bluuebunny/crossref_metadata_2025 huggingface dataset:
New Insight into Antimicrobial Compounds from Food and Marine-Sourced Carnobacterium Species through Phenotype and Genome Analyses
Authors: Begrem Simon , Ivaniuk Flora , Gigout-Chevalier Frédérique , Kolypczuk Laetitia , Bonnetot Sandrine , Leroi Françoise , Grovel Olivier , Delbarre-Ladrat Christine , Passerini Delphine
Published: July 2020

Carnobacterium maltaromaticum and Carnobacterium divergens, isolated from food products, are lactic acid bacteria known to produce active and efficient bacteriocins. Other species, particularly those originating from marine sources, are less studied. The aim of the study is to select promising strains with antimicrobial potential by combining genomic and phenotypic approaches on large datasets comprising 12 Carnobacterium species. The biosynthetic gene cluster (BGCs) diversity of 39 publicly available Carnobacterium spp. genomes revealed 67 BGCs, distributed according to the species and ecological niches. From zero to six BGCs were predicted per strain and classified into four classes: terpene, NRPS (non-ribosomal peptide synthetase), NRPS-PKS (hybrid non-ribosomal peptide synthetase-polyketide synthase), RiPP (ribosomally synthesized and post-translationally modified peptide). In parallel, the antimicrobial activity of 260 strains from seafood products was evaluated. Among the 60% of active strains, three genomes were sequenced and submitted to a dereplication process. C. inhibens MIP2551 produced a high amountof H2O2, probably thanks to the presence of four oxidase-encoding genes. C. maltaromaticum EBP3019 and SF668 strains were highly efficient against Listeria monocytogenes. A new extracellular 16 kDa unmodified bacteriocin in the EBP3019 strain and five different bacteriocins in SF668 were highlighted. In this study, the overview of antimicrobial BGC and inhibitory activities of Carnobacterium spp. allowed the prediction of potential innovative natural products that could be relevant for biotechnological applications.
Oral Primo-Colonizing Bacteria Modulate Inflammation and Gene Expression in Bronchial Epithelial Cells
Authors: Mathieu Elliot , MacPherson Chad W. , Belvis Jocelyn , Mathieu Olivier , Robert Véronique , Saint-Criq Vinciane , Langella Philippe , Tompkins Thomas A. , Thomas Muriel
Published: July 2020

The microbiota of the mouth disperses into the lungs, and both compartments share similar phyla. Considering the importance of the microbiota in the maturation of the immunity and physiology during the first days of life, we hypothesized that primo-colonizing bacteria of the oral cavity may induce immune responses in bronchial epithelial cells. Herein, we have isolated and characterized 57 strains of the buccal cavity of two human newborns. These strains belong to Streptococcus, Staphylococcus, Enterococcus, Rothia and Pantoea genera, with Streptococcus being the most represented. The strains were co-incubated with a bronchial epithelial cell line (BEAS-2B), and we established their impact on a panel of cytokines/chemokines and global changes in gene expression. The Staphylococcus strains, which appeared soon after birth, induced a high production of IL-8, suggesting they can trigger inflammation, whereas the Streptococcus strains were less associated with inflammation pathways. The genera Streptococcus, Enterococcus and Pantoea induced differential profiles of cytokine/chemokine/growth factor and set of genes associated with maturation of morphology. Altogether, our results demonstrate that the microorganisms, primo-colonizing the oral cavity, impact immunity and morphology of the lung epithelial cells, with specific effects depending on the phylogeny of the strains.
Geochemistry of Tourmaline from the Laodou Gold Deposit in the West Qinling Orogen, Central China: Implications for the Ore-Forming Process
Authors: Jin Xiaoye , Sui Jixiang
Published: July 2020

The Laodou gold deposit, located in the West Qinling Orogen of central China, is a newly recognized intrusion-related gold deposit. It consists of auriferous quartz-sulfide-tourmaline and minor quartz-stibnite veins that are structurally controlled by fault zones transecting the host quartz diorite porphyry. Two types of tourmaline were identified in this study: Type 1 tourmaline occurs as quartz-tourmaline nodules within the quartz diorite porphyry, whereas type 2 tourmaline occurs as quartz-sulfide-tourmaline veins in auriferous lodes. Here, we present a major and trace element analysis by electron microprobe and laser ablation inductively coupled plasma mass spectrometry on these two types of tourmaline. Both tourmaline types fall into the alkali group, and are classified under the schorl-dravite solid solution series. The substitutions of FeMg–1, FeAl–1, AlO((Fe, Mg)(OH)) –1, and X-site vacancyCa–1 are inferred by the variations of their major element compositions. Field and mineralogy observations suggest that type 1 tourmaline is a product of the late crystallization process of the quartz diorite porphyry, whereas type 2 tourmaline coexists with Au-bearing arsenopyrite and is crystallized from the ore-forming fluids. Their rare earth element compositions record the related magmatic hydrothermal evolution. The Co and Ni concentrations of the coexisting type 2 tourmaline and arsenopyrite define a regression line (correlation coefficient = 0.93) with an angular coefficient of 0.66, which represents the Co/Ni ratio of the tourmaline and arsenopyrite-precipitating fluids. This value is close to the Co/Ni ratios of the host quartz diorite porphyry, indicating a magma origin of the ore-forming fluids. The substitution of Al3+ by Fe3+ in both tourmaline types shows that type 1 tourmaline approaches the end member of povondraite whereas type 2 tourmaline occurs in opposite plots near the end member of Oxy-dravite, reflecting a more oxidizing environment for type 2 tourmaline formation. Moreover, the redox-sensitive V and Cr values of type 2 tourmaline are commonly 1–2 orders of magnitude higher than those of type 1 tourmaline, which also suggests that type 2 tourmaline forms from more oxidizing fluids. Combined with gold occurrence and fluid properties, we propose that the increasing of oxygen fugacity in the ore-forming fluids is a trigger of gold precipitation.
Alkylated Sesamol Derivatives as Potent Antioxidants
Authors: C. Palheta Ivanete , R. Ferreira Lanalice , K. L. Vale Joyce , P. P. Silva Osmarina , M. Herculano Anderson , R. H. M. Oliveira Karen , Neto Antonio M. J. Chaves , M. Campos Joaquín , B. R. Santos Cleydson , S. Borges Rosivaldo
Published: July 2020

