Raman spectroscopy as a tool for characterisation of quality parameters in Norwegian grown apples during ripening

This study shows for the first time the feasibility of Raman spectroscopy as a non-destructive method to follow the ripening process of apple fruits. Two different varieties of apples were studied: ‘Aroma’ and ‘Elstar’. By visual inspection, Raman spectra showed that the starch content was higher in ‘Elstar’ apples compared to ‘Aroma’. The degradation of starch over time could be detected in the Raman spectra, indicating that the method can be used to monitor the ripening process. The ripeness markers starch index, soluble solids content (SSC), and the sugars glucose, fructose and sucrose were determined with traditional destructive methods. Cross validated calibration models based on Raman spectroscopy were obtained for all quality parameters, and test set validation offered good results, with R2 in the range 0.4–0.86 for ‘Aroma’ and 0.4–0.95 for ‘Elstar’, respectively. The regression coefficients showed that the calibrations relied on Raman bands associated with starch and different sugars. The results suggest that Raman spectroscopy in the future could be used to determine the optimal time of harvesting and to sort apples into different degrees of ripeness.publishedVersio

Tidewater glaciers as “climate refugia” for zooplankton-dependent food web in Kongsfjorden, Svalbard

With climate warming, many tidewater glaciers are retreating. Fresh, sediment-rich sub-glacial meltwater is discharged at the glacier grounding line, where it mixes with deep marine water resulting in an upwelling of a plume visible in front of the glacial wall. Zooplankton may suffer increased mortality within the plume due to osmotic shock when brought in contact with the rising meltwater. The constant replenishment of zooplankton and juvenile fish to the surface areas attracts surface-foraging seabirds. Because access to other feeding areas, such as the marginal ice zone, has become energetically costly due to reduced sea-ice extent, glacial plumes may become increasingly important as “climate refugia” providing enhanced prey availability. Here, we investigated zooplankton concentrations within the plume and adjacent waters of four tidewater glaciers in Kongsfjorden, Svalbard, in early August 2016 and late July 2017. Our aim was to compare the zooplankton composition, abundance, and isotopic signatures within the plumes to those in adjacent fjord and shelf waters. Our hypothesis was that the plumes resulted in increased zooplankton mortality through osmotic shock and increased prey availability to predators. The mortality due to osmotic shock in the glacial plume was low (<5% dead organisms in samples), although slightly higher than in surrounding waters. This indicates that plumes are inefficient “death traps” for zooplankton. However, the high abundance and biomass of zooplankton within plume areas suggest that the “elevator effect” of rising glacial water supplies zooplankton to the sea surface, thereby enhancing prey availability for surface-feeding seabirds. Thus, our study provides evidence that glacial plumes are important as “climate refugia” for foraging seabirds. Stable isotope signatures showed that the glacial bay zooplankton and fish community represent a distinct isotopic niche. Additionally, zooplankton mortality associated with the plume estimated over 100-days of melt season supports a flux of 12.8 tonnes of organic carbon to benthic communities in the glacial bays. Benthic scavengers, such as Onisimus caricus and Anonyx nugax, were abundant in the glacial bay, where they feed on sinking organic matter.publishedVersio

KappaMask: AI-Based Cloudmask Processor for Sentinel-2

The Copernicus Sentinel-2 mission operated by the European Space Agency (ESA) provides comprehensive and continuous multi-spectral observations of all the Earth's land surface since mid-2015. Clouds and cloud shadows significantly decrease the usability of optical satellite data, especially in agricultural applications; therefore, an accurate and reliable cloud mask is mandatory for effective EO optical data exploitation. During the last few years, image segmentation techniques have developed rapidly with the exploitation of neural network capabilities. With this perspective, the KappaMask processor using U-Net architecture was developed with the ability to generate a classification mask over northern latitudes into the following classes: clear, cloud shadow, semi-transparent cloud (thin clouds), cloud and invalid. For training, a Sentinel-2 dataset covering the Northern European terrestrial area was labelled. KappaMask provides a 10 m classification mask for Sentinel-2 Level-2A (L2A) and Level-1C (L1C) products. The total dice coefficient on the test dataset, which was not seen by the model at any stage, was 80% for KappaMask L2A and 76% for KappaMask L1C for clear, cloud shadow, semi-transparent and cloud classes. A comparison with rule-based cloud mask methods was then performed on the same test dataset, where Sen2Cor reached 59% dice coefficient for clear, cloud shadow, semi-transparent and cloud classes, Fmask reached 61% for clear, cloud shadow and cloud classes and Maja reached 51% for clear and cloud classes. The closest machine learning open-source cloud classification mask, S2cloudless, had a 63% dice coefficient providing only cloud and clear classes, while KappaMask L2A, with a more complex classification schema, outperformed S2cloudless by 17%

