bluuebunny/crossref_metadata_2025 huggingface dataset:
Aflatoxins: Occurrence, Exposure, and Binding to Lactobacillus Species from the Gut Microbiota of Rural Ugandan Children
Authors: Wacoo Alex Paul , Atukunda Prudence , Muhoozi Grace , Braster Martin , Wagner Marijke , van den Broek Tim J , Sybesma Wilbert , Westerberg Ane C. , Iversen Per Ole , Kort Remco
Published: March 2020

Chronic exposure of children in sub-Saharan Africa to aflatoxins has been associated with low birth weight, stunted growth, immune suppression, and liver function damage. Lactobacillus species have been shown to reduce aflatoxin contamination during the process of food fermentation. Twenty-three Lactobacillus strains were isolated from fecal samples obtained from a cohort of rural Ugandan children at the age of 54 to 60 months, typed by 16S rRNA gene sequencing, and characterized in terms of their ability to bind aflatoxin B1 in vitro. Evidence for chronic exposure of these children to aflatoxin B1 in the study area was obtained by analysis of local foods (maize flour and peanuts), followed by the identification of the breakdown product aflatoxin M1 in their urine samples. Surprisingly, Lactobacillus in the gut microbiota of 140 children from the same cohort at 24 and 36 months showed the highest positive correlation coefficient with stunting among all bacterial genera identified in the stool samples. This correlation was interpreted to be associated with dietary changes from breastfeeding to plant-based solid foods that pose an additional risk for aflatoxin contamination, on one hand, and lead to increased intake of Lactobacillus species on the other.
The Importance of Transaminases Flare in Liver Elastography: Characterization of the Probability of Liver Fibrosis Overestimation by Hepatitis C Virus-Induced Cytolysis
Authors: Giuffrè Mauro , Fouraki Sofia , Comar Manola , Masutti Flora , Crocè Lory Saveria
Published: March 2020

Background: Liver stiffness measurement (LSM) is crucial for appropriate fibrosis staging in patients with ongoing hepatitis C virus (HCV) infection. However, there is still an ongoing debate on the impact of serum transaminases (aspartate-aminotransferase, AST; alanine-aminotransferase, ALT) on LSM. Methods: We selected 110 patients undergoing HCV eradication therapy with LSM compatible with significant liver fibrosis. LSM was evaluated prior to therapy and one year after HCV eradication. Results: LSM showed a median decrease of 35% from baseline values, and 67 (61%) patients showed posttreatment values compatible with lower fibrosis stages. We developed two logistic regression models to determine the probability of liver fibrosis overestimation according to serum transaminase. The probability of overestimation of two or more fibrosis grade is equal to (1) 50% for AST of 99 IU/L (2.2 ULN) and ALT of 90.5 IU/L (2 ULN), (2) 80% for AST of 123.5 IU/L (2.74 ULN) and ALT of 101.5 IU/L (2.25 ULN), and (3) reaches 100% for AST of 211 IU/L (4.7 ULN) and ALT of 140 IU/L (3.1 ULN). Conclusions: This study highlights the impact of serum transaminases on LSM. We believe that our findings may serve as a reference point for appropriate fibrosis stratification by liver elastography in patients with HCV infection.
Isoprene Oxidation by the Gram-Negative Model bacterium Variovorax sp. WS11
Authors: Dawson Robin A. , Larke-Mejía Nasmille L. , Crombie Andrew T. , Ul Haque Muhammad Farhan , Murrell J. Colin
Published: March 2020

Plant-produced isoprene (2-methyl-1,3-butadiene) represents a significant portion of global volatile organic compound production, equaled only by methane. A metabolic pathway for the degradation of isoprene was first described for the Gram-positive bacterium Rhodococcus sp. AD45, and an alternative model organism has yet to be characterised. Here, we report the characterisation of a novel Gram-negative isoprene-degrading bacterium, Variovorax sp. WS11. Isoprene metabolism in this bacterium involves a plasmid-encoded iso metabolic gene cluster which differs from that found in Rhodococcus sp. AD45 in terms of organisation and regulation. Expression of iso metabolic genes is significantly upregulated by both isoprene and epoxyisoprene. The enzyme responsible for the initial oxidation of isoprene, isoprene monooxygenase, oxidises a wide range of alkene substrates in a manner which is strongly influenced by the presence of alkyl side-chains and differs from other well-characterised soluble diiron monooxygenases according to its response to alkyne inhibitors. This study presents Variovorax sp. WS11 as both a comparative and contrasting model organism for the study of isoprene metabolism in bacteria, aiding our understanding of the conservation of this biochemical pathway across diverse ecological niches.
New Antimicrobial Phenyl Alkenoic Acids Isolated from an Oil Palm Rhizosphere-Associated Actinomycete, Streptomyces palmae CMU-AB204T
Authors: Sujarit Kanaporn , Mori Mihoko , Dobashi Kazuyuki , Shiomi Kazuro , Pathom-aree Wasu , Lumyong Saisamorn
Published: March 2020

