. Significantly higher amounts of cisplatin were found in CAFs derived from patients who subsequently experienced a recurrence. In conclusion, our results showed that CAFs could promote and/or inhibit colony-forming capability and cisplatin resistance in HNSCC cells via paracrine effects and subsequent changes in gene expression of cancer-associated genes in cancer cells.     

The Attenuated Secretion of Hyaluronan by UVA-Exposed Human Fibroblasts Is Associated with Up- and Downregulation of HYBID and HAS2 Expression via Activated and Inactivated Signaling of the p38/ATF2 and JAK2/STAT3 Cascades

Little is known about the effects on hyaluronan (HA) metabolism of UVA radiation. This study demonstrates that the secretion of HA by human dermal fibroblasts (HDFs) is downregulated by UVA, accompanied by the down- and upregulation of mRNA and protein levels of the HA-synthesizing enzyme (HAS2) and the HA-degrading protein, HYaluronan Binding protein Involved in HA Depolymerization(HYBID), respectively. Signaling analysis revealed that the exposure distinctly elicits activation of the p38/MSK1/CREB/c-Fos/AP-1 axis, the JNK/c-Jun axis, and the p38/ATF-2 axis, but downregulates the phosphorylation of NF-kB and JAK/STAT3. A signal inhibition study demonstrated that the inhibition of p38 significantly abrogates the UVA-accentuated mRNA level of HYBID. Furthermore, the inhibition of STAT3 significantly downregulates the level of HAS2 mRNA in non-UVA exposed HDFs. Analysis using siRNAs demonstrated that transfection of ATF-2 siRNA but not c-Fos siRNA abrogates the increased protein level of HYBID in UVA-exposed HDFs. An inhibitor of protein tyrosine phosphatase but not of protein serine/threonine phosphatase restored the diminished phosphorylation level of STAT3 at Tyr 705, accompanied by a significant abolishing effect on the decreased mRNA expression level of HAS2. Silencing with a protein tyrosine phosphatase PTP-Meg2 siRNA revealed that it abrogates the decreased phosphorylation of STAT3 at Tyr 705 in UVA-exposed HDFs. These findings suggest that the UVA-induced decrease in HA secretion by HDFs is attributable to the down- and upregulation of HAS2 and HYBID expression, respectively, changes that are mainly ascribed to the inactivated signaling of the STAT3 axis due to the activated tyrosine protein phosphatase PTP-Meg2 and the activated signaling of the p38/ATF2 axis, respectively.     

The Co-Culture of Staphylococcal Biofilm and Fibroblast Cell Line: The Correlation of Biological Phenomena with Metabolic NMR1 Footprint

Staphylococcus aureus is one of the most prevalent pathogens associated with several types of biofilm-based infections, including infections of chronic wounds. Mature staphylococcal biofilm is extremely hard to eradicate from a wound and displays a high tendency to induce recurring infections. Therefore, in the present study, we aimed to investigate in vitro the interaction between S. aureus biofilm and fibroblast cells searching for metabolites that could be considered as potential biomarkers of critical colonization and infection. Utilizing advanced microscopy and microbiological methods to examine biofilm formation and the staphylococcal infection process, we were able to distinguish 4 phases of biofilm development. The analysis of staphylococcal biofilm influence on the viability of fibroblasts allowed us to pinpoint the moment of critical colonization—12 h post contamination. Based on the obtained model we performed a metabolomics analysis by ¹H NMR spectroscopy to provide new insights into the pathophysiology of infection. We identified a set of metabolites related to the switch to anaerobic metabolism that was characteristic for staphylococcal biofilm co-cultured with fibroblast cells. The data presented in this study may be thus considered a noteworthy but preliminary step in the direction of developing a new, NMR-based tool for rapid diagnosing of infection in a chronic wound.     

