bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Energy Drink Consumption on Physical Performance and Potential Danger of Inordinate Usage
Authors: Erdmann Jakub , Wiciński Michał , Wódkiewicz Eryk , Nowaczewska Magdalena , Słupski Maciej , Otto Stephan Walter , Kubiak Karol , Huk-Wieliczuk Elżbieta , Malinowski Bartosz
Published: July 2021

The rise in energy drink (ED) intake in the general population and athletes has been achieved with smart and effective marketing strategies. There is a robust base of evidence showing that adolescents are the main consumers of EDs. The prevalence of ED usage in this group ranges from 52% to 68%, whilst in adults is estimated at 32%. The compositions of EDs vary widely. Caffeine content can range from 75 to 240 mg, whereas the average taurine quantity is 342.28 mg/100 mL. Unfortunately, exact amounts of the other ED elements are often not disclosed by manufacturers. Caffeine and taurine in doses 3–6 mg/kg and 1–6 g, respectively, appear to be the main ergogenic elements. However, additive or synergic properties between them seem to be implausible. Because of non-unified protocol design, presented studies show inconsistency between ED ingestion and improved physical performance. Potential side effects caused by abusive consumption or missed contraindications are the aspects that are the most often overlooked by consumers and not fully elucidated by ED producers. In this review, the authors aimed to present the latest scientific information on ED components and their possible impact on improving physical performance as well as to bring emphasis to the danger of inordinate consumption.
Longitudinal Changes of Cytokines and Appetite in Older Hospitalized Patients
Authors: Pourhassan Maryam , Babel Nina , Sieske Lars , Westhoff Timm Henning , Wirth Rainer
Published: July 2021

There are few data on the longitudinal association of cytokine and appetite among older hospitalized patients. We aimed to investigate the impact of the changes of inflammatory cytokines on appetite in older hospitalized patients. A total of 191 patients (mean age 81.3 ± 6.6 years, 64% women) participated in this prospective longitudinal observational study. Appetite was evaluated using the Edmonton Symptom Assessment System on admission and after seven days. Serum cytokines such as IL-1β, IL-6, IL-8, IL-10, IL-12p70, IL-17, IL-18, IL-23 and IL-33, IFN-α2, IFN-γ, TNF-α and MCP-1 were measured both times. No significant differences in the mean serum levels of all the cytokines could be detected overtime in relation to appetite changes, except for IL-18. Appetite significantly deteriorated overtime in patients with increasing IL-18 levels and improved in those without significant changes in IL-18 levels. In a stepwise regression analysis, changes of IL-18 levels were the major independent predictor for the changes of patients’ appetite and explained 4% of the variance, whereas other cytokines and variables, such as age, sex, infection and disease, did not show any impact on appetite changes. We conclude that IL-18 seems to exert a significant impact on appetite in acutely ill older hospitalized patients and should, therefore, be considered as a potential target in the diagnosis, prevention and treatment of malnutrition.
Hepcidin and Iron Deficiency in Women One Year after Sleeve Gastrectomy: A Prospective Cohort Study
Authors: Lefebvre Thibaud , Coupaye Muriel , Esposito-Farèse Marina , Gault Nathalie , Talbi Neila , Quintin Caroline , Schmitt Caroline , Bekri Soumeya , Bado André , Puy Hervé , Msika Simon , Brasse-Lagnel Carole , Karim Zoubida
Published: July 2021

Iron deficiency with or without anemia, needing continuous iron supplementation, is very common in obese patients, particularly those requiring bariatric surgery. The aim of this study was to address the impact of weight loss on the rescue of iron balance in patients who underwent sleeve gastrectomy (SG), a procedure that preserves the duodenum, the main site of iron absorption. The cohort included 88 obese women; sampling of blood and duodenal biopsies of 35 patients were performed before and one year after SG. An analysis of the 35 patients consisted in evaluating iron homeostasis including hepcidin, markers of erythroid iron deficiency (soluble transferrin receptor (sTfR) and erythrocyte protoporphyrin (PPIX)), expression of duodenal iron transporters (DMT1 and ferroportin) and inflammatory markers. After surgery, sTfR and PPIX were decreased. Serum hepcidin levels were increased despite the significant reduction in inflammation. DMT1 abundance was negatively correlated with higher level of serum hepcidin. Ferroportin abundance was not modified. This study shed a new light in effective iron recovery pathways after SG involving suppression of inflammation, improvement of iron absorption, iron supply and efficiency of erythropoiesis, and finally beneficial control of iron homeostasis by hepcidin. Thus, recommendations for iron supplementation of patients after SG should take into account these new parameters of iron status assessment.
Data-Driven Methods for Spectator Symmetry Plane Estimation in CBM Experiment at FAIR
Authors: Golosov Oleg , Selyuzhenkov Ilya , Kashirin Evgeny
Published: July 2021

The Compressed Baryonic Matter experiment (CBM) at FAIR aims to study the area of the QCD phase diagram at high net baryon densities and moderate temperatures with collisions of heavy ions at sNN=2.8–4.9 GeV. The anisotropic transverse flow is one of the most important observable phenomena in a study of the properties of matter created in such collisions. Flow measurements require the knowledge of the collision symmetry plane, which can be determined from the deflection of the collision spectators in the plane transverse to the direction of the moving ions. The CBM performance for projectile spectator symmetry plane estimation is studied with GEANT4 Monte Carlo simulations using collisions of gold ions with beam momentum of 12A GeV/c generated with the DCM-QGSM-SMM model. Different data-driven methods to extract the correction factor in flow analysis for the resolution of the spectator symmetry plane estimated with the CBM Projectile Spectator Detector are investigated.
Bluetongue and Epizootic Hemorrhagic Disease in the United States of America at the Wildlife–Livestock Interface
Authors: Rivera Nelda A. , Varga Csaba , Ruder Mark G. , Dorak Sheena J. , Roca Alfred L. , Novakofski Jan E. , Mateus-Pinilla Nohra E.
Published: July 2021

