Search

Forested wetlands dominated by baldcypress ( Taxodium distichum) and water tupelo (Nyssa aquatica ) are commonly found in coastal regions of the southeastern United States. Global climate change and in particular sea level rise will alter the frequency and magnitude of wet/dry periods and salinity levels in these ecosystems. Soil microcosm experiments were set up to identify the effects of water level variations (0.4–3.0 g-water g-soil −1 ) and salinity changes (0, 1 and 5 ppt of NaCl) on greenhouse gas emissions (CH 4 , CO 2 , and N 2 O) and dissolved organic carbon (DOC) characteristics from forested wetland soils. Our results indicate that, the effect of water level was much greater than salt intrusion on C and N cycling. Wet–dry treatments significantly decreased DOC production and total CH 4 -C loss, aromatic and humic-like substance compounds in DOC were increased in both flooding and wet–dry treatments after 60-d incubation. The molecular weight (MW) of DOC, as indicated by E2/E3 ratio and spectral slope, after flooding treatments was higher than that in wet–dry treatments. A first order kinetic model showed there was a positive linear correlation (r 2 = 0.73) between CO 2 emission rate and DOC concentration which indicated that CO 2 was mainly generated from DOC. An exponential kinetic model was applied to describe the correlation between CH 4 emission rate and DOC concentration (r 2 = 0.41). This study demonstrates that an increase in salinity, and in particular variations in wet–dry cycles, will lead to changes in the formation of climate-relevant greenhouse gases, such as CH 4 , CO 2 , and N 2 O. [ABSTRACT FROM AUTHOR]

Little is known about the potential activity of microbial communities in hypersaline sediment ecosystems. Ribosomal tag libraries of DNA and RNA extracted from the sediment of Lake Strawbridge (Western Australia) revealed bacterial and archaeal operational taxonomic units (OTUs) with high RNA/DNA ratios providing evidence for the presence of ‘rare’ but potentially “active” taxa. Among the ‘rare’ bacterial taxaHalomonas, SalinivibrioandIdiomarinashowed the highest protein synthesis potential. Rare but ‘active’ archaeal OTUs were related to the KTK 4A cluster and the Marine-Benthic-Groups B and D. We present the first molecular analysis of the microbial diversity and protein synthesis potential of rare microbial taxa in a hypersaline sediment ecosystem. [ABSTRACT FROM PUBLISHER]

Abstract: We determined circulating T helper, T regulatory and ICOS+T regulatory as well as DC cell counts in 29 patients with cerebral gliomas. Samples from patients with gliomas vs. healthy controls and from patients with glioblastomas vs. patients with glioma WHO grades I–III contained significantly (p<0.05) decreased numbers of total as well as mature, i.e. myeloid and plasmacytoid DCs. Patients with glioblastomas demonstrated significantly lower values of CD4+ as well as an increased fraction of ICOS+T regulatory/CD4+ cells. Higher CD4+ cell counts (≥225cells/μl, median) were associated with improved survival in glioblastomas. [Copyright &y& Elsevier]

Torrential rain and extreme flooding caused by Atlantic hurricanes mobilize a large pool of organic matter (OM) from coastal forested watersheds in the southeastern United States. However, the mobilization of toxic metals such as mercury (Hg) that are associated with this vast pool of OM are rarely measured. This study aims to assess the variations of total Hg (THg) and methylmercury (MeHg) levels and the isotopic compositions of Hg in a blackwater river (Waccamaw River, SC, U.S.A.) during two recent extreme flooding events induced by Hurricane Joaquin (October 2015) and Hurricane Matthew (October 2016). We show that extreme flooding considerably increased filtered THg and MeHg concentrations associated with aromatic dissolved organic matter. During a 2‐month sampling window each year (October–November), we estimate that about 27% (2015) and 78% (2016) of the average amount of Hg deposited atmospherically during these 2 months was exported via the river. The isotopic composition of Hg in the river waters was changed only slightly by the substantial inputs of runoff from surrounding landscapes, in which mass‐dependent fractionation (as δ202Hg) decreased from −1.47 to −1.67‰ and mass‐independent fractionation (as ∆199Hg) decreased from −0.15 to −0.37‰. The slight variations in Hg isotopic composition during such extreme flooding events imply that sources of Hg in the river are nearly unchanged even under the very high wet deposition of Hg derived from the intensive rainfall. The majority of Hg exported by the river (74–85%) is estimated to have been derived from dry deposition to the watersheds. An increase in frequency and intensity of Atlantic hurricanes is expected in the next few decades due to further warming of ocean surface waters. We predict that increased hurricanes will mobilize more dry‐deposited Hg and in situ produced MeHg from these coastal watersheds where MeHg can be extensively bioaccumulated and biomagnified in the downstream aquatic food webs. [ABSTRACT FROM AUTHOR]

As direct real-time analysis techniques, selective ion flow tube mass spectrometry (SIFT-MS) and proton-transfer reaction mass spectrometry (PTR-MS) provide online measurement of volatile organic compounds (VOCs). Both techniques are widely used across several disciplines, e.g., atmospheric chemistry, food science, and medicine. However, the humidity of the sampled air greatly influences the quantified mixing ratio and must be accounted for. Here we present several improvements to a Voice200ultra SIFT-MS instrument to reduce background levels and enhance sensitivity. Increasing the sample gas flow to 125 sccm enables limits of detection (LODs) at the sub-parts-per-billion (sub-ppb) level, and the resulting humidity dependence is overcome by calibrating for humidity as well. A comparison with a PTR-QMS 500 showed detection limits of the PTR-MS still being an order of magnitude lower, whereas sensitivity was higher for SIFT-MS, and its calibration was still more robust against humidity. Thus, SIFT-MS is a suitable, lower-cost, and easy-to-use alternative for atmospheric trace gas measurements of more complex mixtures, even with isomers, at a varying humidity range. [ABSTRACT FROM AUTHOR]

