bluuebunny/crossref_metadata_2025 huggingface dataset:
Age of Laying Hens Significantly Influences the Content of Nutritionally Vital Lipophilic Compounds in Eggs
Authors: Ko Eun-Young , Saini Ramesh Kumar , Keum Young-Soo , An Byoung-Ki
Published: December 2020

This study aimed to comparatively analyze the carotenoids, tocopherols, fatty acids, and sterols (e.g., cholesterol) in the eggs of laying hens of early (24 weeks), intermediate (42 weeks), and late age (74 weeks), utilizing high-performance liquid chromatography (HPLC), gas chromatography (GC), and GC–mass spectrometry (MS). The results revealed the significantly (p < 0.05) highest content of nutritionally vital compounds (per g of egg yolk, fresh weight), which included (all-E-)-lutein (21.8 µg), (all-E-)-zeaxanthin (13.4 µg), α-tocopherol (76.5 µg), oleic acid (C18:1n9c; 83.3 mg), α-linolenic acid (C18:3n3; 0.68 mg), γ-linolenic acid (C18:3n6; 0.47 mg), arachidonic acid (C20:4n6; 8.11 mg), docosahexaenoic acid (DHA; C22:6n3; 2.06 mg), and total monounsaturated fatty acids (MUFAs; 94.7 mg) and n-3 polyunsaturated fatty acids (PUFAs; 2.74 mg) in the eggs of early-age laying hens compared to intermediate and late-age. Surprisingly, cholesterol was not significantly different in the eggs obtained from the different age groups. In contrast, the fat quality indices, including the lowest atherogenic index (AI) and thrombogenic index (TI) values and high hypocholesterolemic/hypercholesterolemic (h/H) fatty acid ratio, indicated the health-beneficial potential associated with fat intake from the eggs of intermediate and late-age laying hens. Overall, the results of the present investigation suggest that eggs from early-age laying hens can be recommended for a higher intake of antioxidant carotenoids and tocopherols, while the eggs from intermediate and late-age hens can be recommended for the intake of good quality fats.
A Critical Appraisal of the Evidence Supporting Consumer Motivations for Alternative Proteins
Authors: Tso Rachel , Lim Amanda JiaYing , Forde Ciarán G.
Published: December 2020

Alternative proteins are receiving increased global attention. This burgeoning interest in plants (especially plant-based meat alternatives), insects, algae, and cultured meat has been attributed to their reported health benefits, lower environmental impact and improved animal welfare compared to conventional animal-based meat. Food producers and the media are promoting acceptance of these products, claiming superior nutritional, environmental and ethical credentials and a desirable novel sensory experience. However, the evidence supporting these claims remains unclear. In this review, we summarise the main evidence underlying the nutritional, sensorial, economical, ethical, and environmental reasons reported for the rise in consumer demand for alternative proteins. We found many of these reasons to lack a strong evidence base. For instance, evidence is emerging for the nutritional benefits of plant-based meat alternatives, but present claims are largely based on established evidence for plant-based diets. Significant research gaps remain, especially longitudinal evidence on the sustained effects of replacing conventional animal-based proteins with alternative sources. For many alternative proteins, challenges exist in achieving desirable sensory properties akin to animal-based meat to promote their acceptance by consumers. Overall, fundamental shifts in the food system are required to create a culture in which healthful and sustainable food choices are the norm.
A Modified Kwee–Van Woerden Method for Eclipse Minimum Timing with Reliable Error Estimates
Authors: Deeg Hans J.
Published: December 2020

The Kwee–van Woerden (KvW) method used for the determination of eclipse minimum times has been a staple in eclipsing binary research for decades, due its simplicity and the independence of external input parameters, which also makes it well-suited to obtaining timings of exoplanet transits. However, its estimates of the timing error have been known to have a low reliability. During the analysis of very precise photometry of CM Draconis eclipses from TESS space mission data, KvW’s original equation for the timing error estimate produced numerical errors, which evidenced a fundamental problem in this equation. This contribution introduces an improved approach for calculating the timing error with the KvW method. A code that implements this improved method, together with several further updates of the original method, are presented. An example of the application to CM Draconis light curves from TESS is given. The eclipse minimum times are derived with the KvW method’s three original light curve folds, but also with five and seven folds. The use of five or more folds produces minimum timings with a substantially better precision. The improved method of error calculation delivers consistent timing errors which are in excellent agreement with error estimates obtained by other means. In the case of TESS data from CM Draconis, minimum times with an average precision of 1.1 s are obtained. Reliable timing errors are also a valuable indicator for evaluating if a given scatter in an O-C diagram is caused by measurement errors or by a physical period variation.
Development of Case Numbers during the COVID-19 Pandemic in a Center of Maximum-Care for Traumatology and Orthopedic Oncology
Authors: Polan Christina , Burggraf Manuel , Kauther Max Daniel , Meyer Heinz-Lothar , Rademacher Friederike , Braitsch Henrik , Jöckel Karl-Heinz , Hardes Jendrik , Streitbürger Arne , Dudda Marcel
Published: December 2020

