bluuebunny/crossref_metadata_2025 huggingface dataset:
Drought Superimposes the Positive Effect of Silver Fir on Water Relations of European Beech in Mature Forest Stands
Authors: Magh Ruth-Kristina , Bonn Boris , Grote Rüdiger , Burzlaff Tim , Pfautsch Sebastian , Rennenberg Heinz
Published: October 2019

Research Highlights: Investigations of evapotranspiration in a mature mixed beech-fir forest stand do not indicate higher resilience towards intensified drying-wetting cycles as compared with pure beech stands. Background and Objectives: Forest management seeks to implement adaptive measures, for example, the introduction of more drought resistant species into prevailing monospecific stands to minimize forest mortality and monetary losses. In Central Europe this includes the introduction of native silver fir (Abies alba) into monospecific beech (Fagus sylvatica) stands. In order to determine, if the introduction of fir would improve the resilience against drier conditions, this study investigates water relations of a mature pure beech and a mature mixed beech-fir stand under natural as well as reduced water availability. Materials and Methods: Sap flow rates and densities were measured in two consecutive years using the heat ratio method and scaled using stand inventory data and modeling. Results: Transpiration rates estimated from sap flow were significantly higher for beech trees as compared with silver fir which was attributed to the more anisohydric water-use strategy of the beech trees. We estimate that stand evapotranspiration was slightly higher for mixed stands due to higher interception losses from the mixed stand during times of above average water supply. When precipitation was restricted, beech was not able to support its transpiration demands, and therefore there was reduced sap flow rates in the mixed, as well as in the pure stand, whereas transpiration of fir was largely unaffected, likely due to its more isohydric behavior toward water use and access to moister soil layers. Thus, we found the rates of evapotranspiration in the mixed beech-fir stand to be smaller during times with no precipitation as compared with the pure beech stand, which was accountable to the severely reduced transpiration of beech in the mixed stand. Conclusions: We conclude that smaller evapotranspiration rates in the mixed beech-fir stand might not be the result of increased water use efficiency but rather caused by restricted hydraulic conductivity of the root system of beech, making mixed beech-fir stands at this site less resilient towards drought.
Intra-Annual Radial Growth of Pinus kesiya var. langbianensis Is Mainly Controlled by Moisture Availability in the Ailao Mountains, Southwestern China
Authors: Fan Ze-Xin , Bräuning Achim , Fu Pei-Li , Yang Rao-Qiong , Qi Jin-Hua , Grießinger Jussi , Gebrekirstos Aster
Published: October 2019

Intra-annual monitoring of tree growth dynamics is increasingly applied to disentangle growth-change relationships with local climate conditions. However, such studies are still very limited in subtropical regions which show a wide variety of climate regimes. We monitored stem radius variations (SRV) of Pinus kesiya var. langbianensis (Szemao pine) over five years (2012–2015 and 2017) in the subtropical monsoon mountain climate of the Ailao Mountains, Yunnan Province, southwest China. On average, the stem radial growth of Szemao pine started in early March and ended in early October, and the highest growth rates occurred during May to June. Stem radius increments were synchronous with precipitation events, while tree water deficit corresponded to the drought periods. Correlation analysis and linear mixed-effects models revealed that precipitation and relative humidity are the most important limiting factors of stem radial increments, whereas air temperature and vapor pressure deficit significantly affected tree water balance and may play an important role in determining the growing season length and seasonality (i.e., duration, start, and cessation). This study reveals that moisture availability plays a major role for tree growth of P. kesiya var langbianensis in the Ailao Mountains, southwest China.
The Effect of Sleep Curtailment on Hedonic Responses to Liquid and Solid Food
Authors: Szczygiel Edward J. , Cho Sungeun , Tucker Robin M.
Published: October 2019

It is currently unclear whether changes in sweet taste perception of model systems after sleep curtailment extend to complex food matrices. Therefore, the primary objective of this study was to use a novel solid oat-based food (crisps) and oat-based beverage stimulus sweetened with sucralose to assess changes in taste perception after sleep curtailment. Forty-one participants recorded a habitual and curtailed night of sleep using a single-channel electroencephalograph. The next morning, overall sweetness, flavor, and texture liking responses to energy- and nutrient-matched oat products across five concentrations of sweetness were measured. Overall (p = 0.047) and flavor (p = 0.017) liking slopes across measured concentrations were steeper after curtailment, suggesting that sweeter versions of the oat products were liked more after sleep curtailment. Additionally, a hierarchical cluster analysis was used to classify sweet likers and non-likers. While the effect of sleep curtailment on sweet liking did not differ between sweet liking classification categories, sleep curtailment resulted in decreased texture liking in the solid oat crisps for sweet non-likers (p < 0.001), but not in the oat beverage. These findings illustrate the varied effects of sleep on hedonic response in complex food matrices and possible mechanisms by which insufficient sleep can lead to sensory-moderated increases in energy intake.
Volatile Composition, Sensory Profile and Consumer Acceptability of HydroSOStainable Table Olives
Authors: Sánchez-Rodríguez Lucía , Cano-Lamadrid Marina , Carbonell-Barrachina Ángel A. , Sendra Esther , Hernández Francisca
Published: October 2019