As direct real-time analysis techniques, Selective Ion Flow Tube Mass Spectrometry (SIFT-MS) and Proton-Transfer Reaction Mass Spectrometry (PTR-MS) provide on-line measurement of volatile organic compounds (VOCs). Both techniques are widely-used across several disciplines, e.g. atmospheric chemistry, food science and medicine. However, the humidity of the sampled air greatly influences the quantified mixing ratio, and must be accounted for in the calibration of both platforms. Here we present both an evaluation of calibration functions taking account for humidity and a side-by-side comparison of the performance of the two instruments based upon a calibration of 15 different VOCs over a relative humidity range from 0-90% at 25 °C. While we made several improvements to a Voice 200 ultra SIFT-MS instrument to reduce background levels, overall detection limits for a PTR-QMS 500 are an order of magnitude lower. Sensitivity (here defined as the slope of the calibration curve) was higher for the SIFT-MS, and its calibration was more robust against humidity compared to PTR-MS. Thus, PTR-MS is the method of choice for simple, low-concentration, low-humidity analyses with a limited number of compounds whereas SIFT-MS is better-suited for more complex systems with varying humidity like our experiments investigating the change in VOC emissions of soils during dryout. [ABSTRACT FROM AUTHOR]

The enrichment culture KS is one of the few existing autotrophic, nitrate-reducing, Fe(II)-oxidizing cultures that can be continuously transferred without an organic carbon source. We used a combination of catalyzed amplification reporter deposition fluorescence in situ hybridization (CARD-FISH) and nanoscale secondary ion mass spectrometry (NanoSIMS) to analyze community dynamics, singlecell activities, and interactions among the two most abundant microbial community members (i.e., Gallionellaceae sp. and Bradyrhizobium spp.) under autotrophic and heterotrophic growth conditions. CARD-FISH cell counts showed the dominance of the Fe(II) oxidizer Gallionellaceae sp. under autotrophic conditions as well as of Bradyrhizobium spp. under heterotrophic conditions. We used NanoSIMS to monitor the fate of 13C-labeled bicarbonate and acetate as well as 15N-labeled ammonium at the single-cell level for both taxa. Under autotrophic conditions, only the Gallionellaceae sp. was actively incorporating 13C-labeled bicarbonate and 15N-labeled ammonium. Interestingly, both Bradyrhizobium spp. and Gallionellaceae sp. became enriched in [13C]acetate and [15N]ammonium under heterotrophic conditions. Our experiments demonstrated that Gallionellaceae sp. was capable of assimilating [13C]acetate while Bradyrhizobium spp. were not able to fix CO2, although a metagenomics survey of culture KS recently revealed that Gallionellaceae sp. lacks genes for acetate uptake and that the Bradyrhizobium sp. carries the genetic potential to fix CO2. The study furthermore extends our understanding of the microbial reactions that interlink the nitrogen and Fe cycles in the environment. [ABSTRACT FROM AUTHOR]

Two intense rainfalls [Hurricane Joaquin (2015) and Hurricane Matthew (2016)], one year apart, provided a unique opportunity to examine changes in dissolved organic matter (DOM) dynamics in coastal blackwater rivers under extreme flooding conditions in the southeastern United States. Two sites along Waccamaw River (a coastal blackwater river) and the outflow of 18 sub-basins of Yadkin-Pee Dee Basin were sampled during and after the flooding events. The peaks of dissolved organic carbon (DOC) and nitrogen (DON) concentrations were observed 18 and 23 days after peak discharge in 2015 and 2016, respectively. Moreover, DOM aromaticity and abundance of humic substances significantly increased during the same period. Separation of discharge hydrograph into surface runoff and subsurface flow suggested that temporal changes were mainly due to contributions from subsurface flow flushing organic matter from wetlands and organic-rich riparian zones. The spatial analysis highlighted the key role of the forested wetlands as the only land use that significantly correlated with both DOM quantity (DOC and DON load) and DOM composition (i.e., aromaticity). The Yadkin-Pee Dee River basin alone exported more than 474 million kg DOC into the ocean during high-flow conditions from the 2016 event, indicating that such extreme short-term events mobilized enormous amounts of organic carbon and nitrogen to the ocean. Considering the predicted increase in frequency and intensity of extreme rainfall events in the eastern U.S., the results of this study can shed light on changes in DOM dynamics that may occur under such conditions in the near future. [ABSTRACT FROM AUTHOR]

