bluuebunny/crossref_metadata_2025 huggingface dataset:
LC-ESI-QTOF-MS/MS Characterization of Seaweed Phenolics and Their Antioxidant Potential
Authors: Zhong Biming , Robinson Nicholas A. , Warner Robyn D. , Barrow Colin J. , Dunshea Frank R. , Suleria Hafiz A.R.
Published: June 2020

Seaweed is an important food widely consumed in Asian countries. Seaweed has a diverse array of bioactive compounds, including dietary fiber, carbohydrate, protein, fatty acid, minerals and polyphenols, which contribute to the health benefits and commercial value of seaweed. Nevertheless, detailed information on polyphenol content in seaweeds is still limited. Therefore, the present work aimed to investigate the phenolic compounds present in eight seaweeds [Chlorophyta (green), Ulva sp., Caulerpa sp. and Codium sp.; Rhodophyta (red), Dasya sp., Grateloupia sp. and Centroceras sp.; Ochrophyta (brown), Ecklonia sp., Sargassum sp.], using liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS/MS). The total phenolic content (TPC), total flavonoid content (TFC) and total tannin content (TTC) were determined. The antioxidant potential of seaweed was assessed using a 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay, a 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) free radical scavenging assay and a ferric reducing antioxidant power (FRAP) assay. Brown seaweed species showed the highest total polyphenol content, which correlated with the highest antioxidant potential. The LC-ESI-QTOF-MS/MS tentatively identified a total of 54 phenolic compounds present in the eight seaweeds. The largest number of phenolic compounds were present in Centroceras sp. followed by Ecklonia sp. and Caulerpa sp. Using high-performance liquid chromatography-photodiode array (HPLC-PDA) quantification, the most abundant phenolic compound was p-hydroxybenzoic acid, present in Ulva sp. at 846.083 ± 0.02 μg/g fresh weight. The results obtained indicate the importance of seaweed as a promising source of polyphenols with antioxidant properties, consistent with the health potential of seaweed in food, pharmaceutical and nutraceutical applications.
Molecular Dynamics-Based Cohesive Zone Model for Mg/Mg17Al12 Interface
Authors: Zhuo Xiao Ru , Ma Aibin
Published: June 2020

The fracture of the Mg/Mg17Al12 interface was investigated by molecular dynamics simulations. The interface crack extends in a brittle manner without noticeable plasticity. The distributions of normal stress and separation along the interface were examined to render a quantitative picture of the fracture process. A normal traction–separation curve was extracted from simulation and compared with three cohesive zone models, i.e., cubic polynomial cohesive zone model, exponential cohesive zone model, and bilinear cohesive zone model. The exponential cohesive zone model exhibits the best agreement with simulation results, followed by the bilinear cohesive zone model.
Quality Assessment Method Based on a Spectrometer in Laser Beam Welding Process
Authors: Yu Jiyoung , Lee Huijun , Kim Dong-Yoon , Kang Munjin , Hwang Insung
Published: June 2020

For the automation of a laser beam welding (LBW) process, the weld quality must be monitored without destructive testing, and the quality must be assessed. A deep neural network (DNN)-based quality assessment method in spectrometry-based LBW is presented in this study. A spectrometer with a response range of 225–975 nm is designed and fabricated to measure and analyze the light reflected from the welding area in the LBW process. The weld quality is classified through welding experiments, and the spectral data are thus analyzed using the spectrometer, according to the welding conditions and weld quality classes. The measured data are converted to RGB (red, green, blue) values to obtain standardized and simplified spectral data. The weld quality prediction model is designed based on DNN, and the DNN model is trained using the experimental data. It is seen that the developed model has a weld-quality prediction accuracy of approximately 90%.
Functional Associations and Resilience in Microbial Communities
Authors: Avila-Jimenez Maria-Luisa , Burns Gavin , He Zhili , Zhou Jizhong , Hodson Andrew , Avila-Jimenez Jose-Luis , Pearce David
Published: June 2020

Microbial communities have inherently high levels of metabolic flexibility and functional redundancy, yet the structure of microbial communities can change rapidly with environmental perturbation. To understand whether such changes observed at the taxonomic level translate into differences at the functional level, we analyzed the structure of taxonomic and functional gene distribution across Arctic and Antarctic locations. Taxonomic diversity (in terms of alpha diversity and species richness) differed significantly with location. However, we found that functional genes distributed evenly across bacterial networks and that this functional distribution was also even across different geographic locations. For example, on average 15% of the functional genes were related to carbon cycling across all bacterial networks, slightly over 21% of the genes were stress-related and only 0.5% of the genes were linked to carbon degradation functions. In such a distribution, each bacterial network includes all of the functional groups distributed following the same proportions. However, the total number of functional genes that is included in each bacterial network differs, with some clusters including many more genes than others. We found that the proportion of times a specific gene must occur to be linked to a specific cluster is 8%, meaning the relationship between the total number of genes in the cluster and the number of genes per function follows a linear pattern: smaller clusters require a gene to appear less frequently to get fixed within the cluster, while larger clusters require higher gene frequencies. We suggest that this mechanism of functional association between equally rare or equally abundant genes could have implications for ecological resilience, as non-dominant genes also associate in fully functioning ecological networks, potentially suggesting that there are always pre-existing functional networks available to exploit new ecological niches (where they can become dominant) as they emerge; for example, in the case of rapid or sudden environmental change. Furthermore, this pattern did not correlate with taxonomic distribution, suggesting that bacteria associate based on functionality and this is independent of its taxonomic position. Our analyses based on ecological networks also showed no clear evidence of recent environmental impact on polar marine microbial communities at the functional level, unless all communities analyzed have changed exactly in the same direction and intensity, which is unlikely given we are comparing areas changing at different rates.
Lactic Acid Bacteria as Antibacterial Agents to Extend the Shelf Life of Fresh and Minimally Processed Fruits and Vegetables: Quality and Safety Aspects
Authors: Agriopoulou Sofia , Stamatelopoulou Eygenia , Sachadyn-Król Monika , Varzakas Theodoros
Published: June 2020