HydroSOStainable table olives (cultivar Manzanilla) are produced from olive trees grown under regulated deficit irrigation (RDI) strategies. Olives produced by RDI are known to have a higher content of some bioactive compounds (e.g. polyphenols), but no information about consumer acceptance (or liking) have been reported so far. In this study, the volatile composition, the sensory profile and the consumer opinion and willingness to pay (at three locations) for HydroSOStainable table olives produced from three RDI treatments and a control were studied. Volatile composition was affected by RDI, by increasing alcohols, ketones and phenolic compounds in some treatments, while others led to a decrease in esters and the content of organic acids. Descriptive sensory analysis (10 panelists) showed an increase of green-olive flavor with a decrease of bitterness in the HydroSOStainable samples. Consumers (study done with 100 consumers in 2-rural and 1-urban locations; ntotal = 300), after being informed about the HydroSOStainable concept, preferred HydroSOStainable table olives to the conventional samples and were willing to pay a higher price for them (52% 1.35–1.75 € and 32% 1.75–2.50 € as compared to the regular price of 1.25 € for a 200 g bag). Finally, green-olive flavor, hardness, crunchiness, bitterness, sweetness and saltiness were defined as the attributes driving consumer acceptance of HydroSOStainable table olives.
The Effect of Processing Methods on Phytochemical Composition in Bergamot Juice
Authors: Cautela Domenico , Vella Filomena Monica , Laratta Bruna
Published: October 2019

Experimental and epidemiological studies show a positive relation between consumption of citrus juices and reduction of risk for some chronic disorders, such as diabetes and cardiovascular diseases. In particular, the bergamot juice is characterized by noticeable amounts of phytochemicals such as flavanone glycosides, limonoids, and quaternary ammonium compounds, all health-beneficial biomolecules. In vitro and in vivo studies have shown anti-inflammatory, cholesterol-lowering, and anti-diabetic activities attributed to these compounds depending on their chemical structure. However, nutritional content of bergamot juice may vary as consequence of different processing techniques, thus needing to address this claim. For this reason, the objective of this research was to evaluate the effects of different processing systems on the proximate constituents, the composition, and the antioxidant activity of the correspondent juices. Overall, the results indicate that the process employed may influence the chemical composition and the functional properties of the ended juice. Screw press method produced a juice with greater content of flavanone glycosides (ranged from 37 to 402 mg/L) and limonoid aglycones (ranged from 65 to 67 mg/L) than the other processes (p < 0.001). However, the process used for extraction of bergamot juice did not affect significantly the N,N-dimethyl-L-proline content (p < 0.5). Moreover, the screw press juice showed the highest antioxidant activity with EC50 value of 9.35 µg/mL, thus suggesting that this method maintains for health the nutritional quality of a fresh-pressed juice.
Occurrence and Residue Concentration of Coccidiostats in Feed and Food of Animal Origin; Human Exposure Assessment
Authors: Roila Rossana , Branciari Raffaella , Pecorelli Ivan , Cristofani Elisa , Carloni Cristiano , Ranucci David , Fioroni Laura
Published: October 2019

Occurring central Italy, 262 unmedicated feed samples and 353 samples of animal tissues and eggs are tested for coccidiostats between 2012 and 2017. A validated multi-residue HPLC-MS/MS method is applied for the simultaneous determination of the 11 coccidiostats licensed in the EU. The dietary exposure to coccidiostats through poultry meat and eggs is calculated for high consumers, and the contribution to acceptable daily intake of coccidiostats is evaluated. The occurrence of positive feed samples ranges from 17.2% in 2012 to 28.3% in 2017, with an average percentage of positive samples of 25%, while 3.8% of feed samples are non-compliant with a concentration ranging from 0.015 mg/kg for diclazuril to 56 mg/kg for narasin. Positive samples of animal tissues, on average, are 34.7%, fully compliant, while 16% of eggs are positive and violative residues are found in 2%. These noncompliant samples show a concentration varying from 2.4 µg/kg to 1002 µg/kg. The contribution of poultry meat and egg consumption to the acceptable daily intake of each coccidiostat is below 1%, highlighting a low direct risk to public health.
The Influence of Steeping Water Change during Malting on the Multi-Toxin Content in Malt
Authors: Habschied Kristina , Krska Rudolf , Sulyok Michael , Lukinac Jasmina , Jukić Marko , Šarkanj Bojan , Krstanović Vinko , Mastanjević Krešimir
Published: October 2019

The aim of this study was to assess the impact of steeping water change and Fusarium graminearum contamination level on different multi-toxin types and concentrations in barley malt. Malt samples were subjected to two micromalting regimes—steeping water change and the other with no steeping water change. Malt was contaminated with different F. graminearum contamination levels (0%, 10%, and 20%). The results indicate that malt with higher F. graminearum contamination levels ensured higher concentrations of toxins. Higher fungal metabolite concentrations were determined in samples exposed to freshly-changed steeping water, especially zearalenone and its derivates whose values were three to four times higher than in samples with no water change. Zearalenone-4-sulfate showed four (in 10% contamination) and even thirty times (in 20% contamination) higher concentrations than in samples with no water change. Water change during malting resulted in higher levels of multi-toxins in the final product.
High-Throughput 16S rRNA Sequencing to Assess Potentially Active Bacteria and Foodborne Pathogens: A Case Example in Ready-to-Eat Food
Authors: Mira Miralles Marina , Maestre-Carballa Lucia , Lluesma-Gomez Monica , Martinez-Garcia Manuel
Published: October 2019

