bluuebunny/crossref_metadata_2025 huggingface dataset:
Synthesis of Block Copolymer Brush by RAFT and Click Chemistry and Its Self-Assembly as a Thin Film
Authors: Thankappan Hajeeth , Semsarilar Mona , Li Suming , Chang Yung , Bouyer Denis , Quemener Damien
Published: October 2020

A well-defined block copolymer brush poly(glycidyl methacrylate)-graft-(poly(methyl methacrylate)-block- poly(oligo(ethylene glycol) methyl ether methacrylate)) (PGMA-g-(PMMA-b-POEGMA)) is synthesized via grafting from an approach based on a combination of click chemistry and reversible addition-fragmentation chain transfer (RAFT) polymerization. The resulting block copolymer brushes were characterized by 1H-NMR and size exclusion chromatography (SEC). The self-assembly of the block copolymer brush was then investigated under selective solvent conditions in three systems: THF/water, THF/CH3OH, and DMSO/CHCl3. PGMA-g-(PMMA-b-POEGMA) was found to self-assemble into spherical micelle structures as analyzed by transmission electron microscopy (TEM) and dynamic light scattering (DLS). The average size of the particles was much smaller in THF/CH3OH and DMSO/CHCl3 as compared with the THF/water system. Thin film of block copolymer brushes with tunable surface properties was then prepared by the spin-coating technique. The thickness of the thin film was confirmed by scanning electron microscopy (SEM). Atom force microscopy (AFM) analysis revealed a spherical morphology when the block copolymer brush was treated with poor solvents for the backbone and hydrophobic side chains. The contact angle measurements were used to confirm the surface rearrangements of the block copolymer brushes.
Ecotoxicity Assessment of Graphene Oxide by Daphnia magna through a Multimarker Approach from the Molecular to the Physiological Level including Behavioral Changes
Authors: Fekete-Kertész Ildikó , László Krisztina , Terebesi Csilla , Gyarmati Benjámin Sándor , Farah Shereen , Márton Rita , Molnár Mónika
Published: October 2020

The extensive use of engineered nanomaterials, such as graphene oxide (GO), is stimulating research about its potential environmental impacts on the aquatic ecosystem. This study is aimed to comprehensively assess the acute toxicity of a well-characterized GO suspension to Daphnia magna. Conventional ecotoxicological endpoints (lethality, immobilization) and more sensitive, sublethal endpoints (heartbeat rate, feeding activity, and reactive oxygen species (ROS)) production were used. The possible normalization of the heartbeat rate and feeding activity in clean test medium was also investigated. The fate, time-dependent, and concentration-dependent aggregation behaviour of GO was followed by dynamic light scattering, UV-Vis spectroscopy, and zeta potential measurement methods. The EC20 value for immobilization was 50 mg/L, while, for physiological and behavioural endpoints, it ranged from 8.1 mg/L (feeding activity) to 14.8 mg/L (immobilization). The most sensitive endpoint was the ROS production with EC20 = 4.78 mg/L. 24-h recovery experiments revealed that feeding activity was restored only up to a certain level at higher concentrations, indicating that the potential environmental health effects of GO cannot be neglected. Alterations of normal physiology (heart rate) and feeding activity may be associated with increased risk of predation and reproductive decline, highlighting that GO may have impacts on population and food web dynamics in aquatic ecosystems.
In Situ Ultraviolet Polymerization Using Upconversion Nanoparticles: Nanocomposite Structures Patterned by Near Infrared Light
Authors: Jee Hongsub , Chen Guanying , Prasad Paras N. , Ohulchanskyy Tymish Y. , Lee Jaehyeong
Published: October 2020

