Functional Inactivation of Drosophila GCK Orthologs Causes Genomic Instability and Oxidative Stress in a Fly Model of MODY-2

Maturity-onset diabetes of the young (MODY) type 2 is caused by heterozygous inactivating mutations in the gene encoding glucokinase (GCK), a pivotal enzyme for glucose homeostasis. In the pancreas GCK regulates insulin secretion, while in the liver it promotes glucose utilization and storage. We showed that silencing the Drosophila GCK orthologs Hex-A and Hex-C results in a MODY-2-like hyperglycemia. Targeted knock-down revealed that Hex-A is expressed in insulin producing cells (IPCs) whereas Hex-C is specifically expressed in the fat body. We showed that Hex-A is essential for insulin secretion and it is required for Hex-C expression. Reduced levels of either Hex-A or Hex-C resulted in chromosome aberrations (CABs), together with an increased production of advanced glycation end-products (AGEs) and reactive oxygen species (ROS). This result suggests that CABs, in GCK depleted cells, are likely due to hyperglycemia, which produces oxidative stress through AGE metabolism. In agreement with this hypothesis, treating GCK-depleted larvae with the antioxidant vitamin B6 rescued CABs, whereas the treatment with a B6 inhibitor enhanced genomic instability. Although MODY-2 rarely produces complications, our data revealed the possibility that MODY-2 impacts genome integrity.     

The Age-Sensitive Efficacy of Calorie Restriction on Mitochondrial Biogenesis and mtDNA Damage in Rat Liver

Calorie restriction (CR) is the most efficacious treatment to delay the onset of age-related changes such as mitochondrial dysfunction. However, the sensitivity of mitochondrial markers to CR and the age-related boundaries of CR efficacy are not fully elucidated. We used liver samples from ad libitum-fed (AL) rats divided in: 18-month-old (AL-18), 28-month-old (AL-28), and 32-month-old (AL-32) groups, and from CR-treated (CR) 28-month-old (CR-28) and 32-month-old (CR-32) counterparts to assay the effect of CR on several mitochondrial markers. The age-related decreases in citrate synthase activity, in TFAM, MFN2, and DRP1 protein amounts and in the mtDNA content in the AL-28 group were prevented in CR-28 counterparts. Accordingly, CR reduced oxidative mtDNA damage assessed through the incidence of oxidized purines at specific mtDNA regions in CR-28 animals. These findings support the anti-aging effect of CR up to 28 months. Conversely, the protein amounts of LonP1, Cyt c, OGG1, and APE1 and the 4.8 Kb mtDNA deletion content were not affected in CR-28 rats. The absence of significant differences between the AL-32 values and the CR-32 counterparts suggests an age-related boundary of CR efficacy at this age. However, this only partially curtails the CR benefits in counteracting the generalized aging decline and the related mitochondrial involvement.     

Coupling Interrupted Fischer and Multicomponent Joullié-Ugi to Chase Chemical Diversity: from Batch to Sustainable Flow Synthesis of Peptidomimetics

The generation of peptidomimetic substructures for medicinal chemistry purposes requires effective and divergent synthetic methods. We present in this work an efficient flow process that allows quick modulation of reagents for Joullié-Ugi multicomponent reaction, using spiroindolenines as core motifs. This sterically hindered imine equivalent could successfully be diversified using various isocyanides and amino acids in generally good space-time yields. A telescoped flow process combining interrupted Fischer reaction for spiroindolenine synthesis and subsequent Joullié-Ugi-type modification resulted in product formation in very good overall yield in less than 2 hours compared to 48 hours required in batch mode. The developed protocol can be seen as a general tool for rapid and facile generation of peptidomimetic compounds. We also showcase preliminary biological assessments for the prepared compounds.     

Testing EGFR with Idylla on Cytological Specimens of Lung Cancer: A Review

The current standard of care for advanced non-small-cell lung cancer is based on detecting actionable mutations that can benefit from targeted therapy. Comprehensive genetic tests can have long turn-around times, and because EGFR mutations are the most prevalent actionable mutation, a quick detection would enable a prompt initiation of targeted therapy. Furthermore, the scarcity of diagnostic material means that sometimes only cytologic material is available. The Idylla™ EGFR assay is a real-time PCR–based method able to detect 51 EGFR mutations in 2.5 h. Idylla is validated for use only on FFPE sections, but some researchers described their experiences with cytological material. We reviewed the relevant literature, finding four articles describing 471 cases and many types of cytological input material: smears, cell-block sections, suspensions, and extracted DNA. The sensitivity, specificity, and limit of detection appear comparable to those obtained with histological input material, with one exception: the usage of scraped stained smears as input may reduce the accuracy of the test. In conclusion, usage of cytological material as input to the Idylla EGFR test is possible. A workflow where common mutations are tested first and fast, leaving rarer mutations for subsequent comprehensive profiling, seems the most effective approach.     