Eating fresh fruits and vegetables is, undoubtedly, a healthy habit that should be adopted by everyone (particularly due to the nutrients and functional properties of fruits and vegetables). However, at the same time, due to their production in the external environment, there is an increased risk of their being infected with various pathogenic microorganisms, some of which cause serious foodborne illnesses. In order to preserve and distribute safe, raw, and minimally processed fruits and vegetables, many strategies have been proposed, including bioprotection. The use of lactic acid bacteria in raw and minimally processed fruits and vegetables helps to better maintain their quality by extending their shelf life, causing a significant reduction and inhibition of the action of important foodborne pathogens. The antibacterial effect of lactic acid bacteria is attributed to its ability to produce antimicrobial compounds, including bacteriocins, with strong competitive action against many microorganisms. The use of bacteriocins, both separately and in combination with edible coatings, is considered a very promising approach for microbiological quality, and safety for postharvest storage of raw and minimally processed fruits and vegetables. Therefore, the purpose of the review is to discuss the biopreservation of fresh fruits and vegetables through the use of lactic acid bacteria as a green and safe technique.
Effects of Crude β-Glucosidases from Issatchenkia terricola, Pichia kudriavzevii, Metschnikowia pulcherrima on the Flavor Complexity and Characteristics of Wines
Authors: Zhang Wenxia , Zhuo Xuanhan , Hu Lanlan , Zhang Xiuyan
Published: June 2020

To investigate the effects of crude β-glucosidases from Issatchenkia terricola SLY-4 (SLY-4E), Pichia kudriavzevii F2-24 (F2-24E), and Metschnikowia pulcherrima HX-13 (HX-13E) on flavor complexity and characteristics of wines, grape juice was fermented by Saccharomyces cerevisiae with the addition of SLY-4E, F2-24E and HX-13E, respectively. The growth and sugar consumption kinetics of S. cerevisiae, the physicochemical characteristics, the volatile compounds, and the sensory dimensions of wines were analyzed. Results showed that adding SLY-4E, F2-24E, and HX-13E into must had no negative effect on the fermentation and physicochemical characteristics of wines, but increased the content of terpenes, esters, and fatty acids, while decreased the C6 compound content. Each wine had its typical volatile compound profiles. Adding SLY-4E or F2-24E into must could significantly improve the flavor complexity and characteristics of wines. These results would provide not only an approach to improve flavor complexity and characteristics of wines, but also references for application of β-glucosidases from other sources.
Total Dietary Antioxidant Intake Including Polyphenol Content: Is It Capable to Fight against Increased Oxidants within the Body of Ultra-Endurance Athletes?
Authors: Devrim-Lanpir Aslı , Bilgic Pelin , Kocahan Tuğba , Deliceoğlu Gökhan , Rosemann Thomas , Knechtle Beat
Published: June 2020

The role of dietary antioxidants on exhaustive exercise-induced oxidative stress has been well investigated. However, the contribution of total dietary antioxidant capacity on exogenous antioxidant defense and exercise performance has commonly been disregarded. The aims of the present investigation were to examine (i) the effects of dietary total antioxidant intake on body antioxidant mechanisms, and (ii) an exhaustive exercise-induced oxidative damage in ultra-endurance athletes. The study included 24 ultra-marathon runners and long-distance triathletes (12 male and 12 female) who underwent an acute exhaustive exercise test (a cycle ergometer (45 min at 65% VO2max) immediately followed by a treadmill test (75% VO2max to exhaustion). Oxidative stress-related biomarkers (8-isoprostaglandin F2alpha (8-iso PGF2a), total oxidant status (TOS, total antioxidant status (TAS)) in plasma were collected before and after exercise. Oxidative stress index was calculated to assess the aspect of redox balance. Blood lactate concentrations and heart rate were measured at the 3rd and 6th min after exercise. Dietary antioxidant intake was calculated using the ferric reducing ability of plasma (FRAP) assay. Dietary total antioxidant intake of the subjects was negatively correlated with pre-exercise TOS concentrations (rs = −0.641 in male, and rs = −0.741 in females) and post- vs. pre- (∆) 8-iso PGF2a levels (rs = −0.702 in male; p = 0.016, and rs = −0.682 in females; p = 0.024), and positively correlated with ∆ TAS concentrations (rs = 0.893 in males; p = 0.001, and rs = 0.769 in females; p = 0.002) and post- exercise lactate concentrations (rs = 0.795 for males; p = 0.006, and rs = 0.642 for females; p = 0.024). A positive meaningful (p = 0.013) interaction was observed between time at exhaustion and dietary antioxidant intake (rs = 0.692) in males, but not in females. In conclusion, the determination of total dietary antioxidant intake in ultra-endurance athletes may be crucial for gaining a better perspective on body antioxidant defense against exhaustive exercise-induced oxidative stress. However, the effects of dietary antioxidant on exercise performance and recovery rate needs further investigation.
Results of a Mass Media Campaign in South Africa to Promote a Sugary Drinks Tax
Authors: Murukutla Nandita , Cotter Trish , Wang Shuo , Cullinan Kerry , Gaston Fathima , Kotov Alexey , Maharjan Meena , Mullin Sandra
Published: June 2020