Massive right atrial myxoma presenting as syncope and exertional dyspnea: case report

Primary heart neoplasms are rare occurring with an estimated incidence of 0.0017-0.19%. Myxoma is the most prevalent primary heart tumor. The right atrium is an unusual localization, occurring only in 15-20% of myxoma cases. We report a rare case of a massive right atrial myxoma causing tricuspid valve obstruction and presenting as syncope and exertional dyspnea. This case illustrates the influence of myxoma's size, position and mobility as well as patient's body posture and respiration to the development of signs and symptoms. Three-dimensional echocardiography proved useful in surgery planning, allowing a better definition of the tumor outline and attachment

Seasonal plankton dynamics in Kongsfjorden during two years of contrasting environmental conditions

Seasonal plankton time-series data are presented from Kongsfjorden from two years with contrasting environmental conditions. Kongsfjorden (west coast of Spitsbergen – 79°N) integrates inputs from Atlantic and Arctic waters, and glacier run-off, and is thus a prime location to study impacts on ecosystem dynamics of key environmental drivers that are relevant across the Arctic. Despite extensive research in Kongsfjorden, seasonally-resolved data are scarce. From late April/early May to early September 2019 and 2020, we conducted pelagic sampling at a mid-fjord station at mostly weekly to bi-weekly resolution investigating the environmental drivers of phyto- and zooplankton community composition and phenology. During spring 2019, Atlantic water masses with temperatures > 1 °C were found throughout the upper 250 m of the water column, and little sea ice occurred in the fjord. Spring 2020, in turn, was characterized by the presence of local water masses with sub-zero temperatures and relatively extensive sea-ice cover. The most striking contrast between the two years was the difference in phytoplankton spring bloom composition. In 2019, the spring bloom was dominated by the colonial stage of the haptophyte Phaeocystis pouchetii and diatoms played a minor role, while the spring bloom in 2020 was dominated by diatoms of the genus Thalassiosira succeeded by P. pouchetii. Selective grazing by large copepods and water mass structure seem to have been the decisive factors explaining the marked difference in diatom spring bloom biomass between the years while similar spring abundances of P. pouchetii in both years indicated that this species was less impacted by those factors. Our data suggest that differences in spring bloom composition impacted trophic transfer and carbon export. Recruitment of the dominant copepods Calanus finmarchicus and C. glacialis, Cirripedia and euphausiid larvae as well as the export of carbon to the seabed was more efficient during the diatom-dominated compared to the P. pouchetii–dominated spring bloom. In summer, the plankton composition shifted towards a flagellate-dominated community characterized by mixo- and heterotrophic taxa adapted to a lower nutrient regime and strong top-down control by copepod grazers. However, residual silicic acid after the P. pouchetii–dominated spring bloom fueled a late summer diatom bloom in 2019. Our data provide a first glimpse into the environmental drivers of plankton phenology and underline that high-resolution monitoring over many annual cycles is required to resolve the ephemeral variations of plankton populations against the backdrop of climate change.publishedVersio

Essential functions of the 32 kDa subunit of yeast replication protein A

Replication protein A (RPA) is a heterotrimeric (70, 32 and 14 kDa subunits), single-stranded DNA-binding protein required for cellular DNA metabolism. All subunits of RPA are essential for life, but the specific functions of the 32 and 14 kDa subunits remains unknown. The 32 kDa subunit (RPA2) has multiple domains, but only the central DNA-binding domain (called DBD D) is essential for life in Saccharomyces cerevisiae. To define the essential function(s) of RPA2 in S. cerevisiae, a series of site-directed mutant forms of DBD D were generated. These mutant constructs were then characterized in vitro and in vivo. The mutations had minimal effects on the overall structure and activity of the RPA complex. However, several mutants were shown to disrupt crosslinking of RPA2 to DNA and to dramatically lower the DNA-binding affinity of a RPA2-containing subcomplex. When introduced into S. cerevisiae, all DBD D mutants were viable and supported normal growth rates and DNA replication. These findings indicate that RPA2–DNA interactions are not essential for viability and growth in S. cerevisiae. We conclude that DNA-binding activity of RPA2 is dispensable in yeast and that the essential function of DBD D is intra- and/or inter-protein interactions