Technologies to detect the entire bacterial diversity spectra and foodborne pathogens in food represent a fundamental advantage in the control of foodborne illness. Here, we applied high-throughput 16S rRNA sequencing of amplicons obtained by PCR and RT-PCR from extracted DNA and RNA targeting the entire bacterial community and the active bacterial fraction present in some of the most consumed and distributed ready-to-eat (RTE) salad brands in Europe. Customer demands for RTE food are increasing worldwide along with the number of associated foodborne illness and outbreaks. The total aerobic bacterial count in the analyzed samples was in the range of 2–4 × 106 CFU/g (SD ± 1.54 × 106). Culture validated methods did not detect Salmonella spp., Escherichia coli, and other fecal coliforms. 16S rRNA gene Illumina next-generation sequencing (NGS) data were congruent with these culture-based results and confirmed that these and other well-known foodborne bacterial pathogens, such as Listeria, were not detected. However, the fine-resolution of the NGS method unveiled the presence of the opportunistic pathogens Aeromonas hydrophyla and Rahnella aquatilis (relative frequency of 1.33–7.33%) that were metabolically active in addition to non-pathogenic, active members of Yersinia spp. (relative frequency of 0.0015–0.003%). The common ail and foxA marker genes of Yersinia enterocolitica were not detected by qPCR. Finally, our NGS data identified to non-pathogenic Pseudomonas spp. as the most abundant and metabolically active bacteria in the analyzed RTE salads (53–75% of bacterial abundance). Our data demonstrate the power of sequencing, in parallel, both 16S rRNA and rDNA to identify and discriminate those potentially and metabolically active bacteria and pathogens to provide a more complete view that facilitates the control of foodborne diseases, although further work should be conducted to determine the sensitivity of this method for targeting bacteria
Risk Management of Dairy Product Losses as a Tool to Improve the Environment and Food Rescue
Authors: Bilska Beata , Kołożyn-Krajewska Danuta
Published: October 2019

“Food loss”, defined as food produced for human consumption, which for various reasons leaves the supply chain, can be assigned to a group of new risks. Irrational use of food constitutes a risk to the environment. Moreover, food losses represent a missed opportunity to improve global food security. The aim of this study was to develop a risk management model for dairy product losses using the example of ripening cheese. The necessary data to develop the model were derived from a survey that was conducted in five dairies located in Poland. Total losses for nine products amounted to 1.1% of the average annual production, which accounted for more than 5635 t per annum. The studies that were conducted allowed the identification of three management methods of food loss in dairies: reprocessing, hand over for feed, and disposal. The level of risk was defined as “high” with two suggested courses of action: prevention and tolerance. Risks must be prevented by eliminating any errors that may result in a product of inadequate quality. Another solution is to redistribute or sell products at a reduced price, which despite their reduced quality, are nevertheless suitable for consumption. To some extent, this risk must be tolerated.
Milk Emulsions: Structure and Stability
Authors: Braun Katja , Hanewald Andreas , Vilgis Thomas A.
Published: October 2019

The main aim of this research is to investigate the characteristics of milk and milk proteins as natural emulsifiers. It is still largely unclear how the two main fractions of the milk proteins behave as emulsifier in highly concentrated emulsions. The surface-active effect of these is determined experimentally for emulsions with a high oil content (φ > 0.7), in this case fully refined rapeseed oil. Recent publications have not yet sufficiently investigated how proteins from native milk behave in emulsions in which a jamming transition is observed. In addition, scientific measurements comparing fresh milk emulsions and emulsions of dried milk protein powders based on rheological and thermal properties are pending and unexamined. The emulsions, prepared with a rotor-stator disperser, are investigated by their particle size and analysed by microscopy, characterised by their rheological properties. The behaviour under shear is directly observed by rheo-optical methods, which enables the direct observation of the dynamic behaviour of the oil droplets undergoing a size selective jamming transition. For a better understanding of the contributions of the different emulsifying proteins, oil-in-water emulsions have been prepared by using whey protein isolates and sodium casinates. Their different role (and function) on the interface activity can be assigned to the droplet sizes and mechanical behaviour during increasing shear deformation. In addition, solid (gelled) emulsions are prepared by heating. It is shown that the cysteine-containing whey proteins are mainly responsible for the sol–gel transition in the continuous water phase and the formation of soft solids.
Human Colorectal Cancer from the Perspective of Mouse Models
Authors: Stastna Monika , Janeckova Lucie , Hrckulak Dusan , Kriz Vitezslav , Korinek Vladimir
Published: October 2019

Colorectal cancer (CRC) is a heterogeneous disease that includes both hereditary and sporadic types of tumors. Tumor initiation and growth is driven by mutational or epigenetic changes that alter the function or expression of multiple genes. The genes predominantly encode components of various intracellular signaling cascades. In this review, we present mouse intestinal cancer models that include alterations in the Wnt, Hippo, p53, epidermal growth factor (EGF), and transforming growth factor β (TGFβ) pathways; models of impaired DNA mismatch repair and chemically induced tumorigenesis are included. Based on their molecular biology characteristics and mutational and epigenetic status, human colorectal carcinomas were divided into four so-called consensus molecular subtype (CMS) groups. It was shown subsequently that the CMS classification system could be applied to various cell lines derived from intestinal tumors and tumor-derived organoids. Although the CMS system facilitates characterization of human CRC, individual mouse models were not assigned to some of the CMS groups. Thus, we also indicate the possible assignment of described animal models to the CMS group. This might be helpful for selection of a suitable mouse strain to study a particular type of CRC.
A New Geodetic Method of Examination of Geometrical Conditions of a Crane Bridge
Authors: Filipiak-Kowszyk Daria , Kamiński Waldemar , Makowska Karolina
Published: October 2019