Sesamol is a phenolic derivative. Its antioxidant activity is low than that of Trolox and depends on benzodioxole moiety. Thus, a molecular modification strategy through alkylation, inspired by natural and synthetic antioxidants, was studied by molecular modeling at the DFT/B3LYP level of theory by comparing the 6-31+G(d,p) and 6-311++G(2d,2p) basis sets. All proposed derivatives were compared to classical related antioxidants such as Trolox, t-butylated hydroxytoluene (BHT) and t-butylated hydroxyanisole (BHA). According to our results, molecular orbitals, single electron or hydrogen-atom transfers, spin density distributions, and alkyl substitutions at the ortho positions related to phenol moiety were found to be more effective than any other positions. The trimethylated derivative was more potent than Trolox. t-Butylated derivatives were stronger than all other alkylated derivatives and may be new alternative forms of modified antioxidants from natural products with applications in the chemical, pharmaceutical, and food industries.
Comparative Lipidomic Study of Human Milk from Different Lactation Stages and Milk Formulas
Authors: Hewelt-Belka Weronika , Garwolińska Dorota , Młynarczyk Michał , Kot-Wasik Agata
Published: July 2020

In this report, we present a detailed comparison of the lipid composition of human milk (HM) and formula milk (FM) targeting different lactation stages and infant age range. We studied HM samples collected from 26 Polish mothers from colostrum to 19 months of lactation, along with FM from seven brands available on the Polish market (infant formula, follow-on formula and growing-up formula). Lipid extracts were analysed using liquid chromatography coupled to high-resolution mass spectrometry (LC–Q-TOF–MS). We found that the lipid composition of FM deviates significantly from the HM lipid profile in terms of qualitative and quantitative differences. FM had contrasting lipid profiles mostly across brands and accordingly to the type of fat added but not specific to the target age range. The individual differences were dominant in HM; however, differences according to the lactation stage were also observed, especially between colostrum and HM collected in other lactation stages. Biologically and nutritionally important lipids, such as long-chain polyunsaturated fatty acids (LC-PUFAs) containing lipid species, sphingomyelines or ether analogues of glycerophosphoethanoloamines were detected in HM collected in all studied lactation stages. The observed differences concerned all the major HM lipid classes and highlight the importance of the detailed compositional studies of both HM and FM.
Biomarkers of Browning in Cold Exposed Siberian Adults
Authors: Efremova Agrafena , Colleluori Georgia , Thomsky Mikhail , Perugini Jessica , Protasoni Marina , Reguzzoni Marcella , Faragalli Andrea , Carle Flavia , Giordano Antonio , Cinti Saverio
Published: July 2020

Cold-exposure promotes energy expenditure by inducing brown adipose tissue (BAT) thermogenesis, which over time, is also sustained by browning, the appearance, or increase, of brown-like cells into white fat depots. Identification of circulating markers reflecting BAT activity and browning is crucial to study this phenomenon and its triggers, also holding possible implications for the therapy of obesity and metabolic diseases. Using RT-qPCR, we evaluated the peripheral blood mononuclear cells (PBMC) expression profile of regulators of BAT activity (CIDEA, PRDM16), white adipocytes browning (HOXC9 and SLC27A1), and fatty acid β-oxidation (CPT1A) in 150 Siberian healthy miners living at extremely cold temperatures compared to 29 healthy subjects living in thermoneutral conditions. Anthropometric parameters, glucose, and lipid profiles were also assessed. The cold-exposed group showed significantly lower weight, BMI, hip circumference, and PBMC expression of CIDEA, but higher expression of HOXC9 and higher circulating glucose compared to controls. Within the cold-exposed group, BMI, total cholesterol, and the atherogenic coefficient were lower in individuals exposed to low temperatures for a longer time. In conclusion, human PBMC expresses the brown adipocytes marker CIDEA and the browning marker HOXC9, which, varying according to cold-exposure, possibly reflect changes in BAT activation and white fat browning.
Trends and Disparities of Energy Intake and Macronutrient Composition in China: A Series of National Surveys, 1982–2012
Authors: Zhu Zhenni , Yang Xiaoguang , Fang Yuehui , Zhang Jian , Yang Zhenyu , Wang Zhu , Liu Ailing , He Li , Sun Jing , Lian Yiyao , Ding Gangqiang , He Yuna
Published: July 2020