(1) Background: The COVID-19 pandemic has led to a significant change in the utilization of trauma surgery and tumor orthopedic hospital facilities. (2) Methods: In a monocentric retrospective analysis, the weekly numbers of cases requiring intra-clinical treatment in the first four months of 2020 were compared with those of 2019. Patients’ visits to the emergency department and shock room, consultation hours, work-related accidents, case numbers in the normal and intensive care units, ventilation hours, the “Simplified Acute Physiology Score/ Therapeutic Intervention Scoring System” (SAPS/TISS), the average length of stay in hospital, the number of operations and their degree of urgency, as well as deaths, were analyzed in a study based on the data from 7606 outpatient consultations in 2019 and 6755 in 2020, as well as 993 inpatient cases in 2019 and 950 in 2020. (3) Results: There was a significant reduction in the number of treatments per week in the emergency department (261 ± 29 vs. 165 ± 25; p < 0.001) with the same number of shock room treatments and fewer consultation hour contacts (226 ± 29 vs. 119 ± 65; p = 0.012). There were fewer inpatient cases (66 ± 7 vs. 42 ± 11; p = 0.001), resulting in a fall in the days of hospitalization (492 ± 63 vs. 308 ± 78; p < 0.001) and number of operations (73 ± 7 vs. 55 ± 10; p = 0.012), especially elective procedures (20 ± 3 vs. 7 ± 7; p = 0.008). The SAPS/TISS score was lower (1351 ± 1213 vs. 399 ± 281; p = 0.023). Fewer fracture treatments and septic surgeries were performed, while the number of procedures to treat orthopedic malignancies remained constant. (4) Conclusions: During the first phase of the COVID-19 pandemic, we observed a significant reduction in the number of cases treated in orthopedics. While the number of multiple-injured patients was unchanged, fewer patients presented for primary and regular care. Treatment of acute injuries and malignant tumor diseases was not at risk. There was no effect on in-house mortality. We see a potential for the recruitment of medical staff from the outpatient department, operating room, and the ward. In the event of a future second wave, our results may allow for early planning, particularly of the all-important human resources. Reorganization by hospitals and decreased patient numbers in trauma surgery can enable the reallocation of medical staff, equipment, and beds to increase capacity for COVID-19 patients.
Numerical and Experimental Investigation of Rectangular Liquid-Containing Structures under Seismic Excitation
Authors: Bahreini Toussi Iman , Kianoush Reza , Mohammadian Abdolmajid
Published: December 2020

Liquid-containing structures are used for various municipal and industrial applications. The functionality of these structures in seismic regions is crucial. The main purpose of this study is to investigate the behavior of liquid under seismic excitations using numerical modelling. For this purpose, experimental and numerical studies are conducted. In the experimental tests, a ground-supported rectangular tank is excited on a shaking table. The tests are videotaped from two directions and subsequently analyzed frame-by-frame. Four different orientations are tested to investigate the effect of bilateral excitation. In the numerical simulations, the same tank is modeled in OpenFOAM—a computational fluid dynamics program—and the same excitations are applied. The results from the numerical and the experimental studies are compared, and reliability of the numerical model is discussed. Furthermore, using the numerical model, the pressure on the roof of the tank is obtained at various locations and examined for different excitations.
Deterioration of the Mechanical Properties of FFF 3D-Printed PLA Structures
Authors: Yonezawa Asahi , Yamada Akira
Published: December 2020

Poly(lactic acid) (PLA) is a biodegradable polymer material used for the fabrication of objects by fused filament fabrication (FFF) 3D printing. FFF 3D printing technology has been quickly spreading over the past few years. An FFF-3D-printed object is formed from melted polymer extruded from a nozzle layer-by-layer. The mechanical properties of the object, and the changes in those properties as the object degrades, differ from the properties and changes observed in bulk objects. In this study we evaluated FFF-3D-printed objects by uniaxial tensile tests and four-point flexural tests to characterize the changes of three mechanical properties, namely, the maximum stress, elastic modulus, and breaking energy. Eight types of test pieces printed directly by an FFF 3D printer using two scan patterns and two interior fill percentages (IFPs) were tested by the aforesaid methods. The test pieces were immersed in saline and kept in an incubator at 37 °C for 30, 60, or 90 days before the mechanical testing. The changes in the mechanical properties differed largely between the test piece types. In some of the test pieces, transient increases in strength were observed before the immersion degraded the strength. Several of the test piece types were found to have superior specific strength in the tests. The results obtained in this research will be helpful for the design of PLA structures fabricated by FFF 3D printing.
Plasma Amino Acid Response to Whey Protein Ingestion Following 28 Days of Probiotic (Bacillus subtilis DE111) Supplementation in Active Men and Women
Authors: Townsend Jeremy R. , Vantrease William C. , Jones Megan D. , Sapp Philip A. , Johnson Kent D. , Beuning Cheryle N. , Haase Allison A. , Boot Claudia M.
Published: December 2020

We sought to determine if 28 days of probiotic supplementation influenced the plasma amino acid (AA) response to acute whey protein feeding. METHODS: Twenty-two recreationally active men (n = 11; 24.3 ± 3.2 yrs; 89.3 ± 7.2 kg) and women (n = 11; 23.0 ± 2.8 yrs; 70.2 ± 15.2 kg) participated in this double-blind, placebo-controlled, randomized study. Before (PRE) and after 28 days of supplementation (POST), participants reported to the lab following a 10-hr fast and provided a resting blood draw (0 min), then subsequently consumed 25 g of whey protein. Blood samples were collected at 15-min intervals for 2 h post-consumption (15–120 min) and later analyzed for plasma leucine, branched-chain AA (BCAA), essential AA (EAA), and total AA (TAA). Participants received a probiotic (PROB) consisting of 1 x10-9 colony forming units (CFU) Bacillus subtilis DE111 (n = 11) or a maltodextrin placebo (PL) (n = 11) for 28 days. Plasma AA response and area under the curve (AUC) values were analyzed via repeated measures analysis of variance. RESULTS: Our analysis indicated no significant (p < 0.05) differential responses for plasma leucine, BCAA, EAA, or TAA between PROB and PL from PRE to POST. AUC analysis revealed no group × time interaction for plasma leucine (p = 0.524), BCAA (p = 0.345), EAA (p = 0.512), and TAA (p = 0.712). CONCLUSION: These data indicate that 28 days of Bacillus subtilis DE111 does not affect plasma AA appearance following acute whey protein ingestion.
Correlation between Underwater Noise and Sea Level at Ieodo Ocean Research Station
Authors: Kim Seong Hyeon , Choi Bok Kyoung , Kim Byoung-Nam
Published: December 2020