Basal stem rot (BSR), or Ganoderma rot disease, is the most serious disease associated with the oil palm plant of Southeast Asian countries. A basidiomycetous fungus, Ganoderma boninense, is the causative microbe of this disease. To control BSR in oil palm plantations, biological control agents are gaining attention as a major alternative to chemical fungicides. In the course of searching for effective actinomycetes as potential biological control agents for BSR, Streptomyces palmae CMU-AB204T was isolated from oil palm rhizosphere soil collected on the campus of Chiang Mai University. The culture broth of this strain showed significant antimicrobial activities against several bacteria and phytopathogenic fungi including G. boninense. Antifungal and antibacterial compounds were isolated by antimicrobial activity-guided purification using chromatographic methods. Their structures were elucidated by spectroscopic techniques, including Nuclear Magnetic Resonance (NMR), Mass Spectrometry (MS), Ultraviolet (UV), and Infrared (IR) analyses. The current study isolated new phenyl alkenoic acids 1–6 and three known compounds, anguinomycin A (7), leptomycin A (8), and actinopyrone A (9) as antimicrobial agents. Compounds 1 and 2 displayed broad antifungal activity, though they did not show antibacterial activity. Compounds 3 and 4 revealed a strong antibacterial activity against both Gram-positive and Gram-negative bacteria including the phytopathogenic strain Xanthomonas campestris pv. oryzae. Compounds 7–9 displayed antifungal activity against Ganoderma. Thus, the antifungal compounds obtained in this study may play a role in protecting oil palm plants from Ganoderma infection with the strain S. palmae CMU-AB204T.
A Streptomyces sp. NEAU-HV9: Isolation, Identification, and Potential as a Biocontrol Agent against Ralstonia solanacearum of Tomato Plants
Authors: Ling Ling , Han Xiaoyang , Li Xiao , Zhang Xue , Wang Han , Zhang Lida , Cao Peng , Wu Yutong , Wang Xiangjing , Zhao Junwei , Xiang Wensheng
Published: March 2020

Ralstonia solanacearum is an important soil-borne bacterial plant pathogen. In this study, an actinomycete strain named NEAU-HV9 that showed strong antibacterial activity against Ralstonia solanacearum was isolated from soil using an in vitro screening technique. Based on physiological and morphological characteristics and 98.90% of 16S rRNA gene sequence similarity with Streptomyces panaciradicis 1MR-8T, the strain was identified as a member of the genus Streptomyces. Tomato seedling and pot culture experiments showed that after pre-inoculation with the strain NEAU-HV9, the disease occurrence of tomato seedlings was effectively prevented for R. solanacearum. Then, a bioactivity-guided approach was employed to isolate and determine the chemical identity of bioactive constituents with antibacterial activity from strain NEAU-HV9. The structure of the antibacterial metabolite was determined as actinomycin D on the basis of extensive spectroscopic analysis. To our knowledge, this is the first report that actinomycin D has strong antibacterial activity against R. solanacearum with a MIC (minimum inhibitory concentration) of 0.6 mg L−1 (0.48 μmol L−1). The in vivo antibacterial activity experiment showed that actinomycin D possessed significant preventive efficacy against R. solanacearum in tomato seedlings. Thus, strain NEAU-HV9 could be used as BCA (biological control agent) against R. solanacearum, and actinomycin D might be a promising candidate for a new antibacterial agent against R. solanacearum.
RHΔgra17Δnpt1 Strain of Toxoplasma gondii Elicits Protective Immunity Against Acute, Chronic and Congenital Toxoplasmosis in Mice
Authors: Liang Qin-Li , Sun Li-Xiu , Elsheikha Hany M. , Cao Xue-Zhen , Nie Lan-Bi , Li Ting-Ting , Li Tao-Shan , Zhu Xing-Quan , Wang Jin-Lei
Published: March 2020

In the present study, a dense granule protein 17 (gra17) and novel putative transporter (npt1) double deletion mutant of Toxoplasma gondii RH strain was engineered. The protective efficacy of vaccination using RHΔgra17Δnpt1 tachyzoites against acute, chronic, and congenital toxoplasmosis was studied in a mouse model. Immunization using RHΔgra17Δnpt1 induced a strong humoral and cellular response, as indicated by the increased levels of anti-T. gondii specific IgG, interleukin 2 (IL-2), IL-10, IL-12, and interferon-gamma (IFN-γ). Vaccinated mice were protected against a lethal challenge dose (103 tachyzoites) of wild-type homologous (RH) strain and heterologous (PYS and TgC7) strains, as well as against 100 tissue cysts or oocysts of Pru strain. Vaccination also conferred protection against chronic infection with 10 tissue cysts or oocysts of Pru strain, where the numbers of brain cysts in the vaccinated mice were significantly reduced compared to those detected in the control (unvaccinated + infected) mice. In addition, vaccination protected against congenital infection with 10 T. gondii Pru oocysts (administered orally on day 5 of gestation) as shown by the increased litter size, survival rate and the bodyweight of pups born to vaccinated dams compared to those born to unvaccinated + infected dams. The brain cyst burden of vaccinated dams was significantly lower than that of unvaccinated dams infected with oocysts. Our data show that T. gondii RHΔgra17Δnpt1 mutant strain can protect mice against acute, chronic, and congenital toxoplasmosis by balancing inflammatory response with immunogenicity.
Acetate Production from Cafeteria Wastes and Corn Stover Using a Thermophilic Anaerobic Consortium: A Prelude Study for the Use of Acetate for the Production of Value-Added Products
Authors: David Aditi , Tripathi Abhilash Kumar , Sani Rajesh Kumar
Published: March 2020