Purpose: Recent studies indicate that circulating microRNAs in serum/plasma are a novel class of non-invasive biomarkers with diagnostic and prognostic information. So far, circulating microRNAs have not been analyzed in patients with bladder cancer. Methods: We collected serum from patients with non-muscle invasive bladder cancer (NMIBC), muscle invasive bladder cancer (MIBC) and non-malignant urological disease. Total RNA was isolated from 400 μl of serum using the mirVana PARIS Kit; the artificial cel-miR-39 was spiked-in prior to RNA isolation to control different RNA isolation efficiencies. Quantitative real-time PCR was applied to measure the levels of 22 microRNAs upregulated in BCA tissue (miR-15a, miR-18a, miR-21, miR-93, miR-96, miR-103, miR-130b, miR-135b, miR-141, miR-182, miR-183, miR-190, miR-191, miR-200b, miR-422b, miR-425, miR-449b, miR-601, miR-639, miR-644, miR-649 and miR-1233) in the marker identification cohort (NMIBC, n = 11, MIBC, n = 10; controls, n = 10). The most promising serum microRNAs were tested in a validation cohort (NMIBC, n = 65, MIBC, n = 61; controls, n = 105). Results: The RNA recovery was similar in patients with NMIBC, MIBC and control subjects. The analysis of serum microRNA levels in the marker identification cohort indicated that serum miR-141 and miR-639 levels were increased in bladder cancer patients compared to CTRL. The analysis of these miR-141 and miR-639 in the validation cohort demonstrated that microRNA levels were similar in bladder cancer patients and control subjects. Furthermore, microRNA levels were not correlated with clinicopathological parameters (pT-stage, metastasis, grading). Conclusions: The analysis of serum miR-141 and miR-639 levels does not seem to be helpful in the diagnosis or prognosis of BCA. [ABSTRACT FROM AUTHOR]

Objectives: To identify an appropriate reference gene for the analysis of circulating micro-ribonucleic acid in patients with urological malignancies. Methods: Serum from patients with prostate cancer ( n = 24), bladder cancer ( n = 24), renal cell carcinoma ( n = 24) and control subjects ( n = 48) was spiked with cel-miR-39, and then ribonucleic acid was isolated. Quantitative real-time polymerase chain reaction was used to determine the levels of candidate reference genes ( RNU1-4, RNU6-2, SNORD43, SNORD44, SNORD48, S NORA74A, miR-let-7a-1, miR-106a). Reference gene stability was determined using the NormFinder, geNorm and comparative delta-Ct algorithm. The effect of normalization was tested with miR-21 as the target gene, as this was previously suggested to be upregulated in cancer patients' serum. Results: Recovery of cel-miR-39 (mean 11.6%, range 1-56%) was similar in control subjects and cancer patients. SNORD44 and SNORD74A levels were around the detection limit of the assay and were thus omitted. All remaining candidates showed satisfying stability; SNORD43 was the most stable reference gene using all three algorithms. A combination of two genes ( SNORD43, RNU1-4) increases the stability somewhat. The level of miR-21 was similar in cancer patients and healthy controls, irrespective of the normalization strategy. Conclusions: SNORD43 is a suitable reference gene for the analysis of circulating micro-ribonucleic acid in patients with urological malignancies. Our study questions the suitability of miR-21 as a biomarker for uro-oncological patients. [ABSTRACT FROM AUTHOR]

Purpose: To profile different tyrosine kinase (TK) expression patterns in clear cell renal carcinoma (ccRCC). Methods: We analysed mRNA expression levels of 89 receptor and non-receptor TK in corresponding cancer and normal renal tissue from 5 patients with ccRCC using the TaqMan Low-Density Array technology. In order to confirm aberrant TK expressions, a subsequent analysis of 25 ccRCC and corresponding normal renal tissues was performed, applying quantitative real-time PCR. To confirm mRNA expression levels on protein level, we studied ERBB4 and HCK using immunohistochemistry. Results: A total of 12 TK were significantly upregulated in ccRCC (ABL2, FLT1, BTK, HCK, JAK3, CSF1R, MET, JAK1, MATK, PTPRC, FYN and CSK), coherently 7 TK demonstrated a down-regulation (ERBB4, PDGFRA, NRTK3, SYK, ERBB2, FGFR3 and PTK7). These findings were validated by the utilization of RT-PCR for ABL2, FLT1 BTK, HCK, JAK3, CSF1R, MET, JAK1, MATK and vice versa for ERBB4 and PDGFRA. Immunohistochemistry revealed ERBB4 expression to be significantly lower in ccRCC in comparison to papillary RCC, chromophobe RCC, renal oncocytoma and normal renal tissue ( P < 0.001). HCK protein expression was reduced in ccRCC in contrast to papillary RCC ( P < 0.001) or oncocytoma ( P = 0.023), but similar to chromphobe RCC ( P = 0.470), sarcomatoid RCC ( P = 0.754) and normal renal tissue ( P = 0.083). Neither ERBB4 nor HCK were correlated ( P > 0.05) with clinical-pathological parameters. Conclusion: TK constitute valuable targets for pharmaceutical anti-cancer therapy. ERBB4 and HCK depict significantly lower expression levels in renal cancer tissues. [ABSTRACT FROM AUTHOR]

Abstract: Objective: Cell-free circulating mitochondrial DNA (mtDNA) has been proposed as universal diagnostic and prognostic biomarker in cancer patients. Patients and methods: Cell-free DNA was isolated from 1 ml serum from patients with bladder cancer (BCA, n = 84), renal cell carcinoma (RCC, n = 33), and prostate cancer (CaP, n = 23), and compared with healthy individuals (n = 79). Quantitative real-time PCR was used to analyze the levels of a 79 bp (mtDNA-79), and 220 bp (mtDNA-220) fragment of the mitochondrial specific 16S-RNA. The mitochondrial DNA integrity (mtDNA-integrity) was defined as ratio of mtDNA-220 to mtDNA-79 fragments. Results: In healthy controls, mtDNA-79 levels were increased in male volunteers; mtDNA-230 levels and mtDNA-integrity were correlated with age. Neither mtDNA levels nor mtDNA-integrity were correlated with age or gender in cancer patients. Circulating mtDNA-79 (median 8.75 × 106 vs. 0.43 × 106 copies/ml) and mtDNA-230 (8.11 × 106 vs. 0.27 × 106 copies/ml) levels were significantly increased in cancer patients and allowed sensitive (84%) and specific (97%) discrimination from healthy controls. mtDNA levels were unequally distributed among the different cancer entities (mtDNA-79: BCA 9.54 × 106 vs. RCC 6.69 × 106 vs. CaP 4.48 × 106 copies/ml; mtDNA-230: BCA 9.78 × 106 vs. RCC 6.74 × 106 vs. CaP 1.94 × 106 copies/ml). The mtDNA-integrity was increased in RCC and BCA patients compared to control subjects and CaP patients. Serum mtDNA-integrity was correlated with pathological stage in RCC and with tumor grade in BCA patients. Conclusion: Circulating mtDNA levels are associated with gender and age in healthy individuals, but not in cancer patients. Quantification of circulating mtDNA may help identify patients with urologic malignancies. [Copyright &y& Elsevier]