To investigate the correlation between underwater noise and sea level, data collected at Ieodo Ocean Research Station (from 15 May to 27 August 2013) were analyzed. Temporal variations in the overall level of underwater noise were similar to those in sea level. The average underwater noise levels at frequencies below 50 Hz were 99.0–106.6 dB during spring tides and 78.4–90.2 dB during neap tides, with a difference of 12.6–20.6 dB. When low-pass filtering at six hours was applied to the absolute value of sea level, a high correlation coefficient of 0.7 for its association with underwater noise was obtained. This strong correlation could indicate flow noise caused by currents. Underwater noise levels around the frequency of 16 Hz were relatively strongly correlated with sea level (correlation coefficient of 0.71). Wavelet analysis of the overall level of underwater noise showed strong periodicity at 0.25, 0.50, 1.0, 16.0, and 30.0 days, as well as characteristics of a tidal constituent. In wavelet coherence analysis, the overall level at a period of 0.25 days exhibited a phase difference of −90 degrees relative to sea level. This finding indicates that the noise level peaks at 1.5 h before the high tide or low tide, which was confirmed in the correlation analysis of noise level and sea height with various time lags. This study shows that the overall level of underwater noise from ebb currents is higher than that from flood currents due to differences in current velocity.
Priming as a Diagnostic of Grammatical Constructions: Second-Person Singular in Chilean Spanish
Authors: Callaghan Matthew , Travis Catherine E.
Published: December 2020

Structural priming has been described as a measure of association between constructions. Here, we apply priming as a diagnostic to assess the status of the Chilean second-person singular (2sg) voseo, which exists in variation with the more standard tuteo. Despite being the majority variant in informal interactions, Chileans are reported to have little metalinguistic awareness of voseo and they avoid the vos pronoun, in some cases using the tú pronoun with voseo verb forms, leading to proposals that tuteo and voseo are conflated into a single mixed form. The patterning for priming, however, indicates otherwise. Analyses of some 2000 2sg familiar tokens from a corpus of conversational Chilean Spanish reveal that a previous tuteo or voseo favors the repetition of that same form, indicating that speakers do treat these forms as distinct. We also observe that invariable forms with historically tuteo morphology are associated with neither voseo nor tuteo, while the invariable voseo discourse marker cachái ‘you know’ retains a weak association with voseo. Furthermore, while tuteo is favored with a tú subject pronoun, this effect does not override the priming effect, evidence that, even with a tú pronoun, voseo and tuteo are distinct constructions in speakers’ representations.
High-Coordinate Mononuclear Ln(III) Complexes: Synthetic Strategies and Magnetic Properties
Authors: Acharya Joydev , Kalita Pankaj , Chandrasekhar Vadapalli
Published: December 2020

Single-molecule magnets involving monometallic 4f complexes have been investigated extensively in last two decades to understand the factors that govern the slow magnetization relaxation behavior in these complexes and to establish a magneto-structural correlation. The prime goal in this direction is to suppress the temperature independent quantum tunneling of magnetization (QTM) effect via fine-tuning the coordination geometry/microenvironment. Among the various coordination geometries that have been pursued, complexes containing high coordination number around Ln(III) are sparse. Herein, we present a summary of the various synthetic strategies that were used for the assembly of 10- and 12-coordinated Ln(III) complexes. The magnetic properties of such complexes are also described.
Synthesis, NMR Characterization, and Antileukemic Activity of N-Nonanoylpiperazinyl-5α-Androstane-3α,17β-Diol A-Ring Derivatives
Authors: Poirier Donald , Raad Imad , Roy Jenny , Maltais René
Published: December 2020

The combination of an androstane-3,17-diol nucleus and a 2β-N-alkylamidopiperazino sidechain is important for the anticancer activity of a new family of steroid derivatives. As the structure-activity relationship studies have so far been limited to the beta orientation of the substituent at position 2 of the steroid nucleus, a series of analogs (compounds 1–4) were synthesized to investigate the impact on biological activity of A-ring substitution. Nuclear magnetic resonance (NMR) analysis, especially using a series of 2D experiments, such as correlation spectroscopy (COSY), homonuclear Overhauser effect spectroscopy (NOESY), heteronuclear single-quantum correlation (HSQC), and heteronuclear multiple-bond correlation (HMBC) provided crucial information that was found essential in confirming the sidechain position and orientation of compounds 1–4. Assessment of their antiproliferative activity on leukemia HL-60 cells confirmed the best efficiency of the 2β-sidechain/3α-OH orientation (compound 1) compared to the other configurations tested (compounds 2–4).
Eigenvalues of Elliptic Functional Differential Systems via a Birkhoff–Kellogg Type Theorem
Authors: Infante Gennaro
Published: December 2020

