Prognostic significance of primary-tumor extension, stage and grade of nuclear differentiation in patients with renal cell carcinoma

Surgery remains the preferred therapy for renal cell carcinoma. The various adjunctive or complementary therapies currently yield disappointing results. Identifying reliable prognostic factors could help in selecting patients most likely to benefit from postoperative adjuvant therapies. We reviewed the surgical records of 78 patients who had undergone radical nephrectomy with lymphadenectomy for renal cell carcinoma, matched for type of operation and histology. According to staging (TNM), 5.1% of the patients were classified as stage I, 51.3% as stage II, 29.5% as stage III and 14.5% as stage IV. Of the 78 patients 40 were T2N0 and 21 T3aN0. Tumor grading showed that 39.7% of the patients had well-differentiated tumors(G1), 41.1% moderately-differentiated (G2), and 19.2% poorly-differentiated tumors (G3). Overall actuarial survival at 5 and 10 years was 100% for stage 1; 91.3% at 5 years and 83.1% at 10 years for stage II; 45.5% and 34.1% for stage III; and 29.1% and nil for stage IV (stage II vs stage III p = 0.0001). Patients with tumors confined to the kidney (pT2N0) had better 5- and 10-year survival rates than patients with tumors infiltrating the perirenal fat (pT3aN0) (p = 0.000006). Survival differed according to nuclear grading (G1 vs G3 ; p = 0.000005; G2 vs G3; p = 0.0009). In conclusion our review identified tumor stage, primary-tumor extension, and the grade of nuclear differentiation as reliable prognostic factors in patients with renal cell carcinomas

Paralog-Specific Patterns of Structural Disorder and Phosphorylation in the Vertebrate SH3–SH2–Tyrosine Kinase Protein Family

One of the largest multigene families in Metazoa are the tyrosine kinases (TKs). These are important multifunctional proteins that have evolved as dynamic switches that perform tyrosine phosphorylation and other noncatalytic activities regulated by various allosteric mechanisms. TKs interact with each other and with other molecules, ultimately activating and inhibiting different signaling pathways. TKs are implicated in cancer and almost 30 FDA-approved TK inhibitors are available. However, specific binding is a challenge when targeting an active site that has been conserved in multiple protein paralogs for millions of years. A cassette domain (CD) containing SH3–SH2–Tyrosine Kinase domains reoccurs in vertebrate nonreceptor TKs. Although part of the CD function is shared between TKs, it also presents TK specific features. Here, the evolutionary dynamics of sequence, structure, and phosphorylation across the CD in 17 TK paralogs have been investigated in a large-scale study. We establish that TKs often have ortholog-specific structural disorder and phosphorylation patterns, while secondary structure elements, as expected, are highly conserved. Further, domain-specific differences are at play. Notably, we found the catalytic domain to fluctuate more in certain secondary structure elements than the regulatory domains. By elucidating how different properties evolve after gene duplications and which properties are specifically conserved within orthologs, the mechanistic understanding of protein evolution is enriched and regions supposedly critical for functional divergence across paralogs are highlighted

Predictive value of hematological and phenotypical parameters on postchemotherapy leukocyte recovery