Bluetongue (BT) and epizootic hemorrhagic disease (EHD) cases have increased worldwide, causing significant economic loss to ruminant livestock production and detrimental effects to susceptible wildlife populations. In recent decades, hemorrhagic disease cases have been reported over expanding geographic areas in the United States. Effective BT and EHD prevention and control strategies for livestock and monitoring of these diseases in wildlife populations depend on an accurate understanding of the distribution of BT and EHD viruses in domestic and wild ruminants and their vectors, the Culicoides biting midges that transmit them. However, national maps showing the distribution of BT and EHD viruses and the presence of Culicoides vectors are incomplete or not available at all. Thus, efforts to accurately describe the potential risk of these viruses on ruminant populations are obstructed by the lack of systematic and routine surveillance of their hosts and vectors. In this review, we: (1) outline animal health impacts of BT and EHD in the USA; (2) describe current knowledge of the distribution and abundance of BT and EHD and their vectors in the USA; and (3) highlight the importance of disease (BT and EHD) and vector surveillance for ruminant populations.
Genotypic Characterization of Clinical Isolates of Staphylococcus aureus from Pakistan
Authors: Khan Saeed , Marasa Bernard S. , Sung Kidon , Nawaz Mohamed
Published: July 2021

In this study, we compared pulsed-field gel electrophoretic (PFGE), multilocus sequence typing (MLST), Staphylococcal cassette chromosome mec (SCCmec), spa typing, and virulence gene profiles of 19 Panton–Valentine leucocidin (PVL)-positive, multidrug-, and methicillin-resistant clinical Staphylococcus aureus (MRSA) isolates obtained from a hospital intensive care unit in Pakistan. The isolates exhibited 10 pulsotypes, contained eight adhesin genes (bbp, clfA, clfB, cna, fnbA, fnbB, map-eap, and spa), 10 toxin genes (hla, hlb, hld, hlg, pvl, sed, see, seg, seh, and tst), and two other virulence genes (cfb, v8) that were commonly present in all isolates. The spa-typing indicated seven known spa types (t030, t064, t138, t314, t987, t1509, and t5414) and three novel spa types. MLST analysis indicated eight ST types (ST8, ST15, ST30, ST239, ST291, ST503, ST772, and ST1413). All isolates belonged to the agr group 1. Most of the isolates possessed SCCmec type III, but some isolates had it in combination with types SCCmec IV and V. The presence of multidrug-resistant MRSA isolates in Pakistan indicates poor hygienic conditions, overuse of antibiotics, and a lack of rational antibiotic therapy that have led to the evolution and development of hypervirulent MRSA clones. The study warrants development of a robust epidemiological screening program and adoption of effective measures to stop their spread in hospitals and the community.
Time-Course Transcriptome Profiling of a Poxvirus Using Long-Read Full-Length Assay
Authors: Tombácz Dóra , Prazsák István , Torma Gábor , Csabai Zsolt , Balázs Zsolt , Moldován Norbert , Dénes Béla , Snyder Michael , Boldogkői Zsolt
Published: July 2021

Viral transcriptomes that are determined using first- and second-generation sequencing techniques are incomplete. Due to the short read length, these methods are inefficient or fail to distinguish between transcript isoforms, polycistronic RNAs, and transcriptional overlaps and readthroughs. Additionally, these approaches are insensitive for the identification of splice and transcriptional start sites (TSSs) and, in most cases, transcriptional end sites (TESs), especially in transcript isoforms with varying transcript ends, and in multi-spliced transcripts. Long-read sequencing is able to read full-length nucleic acids and can therefore be used to assemble complete transcriptome atlases. Although vaccinia virus (VACV) does not produce spliced RNAs, its transcriptome has a high diversity of TSSs and TESs, and a high degree of polycistronism that leads to enormous complexity. We applied single-molecule, real-time, and nanopore-based sequencing methods to investigate the time-lapse transcriptome patterns of VACV gene expression.
MicroRNA-155 Modulates Macrophages’ Response to Non-Tuberculous Mycobacteria through COX-2/PGE2 Signaling
Authors: Yuan Zhihong , Prasla Zohra , Lee Frances Eun-Hyung , Bedi Brahmchetna , Sutliff Roy L. , Sadikot Ruxana T.
Published: July 2021

