bluuebunny/crossref_metadata_2025 huggingface dataset:
Ecoepidemiology and Potential Transmission of Vibrio cholerae among Different Environmental Niches: An Upcoming Threat in Egypt
Authors: Ismail Eman M. , Kadry Mona , Elshafiee Esraa A. , Ragab Eman , Morsy Eman A. , Rizk Omar , Zaki Manal M.
Published: February 2021

Cholera is a negative public health event caused by Vibrio cholerae. Although V. cholerae is abundant in natural environments, its pattern and transmission between different niches remain puzzling and interrelated. Our study aimed to investigate the occurrence of nonpathogenic V. cholerae in the natural environment during endemicity periods. It also aimed to highlight the role of molecular ecoepidemiology in mapping the routes of spread, transmission, and prevention of possible future cholera outbreaks. V. cholerae was detected in different aquatic environments, waterfowl, and poultry farms located along the length of the Nile River in Giza, Cairo, and Delta provinces, Egypt. After polymerase chain reaction amplification of the specific target outer membrane gene (Omp W) of suspected isolates, we performed sequence analysis, eventually using phylogenetic tree analysis to illustrate the possible epidemiological relationships between different sequences. Data revealed a significant variation in the physicochemical conditions of the examined Nile districts related to temporal, spatial, and anthropogenic activities. Moreover, data showed an evident association between V. cholerae and the clinically diseased Synodontis schall fish. We found that the environmental distress triggered by the salinity shift and elevated temperature in the Middle Delta of the Nile River affects the pathogenesis of V. cholerae, in addition to the characteristics of fish host inhabiting the Rosetta Branch at Kafr El-Zayat, El-Gharbia province, Egypt. In addition, we noted a significant relationship between V. cholerae and poultry sources that feed on the Nile dikes close to the examined districts. Sequence analysis revealed clustering of the waterfowl and broiler chicken isolates with human and aquatic isolated sequences retrieved from the GenBank databases. From the obtained data, we hypothesized that waterfowl act as a potential vector for the intermediate transmission of cholera. Therefore, continuous monitoring of Nile water quality and mitigation of Nile River pollution, in addition to following good managemental practices (GMPs), general hygienic guidelines, and biosecurity in the field of animal production and industry, might be the way to break this cyclic transmission between human, aquatic, and animal sectors.
Development of a Multiplex PCR and Magnetic DNA Capture Assay for Detecting Six Species Pathogens of the Genera Anaplasma and Ehrlichia in Canine, Bovine, Caprine and Ovine Blood Samples from Grenada, West Indies
Authors: Sharma Bhumika , Ganta Roman R. , Stone Diana , Alhassan Andy , Lanza-Perea Marta , Matthew Belmar Vanessa , Karasek Inga , Cooksey Elizabeth , Butler Catherine M. , Gibson Kathryn , Wilkerson Melinda J.
Published: February 2021

Infections with tick-borne pathogens belonging to Anaplasma/Ehrlichia in various vertebrate hosts are a persistent problem resulting in nonspecific clinical signs during early infection. Diagnosis of single and multi-infections with these pathogens, causing diseases in companion/agricultural animals and people, remains a challenge. Traditional methods of diagnosis, such as microscopy and serology, have low sensitivity and specificity. Polymerase chain reaction (PCR) assays are widely used to detect early-phase infections, since these have high sensitivity and specificity. We report the development and validation of an assay involving PCR followed by magnetic capture method using species-specific oligonucleotides to detect six Anaplasma/Ehrlichia species pathogens in canine, bovine, caprine, and ovine blood samples. Overall, the assay application to 455 samples detected 30.1% (137/455) positives for one or more out of six screened pathogens. Single-pathogen infections were observed in 94.9% (130/137) of the positive samples, while co-infections were detected in 5.1% (7/137). Anaplasma marginale infection in cattle had the highest detection rate (34.4%), followed by canines positive for Anaplasma platys (16.4%) and Ehrlichia canis (13.9%). The assay aided in documenting the first molecular evidence for A. marginale in cattle and small ruminants and Ehrlichia chaffeensis and Ehrlichia ewingii in dogs in the Caribbean island of Grenada.
Evaluation of Two Methods to Concentrate SARS-CoV-2 from Untreated Wastewater
Authors: Dumke Roger , de la Cruz Barron Magali , Oertel Reinhard , Helm Björn , Kallies Rene , Berendonk Thomas U. , Dalpke Alexander
Published: February 2021

Use of wastewater-based epidemiology as a tool to record and manage the course of SARS-CoV-2 infections in human populations requires information about the efficiency of methods to concentrate the virus from wastewater. In the present study, we spiked untreated wastewater with quantified SARS-CoV-2 positive clinical material and enriched the virus by polyethylene glycol precipitation and ultrafiltration with Vivaspin 10 kDa MWCO columns. SARS-CoV-2 was detected and quantified by reverse transcription quantitative PCR (E- and S-gene) and droplet digital PCR. The concentration of virus with precipitation resulted in mean recoveries between 59.4% and 63.7% whereas rates from 33.0% to 42.6% after ultrafiltration of samples were demonstrated. The results suggest that the use of both methods allows an effective and practicable enrichment of SARS-CoV-2 from raw wastewater.
Whole Genome Sequencing and Antimicrobial Resistance of Staphylococcus aureus from Surgical Site Infections in Ghana
Authors: Egyir Beverly , Bentum Jeannette , Attram Naiki , Fox Anne , Obeng-Nkrumah Noah , Appiah-Korang Labi , Behene Eric , Kumordjie Selassie , Yeboah Clara , Agbodzi Bright , Bentil Ronald Essah , Tagoe Rhodalyn , Kofi Adu Tabi Blessing , Owusu Felicia , Dayie Nicholas T. K. D. , Donkor Eric S. , Nsaful Josephine , Asah-Opoku Kwaku , Nyarko Edward , Asumanu Edward , Larsen Anders Rhod , Wolfe David M. , Letizia Andrew G.
Published: February 2021