Background: Grade IV chemotherapy toxicity is defined as absolute neutrophil count <500/μL. The nadir is considered as the lowest neutrophil number following chemotherapy, and generally is not expected before the 7th day from the start of chemotherapy. The usual prophylactic dose of rHu-G-CSF (Filgrastim) is 300 μg/day, starting 24-48 h after chemotherapy until hematological recovery. However, individual patient response is largely variable, so that rHu-G-CSF doses can be different. The aim of this study was to verify if peripheral blood automated flow cytochemistry and flow cytometry analysis may be helpful in predicting the individual response and saving rHu-G-CSF. Methods: During Grade IV neutropenia, blood counts from 30 cancer patients were analyzed daily by ADVIA 120 automated flow cytochemistry analyzer and by Facscalibur flow cytometer till the nadir. "Large unstained cells" (LUCs), myeloperoxidase index (MPXI), blasts, and various cell subpopulations in the peripheral blood were studied. At nadir rHu-G-CSF was started and 81 chemotherapy cycles were analyzed. Cycles were stratified according to their number and to two dose-levels of rHuG-CSF needed to recovery (300-600 vs. 900-1200 μg) and analyzed in relation to mean values of MPXI and mean absolute number of LUCs in the nadir phase. The linear regressions of LUCs % over time in relation to two dose-levels of rHu-G-CSF and uni-multivariate analysis of lymphocyte subpopulations, CD34+ cells, MPXI, and blasts were also performed. Results: In the nadir phase, the increase of MPXI above the upper limit of normality (>10; median 27.7), characterized a slow hematological recovery. MPXI levels were directly related to the cycle number and inversely related to the absolute number of LUCs and CD34 +/CD45+ cells. A faster hematological recovery was associated with a higher LUC increase per day (0.56% vs. 0.25%), higher blast (median 36.7/μL vs. 19.5/μL) and CD34+/CD45+ cell (median 2.2/μL vs. 0.82/μL) counts. Conclusions: Our study showed that some biological indicators such as MPXI, LUCs, blasts, and CD34 +/CD45+ cells may be of clinical relevance in predicting individual hematological response to rHu-G-CSF. Special attention should be paid when nadir MPXI exceeds the upper limit of normality because the hematological recovery may be delayed. © 2009 Clinical Cytometry Society

Acrylamide levels in cooked rice, tomato sauces and some fast food on the Italian market

This study reports the results of evaluation of acrylamide levels in some foods that are common on the Italian market. Three foods commonly found in the national diet (rice, tomato sauce and fast food), were examined with the gas chromatograph (GC)/mass spectrometer (MS) analytical method. Results show that rice differs from risotto with respect to acrylamide levels: values of less than 50 \u3bcg/kg, for boiled rice, increase to 113 \u3bcg/kg when various ingredients are added to produce risotto. Similar results were found for tomato sauce on the Italian market: acrylamide values were less than 50 \u3bcg/kg for simple tomato sauce, to 124 \u3bcg/kg when other ingredients such as olives and capers were added. Fast foods (e.g., fried potatoes) contained the highest observed acrylamide levels, probably from cooking methods and acrylamide-rich precursors. For two fried potatoes of the same type, very differentiated values resulted (136 and 294 \u3bcg/kg)

Characterization of the biogenic volatile organic compounds (BVOCs) and analysis of the PR1 molecular marker in Vitis vinifera L. inoculated with the nematode Xiphinema index

Upon pathogen attack, plants very quickly undergo rather complex physico-chemical changes, such as the production of new chemicals or alterations in membrane and cell wall properties, to reduce disease damages. An underestimated threat is represented by root parasitic nematodes. In Vitis vinifera L., the nematode Xiphinema index is the unique vector of Grapevine fanleaf virus, responsible for fanleaf degeneration, one of the most widespread and economically damaging diseases worldwide. The aim of this study was to investigate changes in the emission of biogenic volatile organic compounds (BVOCs) in grapevines attacked by X. index. BVOCs play a role in plant defensive mechanisms and are synthetized in response to biotic damages. In our study, the BVOC profile was altered by the nematode feeding process. We found a decrease in \u3b2-ocimene and limonene monoterpene emissions, as well as an increase in \u3b1-farnesene and \u3b1-bergamotene sesquiterpene emissions in nematode-treated plants. Moreover, we evaluated the PR1 gene expression. The transcript level of PR1 gene was higher in the nematode-wounded roots, while in the leaf tissues it showed a lower expression compared to control grapevines

GLUT1 and LOX inhibitors as perspective anticancer agents tackling glucose avidity and ECM remodeling in tumors