Efficient and sustainable biochemical production using low-cost waste assumes considerable industrial and ecological importance. Solid organic wastes (SOWs) are inexpensive, abundantly available resources and their bioconversion to volatile fatty acids, especially acetate, aids in relieving the requirements of pure sugars for microbial biochemical productions in industries. Acetate production from SOW that utilizes the organic carbon of these wastes is used as an efficient solid waste reduction strategy if the environmental factors are optimized. This study screens and optimizes influential factors (physical and chemical) for acetate production by a thermophilic acetogenic consortium using two SOWs—cafeteria wastes and corn stover. The screening experiment revealed significant effects of temperature, bromoethane sulfonate, and shaking on acetate production. Temperature, medium pH, and C:N ratio were further optimized using statistical optimization with response surface methodology. The maximum acetate concentration of 8061 mg L−1 (>200% improvement) was achieved at temperature, pH, and C:N ratio of 60 °C, 6, 25, respectively, and acetate accounted for more than 85% of metabolites. This study also demonstrated the feasibility of using acetate-rich fermentate (obtained from SOWs) as a substrate for the growth of industrially relevant yeast Yarrowia lipolytica, which can convert acetate into higher-value biochemicals.
Phosphorus Inactivation in Lake Sediments Using Calcite Materials and Controlled Resuspension—Mechanism and Efficiency
Authors: Bańkowska-Sobczak Agnieszka , Blazejczyk Aurelia , Eiche Elisabeth , Fischer Uwe , Popek Zbigniew
Published: March 2020

The efficiency and mechanism of orthophosphate—soluble reactive phosphorus (SRP)—inactivation in eutrophic lakes using controlled resuspension and calcite application into the sediment were investigated in this study. Two calcite materials, industrially produced precipitated calcium carbonate (PCC) and natural ground limestone (GCC), were tested in short-term batch experiments and long-term sediment incubations under oxic and anoxic conditions. Maximum SRP adsorption capacity calculated using Langmuir model for PCC (3.11 mg PO43− g−1) was 6 times higher than of GCC (0.43 mg PO43− g−1), reflecting substantial difference in the surface area of calcite materials (12.36 and 1.72 m2 g−1, respectively). PCC applied into the sediment during controlled resuspension reduced SRP release by 95% (oxic) and 78% (anoxic incubation) at medium dose (0.75 kg m−2) and suppressed it completely at high dose (1.5 kg m−2) for at least 3 months, irrespectively of incubation conditions. The maximum achieved reduction of SRP release using GCC was also meaningful: 78% under oxic and 56% under anoxic conditions, but this required very high doses of this material (6 kg m−2). Mechanisms of SRP inactivation by calcites were: (1) adsorption of SRP during application into the resuspended sediment and (2) precipitation of calcium-phosphate compounds (Ca-PO4) during subsequent incubation, which was reflected in a substantial increase in the HCl-P fraction (phosphorus extractable in 0.5 M HCl) in sediments enriched with calcite, irrespectively of oxygen presence. However, anoxia strongly promoted the formation of this fraction: the rise of HCl-P was 2–6 times higher in anoxic than in oxic conditions, depending on the dose and form of calcite applied. The results showed that SRP inactivation using the controlled resuspension method is only successful if highly efficient reactive materials are used, due to large amount of SRP being released from sediment during resuspension. Thus, calcite materials exhibiting high adsorption capacity should be used in this lakes’ restoration technology to ensure fast and sufficient SRP inactivation. The rise in the HCl-P fraction in sediment suggests SRP inactivation through precipitation of relatively stable Ca-PO4 minerals, which makes calcite a suitable agent for sustainable, long term SRP inactivation. As anoxic conditions promoted formation of these compounds, calcite seems to be a promising SRP inactivation agent in highly reductive sediments.
Combination of MRI and SEM to Assess Changes in the Chemical Properties and Permeability of Porous Media due to Barite Precipitation
Authors: Poonoosamy Jenna , Haber-Pohlmeier Sabina , Deng Hang , Deissmann Guido , Klinkenberg Martina , Gizatullin Bulat , Stapf Siegfried , Brandt Felix , Bosbach Dirk , Pohlmeier Andreas
Published: March 2020

The understanding of the dissolution and precipitation of minerals and its impact on the transport of fluids in porous media is essential for various subsurface applications, including shale gas production using hydraulic fracturing (“fracking”), CO2 sequestration, or geothermal energy extraction. In this work, we conducted a flow through column experiment to investigate the effect of barite precipitation following the dissolution of celestine and consequential permeability changes. These processes were assessed by a combination of 3D non-invasive magnetic resonance imaging, scanning electron microscopy, and conventional permeability measurements. The formation of barite overgrowths on the surface of celestine manifested in a reduced transverse relaxation time due to its higher magnetic susceptibility compared to the original celestine. Two empirical nuclear magnetic resonance (NMR) porosity–permeability relations could successfully predict the observed changes in permeability by the change in the transverse relaxation times and porosity. Based on the observation that the advancement of the reaction front follows the square root of time, and micro-continuum reactive transport modelling of the solid/fluid interface, it can be inferred that the mineral overgrowth is porous and allows the diffusion of solutes, thus affecting the mineral reactivity in the system. Our current investigation indicates that the porosity of the newly formed precipitate and consequently its diffusion properties depend on the supersaturation in solution that prevails during precipitation.
Interfacial Energetic Level Mapping and Nano-Ordering of Small Molecule/Fullerene Organic Solar Cells by Scanning Tunneling Microscopy and Spectroscopy
Authors: Caballero-Quintana Irving , Romero-Borja Daniel , Maldonado José-Luis , Nicasio-Collazo Juan , Amargós-Reyes Olivia , Jiménez-González Antonio
Published: March 2020