Human replication protein A unfolds telomeric G-quadruplexes

G-quadruplex structures inhibit telomerase activity and must be disrupted for telomere elongation during S phase. It has been suggested that the replication protein A (RPA) could unwind and maintain single-stranded DNA in a state amenable to the binding of telomeric components. We show here that under near-physiological in vitro conditions, human RPA is able to bind and unfold G-quadruplex structures formed from a 21mer human telomeric sequence. Analyses by native gel electrophoresis, cross-linking and fluorescence resonance energy transfer indicate the formation of both 1:1 and 2:1 complexes in which G-quadruplexes are unfolded. In addition, quadruplex opening by hRPA is much faster than observed with the complementary DNA, demonstrating that this protein efficiently unfolds G-quartets. A two-step mechanism accounting for the binding of hRPA to G-quadruplexes is proposed. These data point to the involvement of hRPA in regulation of telomere maintenance

Biological Insights into the Expression of Translation Initiation Factors from Recombinant CHOK1SV Cell Lines and their Relationship to Enhanced Productivity

Translation initiation is on the critical pathway for the production of monoclonal antibodies (mAb) by mammalian cells. Formation of a closed loop structure comprised of mRNA, a number of eukaryotic initiation factors and ribosomal proteins has been proposed to aid re-initiation of translation and therefore increase global translational efficiency. We have determined mRNA and protein levels of the key components of the closed loop; eukaryotic initiation factors (eIF3a, eIF3b, eIF3c, eIF3h, eIF3i and eIF4G1), poly(A) binding protein (PABP) 1 and PABP interacting protein 1 (PAIP1) across a panel of 30 recombinant mAb-producing GS-CHOK1SV cell lines with a broad range of growth characteristics and production levels of a model recombinant mAb. We have used a multi-level statistical approach to investigate the relationship between key performance indicators (cell growth and recombinant antibody productivity) and the intracellular amounts of target translation initiation factor proteins and the mRNAs encoding them. We show that high-producing cell lines maintain amounts of the translation initiation factors involved in the formation of the closed loop mRNA, maintaining these proteins at appropriate levels to deliver enhanced recombinant protein production. We then utilise knowledge of the amounts of these factors to build predictive models for, and use cluster analysis to identify, high-producing cell lines. This study therefore defines the translation initiation factor amounts that are associated with highly productive recombinant GS-CHOK1SV cell lines that may be targets for screening highly productive cell lines or to engineer new host cell lines with the potential for enhanced recombinant antibody productivity

Metabolomic Profiling Reveals Mitochondrial-Derived Lipid Biomarkers That Drive Obesity-Associated Inflammation

Obesity has reached epidemic proportions worldwide. Several animal models of obesity exist, but studies are lacking that compare traditional lard-based high fat diets (HFD) to “Cafeteria diets" (CAF) consisting of nutrient poor human junk food. Our previous work demonstrated the rapid and severe obesogenic and inflammatory consequences of CAF compared to HFD including rapid weight gain, markers of Metabolic Syndrome, multi-tissue lipid accumulation, and dramatic inflammation. To identify potential mediators of CAF-induced obesity and Metabolic Syndrome, we used metabolomic analysis to profile serum, muscle, and white adipose from rats fed CAF, HFD, or standard control diets. Principle component analysis identified elevations in clusters of fatty acids and acylcarnitines. These increases in metabolites were associated with systemic mitochondrial dysfunction that paralleled weight gain, physiologic measures of Metabolic Syndrome, and tissue inflammation in CAF-fed rats. Spearman pairwise correlations between metabolites, physiologic, and histologic findings revealed strong correlations between elevated markers of inflammation in CAF-fed animals, measured as crown like structures in adipose, and specifically the pro-inflammatory saturated fatty acids and oxidation intermediates laurate and lauroyl carnitine. Treatment of bone marrow-derived macrophages with lauroyl carnitine polarized macrophages towards the M1 pro-inflammatory phenotype through downregulation of AMPK and secretion of pro-inflammatory cytokines. Results presented herein demonstrate that compared to a traditional HFD model, the CAF diet provides a robust model for diet-induced human obesity, which models Metabolic Syndrome-related mitochondrial dysfunction in serum, muscle, and adipose, along with pro-inflammatory metabolite alterations. These data also suggest that modifying the availability or metabolism of saturated fatty acids may limit the inflammation associated with obesity leading to Metabolic Syndrome