bluuebunny/crossref_metadata_2025 huggingface dataset:
Exhaled Volatile Organic Compounds during Inflammation Induced by TNF-α in Ventilated Rats
Authors: Albrecht Frederic W. , Maurer Felix , Müller-Wirtz Lukas M. , Schwaiblmair Michaela H. , Hüppe Tobias , Wolf Beate , Sessler Daniel I. , Volk Thomas , Kreuer Sascha , Fink Tobias
Published: June 2020

Systemic inflammation alters the composition of exhaled breath, possibly helping clinicians diagnose conditions such as sepsis. We therefore evaluated changes in exhaled breath of rats given tumor necrosis factor-alpha (TNF-α). Thirty male Sprague-Dawley rats were randomly assigned to three groups (n = 10 each) with intravenous injections of normal saline (control), 200 µg·kg−1 bodyweight TNF-α (TNF-α-200), or 600 µg·kg−1 bodyweight TNF-α (TNF-α-600), and were observed for 24 h or until death. Animals were ventilated with highly-purified synthetic air to analyze exhaled air by multicapillary column–ion mobility spectrometry. Volatile organic compounds (VOCs) were identified from a database. We recorded blood pressure and cardiac output, along with cytokine plasma concentrations. Control rats survived the 24 h observation period, whereas mean survival time decreased to 22 h for TNF-α-200 and 23 h for TNF-α-600 rats. Mean arterial pressure decreased in TNF-α groups, whereas IL-6 increased, consistent with mild to moderate inflammation. Hundreds of VOCs were detected in exhalome. P-cymol increased by a factor-of-two 4 h after injection of TNF-α-600 compared to the control and TNF-α-200. We found that 1-butanol and 1-pentanol increased in both TNF-α groups after 20 h compared to the control. As breath analysis distinguishes between two doses of TNF-α and none, we conclude that it might help clinicians identify systemic inflammation.
Pursuing Experimental Reproducibility: An Efficient Protocol for the Preparation of Cerebrospinal Fluid Samples for NMR-Based Metabolomics and Analysis of Sample Degradation
Authors: Albrecht Benjamin , Voronina Elena , Schipke Carola , Peters Oliver , Parr Maria Kristina , Díaz-Hernández M. Dolores , Schlörer Nils E.
Published: June 2020

NMR-based metabolomics investigations of human biofluids offer great potential to uncover new biomarkers. In contrast to protocols for sample collection and biobanking, procedures for sample preparation prior to NMR measurements are still heterogeneous, thus compromising the comparability of the resulting data. Herein, we present results of an investigation of the handling of cerebrospinal fluid (CSF) samples for NMR metabolomics research. Origins of commonly observed problems when conducting NMR experiments on this type of sample are addressed, and suitable experimental conditions in terms of sample preparation and pH control are discussed. Sample stability was assessed by monitoring the degradation of CSF samples by NMR, hereby identifying metabolite candidates, which are potentially affected by sample storage. A protocol was devised yielding consistent spectroscopic data as well as achieving overall sample stability for robust analysis. We present easy to adopt standard operating procedures with the aim to establish a shared sample handling strategy that facilitates and promotes inter-laboratory comparison, and the analysis of sample degradation provides new insights into sample stability.
First Detection of Paralytic Shellfish Toxins from Alexandrium pacificum above the Regulatory Limit in Blue Mussels (Mytilus galloprovincialis) in New South Wales, Australia
Authors: Barua Abanti , Ajani Penelope A. , Ruvindy Rendy , Farrell Hazel , Zammit Anthony , Brett Steve , Hill David , Sarowar Chowdhury , Hoppenrath Mona , Murray Shauna A.
Published: June 2020

In 2016, 2017 and 2018, elevated levels of the species Alexandrium pacificum were detected within a blue mussel (Mytilus galloprovincialis) aquaculture area at Twofold Bay on the south coast of New South Wales, Australia. In 2016, the bloom persisted for at least eight weeks and maximum cell concentrations of 89,000 cells L−1 of A. pacificum were reported. The identity of A. pacificum was confirmed using molecular genetic tools (qPCR and amplicon sequencing) and complemented by light and scanning electron microscopy of cultured strains. Maximum reported concentrations of paralytic shellfish toxins (PSTs) in mussel tissue was 7.2 mg/kg PST STX equivalent. Elevated cell concentrations of A. pacificum were reported along the adjacent coastal shelf areas, and positive PST results were reported from nearby oyster producing estuaries during 2016. This is the first record of PSTs above the regulatory limit (0.8 mg/kg) in commercial aquaculture in New South Wales since the establishment of routine biotoxin monitoring in 2005. The intensity and duration of the 2016 A. pacificum bloom were unusual given the relatively low abundances of A. pacificum in estuarine and coastal waters of the region found in the prior 10 years.
Meta-Analysis of Microbial Communities in Hot Springs: Recurrent Taxa and Complex Shaping Factors beyond pH and Temperature
Authors: Massello Francisco L. , Chan Chia Sing , Chan Kok-Gan , Goh Kian Mau , Donati Edgardo , Urbieta María Sofía
Published: June 2020