Staphylococcus aureus (S. aureus) is a common cause of surgical site infections (SSIs) globally. Data on the occurrence of methicillin-susceptible S. aureus (MSSA) as well as methicillin-resistant S. aureus (MRSA) among patients with surgical site infections (SSIs) in sub-Saharan African are scarce. We characterized S. aureus from SSIs in Ghana using molecular methods and antimicrobial susceptibility testing (AST). Wound swabs or aspirate samples were collected from subjects with SSIs. S. aureus was identified by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF-MS); AST was performed by Kirby-Bauer disk diffusion, and results were interpreted according to the Clinical and Laboratory Standards Institute (CLSI) guideline. Detection of spa, mecA, and pvl genes was performed by polymerase chain reaction (PCR). Whole-genome sequencing (WGS) was done using the Illumina MiSeq platform. Samples were collected from 112 subjects, with 13 S. aureus isolates recovered. Of these, 92% were sensitive to co-trimoxazole, 77% to clindamycin, and 54% to erythromycin. Multi-drug resistance was detected in 5 (38%) isolates. The four mecA gene-positive MRSA isolates detected belonged to ST152 (n = 3) and ST5 (n = 1). In total, 62% of the isolates were positive for the Panton-Valentine leukocidin (pvl) toxin gene. This study reports, for the first time, a pvl-positive ST152-t355 MRSA clone from SSIs in Ghana. The occurrence of multi-drug-resistant S. aureus epidemic clones suggests that continuous surveillance is required to monitor the spread and resistance trends of S. aureus in hospital settings in the country.
Amyloid Beta Peptides and Th1 Cytokines Modulate Human Brain Vascular Smooth Muscle Tonic Contractile Capacity In Vitro: Relevance to Alzheimer’s Disease?
Authors: Yun J. Winny , Washington Caretia , McCormick Joi , Stevenson Emily , Alexander J. Steven
Published: February 2021

Alzheimer’s Disease (AD) is a neurodegenerative condition characterized both by the presence of tau protein neurofibrillary tangles and amyloid beta (Aβ) containing extracellular “plaques”. The cleavage of amyloid precursor protein (APP) yields several Aβ peptides. Although Aβ toxicity to neurons has been described extensively, its effects on other components of the neurovasculature such as vascular smooth muscle cells have been less well characterized. AD is now also recognized as a neurovascular disease characterized by cerebral microbleeds and disturbances in autoregulation. AD is also a neuroinflammatory condition in which several proinflammatory cytokines are elevated and may contribute to the intensification of AD severity. Cerebral autoregulation (the mechanism by which brain blood flow is maintained despite changes in perfusion pressure) is extremely tightly controlled in the brain and shows disturbances in AD. The failure of autoregulation in AD may make the brain susceptible to cerebral microbleeds through a reduced capacity to limit blood flow when pressure is increased. Conversely, reduced vasodilation during low flow might could also exacerbate tissue hypoxia. Currently, whether and how Aβ peptides and inflammatory cytokines depress brain smooth muscle cell tonic contraction is not known, but could reveal important targets in the preservation of autoregulation which is disturbed in AD. We used a collagen gel contractility assay to evaluate the influence of Aβ25-35, Aβ1-40 and Aβ1-42 peptides and inflammatory cytokines on the tonic contractility of human brain vascular smooth muscle cells (HBVSMC) as an in vitro model of cerebral autoregulation. We found that 5 and 10 μM Aβ1-42 significantly depressed HBVSM contractility, while Aβ1-40 5–20 μM had no effect on contractility. Conversely, Aβ25-35 (1–50 μM) increased contractility. Interestingly, the inflammatory cytokines TNF-α (20 ng/mL), IL-1β (20 ng/mL) and IFN-γ (1000 U/mL) also depressed HBVSM tonic contractility alone and in combination. These data suggest that both the inflammatory milieu in AD as well as the abundance of Aβ peptides may promote autoregulatory failure and increase brain susceptibility to dysregulated perfusion and microbleeds which are an important and devastating characteristic of AD.
Novel Potent and Selective DPP-4 Inhibitors: Design, Synthesis and Molecular Docking Study of Dihydropyrimidine Phthalimide Hybrids
Authors: Mourad Ahmed A. E. , Khodir Ahmed E. , Saber Sameh , Mourad Mai A. E.
Published: February 2021