Non-tuberculous mycobacteria (NTM) have been recognized as a causative agent of various human diseases, including severe infections in immunocompromised patients, such as people living with HIV. The most common species identified is the Mycobacterium avium-intracellulare complex (MAI/MAC), accounting for a majority of infections. Despite abundant information detailing the clinical significance of NTM, little is known about host–pathogen interactions in NTM infection. MicroRNAs (miRs) serve as important post-transcriptional regulators of gene expression. Using a microarray profile, we found that the expression of miR-155 and cyclo-oxygenase 2 (COX-2) is significantly increased in bone-marrow-derived macrophages from mice and human monocyte-derived macrophages from healthy volunteers that are infected with NTM. Antagomir against miR-155 effectively suppressed expression of COX-2 and reduced Prostaglandin E2(PGE2) secretion, suggesting that COX-2/PGE2 expression is dependent on miR-155. Mechanistically, we found that inhibition of NF-κB activity significantly reduced miR-155/COX-2 expression in infected macrophages. Most importantly, blockade of COX-2, E-prostanoid receptors (EP2 and EP4) enhanced killing of MAI in macrophages. These findings provide novel mechanistic insights into the role of miR-155/COX-2/PGE2 signalling and suggest that induction of these pathways enhances survival of mycobacteria in macrophages. Defining host–pathogen interactions can lead to novel immunomodulatory therapies for NTM infections which are difficult to treat.
Development of Immunoassays for Detection of Francisella tularensis Lipopolysaccharide in Tularemia Patient Samples
Authors: Hannah Emily E. , Pandit Sujata G. , Hau Derrick , DeMers Haley L. , Robichaux Kayleigh , Nualnoi Teerapat , Dissanayaka Anjana , Arias-Umana Jose , Green Heather R. , Thorkildson Peter , Pflughoeft Kathryn J. , Gates-Hollingsworth Marcellene A. , Ozsurekci Yasemin , AuCoin David P.
Published: July 2021

Francisella tularensis is the causative agent of tularemia, a zoonotic bacterial infection that is often fatal if not diagnosed and treated promptly. Natural infection in humans is relatively rare, yet persistence in animal reservoirs, arthropod vectors, and water sources combined with a low level of clinical recognition make tularemia a serious potential threat to public health in endemic areas. F. tularensis has also garnered attention as a potential bioterror threat, as widespread dissemination could have devastating consequences on a population. A low infectious dose combined with a wide range of symptoms and a short incubation period makes timely diagnosis of tularemia difficult. Current diagnostic techniques include bacterial culture of patient samples, PCR and serological assays; however, these techniques are time consuming and require technical expertise that may not be available at the point of care. In the event of an outbreak or exposure a more efficient diagnostic platform is needed. The lipopolysaccharide (LPS) component of the bacterial outer leaflet has been identified previously by our group as a potential diagnostic target. For this study, a library of ten monoclonal antibodies specific to F. tularensis LPS were produced and confirmed to be reactive with LPS from type A and type B strains. Antibody pairs were tested in an antigen-capture enzyme-linked immunosorbent assay (ELISA) and lateral flow immunoassay format to select the most sensitive pairings. The antigen-capture ELISA was then used to detect and quantify LPS in serum samples from tularemia patients for the first time to determine the viability of this molecule as a diagnostic target. In parallel, prototype lateral flow immunoassays were developed, and reactivity was assessed, demonstrating the potential utility of this assay as a rapid point-of-care test for diagnosis of tularemia.
Evaluation of Potential Transfer of the Pathogen Saprolegnia parasitica between Farmed Salmonids and Wild Fish
Authors: Tedesco Perla , Saraiva Marcia , Sandoval-Sierra Jose Vladimir , Fioravanti Maria Letizia , Morandi Benedetto , Dieguez-Uribeondo Javier , van West Pieter , Galuppi Roberta
Published: July 2021

Saprolegnia infections are among the main parasitic diseases affecting farmed salmonids. The distribution and potential transfer of Saprolegnia spp. between farms and the natural environment has been scarcely investigated. Therefore, this work aimed to study the diversity and abundance of oomycete species in salmonid farms, tributary water, and effluent water systems. Four trout farms in Italy and two Atlantic salmon farms in Scotland were considered. In Italian farms, 532 isolates of oomycetes were obtained from fish and water, at upstream, inside, and downstream the farms. In Scottish farms, 201 oomycetes isolates were obtained from water outside the farm and from fish and water inside the farming units. Isolates were identified to the species level through amplification and sequencing of the ITS rDNA region. In Italy, S. parasitica was significantly more present in farmed than in wild fish, while in water it was more frequently isolated from the wild, particularly in effluent systems, not associated with more frequent isolation of S. parasitica in wild fish downstream the farm. In Scotland, S. parasitica was the most prevalent species isolated from fish, while isolates from water were mostly Pythium spp. with few S. parasitica isolates from upstream and downstream the farms.
IgA Vasculitis Complicated by Both CMV Reactivation and Tuberculosis
Authors: Mizerska-Wasiak Małgorzata , Winiarska Maria , Nogal Karolina , Cichoń-Kawa Karolina , Pańczyk-Tomaszewska Małgorzata , Małdyk Jadwiga
Published: July 2021

Immunoglobulin A (IgA) vasculitis is the most common systemic vasculitis in the pediatric population. We present the case of a patient with IgA vasculitis with nephritis who developed cytomegalovirus (CMV) infection followed by Mycobacterium tuberculosis infection. In the literature, there are a few cases of IgA nephropathy accompanied by reactivation of CMV or tuberculosis. To the best of our knowledge, this is the first reported case of IgA vasculitis complicated by both CMV reactivation and tuberculosis. It is important to detect infections in patients with IgA vasculitis because they can induce and exacerbate the symptoms of the disease. Effective antimicrobial treatment facilitates the management of proteinuria and slows down the decline of renal function. Immunosuppressive therapy is a risk factor for reactivation of latent infections and makes patients more susceptible to its generalized and complicated course. This can be prevented by actively screening for hidden sites of infection.
Metabolic Host–Microbiota Interactions in Autophagy and the Pathogenesis of Inflammatory Bowel Disease (IBD)
Authors: Dowdell Alexander S. , Colgan Sean P.
Published: July 2021