Background: In South Africa, the increased consumption of sugary drinks has been associated with increased obesity rates. Mass media campaigns can play a crucial role in improving knowledge, shifting attitudes, and building support for government action on reducing sugary drink consumption. No study to date has evaluated the effectiveness of mass media campaigns on the health harms of sugary drinks in South Africa. Objective: The purpose of this study was to evaluate the impact of a mass media campaign on knowledge and attitudes around sugary drinks and on public support for a proposed tax on sugary drinks in South Africa. Methods: The “Are You Drinking Yourself Sick?” campaign aired in South Africa from October 2016 to June 2017 to shift attitudes toward sugary drinks, build personal risk perceptions of the health harms of consuming sugary drinks, and build public support for a proposed tax on sugary drinks. Campaign impact was measured in representative cross-sectional household surveys of adults ages 18 to 56. The surveys were conducted just prior to the launch of the campaign (N = 1000), from October 7 to 10, 2016, and immediately following its conclusion (N = 1000), from July 12 to 21, 2017. Campaign impact was assessed by comparing changes from the pre-campaign to the post-campaign period on key outcome indicators. In addition, the effect of campaign awareness was analyzed in logistic regression analysis of the post-campaign data. Results: The campaign was recalled by 55% of survey respondents, and 78% of campaign-aware respondents said that the campaign’s main message was “drinking sugary drinks can make you sick.” There were significant changes from the pre- to the post-campaign period in knowledge that sugary drink consumption can lead to obesity and related health problems and that sugary drinks contribute toward the obesity problem in South Africa. Campaign awareness was also significantly associated with increases in knowledge about the harms of sugary drinks, and in particular, on government action, including the proposed tax on sugary drinks. Discussion: Media campaigns are an effective intervention for obesity prevention. In addition to improving knowledge and shifting attitudes, media campaigns can effectively build public support for strong government action and therefore must be a component of a comprehensive obesity prevention approach.
The Potential Effects of Human Milk on Morbidity in Very-Low-Birth-Weight Preterm Infants
Authors: Roggero Paola , Liotto Nadia , Amato Orsola , Mosca Fabio
Published: June 2020

Improvements in quality of care have led to a significant reduction in mortality and morbidity in preterm infants, especially very-low-birth-weight (VLBW) infants [...]
Aspartame and Phe-Containing Degradation Products in Soft Drinks across Europe
Authors: van Vliet Kimber , Melis Elise S. , de Blaauw Pim , van Dam Esther , Maatman Ronald G. H. J. , Abeln David , van Spronsen Francjan J. , Heiner-Fokkema M. Rebecca
Published: June 2020

Phenylketonuria and tyrosinemia type 1 are treated with dietary phenylalanine (Phe) restriction. Aspartame is a Phe-containing synthetic sweetener used in many products, including many ‘regular’ soft drinks. Its amount is (often) not declared; therefore, patients are advised not to consume aspartame-containing foods. This study aimed to determine the variation in aspartame concentrations and its Phe-containing degradation products in aspartame-containing soft drinks. For this, an LC–MS/MS method was developed for the analysis of aspartame, Phe, aspartylphenylalanine, and diketopiperazine in soft drinks. In total, 111 regularly used soft drinks from 10 European countries were analyzed. The method proved linear and had an inter-assay precision (CV%) below 5% for aspartame and higher CVs% of 4.4–49.6% for the degradation products, as many concentrations were at the limit of quantification. Aspartame and total Phe concentrations in the aspartame-containing soft drinks varied from 103 to 1790 µmol/L (30–527 mg/L) and from 119 to 2013 µmol/L (20–332 mg/L), respectively, and were highly variable among similar soft drinks bought in different countries. Since Phe concentrations between drinks and countries highly vary, we strongly advocate the declaration of the amount of aspartame on soft drink labels, as some drinks may be suitable for consumption by patients with Phe-restricted diets.
Sequence Analysis of New Tuf Molecular Types of ‘Candidatus Phytoplasma Solani’ in Iranian Vineyards
Authors: Jamshidi Elham , Murolo Sergio , Salehi Mohammad , Romanazzi Gianfranco
Published: June 2020

Grapevine Bois noir (BN) is caused by ‘Candidatus Phytoplasma solani’ (‘Ca. P. solani’) and is one of the most important phytoplasma diseases in the Euro-Mediterranean viticultural areas. The epidemiology of BN can include grapevine as a plant host and is usually transmitted via sap-sucking insects that inhabit herbaceous host plants. Tracking the spread of ‘Ca. P. solani’ strains is of great help for the identification of plant reservoirs and insect vectors involved in local BN outbreaks. The molecular epidemiology of ‘Ca. P. solani’ is primarily based on sequence analysis of the tuf housekeeping gene (which encodes elongation factor Tu). In this study, molecular typing of tuf, through restriction fragment length polymorphism and sequencing, was carried out on grapevine samples from Iranian vineyards. According to the molecular characterization, three molecular types—tuf b1, tuf b5 and tuf b6—were found, with tuf b1 being the most prominent. These data provide further knowledge of tuf gene diversity and question the ecological role of such “minor” tuf types in Iranian vineyards, which have been detected only in grapevines.
Asymptomatic Strongyloidiasis among Latin American Migrants in Spain: A Community-Based Approach
Authors: Ramos-Sesma Violeta , Navarro Miriam , Llenas-García Jara , Gil-Anguita Concepción , Torrús-Tendero Diego , Wikman-Jorgensen Philip , Amador-Prous Concepción , Ventero-Martín María-Paz , Garijo-Sainz Ana-María , García-López María , Pujades-Tárraga Ana-Isabel , Bernal-Alcaraz Cristina , Santonja Antonio , Guevara-Hernández Pedro , Flores-Chávez María , Saugar José-María , Ramos-Rincón José-Manuel ,   Corazones Sin Chagas Platform
Published: June 2020