Background: Dipeptidyl peptidase-4 (DPP-4) inhibitors have emerged as anti-hyperglycemic agents that improve glycemic control in type 2 diabetic patients, either as monotherapy or in combination with other antidiabetic drugs. Methods: A novel series of dihydropyrimidine phthalimide hybrids was synthesized and evaluated for their in vitro and in vivo DPP-4 inhibition activity and selectivity using alogliptin as reference. Oral glucose tolerance test was assessed in type 2 diabetic rats after chronic treatment with the synthesized hybrids ± metformin. Cytotoxicity and antioxidant assays were performed. Additionally, molecular docking study with DPP-4 and structure activity relationship of the novel hybrids were also studied. Results: Among the synthesized hybrids, 10g, 10i, 10e, 10d and 10b had stronger in vitro DPP-4 inhibitory activity than alogliptin. Moreover, an in vivo DPP-4 inhibition assay revealed that 10g and 10i have the strongest and the most extended blood DPP-4 inhibitory activity compared to alogliptin. In type 2 diabetic rats, hybrids 10g, 10i and 10e exhibited better glycemic control than alogliptin, an effect that further supported by metformin combination. Finally, 10j, 10e, 10h and 10d had the highest radical scavenging activity in DPPH assay. Conclusions: Hybrids 10g, 10i and 10e are potent DPP-4 inhibitors which may be beneficial for T2DM treatment.
Modulation of the Serotonergic Receptosome in the Treatment of Anxiety and Depression: A Narrative Review of the Experimental Evidence
Authors: Villas-Boas Gustavo R. , Lavorato Stefânia N. , Paes Marina M. , de Carvalho Pablinny M. G. , Rescia Vanessa C. , Cunha Mila S. , de Magalhães-Filho Manoel F. , Ponsoni Luis F. , de Carvalho Adryano Augustto Valladao , de Lacerda Roseli B. , da S. Leite Lais , da S. Tavares-Henriques Matheus , Lopes Luiz A. F. , Oliveira Luiz G. R. , Silva-Filho Saulo E. , da Silveira Ana P. S. , Cuman Roberto K. N. , de S. Silva-Comar Francielli M. , Comar Jurandir F. , do A. Brasileiro Luana , dos Santos Jussileide N. , de Freitas William R. , Leão Katyuscya V. , da Silva Jonatas G. , Klein Raphael C. , Klein Mary H. F. , da S. Ramos Bruno H. , Fernandes Cristiane K. C. , de L. Ribas Dayane G. , Oesterreich Silvia A.
Published: February 2021

Serotonin (5-HT) receptors are found throughout central and peripheral nervous systems, mainly in brain regions involved in the neurobiology of anxiety and depression. 5-HT receptors are currently promising targets for discovering new drugs for treating disorders ranging from migraine to neuropsychiatric upsets, such as anxiety and depression. It is well described in the current literature that the brain expresses seven types of 5-HT receptors comprising eighteen distinct subtypes. In this article, we comprehensively reviewed 5-HT1-7 receptors. Of the eighteen 5-HT receptors known today, thirteen are G protein-coupled receptors (GPCRs) and represent targets for approximately 40% of drugs used in humans. Signaling pathways related to these receptors play a crucial role in neurodevelopment and can be modulated to develop effective therapies to treat anxiety and depression. This review presents the experimental evidence of the modulation of the “serotonergic receptosome” in the treatment of anxiety and depression, as well as demonstrating state-of-the-art research related to phytochemicals and these disorders. In addition, detailed aspects of the pharmacological mechanism of action of all currently known 5-HT receptor families were reviewed. From this review, it will be possible to direct the rational design of drugs towards new therapies that involve signaling via 5-HT receptors.
Endostatin Genetically Engineered Placental Mesenchymal Stromal Cells Carrying Doxorubicin-Loaded Mesoporous Silica Nanoparticles for Combined Chemo- and Antiangiogenic Therapy
Authors: de la Torre Paz , Paris Juan L. , Fernández-de la Torre Miguel , Vallet-Regí María , Flores Ana I.
Published: February 2021

Combination therapies constitute a powerful tool for cancer treatment. By combining drugs with different mechanisms of action, the limitations of each individual agent can be overcome, while increasing therapeutic benefit. Here, we propose employing tumor-migrating decidua-derived mesenchymal stromal cells as therapeutic agents combining antiangiogenic therapy and chemotherapy. First, a plasmid encoding the antiangiogenic protein endostatin was transfected into these cells by nucleofection, confirming its expression by ELISA and its biological effect in an ex ovo chick embryo model. Second, doxorubicin-loaded mesoporous silica nanoparticles were introduced into the cells, which would act as vehicles for the drug being released. The effect of the drug was evaluated in a coculture in vitro model with mammary cancer cells. Third, the combination of endostatin transfection and doxorubicin-nanoparticle loading was carried out with the decidua mesenchymal stromal cells. This final cell platform was shown to retain its tumor-migration capacity in vitro, and the combined in vitro therapeutic efficacy was confirmed through a 3D spheroid coculture model using both cancer and endothelial cells. The results presented here show great potential for the development of combination therapies based on genetically-engineered cells that can simultaneously act as cellular vehicles for drug-loaded nanoparticles.
Anti-Tumor Efficiency of Perillylalcohol/β-Cyclodextrin Inclusion Complexes in a Sarcoma S180-Induced Mice Model
Authors: Rezende Allan A. , Santos Rafael S. , Andrade Luciana N. , Amaral Ricardo G. , Pereira Matheus M. , Bani Cristiane , Chen Mo , Priefer Ronny , da Silva Classius F. , de Albuquerque Júnior Ricardo L. C. , Souto Eliana B. , Severino Patrícia
Published: February 2021

The low solubility and high volatility of perillyl alcohol (POH) compromise its bioavailability and potential use as chemotherapeutic drug. In this work, we have evaluated the anticancer activity of POH complexed with β-cyclodextrin (β-CD) using three complexation approaches. Molecular docking suggests the hydrogen-bond between POH and β-cyclodextrin in molar proportion was 1:1. Thermal analysis and Fourier-transform infrared spectroscopy (FTIR) confirmed that the POH was enclosed in the β-CD cavity. Also, there was a significant reduction of particle size thereof, indicating a modification of the β-cyclodextrin crystals. The complexes were tested against human L929 fibroblasts after 24 h of incubation showing no signs of cytotoxicity. Concerning the histopathological results, the treatment with POH/β-CD at a dose of 50 mg/kg promoted approximately 60% inhibition of tumor growth in a sarcoma S180-induced mice model and the reduction of nuclear immunoexpression of the Ki67 antigen compared to the control group. Obtained data suggest a significant reduction of cycling cells and tumor proliferation. Our results confirm that complexation of POH/β-CD not only solves the problem related to the volatility of the monoterpene but also increases its efficiency as an antitumor agent.
Current Perspective on Nasal Delivery Systems for Chronic Rhinosinusitis
Authors: Tai Junhu , Lee Kijeong , Kim Tae Hoon
Published: February 2021