Using scanning tunneling microscopy (STM) and spectroscopy (STS) at the liquid/solid interface, morphology evolution process and energetic level alignment of very thin solid films (thickness: <700 pm), of the low molecular weight molecule DRCN5T and DRCN5T:[70]PCBM blend are analyzed after applying thermal annealing at different temperatures. These films exhibit a worm-like pattern without thermal annealing (amorphous shape); however, after applying thermal annealing at 120 °C, the small molecule film domains crystallize verified by X-ray diffraction: structural geometry becomes a well-defined organized array. By using STS, the energy band diagrams of the semiconductor bulk heterojunction (blended film) at the donor-acceptor interface are determined; morphology and energy characteristics can be correlated with the organic solar cells (OSC) performance. When combining thermal treatment and solvent vapor annealing processes as described in previous literature by using other techniques, OSC devices based on DRCN5T show a very acceptable power conversion efficiency of 9.0%.
Electroactive 3D Printed Scaffolds Based on Percolated Composites of Polycaprolactone with Thermally Reduced Graphene Oxide for Antibacterial and Tissue Engineering Applications
Authors: Angulo-Pineda Carolina , Srirussamee Kasama , Palma Patricia , Fuenzalida Victor M. , Cartmell Sarah H. , Palza Humberto
Published: March 2020

Applying electrical stimulation (ES) could affect different cellular mechanisms, thereby producing a bactericidal effect and an increase in human cell viability. Despite its relevance, this bioelectric effect has been barely reported in percolated conductive biopolymers. In this context, electroactive polycaprolactone (PCL) scaffolds with conductive Thermally Reduced Graphene Oxide (TrGO) nanoparticles were obtained by a 3D printing method. Under direct current (DC) along the percolated scaffolds, a strong antibacterial effect was observed, which completely eradicated S. aureus on the surface of scaffolds. Notably, the same ES regime also produced a four-fold increase in the viability of human mesenchymal stem cells attached to the 3D conductive PCL/TrGO scaffold compared with the pure PCL scaffold. These results have widened the design of novel electroactive composite polymers that could both eliminate the bacteria adhered to the scaffold and increase human cell viability, which have great potential in tissue engineering applications.
Mineral-Doped Poly(L-lactide) Acid Scaffolds Enriched with Exosomes Improve Osteogenic Commitment of Human Adipose-Derived Mesenchymal Stem Cells
Authors: Gandolfi Maria Giovanna , Gardin Chiara , Zamparini Fausto , Ferroni Letizia , Esposti Micaela Degli , Parchi Greta , Ercan Batur , Manzoli Lucia , Fava Fabio , Fabbri Paola , Prati Carlo , Zavan Barbara
Published: March 2020

Exosomes derived from mesenchymal stem cells are extracellular vesicles released to facilitate cell communication and function. Recently, polylactic acid (PLA), calcium silicates (CaSi), and dicalcium phosphate dihydrate (DCPD) have been used to produce bioresorbable functional mineral-doped porous scaffolds-through thermally induced phase separation technique, as materials for bone regeneration. The aim of this study was to investigate the effect of mineral-doped PLA-based porous scaffolds enriched with exosome vesicles (EVs) on osteogenic commitment of human adipose mesenchymal stem cells (hAD-MSCs). Two different mineral-doped scaffolds were produced: PLA-10CaSi-10DCPD and PLA-5CaSi-5DCPD. Scaffolds surface micromorphology was investigated by ESEM-EDX before and after 28 days immersion in simulated body fluid (HBSS). Exosomes were deposited on the surface of the scaffolds and the effect of exosome-enriched scaffolds on osteogenic commitment of hAD-MSCs cultured in proximity of the scaffolds has been evaluated by real time PCR. In addition, the biocompatibility was evaluated by direct-contact seeding hAD-MSCs on scaffolds surface-using MTT viability test. In both formulations, ESEM showed pores similar in shape (circular and elliptic) and size (from 10–30 µm diameter). The porosity of the scaffolds decreased after 28 days immersion in simulated body fluid. Mineral-doped scaffolds showed a dynamic surface and created a suitable bone-forming microenvironment. The presence of the mineral fillers increased the osteogenic commitment of hAD-MSCs. Exosomes were easily entrapped on the surface of the scaffolds and their presence improved gene expression of major markers of osteogenesis such as collagen type I, osteopontin, osteonectin, osteocalcin. The experimental scaffolds enriched with exosomes, in particular PLA-10CaSi-10DCPD, increased the osteogenic commitment of MSCs. In conclusion, the enrichment of bioresorbable functional scaffolds with exosomes is confirmed as a potential strategy to improve bone regeneration procedures.
PL Tunable GaN Nanoparticles Synthesis through Femtosecond Pulsed Laser Ablation in Different Environments
Authors: Hao Juan , Xu Sijia , Gao Bingrong , Pan Lingyun
Published: March 2020

The tunable photoluminescence (PL) property is very important for gallium nitride (GaN) nanoparticles in the application of ultraviolet and blue optoelectronic devices, while conventional methods are not so satisfactory that alternative methods for preparing GaN nanoparticles should be studied. In this paper, ultra-small and well dispersed GaN nanoparticles are fabricated through femtosecond pulse laser ablation in air, water and ethanol. For the PL spectra of GaN nanoparticles, there are no shifts in air, red shifts in water and blue shifts in ethanol compared with the intrinsic PL spectra of bulk GaN. The X-ray photoelectron spectroscopy (XPS) results demonstrate that the various PL spectra can be due to the different components inside the GaN nanoparticles, which not only have effect on the PL emissions, but also greatly influence the intensity of PL. This study validates that the ablation environment has a great adjustable effect on the properties of GaN nanoparticles.
Hyperthermia Induced by Near-Infrared Laser-Irradiated CsWO3 Nanoparticles Disintegrates Preformed Lysozyme Amyloid Fibrils
Authors: Hu Po-Sheng , Tomasovicova Natalia , Chou Hsiu-Jen , Li Meng-Chang , Vojtko Marek , Zakutanska Katarina , Majorosova Jozefina , Chen Shean-Jen , Kopcansky Peter
Published: March 2020