Motivated by recent interest on Kirchhoff-type equations, in this short note we utilize a classical, yet very powerful, tool of nonlinear functional analysis in order to investigate the existence of positive eigenvalues of systems of elliptic functional differential equations subject to functional boundary conditions. We obtain a localization of the corresponding non-negative eigenfunctions in terms of their norm. Under additional growth conditions, we also prove the existence of an unbounded set of eigenfunctions for these systems. The class of equations that we study is fairly general and our approach covers some systems of nonlocal elliptic differential equations subject to nonlocal boundary conditions. An example is presented to illustrate the theory.
The Financial Valuation Risk in Pepper Production: The Use of Decoupled Net Present Value
Authors: López-Marín Josefa , Gálvez Amparo , del Amor Francisco M. , Brotons Jose M.
Published: December 2020

Greenhouse peppers are one of the most important crops globally. However, as in any production activity, especially agricultural, they are subject to important risk factors such as price fluctuations, pests, or the use of bad quality water. This article aims to evaluate the viability of these types of crops by using discounted cash flows. Risk evaluation has been carried out through the analysis of pepper plantations for 2016 and 2017. The traditional application of this tool has significant limitations, such as the discount rate to be used or the estimation of future cash flows. However, by using discount functions that decrease over time in combination with decoupled net present value, these limitations are expected to improve. The use of decoupled net present value has permitted an increase in the accuracy and quantification of risks, isolating the main risks such as price drops (EUR 3720 ha−1 year−1) and structural risks (EUR 1622 € ha−1 year−1). The use of decreasing discount functions has permitted a more realistic investment estimation. Finally, the sensitivity analysis shows that decoupled net present value (DNPV) is little affected by changes in interest rates in contrast to traditional net present value (NPV).
Genomic Analysis of Antimicrobial Resistance Genotype-to-Phenotype Agreement in Helicobacter pylori
Authors: Domanovich-Asor Tal , Motro Yair , Khalfin Boris , Craddock Hillary A. , Peretz Avi , Moran-Gilad Jacob
Published: December 2020

Antimicrobial resistance (AMR) in Helicobacter pylori is increasing and can result in treatment failure and inappropriate antibiotic usage. This study used whole genome sequencing (WGS) to comprehensively analyze the H. pylori resistome and phylogeny in order to characterize Israeli H. pylori. Israeli H. pylori isolates (n = 48) underwent antimicrobial susceptibility testing (AST) against five antimicrobials and WGS analysis. Literature review identified 111 mutations reported to correlate with phenotypic resistance to these antimicrobials. Analysis was conducted via our in-house bioinformatics pipeline targeting point mutations in the relevant genes (pbp1A, 23S rRNA, gyrA, rdxA, frxA, and rpoB) in order to assess genotype-to-phenotype correlation. Resistance rates of study isolates were as follows: clarithromycin 54%, metronidazole 31%, amoxicillin 10%, rifampicin 4%, and levofloxacin 2%. Genotype-to-phenotype correlation was inconsistent; for every analyzed gene at least one phenotypically susceptible isolate was found to have a mutation previously associated with resistance. This was also observed regarding mutations commonly used in commercial kits to diagnose AMR in H. pylori cases. Furthermore, 11 novel point mutations associated with a resistant phenotype were detected. Analysis of a unique set of H. pylori isolates demonstrates that inferring resistance phenotypes from WGS in H. pylori remains challenging and should be optimized further.
Growth Inhibition by Amino Acids in Saccharomyces cerevisiae
Authors: Ruiz Stephanie J. , van ’t Klooster Joury S. , Bianchi Frans , Poolman Bert
Published: December 2020

Amino acids are essential metabolites but can also be toxic when present at high levels intracellularly. Substrate-induced downregulation of amino acid transporters in Saccharomyces cerevisiae is thought to be a mechanism to avoid this toxicity. It has been shown that unregulated uptake by the general amino acid permease Gap1 causes cells to become sensitive to amino acids. Here, we show that overexpression of eight other amino acid transporters (Agp1, Bap2, Can1, Dip5, Gnp1, Lyp1, Put4, or Tat2) also induces a growth defect when specific single amino acids are present at concentrations of 0.5–5 mM. We can now state that all proteinogenic amino acids, as well as the important metabolite ornithine, are growth inhibitory to S. cerevisiae when transported into the cell at high enough levels. Measurements of initial transport rates and cytosolic pH show that toxicity is due to amino acid accumulation and not to the influx of co-transported protons. The amino acid sensitivity phenotype is a useful tool that reports on the in vivo activity of transporters and has allowed us to identify new transporter-specific substrates.
Diversity of Methanogens in Animals’ Gut
Authors: Guindo Cheick Oumar , Davoust Bernard , Drancourt Michel , Grine Ghiles
Published: December 2020