Chronic rhinosinusitis is an upper respiratory disease during which topical drug treatment via the nasal cavity is the most actively utilized therapeutic strategy. In addition to steroids, antibiotics, and antifungal agents, which are widely used in clinical practice, research on novel topical agents to improve the bacterial biofilm or mucociliary clearance remains ongoing. Moreover, owing to the complex structure of the nasal cavity, the effects of nasal drug delivery vary depending on factors related to delivery fluid dynamics, including device, volume, and compounds. In this article, we review methods and compounds that have been applied to chronic rhinosinusitis management and introduce recent advances and future perspectives in nasal drug delivery for upper respiratory diseases.
Fundamental Investigations into Metoprolol Tartrate Deposition on Orodispersible Films by Inkjet Printing for Individualised Drug Dosing
Authors: Kiefer Olga , Fischer Björn , Breitkreutz Jörg
Published: February 2021

Individualised medicine is continuously gaining attention in pharmaceutical research. New concepts and manufacturing technologies are required to realise this therapeutic approach. Off-label drugs used in paediatrics, such as metoprolol tartrate (MPT), are potential candidates for innovations in this context. Orodispersible films (ODFs) have been shown as an accepted alternative dosage form during the last years and inkjet printing is traded as seminal technology of precise deposition of active pharmaceutical ingredients (APIs). The objective of this study was to combine both technologies by developing imprinted ODFs based on hypromellose with therapeutically reasonable MPT single doses of 0.35 to 3.5 mg for paediatric use. After preselection, suitable ink compositions were analysed by confocal Raman microscopy regarding MPT distribution within the imprinted ODFs. Adjusted print settings, speed, print direction and angle, characterised the final ODF surface structure. The present investigations show that uniform dosages with acceptance values between 1 and 6 can be achieved. Nevertheless, changes in calibrated printed quantity due to nozzle aging have a significant effect on the final applied dose. At the lowest investigated quantity, the RSD was ±28% and at the highest, ±9%. This has to be considered for implementation of inkjet printing as a pharmaceutical production tool in the future.
Intracellular Delivery of Active Proteins by Polyphosphazene Polymers
Authors: Qamar Bareera , Solomon Melani , Marin Alexander , Fuerst Thomas R. , Andrianov Alexander K. , Muro Silvia
Published: February 2021

Achieving intracellular delivery of protein therapeutics within cells remains a significant challenge. Although custom formulations are available for some protein therapeutics, the development of non-toxic delivery systems that can incorporate a variety of active protein cargo and maintain their stability, is a topic of great relevance. This study utilized ionic polyphosphazenes (PZ) that can assemble into supramolecular complexes through non-covalent interactions with different types of protein cargo. We tested a PEGylated graft copolymer (PZ-PEG) and a pyrrolidone containing linear derivative (PZ-PYR) for their ability to intracellularly deliver FITC-avidin, a model protein. In endothelial cells, PZ-PYR/protein exhibited both faster internalization and higher uptake levels than PZ-PEG/protein, while in cancer cells both polymers achieved similar uptake levels over time, although the internalization rate was slower for PZ-PYR/protein. Uptake was mediated by endocytosis through multiple mechanisms, PZ-PEG/avidin colocalized more profusely with endo-lysosomes, and PZ-PYR/avidin achieved greater cytosolic delivery. Consequently, a PZ-PYR-delivered anti-F-actin antibody was able to bind to cytosolic actin filaments without needing cell permeabilization. Similarly, a cell-impermeable Bax-BH3 peptide known to induce apoptosis, decreased cell viability when complexed with PZ-PYR, demonstrating endo-lysosomal escape. These biodegradable PZs were non-toxic to cells and represent a promising platform for drug delivery of protein therapeutics.
A New Surface Charge Neutralizing Nano-Adjuvant to Potentiate Polymyxins in Killing Mcr-1 Mediated Drug-Resistant Escherichia coli
Authors: Cho Hyejin , Naskar Atanu , Lee Sohee , Kim Semi , Kim Kwang-Sun
Published: February 2021

Resistance to polymyxins when treating multidrug-resistant (MDR) Gram-negative bacterial infections limit therapeutic options. Here, we report the synthesis of a nickel (Ni) doped Zinc oxide (NZO) combined with black phosphorus (BP) (NZB) nanocomposite and its synergistic action with polymyxin B (PolB) against polymyxin-resistant Escherichia coli harboring mobilized colistin resistance (mcr-1) gene. NZB and PolB combination therapy expressed a specific and strong synergy against Mcr-1 expressing E. coli cells. The underlying mechanism of the synergy is the charge neutralization of the E. coli cell surface by NZB, resulting in a more feasible incorporation of PolB to E. coli. The synergistic concentration of NZB with PolB was proved biocompatible. Thus, the NZB is the first biocompatible nano-adjuvant to polymyxins against polymyxin-resistant E. coli cells, recognizing the physical status of bacteria instead of known adjuvants targeting cellular gene products. Therefore, NZB has the potential to revive polymyxins as leading last-resort antibiotics to combat polymyxin-resistant Gram-negative bacterial infections.
Preparation of Electrospun Small Intestinal Submucosa/Poly(caprolactone-co-Lactide-co-glycolide) Nanofiber Sheet as a Potential Drug Carrier
Authors: Thao Nguyen Thi Thu , Lee Surha , Shin Gi Ru , Kang Youngji , Choi Sangdun , Kim Moon Suk
Published: February 2021