The study of microbial communities from extreme environments is a fascinating topic. With every study, biologists and ecologists reveal interesting facts and questions that dispel the old belief that these are inhospitable environments. In this work, we assess the microbial diversity of three hot springs from Neuquén, Argentina, using high-throughput amplicon sequencing. We predicted a distinct metabolic profile in the acidic and the circumneutral samples, with the first ones being dominated by chemolithotrophs and the second ones by chemoheterotrophs. Then, we collected data of the microbial communities of hot springs around the world in an effort to comprehend the roles of pH and temperature as shaping factors. Interestingly, there was a covariation between both parameters and the phylogenetic distance between communities; however, neither of them could explain much of the microbial profile in an ordination model. Moreover, there was no correlation between alpha diversity and these parameters. Therefore, the microbial communities’ profile seemed to have complex shaping factors beyond pH and temperature. Lastly, we looked for taxa associated with different environmental conditions. Several such taxa were found. For example, Hydrogenobaculum was frequently present in acidic springs, as was the Sulfolobaceae family; on the other hand, Candidatus Hydrothermae phylum was strongly associated with circumneutral conditions. Interestingly, some singularities related to sites featuring certain taxa were also observed.
Interrupted Blood Feeding in Ticks: Causes and Consequences
Authors: Tahir Djamel , Meyer Leon , Fourie Josephus , Jongejan Frans , Mather Thomas , Choumet Valérie , Blagburn Byron , Straubinger Reinhard K. , Varloud Marie
Published: June 2020

Ticks are obligate hematophagous arthropods and act as vectors for a great variety of pathogens, including viruses, bacteria, protozoa, and helminths. Some tick-borne viruses, such as Powassan virus and tick-borne encephalitis virus, are transmissible within 15–60 min after tick attachment. However, a minimum of 3–24 h of tick attachment is necessary to effectively transmit bacterial agents such as Ehrlichia spp., Anaplasma spp., and Rickettsia spp. to a new host. Longer transmission periods were reported for Borrelia spp. and protozoans such as Babesia spp., which require a minimum duration of 24–48 h of tick attachment for maturation and migration of the pathogen. Laboratory observations indicate that the probability of transmission of tick-borne pathogens increases with the duration an infected tick is allowed to remain attached to the host. However, the transmission time may be shortened when partially fed infected ticks detach from their initial host and reattach to a new host, on which they complete their engorgement. For example, early transmission of tick-borne pathogens (e.g., Rickettsia rickettsii, Borrelia burgdorferi, and Babesia canis) and a significantly shorter transmission time were demonstrated in laboratory experiments by interrupted blood feeding. The relevance of such situations under field conditions remains poorly documented. In this review, we explore parameters of, and causes leading to, spontaneous interrupted feeding in nature, as well as the effects of this behavior on the minimum time required for transmission of tick-borne pathogens.
A Municipality-Based Approach Using Commuting Census Data to Characterize the Vulnerability to Influenza-Like Epidemic: The COVID-19 Application in Italy
Authors: Savini Lara , Candeloro Luca , Calistri Paolo , Conte Annamaria
Published: June 2020

In February 2020, Italy became the epicenter for COVID-19 in Europe, and at the beginning of March, the Italian Government put in place emergency measures to restrict population movement. Aim of our analysis is to provide a better understanding of the epidemiological context of COVID-19 in Italy, using commuting data at a high spatial resolution, characterizing the territory in terms of vulnerability. We used a Susceptible–Infectious stochastic model and we estimated a municipality-specific infection contact rate (β) to capture the susceptibility to the disease. We identified in Lombardy, Veneto and Emilia Romagna regions (52% of all Italian cases) significant clusters of high β, due to the simultaneous presence of connections between municipalities and high population density. Local simulated spreading in regions, with different levels of infection observed, showed different disease geographical patterns due to different β values and commuting systems. In addition, we produced a vulnerability map (in the Abruzzi region as an example) by simulating the epidemic considering each municipality as a seed. The result shows the highest vulnerability values in areas with commercial hubs, close to the highest populated cities and the most industrial area. Our results highlight how human mobility can affect the epidemic, identifying particular situations in which the health authorities can promptly intervene to control the disease spread.
Mineralogy and Geochemistry of Sr-Bearing Phosphates from the Nanping No. 31 Pegmatite (SE China): Implications for Sr Circulation and Post-Magmatic Processes in Granitic Systems
Authors: Rao Can , Wang Ru-Cheng , Hatert Frédéric , Wu Run-Qiu , Wang Qi
Published: June 2020

Although Rb/Sr ratio and Sr isotopes are routinely modeled to determine petrogenetic processes and sources for granitic systems, the post-magmatic path and crystallization of Sr in granitic systems have not been thoroughly elucidated to date. In this study, we present the petrography, chemical composition and isotopic 87Sr/86 of Sr-bearing phosphates from the Nanping No. 31 pegmatite in Southeastern China, helping to characterize post-magmatic stages and geochemical recirculation of Sr in granitic systems. K-feldspar and primary apatites occur as major “primary Sr minerals”, the occurrences of secondary Sr phosphates (strontiohurlbutite, palermoite and goyazite) and Sr-rich phosphates (apatites, hurlbutite, bertossaite and fluorarrojadite-(BaNa)) reflect the transport, concentration and recrystallization of Sr in granitic systems. The mobilization and recrystallization of Sr in granitic systems are mainly controlled by the variation in alkalinity of hydrothermal fluids. Two post-magmatic recirculations of Sr are proposed in the Nanping No. 31 pegmatite: (1) breakdown of the “primary Sr mineral” (K-feldspar and primary apatites) and crystallization of secondary Sr-bearing phases; and (2) replacement of secondary Sr-bearing phosphates and direct precipitation of later palermoite and goyazite from later Sr-rich fluids at low temperatures. The Sr isotope features of Sr-bearing phosphates suggest that the emplacement and consolidation processes of the Nanping pegmatite involved the participation of externally derived fluids.
Origin of a Petrographic Coal Structure and Its Implication for Coalbed Methane Evaluation
Authors: Cheng Wei , Yang Ruidong , Zhang Qin
Published: June 2020