Global histone modification patterns have been shown to be a predictive factor of recurrence in various cancers. We analyzed global histone-3-lysine-27 (H3K27) methylation in prostate cancer (PCA) tissues. H3K27 mono-, di-, and tri-methylation patterns were different in nonmalignant prostate tissue, localized PCA, metastatic PCA, and castration-resistant PCA. H3K27 mono-methylation was correlated with pT-stage, capsular penetration, seminal vesicle infiltration, and Gleason score in localized PCA and may therefore indicate adverse prognosis. [ABSTRACT FROM AUTHOR]

What's known on the subject? and What does the study add? Epigenetic alterations are important during carcinogenesis, and earlier studies suggested that global histone modification levels are predictive for patients' outcome in various tumor entities. We demonstrate that H3K27me1 and H3K27me3 are markers of progression-free survival in patients with renal cell carcinoma. OBJECTIVE • To evaluate if histone H3 lysine 27 (H3K27) methylation plays a role in renal cell carcinoma (RCC) tissue and whether its expression is a predictor of cancer recurrence in RCC. MATERIALS AND METHODS • A tissue microarray (TMA) with 193 RCC specimens (comprising 142 clear-cell, 31 papillary, 10 chromophobe and 10 sarcomatoid RCC), 10 oncocytoma tissue specimens and a TMA with 30 benign renal tissue samples were stained with antibodies against H3K27-monomethyl (H3K27me1), H3K27-dimethyl (H3K27me2) and H3K27-trimethyl (H3K27me3). • Sections were scored according to staining intensity and the proportion of epithelial cells showing nuclear staining. • H3K27 methylation levels were correlated with established clinical-pathological variables (tumour-node-metastasis [TNM] stage, Fuhrman grade) and progression-free/cancer-specific survival. RESULTS • H3K27me1/-me2/-me3 staining was significantly more intense in papillary RCC then in clear-cell RCC. • H3K27me3 levels were higher in oncocytoma than in RCC. • H3K27me1/-me2/-me3 methylation levels were inversely correlated with Fuhrman grading and pT-stage. • Global H3K27me1/-me2/-me3 methylation levels were always higher in benign renal tissue than in RCC with tumour relapse (H3K27me1 P < 0.001, H3K27me2 P= 0.032, H3K27me3 P= 0.004). • Progression-free survival was shorter in patients with lower levels of H3K27me1 and H3K27me3 in the univariate analysis. The newly created H3K27me score (combining the staining levels of the single modifications) was a significant and independent predictor of RCC progression-free survival. CONCLUSION • The present study on H3K27-methylation supports the hypothesis that global histone modifications are potential markers of cancer prognosis in RCC. One reason could be that decreased H3K27 indicates transcriptional activation and therefore predicts cancer activation. [ABSTRACT FROM AUTHOR]

Study Type - Prognosis (case series) Level of Evidence 4 What's known on the subject? and What does the study add? Epigenetic alterations play an essential role during carcinogenesis. While DNA methylation has been extensively studied in bladder cancer, the relevance of histone modifications remains to be clarified. Earlier studies suggested that global histone modification levels are predictive for patients' outcome in various tumour entities (e.g. prostate, lung, breast and kidney cancer). The possibility to determine global histone modification levels easily and inexpensively using immunohistochemistry increases a potential routine use in the future. Our aim was therefore to investigate the global levels of histone H3K4 and H4K20 mono-, di- and trimethylation. For this purpose we prepared tissue microarrays with non-muscle-invasive bladder cancer (NMIBC), muscle-invasive bladder cancer (MIBC), bladder cancer metastases (METS) and normal urothelium (NU) tissue to compare global H3K4 and H4K20 methylation in these tissues, as well to assess the prognostic value of histone modifications. We show that global histone modification levels (H3K4me1, H3K4me3, H4K20me1, H4K20me2, H4K20me3) are lower in bladder cancer than in NU tissue. Furthermore, there was a decrease of histone modification levels (H3K4me1, H4K20me1, H4K20me2, H4K20me3) from NU over NMIBC and MIBCto METS. Histone modifications are correlated to advanced pathological stage in NMIBC and MIBC. Furthermore, H4K20me3 appeared to be a significant and independent prognostic predictor of bladder cancer-specific survival in patients with MIBC undergoing radical cystectomy. Our findings therefore provide a rationale for further investigation of histone modifications and their manipulation in bladder cancer. OBJECTIVE • To determine the role of global histone methylation as a prognostic parameter in patients with bladder cancer. PATIENTS AND METHODS • We used a tissue microarray with samples from patients with non-muscle-invasive bladder cancer (NMIBC; n= 161), muscle-invasive bladder cancer (MIBC, n= 127), normal urothelium (NU; n= 31) and bladder cancer metastases (METS; n= 31) to determine global histone methylation (me) levels at histone H3 lysine 4 (H3K4) and H4K20. RESULTS • Global histone modification levels (H3K4me1, H3K4me3, H4K20me1, H4K20me2, and H4K20me3) were lower in bladder cancersamples than in NU tissue • Global levels of H3K4me1, H4K20me1, H4K20me2 and H4K20me3 were decreasing from NU over NMIBC and MIBC to METS. • H4K20me1 levels were increased in patients with NMIBC with advanced pTstage and less differentiated bladder cancer. • In patients with MIBC, pTstage was negatively correlated with H3K4me1, H4K20me1 and H4K20me2 levels. • H4K20me3 levels were significantly correlated in a univariate and multivariate model with bladder cancer-specific mortality after radical cystectomy in patients with MIBC. CONCLUSION • Global histone methylation levels may help to identify patients with bladder cancerwith poor prognosis after radical cystectomy. [ABSTRACT FROM AUTHOR]