Characterisation of the Dynamic Interactions between Complex N-Glycans and Human CD22

CD22 (Siglec-2) is a B-cell surface inhibitory protein capable of selectively recognising sialylated glycans, thus dampening autoimmune responses against self-antigens. Here we have characterised the dynamic recognition of complex-type N-glycans by human CD22 by means of orthogonal approaches including NMR spectroscopy, computational methods and biophysical assays. We provide new molecular insights into the binding mode of sialoglycans in complex with h-CD22, highlighting the role of the sialic acid galactose moieties in the recognition process, elucidating the conformational behaviour of complex-type N-glycans bound to Siglec-2 and dissecting the formation of CD22 homo-oligomers on the B-cell surface. Our results could enable the development of additional therapeutics capable of modulating the activity of h-CD22 in autoimmune diseases and malignancies derived from B-cells.     

Vacuum impregnation of chitosan‐based edible coating in minimally processed pumpkin

The goal of this research was to evaluate the use of vacuum impregnation (VI) and soaking techniques (ST) in the application of edible coatings of chitosan and chitosan + lauric acid to minimally processed pumpkins (MPP). The vacuum impregnation method led to greater component incorporation (5.9% and 1.75%, respectively) in the pumpkins when compared to soaking and consequently the formation of more uniform, thicker coatings (25.6 and 22.3 μm, respectively). However, VI caused greater changes in pH, acidity, colour and firmness. Relating to water content and carotenoid content, noncoated pumpkins presented greater losses during the storage period, regardless of impregnation method. The pumpkins with edible coatings, regardless of method, presented lower numbers of psychrotrophic micro‐organisms and coliforms during the storage period. Therefore, soaking was considered the best method for the application of chitosan‐based edible coatings to minimally processed pumpkins, as it led to smaller changes in the properties of the product.     

CO2 sequestration by magnesite mineralisation through interaction of Mg-brine and CO2: integrated laboratory experiments and computerised geochemical modelling

ABSTRACT

Several Carbon Capture and Storage (CCS) techniques have been studied including injections of carbon dioxide (CO₂) into the mature and/or depleted hydrocarbon reservoirs and deep saline aquifers. This work aims to test storing CO₂ into the magnesium-rich evaporite strata and also into the stratigraphic intervals containing Mg-rich brines. The test simulates Mg-carbonation of the synthetic solution obtained from the Mg-evaporite mineral, bischofite – both experimentally in the laboratory condition and also through computerised geochemical simulation. The laboratory experiments, which resulted in the crystallisation of anhydrous magnesite, were analysed. The TOUGHREACT^TM, Geochemist’s Workbench^TM (GWB) and PHREEQC^TM software simulated the experiments as computerised geochemical model and tested the results for natural geological conditions. The geochemical simulations successfully demonstrate the immense CCS potential for the Mg-evaporite (as well as the sedimentary strata charged with Mg-evaporitic brine) at their subsurface geological occurrences at elevated pressure-temperature and high salinity.     

Tribocorrosion and corrosion behavior of stainless steel coated with DLC films in ethanol with different concentrations of water

There has been a recent increase in both the production and consumption of ethanol due to the numerous environmental advantages that it offers, such as the fact that it can be produced from a variety of renewable materials, for instance corn and cellulose, or it can be obtained from sugarcane bagasse and biomass (2nd and 3rd generation ethanol). The result of this is that nowadays ethanol is widely seen as the dominant biofuel – or as a blend component in gasoline or pure fuel - in many countries.However, one disadvantage of the use of ethanol is the high corrosive behavior that occurs when its hygroscopic properties are exposed to a large number of materials. Xiaoyuan Lou and Preet Singh showed that the increase of water concentration in ethanol induces pitting and metal loss. Diamond-Like Carbon (DLC) films may be a solution to this problem due to the fact that they can be deposited inside tubes, offer good protection levels against corrosion, and reduce the friction coefficient and wear.This paper shows the tribocorrosion and corrosion studies of DLC films deposited on stainless steel grade 304 (SS304) substrates in order to gauge its appropriateness usage in the construction of pipelines and fuel storage tanks. The surface morphology was analyzed before and after 14 days of immersion. The tribocorrosion, friction coefficient, and wear rate were studied in ethanol to see the effects of water concentration. The films showed good adherence to the substrates. Corrosion and tribocorrosion results showed that for bare Stainless Steel 304 the increase of the water content increases the corrosion and the friction coefficient. DLC coated samples presented few points of delamination, and the friction coefficient and open circuit potentials were very low compared with the bare sample which was water concentration independent.     

Antioxidant and antimicrobial potential of cajazeira leaves (Spondias mombin) extracts

Cajazeira leaves (Spondias mombin) have their highlighted use as antioxidant and natural antimicrobial, which justifies the objective of this work to evaluate the biological activities of different extracts. In order to evaluate the antioxidant and antimicrobial potentials of the cajazeira leaves, extractions at low pressure and high pressure were performed. The low pressure extractions (PLE) were carried out using Soxhlet (SOX) and tip ultrasound, using different solvents. Supercritical fluid extraction (SFE) was evaluated at temperature of 40?60°C and pressure of 150?300 bar besides extraction with cosolvent. Higher yields were obtained with the use of more polar solvents at LPE. The extracts obtained by SOX with ethanol and others polar solvents presented the best TPC values and antioxidant activity. The extracts at LPE with hexane and ethyl acetate and SFE presented better antimicrobial activity. Through liquid chromatography of high efficiency, it was possible to identify compounds with recognized biological activity, like ellagic acid, gallic acid and catechin.