A petrographic coal structure of Late Permian coals from the Liupanshui coalfield, Western Guizhou, SW China, has been distinguished for its novel macro-lithological characteristics. Petrographic, mineralogical and geochemical studies have been conducted for a typical coal sample (No.3 coal, Songhe coalmine, Panzhou County, China) and its geological genesis and significance for coalbed methane (CBM) evaluation is accordingly discussed. It was found that coal is characterized by a banded structure with intensively fractured vitrain sublayers, where a great number of fractures were developed and filled with massive inorganic matter. The study of coal quality, coal petrography, mineralogy and lanthanides and yttrium (REY) geochemistry of the infilling mineral matter (IMM) indicates that this fractured coal structure resulted from the tissues of coal-forming plants or coal matrix shrinkage, as well as the precipitation of calcium rich groundwater and the addition of terrigenous materials. The coal depositional environment and coal-forming plant are considered to have played a role in inducing the special fractures. This provides a scientific reference for the study of CBM for coal with this fractured structure, such as the Late Permian coal from the western border of Guizhou Province, SW China.
Effect of the Fluorination of Graphene Nanoflake on the Dispersion and Mechanical Properties of Polypropylene Nanocomposites
Authors: Lee Min Gyu , Lee Sangwoon , Cho Jaehyun , Bae Seokyoung , Jho Jae Young
Published: June 2020

In order to investigate the effect of fluorination of graphene nanoflake on the dispersibility in polypropylene (PP) composites, fluorinated graphene oxide (FGO) was prepared by solvo-thermal reaction and applied as a filler of the PP nanocomposite. Due to the weakened inter-particle attraction among the graphene nanoflake and reduced surface energy difference between PP and the filler, PP/FGO composites showed better exfoliation and dispersion state of the filler compared with that of PP/graphene oxide (GO) or PP/reduced graphene oxide (RGO) composites. The improved exfoliation and dispersion of graphene nanoflake resulted in a significant reinforcement on the composites. The Young’s modulus and tensile strength of PP composites filled with 2 wt% of FGO increased by 31% and 15%, respectively, compared with those of PP.
Biotin-Conjugated Cellulose Nanofibers Prepared via Copper-Catalyzed Alkyne-Azide Cycloaddition (CuAAC) “Click” Chemistry
Authors: Goodge Katarina , Frey Margaret
Published: June 2020

As potential high surface area for selective capture in diagnostic or filtration devices, biotin-cellulose nanofiber membranes were fabricated to demonstrate the potential for specific and bio-orthogonal attachment of biomolecules onto nanofiber surfaces. Cellulose acetate was electrospun and substituted with alkyne groups in either a one- or two-step process. The alkyne reaction, confirmed by FTIR and Raman spectroscopy, was dependent on solvent ratio, time, and temperature. The two-step process maximized alkyne substitution in 10/90 volume per volume ratio (v/v) water to isopropanol at 50 °C after 6 h compared to the one-step process in 80/20 (v/v) at 50 °C after 48 h. Azide-biotin conjugate “clicked” with the alkyne-cellulose via copper-catalyzed alkyne-azide cycloaddition (CuAAC). The biotin-cellulose membranes, characterized by FTIR, SEM, Energy Dispersive X-ray spectroscopy (EDX), and XPS, were used in proof-of-concept assays (HABA (4′-hydroxyazobenzene-2-carboxylic acid) colorimetric assay and fluorescently tagged streptavidin assay) where streptavidin selectively bound to the pendant biotin. The click reaction was specific to alkyne-azide coupling and dependent on pH, ratio of ascorbic acid to copper sulfate, and time. Copper (II) reduction to copper (I) was successful without ascorbic acid, increasing the viability of the click conjugation with biomolecules. The surface-available biotin was dependent on storage medium and time: Decreasing with immersion in water and increasing with storage in air.
Sonication-Assisted Production of Fosetyl-Al Nanocrystals: Investigation of Human Toxicity and In Vitro Antibacterial Efficacy against Xylella fastidiosa
Authors: Baldassarre Francesca , Tatulli Giuseppe , Vergaro Viviana , Mariano Stefania , Scala Valeria , Nobile Concetta , Pucci Nicoletta , Dini Luciana , Loreti Stefania , Ciccarella Giuseppe
Published: June 2020

Recently, there is a growing demand in sustainable phytopathogens control research. Nanotechnology provides several tools such as new pesticides formulations, antibacterial nanomaterials and smart delivery systems. Metal nano-oxides and different biopolymers have been exploited in order to develop nanopesticides which can offer a targeted solution minimizing side effects on environment and human health. This work proposed a nanotechnological approach to obtain a new formulation of systemic fungicide fosetyl-Al employing ultrasonication assisted production of water dispersible nanocrystals. Moreover, chitosan was applicated as a coating agent aiming a synergistic antimicrobial effect between biopolymer and fungicide. Fosetyl-Al nanocrystals have been characterized by morphological and physical-chemical analysis. Nanotoxicological investigation was carried out on human keratinocytes cells through cells viability test and ultrastructural analysis. In vitro planktonic growth, biofilm production and agar dilution assays have been conducted on two Xylella fastidiosa subspecies. Fosetyl-Al nanocrystals resulted very stable over time and less toxic respect to conventional formulation. Finally, chitosan-based fosetyl-Al nanocrystals showed an interesting antibacterial activity against Xylella fastidiosa subsp. pauca and Xylella fastidiosa subsp. fastidiosa.
pH-Responsive Resveratrol-Loaded Electrospun Membranes for the Prevention of Implant-Associated Infections
Authors: Bonadies Irene , Di Cristo Francesca , Valentino Anna , Peluso Gianfranco , Calarco Anna , Di Salle Anna
Published: June 2020