This research study attempts to prove the concept of the applicability of hyperthermia to treating the lysozyme amyloid fibrils (LAFs)’s self-assembled fibrillary aggregates by a feedback-modulated temperature controller ranging from 26 °C to 80 °C, and separately, by near-infrared (NIR) laser-irradiated cesium tungstate (CsWO3) nanoparticle (NPs). The dependence of the final morphology of the amyloidal assembly on external heating and the photothermal effect of the NPs on treating the fibrillary assembly were investigated and analyzed. Experimentally, atomic force microscopy (AFM), optical stereoscopy, and scanning electron microscopy (SEM) were used primarily to ensure mutual interaction between LAFs and NPs, optically elucidate the surface contour and final fibrillary assembly upon the influence of thermal treatment, and further reveal fine-details of the optical samples. Finally, conclusive remarks are drawn that the fibrillary structures doped with the NPs exhibit an increasing degree of unique orthogonality. As the temperature rises, utter deformation of the dendritic structures of fibrillary assemblies at 70 °C was found, and NIR laser-irradiated CsWO3 NPs have been demonstrated to be useful in topically destructing pre-assembled LAFs, which may be conducive to the future development of neurodegenerative therapeutic techniques.
Ultrathin Washcoat and Very Low Loading Monolithic Catalyst with Outstanding Activity and Stability in Dry Reforming of Methane
Authors: Agueniou Fazia , Vidal Hilario , Yeste María Pilar , Hernández-Garrido Juan C. , Cauqui Miguel A. , Rodríguez-Izquierdo José M. , Calvino José J. , Gatica José M.
Published: March 2020

A Ni/CeO2/ZrO2 catalyst with improved redox properties has been washcoated onto a honeycomb cordierite monolith in the form of a nonconventional alumina-catalyst layer, just a few nanometers thick. In spite of the very low active phase loading, the monolith depicts outstanding performance in dry reforming of methane, both in terms of activity, with values reaching the thermodynamic limit already at 750 °C, even under extreme Weight Hourly Space Velocities (WHSV 115–346 L·g−1·h−1), as well as in terms of stability during prolonged Time on Stream (TOS 24–48 h).
Repositioning the Sm-Binding Site in Saccharomyces cerevisiae Telomerase RNA Reveals RNP Organizational Flexibility and Sm-Directed 3′-End Formation
Authors: Hass Evan P. , Zappulla David C.
Published: March 2020

Telomerase RNA contains a template for synthesizing telomeric DNA and has been proposed to act as a flexible scaffold for holoenzyme protein subunits in the RNP. In Saccharomyces cerevisiae, the telomerase RNA, TLC1, is bound by the Sm7 protein complex, which is required for stabilization of the predominant, non-polyadenylated (poly(A)–) TLC1 isoform. However, it remains unclear (1) whether Sm7 retains this function when its binding site is repositioned within TLC1, as has been shown for other TLC1-binding telomerase subunits, and (2) how Sm7 stabilizes poly(A)– TLC1. Here, we first show that Sm7 can stabilize poly(A)– TLC1 even when its binding site is repositioned via circular permutation to several different positions within TLC1, further supporting the conclusion that the telomerase holoenzyme is organizationally flexible. Next, we show that when an Sm site is inserted 5′ of its native position and the native site is mutated, Sm7 stabilizes shorter forms of poly(A)– TLC1 in a manner corresponding to how far upstream the new site was inserted, providing strong evidence that Sm7 binding to TLC1 controls where the mature poly(A)– 3′ is formed by directing a 3′-to-5′ processing mechanism. In summary, our results show that Sm7 and the 3′ end of yeast telomerase RNA comprise an organizationally flexible module within the telomerase RNP and provide insights into the mechanistic role of Sm7 in telomerase RNA biogenesis.
Using the Beth–Uhlenbeck Approach to Describe the Kaon to Pion Ratio in a 2 + 1 Flavor PNJL Model
Authors: Blaschke David , Friesen Alexandra , Kalinovsky Yuri , Radzhabov Andrey
Published: March 2020

The kaon to pion ratios are discussed in the framework of a 2 + 1 flavor PNJL model. In order to interpret the behavior of bound states in medium, the Beth–Uhlenbeck approach is used. It is shown that, in terms of phase shifts in the K + channel, an additional low-energy mode could appear as a bound state in medium, since the masses of the quark constituents are different. The comparison with experimental data for the ratios is performed and the influence of the anomalous mode to the “horn” effect in the K + / π + ratio is discussed.
NIR Laser-Responsive PNIPAM and Gold Nanorod Composites for the Engineering of Thermally Reactive Drug Delivery Nanomedicine
Authors: Kwon Yejin , Choi Yonghyun , Jang Jaehee , Yoon Semi , Choi Jonghoon
Published: March 2020