Background: China’s diet transition might offer guidance to undeveloped countries on the way to prosperity. This report describes the trends and disparities in energy and macronutrient composition among Chinese adults, and between subpopulations. Methods: Data for the current study were obtained from the 1982, 1992, 2002, and 2010–2012 China National Nutrition Survey (CNNS) rounds, which were nationally representative cross-sectional surveys. We applied 24-h dietary recall and food weighing to assess dietary intake. Results: There were 204,877 participants aged 20 years or older included in the current analysis. From 1982 to 2012, the estimated energy intake declined from 2614.7 kcal to 2063.9 kcal. The trend in the estimated percentage of energy intake from fat showed a spike. It increased from 16.3% to 33.1% (1992 vs. 1982 difference, 7.6%; 95% CI 7.4% to 7.7%; 2002 vs. 1992 difference, 7.7%; 95% CI 7.6% to 7.9%; 2012 vs. 2002 difference, 1.6%; 95% CI 1.4% to 1.7%; p < 0.01 for trend). The trends coincided in all the subgroups (all p < 0.01 for trend) except for the subgroup of those educated over 15 years, whose percentage of energy intake from fat declined from 37.4% to 36.6% (2012 vs. 2002 difference, −0.8%; 95% CI −1.6% to 0.0%). The estimated percentage of energy intake from carbohydrates declined from 74.0% to 55.0%. The ranges of the estimated percentage of energy intake from fat, within population subgroups stratified by education level, area and Gross national product (GNP) level, were narrowed. Conclusions: Quick improvements in society and the economy effectively curbed undernutrition, but easily triggered overnutrition. Disparities persistently existed between different subpopulations, while the gaps would narrow if comprehensive efforts were made. Education might be a promising way to prevent overnutrition during prosperous progress. The low-social profile populations require specific interventions so as to avoid further disease burdens.
The Epstein-Barr Virus-Encoded EBNA1 Protein Activates the Bone Morphogenic Protein (BMP) Signalling Pathway to Promote Carcinoma Cell Migration
Authors: Bridgewater Hannah E. , Date Kathryn L. , O’Neil John D. , Hu Chunfang , Arrand John R. , Dawson Christopher W. , Young Lawrence S.
Published: July 2020

The Epstein-Barr virus (EBV)-encoded nuclear antigen 1 (EBNA1) protein is expressed in all virus-associated malignancies, where it performs an essential role in the maintenance, replication and transcription of the EBV genome. In recent years, it has become apparent that EBNA1 can also influence cellular gene transcription. Here, we demonstrate that EBNA1 is able to stimulate the expression of the Transforming growth factor-beta (TGFβ) superfamily member, bone morphogenic protein 2 (BMP2), with consequential activation of the BMP signalling pathway in carcinoma cell lines. We show that BMP pathway activation is associated with an increase in the migratory capacity of carcinoma cells, an effect that can be ablated by the BMP antagonist, Noggin. Gene expression profiling of authentic EBV-positive nasopharyngeal carcinoma (NPC) tumours revealed the consistent presence of BMP ligands, established BMP pathway effectors and putative target genes, constituting a prominent BMP “signature” in this virus-associated cancer. Our findings show that EBNA1 is the major viral-encoded protein responsible for activating the BMP signalling pathway in carcinoma cells and supports a role for this pathway in promoting cell migration and possibly, metastatic spread.
Molecular Characterization of Clistobothrium sp. Viable Plerocercoids in Fresh Longfin Inshore Squid (Doryteuthis pealeii) and Implications for Cephalopod Inspection
Authors: Guardone Lisa , Giusti Alice , Bilska-Zajac Ewa , Malandra Renato , Różycki Miroslaw , Armani Andrea
Published: July 2020

Cephalopods, an appreciated seafood product, are common hosts of marine cestodes. The aim of this work is to report visible alive plerocercoids in longfin inshore squid (Doryteuthis pealeii), a cephalopod species commercialized as fresh and whole in Italy. Seventy D. pealeii from the Northwest Atlantic (FAO area 21) were collected and visually inspected. In total, 18 plerocercoid larvae were found in the viscera of 10 host specimens (P: 14.3% 95% CI 7.1–24.7; MI: 1.8, MA: 0.26; range 1–4) and molecularly analyzed targeting the variable D2 region of the large subunit (LSU) rRNA gene and the cytochrome c oxidase subunit I (COI) gene. The molecular characterization allowed to identify all the plerocercoids as Clistobothrium sp., a cestode of the Phyllobothriidae family with Lamnidae sharks as definitive hosts, and cephalopods as second intermediate hosts. These findings represent the first molecular record of Clistobothrium sp. in D. pealeii, thus contributing to elucidate its poorly known life cycle. Even if not affecting consumer’s health, these visible parasites may represent a reason for disgust for consumers. Therefore, the results suggest that Food Business Operators should also check for the presence of these visible parasites during inspection and underline the importance of a correct consumers’ education.
Micro/Nanorobot: A Promising Targeted Drug Delivery System
Authors: Hu Mengyi , Ge Xuemei , Chen Xuan , Mao Wenwei , Qian Xiuping , Yuan Wei-En
Published: July 2020

Micro/nanorobot, as a research field, has attracted interest in recent years. It has great potential in medical treatment, as it can be applied in targeted drug delivery, surgical operation, disease diagnosis, etc. Differently from traditional drug delivery, which relies on blood circulation to reach the target, the designed micro/nanorobots can move autonomously, which makes it possible to deliver drugs to the hard-to-reach areas. Micro/nanorobots were driven by exogenous power (magnetic fields, light energy, acoustic fields, electric fields, etc.) or endogenous power (chemical reaction energy). Cell-based micro/nanorobots and DNA origami without autonomous movement ability were also introduced in this article. Although micro/nanorobots have excellent prospects, the current research is mainly based on in vitro experiments; in vivo research is still in its infancy. Further biological experiments are required to verify in vivo drug delivery effects of micro/nanorobots. This paper mainly discusses the research status, challenges, and future development of micro/nanorobots.
Avoiding the Pitfalls of siRNA Delivery to the Retinal Pigment Epithelium with Physiologically Relevant Cell Models
Authors: Ramsay Eva , Raviña Manuela , Sarkhel Sanjay , Hehir Sarah , Cameron Neil R. , Ilmarinen Tanja , Skottman Heli , Kjems Jørgen , Urtti Arto , Ruponen Marika , Subrizi Astrid
Published: July 2020