To date, the implant-associated infections represent a worldwide challenge for the recently reported bacterial drug resistance that can lead to the inefficacy or low efficacy of conventional antibiotic therapies. Plant polyphenolic compounds, including resveratrol (RSV), are increasingly gaining consensus as valid and effective alternatives to antibiotics limiting antibiotic resistance. In this study, electrospun polylactic acid (PLA) membranes loaded with different concentrations of RSV are synthesized and characterized in their chemical, morphological, and release features. The obtained data show that the RSV release rate from the PLA-membranes is remarkably higher in acidic conditions than at neutral pH. In addition, a change in pH from neutral to slightly acidic triggers a significant increase in the RSV release. This behavior indicates that the PLA-RSV membranes can act as drug reservoir when the environmental pH is neutral, starting to release the bioactive molecules when the pH decreases, as in presence of oral bacterial infection. Indeed, our results demonstrate that PLA-RSV2 displays a significant antibacterial and antibiofilm activity against two bacterial strains, Pseudomonas aeruginosa PAO1, and Streptococcus mutans, responsible for both acute and chronic infections in humans, thus representing a promising solution for the prevention of the implant-associated infections.
SNP rs6564851 in the BCO1 Gene Is Associated with Varying Provitamin a Plasma Concentrations but Not with Retinol Concentrations among Adolescents from Rural Ghana
Authors: Graßmann Sophie , Pivovarova-Ramich Olga , Henze Andrea , Raila Jens , Ampem Amoako Yaw , King Nyamekye Richard , Bedu-Addo George , Mockenhaupt Frank , Schulze Matthias , Danquah Ina
Published: June 2020

In sub-Saharan Africa, vitamin A deficiency constitutes a severe health problem despite various supplementation and food fortification programs. Given that the intake of preformed vitamin A from animal products remains low in these countries, an efficient metabolization of plant-based provitamin A carotenoids is essential. Previously, adolescents in rural Ghana have shown high total plasma carotenoid concentrations, while 36% had a vitamin A deficiency (defined as plasma retinol < 0.7 µmol/L). Hence, the aim of this cross-sectional study was to identify the relationships between variants in the β-carotene 15,15’-oxygenase (BCO1) gene and plasma carotenoid concentrations among 189 15-year-old girls and boys in rural Ghana. BCO1 rs6564851, rs7500996, rs10048138 and PKD1L2 rs6420424, and rs8044334 were typed, and carotenoid concentrations were compared among the different genotypes. G allele carriers of rs6564851 (53%) showed higher plasma carotenoid concentrations than T allele carriers (median (interquartile range): 3.07 (2.17–4.02) vs. 2.59 (2.21–3.50) µmol/L, p-value = 0.0424). This was not explained by differences in socio-demographic or dietary factors. In contrast, no differences in plasma retinol concentrations were observed between these genotypes. Pending verification in independent populations, the low conversion efficiency of provitamin A carotenoids among rs6564851 G allele carriers may undermine existing fortification and supplementation programs to improve the vitamin A status in sub-Saharan Africa.
Usual Dietary Intake, Nutritional Adequacy and Food Sources of Calcium, Phosphorus, Magnesium and Vitamin D of Spanish Children Aged One to <10 Years. Findings from the EsNuPI Study
Authors: Cuadrado-Soto Esther , López-Sobaler Ana M. , Jiménez-Ortega Ana Isabel , Aparicio Aránzazu , Bermejo Laura M. , Hernández-Ruiz Ángela , Lara Villoslada Federico , Leis Rosaura , Martínez de Victoria Emilio , Moreno José Manuel , Ruiz-López María Dolores , Soto-Méndez María José , Valero Teresa , Varela-Moreiras Gregorio , Gil Ángel , Ortega Rosa M.
Published: June 2020

Bone problems in the population begin to be establish in childhood. The present study aims to assess the usual calcium, phosphorus, magnesium, and vitamin D intakes, along with the food sources of these nutrients, in Spanish children participating in the EsNuPI (Estudio Nutricional en Población Infantil Española) study. Two 24 h dietary recalls were applied to 1448 children (1 to <10 years) divided into two sub-samples: one reference sample (RS) of the general population [n = 707] and another sample which exclusively included children consuming enriched or fortified milks, here called “adapted milks” (AMS) [n = 741]. Estimation of the usual intake shows that nutrient intake increased with age for all nutrients except vitamin D. Using as reference the Dietary Reference Values from the European Food Safety Authority (EFSA), calcium and magnesium intakes were found to be below the average requirement (AR) and adequate intake (AI), respectively, in a considerable percentage of children. Furthermore, phosphorus exceeded the AI in 100% of individuals and vitamin D was lower than the AI in almost all children studied. The results were very similar when considering only plausible reporters. When analyzing the food sources of the nutrients studied, milk and dairy products contributed the most to calcium, phosphorus, magnesium, and vitamin D. Other sources of calcium were cereals and vegetables; for phosphorus: meat, meat products, and cereals; for magnesium: cereals and fruits; and, for vitamin D: fish and eggs. These results highlight the desirability of improving the intake concerning these nutrients, which are involved in bone and metabolic health in children. The AMS group appeared to contribute better to the adequacy of those nutrients than the RS group, but both still need further improvement. Of special interest are the results of vitamin D intakes, which were significantly higher in the AMS group (although still below the AI), independent of age.
Identification of lncRNAs Involved in PCV2 Infection of PK-15 Cells
Authors: He Jin , Leng Chaoliang , Pan Jiazhen , Li Aoqi , Zhang Hua , Cong Feng , Wang Huanan
Published: June 2020