Inflammatory bowel disease (IBD) is a family of conditions characterized by chronic, relapsing inflammation of the gastrointestinal tract. IBD afflicts over 3 million adults in the United States and shows increasing prevalence in the Westernized world. Current IBD treatments center on modulation of the damaging inflammatory response and carry risks such as immunosuppression, while the development of more effective treatments is hampered by our poor understanding of the molecular mechanisms of IBD pathogenesis. Previous genome-wide association studies (GWAS) have demonstrated that gene variants linked to the cellular response to microorganisms are most strongly associated with an increased risk of IBD. These studies are supported by mechanistic work demonstrating that IBD-associated polymorphisms compromise the intestine’s anti-microbial defense. In this review, we summarize the current knowledge regarding IBD as a disease of defects in host–microbe interactions and discuss potential avenues for targeting this mechanism for future therapeutic development.
Innovations and Patent Trends in the Development of USFDA Approved Protein Kinase Inhibitors in the Last Two Decades
Authors: Imran Mohd. , Asdaq Syed Mohammed Basheeruddin , Khan Shah Alam , Unnikrishnan Meenakshi Dhanalekshmi , Alamri Abdulhakeem S. , Alsanie Walaa F. , Alhomrani Majid , Mohzari Yahya , Alrashed Ahmed , AlMotairi Mohammed , Alkhaldi Eman H. , Alorabi Abeer K. , Alshrari Ahmed Subeh , Tauseef Mohammad , Abida  , Alaqel Saleh I. , Alam Ozair , Bakht Md. Afroz
Published: July 2021

Protein kinase inhibitors (PKIs) are important therapeutic agents. As of 31 May 2021, the United States Food and Drug Administration (USFDA) has approved 70 PKIs. Most of the PKIs are employed to treat cancer and inflammatory diseases. Imatinib was the first PKI approved by USFDA in 2001. This review summarizes the compound patents and the essential polymorph patents of the PKIs approved by the USFDA from 2001 to 31 May 2021. The dates on the generic drug availability of the PKIs in the USA market have also been forecasted. It is expected that 19 and 48 PKIs will be genericized by 2025 and 2030, respectively, due to their compound patent expiry. This may reduce the financial toxicity associated with the existing PKIs. There are nearly 535 reported PKs. However, the USFDA approved PKIs target only about 10–15% of the total said PKs. As a result, there are still a large number of unexplored PKs. As the field advances during the next 20 years, one can anticipate that PKIs with many scaffolds, chemotypes, and pharmacophores will be developed.
[68Ga]Ga-PSMA-11: The First FDA-Approved 68Ga-Radiopharmaceutical for PET Imaging of Prostate Cancer
Authors: Hennrich Ute , Eder Matthias
Published: July 2021

For the positron emission tomography (PET) imaging of prostate cancer, radiotracers targeting the prostate-specific membrane antigen (PSMA) are nowadays used in clinical practice. Almost 10 years after its discovery, [68Ga]Ga-PSMA-11 has been approved in the United States by the Food and Drug Administration (FDA) as the first 68Ga-radiopharmaceutical for the PET imaging of PSMA-positive prostate cancer in 2020. This radiopharmaceutical combines the peptidomimetic Glu-NH-CO-NH-Lys(Ahx)-HBED-CC with the radionuclide 68Ga, enabling specific imaging of tumor cells expressing PSMA. Such a targeting approach may also be used for therapy planning as well as potentially for the evaluation of treatment response.
Direct Powder Extrusion 3D Printing of Praziquantel to Overcome Neglected Disease Formulation Challenges in Paediatric Populations
Authors: Boniatti Janine , Januskaite Patricija , Fonseca Laís B. da , Viçosa Alessandra L. , Amendoeira Fábio C. , Tuleu Catherine , Basit Abdul W. , Goyanes Alvaro , Ré Maria-Inês
Published: July 2021

For the last 40 years, praziquantel has been the standard treatment for schistosomiasis, a neglected parasitic disease affecting more than 250 million people worldwide. However, there is no suitable paediatric formulation on the market, leading to off-label use and the splitting of commercial tablets for adults. In this study, we use a recently available technology, direct powder extrusion (DPE) three-dimensional printing (3DP), to prepare paediatric Printlets™ (3D printed tablets) of amorphous solid dispersions of praziquantel with Kollidon® VA 64 and surfactants (Span™ 20 or Kolliphor® SLS). Printlets were successfully printed from both pellets and powders obtained from extrudates by hot melt extrusion (HME). In vitro dissolution studies showed a greater than four-fold increase in praziquantel release, due to the formation of amorphous solid dispersions. In vitro palatability data indicated that the printlets were in the range of praziquantel tolerability, highlighting the taste masking capabilities of this technology without the need for additional taste masking excipients. This work has demonstrated the possibility of 3D printing tablets using pellets or powder forms obtained by HME, avoiding the use of filaments in fused deposition modelling 3DP. Moreover, the main formulation hurdles of praziquantel, such as low drug solubility, inadequate taste, and high and variable dose requirements, can be overcome using this technology.
In Vitro Wound-Healing Properties of Water-Soluble Terpenoids Loaded on Halloysite Clay
Authors: Marinelli Lisa , Cacciatore Ivana , Eusepi Piera , Dimmito Marilisa Pia , Di Rienzo Annalisa , Reale Marcella , Costantini Erica , Borrego-Sánchez Ana , García-Villén Fátima , Viseras César , Morroni Gianluca , Fioriti Simona , Brescini Lucia , Di Stefano Antonio
Published: July 2021