Inflammation is involved in the pathogenesis of several age-related ocular diseases, such as macular degeneration (AMD), diabetic retinopathy, and glaucoma. The delivery of anti-inflammatory siRNA to the retinal pigment epithelium (RPE) may become a promising therapeutic option for the treatment of inflammation, if the efficient delivery of siRNA to target cells is accomplished. Unfortunately, so far, the siRNA delivery system selection performed in dividing RPE cells in vitro has been a poor predictor of the in vivo efficacy. Our study evaluates the silencing efficiency of polyplexes, lipoplexes, and lipidoid-siRNA complexes in dividing RPE cells as well as in physiologically relevant RPE cell models. We find that RPE cell differentiation alters their endocytic activity and causes a decrease in the uptake of siRNA complexes. In addition, we determine that melanosomal sequestration is another significant and previously unexplored barrier to gene silencing in pigmented cells. In summary, this study highlights the importance of choosing a physiologically relevant RPE cell model for the selection of siRNA delivery systems. Such cell models are expected to enable the identification of carriers with a high probability of success in vivo, and thus propel the development of siRNA therapeutics for ocular disease.
Study on Ajuga reptans Extract: A Natural Antioxidant in Microencapsulated Powder Form as an Active Ingredient for Nutraceutical or Pharmaceutical Purposes
Authors: Esposito Tiziana , Sansone Francesca , Auriemma Giulia , Franceschelli Silvia , Pecoraro Michela , Picerno Patrizia , Aquino Rita P. , Mencherini Teresa
Published: July 2020

The administration of natural antioxidants is considered to be a prevention strategy for chronic diseases and a useful tool for the healthcare system to reduce the administration of expensive and often not effective treatments. The chemical characterization of a methanolic extract (AJ) of Ajuga reptans L. was performed, and its antioxidant activity was evaluated. AJ and the major compounds, characterized by chromatographic techniques as phenylpropanoids and iridoids, were able to reduce the Reactive Oxygen Species levels in cancer cell lines (melanoma, A375, cervical cancer, HeLa, and alveolar adenocarcinoma, A549), stimulated by E. coli lipopolysaccharide. However, a clinical translation of these results encountered a significant limitation represented by the poor water solubility and bioavailability of the extract and compounds. Consequently, a hydro-soluble powder system (AJEP3) was developed by spray-drying encapsulating AJ into a multi-component solid matrix that is based on L-proline and hydroxyethylcellulose as loading and coating agents, and lecithin as solubility enhancer. The technological approach led to a satisfactory process yield (71.5%), encapsulation efficiency (99.9%), and stability. The in vitro water dissolution rate of the bioactive compounds appeared to be improved with respect to the extract, suggesting higher feasibility in the manufacturing and administration; even the in vitro biological activity of the produced multi-component AJEP3 was clearly enhanced.
Y-Site Physical Compatibility of Numeta G13E with Drugs Frequently Used at Neonatal Intensive Care
Authors: Nezvalova-Henriksen Katerina , Nilsson Niklas , Østerberg Camilla Tomine , Staven Berge Vigdis , Tho Ingunn
Published: July 2020

Preterm neonates require parenteral nutrition (PN) in addition to intravenous drug therapy. Due to limited venous access, drugs are often co-administered with PN via the same lumen. If incompatible, precipitation and emulsion destabilization may occur with the consequent risk of embolism and hyper-immune reactions. Information on intravenous compatibility is scarce. Our aim was to analyse the compatibility of Numeta G13E with paracetamol, vancomycin and fentanyl because of the frequency of their use. A panel of methods was chosen to assess precipitation (sub-visual particle counting, turbidity measurement, Tyndall beam effect and pH measurement) and emulsion destabilization (mean droplet diameter measurement and sub-visual counting of oil droplets, followed by estimation of PFAT5 (percentage of fat residing in globules larger than 5 µm) and pH measurement). Samples in clinically relevant mixing ratios were tested immediately and after 4 h. All samples of drugs mixed with Numeta G13E were compared to unmixed controls. None of the tested drugs precipitated in contact with Numeta G13E, and we did not see any sign of emulsion destabilization when clinically relevant mixing ratios were applied. These results are reassuring. However, when contact time exceeds the established norm, caution in the form of filter utilisation and close inspection is advised.
DoE-Assisted Development of a Novel Glycosaminoglycan-Based Injectable Formulation for Viscosupplementation
Authors: Cicognani Marta , Rossi Silvia , Vecchi Gabriele , Giori Andrea Maria , Ferrari Franca
Published: July 2020