Methanogens are members of anaerobe microbiota of the digestive tract of mammals, including humans. However, the sources, modes of acquisition, and dynamics of digestive tract methanogens remain poorly investigated. In this study, we aimed to expand the spectrum of animals that could be sources of methanogens for humans by exploring methanogen carriage in animals. We used real-time PCR, PCR-sequencing, and multispacer sequence typing to investigate the presence of methanogens in 407 fecal specimens collected from nine different mammalian species investigated here. While all the negative controls remained negative, we obtained by PCR-sequencing seven different species of methanogens, of which three (Methanobrevibacter smithii, Methanobrevibacter millerae and Methanomassiliicoccus luminyensis) are known to be part of the methanogens present in the human digestive tract. M. smithii was found in 24 cases, including 12/24 (50%) in pigs, 6/24 (25%) in dogs, 4/24 (16.66%) in cats, and 1/24 (4.16%) in both sheep and horses. Genotyping these 24 M. smithii revealed five different genotypes, all known in humans. Our results are fairly representative of the methanogen community present in the digestive tract of certain animals domesticated by humans, and other future studies must be done to try to cultivate methanogens here detected by molecular biology to better understand the dynamics of methanogens in animals and also the likely acquisition of methanogens in humans through direct contact with these animals or through consumption of the meat and/or milk of certain animals, in particular cows.
Vinification without Saccharomyces: Interacting Osmotolerant and “Spoilage” Yeast Communities in Fermenting and Ageing Botrytised High-Sugar Wines (Tokaj Essence)
Authors: Csoma Hajnalka , Kállai Zoltán , Antunovics Zsuzsa , Czentye Kinga , Sipiczki Matthias
Published: December 2020

The conversion of grape juice to wine starts with complex yeast communities consisting of strains that have colonised the harvested grape and/or reside in the winery environment. As the conditions in the fermenting juice gradually become inhibitory for most species, they are rapidly overgrown by the more adaptable Saccharomyces strains, which then complete the fermentation. However, there are environmental factors that even Saccharomyces cannot cope with. We show that when the sugar content is extremely high, osmotolerant yeasts, usually considered as “spoilage yeasts“, ferment the must. The examination of the yeast biota of 22 botrytised Tokaj Essence wines of sugar concentrations ranging from 365 to 752 g∙L−1 identified the osmotolerant Zygosaccharomyces rouxii, Candida (Starmerella) lactis-condensi and Candida zemplinina (Starmerella bacillaris) as the dominating species. Ten additional species, mostly known as osmotolerant spoilage yeasts or biofilm-producing yeasts, were detected as minor components of the populations. The high phenotypical and molecular (karyotype, mtDNA restriction fragment length polymorphism (RFLP) and microsatellite-primed PCR (MSP-PCR)) diversity of the conspecific strains indicated that diverse clones of the species coexisted in the wines. Genetic segregation of certain clones and interactions (antagonism and crossfeeding) of the species also appeared to shape the fermenting yeast biota.
Pseudouridines of tRNA Anticodon Stem-Loop Have Unexpected Role in Mutagenesis in Pseudomonas sp.
Authors: Tagel Mari , Ilves Heili , Leppik Margus , Jürgenstein Karl , Remme Jaanus , Kivisaar Maia
Published: December 2020

Pseudouridines are known to be important for optimal translation. In this study we demonstrate an unexpected link between pseudouridylation of tRNA and mutation frequency in Pseudomonas species. We observed that the lack of pseudouridylation activity of pseudouridine synthases TruA or RluA elevates the mutation frequency in Pseudomonas putida 3 to 5-fold. The absence of TruA but not RluA elevates mutation frequency also in Pseudomonas aeruginosa. Based on the results of genetic studies and analysis of proteome data, the mutagenic effect of the pseudouridylation deficiency cannot be ascribed to the involvement of error-prone DNA polymerases or malfunctioning of DNA repair pathways. In addition, although the deficiency in TruA-dependent pseudouridylation made P. putida cells more sensitive to antimicrobial compounds that may cause oxidative stress and DNA damage, cultivation of bacteria in the presence of reactive oxygen species (ROS)-scavenging compounds did not eliminate the mutator phenotype. Thus, the elevated mutation frequency in the absence of tRNA pseudouridylation could be the result of a more specific response or, alternatively, of a cumulative effect of several small effects disturbing distinct cellular functions, which remain undetected when studied independently. This work suggests that pseudouridines link the translation machinery to mutation frequency.
Glypican-3-Targeted Alpha Particle Therapy for Hepatocellular Carcinoma
Authors: Bell Meghan M. , Gutsche Nicholas T. , King A. Paden , Baidoo Kwamena E. , Kelada Olivia J. , Choyke Peter L. , Escorcia Freddy E.
Published: December 2020

Glypican-3 (GPC3) is expressed in 75% of hepatocellular carcinoma (HCC), but not normal liver, making it a promising HCC therapeutic target. GC33 is a full-length humanized monoclonal IgG1 specific to GPC3 that can localize to HCC in vivo. GC33 alone failed to demonstrate therapeutic efficacy when evaluated in patients with HCC; however, we posit that cytotoxic functionalization of the antibody with therapeutic radionuclides, may be warranted. Alpha particles, which are emitted by radioisotopes such as Actinium-225 (Ac-225) exhibit high linear energy transfer and short pathlength that, when targeted to tumors, can effectively kill cancer and limit bystander cytotoxicity. Macropa, an 18-member heterocyclic crown ether, can stably chelate Ac-225 at room temperature. Here, we synthesized and evaluated the efficacy of [225Ac]Ac–Macropa–GC33 in mice engrafted with the GPC3-expressing human liver cancer cell line HepG2. Following a pilot dose-finding study, mice (n = 10 per group) were treated with (1) PBS, (2) mass-equivalent unmodified GC33, (3) 18.5 kBq [225Ac]Ac–Macropa–IgG1 (isotype control), (4) 9.25 kBq [225Ac]Ac–Macropa–GC33, and (5) 18.5 kBq [225Ac]Ac–Macropa–GC33. While significant toxicity was observed in all groups receiving radioconjugates, the 9.25 kBq [225Ac]Ac–Macropa–GC33 group demonstrated a modest survival advantage compared to PBS (p = 0.0012) and 18.5 kBq [225Ac]Ac–IgG1 (p = 0.0412). Hematological analysis demonstrated a marked, rapid reduction in white blood cells in all radioconjugate-treated groups compared to the PBS and unmodified GC33 control groups. Our studies highlight a significant disadvantage of using directly-labeled biomolecules with long blood circulation times for TAT. Strategies to mitigate such treatment toxicity include dose fractionation, pretargeting, and using smaller targeting ligands.
On the Rational Drug Design for Hypertension through NMR Spectroscopy
Authors: Chontzopoulou Eleni , Tzakos Andreas G. , Mavromoustakos Thomas
Published: December 2020