In this paper, we report an approach to polymerization of a nanocomposite containing UV-polymerizable organic material and inorganic, NaYbF4:Tm3+ core-based nanoparticles (NPs), which are optimized for upconversion of near infrared (NIR) to ultraviolet (UV) and blue light. Our approach is compatible with numerous existing UV-polymerizable compositions and the NaYF4: Yb, Tm3+ core-based NPs are much more stable against harsh conditions than NIR organic photo-initiators proposed earlier. The use of a core-shell design for the NPs can provide a suitable method for binding with organic constituents of the nanocomposite, while maintaining efficient NIR-to-UV/blue conversion in the NaYbF4 core. The prepared photopolymerized transparent polymer nanocomposites display upconversion photoluminescence in UV, visible and NIR ranges. We also demonstrate a successful fabrication of polymerized nanocomposite structure with millimeter/submillimeter size uniformly patterned by 980 nm irradiation of inexpensive laser diode through a photomask.
PANS/PANDAS: Clinical Experience in IVIG Treatment and State of the Art in Rehabilitation Approaches
Authors: Pavone Piero , Falsaperla Raffaele , Cacciaguerra Giovanni , Sapuppo Annamaria , Chiaramonte Rita , Lubrano Riccardo , Messina Giulia , Sciuto Sarah , Sabino Luca , Cocuzza Salvatore , Maniaci Antonino , Fontana Alessandra , Marino Lidia , Oliva Claudia , Pappalardo Maria Grazia , Vecchio Michele
Published: October 2020

Pediatric acute-onset neuropsychiatric syndrome (PANS) is a condition characterized by the abrupt, dramatic onset of obsessive–compulsive disorder (OCD) or eating restriction accompanied by equally abrupt and severe comorbid neuropsychiatric symptoms. PANDAS (pediatric autoimmune neuropsychiatric disorder associated with streptococcal infection) is a heterogeneous syndrome identified as post-Streptococcus pyogenes infection (β-hemolytic Streptococcus group A) complications regarding the central nervous system with specific involvement of neuropsychiatric and behavioral skills. In the first part of our study, we share our experience in the treatment of a group of extreme-grade (according to CY-BOCS severity scale) symptomatic patients with intravenous immunoglobulin (IVIG), following the most recent studies regarding the dosage of the drug. Our contribution is to share our experience made on a sample of 55 patients all in the highest level of a severity grade. In the second part of our study, we also analyze the literature on PANS/PANDAS rehabilitation therapy, since in the literature there is no discussion of union and comparison on this method. Objective: This study aims to evaluate the clinical features of the patients observed from different Italian cohorts, with the attempt at evaluating clinical response to IVIG treatment in children with an extreme severity grade of PANS/PANDAS disease. Furthermore, after having analyzed the literature, we propose rehabilitation therapy as an added value to the pharmacological treatment. Materials and Methods: A total of 55 patients with a diagnosis of PANS/PANDAS, who belonged to an extreme grade of disease, were enrolled. All patients were administered with IVIG treatment at 2 g/kg per day for two consecutive days. Results: From our study, a noticeable improvement (until complete remission) of symptoms was evident for at least one year in 47 out of 55 (85%) observed children, while 11 out of these 43 (25%) showed an evident symptoms remission in a single attempt and the remaining 32 (75%) required a second administration to notice a lasting symptomatic improvement.
The Causal Effects of Blood Iron and Copper on Lipid Metabolism Diseases: Evidence from Phenome-Wide Mendelian Randomization Study
Authors: Zhou Jingqi , Liu Chang , Francis Michael , Sun Yitang , Ryu Moon-Suhn , Grider Arthur , Ye Kaixiong
Published: October 2020