The aim of the present work was the development of a novel glycosaminoglycan (GAG)-based injectable formulation intended for intra-articular administration that should best mimic the healthy synovial fluid. Hyaluronic acid (HA) was chosen among GAG polymers, since it is the most abundant component of the synovial fluid. A DoE (Design of Experiment) approach was used for the development of a formulation containing two HA (very high (VHMW) and low (LMW) molecular weight) grades. The rationale for this choice is that so far, no commercial product based on a single HA grade or even on binary HA mixture possesses optimal viscoelastic properties in comparison with healthy synovial fluid. A full factorial design was chosen to investigate the influence of concentration and relative fraction of the two polymer grades (retained as factors of the model) on formulation functional (viscosity and viscoelastic) properties, which are considered response variables. Thanks to the DoE approach, the composition of the optimized HA formulation was found. The addition to such formulation of an injectable grade fat-free soy phospholipid, which was rich in phosphatidylcholine (PC), resulted in improved lubrication properties. The final HA + PC formulation, packaged in pre-filled sterile syringes, was stable in long-term and accelerated ICH (International Council for Harmonisation) storage conditions. The overall results pointed out the formulation suitability for further steps of pharmaceutical developments, namely for the passage to pilot scale.
Development and Characterization of Potential Ocular Mucoadhesive Nano Lipid Carriers Using Full Factorial Design
Authors: Kiss Eszter L. , Berkó Szilvia , Gácsi Attila , Kovács Anita , Katona Gábor , Soós Judit , Csányi Erzsébet , Gróf Ilona , Harazin András , Deli Mária A. , Balogh György T. , Budai-Szűcs Mária
Published: July 2020

Generally, topically applied eye drops have low bioavailability due to short residence time and low penetration of the drug. The aim of the present study was to incorporate dexamethasone (DXM) into nano lipid carriers (NLC), which contain mucoadhesive polymer, in order to increase the bioavailability of the drug. A 23 factorial experimental design was applied, in which the three factors were the polymer, the DXM, and the emulsifier concentrations. The samples were analyzed for particle size, zeta potential, polydispersity index, and Span value. The significant factors were identified. The biocompatibility of the formulations was evaluated with human corneal toxicity tests and immunoassay analysis. The possible increase in bioavailability was analyzed by means of mucoadhesivity, in vitro drug diffusion, and different penetration tests, such as in vitro cornea PAMPA model, human corneal cell penetration, and ex vivo porcine corneal penetration using Raman mapping. The results indicated that DXM can be incorporated in stable mucoadhesive NLC systems, which are non-toxic and do not have any harmful effect on cell junctions. Mucoadhesive NLCs can create a depot on the surface of the cornea, which can predict improved bioavailability.
PLGA Microspheres of hGH of Preserved Native State Prepared Using a Self-Regulated Process
Authors: Diana Jebun Nessa , Tao Ying , Du Qiran , Wang Meng , Kumar Chinta Uday , Wu Fei , Jin Tuo
Published: July 2020

The challenges of formulating recombinant human growth hormone (rhGH) into sustained-release polymeric microspheres include two mutual causal factors, protein denaturing by the formulation process and severe initial burst release related with relative high dose. The stabilizers to protect the proteins must not evoke osmotic pressure inside the microspheres, and the contact of the protein with the interface between water and organic solution of the polymer must be minimized. To meet these criteria, rhGH was pre-formulated into polysaccharide particles via an aqueous–aqueous emulsion in the present study, followed by encapsulating the particles into microspheres through a self-regulated process to minimize the contact of the protein with the water–oil interface. Polysaccharides as the protein stabilizer did not evoke osmotic pressure as small sugar stabilizers, the cause of severe initial burst release. Reduced initial burst enabled reduced protein loading to 9% (from 22% of the once commercialized Nutropin depot), which in turn reduced the dosage form index from 80 to 8.7 and eased the initial burst. A series of physical chemical characterizations as well as biologic and pharmacokinetic assays confirmed that the present method is practically feasible for preparing microspheres of proteins.
Combinatorial Inhibition of Cell Surface Receptors Using Dual Aptamer-Functionalized Nanoconstructs for Cancer Treatment
Authors: Lee Hyojin , Kim Tae Hee , Park Daechan , Jang Mihue , Chung Justin J. , Kim Soo Hyun , Kim Sang-Heon , Lee Kwan Hyi , Jung Youngmee , Oh Seung Ja
Published: July 2020

Membrane receptors overexpressed in diseased states are considered novel therapeutic targets. However, the single targeting approach faces several fundamental issues, such as poor efficacy, resistance, and toxicity. Here, we report a dual-targeting strategy to enhance anti-cancer efficacy via synergistic proximity interactions between therapeutics and two receptor proteins. Importantly, we report the first finding of an interaction between c-Met and nucleolin and demonstrate the therapeutic value of targeting the interaction between them. Bispecific nanocarriers densely grafted with anti-c-Met and -nucleolin aptamer increased the local concentration of aptamers at the target sites, in addition to inducing target receptor clustering. It was also demonstrated that the simultaneous targeting of c-Met and nucleolin inhibited the cellular functions of the receptors and increased anti-cancer efficacy by altering the cell cycle. Our findings pave the way for the development of an effective combinatorial treatment based on nanoconstruct-mediated interaction between receptors.
Orodispersible Polymer Films with the Poorly Water-Soluble Drug, Olanzapine: Hot-Melt Pneumatic Extrusion for Single-Process 3D Printing
Authors: Cho Hui-Won , Baek Seung-Hoon , Lee Beom-Jin , Jin Hyo-Eon
Published: July 2020

