Molecular Determinants of Soft Tissue Sarcoma Immunity: Targets for Immune Intervention

Soft tissue sarcomas (STSs) are a family of rare malignant tumors encompassing more than 80 histologies. Current therapies for metastatic STS, a condition that affects roughly half of patients, have limited efficacy, making innovative therapeutic strategies urgently needed. From a molecular point of view, STSs can be classified as translocation-related and those with a heavily rearranged genotype. Although only the latter display an increased mutational burden, molecular profiles suggestive of an “immune hot” tumor microenvironment are observed across STS histologies, and response to immunotherapy has been reported in both translocation-related and genetic complex STSs. These data reinforce the notion that immunity in STSs is multifaceted and influenced by both genetic and epigenetic determinants. Cumulative evidence indicates that a fine characterization of STSs at different levels is required to identify biomarkers predictive of immunotherapy response and to discover targetable pathways to switch on the immune sensitivity of “immune cold” tumors. In this review, we will summarize recent findings on the interplay between genetic landscape, molecular profiling and immunity in STSs. Immunological and molecular features will be discussed for their prognostic value in selected STS histologies. Finally, the local and systemic immunomodulatory effects of the targeted drugs imatinib and sunitinib will be discussed.     

Desorption electrospray ionization then MALDI mass spectrometry imaging of lipid and protein distributions in single tissue sections

Imaging mass spectrometry (MS) is a powerful technique for mapping the spatial distributions of a wide range of chemical compounds simultaneously from a tissue section. Co-localization of the distribution of individual molecular species, including particular lipids and proteins, and correlation with the morphological features of a single tissue section are highly desirable for comprehensive tissue analysis and disease diagnosis. We now report on the use, in turn, of desorption electrospray ionization (DESI), matrix assisted laser desorption ionization (MALDI), and then optical microscopy to image lipid and protein distributions in a single tissue section. This is possible through the use of histologically compatible DESI solvent systems, which allow for sequential analyses of the same section by DESI then MALDI. Hematoxylin and eosin (H&E) staining was performed on the same section after removal of the MALDI matrix. This workflow allowed chemical information to be unambiguously matched to histological features in mouse brain tissue sections. The lipid sulfatide (24:1), detected at m/z 888.8 by DESI imaging, was colocalized with the protein MBP isoform 8, detected at m/z 14117 by MALDI imaging, in regions corresponding to the corpus callosum substructure of the mouse brain, as confirmed in the H&E images. Correlation of lipid and protein distributions with histopathological features was also achieved for human brain cancer samples. Higher tumor cell density was observed in regions demonstrating higher relative abundances of oleic acid, detected by DESI imaging at m/z 281.4, and the protein calcyclin, detected by MALDI at m/z 10085, for a human glioma sample. Since correlation between molecular signatures and disease state can be achieved, we expect that this methodology will significantly enhance the value of MS imaging in molecular pathology for diagnosis.     

Extension of Tosèla cheese shelf-life using non-starter lactic acid bacteria

Six strains of non-starter lactic acid bacteria (NSLAB) were used to extend the shelf-life of the fresh cheese Tosèla manufactured with pasteurised cows’ milk. The acidification kinetics of three Lactobacillus paracasei, one Lactobacillus rhamnosus and two Streptococcus macedonicus were studied in synthetic milk medium. Lb. paracasei NdP78 and NdP88 and S. macedonicus NdP1 and PB14-1 showed an interesting acidifying capacity and were further characterised for growth in UHT milk and production of antimicrobial compounds. Lb. paracasei NdP78 and S. macedonicus NdP1 grew more than 2 log cycles in 6 h. Lb. paracasei NdP78 was also found to produce a bacteriocin-like inhibitory substance (BLIS) active against Listeria monocytogenes. The four NSLAB strains (singly or in combination) were used to produce experimental pilot-scale cheeses which were compared by a panel. The cheese manufactured with the mixed culture Lb. paracasei NdP78 - S. macedonicus NdP1 was the most appreciated for its sensory properties. The cheeses produced at factory-scale showed higher concentrations of lactobacilli (7.90 log CFU/g) and streptococci (6.10 log CFU/g), but a lower development of coliforms (3.10 log CFU/g) and staphylococci (2.78 log CFU/g) than control cheese (4.86, 4.89, 4.93 and 5.00 log CFU/g of lactobacilli, streptococci, coliforms and staphylococci, respectively) processed without NSLAB addition. The food pathogens Salmonella spp. and Listeria monocytogenes were never detected. The dominance of the species inoculated was demonstrated by denaturing gradient gel electrophoresis (DGGE), whereas strain recognition was evaluated by randomly amplified polymorphic DNA (RAPD)-PCR. From the results obtained, Lb. paracasei NdP78 and S. macedonicus NdP1 were able to persist during the storage of Tosèla cheese and their combination influenced positively the sensory characteristics and shelf-life of the final product.     