Porcine circovirus type 2 (PCV2) can cause severe disease in infected pigs, resulting in massive economic loss for the swine industry. Transcriptomic and proteomic approaches have been widely employed to identify the underlying molecular mechanisms of the PCV2 infection. Numerous differentially expressed mRNAs, miRNAs, and proteins, together with their associated signaling pathways, have been identified during PCV2 infection, paving the way for analysis of their biological functions. Long noncoding RNAs (lncRNAs) are important regulators of multiple biological processes. However, little is known regarding their role in the PCV2 infection. Hence, in our study, RNA-seq was performed by infecting PK-15 cells with PCV2. Analysis of the differentially expressed genes (DEGs) suggested that the cytoskeleton, apoptosis, cell division, and protein phosphorylation were significantly disturbed. Then, using stringent parameters, six lncRNAs were identified. Additionally, potential targets of the lncRNAs were predicted using both cis- and trans-prediction methods. Interestingly, we found that the HOXB (Homeobox B) gene cluster was probably the target of the lncRNA LOC106505099. Enrichment analysis of the target genes showed that numerous developmental processes were altered during PCV2 infection. Therefore, our study revealed that lncRNAs might affect porcine embryonic development through the regulation of the HOXB genes.
Development of an Innovative, Carrier-Based Dry Powder Inhalation Formulation Containing Spray-Dried Meloxicam Potassium to Improve the In Vitro and In Silico Aerodynamic Properties
Authors: Benke Edit , Farkas Árpád , Szabó-Révész Piroska , Ambrus Rita
Published: June 2020

Most of the marketed dry powder inhalation (DPI) products are traditional, carrier-based formulations with low drug concentrations deposited in the lung. However, due to their advantageous properties, their development has become justified. In our present work, we developed an innovative, carrier-based DPI system, which is an interactive physical blend of a surface-modified carrier and a spray-dried drug with suitable shape and size for pulmonary application. Meloxicam potassium, a nonsteroidal anti-inflammatory drug (NSAID), was used as an active ingredient due to its local anti-inflammatory effect and ability to decrease the progression of cystic fibrosis (CF) and chronic obstructive pulmonary disease (COPD). The results of the in vitro and in silico investigations showed high lung deposition in the case of this new formulation, confirming that the interparticle interactions were changed favorably.
pH-Sensitive Folic Acid Conjugated Alginate Nanoparticle for Induction of Cancer-Specific Fluorescence Imaging
Authors: Lee Sara , Lee Kangwon
Published: June 2020

In cancer nanomedicine, numerous studies have been conducted on the surface modification and transport capacity of nanoparticles (NPs); however, biological barriers, such as enzymatic degradation or non-specific delivery during circulation, remain to be cleared. Herein, we developed pH-sensitive NPs that degrade in an acidic environment and release 5-aminolevulinic acid (5ALA) to the target site. NPs were prepared by conjugating alginate with folic acid, followed by encapsulation of 5ALA through a water-in-oil (W/O) emulsion method. The alginate-conjugated folic acid nanoparticles (AF NPs) were homogeneous in size, stable for a long time in aqueous suspension without aggregation, and non-toxic. AF NPs were small enough to efficiently infiltrate tumors (<50 nm) and were specifically internalized by cancer cells through receptor-mediated endocytosis. After the intracellular absorption of NPs, alginate was deprotonated in the lysosomes and released 5ALA, which was converted to protoporphyrin IX (PpIX) through mitochondrial heme synthesis. Our study outcomes demonstrated that AF NPs were not degraded by enzymes or other external factors before reaching cancer cells, and fluorescent precursors were specifically and accurately delivered to cancer cells to generate fluorescence.
A Novel Composite Hydrogel Composed of Formic Acid-Decellularized Pepsin-Soluble Extracellular Matrix Hydrogel and Sacchachitin Hydrogel as Wound Dressing to Synergistically Accelerate Diabetic Wound Healing
Authors: Hsieh Chien-Ming , Wang Weu , Chen Ying-Hsuan , Wei Pu-Sheng , Liu Yu-Hsuan , Sheu Ming-Thau , Ho Hsiu-O
Published: June 2020

Extracellular matrix (ECM) hydrogel can create a favorable regenerative microenvironment and act as a promising dressing for accelerating the healing of diabetic wound. In this study, a simple and effective decellularization technique was developed and optimized to obtain acellular extracellular matrix (aECM) from porcine skin. It was found that decellularization at 30% formic acid for 72 h effectively decellularized porcine skin while retaining >75% collagen and ~37% GAG in the aECM with no presence of nuclei of cellular remnants. aECM hydrogel was fabricated by digesting aECM with pepsin in various acidic solutions (0.1 N HCl, glycolic acid (GA) and 2-pyrrolidone-5-carboxylic acid (PCA)) and then treated with a pH-controlled neutralization and temperature-controlled gelation procedure. Based on physical characterizations, including SDS-PAGE, rheological analysis and SEM analysis, aECMHCl hydrogels fabricated at 25 mg/mL in 0.1 N HCl were selected. Four polymeric ECM-mimic hydrogels, including sacchachitin (SC), hyaluronic acid (HA) and chitosan (CS) and three composite hydrogels of combining SC either with aECMHCl,25 (aECMHCl/SC), HA (HA/SC) or CS (SC/CS) were prepared and evaluated for WS-1 cell viability and wound-healing effectiveness. Cell viability study confirmed that no hydrogel dressings possessed any toxicity at all concentrations examined and ECMHCl, HA and ECMHCl/SC at higher concentrations (>0.05%) induced statistically significant proliferation. Diabetic wound healing study and histological examinations revealed that ECMHCl/SC hydrogel was observed to synergistically accelerate wound healing and ultimately stimulated the growth of hair follicles and sweat glands in the healing wound indicating the wound had healed as functional tissues. The results support the great potential of this newly produced ECMHCl/SC composite hydrogel for healing and regeneration of diabetic wounds.
Review: Local Tumor Necrosis Factor-α Inhibition in Inflammatory Bowel Disease
Authors: Gareb Bahez , Otten Antonius T. , Frijlink Henderik W. , Dijkstra Gerard , Kosterink Jos G. W.
Published: June 2020