Recently, mineral healing clays have gained much attention for wound-dressing applications. Here, we selected halloysite (HAL) clay as a biocompatible, non-toxic material that is useful as a drug delivery system to enhance the healing properties of water-soluble terpenoids 1-3 (T1-3). Terpenoids-loaded HAL clay (TH1-3) was prepared and characterized by adsorption equilibrium studies, X-ray powder diffraction (XRPD), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), Fourier-transform infrared (FTIR) spectroscopy, and release studies. The results reveal that T1-3 were adsorbed at the HAL surface with good efficiency. The prevalent mechanism of drug retention is due to the adsorption via electrostatic interactions between the cationic groups of the T1-3 and the HAL’s external surface. Release studies demonstrated that T3 was released in a higher percentage (>60%) compared to T1-2 (≈50%). Additionally, TH1-3 were assessed for their antimicrobial activity and capability to promote the re-epithelialization of scratched HaCat monolayers, through the time-kill test and the wound-healing assays, respectively. The results reveal that all the tested formulations were able to reduce the microbial growth after 1 h of incubation and that they ensured complete wound closure after 48 h. Furthermore, at the concentration of 1 µg/mL, TH3 exhibited 45% wound closure at 24 h, compared to TH1 (27%) and TH2 (30%), proving to be the best candidate in making the tissue-repair process easier and faster.
Experimental and Theoretical Study on Theobromine Solubility Enhancement in Binary Aqueous Solutions and Ternary Designed Solvents
Authors: Jeliński Tomasz , Stasiak Dawid , Kosmalski Tomasz , Cysewski Piotr
Published: July 2021

The solubility of theobromine was studied both experimentally and theoretically. The solubility was determined spectrophotometrically at 25 °C in neat organic solvents, aqueous binary mixtures, Natural Deep Eutectic Solvents (NADES) and ternary NADES mixtures with water. It was found that addition of water in unimolar proportions with some organic solvents increases theobromine solubility compared to neat solvents. Additionally, using NADES results in a solubility increase of the studied compound not only in relation to water but also DMSO. The addition of water (0.2 molar fraction) to NADES is responsible for an even larger increase of solubility. The measured solubilities were interpreted in terms of three theoretical frameworks. The first one—belonging to the set of data reduction techniques—proved to be very efficient in quantitative back-computations of excess solubility of theobromine in all studied systems. The default approach utilizing the well-recognized COSMO-RS (Conductor-like Screening Model for Real Solvents) framework offered at most a qualitative solubility description. The extended search for possible contacts provided evidence for the existence of many intermolecular complexes that alter the electron density of the solute molecule, thus influencing solubility computations. Taking into account such intermolecular contacts by using the COSMO-RS-DARE (Conductor-like Screening Model for Realistic Solvation-Dimerization, Aggregation, and Reaction Extension) framework seriously increased the accuracy of solubility computations.
Tailored Hydrogels as Delivery Platforms for Conditioned Medium from Mesenchymal Stem Cells in a Model of Acute Colitis in Mice
Authors: Sendon-Lago Juan , Rio Lorena Garcia-del , Eiro Noemi , Diaz-Rodriguez Patricia , Avila Leandro , Gonzalez Luis O. , Vizoso Francisco J. , Perez-Fernandez Roman , Landin Mariana
Published: July 2021

Inflammatory bowel disease (IBD), including Crohn’s disease (CD) and ulcerative colitis (UC), is increasingly prevalent and current therapies are not completely effective. Mesenchymal stem cells are emerging as a promising therapeutic option. Here, the effect of local hydrogel application loaded with conditioned medium (CM) from human uterine cervical stem cells (hUCESC-CM) in an experimental acute colitis mice model has been evaluated. Colitis induction was carried out in C57BL/6 mice by dissolving dextran sulfate sodium (DSS) in drinking water for nine days. Ulcers were treated by rectal administration of either mesalazine (as positive control) or a mucoadhesive and thermosensitive hydrogel loaded with hUCESC-CM (H-hUCESC-CM). Body weight changes, colon length, and histopathological analysis were evaluated. In addition, pro-inflammatory TNF-α, IL-6, and IFN-γ mRNA levels were measured by qPCR. Treatment with H-hUCESC-CM inhibited body weight loss and colon shortening and induced a significant decrease in colon mucosa degeneration, as well as TNF-α, IFN-γ, and IL-6 mRNA levels. Results indicate that H-hUCESC-CM effectively alleviated DSS-induced colitis in mice, suggesting that H-hUCESC-CM may represent an attractive cell-free therapy for local treatment of IBD.
Freeze-Dried Mesenchymal Stem Cell-Secretome Pharmaceuticalization: Optimization of Formulation and Manufacturing Process Robustness
Authors: Mocchi Michela , Bari Elia , Marrubini Giorgio , Bonda Andrea Foglio , Perteghella Sara , Tartara Fulvio , Cofano Fabio , Perna Giuseppe di , Giovannelli Lorella , Mandracchia Delia , Sorlini Marzio , Garbossa Diego , Torre Maria Luisa , Segale Lorena
Published: July 2021