Amorphous solid dispersions (ASDs) improve the oral delivery of poorly water-soluble drugs. ASDs of olanzapine (OLZ), which have a high melting point and low solubility, are performed using a complicated process. Three-dimensional (3D) printing based on hot-melt pneumatic extrusion (HMPE) is a simplified method for producing ASDs. Unlike general 3D printing, printlet extrusion is possible without the preparation of drug-loaded filaments. By heating powder blends, direct fused deposition modeling (FDM) printing through a nozzle is possible, and this step produces ASDs of drugs. In this study, we developed orodispersible films (ODFs) loaded with OLZ as a poorly water-soluble drug. Various ratios of film-forming polymers and plasticizers were investigated to enhance the printability and optimize the printing temperature. Scanning electron microscopy (SEM) showed the surface morphology of the film for the optimization of the polymer carrier ratios. Differential scanning calorimetry (DSC) was used to evaluate thermal properties. Powder X-ray diffraction (PXRD) confirmed the physical form of the drug during printing. The 3D printed ODF formulations successfully loaded ASDs of OLZ using HMPE. Our ODFs showed fast disintegration patterns within 22 s, and rapidly dissolved and reached up to 88% dissolution within 5 min in the dissolution test. ODFs fabricated using HMPE in a single process of 3D printing increased the dissolution rates of the poorly water-soluble drug, which could be a suitable formulation for fast drug absorption. Moreover, this new technology showed prompt fabrication feasibility of various formulations and ASD formation of poorly water-soluble drugs as a single process. The immediate dissolution within a few minutes of ODFs with OLZ, an atypical antipsychotic, is preferred for drug compliance and administration convenience.
In Vitro Propagation of Isoëtes sabatina (Isoetaceae): A Key Conservation Challenge for a Critically Endangered Quillwort
Authors: Magrini Sara , Azzella Mattia M. , Bolpagni Rossano , Zucconi Laura
Published: July 2020

Isoëtes sabatina is an aquatic quillwort endemic to Italy. It is one of the rarest quillworts in Europe, and is critically endangered due to restricted range and to the continuous decline of both population and habitat quality. This study aims to develop an optimized protocol to reproduce and grow I. sabatina sporelings. Mature and immature megaspores were mixed with mature microspores to evaluate the influence of the developmental stage on germination and sporeling development. Two substrates, distilled water and water-agar medium, were tested for germination and sporeling emergence, and three substrates, sand, lake sediment and water-agar, were tested for transplants. A high percentage of megaspore germination (a total of 79.1%) was obtained in both substrates, higher for mature than immature spores. A total of 351 sporelings were produced in distilled water and water-agar cultures, with similar percentages (64.5% and 69.6%, respectively). The development stage of the megaspores affected both germination and sporeling development. Sporeling emergence showed significantly higher percentages in mature megaspores than immature ones (69.6% vs. 11.6%, respectively), with 85% of germinated spores developing sporelings. Only transplants over water-agar medium were successful. This protocol could be useful for the propagation of sporelings as the key step towards the planning of in situ actions to save this Mediterranean quillwort from extinction.
Identification and Expression Analysis of Cold Shock Protein 3 (BcCSP3) in Non-Heading Chinese Cabbage (Brassica rapa ssp. chinensis)
Authors: Huang Feiyi , Wang Jin , Duan Weike , Hou Xilin
Published: July 2020

A cold-related protein, cold shock protein 3 (BcCSP3), was isolated from non-heading Chinese cabbage in this study. BcCSP3 can encode 205 amino acids (aa) with an open reading frame (ORF) of 618 base pairs (bp). Multiple sequence alignment and phylogenetic tree analyses showed that BcCSP3 contains an N-terminal cold shock domain and is highly similar to AtCSP2, their kinship is recent. Real-time quantitative polymerase chain reaction (RT-qPCR) showed that the expression level of BcCSP3 in stems and leaves is higher than that in roots. Compared with other stress treatments, the change in BcCSP3 expression level was most pronounced under cold stress. In addition, a BcCSP3–GFP fusion protein was localized to the nucleus and cytoplasm. These results indicated that BcCSP3 may play an important role in response to cold stress in non-heading Chinese cabbage. This work may provide a reference for the identification and expression analysis of other CSP genes in non-heading Chinese cabbage.
RNAseq Analysis Reveals Altered Expression of Key Ion Transporters Causing Differential Uptake of Selective Ions in Canola (Brassica napus L.) Grown under NaCl Stress
Authors: Ulfat Mobina , Athar Habib-ur-Rehman , Khan Zaheerud-din , Kalaji Hazem M.
Published: July 2020