Abstract: Gene silencing by epigenetic mechanisms is frequent in prostate cancer (PCA). The link between DNA hypermethylation and histone modifications is not completely understood. We chose the GSTP1 gene which is silenced by hypermethylation to analyze the effect of the histone deacetylase inhibitor depsipeptide on DNA methylation and histone modifications at the GSTP1 promoter site. Prostate cell lines (PC-3, LNCaP, and BPH-1) were treated with depsipeptide; apoptosis (FACS analysis), GSTP1 mRNA levels (quantitative real-time PCR), DNA hypermethylation (methylation-specific PCR), and histone modifications (chromatin immunoprecipitation) were studied. Depsipeptide induced apoptosis in PCA cells, but not a cell cycle arrest. Depispeptide reversed DNA hypermethylation and repressive histone modifications (reduction of H3K9me2/3 and H3K27me2/3; increase of H3K18Ac), thereby inducing GSTP1 mRNA re-expression. Successful therapy requires both, DNA demethylation and activating histone modifications, to induce complete gene expression of epigenetically silenced genes and depsipeptide fulfils both criteria. [Copyright &y& Elsevier]

Background: MicroRNA expression is altered in cancer cells, and microRNAs could serve as diagnostic/prognostic biomarker for cancer patients. Our study was designed to analyze circulating serum microRNAs in patients with renal cell carcinoma (RCC). Methodology/Principal Findings: We first explored microRNA expression profiles in tissue and serum using TaqMan Low Density Arrays in each six malignant and benign samples: Although 109 microRNAs were circulating at higher levels in cancer patients' serum, we identified only 36 microRNAs with up-regulation in RCC tissue and serum of RCC patients. Seven candidate microRNAs were selected for verification based on the finding of up-regulation in serum and tissue of RCC patients: miR-7-1*, miR-93, miR-106b*, miR-210, miR-320b, miR-1233 and miR-1290 levels in serum of healthy controls (n = 30) and RCC (n = 33) patients were determined using quantitative real-time PCR (TaqMan MicroRNA Assays). miR-1233 was increased in RCC patients, and thus validated in a multicentre cohort of 84 RCC patients and 93 healthy controls using quantitative real-time PCR (sensitivity 77.4%, specificity 37.6%, AUC 0.588). We also studied 13 samples of patients with angiomyolipoma or oncocytoma, whose serum miR-1233 levels were similar to RCC patients. Circulating microRNAs were not correlated with clinical-pathological parameters. Conclusions/Significance: MicroRNA levels are distinctly increased in cancer patients, although only a small subset of circulating microRNAs has a tumor-specific origin. We identify circulating miR-1233 as a potential biomarker for RCC patients. Larger-scaled studies are warranted to fully explore the role of circulating microRNAs in RCC. [ABSTRACT FROM AUTHOR]

Abstract: The presence of small amounts of circulating DNA in plasma was demonstrated 60 years ago. Since then, cell-free DNA has been tested for quantity, fragmentation pattern, and tumor-specific sequences in patients with various malignancies. Recent studies have shown that cell-free DNA levels are distinctly increased in most patients with prostate cancer (PCA) and that the DNA fragmentation pattern is different from healthy individuals and patients with benign prostate disease. The origin of this circulating DNA remains largely unknown, but it is established that a small fraction of the DNA is derived from the tumor itself, and genetic (allelic imbalances) and epigenetic (DNA methylation) alterations are regularly detected in patients with PCA. The detection of increased DNA levels and tumor-specific DNA sequences may provide diagnostic and prognostic information. The recent findings in the emerging field of cell-free DNA will be discussed. [Copyright &y& Elsevier]

Scars arise in the late phase of wound healing and are characterized by fibroplasia. Previous controversial studies have discussed the regenerative wound healing capacity of Murphy Roths Large (MRL) mice. The aim of this study was to investigate the mechanisms of improved wound healing in a skin transplantation model. Skin grafts from MRL and haplotypically identical B10.BR mice were cross-transplanted. At day 10, B10.BR and MRL grafts on B10.BR recipients deposited collagen and showed severe apoptosis. Grafts of MRL recipients were not affected by such alterations and showed an enhanced healing progress. They were characterized by higher partial pressure of tissue oxygen, increased microcirculation, exceptionally intense neovascularization, and a blunted inflammatory response. This phenotype was accompanied by increased vascular endothelial growth factor expression, augmented by enhanced signal transducer and activator of transcription 3 (STAT3) phosphorylation. These effects were combined with a decreased STAT1 expression and phosphorylation. STAT1 pattern variation was associated with decreased Smad7 levels. Furthermore, MRL recipients showed improved stem cell recruitment to the wound area. The basic accelerated wound healing mechanism in MRL mice found in this skin transplantation model is improved engraftment; this is based on enhanced neovascularization and reduced inflammation. These effects are most likely due to higher vascular endothelial growth factor levels and changes in the STAT/Smad signal pathway, which may enhance transforming growth factor-β signaling, reducing proinflammatory responses. [ABSTRACT FROM AUTHOR]