When ingesting a drug on its own or injecting it directly into tissue, its concentration increases immediately within the body, which often exacerbates the side effects and increases its toxicity. To solve this problem, we synthesized the thermally reactive polymer poly(N-isopropylacrylamide) (PNIPAM) using reversible addition–fragmentation chain transfer (RAFT) polymerization and prepared nanocarriers by binding PNIPAM to gold nanorods (GRs), with the anticancer agent doxorubicin (DOX) used as a model drug. PNIPAM changes from hydrophilic to hydrophobic at temperatures above its lower critical solution temperature, which represents a coil-to-globule volume phase transition. Because GRs absorb near-infrared (NIR) laser light and emit energy, PNIPAM aggregation occurs when the synthesized PNIPAM/GR are subjected to an NIR laser, and the temperature of the GRs rises. Using this principle, DOX was combined with the PNIPAM/GR complex, and the resulting anticancer effects with and without laser treatment were observed in Hela and MDA-MB-231 cells. In our proposed complex, the GR binding rate of PNIPAM reached 20% and the DOX binding rate reached 15%. The release profile of the drug following laser irradiation was determined using a drug release test and confocal microscopy imaging. It was subsequently confirmed that the release of the drug is higher at higher temperatures, especially with laser treatment. The proposed combination of temperature-reactive polymers and gold nanostructures shows promise for future research into controlled drug release.
A Solid-in-Oil Nanodispersion System for Transcutaneous Immunotherapy of Cow’s Milk Allergies
Authors: Kitaoka Momoko , Xiao Wei , Kong Qingliang , Tahara Yoshiro , Kamiya Noriho , Goto Masahiro
Published: March 2020

An allergy to cow’s milk proteins is the most common food allergy in infants and toddlers. Conventional oral immunotherapy for cow’s milk allergies requires hospital admission due to the risk of severe allergic reactions, including anaphylaxis. Therefore, a simpler and safer immunotherapeutic method is desirable. We examined transcutaneous immunotherapy with a solid-in-oil (S/O) system. In the S/O system, nano-sized particles of proteins are dispersed in an oil-vehicle with the assistance of nonionic surfactants. In the present study, the S/O system enhanced the skin permeation of the allergen molecule β-lactoglobulin (BLG), as compared with a control PBS solution. The patches containing BLG in the S/O nanodispersion skewed the immune response in the allergy model mice toward T helper type 1 immunity, indicating the amelioration of allergic symptoms. This effect was more pronounced when the immunomodulator resiquimod (R-848) was included in the S/O system.
Printability of External and Internal Structures Based on Digital Light Processing 3D Printing Technique
Authors: Yang Yan , Zhou Yanjun , Lin Xiao , Yang Qingliang , Yang Gengshen
Published: March 2020

The high printing efficiency and easy availability of desktop digital light processing (DLP) printers have made DLP 3D printing a promising technique with increasingly broad application prospects, particularly in personalized medicine. The objective of this study was to fabricate and evaluate medical samples with external and internal structures using the DLP technique. The influence of different additives and printing parameters on the printability and functionality of this technique was thoroughly evaluated. It was observed that the printability and mechanical properties of external structures were affected by the poly(ethylene glycol) diacrylate (PEGDA) concentration, plasticizers, layer height, and exposure time. The optimal printing solutions for 3D external and internal structures were 100% PEGDA and 75% PEGDA with 0.25 mg/mL tartrazine, respectively. And the optimal layer height for 3D external and internal structures were 0.02 mm and 0.05 mm, respectively. The optimal sample with external structures had an adequate drug-loading ability, acceptable sustained-release characteristics, and satisfactory biomechanical properties. In contrast, the printability of internal structures was affected by the photoabsorber, PEGDA concentration, layer height, and exposure time. The optimal samples with internal structures had good morphology, integrity and perfusion behavior. The present study showed that the DLP printing technique was capable of fabricating implants for drug delivery and physiological channels for in vivo evaluation.
Theoretical and Experimental Gas Volume Quantification of Micro- and Nanobubble Ultrasound Contrast Agents
Authors: Abenojar Eric C. , Bederman Ilya , de Leon Al C. , Zhu Jinle , Hadley Judith , Kolios Michael C. , Exner Agata A.
Published: March 2020

The amount of gas in ultrasound contrast agents is related to their acoustic activity. Because of this relationship, gas volume has been used as a key variable in normalizing the in vitro and in vivo acoustic behavior of lipid shell-stabilized bubbles with different sizes and shell components. Despite its importance, bubble gas volume has typically only been theoretically calculated based on bubble size and concentration that is typically measured using the Coulter counter for microbubbles and nanoparticle tracking analysis (NTA) for nanoscale bubbles. However, while these methods have been validated for the analysis of liquid or solid particles, their application in bubble analysis has not been rigorously studied. We have previously shown that resonant mass measurement (RMM) may be a better-suited technique for sub-micron bubble analysis, as it can measure both buoyant and non-buoyant particle size and concentration. Here, we provide validation of RMM bubble analysis by using headspace gas chromatography/mass spectrometry (GC/MS) to experimentally measure the gas volume of the bubble samples. This measurement was then used as ground truth to test the accuracy of theoretical gas volume predictions based on RMM, NTA (for nanobubbles), and Coulter counter (for microbubbles) measurements. The results show that the headspace GC/MS gas volume measurements agreed well with the theoretical predictions for the RMM of nanobubbles but not NTA. For nanobubbles, the theoretical gas volume using RMM was 10% lower than the experimental GC/MS measurements; meanwhile, using NTA resulted in an 82% lower predicted gas volume. For microbubbles, the experimental gas volume from the GC/MS measurements was 27% lower compared to RMM and 72% less compared to the Coulter counter results. This study demonstrates that the gas volume of nanobubbles and microbubbles can be reliably measured using headspace GC/MS to validate bubble size measurement techniques. We also conclude that the accuracy of theoretical predictions is highly dependent on proper size and concentration measurements.
Initiated Chemical Vapor Deposition of Crosslinked Organic Coatings for Controlling Gentamicin Delivery
Authors: Decandia Gianfranco , Palumbo Fabio , Treglia Annalisa , Armenise Vincenza , Favia Pietro , Baruzzi Federico , Unger Katrin , Perrotta Alberto , Coclite Anna Maria
Published: March 2020