Classification of cardiac sound signals using constrained tunable-Q wavelet transform

The features extracted from the cardiac sound signals are commonly used for detection and identification of heart valve disorders. In this paper, we present a new method for classification of cardiac sound signals using constrained tunable-Q wavelet transform (TQWT). The proposed method begins with a constrained TQWT based segmentation of cardiac sound signals into heart beat cycles. The features obtained from heart beat cycles of separately reconstructed heart sounds and murmur can better represent the various types of cardiac sound signals than that from containing both. Therefore, heart sounds and murmur have been separated using constrained TQWT. Then the proposed novel raw feature set has been created by the parameters that have been optimized while constraining the output of TQWT together with that of extracted by using time-domain representation and Fourier–Bessel (FB) expansion of separated heart sounds and murmur. However, the adaptively selected features have been used to obtain the final feature set for subsequent classification of cardiac sound signals using least squares support vector machine (LS-SVM) with various kernel functions. The performance of the proposed method has been validated with publicly available datasets and the results have been compared with the existing short-time Fourier transform (STFT) based method. The proposed method shows higher percentage classification accuracy of 94.01 as compared to 93.53 of STFT based method. In comparison with STFT based method, it is noteworthy that the proposed method uses well defined and lower dimensionality of feature vector that can reduce the computational complexity.     

Segmentation of the left ventricle in cardiac cine MRI using a shape-constrained snake model

Segmentation of the left ventricle (LV) is a hot topic in cardiac magnetic resonance (MR) images analysis. In this paper, we present an automatic LV myocardial boundary segmentation method using the parametric active contour model (or snake model). By convolving the gradient map of an image, a fast external force named gradient vector convolution (GVC) is presented for the snake model. A circle-based energy is incorporated into the GVC snake model to extract the endocardium. With this prior constraint, the snake contour can conquer the unexpected local minimum stemming from artifacts and papillary muscle, etc. After the endocardium is detected, the original edge map around and within the endocardium is directly set to zero. This modified edge map is used to generate a new GVC force filed, which automatically pushes the snake contour directly to the epicardium by employing the endocardium result as initialization. Meanwhile, a novel shape-similarity based energy is proposed to prevent the snake contour from being strapped in faulty edges and to preserve weak boundaries. Both qualitative and quantitative evaluations on our dataset and the publicly available database (e.g. MICCAI 2009) demonstrate the good performance of our algorithm.     

Quantification of heart fatty acid binding protein as a biomarker for drug-induced cardiac and musculoskeletal necroses

Heart fatty acid binding protein (Fabp3) is a cytosolic protein expressed primarily in heart, and to a lesser extent in skeletal muscle, brain, and kidney. During myocardial injury, the Fabp3 level in serum is elevated rapidly, making it an ideal early marker for myocardial infarction. In this study, an MS‐based selected reaction monitoring method (LC‐SRM) was developed for quantifying Fabp3 in rat serum. Fabp3 was enriched first through an immobilized antibody, and the protein was digested on beads directly. A marker peptide of Fabp3 was quantified using LC‐SRM with a stable isotope‐labeled peptide standard. For six quality control samples with Fabp3 ranging from 0.256 to 25 ng, the average recovery following the procedure was about 73%, and the precision (%CV) between replicates was less than 7%. The Fabp3 concentrations in rat serum peaked 1 h after isoproterenol treatment, and returned to baseline levels 24 h after the dose. Elevated Fabp3 levels were also detected in rats administered with a PPAR α/δ agonist, which has shown to cause skeletal muscle necrosis. Fabp3 can be used as a biomarker for both cardiac and skeletal necroses. The cross‐validation of the LC‐SRM method with an existing ELISA method is described.     

Metabolic and cardiovascular responses to exercise with caffeine

The effect of caffeine on circulation and metabolism was studied during a discontinuous maximal oxygen uptake (VO₂ max) test on a bicycle ergometer. Eight male subjects were chosen for their minimal use of caffeine of which six normally did not ingest caffeinated beverages. Two caffeinated and two decaffeinated (control) trials were administered to each subject following double-blind and counterbalanced testing procedures. One hour before exercise, subjects ingested decaffeinated coffee with or without 350 mg of caffeine anhydrous added. Cardiac output (Q) was measured by a C0₂ rebreathing technique. Compared to control trials, caffeine increased the VO₂ max by an average of 140ml min?¹ (p <0-0·5)l. In addition, during caffeine trials, the average maximal heart rate (HR max) was elevated by 5 beats min^?1 (p<0·01). There were no other statistically significant changes in metabolic (VO₂ and R) and cardiovascular (Q, HR, SV and (a-v) 0₂ diff) variables during either submaximal or maximal exercise. Performance, as measured by total exercise time on the VO₂ max test, was not significantly different (p>0·05). It was concluded that caffeine has a small physiological but a statistically significant effect on VQ₂ max and HR max. During submaximal exercise, caffeine did not significantly effect the cardiovascular system.