OBJECTIVE To analyse the diagnostic and prognostic value of cell-free mitochondrial (mt)DNA in patients with testicular cancer, as increased levels of cell-free circulating mtDNA have been reported in patients with cancer. PATIENTS, SUBJECTS AND METHODS In all, 74 patients with testicular cancer (seminoma in 39, nonseminoma in 35) and 35 healthy individuals were included in the study. Circulating DNA was isolated from 1 mL of serum. A quantitative real-time polymerase chain reaction was used to analyse the levels of a 79-bp (mtDNA-79) and 220 bp (mtDNA-220) fragment of the mitochondrial specific 16S-RNA. The mtDNA integrity was expressed as the ratio of mtDNA-220 to mtDNA-79. RESULTS mtDNA-79 and mtDNA-220 levels were significantly ( P < 0.001) greater in patients with testicular cancer than in healthy individuals. The mtDNA integrity was similar in patients and healthy controls ( P = 0.435). Receiver operator characteristic curve analysis showed that cell-free mtDNA (mtDNA-79) levels distinguished, with a sensitivity of 59.5% and a specificity of 94.3%, between patients and healthy individuals (area under curve, 0.787). Also, mtDNA-79 levels could be used to distinguish between patients (31) with conventional markers (α-fetoprotein, human chorionic gonadotrophin, placental alkaline phosphatase and lactate dehydrogenase) within normal ranges and healthy individuals, with a sensitivity of 64.5% and specificity of 91.4% (area under curve 0.797). Cell-free mtDNA levels were not correlated with any clinicopathological variable (pT stage, lymph node invasion, vascular invasion, clinical stage, International Germ Cell Cancer Collaborative Group classification, tumour markers; all P > 0.05). CONCLUSION Cell-free mtDNA levels are greater in patients with testicular cancer and might provide valuable information for managing patients with testicular anomalies, especially those with normal levels of established tumour markers. [ABSTRACT FROM AUTHOR]

OBJECTIVE To investigate the role of circulating mitochondrial DNA (mtDNA) in patients with localized prostate cancer, as recent reports show that patients with advanced cancer have increased levels of mtDNA. PATIENTS AND METHODS DNA was isolated from the serum of 100 patients with prostate cancer and 18 with benign prostate hyperplasia (BPH). A quantitative real-time polymerase chain reaction was used to amplify 79 bp and 230 bp fragments of the mitochondrial 16s-RNA gene, the short fragment representing total mtDNA, including mtDNA truncated by apoptosis, and the long fragment representing mostly mtDNA from other cell death entities. mtDNA integrity was defined as the ratio of long to short mtDNA fragments. RESULTS The short and long mtDNA levels, and mtDNA integrity, were similar in patients with BPH and cancer ( P = 0.940, 0.211 and 0.441, respectively), and were not correlated with clinical or pathological variables, e.g. age, prostate-specific antigen (PSA) level, cT stage, pT stage, seminal vesicle infiltration, lymph node invasion, or Gleason score ( P = 0.075 to 0.961). However, patients with high levels of short mtDNA (>75th percentile) had a greater risk of PSA progression and this variable was the strongest predictor of PSA recurrence in a multivariate Cox analysis ( P = 0.023; hazard ratio 0.31; 95% confidence interval 0.113–0.851). CONCLUSION Circulating mtDNA levels did not distinguish between patients with prostate cancer or BPH. However, there was a significant increase in short mtDNA fragments in patients with early PSA recurrence after radical prostatectomy. [ABSTRACT FROM AUTHOR]

Abstract: Patients with malignancies often possess increased concentrations of cell-free serum DNA. In this study, we investigated serum DNA levels in each 45 patients with bladder cancer (BCA) undergoing radical cystectomy and with benign prostate hyperplasia. A quantitative real-time PCR was used to amplify a 124bp (PTGS2; mostly apoptotic origin) and a 271bp (Reprimo; mostly necrotic origin) DNA fragment. Changes in the origin of DNA fragments were specified as the Apoptosis Index (AI, ratio of 124bp/271bp fragments). Small and large fragments were increased (p <0.001 and p =0.041) in BCA patients. The AI increase suggests that DNA fragmentation was mostly (p <0.001) caused by apoptosis. High levels of small DNA fragments distinguished between BCA and BPH with high sensitivity (96%) and moderate specificity (62%). DNA levels and the AI were not correlated with clinicopathological parameters. However, an increased AI was correlated with BCA-specific mortality in a multivariate analysis (p =0.011) indicating that the AI is an independent prognostic factor. Thus, cell-free DNA seems to be a useful prognostic marker in patients with BCA. [Copyright &y& Elsevier]