Salinity is one of the major abiotic stresses prevailing throughout the world that severely limits crop establishment and production. Every crop has an intra-specific genetic variation that enables it to cope with variable environmental conditions. Hence, this genetic variability is a good tool to exploit germplasms in salt-affected areas. Further, the selected cultivars can be effectively used by plant breeders and molecular biologists for the improvement of salinity tolerance. In the present study, it was planned to identify differential expression of genes associated with selective uptake of different ions under salt stress in selected salt-tolerant canola (Brassica napus L.) cultivar. For the purpose, an experiment was carried out to evaluate the growth response of different salt-sensitive and salt-tolerant canola cultivars. Plants were subjected to 200 mM NaCl stress. Canola cultivars—Faisal Canola, DGL, Dunkled, and CON-II—had higher growth than in cvs Cyclone, Ac-EXcel, Legend, and Oscar. Salt-tolerant cultivars were better able to maintain plant water status probably through osmotic adjustment as compared to salt-sensitive cultivars. Although salt stress increased shoot Na+ and shoot Cl− contents in all canola cultivars, salt-tolerant cultivars had a lower accumulation of these toxic nutrients. Similarly, salt stress reduced shoot K+ and Ca2+ contents in all canola cultivars, while salt-tolerant cultivars had a higher accumulation of K+ and Ca2+ in leaves, thereby having greater shoot K+/Na+ and Ca2+/Na+ ratios. Nutrient utilization efficiency decreased significantly in all canola cultivars due to the imposition of salt stress; however, it was greater in salt-tolerant cultivars—Faisal Canola, DGL, and Dunkled. Among four salt-tolerant canola cultivars, cv Dunkled was maximal in physiological attributes, and thus differentially expressed genes (DEGs) were assessed in it by RNA-seq analysis using next-generation sequencing (NGS) techniques. The differentially expressed genes (DEG) in cv Dunkled under salt stress were found to be involved in the regulation of ionic concentration, photosynthesis, antioxidants, and hormonal metabolism. However, the most prominent upregulated DEGs included Na/K transporter, HKT1, potassium transporter, potassium channel, chloride channel, cation exchanger, Ca channel. The RNA-seq data were validated through qRT-PCR. It was thus concluded that genes related to the regulation of ionic concentrate are significantly upregulated and expressed under salt stress, in the cultivar Dunkled.
Limited Nitrogen and Plant Growth Stages Discriminate Well Nitrogen Use, Uptake and Utilization Efficiency in Popcorn
Authors: Khan Shahid , Amaral Júnior Antônio Teixeira do , Ferreira Fernando Rafael Alves , Kamphorst Samuel Henrique , Gonçalves Gabriel Moreno Bernardo , Simone Mendonça Freitas Marta , Silveira Vanildo , Apolinário de Souza Filho Gonçalo , Francisco Teixeira do Amaral José , Enrique Bresssan Smith Ricardo , Hussain Khalil Iftikhar , Maganha Silva Vivas Janieli , Pequeno de Souza Yure , Alves Peçanha Diego
Published: July 2020

The extensive use of nitrogen (N) in agriculture has caused negative impacts on the environment and costs. In this context, two pot experiments were performed under different N levels and harvested at different vegetative stages to assess two popcorn inbred lines (P2 and L80) and their hybrid (F1 = P2 × L80) for the N use, uptake and utilization efficiency (with the inclusion and exclusion of root N content); to find the contrasting N levels and vegetative stages that effect nitrogen use efficiency (NUE) and to understand the relationship between the traits related to NUE. The hybrid and P2 were confirmed better than L80 for all the studied traits. NUE is mainly affected by the shoot dry weight, uptake and utilization efficiency. Extremely low and high N levels were found to be more discriminating for N use and dry weight, respectively. At the V6 (six fully expanded leaf) stage, root N content (RNC) should be considered; in contrast, at the VT (tasseling stage) stage, RNC should not be considered for the uptake and utilization efficiency. The genetic parameter performance for N use, uptake, shoot dry weight and N content could favor the achievement of the genetic gain in advanced segregating generations.
Feeding Behavior and Virus-transmission Ability of Insect Vectors Exposed to Systemic Insecticides
Authors: Garzo Elisa , Moreno Aránzazu , Plaza María , Fereres Alberto
Published: July 2020

The majority of plant viruses depend on Hemipteran vectors for their survival and spread. Effective management of these insect vectors is crucial to minimize the spread of vector-borne diseases, and to reduce crop damage. The aim of the present study was to evaluate the effect of various systemic insecticides on the feeding behavior of Bemisia tabaci and Myzus persicae, as well as their ability to interfere with the transmission of circulative viruses. The obtained results indicated that some systemic insecticides have antifeeding properties that disrupt virus transmission by their insect vectors. We found that some of the tested insecticides significantly reduced phloem contact and sap ingestion by aphids and whiteflies, activities that are closely linked to the transmission of phloem-limited viruses. These systemic insecticides may play an important role in reducing the primary and secondary spread of tomato yellow leaf curl virus (TYLCV) and turnip yellows virus (TuYV), transmitted by B. tabaci and M. persicae, respectively.
Advances in Plant Regeneration: Shake, Rattle and Roll
Authors: Ibáñez Sergio , Carneros Elena , Testillano Pilar S. , Pérez-Pérez José Manuel
Published: July 2020

Some plant cells are able to rebuild new organs after tissue damage or in response to definite stress treatments and/or exogenous hormone applications. Whole plants can develop through de novo organogenesis or somatic embryogenesis. Recent findings have enlarged our understanding of the molecular and cellular mechanisms required for tissue reprogramming during plant regeneration. Genetic analyses also suggest the key role of epigenetic regulation during de novo plant organogenesis. A deeper understanding of plant regeneration might help us to enhance tissue culture optimization, with multiple applications in plant micropropagation and green biotechnology. In this review, we will provide additional insights into the physiological and molecular framework of plant regeneration, including both direct and indirect de novo organ formation and somatic embryogenesis, and we will discuss the key role of intrinsic and extrinsic constraints for cell reprogramming during plant regeneration.
Russeting in ‘Apple’ Mango: Triggers and Mechanisms
Authors: Athoo Thomas O. , Winkler Andreas , Knoche Moritz
Published: July 2020