Compliant assembly tolerance analysis: guidelines to formalize the resistance spot welding plasticity effects

The variational analysis of compliant assemblies is mainly based on linear elastic models. Some guidelines have been defined to integrate material plasticity into a tolerance analysis model in order to improve its results when considering the resistance spot welding (RSW) process. A finite element model that simulates the body-in-white and RSW processes has been applied to butt and slip joints, with parts subjected to dimensional and geometrical tolerances that cause gap mismatching condition and loading interference on fixtures. The dimensional quality of assemblies is affected by plasticization near the welding spot, at the base of welded flanges and near the locators. The springback evidenced relative rotation of parts.     

Improving System Identification Using Clustering

System identification involves identification of a behavioral model that best explains the measured behavior of a structure. This research uses a strategy of generation and iterative filtering of multiple candidate models for system identification. The task of model filtering is supported by measurement-interpretation cycles. During each cycle, the location for subsequent measurement is chosen using the predictions of current candidate models. In this paper, data mining techniques are proposed to support such measurement-interpretation cycles. Candidate models, representing possible states of a structure, are clustered using a technique that combines principal component analysis and K-means clustering. Representative models of each cluster are used to place sensors for subsequent measurement on the basis of the entropy of their predictions. Results show that clustering is necessary to identify the different groups of candidate models. The entropy of predictions is found to be a valid stopping criterion for iterative sensor addition. Clustering helps classify models and, thus, it provides useful support to engineers for further decision making.     

Model based scenario studies to optimize the regional nitrogen balance and reduce leaching of nitrate and sulfate of an agriculturally used water catchment

Oxidation of pyrite by nitrate (autotrophic denitrification) was identified as the main cause for sulfate increase in drinking water wells in an agriculturally used watershed, located in the north of Lower Saxony (Germany). Nitrate, which inducts this microbial catalyzed process, is drained into ground water predominantly from agricultural fertilization. The increase of sulfate in the ground water can only be stopped by reducing nitrate leaching into the ground water. To analyze the negative influence of agricultural fertilization on the quality of ground water different fertilization strategies were deducted for an investigated area of 890 ha. Calculated on the basis of nutrient balance of soil surface, the current average nitrogen balance in the investigated area amounts to 91 kg N ha^-1 a⁻¹. Farm-gate balance of nutrients is a strong indicator for assessing potential nutrient losses caused by leaching. This indicator shows comparable accuracy to the calculated nutrient balance of soil surface which demands, however, much more data input for calculations. Nitrate concentrations in seepage water in 2 m depth layer of the soil from agricultural fields were simulated with the model HERMES for the whole investigated area (agricultural land + forest). The nitrate concentration in seepage water was calculated for the whole area on the basis of farm-gate nutrient balance as an annual average, which amounts to 14.0 mg NO₃–N l⁻¹ (62 mg NO₃ l⁻¹). In order to keep the nitrate concentration of the ground water below the threshold value for drinking water (EU-water directive: 11.3 mg NO₃–N l⁻¹ (50 mg NO₃ l⁻¹) and to limit pyrite oxidation, different scenarios with simulation studies to optimize fertilization measures were developed. Only those scenarios which assured reduction of an average nitrate concentration in the drainage water below 11.3 mg NO₃–N l⁻¹ (50 mg NO₃ l⁻¹) without profit cuts for the farms were analyzed.