In this work, we chose small intestine submucosa (SIS) as a drug carrier because SIS possesses good biocompatibility, non-immunogenic property and bio-resorbability, and performed electrospinning for preparation of nanofiber sheets (NS). For the preparation of drug-loaded electrospun SIS nanofiber sheets as a drug carrier, we used poly(ε-caprolactone-ran-l-lactide) (PCLA) copolymers to improve the electrospinning performance of SIS. The electrospinning of SIS and PCLA provided the electrospun SIS/PCLA (S/P)-nanofiber sheet (S/P-NS) with adjustable thickness and areas. The electrospun S/P-NS showed different porosities, pore sizes, diameters and tensile strengths depending on the ratios between SIS and PCLA. The electrospun S/P-NS was used as a drug carrier of the dexamethasone (Dex) and silver sulfadiazine (AgS) drug related to anti-inflammation. Dex-loaded S/P-NS and AgS-loaded S/P-NS was successfully fabricated by the electrospinning. In the in vitro and in vivo release, we successfully confirmed the possibility for the sustained release of Dex and AgS from the Dex-S/P-NS and AgS-S/P-NS for three weeks. In addition, the sustained Dex and AgS release suppressed the macrophage infiltration. Collectively, we achieved feasible development of SIS nanofiber sheets for a sustained Dex and AgS delivery system.
Implementation of a Teaching Electronic Medical Record within Didactic Instruction Using a Drug Information Question Assignment
Authors: Wasynczuk Jacqueline , Sheehan Amy H.
Published: February 2021

Background: Pharmacy graduates are expected to be practice-ready to deliver quality patient care, which includes having comprehensive knowledge of health informatics and electronic medical records (EMRs). The purpose of this study was to (1) incorporate an EMR within a pharmacy student assignment, and (2) assess student perceptions of use of the EMR. Methods: Student pharmacists received a patient-specific drug-related question and were required to use an EMR to provide an accurate response. Following completion of the assignment, students were invited to complete a retrospective, pre-post survey instrument to collect their perceptions. Results: Only 28.8% of respondents reported prior experience using an EMR. Student perceptions about use of an EMR within the didactic setting significantly improved from before to after the assignment. Differences were found in respondents who agreed that didactic use of an EMR increased their confidence in obtaining information from an EMR (20.5% to 82.8%) and improved their knowledge of EMR systems (61.4% to 89.3%). Conclusions: Implementation of an EMR within didactic instruction may serve as the first exposure to health informatics for students and positively impacts student perceptions of these tools prior to entry into pharmacy practice.
Integration of Intercultural Learning into an International Advanced Pharmacy Practice Experience in London, England
Authors: Schellhase Ellen , Hasan Ishmum , Hendricks Stephanie , Miller Monica L.
Published: February 2021

As the number of international advanced pharmacy practice experiences (APPEs) continues to grow, this is an opportunity to incorporate intercultural learning (ICL) to further advance student pharmacist training. Purdue University student pharmacists participated in a clinical research focused APPE in London, England. To prepare for this APPE, students completed a one-credit course focused on intercultural learning and travel preparation. The purpose of this report is to describe the implementation and assessment of ICL during this course and international APPE. The course includes interactive ICL activities, reflective assignments, and personalized assessments. During the eight-week APPE, student pharmacists worked on an individualized Intercultural Development Plan®, which includes ICL activities, focused reflection, and check-ins. ICL was assessed using the Intercultural Development Inventory® (IDI®) at the beginning of the course and at least four weeks after APPE completion. Student APPE feedback was also reviewed for evidence of ICL. Twenty-seven students completed the course and APPE from 2018 to 2020. The average IDI developmental orientation (DO) before the course was 91.7, placing students in minimization. The average perceived orientation was 120.9, placing students in acceptance. There were 18 students who completed the post-APPE IDI: 12 students demonstrated growth in the DO (range: 1.5–23.72), and six students experienced a decrease in their DO. Intercultural learning can be implemented and assessed as part of an international APPE.
Monochromatic Conical IR Emission from Decaying KrF Laser Filaments in Xenon as Coherent Stimulated Four-Wave Mixing Process
Authors: Smetanin Igor V. , Shutov Alexey V. , Ustinovskii Nikolay N. , Zvorykin Vladimir D. , Bogatskaya Anna V. , Popov Alexander M.
Published: February 2021

We develop theoretical background for the new nonlinear optical phenomenon of narrowly directed monochromatic IR conical emission which has been recently observed when 248-nm UV filaments propagate in xenon (V. D. Zvorykin, et al., Laser Phys. Lett. 13, 125404 (2016)). We treat it as coherent stimulated four-wave mixing process in which two pump KrF laser photons are converted into the coupled pair of resonance IR(828 nm) and VUV (147 nm) photons through 5p5(2P3/2)6p[1/2]0→5p5(2P3/2)6s[3/2]1o and 5p5(2P3/2)6s[3/2]1o→1S0 transitions. We explore the coherent interaction regime which proceeds at a time scale shorter than transverse relaxation time T2. The momentum and energy conservation laws determine the characteristic angle of conical emission. We find that the threshold of this coherent process is determined by the KrF laser pump pulse area.
Sensitive, Accurate and Rapid Detection of the Northern Root-Knot Nematode, Meloidogyne hapla, Using Recombinase Polymerase Amplification Assays
Authors: Subbotin Sergei A. , Burbridge Julie
Published: February 2021

