Radiolysis of Amino Acids by Heavy and Energetic Cosmic Ray Analogs in Simulated Space Environments: α\alpha-Glycine Zwitterion Form

In this work, we studied the stability of the glycine molecule in the crystalline zwitterion form, known as {\alpha}-glycine ($^{+}$NH$_{3}$CH$_{2}$COO$^{-}$) under action of heavy cosmic ray analogs. The experiments were conducted in a high vacuum chamber at heavy ions accelerator GANIL, in Caen, France. The samples were bombarded at two temperatures (14 K and 300 K) by $^{58}$Ni$^{11+}$ ions of 46 MeV until the final fluence of $10^{13}$ ions cm$^{-2}$. The chemical evolution of the sample was evaluated in-situ using Fourrier Transformed Infrared (FTIR) spectrometer. The bombardment at 14 K produced several daughter species such as OCN$^-$, CO, CO$_2$, and CN$^-$. The results also suggest the appearing of peptide bonds during irradiation but this must be confirmed by further experiments. The halflives of glycine in Interstellar Medium were estimated to be 7.8 $\times 10^3$ years (300 K) and 2.8 $\times 10^3$ years (14 K). In the Solar System the values were 8.4 $\times 10^2$ years (300 K) and 3.6 $\times 10^3$ years (14 K). It is believed that glycine could be present in space environments that suffered aqueous changes such as the interior of comets, meteorites and planetesimals. This molecule is present in proteins of all alive beings. So, studying its stability in these environments provides further understanding about the role of this specie in the prebiotic chemistry on Earth.Comment: 28 pages, 12 figures, 9 tables. Accepted to be published at Monthly Notices of the Royal Astronomical Society (MNRAS

Methods for generating year-round access to amphioxus in the laboratory.

Cephalochordates, commonly known as amphioxus, are key to understanding vertebrate origins. However, laboratory work suffers from limited access to adults and embryonic material. Here we report the design and experimental validation of an inland marine facility that allows establishing stable amphioxus colonies in the laboratory and obtaining embryos at any time of day and over almost the entire year, far exceeding natural conditions. This is achieved by mimicking the natural benthic environment, natural day- and moon- light, natural substrate and by providing a strictly controlled and seasonally fluctuating temperature regimen. Moreover, supplemented algae diets allow animals to refill their gonads in consecutive years. Spontaneous spawning, a major problem in previous setups, no longer occurs in our facility; instead, all breeding is induced and fertilization occurs fully in vitro. Our system makes amphioxus a standard laboratory animal model

CRISPR-Mediated VHL Knockout Generates an Improved Model for Metastatic Renal Cell Carcinoma.

Metastatic renal cell carcinoma (mRCC) is nearly incurable and accounts for most of the mortality associated with RCC. Von Hippel Lindau (VHL) is a tumour suppressor that is lost in the majority of clear cell RCC (ccRCC) cases. Its role in regulating hypoxia-inducible factors-1α (HIF-1α) and -2α (HIF-2α) is well-studied. Recent work has demonstrated that VHL knock down induces an epithelial-mesenchymal transition (EMT) phenotype. In this study we showed that a CRISPR/Cas9-mediated knock out of VHL in the RENCA model leads to morphologic and molecular changes indicative of EMT, which in turn drives increased metastasis to the lungs. RENCA cells deficient in HIF-1α failed to undergo EMT changes upon VHL knockout. RNA-seq revealed several HIF-1α-regulated genes that are upregulated in our VHL knockout cells and whose overexpression signifies an aggressive form of ccRCC in the cancer genome atlas (TCGA) database. Independent validation in a new clinical dataset confirms the upregulation of these genes in ccRCC samples compared to adjacent normal tissue. Our findings indicate that loss of VHL could be driving tumour cell dissemination through stabilization of HIF-1α in RCC. A better understanding of the mechanisms involved in this phenomenon can guide the search for more effective treatments to combat mRCC

Parallel Force Assay for Protein-Protein Interactions

Quantitative proteome research is greatly promoted by high-resolution parallel format assays. A characterization of protein complexes based on binding forces offers an unparalleled dynamic range and allows for the effective discrimination of non-specific interactions. Here we present a DNA-based Molecular Force Assay to quantify protein-protein interactions, namely the bond between different variants of GFP and GFP-binding nanobodies. We present different strategies to adjust the maximum sensitivity window of the assay by influencing the binding strength of the DNA reference duplexes. The binding of the nanobody Enhancer to the different GFP constructs is compared at high sensitivity of the assay. Whereas the binding strength to wild type and enhanced GFP are equal within experimental error, stronger binding to superfolder GFP is observed. This difference in binding strength is attributed to alterations in the amino acids that form contacts according to the crystal structure of the initial wild type GFP-Enhancer complex. Moreover, we outline the potential for large-scale parallelization of the assay