Strongyloides stercoralis infection is frequently underdiagnosed since many infections remain asymptomatic. Aim: To estimate the prevalence and characteristics of asymptomatic S. stercoralis infection in Latin American migrants attending a community-based screening program for Chagas disease in Spain. Methodology: Three community-based Chagas disease screening campaigns were performed in Alicante (Spain) in 2016, 2017, and 2018. Serological testing for S. stercoralis infection was performed using a non-automatized IVD-ELISA detecting IgG (DRG Instruments GmbH, Marburg, Germany). Results: Of the 616 migrants from Central and South America who were screened, 601 were included in the study: 100 children and adolescents (<18 years of age) and 501 adults. Among the younger group, 6 participants tested positive (prevalence 6%, 95% confidence interval [CI] 2.5% to 13.1%), while 60 adults did so (prevalence 12%, 95% CI 9.3% to 15.3%). S. stercoralis infection was more common in men than in women (odds ratio adjusted [ORa] 2.28, 95% CI 1.289 to 4.03) and in those from Bolivia (ORa 2.03, 95% CI 1.15 to 3.59). Prevalence increased with age (ORa 1.02, 95% CI 0.99 to 1.05). In contrast, a university education had a protective effect (ORa 0.29, 95% CI 0.31 to 0.88). Forty-one (41/66; 62.1%) of the total cases of S. stercoralis infection were treated at the health care center. Positive stool samples were observed in 19.5% of the followed-up positive cases. Conclusion: Incorporating serological screening for S. stercoralis into community-based screening for Chagas disease is a useful intervention to detect asymptomatic S. stercoralis infection in Central and South American migrants and an opportunity to tackle neglected tropical diseases in a transversal way. The remaining challenge is to achieve patients’ adherence to the medical follow-up.
Controlled Delivery of Insulin-like Growth Factor-1 from Bioactive Glass-Incorporated Alginate-Poloxamer/Silk Fibroin Hydrogels
Authors: Min Qing , Yu Xiaofeng , Liu Jiaoyan , Zhang Yuchen , Wan Ying , Wu Jiliang
Published: June 2020

Thermosensitive alginate–poloxamer (ALG–POL) copolymer with an optimal POL content was synthesized, and it was used to combine with silk fibroin (SF) for building ALG–POL/SF hydrogels with dual network structure. Mesoporous bioactive glass (BG) nanoparticles (NPs) with a high level of mesoporosity and large pore size were prepared and they were employed as a vehicle for loading insulin-like growth factor-1 (IGF-1). IGF-1-loaded BG NPs were embedded into ALG–POL/SF hydrogels to achieve the controlled delivery of IGF-1. The resulting IGF-1-loaded BG/ALG–POL/SF gels were found to be injectable with their sol-gel transition near physiological temperature and pH. Rheological measurements showed that BG/ALG–POL/SF gels had their elastic modulus higher than 5kPa with large ratio of elastic modulus to viscous modulus, indicative of their mechanically strong features. The dry BG/ALG–POL/SF gels were seen to be highly porous with well-interconnected pore characteristics. The gels loaded with varied amounts of IGF-1 showed abilities to administer IGF-1 release in approximately linear manners for a few weeks while effectively preserving the bioactivity of encapsulated IGF-1. Results suggest that such constructed BG/ALG–POL/SF gels can function as a promising injectable biomaterial for bone tissue engineering applications.
RGD-PEG-PLA Delivers MiR-133 to Infarct Lesions of Acute Myocardial Infarction Model Rats for Cardiac Protection
Authors: Sun Bixi , Liu Shuwen , Hao Rubin , Dong Xinyue , Fu Lanbo , Han Bing
Published: June 2020

Studies have shown that microRNA-133 (miR-133) plays a positive role in the growth of cardiac myocytes, the maintenance of cardiac homeostasis, and the recovery of cardiac function, which is of great significance for the recovery of acute myocardial infarction. However, the delivery of miRNA to the site of action remains a challenge at present. The purpose of this study was to design an ideal carrier to facilitate the delivery of miR-133 to the infarct lesion for cardiac protection. A disease model was constructed by ligating the left anterior descending coronary artery of rats, and polyethylene glycol (PEG)-polylactic acid (PLA) nanoparticles modified with arginine-glycine-aspartic acid tripeptide (RGD) carrying miR-133 were injected via the tail vein. The effects of miR-133 were evaluated from multiple perspectives, including cardiac function, blood indexes, histopathology, and myocardial cell apoptosis. The results showed that RGD-PEG-PLA maintained a high level of distribution in the hearts of model rats, indicating the role of the carrier in targeting the heart infarction lesions. RGD-PEG-PLA/miR-133 alleviated cardiac histopathological changes, reduced the apoptosis of cardiomyocytes, and reduced the levels of factors associated with myocardial injury. Studies on the mechanism of miR-133 by immunohistochemistry and polymerase chain reaction demonstrated that the expression level of Sirtuin3 (SIRT3) was increased and that the expression of adenosine monophosphate activated protein kinase (AMPK) decreased in myocardial tissue. In summary, the delivery of miR-133 by RGD-PEG-PLA carrier can achieve cardiac lesion accumulation, thereby improving the cardiac function damage and reducing the myocardial infarction area. The inhibition of cardiomyocyte apoptosis, inflammation, and oxidative stress plays a protective role in the heart. The mechanism may be related to the regulation of the SIRT3/AMPK pathway.
Mesenchymal Stem Cell Derived Biocompatible Membrane Vesicles Demonstrate Immunomodulatory Activity Inhibiting Activation and Proliferation of Human Mononuclear Cells
Authors: Gomzikova Marina , Kletukhina Sevindzh , Kurbangaleeva Sirina , Neustroeva Olga , Vasileva Olga , Garanina Ekaterina , Khaiboullina Svetlana , Rizvanov Albert
Published: June 2020