Crohn’s disease (CD) and ulcerative colitis (UC) are inflammatory bowel diseases (IBD) characterized by intestinal inflammation. Increased intestinal levels of the proinflammatory cytokine tumor necrosis factor-α (TNF-α) are associated with disease activity and severity. Anti-TNF-α therapy is administered systemically and efficacious in the treatment of IBD. However, systemic exposure is associated with adverse events that may impede therapeutic treatment. Clinical studies show that the efficacy correlates with immunological effects localized in the gastrointestinal tract (GIT) as opposed to systemic effects. These data suggest that site-specific TNF-α inhibition in IBD may be efficacious with fewer expected side effects related to systemic exposure. We therefore reviewed the available literature that investigated the efficacy or feasibility of local TNF-α inhibition in IBD. A literature search was performed on PubMed with given search terms and strategy. Of 8739 hits, 48 citations were included in this review. These studies ranged from animal studies to randomized placebo-controlled clinical trials. In these studies, local anti-TNF-α therapy was achieved with antibodies, antisense oligonucleotides (ASO), small interfering RNA (siRNA), microRNA (miRNA) and genetically modified organisms. This narrative review summarizes and discusses these approaches in view of the clinical relevance of local TNF-α inhibition in IBD.
Development and Evaluation of Docetaxel-Phospholipid Complex Loaded Self-Microemulsifying Drug Delivery System: Optimization and In Vitro/Ex Vivo Studies
Authors: Wang Miao , You Sung-Kyun , Lee Hong-Ki , Han Min-Gu , Lee Hyeon-Min , Pham Thi Mai Anh , Na Young-Guk , Cho Cheong-Weon
Published: June 2020

Docetaxel (DTX) has clinical efficacy in the treatment of breast cancer, but it is difficult to develop a product for oral administration, due to low solubility and permeability. This study focused on preparing a self-microemulsifying drug delivery system (SME) loaded with DTX-phospholipid complex (DTX@PLC), to improve the dissolution and gastrointestinal (GI) permeability of DTX. A dual technique combining the phospholipid complexation and SME formulation described as improving upon the disadvantages of DTX has been proposed. We hypothesized that the complexation of DTX with phospholipids can improve the lipophilicity of DTX, thereby increasing the affinity of the drug to the cell lipid membrane, and simultaneously improving permeability through the GI barrier. Meanwhile, DTX@PLC-loaded SME (DTX@PLC-SME) increases the dissolution and surface area of DTX by forming a microemulsion in the intestinal fluid, providing sufficient opportunity for the drug to contact the GI membrane. First, we prepared DTX@PLC-SME by combining dual technologies, which are advantages for oral absorption. Next, we optimized DTX@PLC-SME with nanosized droplets (117.1 nm), low precipitation (8.9%), and high solubility (33.0 mg/g), which formed a homogeneous microemulsion in the aqueous phase. Dissolution and cellular uptake studies demonstrated that DTX@PLC-SME showed 5.6-fold higher dissolution and 2.3-fold higher DTX uptake in Caco-2 cells than raw material. In addition, an ex vivo gut sac study confirmed that DTX@PLC-SME improved GI permeability of DTX by 2.6-fold compared to raw material. These results suggested that DTX@PLC-SME can significantly overcome the disadvantages of anticancer agents, such as low solubility and permeability.
Preclinical Development of Autologous Hematopoietic Stem Cell-Based Gene Therapy for Immune Deficiencies: A Journey from Mouse Cage to Bed Side
Authors: Garcia-Perez Laura , Ordas Anita , Canté-Barrett Kirsten , Meij Pauline , Pike-Overzet Karin , Lankester Arjan , Staal Frank J. T.
Published: June 2020

Recent clinical trials using patient’s own corrected hematopoietic stem cells (HSCs), such as for primary immunodeficiencies (Adenosine deaminase (ADA) deficiency, X-linked Severe Combined Immunodeficiency (SCID), X-linked chronic granulomatous disease (CGD), Wiskott–Aldrich Syndrome (WAS)), have yielded promising results in the clinic; endorsing gene therapy to become standard therapy for a number of diseases. However, the journey to achieve such a successful therapy is not easy, and several challenges have to be overcome. In this review, we will address several different challenges in the development of gene therapy for immune deficiencies using our own experience with Recombinase-activating gene 1 (RAG1) SCID as an example. We will discuss product development (targeting of the therapeutic cells and choice of a suitable vector and delivery method), the proof-of-concept (in vitro and in vivo efficacy, toxicology, and safety), and the final release steps to the clinic (scaling up, good manufacturing practice (GMP) procedures/protocols and regulatory hurdles).
Bioprinting and Preliminary Testing of Highly Reproducible Novel Bioink for Potential Skin Regeneration
Authors: Hafezi Forough , Shorter Susan , Tabriz Atabak Ghanizadeh , Hurt Andrew , Elmes Victoria , Boateng Joshua , Douroumis Dennis
Published: June 2020