A coating consisting of a copolymer of methacrylic acid and ethylene glycol dimethacrylate was deposited over a gentamicin film by initiated chemical vapor deposition with the aim of controlling the drug release. Gentamicin release in water was monitored by means of conductance measurements and of UV-vis Fluorescence Spectroscopy. The influence of the polymer chemical composition, specifically of its crosslinking density, has been investigated as a tool to control the swelling behavior of the initiated chemical vapor deposition (iCVD) coating in water, and therefore its ability to release the drug. Agar diffusion test and microbroth dilution assays against Staphylococcus aureus and Pseudomonas aeruginosa on cellulose coated substrates confirmed that the antibacterial activity of the drug released by the coating was retained, though the release of gentamicin was not complete.
Crystallisation Behaviour of Pharmaceutical Compounds Confined within Mesoporous Silicon
Authors: Jones Eleanor C. L. , Bimbo Luis M.
Published: March 2020

The poor aqueous solubility of new and existing drug compounds represents a significant challenge in pharmaceutical development, with numerous strategies currently being pursued to address this issue. Amorphous solids lack the repeating array of atoms in the structure and present greater free energy than their crystalline counterparts, which in turn enhances the solubility of the compound. The loading of drug compounds into porous materials has been described as a promising approach for the stabilisation of the amorphous state but is dependent on many factors, including pore size and surface chemistry of the substrate material. This review looks at the applications of mesoporous materials in the confinement of pharmaceutical compounds to increase their dissolution rate or modify their release and the influence of varying pore size to crystallise metastable polymorphs. We focus our attention on mesoporous silicon, due to the ability of its surface to be easily modified, enabling it to be stabilised and functionalised for the loading of various drug compounds. The use of neutron and synchrotron X-ray to examine compounds and the mesoporous materials in which they are confined is also discussed, moving away from the conventional analysis methods.
PGE2 Is Crucial for the Generation of FAST Whole- Tumor-Antigens Loaded Dendritic Cells Suitable for Immunotherapy in Glioblastoma
Authors: Nava Sara , Lisini Daniela , Frigerio Simona , Pogliani Simona , Pellegatta Serena , Gatti Laura , Finocchiaro Gaetano , Bersano Anna , Parati Eugenio Agostino
Published: March 2020

Dendritic cells (DC) are the most potent antigen-presenting cells, strongly inducers of T cell-mediated immune responses and, as such, broadly used as vaccine adjuvant in experimental clinical settings. DC are widely generated from human monocytes following in vitro protocols which require 5–7 days of differentiation with GM-CSF and IL-4 followed by 2–3 days of activation/maturation. In attempts to shorten the vaccine’s production, Fast-DC protocols have been developed. Here we reported a Fast-DC method in compliance with good manufacturing practices for the production of autologous mature dendritic cells loaded with antigens derived from whole tumor lysate, suitable for the immunotherapy in glioblastoma patients. The feasibility of generating Fast-DC pulsed with whole tumor lysate was assessed using a series of small-scale cultures performed in parallel with clinical grade large scale standard method preparations. Our results demonstrate that this Fast protocol is effective only in the presence of PGE2 in the maturation cocktail to guarantee that Fast-DC cells exhibit a mature phenotype and fulfill all requirements for in vivo use in immunotherapy approaches. Fast-DC generated following this protocol were equally potent to standard DC in inducing Ag-specific T cell proliferation in vitro. Generation of Fast-DC not only reduces labor, cost, and time required for in vitro clinical grade DC development, but can also minimizes inter-preparations variability and the risk of contamination.
Preparation of Puerarin Chitosan Oral Nanoparticles by Ionic Gelation Method and Its Related Kinetics
Authors: Yan Jie , Guan Zhi-Yu , Zhu Wei-Feng , Zhong Ling-Yun , Qiu Zhuo-Qi , Yue Peng-Fei , Wu Wen-Ting , Liu Jing , Huang Xiao
Published: March 2020

In this paper, as an active ingredient, puerarin chitosan nanoparticles (Pur-CS/TPP-NPs) are prepared by an ionic gelation method. The chitosan (CS) concentration, pH of the CS solution, sodium tripolyphosphate (TPP) concentration, stirring speed, stirring time, ultrasonic power, and dosage are used as single factors for investigation, and the encapsulation efficiency, drug loading capacity, particle size, and polydispersity index (PDI) are used as indicators for investigation. The optimal prescription is determined using the Box–Behnken effect surface design method. The characterization of the best formulation, which is determined via an in vitro release assay and liquid chromatography/tandem mass spectrometry (LC-MS/MS) analysis methods, is used here for pharmacokinetic studies. An in situ single-pass intestinal perfusion model is used to investigate drug absorption in the intestine. After characterization, the morphologies of the nanoparticles are intact. It can be seen from the in vitro release experiments that the equation fitted by the nanoparticles is the Higuchi model, the nanoparticle release process is very stable and without sudden release, indicating that the nanoparticles are well-released in vitro. The pharmacokinetic results and the in situ single-pass intestinal perfusion model study show that the bioavailability and absorption of Pur-CS/TPP-NPs were significantly higher than Pur. Thus, all the results show that the prepared nanoparticles can significantly improve the bioavailability of Pur, and we hope to lay the foundation for the development of new products of Pur.
Plant Nitrogen and Phosphorus Resorption in Response to Varied Legume Proportions in a Restored Grassland
Authors: Li Qiang , Zhou Daowei , Denton Matthew D.
Published: March 2020