Immune-mediated diseases are characterized by abnormal activity of the immune system. The cytochalasin B-induced membrane vesicles (CIMVs) are innovative therapeutic instruments. However, the immunomodulating activity of human mesenchymal stem cell (MSC)-derived CIMVs (CIMVs-MSCs) remains unknown. Therefore, we sought to investigate the immunological properties of CIMVs-MSCs and evaluate their effect on human peripheral blood mononuclear cells (PBMCs). We found that CIMVs-MSCs are primarily uptaken by monocytes and B-cells. Additionally, we demonstrated that CIMVs-MSCs inhibit phytohemagglutinin (PHA)-induced proliferation of PBMCs, with more pronounced effect on T-lymphocytes expansion as compared to that of B-cells. In addition, activation of T-helpers (CD4+CD25+), B-cells (CD19+CD25+), and T-cytotoxic lymphocytes (CD8+CD25+) was also significantly suppressed by CIMVs-MSCs. Additionally, CIMVs-MSCs decreased secretion of epidermal growth factor (EGF) and pro-inflammatory Fractalkine in a population of PBMCs, while the releases of FGF-2, G-CSF, anti-inflammatory GM-CSF, MCP-3, anti-inflammatory MDC, anti-inflammatory IL-12p70, pro-inflammatory IL-1b, and MCP-1 were increased. We analyzed the effect of CIMVs-MSCs on an isolated population of CD4+ and CD8+ T-lymphocytes and demonstrated their different immune response and cytokine secretion. Finally, we observed that no xenogeneic nor allogeneic transplantation of CIMVs induced an immune response in mice. Our data suggest that CIMVs-MSCs have immunosuppressive properties, are potential agents for immunomodulating treatment, and are worthy of further investigation.
Identification of Cell-Surface Proteins Endocytosed by Human Brain Microvascular Endothelial Cells In Vitro
Authors: Ito Shingo , Oishi Mariko , Ogata Seiryo , Uemura Tatsuki , Couraud Pierre-Olivier , Masuda Takeshi , Ohtsuki Sumio
Published: June 2020

Cell-surface proteins that can endocytose into brain microvascular endothelial cells serve as promising candidates for receptor-mediated transcytosis across the blood–brain barrier (BBB). Here, we comprehensively screened endocytic cell-surface proteins in hCMEC/D3 cells, a model of human brain microvascular endothelial cells, using surface biotinylation methodology and sequential window acquisition of all theoretical fragment-ion spectra-mass spectrometry (SWATH-MS)-based quantitative proteomics. Using this method, we identified 125 endocytic cell-surface proteins from hCMEC/D3 cells. Of these, 34 cell-surface proteins were selectively internalized into human brain microvascular endothelial cells, but not into human umbilical vein endothelial cells (HUVECs), a model of human peripheral microvascular endothelial cells. Two cell-surface proteins, intercellular adhesion molecule-1 (ICAM1) and podocalyxin (PODXL), were identified as BBB-localized endocytic cell-surface proteins in humans, using open mRNA and protein databases. Immunohistochemical evaluation confirmed PODXL expression in the plasma membrane of hCMEC/D3 cells and revealed that anti-PODXL antibody-labeled cell-surface PODXL internalized into hCMEC/D3 cells. Immunohistochemistry further revealed that PODXL is localized at the luminal side of human brain microvessels, supporting its potential suitability for translational applications. In conclusion, our findings highlight novel endocytic cell-surface proteins capable of internalizing into human brain microvascular endothelial cells. ICAM1 or PODXL targeted antibody or ligand-labeled biopharmaceuticals and nanocarriers may provide effective targeted delivery to the brain across the BBB for the treatment of central nervous system (CNS) diseases.
Rapidly Separable Micropillar Integrated Dissolving Microneedles
Authors: Jung Chung-ryong , Lahiji Shayan Fakhraei , Kim Youseong , Kim Hyeonjun , Jung Hyungil
Published: June 2020

Dissolving microneedle (DMN) patches were developed as efficient and patient-friendly transdermal delivery systems for biopharmaceuticals. However, recent studies have confirmed that the efficiency of DMNs to deliver biopharmaceuticals is highly reduced because of incomplete insertion caused by the stiffness and elastic properties of the skin. Therefore, micropillar integrated DMNs were developed to overcome the insertion limitations of DMN patches. Although micropillars were designed as integrated applicators to implant DMNs across the skin, they can also become inserted into the skin, leading to skin injury and inflammation. Herein, we have developed a separable micropillar integrated DMN (SPDMN) capable of inserting DMNs across the skin with high efficiency while minimizing skin injury risk through the introduction of a safety ring feature. Unlike previously developed systems, the SPDMN does not require continuous skin attachment and can be detached immediately post-application, leaving DMNs implanted inside the skin. Altogether, the findings of this study lead to the development of a quick, safe, and efficient DMN-based drug delivery platform.
Manipulation of Medicinal Products for Oral Administration to Paediatric Patients at a German University Hospital: An Observational Study
Authors: Zahn Julia , Hoerning André , Trollmann Regina , Rascher Wolfgang , Neubert Antje
Published: June 2020