Safety is one of the key aspects related to crane-based material transport. In order to ensure safe crane operation and material transport, it is necessary to meet certain geometrical conditions. The authors addressed the geometrical conditions of a crane bridge, a substantial crane component. The paper presents the method to compute displacement components of points on the top of a bridge crane relative to their design position. Theoretical considerations presented in the paper have been verified on simulated data. Keeping in mind the proper operation of a crane bridge, the authors proposed in the analyses the use of geometrical relations (perpendicular and parallel character) between end carriages and girders, not previously included in the available literature. The obtained results show that the presented method may be successfully applied to check the geometry of a crane bridge.
Mediating Climate, Mediating Scale
Authors: Pasek Anne
Published: October 2019

Climate communication is seemingly stuck in a double bind. The problem of global warming requires inherently trans-scalar modes of engagement, encompassing times and spaces that exceed local frames of experience and meaning. Climate media must therefore negotiate representational extremes that risk overwhelming their audience with the immensity of the problem or rendering it falsely manageable at a local scale. The task of visualizing climate is thus often torn between scales germane to the problem and scales germane to individuals. In this paper I examine how this scalar divide has been negotiated visually, focusing in particular on Ed Hawkins’ 2016 viral climate spiral. To many, the graphic represents a promising union of political and scientific communication in the public sphere. However, formal analysis of the gif’s reception suggest that the spiral was also a site of anxiety and negative emotion for many viewers. I take these conflicting interpretations as cause to rethink current assumptions about best practices and desirable outcomes for scalar mediations of climate and their capacities to mobilize a wide range of reactions and interpretations—some more legibly political and some more complicatedly affective, yet all nevertheless integral to the work of building a holistic response to the climate crisis.
Effects of Resistant Starch Ingestion on Postprandial Lipemia and Subjective Appetite in Overweight or Obese Subjects
Authors: García-Vázquez Carlos , Ble-Castillo Jorge L. , Arias-Córdova Yolanda , Córdova-Uscanga Rubén , Tovilla-Zárate Carlos A. , Juárez-Rojop Isela E. , Olvera-Hernández Viridiana , Alvarez-Villagomez Carina S. , Nolasco-Coleman Ana M. , Díaz-Zagoya Juan C.
Published: October 2019

Reports surrounding the role of resistant starch (RS) on postprandial lipemia in humans are scarce. The aim of the present study is to examine the effects of resistant starch on the postprandial lipemic response, subjective measures of appetite, and energy intake in overweight and obese subjects. In a randomized, single-blind, crossover study, 14 overweight/obese participants ate a high-fat breakfast (679 kcal, 58% from fat) and a supplement with native banana starch (NBS), high-amylose maize starch (HMS), or digestible maize starch (DMS) on three separate occasions. All supplements provided were matched by the available carbohydrate content, and the RS quantity in NBS and HMS supplements was identical. Appetite was estimated using visual analogue scale (VAS) and an ad libitum test meal. Postprandial glycemia, triglycerides, cholesterol, high-density lipoprotein (HDL) cholesterol, and insulin excursions did not differ between treatments. Subjective appetite measures of satiety were significantly increased after HMS; however, no effects on energy intake were observed during the ad libitum test meal. These findings suggest that a single acute dose of RS cannot be expected to improve postprandial lipemia in subjects with overweight or obesity on a high-fat meal. However, the potential benefits of long-term supplementation should not be ruled out based on these results.
Modelling and Simulation of Selected Real Estate Market Spatial Phenomena
Authors: Kobylińska Katarzyna , Cellmer Radosław
Published: October 2019

This paper presents a novel approach to the modelling and simulation of real estate transactions. The main purpose of the study was to develop the theoretical foundations for building simulation models of transaction locations and real estate prices. Pursuing this objective involved a spatial market analysis based on geostatistics to develop maps of the dynamics and spatial activity of the real estate market. The research was conducted by presenting the issue against the background of the literature of the subject and by conducting an experiment, which involved developing an original procedure of providing simulated market data. The study deals with the market for non-built-up land real estate with a residential function in the city of Olsztyn (Poland). The time range concerned the years 2004–2015. Information on 932 real estate transactions was adopted for the study. A set of additional information on virtual transactions was generated during the study; this information can supplement market data for markets of low activity or if there are information gaps. Geoinformation analyses were performed in order to determine new trends in price levels and spatial activity of a real estate market. Overall, this resulted in generating maps of simulated transaction densities, a map of simulated prices and a map of the probability of a specific price occurring.
Mixed-Generation PAMAM G3-G0 Megamer as a Drug Delivery System for Nimesulide: Antitumor Activity of the Conjugate Against Human Squamous Carcinoma and Glioblastoma Cells
Authors: Zaręba Magdalena , Sareło Przemysław , Kopaczyńska Marta , Białońska Agata , Uram Łukasz , Walczak Małgorzata , Aebisher David , Wołowiec Stanisław
Published: October 2019