Effects of gamma irradiation on the viability and phenotypic characteristics of Salmonella Enteritidis inoculated into specific-pathogen-free eggs

The goal of this study was to determine the effects of various levels of gamma irradiation on the phenotypic characteristics of 20 strains of Salmonella Enteritidis inoculated separately into specific-pathogen-free shell eggs. Bacterial strains were inoculated into egg yolks and exposed to (60)Co radiation at doses of 0.49 to 5.0 kGy. The eggs were maintained at 25°C and analyzed for the presence of Salmonella on days 1, 2, 4, and 7, and the recovered Salmonella isolates were characterized biochemically. All strains were resistant to doses of 0.49, 0.54, 0.59, 0.8, and 1 kGy; colony counts were ≥10(5) CFU/ml of egg yolk except for one strain, which was detected at 96 h and at 7 days after irradiation at 1 kGy, with a population reduction of 2 log CFU/ml. For the other evaluated doses, 12 strains (60.0%) were resistant at 1.5 kGy and 7 strains (35.0%) were resistant at 3.0 kGy. Among all analyzed strains, 5.0 kGy was more effective for reducing and/or eliminating the inoculated bacteria; only two (10%) strains were resistant to this level of irradiation. Salmonella colony counts were significantly reduced (P < 0.01) with increasing doses from the day 1 to 7 of observation, when microbial growth peaked. Loss of mobility, lactose fermentation, citrate utilization, and hydrogen sulfide production occurred in some strains after irradiation independent of dose and postirradiation storage time. Increases in antibiotic susceptibility also occurred: seven strains became sensitive to β-lactams, two strains became sensitive to antifolates, and one strain each became sensitive to fluoroquinolone, phenicol, nitrofurans, tetracyclines, and aminoglycosides. The results indicate that up to 5.0 kGy of radiation applied to shell eggs inoculated with Salmonella Enteritidis at 4 log CFU per egg is not sufficient for complete elimination of this pathogen from this food matrix.     

Oxidative Stress in Malaria: Potential Benefits of Antioxidant Therapy

Malaria is an infectious disease and a serious public health problem in the world, with 3.3 billion people in endemic areas in 100 countries and about 200 million new cases each year, resulting in almost 1 million deaths in 2018. Although studies look for strategies to eradicate malaria, it is necessary to know more about its pathophysiology to understand the underlying mechanisms involved, particularly the redox balance, to guarantee success in combating this disease. In this review, we addressed the involvement of oxidative stress in malaria and the potential benefits of antioxidant supplementation as an adjuvant antimalarial therapy.     

Disentangling Phase and Morphological Evolution During the Formation of the Lithium Superionic Conductor Li10GeP2S12

The structural and morphological changes of the Lithium superionic conductor Li₁₀GeP₂S₁₂, prepared via a widely used ball milling-heating method over a comprehensive heat treatment range (50 – 700 °C), are investigated. Based on the phase composition, the formation process can be distinctly separated into four zones: Educt, Intermediary, Formation, and Decomposition zone. It is found that instead of Li₄GeS₄–Li₃PS₄ binary crystallization process, diversified intermediate phases, including GeS₂ in different space groups, multiphasic lithium phosphosulfides (Li_xP_yS_z), and cubic Li₇Ge₃PS₁₂ phase, are involved additionally during the formation and decomposition of Li₁₀GeP₂S₁₂. Furthermore, the phase composition at temperatures around the transition temperatures of different formation zones shows a significant deviation. At 600 °C, Li₁₀GeP₂S₁₂ is fully crystalline, while the sample decomposed to complex phases at 650 °C with 30 wt.% impurities, including 20 wt.% amorphous phases. These findings over such a wide temperature range are first reported and may help provide previously lacking insights into the formation and crystallinity control of Li₁₀GeP₂S₁₂.