Pharmacotherapy in children requires medicinal products in age-appropriate dosage forms and flexible dose strengths. Healthcare professionals often encounter a lack of licensed and commercially available formulations, which results in the need for manipulation. This study aimed to investigate the nature, frequency and preventability of the manipulation of medicinal products before oral drug administration to paediatric inpatients in Germany. A prospective, direct observational approach was used. Two thousand and three medication preparation processes (MPP) in 193 patients were included in the analysis. Medicines were manipulated in 37% of oral administrations, affecting 57% of the patients. The percentage of manipulations was highest in infants/toddlers (42%) and lowest in adolescents (31%). Antiepileptics were most frequently manipulated (27%), followed by vitamins (20%) and drugs for acid-related disorders (13%). Fifty-six per cent of all manipulations were off-label. In 71% of these, no alternative appropriate medicinal product was commercially available. These results demonstrate that the manipulation of medicinal products before oral administration is common in paediatric wards in Germany. About half of the manipulations were off-label, indicating that no suitable formulation was available. Evidence-based guidelines for manipulations are required, with the overall aim of improving the safety of paediatric drug therapy.
Novel Cleaning-in-Place Strategies for Pharmaceutical Hot Melt Extrusion
Authors: Spoerk Martin , Koutsamanis Ioannis , Matić Josip , Eder Simone , Patricia Alva Zúñiga Carolina , Poms Johannes , Urich Jesús Alberto Afonso , Andreína Lara García Raymar , Nickisch Klaus , Eggenreich Karin , Berghaus Andreas , Reusch Kathrin , Relle Yorick , Khinast Johannes , Paudel Amrit
Published: June 2020

To avoid any type of cross-contamination, residue-free production equipment is of utmost importance in the pharmaceutical industry. The equipment cleaning for continuous processes such as hot melt extrusion (HME), which has recently gained popularity in pharmaceutical applications, necessitates extensive manual labour and costs. The present work tackles the HME cleaning issue by investigating two cleaning strategies following the extrusion of polymeric formulations of a hormonal drug and for a sustained release formulation of a poorly soluble drug. First, an in-line quantification by means of UV–Vis spectroscopy was successfully implemented to assess very low active pharmaceutical ingredient (API) concentrations in the extrudates during a cleaning procedure for the first time. Secondly, a novel in-situ solvent-based cleaning approach was developed and its usability was evaluated and compared to a polymer-based cleaning sequence. Comparing the in-line data to typical swab and rinse tests of the process equipment indicated that inaccessible parts of the equipment were still contaminated after the polymer-based cleaning procedure, although no API was detected in the extrudate. Nevertheless, the novel solvent-based cleaning approach proved to be suitable for removing API residue from the majority of problematic equipment parts and can potentially enable a full API cleaning-in-place of a pharmaceutical extruder for the first time.
The Mbeya Antimicrobial Stewardship Team: Implementing Antimicrobial Stewardship at a Zonal-Level Hospital in Southern Tanzania
Authors: Hall Jeffrey W. , Bouchard Jeannette , Bookstaver P. Brandon , Haldeman Matthew S. , Kishimbo Peter , Mbwanji Godlove , Mwakyula Issakwisa , Mwasomola Davance , Seddon Megan , Shaffer Mark , Shealy Stephanie C. , Nsojo Anthony
Published: June 2020

Background: In 2017, Mbeya Zonal Referral Hospital (MZRH) and the University of South Carolina (UofSC) agreed to collaboratively strengthen antimicrobial prescribing in the southern highlands of Tanzania and train a new generation of clinicians in responsible antimicrobial use. Methods: Key stakeholders and participants were identified and the Mbeya Antimicrobial Stewardship Team (MAST) was created. The team identified assets brought by the collaborators, and four investigations of baseline needs were developed. These investigations included (a) a baseline clinician survey regarding antimicrobial resistance and stewardship, (b) a serial chart review of inpatient antimicrobial prescribing practices, (c) an investigation of antimicrobial resistance rates using existing isolates at the MZRH laboratory, and (d) a survey of antimicrobial availability at community pharmacies in the city. Results: 91% of physicians believe antimicrobial resistance is problem in Tanzania, although only 29% of physicians were familiar with the term “antimicrobial stewardship”. Escherichia coli isolates had resistance rates of over 60% to the commonly used agents ciprofloxacin, trimethoprim-sulfamethoxazole, and ceftriaxone. Thirteen out of 14 community pharmacies offered over-the-counter antibiotics for upper respiratory symptoms. Conclusions: International antimicrobial stewardship collaborations can successfully identify opportunities and needs. Evaluating the team’s efforts to improve patient outcomes will be essential.
What Is Physical Information?
Authors: Krzanowski Roman
Published: June 2020

This paper presents the concept of physical information, and it discusses what physical information is, and how it can be defined. The existence of physical information has been discussed in several studies which recognize that properties of information are characteristic of physical phenomena. That is, information has an objective existence, a lack of meaning, and can be quantified. In addition, these studies recognize how a phenomenon that is denoted as physical information can be expressed as an organization of natural or artificial entities. This paper argues that concepts of (abstract) information that are associated with meaning also depend (to a substantial degree) on physical information.
Nitric Oxide Pre-Treatment Advances Seed Germination and Alleviates Copper-Induced Photosynthetic Inhibition in Indian Mustard
Authors: Rather Bilal A. , Mir Iqbal R. , Masood Asim , Anjum Naser A. , Khan Nafees A.
Published: June 2020