Antagonists of the AT1receptor (AT1R) are beneficial molecules that can prevent the peptide hormone angiotensin II from binding and activating the specific receptor causing hypertension in pathological states. This review article summarizes the multifaced applications of solid and liquid state high resolution nuclear magnetic resonance (NMR) spectroscopy in antihypertensive commercial drugs that act as AT1R antagonists. The 3D architecture of these compounds is explored through 2D NOESY spectroscopy and their interactions with micelles and lipid bilayers are described using solid state 13CP/MAS, 31P and 2H static solid state NMR spectroscopy. Due to their hydrophobic character, AT1R antagonists do not exert their optimum profile on the AT1R. Therefore, various vehicles are explored so as to effectively deliver these molecules to the site of action and to enhance their pharmaceutical efficacy. Cyclodextrins and polymers comprise successful examples of effective drug delivery vehicles, widely used for the delivery of hydrophobic drugs to the active site of the receptor. High resolution NMR spectroscopy provides valuable information on the physical-chemical forces that govern these drug:vehicle interactions, knowledge required to get a deeper understanding on the stability of the formed complexes and therefore the appropriateness and usefulness of the drug delivery system. In addition, it provides valuable information on the rational design towards the synthesis of more stable and efficient drug formulations.
Topochemical Engineering of Cellulose—Carboxymethyl Cellulose Beads: A Low-Field NMR Relaxometry Study
Authors: De Wever Pieter , de Oliveira-Silva Rodrigo , Marreiros João , Ameloot Rob , Sakellariou Dimitrios , Fardim Pedro
Published: December 2020

The demand for more ecological, highly engineered hydrogel beads is driven by a multitude of applications such as enzyme immobilization, tissue engineering and superabsorbent materials. Despite great interest in hydrogel fabrication and utilization, the interaction of hydrogels with water is not fully understood. In this work, NMR relaxometry experiments were performed to study bead–water interactions, by probing the changes in bead morphology and surface energy resulting from the incorporation of carboxymethyl cellulose (CMC) into a cellulose matrix. The results show that CMC improves the swelling capacity of the beads, from 1.99 to 17.49, for pure cellulose beads and beads prepared with 30% CMC, respectively. Changes in water mobility and interaction energy were evaluated by NMR relaxometry. Our findings indicate a 2-fold effect arising from the CMC incorporation: bead/water interactions were enhanced by the addition of CMC, with minor additions having a greater effect on the surface energy parameter. At the same time, bead swelling was recorded, leading to a reduction in surface-bound water, enhancing water mobility inside the hydrogels. These findings suggest that topochemical engineering by adjusting the carboxymethyl cellulose content allows the tuning of water mobility and porosity in hybrid beads and potentially opens up new areas of application for this biomaterial.
Application of Ultrasound Combined with Acetic Acid and Peracetic Acid: Microbiological and Physicochemical Quality of Strawberries
Authors: Donatti Leão Alvarenga Priscila , Mileib Vasconcelos Christiane , de São José Jackline Freitas Brilhante
Published: December 2020

This work evaluated the application of organic acids (acetic and peracetic acid) and ultrasound as alternative sanitization methods for improving the microbiological and physicochemical qualities of strawberries. A reduction of up to 2.48 log CFU/g aerobic mesophiles and between 0.89 and 1.45 log CFU/g coliforms at 35 °C was found. For molds and yeasts, significant differences occurred with different treatments and storage time (p < 0.05). Ultrasound treatments in combination with peracetic acid and acetic acid allowed a decimal reduction in molds and yeasts (p < 0.05). All evaluated treatments promoted a significant reduction in the Escherichia coli count (p < 0.05). Scanning electron microscopy revealed fragmented E. coli cells due to treatment with acetic acid and ultrasound. Storage time significantly affected pH, total titratable acidity, total soluble solids and the ratio of the total titratable acidity to the total soluble solids (p < 0.05). Anthocyanin content did not change with treatment or time and generally averaged 13.47 mg anthocyanin/100 g of strawberries on fresh matter. Mass loss was not significantly affected by the applied treatments (p > 0.05). The combination of ultrasound and peracetic acid may be an alternative to chlorine-based compounds to ensure microbiological safety without causing significant changes in the physicochemical characteristics of strawberries.
Discovery of Novel Symmetrical 1,4-Dihydropyridines as Inhibitors of Multidrug-Resistant Protein (MRP4) Efflux Pump for Anticancer Therapy
Authors: Döring Henry , Kreutzer David , Ritter Christoph , Hilgeroth Andreas
Published: December 2020