Polyhydroxylated dendrimer was synthesized from poly(amidoamine) (PAMAM) dendrimer generation 3 by addition of glycidol (G3gl). G3gl megamer was further modified by binding PAMAM G0 dendrimers by activation of G3gl with p-nitrophenylchloroformate, followed by the addition of excess PAMAM G0 and purification using dialysis. The maximum G0 binding capacity of G3gl was 12 in the case when G0 was equipped with two covalently attached nimesulide equivalents. Nimesulide (N) was converted into N-(p-nitrophenyl) carbonate derivative and fully characterized using X-ray crystallography and spectral methods. Nimesulide was then attached to G0 via a urea bond to yield G02N. The mixed generation G3gl–G02N megamer was characterized using 1H NMR spectroscopy, and its molecular weight was estimated to be 22.4 kDa. The AFM image of G3gl–G02N deposited on mica demonstrated aggregation of nimesulide-covered megamer. The height of the deposited megamer was 8.5 nm. The megameric conjugate with nimesulide was tested in vitro on three human cell lines: squamous cell carcinoma (SCC-15) and glioblastoma (U-118 MG) overexpressing cyclooxygenase-2 (COX-2), and normal skin fibroblasts (BJ). The conjugate efficiently penetrated into all cells and was more cytotoxic against SCC-15 than against BJ. Moreover, the conjugate produced a strong and selective antiproliferative effect on both cancer cell lines (IC50 < 7.5 µM).
Inducible Cardiac-Specific Deletion of Sirt1 in Male Mice Reveals Progressive Cardiac Dysfunction and Sensitization of the Heart to Pressure Overload
Authors: Sanz Maria-Nieves , Grimbert Lucile , Moulin Maryline , Gressette Mélanie , Rucker-Martin Catherine , Lemaire Christophe , Mericskay Mathias , Veksler Vladimir , Ventura-Clapier Renée , Garnier Anne , Piquereau Jérôme
Published: October 2019

Heart failure is associated with profound alterations of energy metabolism thought to play a major role in the progression of this syndrome. SIRT1 is a metabolic sensor of cellular energy and exerts essential functions on energy metabolism, oxidative stress response, apoptosis, or aging. Importantly, SIRT1 deacetylates the peroxisome proliferator-activated receptor gamma co-activator 1α (PGC-1α), the master regulator of energy metabolism involved in mitochondrial biogenesis and fatty acid utilization. However, the exact role of SIRT1 in controlling cardiac energy metabolism is still incompletely understood and conflicting results have been obtained. We generated a cardio-specific inducible model of Sirt1 gene deletion in mice (Sirt1ciKO) to decipher the role of SIRT1 in control conditions and following cardiac stress induced by pressure overload. SIRT1 deficiency induced a progressive cardiac dysfunction, without overt alteration in mitochondrial content or properties. Sixteen weeks after Sirt1 deletion an increase in mitochondrial reactive oxygen species (ROS) production and a higher rate of oxidative damage were observed, suggesting disruption of the ROS production/detoxification balance. Following pressure overload, cardiac dysfunction and alteration in mitochondrial properties were exacerbated in Sirt1ciKO mice. Overall the results demonstrate that SIRT1 plays a cardioprotective role on cardiac energy metabolism and thereby on cardiac function.
Effects of Membrane and Biological Target on the Structural and Allosteric Properties of Recoverin: A Computational Approach
Authors: Borsatto Alberto , Marino Valerio , Abrusci Gianfranco , Lattanzi Gianluca , Dell’Orco Daniele
Published: October 2019

Recoverin (Rec) is a prototypical calcium sensor protein primarily expressed in the vertebrate retina. The binding of two Ca2+ ions to the functional EF-hand motifs induces the extrusion of a myristoyl group that increases the affinity of Rec for the membrane and leads to the formation of a complex with rhodopsin kinase (GRK1). Here, unbiased all-atom molecular dynamics simulations were performed to monitor the spontaneous insertion of the myristoyl group into a model multicomponent biological membrane for both isolated Rec and for its complex with a peptide from the GRK1 target. It was found that the functional membrane anchoring of the myristoyl group is triggered by persistent electrostatic protein-membrane interactions. In particular, salt bridges between Arg43, Arg46 and polar heads of phosphatidylserine lipids are necessary to enhance the myristoyl hydrophobic packing in the Rec-GRK1 assembly. The long-distance communication between Ca2+-binding EF-hands and residues at the interface with GRK1 is significantly influenced by the presence of the membrane, which leads to dramatic changes in the connectivity of amino acids mediating the highest number of persistent interactions (hubs). In conclusion, specific membrane composition and allosteric interactions are both necessary for the correct assembly and dynamics of functional Rec-GRK1 complex.
Resistance Mechanisms to CAR T-Cell Therapy and Overcoming Strategy in B-Cell Hematologic Malignancies
Authors: Song Moo-Kon , Park Byeong-Bae , Uhm Ji-Eun
Published: October 2019