Objectives: CpG island hypermethylation causes gene silencing and could be decisive in prostate carcinogenesis and progression. We investigated its role at multiple gene sites during prostate carcinogenesis. Methods: A quantitative, methylation-specific polymerase chain reaction was used to analyze the hypermethylation patterns at nine gene loci (Annexin2, APC, EDNRB, GSTP1, PTGS2, MDR1, RARbeta, Reprimo, and TIG1) in 80 patients with prostate cancer (PCa) and 26 patients with benign prostatic hyperplasia (BPH). Results: Hypermethylation was more frequent in PCa than in BPH tissues (EDNRB, 100% versus 88%; TIG1, 96% versus 12%; RARbeta, 95% versus 35%; GSTP1, 93% versus 15%; APC, 80% versus 50%; MDR1, 80% versus 31%; PTGS2, 68% versus 15%; Reprimo, 59% versus 19%; and Annexin2, 4% versus 0%). TIG1 and GSTP1 hypermethylation distinguished between PCa and BPH with a specificity of greater than 85% and sensitivity of greater than 93%. Hypermethylation at a single gene locus did not correlate with any clinicopathologic variables. In contrast, hypermethylation at two genes (eg, APC and TIG1, APC and GSTP1, APC and PTGS2, APC or MDR, GSTP1 or PTGS2) correlated significantly with the pathologic stage and/or Gleason score (P = 0.033 to 0.045). Hypermethylation at APC and Reprimo, as well as DNA hypermethylation at more than five genes, correlated significantly with the rate of prostate-specific antigen recurrence after radical prostatectomy (P = 0.0078 and P = 0.0074, respectively). Conclusions: Our results have confirmed that the hypermethylation patterns are helpful in the diagnosis and prognosis of PCa. Increases in CpG island hypermethylation at multiple gene sites occur during PCa progression and indicate early biochemical recurrence after radical prostatectomy. [Copyright &y& Elsevier]

Background: Little is known about the local accumulation and function of immune cells in peritoneal fluid after elective surgery of the upper and lower gastrointestinal tract. Our study was designed to investigate whether systemic immune cell response mirrors the local response. We focused on the cytokines interleukin (IL)-6 and tumor necrosis factor (TNF)-α and on monocytes, natural killer (NK) cells, and T cells that play an important role in eliciting the innate and adaptive immune response. Methods: Blood samples were taken prospectively from 25 patients 24 h before surgery, as well as 24 h and 48 h afterward. Abdominal drainage fluids were collected intraoperatively 1 h after the abdomen was opened and 24 h and 48 h postoperatively. Apart from the white blood cells, intracellular T-helper-cell (TH1/TH2) cytokine production (interferon-γ, IL-2, IL-4, IL-13) and HLA-DR on monocytes were measured by four-color flow cytometry, IL-6, and TNF-α with the fast immunoluminescence method. Results: Cells of the innate immune system (NK cells, monocytes, NK-T cells, CD5+ B cells) rapidly decreased in abdominal fluids (P &#60; 0.05: +24 h; +48 h) after surgery, which was paralleled by a concomitant decline in peripheral blood. The percentage of abdominal interferon-γ, IL-2, IL-4, and IL-13-producing TH cells increased in a way that distinctly counteracted the decrease of the natural immune cells. HLA-DR expression on monocytes in peripheral blood declined significantly (P &#60; 0.05: +24 h; +48 h). In contrast, monocytes in abdominal fluids had high HLA-DR expression. Furthermore, abdominal fluids contained significantly higher concentrations of TNF-α (P &#60; 0.05: +24 h; +48 h) and IL-6 (P &#60; 0.05: +24 h) compared with peripheral blood. Conclusions: Specific immune cell recruitment and cytokine production play an important role in post-trauma events. Measuring distinct local immune cell repertoires and cytokines provides answers as to how the different phases of postoperative immune events proceed. The evaluation of the local response may provide additional criteria for the evaluation of operative trauma. This knowledge may be helpful in detecting postoperative pathological aberrancies. [Copyright &y& Elsevier]

Around 1-2% of the net C production by land vegetation is re-emitted to the atmosphere as Biogenic Volatile Organic Compounds (BVOCs). Besides de small contribution to C balance, BVOCs play important roles in biogeochemical and physical processes, as well as in biosphere-atmosphere interactions, by influencing atmospheric oxidative capacity and potentially forming secondary organic aerosols. Isoprene and monoterpenes account for the largest amount of global fluxes, but emission patterns by species are still puzzling and hinder accurate modeling. Estimates are that only about 30% of tree species can emit isoprene, and that this capacity has evolved and been lost multiple times during evolution. Isoprene emission mitigates heat and drought stress in leaves, and hypotheses suggest that emission capacity would be associated with sun-exposed, high canopy trees. All tree species are capable of emitting monoterpenes, but there are major differences in compound structural diversity and magnitude of emissions. Monoterpenes are usually associated with defense strategies against herbivory, and higher emission rates, storage and compound diversity would potentially be observed in perennial, shade-tolerant trees. In terms of plant C production costs, monoterpene emissions and storage should be more costly compared to isoprene emissions. We hypothesize that there is a trade-off relationship between isoprene and monoterpene emission capacities, and that this trade-off is linked to the fast-slow plant economics spectrum, which groups species according to the allocation of carbon towards growth (fast trees) or survival (slow trees) strategies. We predict that the capacity to emit isoprene tends to appear in fast trees, whereas higher total C costs, emission rates, storage and diversity of monoterpenes are more associated with slow trees. We measured isoprenoid leaf flux, storable isoprenoid content, photosynthetic rate, stomatal conductance and plant functional traits of i) leaf (leaf area, specific leaf area, leaf dry matter content, leaf thickness, force to tear, stomatal size and density, leaf vessel density), ii) wood (stem and twig wood density, stem and twig wood water content, active xylem depth), and iii) whole-plant (proportions of basal area to total height, active xylem area to basal area, crown area/volume to basal area) for 30 Amazonian tree species (3 individuals per species) of 17 families. Measurements were carried out from October to December 2018 at the Amazonian Tall Tower Observatory (ATTO) experimental plots in the central Brazilian Amazon. We will present multivariate relationships between isoprenoids and plant functional traits that reveal the influence of resource allocation strategies on the mysterious evolutionary pattern of emissions. A better mechanistic understanding will improve emission estimates for unmeasured species, and thereby improve the scaling of emission estimates across tropical forests. To the authors' knowledge, this is the first research that approaches isoprenoid emission patterns from the perspective of a broad set of plant functional traits. Considering that global climate change will possibly affect species turnover and composition of Amazonian tree communities and that BVOC emissions are dependent on species diversity and strategies, this new approach may shed light on future patterns of global BVOC fluxes and hence impacts in ecosystem-atmosphere dynamics. [ABSTRACT FROM AUTHOR]