Introduction Most cancers have large hypoxic regions, which display an increase of the glycolytic metabolism leading to the production of lactate, providing cancer cells with adequate amounts of energy and anabolites. To this end, tumor cells generally overexpress glucose transporters (GLUTs), in particular GLUT1, which results in an increased uptake of glucose to support their less efficient energy production (Warburg effect). Therefore, therapeutic interventions aimed at reducing cancer glycolysis may be implemented by several strategies, including the development of inhibitors of glucose transporters. Furthermore, extracellular matrix (ECM) remodeling is one of the key processes preluding metastatic invasion, and it is promoted by several effectors, such as lysyl oxidase (LOX), an enzyme commonly involved in extracellular matrix maturation. LOX is up-regulated by HIF-1 and plays a critical role in the development of metastasis. Therefore, LOX inhibitors may represent an additional and innovative strategy for the treatment and the prevention of metastatic cancer. Methods We have developed various classes of compounds that are able to interfere with GLUTs (Granchi et al. 2015, Tuccinardi et al. 2013) and LOX (Granchi et al. 2009) by molecular design and chemical synthesis. Their effect on cell proliferation, apoptosis, migration and other key determinants of activity were evaluated by sulforhodamine-B and luciferase assays, FACS, wound-healing assay, and Quantitative PCR. The studies were performed in seven PDAC cells, including five primary-cell-cultures and 3D co-cultures with human stellate cells, in normoxic and hypoxic conditions. Results The IC50s of the tested compounds ranged from 13.9 to 32.0 μM after 72-hour exposure. Notably, these compounds were still active in 3D co-cultures of these tumor cells with pancreatic stellate cells, which showed increased resistance to gemcitabine and are more representative of the dense stromal compartment with core hypoxic areas of this tumor type, as detected by immunohistochemical stainings. Remarkably, one compound (PGL-14) showed a synergistic interaction with gemcitabine, increasing apoptosis induction and accumulation of ROS. Furthermore, the combination of these drugs reduced cell migration and enhanced in vitro sensitivity to anoikis, suggesting the ability of these compounds to inhibit metastasis. Discussion GLUT1 inhibitors were more active in hypoxia, but still active also in normoxia. Conversely, we did not detect cytotoxic effects using the LOX-inhibitors in normoxia (at concentration until 50 μM) since they were designed as bioreductively activated prodrugs, which are therefore activated only under hypoxic conditions. However, at O2 tension of 1%, IC50s were below 10 μM. As reported previously, LOX inhibition was associated with reduction of the mRNA levels of fibronectin, suggesting that it might also have impact on the interaction of tumor cells with the stroma that are mediated by integrins and fibronectin, regulating tissue stiffness (Coppola et al. 2017). Conclusion Interventions aimed at blocking the glycolytic activity or the extracellular matrix remodeling of tumors by means of newly designed molecules proved to exert a synergistic effect with clinically approved drugs, such as gemcitabine. These results seem to support the strategy of the simultaneous GLUT/LOX-inhibition in order to further sensitize hypoxic cancer portions to chemotherapy. Bibliography C. Granchi, et al. ChemMedChem 2009, 4, 1590-1594. C. Granchi, et al. ChemMedChem 2015, 10, 1892-1900. T. Tuccinardi, et al. Bioorg. Med. Chem. Lett. 2013, 23, 6923-6927. Coppola S, et al. Drug Resist Updat. 2017, 31, 43-51

Sirtuin 1-Activating Compounds: Discovery of a Class of Thiazole-Based Derivatives

Sirtuin 1 (SIRT1) is a NAD+-dependent deacetylase implicated in various biological and pathological processes, including cancer, diabetes, and cardiovascular diseases. In recent years, SIRT1-activating compounds have been demonstrated to exert cardioprotective effects. Therefore, this enzyme has become a feasible target to treat cardiovascular diseases, and many SIRT1 activators, of a natural or synthetic origin, have been identified. In the present work, we developed thiazole-based SIRT1 activators, which showed remarkably higher SIRT1 activation potencies compared with those of the reference compound resveratrol when tested in enzymatic assays. Thiazole 8, a representative compound of this series, was also subjected to further pharmacological investigations, where it was proven to reduce myocardial damage induced by an in vivo occlusion/reperfusion event, thus confirming its cardioprotective properties. In addition, the cardioprotective effect of compound 8 was significantly higher than that of resveratrol. Molecular modeling studies suggest the binding mode of these derivatives within SIRT1 in the presence of the p53-AMC peptide. These promising results could pave the way to further expand and optimize this chemical class of new and potent SIRT1 activators as potential cardioprotective agents

First data on the antimicrobial activity of Yucca filamentosa L. bark exytacts

The aim of this study was to investigate the antimicrobial activity of two Yucca filamentosa L. extracts produced by hydro-alcoholic extraction under different operational conditions. The extracts were concentrated and de-alcoholised, and were added to media to test their effects on target microorganisms. Both extracts displayed equivalent specific growth inhibitory activity against Saccharomyces cerevisiae. No other antimicrobial activity was found. This anti-fermentative activity may open the doors to new possibilities for using Yucca filamentosa L. extracts in the food industry. These findings are of particular interest, since yucca extracts are already classified as being generally recognized as safe (GRAS)

Dynamic of microbial community during distinct stages of organic matter decomposition in sediments from mangroves of São Paulo state.