Russeting is an important surface disorder of many fruitcrop species. The mango cultivar ‘Apple’ is especially susceptible to russeting. Russeting compromises both fruit appearance and postharvest performance. The objective was to identify factors, mechanisms, and consequences of russeting in ‘Apple’ mango. Russeting was quantified on excised peels using image analysis and a categorical rating scheme. Water vapour loss was determined gravimetrically. The percentage of the skin area exhibiting russet increased during development. Russet began at lenticels then spread across the surface, ultimately forming a network of rough, brown patches over the skin. Cross-sections revealed stacks of phellem cells, typical of a periderm. Russet was more severe on the dorsal surface of the fruit than on the ventral and more for fruit in the upper part of the canopy than in the lower. Russet differed markedly across orchards sites of different climates. Russet was positively correlated with altitude, the number of rainy days, and the number of cold nights but negatively correlated with minimum, maximum, and mean daily temperatures, dew point temperature, and heat sum. Russeted fruit had higher transpiration rates than non-russeted fruits and higher skin permeance to water vapour. Russet in ‘Apple’ mango is due to periderm formation that is initiated at lenticels. Growing conditions conducive for surface wetness exacerbate russeting.
Response of Pumpkin to Different Concentrations and Forms of Selenium and Iodine, and their Combinations
Authors: Golob Aleksandra , Kroflič Ana , Jerše Ana , Kacjan Maršić Nina , Šircelj Helena , Stibilj Vekoslava , Germ Mateja
Published: July 2020

The elements selenium (Se) and iodine (I) are both crucial for the normal functioning of the thyroid. Biofortification with these elements is particularly feasible in areas where they show a deficit. Iodine and selenium can have positive effects on different plants when applied at the correct concentrations. The effects of their simultaneous addition on plant physiology and biochemistry, as well as on seed germination and sprout biomass, were studied in pumpkin (Cucurbita pepo L. ssp. pepo). To study the effect of Se and I on sprouts, sprouts were grown from seeds soaked in solutions of different forms of Se, I and their combination in the growth chamber experiment. In the field experiment, pumpkins plants were foliarly treated with the same concentrations and forms of Se and I. The combination of Se and I treatments enhanced the germination of the soaked seeds, with no significant differences between Se and I treatments for sprout mass. The yield of pumpkins and seed production were unaffected by Se and I foliar application. The anthocyanin levels and respiratory potential measured via the electron transport system’s activity showed different patterns according to treatments and plant parts (sprouts, leaves, seeds). The redistribution of Se and I from seeds to sprouts was significant. The accumulation of Se was higher in sprouts from the seeds treated with Se together with I, compared to sprouts from the seeds treated with Se alone. Interactions between Se and I were also noted in the seeds, which developed in the treated plants.
Coupling Phosphate-Solubilizing Bacteria with Phosphorus Supplements Improve Maize Phosphorus Acquisition and Growth under Lime Induced Salinity Stress
Authors: Adnan Muhammad , Fahad Shah , Zamin Muhammad , Shah Shahen , Mian Ishaq Ahmad , Danish Subhan , Zafar-ul-Hye Muhammad , Battaglia Martin Leonardo , Naz Raja Mohib Muazzam , Saeed Beena , Saud Shah , Ahmad Imran , Yue Zhen , Brtnicky Martin , Holatko Jiri , Datta Rahul
Published: July 2020

Global warming promotes soil calcification and salinization processes. As a result, soil phosphorus (P) is becoming deficient in arid and semiarid areas throughout the world. In this pot study, we evaluated the potential of phosphate-solubilizing bacteria (PSB) for enhancing the growth and P uptake in maize under varying levels of lime (4.8%, 10%, 15% and 20%) and additional P supplements (farmyard manure, poultry manure, single super phosphate and rock phosphate) added at the rate of 45 mg P2O5 kg−1. Inoculation and application of P as organic manures (Poultry and farm yard manures) improved maize growth and P uptake compared to the control and soils with P applied from mineral sources. Liming adversely affected crop growth, but the use of PSB and organic manure significantly neutralized this harmful effect. Mineral P sources combined with PSB were as effective as the organic sources alone. Furthermore, while single supper phosphate showed better results than Rock phosphate, the latter performed comparably upon PSB inoculation. Thus, PSB plus P application as organic manures is an eco-friendly option to improve crop growth and P nutrition in a calcareous soil under changing climate.
Selenium Enhances Cadmium Accumulation Capability in Two Mustard Family Species—Brassica napus and B. juncea
Authors: Zhang Zhong-Wei , Dong Yi-Ying , Feng Ling-Yang , Deng Zong-Lin , Xu Qiang , Tao Qi , Wang Chang-Quan , Chen Yang-Er , Yuan Ming , Yuan Shu
Published: July 2020

Oilseed rape (Brassica napus) is a Cadmium (Cd) hyperaccumulator. However, high-level Cd at the early seedling stage seriously arrests the growth of rape, which limits its applications. Brassica juncea had higher Cd accumulation capacity, but its biomass was lower, also limiting its applications. Previous studies have confirmed that Selenium (Se) can alleviate Cd toxicity. However, the regulatory mechanism of Se in different valence states of Cd accumulation was unclear. In this study, we investigated the ameliorating effects of three Se valence states, Na2SeO4 [Se(VI)], Na2SeO3 [Se(IV)] and Se-Met [Se(II)], to Cd toxicity by physiological and biochemical approaches in hydroponically-cultured Brassica juncea and Brassica napus seedlings. Although Se treatments slightly inhibited seedling Cd concentration, it tripled or quadrupled the Cd accumulation level per plant, because dry weight increased about four times more with Se and Cd application than with Cd treatment alone. Among the different valence states of Se, Se(II) had the most marked effect on reducing Cd toxicity as evidenced by decreased growth inhibition and Cd content. The application of Se(II) was effective in reducing Cd-induced reactive oxygen species accumulation, and promoted the antioxidant enzyme activity and photosynthesis of both Brassica species. In addition, Se(II) treatment increased the concentrations of Cd in the cell wall and soluble fractions, but the Cd concentration in the organelle part was reduced.