Mitteltiefe geothermische Anlagen in Sachsen

Ziel des Forschungsvorhabens war es, die Einsatzpotentiale mitteltiefer geothermischer Systeme für die sächsischen Ballungsräume Leipzig, Dresden und Chemnitz detailliert zu untersuchen, Kostenvergleiche durchzuführen und Handlungsempfehlungen für potenzielle Projekte abzuleiten. Für jeden Musterstandort wurde eine mitteltiefe Erdwärmesondenanlage vollständig geplant. Mitteltiefe Erdwärmeanlagen können demzufolge eine wirtschaftliche Alternative zur konventionellen Klimatisierung/Temperierung von Gebäuden darstellen. Im vorliegenden Forschungsbericht sind Schritte für Planung, bauliche Umsetzung und Überwachung zusammengefasst. Auch Investitions- und Betriebskosten werden abgeschätzt. Damit können die Ergebnisse des Forschungsvorhabens als Entscheidungshilfe für Behörden, Investoren und Fachplaner herangezogen werden. Redaktionsschluss: 31.08.202

Integrated Surface-Groundwater Modelling of Nitrate Concentration in Mediterranean Rivers, the Júcar River Basin District, Spain

[EN] High nutrient discharge from groundwater (GW) into surface water (SW) have multiple undesirable effects on river water quality. With the aim to estimate the impact of anthropic pressures and river-aquifer interactions on nitrate status in SW, this study integrates two hydrological simulation and water quality models. PATRICAL models SW-GW interactions and RREA models streamflow changes due to human activity. The models were applied to the Jucar River Basin District (RBD), where 33% of the aquifers have a concentration above 50 mg NO3-/L. As a result, there is a direct linear correlation between the nitrate concentration in rivers and aquifers (Jucar r(2) = 0.9, and Turia r(2) = 0.8), since in these Mediterranean basins, the main amount of river flows comes from groundwater discharge. The concentration of nitrates in rivers and GW tends to increase downstream of the district, where artificial surfaces and agriculture are concentrated. The total NO3- load to Jucar RBD rivers was estimated at 10,202 tN/year (239 kg/km(2)/year), from which 99% is generated by diffuse pollution, and 3378 tN/year (79 kg/km(2)/year) is discharged into the Mediterranean Sea. Changes in nitrate concentration in the RBD rivers are strongly related to the source of irrigation water, river-aquifer interactions, and flow regulation. The models used in this paper allow the identification of pollution sources, the forecasting of nitrate concentration in surface and groundwater, and the evaluation of the efficiency of measures to prevent water degradation, among other applications.The first author's research is partially funded by a PhD scholarship from the food research stream of the program "Colombia Cientifica-Pasaporte a la Ciencia", granted by the Colombian Institute for Educational Technical Studies Abroad (Instituto Colombiano de Credito Educativo y Estudios Tecnicos en el Exterior, ICETEX). The authors thank the Spanish Research Agency (AEI) for the financial support to RESPHIRA project (PID2019-106322RB-100)/AEI/10.13039/501100011033.Dorado-Guerra, DY.; Paredes Arquiola, J.; Pérez-Martín, MÁ.; Tafur Hermann, H. (2021). Integrated Surface-Groundwater Modelling of Nitrate Concentration in Mediterranean Rivers, the Júcar River Basin District, Spain. Sustainability. 13(22):1-21. https://doi.org/10.3390/su132212835S121132

Parallel Force Assay for Protein-Protein Interactions

Quantitative proteome research is greatly promoted by high-resolution parallel format assays. A characterization of protein complexes based on binding forces offers an unparalleled dynamic range and allows for the effective discrimination of non-specific interactions. Here we present a DNA-based Molecular Force Assay to quantify protein-protein interactions, namely the bond between different variants of GFP and GFP-binding nanobodies. We present different strategies to adjust the maximum sensitivity window of the assay by influencing the binding strength of the DNA reference duplexes. The binding of the nanobody Enhancer to the different GFP constructs is compared at high sensitivity of the assay. Whereas the binding strength to wild type and enhanced GFP are equal within experimental error, stronger binding to superfolder GFP is observed. This difference in binding strength is attributed to alterations in the amino acids that form contacts according to the crystal structure of the initial wild type GFP-Enhancer complex. Moreover, we outline the potential for large-scale parallelization of the assay

Identification of candidate genes for congenital splay leg in piglets by alternative analysis of DNA microarray data

The congenital splay leg syndrome in piglets is characterized by a temporarily impaired functionality of the hind leg muscles immediately after birth. Etiology and pathogenetic mechanisms for the disease are still not well understood. We compared genome wide gene expression of three hind leg muscles (M. adductores, M. gracilis and M. sartorius) between affected piglets and their healthy littermates with the GeneChip® Porcine Genome Array (Affymetrix) in order to identify candidate genes for the disease. Data analysis with standard algorithms revealed no significant differences between both groups. By application of an alternative approach, we identified 63 transcripts with differences in two muscles and 5 genes differing between the groups in three muscles. The expression of six selected genes (SQSTM1, SSRP1, DDIT4, ENAH, MAF, and PDK4) was investigated with SYBRGreen RT - Real time PCR. The differences obtained with the microarray analysis could be confirmed and demonstrate the validity of the alternative approach to microarray data analysis. Four genes with different expression levels in at least two muscles (SQSTM1, SSRP1, DDIT4, and MAF) are assigned to transcriptional cascades related to cell death and may thus indicate pathways for further investigations on congenital splay leg in piglets