Rapid and reliable diagnostics of root-knot nematodes are critical for selections of effective control against these agricultural pests. In this study, recombinase polymerase amplification (RPA) assays were developed targeting the IGS rRNA gene of the northern root-knot nematode, Meloidogyne hapla. The RPA assays using TwistAmp® Basic, TwistAmp® exo and TwistAmp® nfo kits (TwistDx, Cambridge, UK) allowed for the detection of M. hapla from crude extracts of females, eggs and juveniles without a DNA extraction step. The results of the RPA assays using real-time fluorescence detection (real-time RPA) in series of crude nematode extracts showed reliable detection after 13 min with a sensitivity of 1/100 of a second-stage juvenile and up to 1/1000 of a female in reaction tubes. The results of the RPA assays using lateral flow dipsticks (LF-RPA) showed reliable detection within 30 min with a sensitivity of 1/10 of a second-stage juvenile and 1/1000 of a female in reaction tubes. The RPA assay developed here is a successful tool for quick, accurate and sensitive diagnostics of M. hapla. The application of the LF-RPA assay has great potential for diagnosing infestation of this species in the lab, field or in areas with a minimal laboratory infrastructure.
Oxidative Stress and Antioxidative Activity in Leaves and Roots of Carrot Plants Induced by Candidatus Phytoplasma Solani
Authors: Mitrovic Petar , Djalovic Ivica , Kiprovski Biljana , Veljović Jovanović Sonja , Trkulja Vojislav , Jelušić Aleksandra , Popović Tatjana
Published: February 2021

The present study examined the effects of Candidatus Phytoplasma solani infection on antioxidative metabolism in leaves and roots of carrot (Daucus carota L.). Disease symptoms appeared at the end of June in the form of the chlorosis on some of the leaves, which became intensely red one week later, while the previously healthy leaves from the same branch becme chlorotic. A few days later, all leaves from the infected leaf branch were intensely red. Infected plants also had slower growth compared to the healthy ones with fewer leaf branches developed. The roots of infected plants were less developed, seared, or gummy with or without brown-colored root hair. The presence of the pathogen was detected by sequencing the 16S rRNA. National Center for Biotechnology Information (NCBI) BLAST analyses of the obtained sequence revealed 100% identity of tested strain with deposited Ca. Phytoplasma solani strains from various countries and hosts, all belonging to the “stolbur” group (16SrXII-A). Identity of 99.74% was found when the tested Serbian strain (MF503627) was compared with the reference stolbur strain STOL11 (AF248959). The oxidative damage of membranes in carrot cells was accompanied by a decrease in the content of photosynthetic pigments. Furthermore, for the determination of specific scavenging properties of the extracts, in vitro antioxidant assay was performed. In phytoplasma-infected carrot leaves, there was a greater reduction in the level of glutathione content (GSH); however; flavonoids and anthocyanidins seem to be responsible for the accompanied increased antioxidative capacity against hydroxyl radical and hydrogen peroxide.
Generation of Chloroplast Molecular Markers to Differentiate Sophora toromiro and Its Hybrids as a First Approach to Its Reintroduction in Rapa Nui (Easter Island)
Authors: Pezoa Ignacio , Villacreses Javier , Rubilar Miguel , Pizarro Carolina , Galleguillos María Jesús , Ejsmentewicz Troy , Fonseca Beatriz , Espejo Jaime , Polanco Víctor , Sánchez Carolina
Published: February 2021

Sophora toromiro is an endemic tree of Rapa Nui with religious and cultural relevance that despite being extinct in the wild, still persists in botanical gardens and private collections around the world. The authenticity of some toromiro trees has been questioned because the similarities among hybrid lines leads to misclassification of the species. The conservation program of toromiro has the objective of its reinsertion into Rapa Nui, but it requires the exact genotyping and certification of the selected plants in order to efficiently reintroduce the species. In this study, we present for the first time the complete chloroplast genome of S. toromiro and four other Sophora specimens, which were sequenced de-novo and assembled after mapping the raw reads to a chloroplast database. The length of the chloroplast genomes ranges from 154,239 to 154,473 bp. A total of 130–143 simple sequence repeats (SSR) loci and 577 single nucleotide polymorphisms (SNPs) were identified.
Simultaneous Biochemical and Physiological Responses of the Roots and Leaves of Pancratium maritimum (Amaryllidaceae) to Mild Salt Stress
Authors: Carfagna Simona , Salbitani Giovanna , Innangi Michele , Menale Bruno , De Castro Olga , Di Martino Catello , Crawford Thomas W.
Published: February 2021

Pancratium maritimum (Amaryllidaceae) is a bulbous geophyte growing on coastal sands. In this study, we investigated changes in concentrations of metabolites in the root and leaf tissue of P. maritimum in response to mild salt stress. Changes in concentrations of osmolytes, glutathione, sodium, mineral nutrients, enzymes, and other compounds in the leaves and roots were measured at 0, 3, and 10 days during a 10-day exposure to two levels of mild salt stress, 50 mM NaCl or 100 mM NaCl in sandy soil from where the plants were collected in dunes near Cuma, Italy. Sodium accumulated in the roots, and relatively little was translocated to the leaves. At both concentrations of NaCl, higher values of the concentrations of oxidized glutathione disulfide (GSSG), compared to reduced glutathione (GSH), in roots and leaves were associated with salt tolerance. The concentration of proline increased more in the leaves than in the roots, and glycine betaine increased in both roots and leaves. Differences in the accumulation of organic osmolytes and electron donors synthesized in both leaves and roots demonstrate that osmoregulatory and electrical responses occur in these organs of P. maritimum under mild salt stress.
Resistance against Orobanche crenata in Bitter Vetch (Vicia ervilia) Germplasm Based on Reduced Induction of Orobanche Germination
Authors: González-Verdejo Clara Isabel , Fernández-Aparicio Mónica , Córdoba Eva María , López-Ráez Juan Antonio , Nadal Salvador
Published: February 2021