An in-depth assessment of plant nutrient resorption can offer insights into understanding ecological processes and functional responses to biotic and abiotic changes in the environment. The legume proportion in a mixed grassland can drive changes in the soil environment and plant relationships, but little information is available regarding how the legume proportion influences plant nutrient resorption in mixed grasslands. In this study, three mixed communities of Leymus chinensis (Trin.) Tzvel. and Medicago sativa L. differing in legume proportion (Low-L, with 25% legume composition; Mid-L, with 50% legume composition; High-L, with 75% legume composition) were established with four replicates in a degraded grassland. Four years after establishing the mixed grassland, the quantity of biological N2 fixation by M. sativa, the availabilities of water and nitrogen (N) and phosphorus (P) in soil were examined, and the concentrations and resorption of leaf N and P for both species were measured during forage maturation and senescence. The results showed Mid-L had greater biological N2 fixation and soil N availability than Low-L and High-L, while the High-L had lower soil water and P availability, but a greater soil available N:P ratio compared with Low-L and Mid-L. Legume proportion did not alter N or P concentrations of mature leaves. However, in Mid-L N resorption was reduced by 8 to 16% for the two mixed-species compared with Low-L and High-L. High-L enhanced P resorption by 20 to 24% in both plant species compared with Low-L. The L. chinensis and M. sativa responded differently to varied legume proportion in terms of P resorption. It was concluded that legume proportion drove changes in soil nutrient availability of mixed communities, which primarily altered plant nutrient resorption during senescence, but had no influence on the nutrient concentrations of mature plants. A moderate legume proportion reduced N resorption, and increased senesced leaf N concentration of grass and legume species. The difference in P resorption by two mixed-species significantly changed the interspecific difference of senesced leaf P concentration and the N:P ratio with varied legume proportion.
Optimal Duration of Drought Stress Near Harvest for Promoting Bioactive Compounds and Antioxidant Capacity in Kale with or without UV-B Radiation in Plant Factories
Authors: Yoon Hyo In , Zhang Wenjuan , Son Jung Eek
Published: March 2020

Among abiotic stresses, both drought and UV-B radiation effectively trigger the accumulation of secondary metabolites, and can be widely applied in plant factories. The objectives of this study were to investigate antioxidant accumulation under drought stress alone, or in combination with UV-B radiation near harvest, and to determine an optimal treatment time for maximum antioxidant production. Kale (Brassica oleracea L. var. acephala) plants were grown in a plant factory and harvested at 42 days after transplanting. The single and combination treatments lasted for 7 to 1 days and 4 to 2 days before harvest, respectively. The results of both Fv/Fm (maximal photochemical efficiency in photosystem II) and leaf water potential could ensure the function of photosynthesis and maintain normal leaf moisture in single drought treatments of less than 4 days. The total phenolic and flavonoid contents and antioxidant activities were significantly increased in both single and combination treatments for 3 to 4 days, compared to other treatments. The supplementary UV-B treatments showed no extra formation of antioxidants compared to the single drought treatments. As a result, drought for 3 days before harvest could achieve the highest potential value of kale as a source of natural antioxidants.
Isatis tinctoria L. (Woad): A Review of Its Botany, Ethnobotanical Uses, Phytochemistry, Biological Activities, and Biotechnological Studies
Authors: Speranza Jasmine , Miceli Natalizia , Taviano Maria Fernanda , Ragusa Salvatore , Kwiecień Inga , Szopa Agnieszka , Ekiert Halina
Published: March 2020

Isatis tinctoria L. (Brassicaceae), which is commonly known as woad, is a species with an ancient and well-documented history as an indigo dye and medicinal plant. Currently, I. tinctoria is utilized more often as medicinal remedy and also as a cosmetic ingredient. In 2011, I. tinctoria root was accepted in the official European phytotherapy by introducing its monograph in the European Pharmacopoeia. The biological properties of raw material have been known from Traditional Chinese Medicine (TCM). Over recent decades, I. tinctoria has been investigated both from a phytochemical and a biological point of view. The modern in vitro and in vivo scientific studies proved anti-inflammatory, anti-tumour, antimicrobial, antiviral, analgesic, and antioxidant activities. The phytochemical composition of I. tinctoria has been thoroughly investigated and the plant was proven to contain many valuable biologically active compounds, including several alkaloids, among which tryptanthrin, indirubin, indolinone, phenolic compounds, and polysaccharides as well as glucosinolates, carotenoids, volatile constituents, and fatty acids. This article provides a general botanical and ethnobotanical overview that summarizes the up-to-date knowledge on the phytochemistry and biological properties of this valuable plant in order to support its therapeutic potential. Moreover, the biotechnological studies on I. tinctoria, which mainly focused on hairy root cultures for the enhanced production of flavonoids and alkaloids as well as on the establishment of shoot cultures and micropropagation protocols, were reviewed. They provide input for future research prospects.