Chimeric antigen receptor (CAR) T-cell therapy has shown promising clinical impact against hematologic malignancies. CD19 is a marker on the surface of normal B cells as well as most B-cell malignancies, and thus has a role as an effective target for CAR T-cell therapy. In numerous clinical data, successes with cell therapy have provided anticancer therapy as a potential therapeutic option for patients who are resistant to standard chemotherapies. However, recent growing evidence showed the limitations of the treatment such as antigen-positive relapse due to poor CAR T-cell persistence and antigen-negative relapses associated with CAR-driven mutations, alternative splicing, epitope masking, low antigen density, and lineage switching. The understanding of the resistance mechanisms to the cell therapy has developed novel potential treatment strategies, including dual-targeting therapy (dual and tandem CAR), and armored and universal CAR T-cell therapies. In this review, we provide an overview of resistance mechanisms to CD19 CAR T-cell therapy in B-cell malignancies and also review therapeutic strategies to overcome these resistances.
Control of Arms of Au Stars Size and its Dependent Cytotoxicity and Photosensitizer Effects in Photothermal Anticancer Therapy
Authors: Depciuch Joanna , Stec Malgorzata , Maximenko Alexey , Pawlyta Miroslawa , Baran Jarek , Parlinska-Wojtan Magdalena
Published: October 2019

Gold nanostars (AuS NPs) are a very attractive nanomaterial, which is characterized by high effective transduction of the electromagnetic radiation into heat energy. Therefore, AuS NPs can be used as photosensitizers in photothermal therapy (PTT). However, understanding the photothermal conversion efficiency in nanostars is very important to select the most appropriate shape and size of AuS NPs. Therefore, in this article, the synthesis of AuS NPs with different lengths of star arms for potential application in PTT was investigated. Moreover, the formation mechanism of these AuS NPs depending on the reducer concentration is proposed. Transmission electron microscopy (TEM) with selected area diffraction (SEAD) and X-ray diffraction (X-Ray) showed that all the obtained AuS NPs are crystalline and have cores with similar values of the diagonal (parameter d), from 140 nm to 146 nm. However, the widths of the star arm edges (parameter c) and the lengths of the arms (parameter a) vary between 3.75 nm and 193 nm for AuS1 NPs to 6.25 nm and 356 nm for AuS4 NPs. Ultraviolet-visible (UV-Vis) spectra revealed that, with increasing edge widths and lengths of the star arms, the surface plasmon resonance (SPR) peak is shifted to the higher wavelengths, from 640 nm for AuS1 NPs to 770 nm for AuS4 NPs. Moreover, the increase of temperature in the AuS NPs solutions as well as the values of calculated photothermal efficiency grew with the elongation of the star arms. The potential application of AuS NPs in the PTT showed that the highest decrease of viability, around 75%, of cells cultured with AuS NPs and irradiated by lasers was noticed for AuS4 NPs with the longest arms, while the smallest changes were visible for gold nanostars with the shortest arms. The present study shows that photothermal properties of AuS NPs depend on edge widths and lengths of the star arms and the values of photothermal efficiency are higher with the increase of the arm lengths, which is correlated with the reducer concentration.
Translesion DNA Synthesis Across Lesions Induced by Oxidative Products of Pyrimidines: An Insight into the Mechanism by Microscale Thermophoresis
Authors: Hrabina Ondrej , Brabec Viktor , Novakova Olga
Published: October 2019

Oxidative stress in cells can lead to the accumulation of reactive oxygen species and oxidation of DNA precursors. Oxidized nucleotides such as 2’-deoxyribo-5-hydroxyuridin (HdU) and 2’-deoxyribo-5-hydroxymethyluridin (HMdU) can be inserted into DNA during replication and repair. HdU and HMdU have attracted particular interest because they have different effects on damaged-DNA processing enzymes that control the downstream effects of the lesions. Herein, we studied the chemically simulated translesion DNA synthesis (TLS) across the lesions formed by HdU or HMdU using microscale thermophoresis (MST). The thermodynamic changes associated with replication across HdU or HMdU show that the HdU paired with the mismatched deoxyribonucleoside triphosphates disturbs DNA duplexes considerably less than thymidine (dT) or HMdU. Moreover, we also demonstrate that TLS by DNA polymerases across the lesion derived from HdU was markedly less extensive and potentially more mutagenic than that across the lesion formed by HMdU. Thus, DNA polymerization by DNA polymerase η (polη), the exonuclease-deficient Klenow fragment of DNA polymerase I (KF–), and reverse transcriptase from human immunodeficiency virus type 1 (HIV-1 RT) across these pyrimidine lesions correlated with the different stabilization effects of the HdU and HMdU in DNA duplexes revealed by MST. The equilibrium thermodynamic data obtained by MST can explain the influence of the thermodynamic alterations on the ability of DNA polymerases to bypass lesions induced by oxidative products of pyrimidines. The results also highlighted the usefulness of MST in evaluating the impact of oxidative products of pyrimidines on the processing of these lesions by damaged DNA processing enzymes.
Future Preventive Gene Therapy of Polygenic Diseases from a Population Genetics Perspective
Authors: Oliynyk Roman Teo
Published: October 2019