This investigation tested the efficiency of nitric oxide (NO) in alleviation of Cu-induced adverse impacts on seed germination and photosynthesis in Indian mustard (Brassica juncea L.). Pre-treatment of B. juncea seeds with sodium nitroprusside (SNP; NO donor) significantly improved the seed germination rate and also alleviated Cu-accrued oxidative stress. However, in the absence of NO, Cu caused a higher reduction in seed germination rate. The presence of NO strengthened the antioxidant defense system (glutathione reductase, ascorbate peroxidase, and superoxide dismutase) and thereby sustained the lower lipid peroxidation, reduced H2O2 content, and thiobarbituric acid reactive substances in Cu-exposed seeds. NO pre-treated seeds also retained a higher amylase activity and exhibited an improved seed germination rate. This effect of NO under Cu stress was also seen in plants originated from the NO pre-treated seeds, where the role of NO pre-treatment was reflected in the improved photosynthetic potential of B. juncea. Overall, NO pre-treatment not only improved the germination rate in seeds but also carried its effects in the grown seedlings evidenced as improved photosynthesis and growth. Potential mechanisms involved in the action of NO pre-treatment included NO-mediated significant strengthening of the antioxidant defense system and decreases in Cu-caused oxidative stress parameters.
Easy-to-Use InDel Markers for Genetic Mapping between Col-0 and Ler-0 Accessions of Arabidopsis thaliana
Authors: Tanaka Takahiro , Nishii Yuichi , Matsuo Hirotoshi , Takahashi Taku
Published: June 2020

Map-based gene cloning has played a key role in many genetic studies using the model plant, Arabidopsis thaliana. In the post- next generation sequencing era, identification of point mutations and their corresponding genes is increasingly becoming a powerful and important approach to define plant gene function. To perform initial mapping experiments efficiently on Arabidopsis mutants, enrichment of easy-to-use and reliable polymorphic DNA markers would be desirable. We present here a list of InDel polymorphic markers between Col-0 and Ler-0 accessions that can be detected in standard agarose gel electrophoresis.
Transcriptome Analysis of Wounding in the Model Grass Lolium temulentum
Authors: Dombrowski James E. , Kronmiller Brent A. , Hollenbeck Vicky , Martin Ruth C.
Published: June 2020

For forage and turf grasses, wounding is a predominant stress that often results in extensive loss of vegetative tissues followed by rapid regrowth. Currently, little is known concerning the perception, signaling, or molecular responses associated with wound stress in forage- and turf-related grasses. A transcriptome analysis of Lolium temulentum plants subjected to severe wounding revealed 9413 upregulated and 7704 downregulated, distinct, differentially expressed genes (DEGs). Categories related to signaling, transcription, and response to stimuli were enriched in the upregulated DEGs. Specifically, sequences annotated as enzymes involved in hormone biosynthesis/action and cell wall modifications, mitogen-activated protein kinases, WRKY transcription factors, proteinase inhibitors, and pathogen defense-related DEGs were identified. Surprisingly, DEGs related to heat shock and chaperones were more prevalent in the downregulated DEGs when compared with the upregulated DEGs. This wound transcriptome analysis is the first step in identifying the molecular components and pathways used by grasses in response to wounding. The information gained from the analysis will provide a valuable molecular resource that will be used to develop approaches that can improve the recovery, regrowth, and long-term fitness of forage and turf grasses before/after cutting or grazing.
Comparative Functional Analysis of Class II Potassium Transporters, SvHKT2;1, SvHKT2;2, and HvHKT2;1, on Ionic Transport and Salt Tolerance in Transgenic Arabidopsis
Authors: Tada Yuichi , Ohnuma Aki
Published: June 2020

Class II high-affinity potassium transporters (HKT2s) mediate Na+–K+ cotransport and Na+/K+ homeostasis under K+-starved or saline conditions. Their functions have been studied in yeast and X. laevis oocytes; however, little is known about their respective properties in plant cells. In this study, we characterized the Na+ and K+ transport properties of SvHKT2;1, SvHKT2;2 and HvHKT2;1 in Arabidopsis under different ionic conditions. The differences were detected in shoot K+ accumulation and root K+ uptake under salt stress conditions, K+ accumulation in roots and phloem sap under K+-starved conditions, and shoot and root Na+ accumulation under K+-starved conditions among the HKT2s transgenic lines and WT plants. These results indicate the diverse ionic transport properties of these HKT2s in plant cells, which could not be detected using yeast or X. laevis oocytes. Furthermore, Arabidopsis expressing HKT2s showed reduced salt tolerance, while over-expression of HvHKT2;1 in barley, which has the ability to sequestrate Na+, showed enhanced salt tolerance by accumulating Na+ in the shoots. These results suggest that the coordinated enhancement of Na+ accumulation and sequestration mechanisms in shoots could be a promising strategy to confer salt tolerance to glycophytes.
Generation of High Yielding and Fragrant Rice (Oryza sativa L.) Lines by CRISPR/Cas9 Targeted Mutagenesis of Three Homoeologs of Cytochrome P450 Gene Family and OsBADH2 and Transcriptome and Proteome Profiling of Revealed Changes Triggered by Mutations
Authors: Usman Babar , Nawaz Gul , Zhao Neng , Liu Yaoguang , Li Rongbai
Published: June 2020