Producing mesenchymal stem cell (MSC)-secretome for dose escalation studies and clinical practice requires scalable and good manufacturing practice (GMP)-compliant production procedures and formulation into a standardized medicinal product. Starting from a method that combines ultrafiltration and freeze-drying to transform MSC-secretome into a pharmaceutical product, the lyosecretome, this work aims to: (i) optimize the lyosecretome formulation; (ii) investigate sources of variability that can affect the robustness of the manufacturing process; (iii) modify the ultrafiltration step to obtain a more standardized final product. Design of experiments and principal component analysis of the data were used to study the influence of batch production, lyophilization, mannitol (M)/sucrose (S) binary mixture, selected as cryoprotectant excipients, and the total amount of excipients on the extracellular vesicles (EV) particle size, the protein and lipid content and the in vitro anti-elastase. The different excipients ratios did not affect residual moisture or EV particle size; simultaneously, proteins and lipids were better preserved in the freeze-dried product using the maximum total concentration of excipients (1.5% w/v) with a M:S ratio of about 60% w/w. The anti-elastase activity was instead better preserved using 0.5% w/w of M as excipient. The secretome batch showed to be the primary source of variability; therefore, the manufacturing process has been modified and then validated: the final product is now concentrated to reach a specific protein (and lipid) concentration instead of cell equivalent concentration. The new standardization approach led to a final product with more reproducible quali-quantitative composition and higher biological activity.
Investigation of Tactile Illusion Based on Gestalt Theory
Authors: Komura Hiraku , Nakamura Toshiki , Ohka Masahiro
Published: July 2021

Time-evolving tactile sensations are important in communication between animals as well as humans. In recent years, this research area has been defined as “tactileology,” and various studies have been conducted. This study utilized the tactile Gestalt theory to investigate these sensations. Since humans recognize shapes with their visual sense and melodies with their auditory sense based on the Prägnanz principle in the Gestalt theory, this study assumed that a time-evolving texture sensation is induced by a tactile Gestalt. Therefore, the operation of such a tactile Gestalt was investigated. Two psychophysical experiments were conducted to clarify the operation of a tactile Gestalt using a tactile illusion phenomenon called the velvet hand illusion (VHI). It was confirmed that the VHI is induced in a tactile Gestalt when the laws of closure and common fate are satisfied. Furthermore, it was clarified that the tactile Gestalt could be formulated using the proposed factors, which included the laws of elasticity and translation, and it had the same properties as a visual Gestalt. For example, the strongest Gestalt factor had the highest priority among multiple competing factors.
Effects of Scarification, Phytohormones, Soil Type, and Warming on the Germination and/or Seedling Performance of Three Tamaulipan Thornscrub Forest Species
Authors: Luera Paula , Wahl-Villarreal Kimberly , Christoffersen Bradley O. , Treviño Abeny , Soti Pushpa , Gabler Christopher A.
Published: July 2021

The Tamaulipan thornforests of south Texas and northeast Mexico are an ecologically and economically important conservation hotspot. Thornforest restoration is limited by native tree and shrub seedling availability for planting. Seedling shortages arise from low seed availability and knowledge gaps regarding best practices for germinating and growing the 70+ thornforest species desired for restoration plantings. To fill key knowledge gaps, we investigated three ecologically important thornforest species with low or highly variable germination or seedling survival rates: Ebenopsis ebano, Cordia boissieri, and Zanthoxylum fagara. For each, we quantified the effects of different dosages of chemical seed treatments used to promote germination (sulfuric acid, SA; gibberellic acid, GA; indole-3-butyric acid, IBA) on germination likelihood and timing. We also quantified the effects that these chemical seed treatments, soil media mixture type, and soil warming had on seedling survival, growth, and root morphology. Ebenopsis germination peaked (>90%) with 40–60 min SA treatment. Cordia germination peaked (40%) with 100 mg/L GA treatment. Zanthoxylum germination was negligible across all treatments. Seed molding was rare but stirring during SA treatment reduced Ebenopsis molding by 4%. Ebenopsis seedling survival, height, leaf count, and root morphology were minimally affected by seed treatments, generally reduced by warming, and influenced by soil mix, which also mediated responses to warming. These results suggest improvements to existing practices that could increase Ebenopsis germination by 10–20% and potentially double Cordia germination.
In Vitro Production and Exudation of 20-Hydroxymaytenin from Gymnosporia heterophylla (Eckl. and Zeyh.) Loes. Cell Culture
Authors: Pitakbut Thanet , Spiteller Michael , Kayser Oliver
Published: July 2021

The metabolite 20-Hydroxymaytenin (20-HM) is a member of the quinone-methide pentacyclic triterpenoids (QMTs) group. This metabolite group is present only in Celastraceae plants, and it has shown various biological activities from antioxidant to anticancer properties. However, most QMTs metabolites including 20-HM cannot be synthesized in a laboratory. Therefore, we optimized a plant tissue culture protocol and examined the potential of Gymnosporia heterophylla (synonym. Maytenus heterophylla) to produce 20-HM in an in vitro experiment. For the first time, we reported the optimum callus induction medium with a high percentage success rate of 82% from the combination of 1 mg/L indole-3-butyric acid and 5 mg/L naphthalene acetic acid. Later, our cell suspension culture cultivated in the optimum medium provided approximately 0.35 mg/g fresh weight of 20-HM. This concentration is roughly 87.5 times higher than a concentration of 20-HM presenting in Elaeodendron croceum (Celastraceae) leaves. In addition, we also found that 20-HM presented in a cultivation medium, suggesting that G. heterophylla cells secreted 20-HM as an exudate in our experiment. Noticeably, 20-HM was missing when Penicillium cf. olsonii occurred in the medium. These findings hint at an antifungal property of 20-HM.
Growth Rate, Dry Matter Accumulation, and Partitioning in Soybean (Glycine max L.) in Response to Defoliation under High-Rainfall Conditions
Authors: Raza Muhammad Ali , Gul Hina , Yang Feng , Ahmed Mukhtar , Yang Wenyu
Published: July 2021