With the accumulation of scientific knowledge of the genetic causes of common diseases and continuous advancement of gene-editing technologies, gene therapies to prevent polygenic diseases may soon become possible. This study endeavored to assess population genetics consequences of such therapies. Computer simulations were used to evaluate the heterogeneity in causal alleles for polygenic diseases that could exist among geographically distinct populations. The results show that although heterogeneity would not be easily detectable by epidemiological studies following population admixture, even significant heterogeneity would not impede the outcomes of preventive gene therapies. Preventive gene therapies designed to correct causal alleles to a naturally-occurring neutral state of nucleotides would lower the prevalence of polygenic early- to middle-age-onset diseases in proportion to the decreased population relative risk attributable to the edited alleles. The outcome would manifest differently for late-onset diseases, for which the therapies would result in a delayed disease onset and decreased lifetime risk; however, the lifetime risk would increase again with prolonging population life expectancy, which is a likely consequence of such therapies. If the preventive heritable gene therapies were to be applied on a large scale, the decreasing frequency of risk alleles in populations would reduce the disease risk or delay the age of onset, even with a fraction of the population receiving such therapies. With ongoing population admixture, all groups would benefit over generations.
Comparing the Osteogenic Potentials and Bone Regeneration Capacities of Bone Marrow and Dental Pulp Mesenchymal Stem Cells in a Rabbit Calvarial Bone Defect Model
Authors: Lee Yu-Chieh , Chan Ya-Hui , Hsieh Sung-Chih , Lew Wei-Zhen , Feng Sheng-Wei
Published: October 2019

The bone regeneration efficiency of bone marrow mesenchymal stem cells (BMSCs) and dental pulp mesenchymal stem cells (DPSCs) combined with xenografts in the craniofacial region remains unclear. Accordingly, this study commenced by comparing the cell morphology, cell proliferation, trilineage differentiation, mineral synthesis, and osteogenic gene expression of BMSCs and DPSCs in vitro. Four experimental groups (empty control, Bio-Oss only, Bio-Oss+BMSCs, and Bio-Oss+DPSCs) were then designed and implanted in rabbit calvarial defects. The BMSCs and DPSCs showed a similar morphology, proliferative ability, surface marker profile, and trilineage-differentiation potential in vitro. However, the BMSCs exhibited a higher mineral deposition and expression levels of osteogenic marker genes, including alkaline phosphatase (ALP), runt related transcription factor 2 (RUNX2), and osteocalcin (OCN). In the in vivo studies, the bone volume density in both MSC groups was significantly greater than that in the empty control or Bio-Oss only group. Moreover, the new bone formation and Collagen I / osteoprotegerin protein expressions of the scaffold+MSC groups were higher than those of the Bio-Oss only group. Finally, the Bio-Oss+BMSC and Bio-Oss+DPSC groups had a similar bone mineral density, new bone formation, and osteogenesis-related protein expression. Overall, the DPSCs seeded on Bio-Oss matched the bone regeneration efficacy of BMSCs in vivo and hence appear to be a promising strategy for craniofacial defect repair in future clinical applications.
Predicting Gonadal Germ Cell Cancer in People with Disorders of Sex Development; Insights from Developmental Biology
Authors: Looijenga Leendert H. J. , Kao Chia-Sui , Idrees Muhammad T.
Published: October 2019

The risk of gonadal germ cell cancer (GGCC) is increased in selective subgroups, amongst others, defined patients with disorders of sex development (DSD). The increased risk is due to the presence of part of the Y chromosome, i.e., GonadoBlastoma on Y chromosome GBY region, as well as anatomical localization and degree of testicularization and maturation of the gonad. The latter specifically relates to the germ cells present being at risk when blocked in an embryonic stage of development. GGCC originates from either germ cell neoplasia in situ (testicular environment) or gonadoblastoma (ovarian-like environment). These precursors are characterized by presence of the markers OCT3/4 (POU5F1), SOX17, NANOG, as well as TSPY, and cKIT and its ligand KITLG. One of the aims is to stratify individuals with an increased risk based on other parameters than histological investigation of a gonadal biopsy. These might include evaluation of defined susceptibility alleles, as identified by Genome Wide Association Studies, and detailed evaluation of the molecular mechanism underlying the DSD in the individual patient, combined with DNA, mRNA, and microRNA profiling of liquid biopsies. This review will discuss the current opportunities as well as limitations of available knowledge in the context of predicting the risk of GGCC in individual patients.
Directed Evolution of an Improved Rubisco; In Vitro Analyses to Decipher Fact from Fiction
Authors: Zhou Yu , Whitney Spencer
Published: October 2019

Inaccuracies in biochemically characterizing the amount and CO2-fixing properties of the photosynthetic enzyme Ribulose-1,5-bisphosphate (RuBP) carboxylase/oxygenase continue to hamper an accurate evaluation of Rubisco mutants selected by directed evolution. Here, we outline an analytical pipeline for accurately quantifying Rubisco content and kinetics that averts the misinterpretation of directed evolution outcomes. Our study utilizes a new T7-promoter regulated Rubisco Dependent Escherichia coli (RDE3) screen to successfully select for the first Rhodobacter sphaeroides Rubisco (RsRubisco) mutant with improved CO2-fixing properties. The RsRubisco contains four amino acid substitutions in the large subunit (RbcL) and an improved carboxylation rate (kcatC, up 27%), carboxylation efficiency (kcatC/Km for CO2, increased 17%), unchanged CO2/O2 specificity and a 40% lower holoenzyme biogenesis capacity. Biochemical analysis of RsRubisco chimers coding one to three of the altered amino acids showed Lys-83-Gln and Arg-252-Leu substitutions (plant RbcL numbering) together, but not independently, impaired holoenzyme (L8S8) assembly. An N-terminal Val-11-Ile substitution did not affect RsRubisco catalysis or assembly, while a Tyr-345-Phe mutation alone conferred the improved kinetics without an effect on RsRubisco production. This study confirms the feasibility of improving Rubisco by directed evolution using an analytical pipeline that can identify false positives and reliably discriminate carboxylation enhancing amino acids changes from those influencing Rubisco biogenesis (solubility).
Beneficial Effects of Neurotensin in Murine Model of Hapten-Induced Asthma
Authors: Russjan Ewelina , Kaczyńska Katarzyna
Published: October 2019