Abstract: Mangroves are ecosystems located at coastline areas in tropical and subtropical conditions that are under tide influences. They are recognized as productive regions and have a high export of organic matter (OM) for the adjacent coastal waters. Bacterial communities present in these environments perform primordial functions on the cycling of nutrients and thereby on maintaining your important processes. Therefore, the purpose of this study was to assess the dynamics and phylogenetic diversity of the bacterial community in threes of São Paulo mangroves, during different stages of decomposition plant material. At the same time, the intention was to correlate this event with the emission of greenhouse gases (GHG). Microcosms were constructed with leaf discs of Avicennia schaueriana (A), Laguncularia racemosa (L), Rhizophora mangle (R) were arranged on sediment and water, which were collected from the following Mangroves: Bertioga with contamination (BC), Bertioga (B) and Cananeia (C). These microenvironments were stored at 24 °C and monitored for 60 days. The measurements of gases and DNA extraction were performed at the 7, 15, 30 and 60 days. The collected air samples were analyzed in triplicate in chromatograph model GC-2014 greenhouse gas. The obtained DNA was subjected to PCR reactions aiming the sequencing in large scale of the gene that encoding the ribosomal rRNA Domain Bacteria by Ion Torrent. The assessment of communities dynamics was made with Qiime. Regarding CO2 emissions, the values observed for AB7, RB7 and RBC7 were higher than the other treatments. For N2O the samples AB, ABC7 and RBC7 showed the higher rates. The CH4 gas obtained a homogeneous result for all samples, seeing no significant difference. The bacterial community dynamics showed that treatments: Bertioga, Avicennia schaueriana and decomposition time 60 days, presented a higher clustering within their samples, according to the standard of similarity observed in them

Change in enzymatic activity of microbial communities on the seasonal differences in soils the of biomes caatinga and atlantic forest.

Abstract: The average global temperature has increased and will continue increasing, with forecast of up to 5.8° C by the end of this century as a result of the increase in the concentration of greenhouse gases in the atmosphere. Due to these increased temperature, some biomes may introduce features of arid and semi-arid environments in the coming years. The patterns of water availability in the soil can select groups of microorganisms with physiological strategies to tolerate these seasonal changes. Nutrient availability for plants becomes limited when the aqueous part of the soil is low in the dry period. On the other hand, in irrigated soils the concentration of nutrients available associated with mineralization increases rapidly. Environmental indicators such as seasonal variations in the enzymatic activity, the abundance of microorganisms and soil physical and chemical characteristics, are useful to measure and analyze the change of the functionality soil microbiota. This work was aimed to study the seasonal changes of enzymatic activity (dehydrogenase, acid phosphatase and alkaline, arylsulfatase, phytase, cellulase, amylase and urease) of microbial communities in soils of Caatinga and Atlantic Forest. The soils were collected in Bertioga-SP, Cananéia-SP, Juazeiro-BA and Petrolina-PE. The samples of soil were sifted and separated into 18 glass bottles of 200 mL containing 50 g of each soil. The bottles were incubated at 28° C varying moisture in 60% and 30% every 14 days corresponding to three seasonal cycles. The sequencing of 16S ribosomal RNA gene (rRNA) of samples was performed using the Ion Sequencing Kit 400 (Life Technologies). The dynamics of the bacterial communities had influences of seasonal changes, especially when compared the biogeographic patterns. The effect of moisture was significantly different (p < 0.05) only to the activity of alkaline phosphatase and phytase enzymes. The activity of all enzymes, except phytase, was important to discriminate the soils between Caatinga and Atlantic Forest biomes. Seasonality had an effect on the enzymatic activity that was related with the functional change in the soil microbiota