Bitter vetch (Vicia ervilia (L.) Willd.) is a legume well adapted to cultivation in marginal areas, being an important source of protein for animal feed in low input cropping systems. Surprisingly, it is an underutilized crop as it could be a good alternative to increase the sustainability of extensive rainfed cropping systems. In Mediterranean rainfed cropping systems, the productivity of bitter vetch is severely reduced by the parasitic weed species Orobanche crenata (Forsk). To date, few resistant bitter vetch genotypes have been identified. O. crenata infection process initiates with the recognition of germination factors exuded by roots of susceptible hosts. In this work, the interaction of a collection of bitter vetch accessions and O. crenata has been analyzed in order to discover accessions with low germination induction activity. Through a combination of field and rhizotron experiments, two bitter vetch accessions were selected showing low germination-induction activity, which resulted in less infection. In addition, in vitro germination assays revealed that the low germination activity was due to low exudation of germination factors and not due to the exudation of germination inhibitors. The selected low germination-inducers genotypes could be the basis for a new breeding program generating locally adapted alternatives with resistance to O. crenata.
Water Vapor Sorption and Diffusivity in Bio-Based Poly(ethylene vanillate)—PEV
Authors: Giacobazzi Greta , Gioia Claudio , Vannini Micaela , Marchese Paola , Guillard Valérie , Angellier-Coussy Hélène , Celli Annamaria
Published: February 2021

The dynamic and equilibrium water vapor sorption properties of amorphous and highly crystalline poly(ethylene vanillate) (PEV) films were determined via gravimetric analysis, at 20 °C, over a wide range of relative humidity (0–95% RH). At low RH%, the dynamic of the sorption process obeys Fick’s law while at higher relative humidity it is characterized by a drift ascribable to non-Fickian relaxations. The non-Fickian relaxations, which are responsible for the incorporation of additional water, are correlated with the upturn of the sorption isotherms and simultaneously the hysteresis recorded between sorption and desorption cycles. The sorption isotherms of amorphous and highly crystalline PEV are arranged in the same concentration range of that of PET proving the similarity of the two polyesters. Water diffusion coefficients, whose determination from individual kinetic sorption/desorption curves required treatment with the Barens–Hopfenberg model, were demonstrated to be ≈10× higher for amorphous PEV compared to amorphous PET. Such a difference originates from the enhanced segmental flexibility of PEV chains.
Hardness Modulated Thermoplastic Poly(ether ester) Elastomers for the Automobile Weather-Strip Application
Authors: Jeon Su Hyeon , Jeong Jae Eon , Kim Seongkyun , Jeon Sungwan , Choung Jin Woo , Kim Il
Published: February 2021

As a means of developing new material for automobile weather-stripping and seal parts replacing the conventional ethylene propylene diene monomer rubber/polypropylene vulcanizate, a series of poly(ether ester) elastomers are synthesized. The hardness is modulated by controlling chain extender composition after fixing the hard segment to soft segment ratio. Targeted hardness is achieved by partly substituting conventional chain extender 1,4-butandiol for soybean oil-originated fatty acid amide diol that bears a long chain branch. The crystallinity and phase separation behavior resultant elastomer are also tunable simply by modulating chain extender composition and hard to soft segment ratio.
Curdlan–Chitosan Electrospun Fibers as Potential Scaffolds for Bone Regeneration
Authors: Toullec Clément , Le Bideau Jean , Geoffroy Valerie , Halgand Boris , Buchtova Nela , Molina-Peña Rodolfo , Garcion Emmanuel , Avril Sylvie , Sindji Laurence , Dube Admire , Boury Frank , Jérôme Christine
Published: February 2021

Polysaccharides have received a lot of attention in biomedical research for their high potential as scaffolds owing to their unique biological properties. Fibrillar scaffolds made of chitosan demonstrated high promise in tissue engineering, especially for skin. As far as bone regeneration is concerned, curdlan (1,3-β-glucan) is particularly interesting as it enhances bone growth by helping mesenchymal stem cell adhesion, by favoring their differentiation into osteoblasts and by limiting the osteoclastic activity. Therefore, we aim to combine both chitosan and curdlan polysaccharides in a new scaffold for bone regeneration. For that purpose, curdlan was electrospun as a blend with chitosan into a fibrillar scaffold. We show that this novel scaffold is biodegradable (8% at two weeks), exhibits a good swelling behavior (350%) and is non-cytotoxic in vitro. In addition, the benefit of incorporating curdlan in the scaffold was demonstrated in a scratch assay that evidences the ability of curdlan to express its immunomodulatory properties by enhancing cell migration. Thus, these innovative electrospun curdlan–chitosan scaffolds show great potential for bone tissue engineering.
2D Correlation Spectroscopy (2DCoS) Analysis of Temperature-Dependent FTIR-ATR Spectra in Branched Polyethyleneimine/TEMPO-Oxidized Cellulose Nano-Fiber Xerogels
Authors: Paladini Giuseppe , Venuti Valentina , Crupi Vincenza , Majolino Domenico , Fiorati Andrea , Punta Carlo
Published: February 2021