Three-dimensional (3D) bioprinting is considered as a novel approach in biofabricating cell-laden constructs that could potentially be used to promote skin regeneration following injury. In this study, a novel crosslinked chitosan (CH)–genipin (GE) bioink laden with keratinocyte and human dermal fibroblast cells was developed and printed successfully using an extruder-based bioprinter. By altering the composition and degree of CH–GE crosslinking, bioink printability was further assessed and compared with a commercial bioink. Rheological analysis showed that the viscosity of the optimised bioink was in a suitable range that facilitated reproducible and reliable printing by applying low pressures ranging from 20–40 kPa. The application of low printing pressures proved vital for viability of cells loaded within the bioinks. Further characterisation using MTT assay showed that cells were still viable within the printed construct at 93% despite the crosslinking, processing and after subjecting to physiological conditions for seven days. The morphological study of the printed cells showed that they were mobile within the bioink. Furthermore, the multi-layered 3D printed constructs demonstrated excellent self-supportive structures in a consistent manner.
Shear Stress-Dependent Targeting Efficiency Using Self-Assembled Gelatin–Oleic Nanoparticles in a Biomimetic Microfluidic System
Authors: Kang Taehee , Park Chulhun , Meghani Nileshkumar , Tran Thao T.D. , Tran Phuong H.L. , Lee Beom-Jin
Published: June 2020

Cellular properties and microenvironments, as well as the characteristics of nanoparticles (NPs), affect the cellular uptake and cytotoxic effects of drug-loaded NPs. Since there is fluid flow in the human blood system, fluid flow also affects the drug delivery efficiency of NPs. This study aimed to evaluate the cellular behaviors of drug-loaded soft NPs on A549 cancer cells under different levels of shear stress (0.5, 5, and 50 dynes/cm2) in the biomimetic microfluidic system. The soft self-assembled NPs were formed by the gelatin–oleic conjugate (GOC). The poorly water-soluble coumarin-6 or paclitaxel (PTX) were used as model markers for encapsulation within self-assembled NPs (C-GONs or PTX-GONs, respectively). The cellular uptake of C-GONs was found to be improved with shear-stress dependence. The inhibitory concentration (IC50) of PTX-GONs at 0.5, 5, and 50 dynes/cm2 was 0.106 µg/mL, 0.108 µg/mL, and 0.091 µg/mL, respectively, as compared to 0.138 µg/mL in a static condition. The cell killing efficiency of PTX-GONs was increased in the highest shear stress of 50 dynes/cm2 in the static condition, and other levels of shear stress in dynamic conditions.
Light Interception and the Growth of Pastures under Ideal and Stressful Growing Conditions on the Allegheny Plateau
Authors: Rayburn Edward B. , Griggs Thomas C.
Published: June 2020

Pasture-based livestock production is impacted by management and weather. In pastures, there is conflict between leaf retention for plant growth and leaf harvest for animal nutrition. Defoliated pastures with low light interception (LI) may have a low forage growth rate (FGR), while excessive growth shades leaves, reducing FGR and resulting in an S-shaped regrowth curve. To optimize production, it is best to keep FGR linear. Three studies were conducted to evaluate the impact of management and weather on FGR. Replicated pastures were used to measure FGR when grazed from 25 to 10 cm and allowed to regrow. The impact of alternative defoliation timings and intensities on FGR were studied using clipped treatments at three recovery intervals and two stubble heights. Variability in FGR was studied using a field validated plant growth model. Of the 24 growth periods studied, two displayed exponential, 12 linear and 10 linear-plateau growth. There was no effect of FM on growth curve form. In May and June, LI increased with canopy height, up to 0.93. Stubble height and days of growth impacted FGR with an interaction. There was no treatment impact on root density. Weather caused variation in FGR. A low FGR risk occurs at high elevations; greater risk occurs east of the plateau.
The Effect of Endophytic Bacteria Bacillus subtilis and Salicylic Acid on Some Resistance and Quality Traits of Stored Solanum tuberosum L. Tubers Infected with Fusarium Dry Rot
Authors: Lastochkina Oksana , Pusenkova Liudmila , Garshina Darya , Yuldashev Ruslan , Shpirnaya Irina , Kasnak Cemal , Palamutoglu Recep , Mardanshin Ildar , Garipova Svetlana , Sobhani Mohammadhadi , Aliniaeifard Sasan
Published: June 2020

The effect of endophytic Bacillus subtilis (strains 10-4, 26D) and their compositions with salicylic acid (SA) on some resistance and quality traits of stored potatoes infected with Fusarium dry rot were studied. The experiments were carried out on hydroponically grown Solanum tuberosum L. tubers that were infected before storage with Fusarium oxysporum and coated with B. subtilis 10-4, 26D with and without exogenous SA, and then stored for six months. It has been shown that 10-4, 26D, 10-4 + SA, and 26D + SA reduced in different levels (up to 30–50%) the incidence of F. oxysporum-caused dry rot (with the highest effect for 10-4 + SA). SA notably enhanced the positive effect of 10-4, while for 26D, such an effect was not observed. All of the tested treatments increased amylase (AMY) and AMY inhibitors activity in infected tubers, while decreased Fusarium-induced protease activity (except in the case of 10-4 + SA, which promoted a slight increase) was revealed. 10-4, 26D, and their compositions with SA decreased (in different degrees) the pathogen-caused lipid peroxidation, proline, and reducing sugars accumulation in potatoes after long-term storage. It was also discovered 10-4 and 26D, regardless of SA presence, decrease pathogen-induced glycoalkaloids α-Solanine and α-Chaconine accumulation and preserved increased levels of starch and total dry matter in infected stored potatoes. The findings indicate endophytic B. subtilis and its compositions with SA is a promising eco-friendly and bio-safe approach to cope with postharvest decays of potato during long-term storage; however, when developing preparations-compositions it should take into account the strain-dependent manner of B. subtilis action together with SA.
A New Ferula (Apiaceae) Species from Southwest Anatolia: Ferula pisidica Akalın & Miski
Authors: Akalın Emine , Tuncay Hüseyin Onur , Olcay Bülent , Miski Mahmut
Published: June 2020