Despite the development of targeted therapies in cancer, the problem of multidrug resistance (MDR) is still unsolved. Most patients with metastatic cancer die from MDR. Transmembrane efflux pumps as the main cause of MDR have been addressed by developed inhibitors, but early inhibitors of the most prominent and longest known efflux pump P-glycoprotein (P-gp) were disappointing. Those inhibitors have been used without knowledge about the expression of P-gp by the treated tumor. Therefore the use of inhibitors of transmembrane efflux pumps in clinical settings is reconsidered as a promising strategy in the case of the respective efflux pump expression. We discovered novel symmetric inhibitors of the symmetric efflux pump MRP4 encoded by the ABCC4 gene. MRP4 is involved in many kinds of cancer with resistance to anticancer drugs. All compounds showed better activities than the best known MRP4 inhibitor MK571 in an MRP4-overexpressing cell line assay, and the activities could be related to the various substitution patterns of aromatic residues within the symmetric molecular framework. One of the best compounds was demonstrated to overcome the MRP4-mediated resistance in the cell line model to restore the anticancer drug sensitivity as a proof of concept.
Bile Acid Conjugates with Anticancer Activity: Most Recent Research
Authors: Navacchia Maria Luisa , Marchesi Elena , Perrone Daniela
Published: December 2020

The advantages of a treatment modality that combines two or more therapeutic agents in cancer therapy encourages the study of hybrid functional compounds for pharmacological applications. In light of this, we reviewed recent works on hybrid molecules based on bile acids. Due to their biological properties, as well as their different chemical/biochemical reactive moieties, bile acids can be considered very interesting starting molecules for conjugation with natural or synthetic bioactive molecules.
Curcumin Analogue L48H37 Suppresses Human Osteosarcoma U2OS and MG-63 Cells’ Migration and Invasion in Culture by Inhibition of uPA via the JAK/STAT Signaling Pathway
Authors: Lu Ko-Hsiu , Wu Heng-Hsiung , Lin Renn-Chia , Lin Ya-Chiu , Lu Peace Wun-Ang , Yang Shun-Fa , Yang Jia-Sin
Published: December 2020

Osteosarcoma, the most prevalent malignant bone tumor in the pediatric age group, is responsible for the great majority of cancer-associated deaths owing to its highly metastatic potential. The anti-metastatic effects of the new curcumin analogue L48H37 in human osteosarcoma are still unknown; hence, we investigated whether L48H37 represses human osteosarcoma cells’ biological behavior of migratory potential and invasive activities and attempted to delve into its underlying mechanisms. L48H37 up to 5 μM inhibited, without cytotoxicity, the motility, migration, and invasion of human osteosarcoma U2OS and MG-63 cells. In U2OS cells, the human protease array revealed an obvious decrease in urokinase plasminogen activator (uPA) expression after L48H37 treatment, and L48H37 actually reduced the level, protein and mRNA expression, and promoter activity of uPA dose-dependently. L48H37 decreased the phosphorylation of STAT3, JAK1, JAK2, and JAK3 in U2OS cells, but did not affect the phosphorylation of ERK, JNK, p38, and Akt. Using colivelin, an activator of STAT3, the L48H37-induced decrease in uPA and migratory potential could be countered as expected. Collectively, L48H37 represses the invasion and migration capabilities of U2OS and MG-63 cells by the suppression of uPA expression and the inhibition of JAK/STAT signaling. These results suggest that L48H37 may be a potential candidate for anti-metastatic treatment of human osteosarcoma.
Efficient Low-Cost Procedure for Microextraction of Estrogen from Environmental Water Using Magnetic Ionic Liquids
Authors: Berton Paula , Siraj Noureen , Das Susmita , de Rooy Sergio , Wuilloud Rodolfo G. , Warner Isiah M.
Published: December 2020

In this study, three magnetic ionic liquids (MILs) were investigated for extraction of four estrogens, i.e., estrone (E1), estradiol (E2), estriol (E3), and ethinylestradiol (EE2), from environmental water. The cation trihexyl(tetradecyl)phosphonium ([P66614]+), selected to confer hydrophobicity to the resulting MIL, was combined with tetrachloroferrate(III), ferricyanide, and dysprosium thiocyanate to yield ([P66614][FeCl4]), ([P66614]3[Fe(CN)6]), and ([P66614]5[Dy(SCN)8]), respectively. After evaluation of various strategies to develop a liquid–liquid microextraction technique based on synthesized MILs, we placed the MILs onto a magnetic stir bar and used them as extracting solvents. After extraction, the MIL-enriched phase was dissolved in methanol and injected into an HPLC–UV for qualitative and quantitative analysis. An experimental design was used to simultaneously evaluate the effect of select variables and optimization of extraction conditions to maximize the recovery of the analytes. Under optimum conditions, limits of detection were in the range of 0.2 (for E3 and E2) and 0.5 μg L−1 (for E1), and calibration curves exhibited linearity in the range of 1–1000 μg L−1 with correlation coefficients higher than 0.998. The percent relative standard deviation (RSD) was below 5.0%. Finally, this method was used to determine concentration of estrogens in real lake and sewage water samples.
Characteristics of Immunoglobulin M Type Antibodies of Different Origins from the Immunologic and Clinical Viewpoints and Their Application in Controlling Antibody-Mediated Allograft Rejection
Authors: Matsuda Yoshiko , Hiramitsu Takahisa , Li Xiao-kang , Watanabe Takeshi
Published: December 2020