The significant increase in grain yield and quality are often antagonistic but a constant demand for breeders and consumers. Some genes related to cytochrome P450 family are known for rice organ growth but their role in controlling grain yield is still unknown. Here, we generated new rice mutants with high yield and improved aroma by simultaneously editing three cytochrome P450 homoeologs (Os03g0603100, Os03g0568400, and GL3.2) and OsBADH2 with the CRISPR/Cas9 system, and RNA-sequencing and proteomic analysis were performed to unveil the subsequent changes. High mutation efficiency was achieved in both target sites of each gene and the mutations were predominantly only deletions, while insertions were rare, and no mutations were detected in the five most likely off-target sites against each sgRNA. Mutants exhibited increased grain size, 2-acetyl-1-pyrroline (2AP) content, and grain cell numbers while there was no change in other agronomic traits. Transgene-DNA-free mutant lines appeared with a frequency of 44.44% and homozygous mutations were stably transmitted, and bi-allelic and heterozygous mutations followed Mendelian inheritance, while the inheritance of chimeric mutations was unpredictable. Deep RNA sequencing and proteomic results revealed the regulation of genes and proteins related to cytochrome P450 family, grain size and development, and cell cycle. The KEGG and hub-gene and protein network analysis showed that the gene and proteins related to ribosomal and photosynthesis pathways were mainly enriched, respectively. Our findings provide a broad and detailed basis to understand the role of CRISPR/Cas9 in rice yield and quality improvement.
Chemical and Enantioselective Analysis of the Leaf Essential Oil from Piper coruscans Kunth (Piperaceae), a Costal and Amazonian Native Species of Ecuador
Authors: Gilardoni Gianluca , Matute Yadira , Ramírez Jorge
Published: June 2020

In the present study, an essential oil was distilled from the leaves of Piper coruscans Kunth, a native Amazonian species belonging to the family Piperaceae and quite common in Ecuador. The chemical analysis was performed by GC-MS (qualitative) and GC-FID (quantitative), on polar and non-polar columns, detecting a total of 58 compounds of which 52 were identified. All the identified compounds were quantified. The essential oil was mainly constituted of sesquiterpenes (54.1–55.0%) and oxygenated sesquiterpenoids (32.5–33.6%), the major constituents being: (E)-β-caryophyllene (24.1–25.0%), α-humulene (11.6–12.0%), caryophyllene oxide (9.3–10.9%), linalool (4.5–5.2%), humulene epoxide II (3.6–4.1%), (E)-nerolidol (3.7–4.0%), α-copaene (3.7–3.9%), α-muurolol (3.4–3.7%), α-selinene (3.4–3.5%), β-selinene (3.1–3.3%), and one undetermined oxygenated sesquiterpenoid (3.1–3.3%). The aqueous phase (hydrolate) of the distillation process was also submitted to chemical analysis, showing linalool as the main organic compound in solution, with a concentration of 12.3–15.7 mg/100 mL. The essential oil was than analyzed for the enantiomeric distribution of its monoterpene constituents, affording the following enantiomeric excesses in two β-cyclodextrin-based enantioselective columns: (1S,5S)-(-)-α-pinene (60.0–69.6%), (1S,5S)-(-)-β-pinene (5.2–7.2%), (R)-(-)-α-phellandrene (72.5–78.2%), (R)-(+)-limonene (28.6%) and (R)-(-)-linalool (1.8–3.1%).
Hyper-Crosslinked Polymer Nanocomposites Containing Mesoporous Silica Nanoparticles with Enhanced Adsorption Towards Polar Dyes
Authors: Guerritore Marco , Castaldo Rachele , Silvestri Brigida , Avolio Roberto , Cocca Mariacristina , Errico Maria Emanuela , Avella Maurizio , Gentile Gennaro , Ambrogi Veronica
Published: June 2020

The development of new styrene-based hyper-crosslinked nanocomposites (HCLN) containing mesoporous silica nanoparticles (MSN) is reported here as a new strategy to obtain functional high surface area materials with an enhanced hydrophilic character. The HCLN composition, morphology and porous structure were analyzed using a multi-technique approach. The HCLN displayed a high surface area (above 1600 m2/g) and higher microporosity than the corresponding hyper-crosslinked neat resin. The enhanced adsorption properties of the HCLN towards polar organic dyes was demonstrated through the adsorption of a reactive dye, Remazol Brilliant Blue R (RB). In particular, the HCLN containing 5phr MSN showed the highest adsorption capacity of RB.
Assessment of the Mechanical and Thermal Properties of Injection-Molded Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)/Hydroxyapatite Nanoparticles Parts for Use in Bone Tissue Engineering
Authors: Ivorra-Martinez Juan , Quiles-Carrillo Luis , Boronat Teodomiro , Torres-Giner Sergio , A. Covas José
Published: June 2020

In the present study, poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] was reinforced with hydroxyapatite nanoparticles (nHA) to produce novel nanocomposites for potential uses in bone reconstruction. Contents of nHA in the 2.5–20 wt % range were incorporated into P(3HB-co-3HHx) by melt compounding and the resulting pellets were shaped into parts by injection molding. The addition of nHA improved the mechanical strength and the thermomechanical resistance of the microbial copolyester parts. In particular, the addition of 20 wt % of nHA increased the tensile (Et) and flexural (Ef) moduli by approximately 64% and 61%, respectively. At the highest contents, however, the nanoparticles tended to agglomerate, and the ductility, toughness, and thermal stability of the parts also declined. The P(3HB-co-3HHx) parts filled with nHA contents of up to 10 wt % matched more closely the mechanical properties of the native bone in terms of strength and ductility when compared with metal alloys and other biopolymers used in bone tissue engineering. This fact, in combination with their biocompatibility, enables the development of nanocomposite parts to be applied as low-stress implantable devices that can promote bone reconstruction and be reabsorbed into the human body.