Objectives: To analyze circulating microRNAs (miRNA) in serum as non-invasive biomarker in patients with localized prostate cancer (PCA), benign prostate hyperplasia (BPH) and healthy individuals (HI). Methods: Total RNA was isolated from serum samples and the circulating levels of different RNA species (miRNA, miR-16; small nuclear RNA, RNU1A-1; messenger RNA, HPRT1), as well as of 4 oncogenic miRNAs (miR-26a, miR-32, miR-195, miR-let7i), were determined using a quantitative real-time polymerase chain reaction. We also evaluated miRNA levels in a second cohort of 10 PCA patients in cancer/nonmalignant tissue, and pre- and post-prostatectomy serum samples. Results: The levels of miR-16 and RNU1A-1were reliably measured, whereas HPRT1 levels were often below the detection limit of our assay. Circulating oncogenic miRNA levels were different, and especially the miR-26a level allowed sensitive (89%) discrimination of PCA and BPH patients at a moderate specificity (56%; area under the curve [AUC]: 0.703); the analysis of oncogenic miRNAs in combination increased the diagnostic accuracy (sensitivity: 78.4%; specificity: 66.7%; AUC: 0.758). Despite the low number of patients limiting the statistical power of the study, we observed correlations with clinical-pathologic parameters: miR-16, miR-195, and miR-26a were significantly correlated with surgical margin positivity; miR-195 and miR-let7i were significantly correlated with the Gleason score. Tissue miRNA levels were correlated with preprostatectomy miRNA levels in serum, and serum miRNA decreased after prostatectomy, thereby indicating tumor-associated release of miRNA. Conclusions: Tumor-associated miRNAs in serum allow noninvasive discrimination of PCA and BPH. [ABSTRACT FROM AUTHOR]

Gene silencing by promoter DNA hypermethylation is common in PCA. The role of HXKy methylation as an up-/downstream or parallel silencing mechanism is not yet clear. We analyzed HXKy methylation at gene sites that are aberrantly DNA-methylated in PCA (i.e. GSTP1, RARβ, APC and PTGS2). Cancerous (PC3, DU145, LNCaP) and benign prostate hyperplasia (BPH1) cell lines were studied using chromatin immunoprecipitation and methylation specific PCR. Gene expression was analyzed using quantitative RT-PCR. In PCA, histone modifications characteristic of gene repression (di-/tri-methylated H3K9, H3K27, H4K20) were predominantly observed at the studied gene sites and paralleled aberrant DNA hypermethylation. In BPH, no aberrant DNA hypermethylation and mostly histone modifications supporting active gene expression (H3K4, H3K79 di-/tri methylation) were seen at the respective gene sites. Mono-methylated H3K4 and H3K9 modifications that have moderate effects on gene expression were frequent in both PCA and BPH. PC3 cells grown in the presence of the DNA methyltransferase inhibitor 5-aza-2′-deoxycytidine (aza) resulted in DNA demethylation and moderate gene reexpression whereas repressive HXKy di-methylation increased 1.5-3 fold and tri-methylation was mostly unchanged. Removal of aza reverses gene expression. These findings suggest that HXKy methylation is upstream to DNA hypermethylation in PCA. [ABSTRACT FROM AUTHOR]

Cancer patients have increased levels of serum DNA fragments (SDF) compared to most patients with non-malignant disease and healthy individuals. In PCA, DNA hypermethylation is also frequently observed (>80%) at promoter CpG islands of GSTP1, PTGS2 and TIG1. Therefore we analysed hypermethylation of these genes in SDF from patients with PCA (n=168) and benign prostate hyperplasia (BPH; n=42) and controls (n=11) for information about the cellular origin of DNA. SDF concentrations and the hypermethylated fractions were analysed using a methylation-sensitive restriction enzyme based real-time PCR. SDF levels were significantly increased in PCA vs BPH vs controls (65 vs 20 vs 7 ng/mL; p<0.001). Hypermethylation of GSTP1, TIG1 or PTGS2 was more frequently detected in PCA than in BPH patients and controls (overall 47% vs 8% vs 0%; p<0.001), but did not exceed 10% of total DNA. SDF levels distinguished PCA from BPH patients with a sensitivity of 83% and specificity of 69%. The diagnostic significance of SDF was greater than that of hypermethylation data. In conjunction with SDF, hypermethylation numbers did not contribute any further information. Both SDF levels and the hypermethylation status were not correlated to advanced tumor stage or grade (p>0.05). In summary SDF is a promising diagnostic marker in PCA. It does not primarily originate from tumor cells but from apoptotic and necrotic cells surrounding the tumor. [ABSTRACT FROM AUTHOR]