Antibody-mediated allograft rejection (AMR) hinders patient prognosis after organ transplantation. Current studies concerning AMR have mainly focused on the diagnostic value of immunoglobulin G (IgG)-type donor-specific antihuman leukocyte antigen antibodies (DSAs), primarily because of their antigen specificity, whereas the clinical significance of immunoglobulin M (IgM)-type DSAs has not been thoroughly investigated in the context of organ transplantation because of their nonspecificity against antigens. Although consensus regarding the clinical significance and role of IgM antibodies is not clear, as discussed in this review, recent findings strongly suggest that they also have a huge potential in novel diagnostic as well as therapeutic application for the prevention of AMR. Most serum IgM antibodies are known to comprise natural antibodies with low affinity toward antigens, and this is derived from B-1 cells (innate B cells). However, some of the serum IgM-type antibodies reportedly also produced by B-2 cells (conventional B cells). The latter are known to have a high affinity for donor-specific antigens. In this review, we initially discuss how IgM-type antibodies of different origins participate in the pathology of various diseases, directly or through cell surface receptors, complement activation, or cytokine production. Then, we discuss the clinical applicability of B-1 and B-2 cell-derived IgM-type antibodies for controlling AMR with reference to the involvement of IgM antibodies in various pathological conditions.
Comparative Analysis of the Behaviour of Marine Litter in Thermochemical Waste Treatment Processes
Authors: Hee Johann , Schlögel Kai , Lechthaler Simone , Plaster Jacqueline , Bitter Kristina , Blank Lars Mathias , Quicker Peter
Published: December 2020

Plastic in the ocean, especially plastic on the ocean surface is not only researched intensively but also photos and reports rise awareness of the challenge in the general public. While research is concerned with the fate of marine litter in the environment, recycling of these materials after collection is rarely addressed, mainly because there is neither considerable data on composition nor a suggested process to do so. This study is the first to analyse and evaluate chemical recycling (pyrolysis, gasification) and energy recovery (incineration) of marine litter. Two heterogenous marine litter samples from Sylt and Norderney, North Sea, Germany, were analysed, consisting of six different material groups. Agricultural mulch foil was used as reference material. The thermochemical treatment processes were reproduced by thermogravimetric analysis. Furthermore, pyrolysis trials on a semi-technical scale were conducted and the residues were analysed by proximate, ultimate and X-ray fluorescence analysis. The results indicate that heterogeneous and weathered material mixtures can be treated by thermochemical processes. Finally, the pyrolysis condensates are discussed as substrate for biotechnological upcycling. In summary, we present a comprehensive approach from the material characterisation of marine litter to the analysis of three different thermochemical treatment processes and the possibility to use the generated pyrolysis condensate for subsequent upcycling. The data collected form the basis for the evaluation and application of possible treatment options for the collected marine litter.
Study on the Extraction Technology of Candida antarctica Lipase B by Foam Separation
Authors: Shao Wenyao , Lin Ying , Lu Yinghua
Published: December 2020

Candida antarctica Lipase B (CALB) has a wide range of applications in many fields. In this study, Pichia pastoris was used to express CALB for fermentation tank culture. Sodium dodecyl sulfate (SDS) was used as a surfactant, and foam separation technology was used to explore the best experimental conditions for the harvest of CALB. The results showed that the optimal technological conditions for the foam separation and recovery of CALB were as follows: liquid volume was 150 mL, separating gas velocity was 600 mL/min, pH value was 7, and surfactant SDS concentration was 0.5 mg/mL. Under these conditions, the enrichment ratio of CALB was 0.95, and recovery rate R was 80.32%, respectively, indicating that the foam separation technology is feasible to extract lipase B.
Effects of Short-Term Concurrent Training Cessation on the Energy Cost of Running and Neuromuscular Performances in Middle-Distance Runners
Authors: Berryman Nicolas , Mujika Iñigo , Bosquet Laurent
Published: December 2020

Evidence supports the implementation of concurrent strength and running training, within the same mesocycle, to improve performances in middle- and long-distance events. However, very little is known about the effects of concurrent training cessation. The purpose of this investigation was to describe the effects of 4 weeks of explosive strength training cessation after an 8-week concurrent training protocol. Eight runners completed this study, which first included either plyometric (n = 4) or dynamic weight training (n = 4) in addition to the usual running regimen. Explosive strength training was thereafter interrupted for 4 weeks, during which running sessions were maintained. Participants were tested at baseline, after concurrent training and after concurrent training cessation. The results suggest that the energy cost of running improvements observed after the intervention (−5.75%; 95% CI = −8.47 to −3.03) were maintained once explosive strength training was interrupted (−6.31%; 95% CI = −10.30 to −2.32). The results also suggest that neuromuscular performances were maintained after 4 weeks of concurrent training cessation, especially when tests were specific to the training intervention. Furthermore, a 3000m time trial revealed a similar pattern, with improvements after the concurrent mesocycle (−2.40%; 95% CI = −4.65 to −0.16) and after concurrent training cessation (−4.43%; 95% CI = −6.83 to −2.03). Overall, only trivial changes were observed for aerobic endurance and V˙O2peak. Together, these results suggest that short-term explosive strength training cessation might be beneficial and could be considered as a taper strategy for middle-distance runners. However, coaches and athletes must interpret these results cautiously considering the study’s low sample size and the very limited available literature in this domain.