The frequency of heavy rains is increasing with climate change in regions that already have high annual rainfall (i.e., Sichuan, China). Crop response under such high-rainfall conditions is to increase dry matter investment in vegetative parts rather than reproductive parts. In the case of soybean, leaf redundancy prevails, which reduces the light transmittance and seed yield. However, moderate defoliation of soybean canopy could reduce leaf redundancy and improve soybean yield, especially under high-rainfall conditions. Therefore, the effects of three defoliation treatments (T1, 15%; T2, 30%; and T3, 45% defoliation from the top of the soybean canopy; defoliation treatments were applied at the pod initiation stage of soybean) on the growth and yield parameters of soybean were evaluated through field experiments in the summer of 2017, 2018, and 2019. All results were compared with nondefoliated soybean plants (CK) under high-rainfall conditions. Compared with CK, treatment T1 significantly (p < 0. 05) improved the light transmittance and photosynthetic rate of soybean. Consequently, the leaf greenness was enhanced by 22%, which delayed the leaf senescence by 13% at physiological maturity. Besides, compared to CK, soybean plants achieved the highest values of crop growth rate in T1, which increased the total dry matter accumulation (by 6%) and its translocation to vegetative parts (by 4%) and reproductive parts (by 8%) at physiological maturity. This improved soybean growth and dry matter partitioning to reproductive parts in T1 enhanced the pod number (by 23%, from 823.8 m−2 in CK to 1012.7 m−2 in T1) and seed number (by 11%, from 1181.4 m−2 in CK to 1311.7 m−2 in T1), whereas the heavy defoliation treatments considerably decreased all measured growth and yield parameters. On average, treatment T1 increased soybean seed yield by 9% (from 2120.2 kg ha−1 in CK to 2318.2 kg ha−1 in T1), while T2 and T3 decreased soybean seed yield by 19% and 33%, respectively, compared to CK. Overall, these findings indicate that the optimum defoliation, i.e., T1 (15% defoliation), can decrease leaf redundancy and increase seed yield by reducing the adverse effects of mutual shading and increasing the dry matter translocation to reproductive parts than vegetative parts in soybean, especially under high-rainfall conditions. Future studies are needed to understand the internal signaling and the molecular mechanism controlling and regulating dry matter production and partitioning in soybean, especially from the pod initiation stage to the physiological maturity stage.
Selenium Biofortification of Soybean Seeds Influences Physiological Responses of Seedlings to Osmotic Stress
Authors: Galić Lucija , Špoljarević Marija , Jakovac Elizabeta , Ravnjak Boris , Teklić Tihana , Lisjak Miroslav , Perić Katarina , Nemet Franjo , Lončarić Zdenko
Published: July 2021

Climate change poses a serious threat to agricultural production. Water deficit in agricultural soils is one of the consequences of climate change that has a negative impact on crop growth and yield. Selenium (Se) is known to be involved in plant defense against biotic and abiotic stress through metabolic, structural, and physiological activity in higher plants. The aim of this study was to investigate the physiological response of Se-biofortified soybean (Glycine max (L.) Merrill) seedlings under osmotic stress. For this research, we used biofortified soybean grain obtained after foliar Se biofortification in 2020. The experiment was conducted in a growth chamber with two cultivars (Lucija and Sonja) grown on filter paper in three replicates. The experiment was carried out with two watering treatments: distilled water (PEG-0) and 2.5% polyethylene glycol 6000 (PEG-2.5) on Se-biofortified seeds (Se) and nonbiofortified seeds (wSe). Contents of lipid peroxidation product (LP), free proline (PRO), total phenolic content (TP), ferric reducing antioxidant power (FRAP), and ascorbic acid (AA) were analyzed in 7-days-old seedlings. Significant differences were detected in the Se content of soybean grains between the two cultivars. A milder reaction to PEG-2.5 was observed in cultivar Lucija in both Se and wSe treatments, which might represent the mitigating effects of Se on osmotic stress in this cultivar. Contrarily, in cultivar Sonja, Se adversely affected all analyzed traits in the PEG-2.5 treatment. Ultimately, Se is a pro-oxidant in Sonja, whereas it represents an anti-oxidant in Lucija. In conclusion, different soybean cultivars show contrasting physiological reactions to both osmotic stress and Se. However, the activation of antioxidant pathways in Sonja can also be interpreted as added value in soybean seedlings as a functional food.
Binding Properties of Photosynthetic Herbicides with the QB Site of the D1 Protein in Plant Photosystem II: A Combined Functional and Molecular Docking Study
Authors: Battaglino Beatrice , Grinzato Alessandro , Pagliano Cristina
Published: July 2021