Fourier transform infrared spectroscopy in attenuated total reflectance geometry (FTIR-ATR), combined with a 2D correlation analysis, was here employed to investigate temperature-induced spectral changes occurring in a particular type of novel cellulosic-based nano-material prepared using 2,2,6,6-tetramethyl-piperidine-1-oxyl (TEMPO) oxidized and ultra-sonicated cellulose nano-fibers (TOUS-CNFs) as three-dimensional scaffolds, and branched polyethyleneimine (bPEI) as cross-linking agent. The aim was to highlight the complex sequential events involving the different functional groups of the polymeric network, as well as to gain insight into the interplay between the amount of bPEI and the resulting sponge-like material, upon increasing temperature. In this framework, synchronous and asynchronous 2D spectra were computed and analyzed in three wavenumber regions (900–1200 cm−1, 1500–1700 cm−1 and 2680–3780 cm−1), where specific vibrational modes of the cellulosic structure fall, and over a T-range between 250 K and 340 K. A step-by-step evolution of the different arrangements of the polymer functional groups was proposed, with particular regard to how the cooperativity degree of inter- and intramolecular hydrogen bonds (HBs) changes upon heating. Information acquired can be useful, in principle, in order to develop a next-generation, T-sensitive novel material to be used for water remediation applications or for drug-delivery nano-vectors.
Determination of the Operational Parameters for the Manufacturing of Spherical PVP Particles via Electrospray
Authors: Narváez-Muñoz Christian , Ryzhakov Pavel , Pons-Prats Jordi
Published: February 2021

This work aims at bridging experimental and numerical approaches to determine the optimal operating parameters for the fabrication of well-shaped polyvinylpyrrolidone (PVP) particles via electrohydrodynamic atomization. Particular emphasis is given to the role of the PVP solution viscosity. Solutions of PVP at various concentrations dissolved in Dimethylformamide (DMF) were prepared and analyzed. Numerical simulation using a coupled electro-CFD model was used to determine the ranges of experimental flow rate and the voltage, ensuring that well-shaped spherical particles are produced. It was deduced that the optimal combination of the parameters (flow rate, voltage, and polymer concentration) can be well approximated by a scaling law. The established relationship allowed determination of a stability island that guarantees that the given polymer solution will form spherical particles. Analyzing morphology and sizes of the particles manufactured in the optimal parameters range, we show, among others, that the size of the PVP particles can be predicted as a function of the flow rate by a power scaling relationship.
Biomedical Applications of Bacterial Exopolysaccharides: A Review
Authors: Mohd Nadzir Masrina , Nurhayati Retno Wahyu , Idris Farhana Nazira , Nguyen Minh Hong
Published: February 2021

Bacterial exopolysaccharides (EPSs) are an essential group of compounds secreted by bacteria. These versatile EPSs are utilized individually or in combination with different materials for a broad range of biomedical field functions. The various applications can be explained by the vast number of derivatives with useful properties that can be controlled. This review offers insight on the current research trend of nine commonly used EPSs, their biosynthesis pathways, their characteristics, and the biomedical applications of these relevant bioproducts.
Solution Blowing Spinning Technology towards Green Development of Urea Sensor Nanofibers Immobilized with Hydrazone Probe
Authors: El-Newehy Mohamed H. , El-Hamshary Hany , Salem Waheed M.
Published: February 2021

Cellulose has been one of the most widespread materials due to its renewability, excellent mechanical properties, biodegradability, high absorption ability, biocompatibility and cheapness. Novel, simple and green colorimetric fibrous film sensor was developed by immobilization of urease enzyme (U) and tricyanofuran hydrazone (TCFH) molecular probe onto cellulose nanofibers (CNF). Cellulose acetate nanofibers (CANF) were firstly prepared from cellulose acetate using the simple, green and low cost solution blowing spinning technology. The produced CANF was exposed to deacetylation to introduce CNF, which was then treated with a mixture of TCFH and urease enzyme to introduce CNF-TCFH-U nanofibrous biosensor. CNF were reinforced with tricyanofuran hyrazone molecular probe and urease enzyme was encapsulated into calcium alginate biopolymer to establish a biocomposite film. This CNF-TCFH-U naked-eye sensor can be applied as a disposable urea detector. CNF demonstrated a large surface area and was utilized as a carrier for TCFH, which is the spectroscopic probe and urease is a catalyst. The biochromic CNF-TCFH-U nanofibrous biosensor responds to an aqueous medium of urea via a visible color signal changing from off-white to dark pink. The morphology of the generated CNF and CNF-TCFH-U nanofibrous films were characterized by different analytical tools, including energy-dispersive X-ray patterns (EDX), polarizing optical microscope (POM), Fourier-transform infrared spectroscopy (FT-IR) and scanning electron microscope (SEM). SEM images of CNF-TCFH-U nanofibers demonstrated diameters between 800 nm and 2.5 μm forming a nonwoven fabric with a homogeneous distribution of TCFH/urease-containing calcium alginate nanoparticles on the surface of CNF. The morphology of the cross-linked calcium alginate nanoparticles was also explored using transmission electron microscopy (TEM) to indicate an average diameter of 56–66 nm. The photophysical performance of the prepared CNF-TCFH-U was also studied by CIE Lab coloration parameters. The nanofibrous film biosensor displayed a relatively rapid response time (5–10 min) and a limit of detection as low as 200 ppm and as high as 1400 ppm. Tricyanofuran hydrazone is a pH-responsive disperse dye comprising a hydrazone detection group. Determination of urea occurs through a proton transfer from the hydrazone group to the generated ammonia from the reaction of urea with urease.