Evaluation of a Novel Plasmid for Simultaneous Gene Electrotransfer-Mediated Silencing of CD105 and CD146 in Combination with Irradiation

Targeting tumor vasculature through specific endothelial cell markers represents a promising approach for cancer treatment. Here our aim was to construct an antibiotic resistance gene-free plasmid encoding shRNAs to simultaneously target two endothelial cell markers, CD105 and CD146, and to test its functionality and therapeutic potential in vitro when delivered by gene electrotransfer (GET) and combined with irradiation (IR). Functionality of the plasmid was evaluated by determining the silencing of the targeted genes using qRT-PCR. Antiproliferative and antiangiogenic effects were determined by the cytotoxicity assay tube formation assay and wound healing assay in murine endothelial cells 2H-11. The functionality of the plasmid construct was also evaluated in malignant melanoma tumor cell line B16F10. Additionally, potential activation of immune response was measured by induction of DNA sensor STING and proinflammatory cytokines by qRT-PCR in endothelial cells 2H-11. We demonstrated that the plasmid construction was successful and can efficiently silence the expression of the two targeted genes. As a consequence of silencing, reduced migration rate and angiogenic potential was confirmed in 2H-11 endothelial cells. Furthermore, induction of DNA sensor STING and proinflammatory cytokines were determined, which could add to the therapeutic effectiveness when used in vivo. To conclude, we successfully constructed a novel plasmid DNA with two shRNAs, which holds a great promise for further in vivo testing.     

Agronomic performance of high oleic,low linolenic soybean in Tennessee

Soybean oil hydrogenation alters the linolenic acid molecule to prevent the oil from becoming rancid, however, health reports have indicated trans-fat caused by hydrogenation, is not generally regarded as safe. Typical soybeans contain approximately 80 g kg⁻¹ to 120 g kg⁻¹ linolenic acid and 240 g kg⁻¹ of oleic acid. In an effort to accommodate the need for high-quality oil, the United Soybean Board introduced an industry standard for a high oleic acid greater than 750 g kg⁻¹ and linolenic acid less than 30 g kg⁻¹ oil. By combing mutations in the soybean plant at four loci, FAD2-1A and FAD2-1B, oleate desaturase genes and FAD3A and FAD3C, linoleate desaturase genes, and seed oil will not require hydrogenation to prevent oxidation and produce high-quality oil. In 2017 and 2018, a study comparing four near-isogenic lines across multiple Tennessee locations was performed to identify agronomic traits associated with mutations in FAD3A and FAD3C loci, while holding FAD2-1A and FAD2-1B constant in the mutant (high oleic) state. Soybean lines were assessed for yield and oil quality based on mutations at FAD2-1 and FAD3 loci. Variations of wild-type and mutant genotypes were compared at FAD3A and FAD3C loci. Analysis using a generalized linear mixed model in SAS 9.4, indicated no yield drag or other negative agronomic traits associated with the high oleic and low linolenic acid genotype. All four mutations of fad2-1A, fad2-1B, fad3A, and fad3C were determined as necessary to produce a soybean with the new industry standard (>750 g kg⁻¹ oleic and <30 g kg⁻¹ linolenic acid) in a maturity group-IV-Late cultivar for Tennessee growers.     

Depassivation and repassivation of stainless steels by stepwise pH change

Immersion tests with different stainless steels have been performed, while the pH was stepwise decreased and then increased again. During 8.5-day exposure, the depassivation and repassivation pH values as a function of pitting resistance equivalent number were determined. There is always a gap between both pH values (depassivation and repassivation), indicating that for every steel, there are conditions where an existing passive layer can be maintained but cannot be rebuilt after depassivation. In such environments, the passive layer is thicker, consisting mainly of molybdenum and iron rich oxides, while chromium is dissolved. Usually, depending on conditions, the passive layer is more chromium-rich, especially the inner layer. This is relevant, for example, for acidizing jobs in oil and gas industry, proving that repassivation after acidizing will happen promptly, when the pH is increased again.     

Field Performance of High Oleic Soybeans with Mutant FAD2-1A and FAD2-1B Genes in Tennessee

Soybean [Glycine max (L.) Merr.] oil with high oleic acid (>75%) has increased oxidative stability and health benefits that are valuable for food, fuel, and industrial products. It has been determined that two naturally occurring mutations in genes FAD2-1A and FAD2-1B can combine to produce high oleic soybeans. The objective of this study was to test the effect of these mutant alleles on seed yield and oil and protein concentration. Molecular markers assisted in the creation of a population of 48 BC₃F_2:4 lines (93.75% expected genome commonality). Each line was classified into one of four genotypic groups where both FAD2-1A and FAD2-1B genes were either homozygous wild type or mutant, respectively. Twelve lines for each genotypic group were evaluated in three replications at six locations across Tennessee. There was no seed yield difference between the high oleic genotypic group and the other groups (P < 0.05). On the other hand, there were differences in fatty acid profiles and oil and protein concentrations. In combination, the mutant FAD2-1A and FAD2-1B alleles produced a mean of 803.1 g kg⁻¹ oleic acid. This is, on average, approximately 500 g kg⁻¹ more oleic acid compared to soybean lines with only one mutant FAD2-1 allele. The high oleic double mutant group had more total oil (228.0 g kg⁻¹) and protein (401.0 g kg⁻¹) compared to all other genotypic groups (P < 0.05). Overall, this specific combination of mutant FAD2-1A and FAD2-1B alleles appears to generate conventional high oleic soybeans without a yield drag.     

NaWuReT-Workshop: Forschung in der Reaktionstechnik für und mit der Gesellschaft

In this article, the results of the NaWuReT (Early Career Reaction Engineers) workshop on the topic “Are we doing relevant science?” are presented. The topics “(In)surmountable hurdles for Citizen Scientists in reaction engineering?” and “Circular Economy in reaction engineering with/for society?” were discussed. Therefrom, a variety of ideas and suggestions were extracted.     

Impact of adaptive comfort criteria and heat waves on optimal building thermal mass control

This article investigates building thermal mass control of commercial buildings to reduce utility costs with a particular emphasis on the individual impacts of both adaptive comfort criteria and of heat waves. Recent changes in international standards on thermal comfort for indoor environments allow for adaptation to the weather development as manifested in comfort criteria prEN 15251.2005 and NPR-CR 1752.2005 relative to the non-adaptive comfort criterion ISO 7730.2003. Furthermore, since extreme weather patterns tend to occur more frequently, even in moderate climate zones, it is of interest how a building's passive thermal storage inventory responds to prolonged heat waves. The individual and compounded effects of adaptive comfort criteria and heat waves on the conventional and optimal operation of a prototypical office building are investigated for the particularly hot month of August 2003 in Freiburg, Germany. It is found that operating commercial buildings using adaptive comfort criteria strongly reduces total cooling loads and associated building systems energy consumption under conventional and building thermal mass control. In the case of conventional control, total operating cost reductions follow the cooling loads reductions closely. Conversely, the use of adaptive comfort criteria under optimal building thermal mass control leads to both lower and slightly higher absolute operating costs compared to the optimal costs for the non-adaptive ISO 7730. While heat waves strongly affect the peak cooling loads under both conventional and optimal building thermal mass control, total cooling loads, building energy consumption and costs are only weakly affected for both control modes. Passive cooling under cost-optimal control, while achieving significant total cost reductions of up to 13%, is associated with total energy penalties on the order of 1–3% relative to conventional nighttime setup control. Thus, building thermal mass control defends its cost saving potential under optimal control in the presence of adaptive comfort criteria and heat waves.