Photosystem II (PSII) is a multi-subunit enzymatic complex embedded in the thylakoid membranes responsible for the primary photosynthetic reactions vital for plants. Many herbicides used for weed control inhibit PSII by interfering with the photosynthetic electron transport at the level of the D1 protein, through competition with the native plastoquinone for the QB site. Molecular details of the interaction of these herbicides in the D1 QB site remain to be elucidated in plants. Here, we investigated the inhibitory effect on plant PSII of the PSII-inhibiting herbicides diuron, metobromuron, bentazon, terbuthylazine and metribuzin. We combined analysis of OJIP chlorophyll fluorescence kinetics and PSII activity assays performed on thylakoid membranes isolated from pea plants with molecular docking using the high-resolution PSII structure recently solved from the same plant. Both approaches showed for terbuthylazine, metribuzin and diuron the highest affinity for the D1 QB site, with the latter two molecules forming hydrogen bonds with His215. Conversely, they revealed for bentazon the lowest PSII inhibitory effect accompanied by a general lack of specificity for the QB site and for metobromuron an intermediate behavior. These results represent valuable information for future design of more selective herbicides with enhanced QB binding affinities to be effective in reduced amounts.
Diversity Assessment of the Montenegrin Maize Landrace Gene Pool Maintained in Two Gene Banks
Authors: Babić Vojka , Andjelkovic Violeta , Jovovic Zoran , Babic Milosav , Vasic Vladimir , Kravic Natalija
Published: July 2021

Due to the loss of agro-biodiversity, there is a strong effort to find apparent and efficient mechanisms for the conservation and sustainable use of genetic diversity. A joint monitoring of the diversity and collections structure of the Montenegrin maize landraces conserved in the Serbian (MRIZPGB) and Montenegrin (MGB) gene banks has been conducted in order to improve the composition of the collections and to identify and eliminate possible redundancy. Based on a separate analysis of white- and yellow-orange maize landraces, it can be concluded that the diversity and evolution of distinct maize landraces grown and collected in Montenegro have been simultaneously shaped by both environmental (i.e., natural selection) and socially driven factors (farmers’ selection, migration and colonization processes of the human population). Although it has been determined that the authenticity and variability of the Montenegrin maize landraces gene pool have largely been preserved in the MRIZPGB collection, a significant amount of redundancy was observed. The obtained results will contribute to the cost-efficient conservation of the maize gene pool in the Montenegrin and Serbian gene banks. The recognized and well-preserved original variability of the MRIZPGB and MGB Montenegrin gene pool represents a valuable source for pre-breeding activities on broadening the white and flint maize breeding programmes under temperate conditions.
Importance of Plants with Extremely Small Populations (PSESPs) in Endemic-Rich Areas, Elements Often Forgotten in Conservation Strategies
Authors: Cogoni Donatella , Fenu Giuseppe , Dessì Carlo , Deidda Angela , Giotta Cesario , Piccitto Marcello , Bacchetta Gianluigi
Published: July 2021

The distribution of the threatened fern Ophioglossum vulgatum L., a plant with extremely small populations (PSESPs) in Sardinia, is characterized by small disjunct populations with only a few individuals, and little is known about its status in the wild. To provide information for the conservation of O. vulgatum and with the aim to develop an in situ conservation strategy, we investigated its distribution, population size, and habitat. Field surveys confirmed that the species grows in only five localities. Two representative populations were selected for this study (Funtanamela and Gedili), and in each population, all plants were mapped and monitored monthly from April to August over an 8-year period. During the study, the populations had a very low number of reproductive plants and the populations appeared to be in decline, with the total number of plants per population slightly decreased in Gedili while a sharp reduction was recorded in Funtanamela due to wild boar threat. A fence was built in order to protect the site from further damage, but no noticeable signals of recovery were observed. The most urgent conservation requirement for this species is to preserve the threatened habitat of the remnant populations. Further field surveys and research are also required for an improved understanding of the species’ status.
Winter Frosts Reduce Flower Bud Survival in High-Mountain Plants
Authors: Wagner Johanna , Gruber Karla , Ladinig Ursula , Buchner Othmar , Neuner Gilbert
Published: July 2021

At higher elevations in the European Alps, plants may experience winter temperatures of −30 °C and lower at snow-free sites. Vegetative organs are usually sufficiently frost hardy to survive such low temperatures, but it is largely unknown if this also applies to generative structures. We investigated winter frost effects on flower buds in the cushion plants Saxifraga bryoides L. (subnival-nival) and Saxifraga moschata Wulfen (alpine-nival) growing at differently exposed sites, and the chionophilous cryptophyte Ranunculus glacialis L. (subnival-nival). Potted plants were subjected to short-time (ST) and long-time (LT) freezing between −10 and −30 °C in temperature-controlled freezers. Frost damage, ice nucleation and flowering frequency in summer were determined. Flower bud viability and flowering frequency decreased significantly with decreasing temperature and exposure time in both saxifrages. Already, −10 °C LT-freezing caused the first injuries. Below −20 °C, the mean losses were 47% (ST) and 75% (LT) in S. bryoides, and 19% (ST) and 38% (LT) in S. moschata. Winter buds of both saxifrages did not supercool, suggesting that damages were caused by freeze dehydration. R. glacialis remained largely undamaged down to −30 °C in the ST experiment, but did not survive permanent freezing below −20 °C. Winter snow cover is essential for the survival of flower buds and indirectly for reproductive fitness. This problem gains particular relevance in the context of winter periods with low precipitation and winter warming events leading to the melting of the protective snowpack.