Ferula pisidica is a novel endemic species found in the vicinity of Karaman province of inner Mediterranean Region of Turkey. F. pisidica is morphologically distinct from F. haussknechtii and F. brevipedicellata by habit, sheaths, terminal leaf lobes, and mericarp size as well as by chemotaxonomic differences. The new species is described by morphological, carpological, ecological and phytochemical characteristics. Its relationships with the other related species and proposed conservation status will be reviewed.
Phytosociological Study, Diversity and Conservation Status of the Cloud Forest in the Dominican Republic
Authors: Cano Ortiz Ana , Musarella Carmelo Maria , Pinto Gomes Carlos José , Quinto Canas Ricardo , Piñar Fuentes José Carlos , Cano Eusebio
Published: June 2020

The study of the forest in rainy environments of the Dominican Republic reveals the presence of four types of vegetation formations, clearly differentiated from each other in terms of their floristic and biogeographical composition, and also significantly different from the rainforests of Cuba. This leads us to propose two new alliances and four plant associations located in northern mountain areas exposed to moisture-laden winds from the Atlantic: All. Rondeletio ochraceae-Clusion roseae (Ass. Cyatheo furfuracei-Prestoetum motanae; Ass. Ormosio krugii-Prestoetum montanae); and All. Rondeletio ochraceae-Didymopanion tremuli (Ass. Hyeronimo montanae-Magnolietum pallescentis; Hyeronimo dominguensis-Magnolietum hamorii). We pay special attention to the description of cloud forest types, since they have a high rate of endemic species, and therefore there are endemic habitats, which need special protective actions. Therefore, we apply the Shannon diversity index to characteristic, companion, non-endemic, and endemic species. As result, the association Ormosio krugii-Prestoetum montanae has a Shannon_T = 2.4 and a value of Shannon_E = 0, whereas the other 3 associations have a better conservation status with Shannon values in all cases > 0: This is due to a worse conservation status of the Eastern Cordillera, in comparison with the Central Cordillera and Sierra de Bhaoruco. Due to human activity, some areas are very poorly conserved, as evidenced by the diversity index and the presence of endemic tree and plant elements. The worst conserved in terms of the relationship between characteristic plants vegetation (cloud forest) in areas with high rainfall are in the Dominican Republic, along with its floristic diversity and state of conservation. This study has made it possible to significantly increase the botanical knowledge of this important habitat.
The Role of Microbial Inoculants on Plant Protection, Growth Stimulation, and Crop Productivity of the Olive Tree (Olea europea L.)
Authors: Bizos Georgios , Papatheodorou Efimia M. , Chatzistathis Theocharis , Ntalli Nikoletta , Aschonitis Vassilis G. , Monokrousos Nikolaos
Published: June 2020

The olive tree (Olea europaea L.) is an emblematic, long-living fruit tree species of profound economic and environmental importance. This study is a literature review of articles published during the last 10 years about the role of beneficial microbes [Arbuscular Mycorrhizal Fungi (AMF), Plant Growth Promoting Rhizobacteria (PGPR), Plant Growth Promoting Fungi (PGPF), and Endophytes] on olive tree plant growth and productivity, pathogen control, and alleviation from abiotic stress. The majority of the studies examined the AMF effect using mostly Rhizophagus irregularis and Glomus mosseae species. These AMF species stimulate the root growth improving the resistance of olive plants to environmental and transplantation stresses. Among the PGPR, the nitrogen-fixing bacteria Azospirillum sp. and potassium- and phosphorous-solubilizing Bacillus sp. species were studied extensively. These PGPR species were combined with proper cultural practices and improved considerably olive plant’s growth. The endophytic bacterial species Pseudomonas fluorescens and Bacillus sp., as well as the fungal species Trichoderma sp. were identified as the most effective biocontrol agents against olive tree diseases (e.g., Verticillium wilt, root rot, and anthracnose).
Genetic Characterization of an Endangered Chilean Endemic Species, Prosopis burkartii Muñoz, Reveals its Hybrids Parentage
Authors: Contreras Roberto , van den Brink Liesbeth , Burgos Boris , González Marlene , Gacitúa Sandra
Published: June 2020

The hybridization of Prosopis burkartii, a critically endangered endemic species, and the identification of its paternal species has not been genetically studied before. In this study we aimed to genetically confirm the origin of this species. To resolve the parental status of P. burkartii, inter-simple sequence repeat (ISSR), simple sequence repeats (SSR) and intron trnL molecular markers were used, and compared with Chilean species from the Algarobia and Strombocarpa sections. Out of seven ISSRs, a total of 70 polymorphic bands were produced in four species of the Strombocarpa section. An Multi-dimensional scaling (MDS) and Bayasian (STRUCTURE) analysis showed signs of introgression of genetic material in P. burkartii. Unweighted pair group method with arithmetic average (UPGMA) cluster analysis showed three clusters, and placed the P. burkartii cluster nested within the P. tamarugo group. Sequencing of the trnL intron showed a fragment of 535 bp and 529 bp in the species of the Algarobia and Strombocarpa sections, respectively. Using maximum parsimony (MP) and maximum likelihood (ML) trees with the trnL intron, revealed four clusters. A species-specific diagnostic method was performed, using the trnL intron Single Nucleotide Polymorphism (SNP). This method identified if individuals of P. burkartii inherited their maternal DNA from P. tamarugo or from P. strombulifera. We deduced that P. tamarugo and P. strombulifera are involved in the formation of P. burkartii.