Neurotensin (NT) demonstrates ambiguous activity on inflammatory processes. The present study was undertaken to test the potential anti-inflammatory activity of NT in a murine model of non-atopic asthma and to establish the contribution of NTR1 receptors. Asthma was induced in BALB/c mice by skin sensitization with dinitrofluorobenzene followed by intratracheal hapten provocation. The mice were treated intraperitoneally with NT, SR 142948 (NTR1 receptor antagonist) + NT or NaCl. Twenty-four hours after the challenge, airway responsiveness to nebulized methacholine was measured. Bronchoalveolar lavage fluid (BALF) and lungs were collected for biochemical and immunohistological analysis. NT alleviated airway hyperreactivity and reduced the number of inflammatory cells in BALF. These beneficial effects were inhibited by pretreatment with the NTR1 antagonist. Additionally, NT reduced levels of IL-13 and TNF-α in BALF and IL-17A, IL12p40, RANTES, mouse mast cell protease and malondialdehyde in lung homogenates. SR 142948 reverted only a post-NT TNF-α decrease. NT exhibited anti-inflammatory activity in the hapten-induced asthma. Reduced leukocyte accumulation and airway hyperresponsiveness indicate that this beneficial NT action is mediated through NTR1 receptors. A lack of effect by the NTR1 blockade on mast cell activation, oxidative stress marker and pro-inflammatory cytokine production suggests that other pathways can be involved, which requires further research.
Suitability of GnRH Receptors for Targeted Photodynamic Therapy in Head and Neck Cancers
Authors: Pethő Lilla , Murányi József , Pénzes Kinga , Gurbi Bianka , Brauswetter Diána , Halmos Gábor , Csík Gabriella , Mező Gábor
Published: October 2019

Head and neck squamous cell carcinomas (HNSCC) have a high mortality rate, although several potential therapeutic targets have already been identified. Gonadotropin-releasing hormone receptor (GnRH-R) expression is less studied in head and neck cancers, hence, we investigated the therapeutic relevance of GnRH-R targeting in HNSCC patients. Our results indicate that half of the patient-derived samples showed high GnRH-R expression, which was associated with worse prognosis, making this receptor a promising target for GnRH-based drug delivery. Photodynamic therapy is a clinically approved treatment for HNSCC, and the efficacy and selectivity may be enhanced by the covalent conjugation of the photosensitizer to a GnRH-R targeting peptide. Several native ligands, gonadotropin-releasing hormone (GnRH) isoforms, are known to target GnRH-R effectively. Therefore, different 4Lys(Bu) modified GnRH analogs were designed and conjugated to protoporphyrin IX. The receptor binding potency of the novel conjugates was measured on human pituitary and human prostate cancer cells, indicating only slightly lower GnRH-R affinity than the peptides. The in vitro cell viability inhibition was tested on Detroit-562 human pharyngeal carcinoma cells that express GnRH-R in high levels, and the results showed that all conjugates were more effective than the free protoporphyrin IX.
Effects of Omega-3 Fatty Acids on Immune Cells
Authors: Gutiérrez Saray , Svahn Sara L , Johansson Maria E
Published: October 2019

Alterations on the immune system caused by omega-3 fatty acids have been described for 30 years. This family of polyunsaturated fatty acids exerts major alterations on the activation of cells from both the innate and the adaptive immune system, although the mechanisms for such regulation are diverse. First, as a constitutive part of the cellular membrane, omega-3 fatty acids can regulate cellular membrane properties, such as membrane fluidity or complex assembly in lipid rafts. In recent years, however, a new role for omega-3 fatty acids and their derivatives as signaling molecules has emerged. In this review, we describe the latest findings describing the effects of omega-3 fatty acids on different cells from the immune system and their possible molecular mechanisms.
Ceramide and Sphingosine Regulation of Myelinogenesis: Targeting Serine Palmitoyltransferase Using microRNA in Multiple Sclerosis
Authors: Dasgupta Somsankar , Ray Swapan K.
Published: October 2019

Ceramide and sphingosine display a unique profile during brain development, indicating their critical role in myelinogenesis. Employing advanced technology such as gas chromatography–mass spectrometry, high performance liquid chromatography, and immunocytochemistry, along with cell culture and molecular biology, we have found an accumulation of sphingosine in brain tissues of patients with multiple sclerosis (MS) and in the spinal cord of rats induced with experimental autoimmune encephalomyelitis. The elevated sphingosine leads to oligodendrocyte death and fosters demyelination. Ceramide elevation by serine palmitoyltransferse (SPT) activation was the primary source of the sphingosine elevation as myriocin, an inhibitor of SPT, prevented sphingosine elevation and protected oligodendrocytes. Supporting this view, fingolimod, a drug used for MS therapy, reduced ceramide generation, thus offering partial protection to oligodendrocytes. Sphingolipid synthesis and degradation in normal development is regulated by a series of microRNAs (miRNAs), and hence, accumulation of sphingosine in MS may be prevented by employing miRNA technology. This review will discuss the current knowledge of ceramide and sphingosine metabolism (synthesis and breakdown), and how their biosynthesis can be regulated by miRNA, which can be used as a therapeutic approach for MS.