Blood levels of iron and copper, even within their normal ranges, have been associated with a wide range of clinical outcomes. The available epidemiological evidence for these associations is often inconsistent and suffers from confounding and reverse causation. This study aims to examine the causal clinical effects of blood iron and copper with Mendelian randomization (MR) analyses. Genetic instruments for the blood levels of iron and copper were curated from existing genome-wide association studies. Candidate clinical outcomes were identified based on a phenome-wide association study (PheWAS) between these genetic instruments and a wide range of phenotypes in 310,999 unrelated individuals of European ancestry from the UK Biobank. All signals passing stringent correction for multiple testing were followed by MR analyses, with replication in independent data sources where possible. We found that genetically predicted higher blood levels of iron and copper are both associated with lower risks of iron deficiency anemia (odds ratio (OR) = 0.75, 95% confidence interval (CI): 0.67–0.85, p = 1.90 × 10−6 for iron; OR = 0.88, 95% CI: 0.78–0.98, p = 0.032 for copper), lipid metabolism disorders, and its two subcategories, hyperlipidemia (OR = 0.90, 95% CI: 0.85–0.96, p = 6.44 × 10−4; OR = 0.92, 95% CI: 0.87–0.98, p = 5.51 × 10−3) and hypercholesterolemia (OR = 0.90, 95% CI: 0.84–0.95, p = 5.34 × 10−4; OR = 0.93, 95% CI: 0.89–0.99, p = 0.022). Consistently, they are also associated with lower blood levels of total cholesterol and low-density lipoprotein cholesterol. Multiple sensitivity tests were applied to assess the presence of pleiotropy and the robustness of causal estimates. Regardless of the approaches, consistent evidence was obtained. Moreover, the unique clinical effects of each blood mineral were identified. Notably, genetically predicated higher blood iron is associated with an enhanced risk of varicose veins (OR = 1.28, 95% CI: 1.15–1.42, p = 4.34 × 10−6), while blood copper is positively associated with the risk of osteoarthrosis (OR = 1.07, 95% CI: 1.02–1.13, p = 0.010). Sex-stratified MR analysis further revealed some degree of sex differences in their clinical effects. Our comparative PheWAS-MR study of iron and copper comprehensively characterized their shared and unique clinical effects, highlighting their potential causal roles in hyperlipidemia and hypercholesterolemia. Given the modifiable nature of blood mineral status and the potential for clinical intervention, these findings warrant further investigation.
Impact of Clinical Markers of Nutritional Status and Feeding Jejunostomy Use on Outcomes in Esophageal Cancer Patients Undergoing Neoadjuvant Chemoradiotherapy
Authors: Jain Rishi , Shaikh Talha , Yee Jia-Llon , Au Cherry , Denlinger Crystal S. , Handorf Elizabeth , Meyer Joshua E. , Dotan Efrat
Published: October 2020

Background: Patients with esophageal cancer (EC) have high rates of malnutrition due to tumor location and treatment-related toxicity. Various strategies are used to improve nutritional status in patients with EC including oral and enteral support. Methods: We conducted a retrospective analysis to determine the impact of malnutrition and prophylactic feeding jejunostomy tube (FJT) placement on toxicity and outcomes in patients with localized EC who were treated with neoadjuvant chemoradiation therapy (nCRT) followed by esophagectomy. Results: We identified 125 patients who were treated with nCRT between 2002 and 2014. Weight loss and hypoalbuminemia occurred frequently during nCRT and were associated with multiple adverse toxicity outcomes including hematologic toxicity, nonhematologic toxicity, grade ≥3 toxicity, and hospitalizations. After adjusting for relevant covariates including the specific nCRT chemotherapy regimen received and the onset of toxicity, there were no significant associations between hypoalbuminemia, weight loss, or FJT placement and relapse-free survival (RFS) or overall survival (OS). FJT placement was associated with less weight loss during nCRT (p = 0.003) but was not associated with reduced toxicity or improved survival. Conclusions: Weight and albumin loss during nCRT for EC are important factors relating to treatment toxicity but not RFS or OS. While pretreatment FJT placement may reduce weight loss, it may not impact treatment tolerance or survival.
Curcumin: Could This Compound Be Useful in Pregnancy and Pregnancy-Related Complications?
Authors: Filardi Tiziana , Varì Rosaria , Ferretti Elisabetta , Zicari Alessandra , Morano Susanna , Santangelo Carmela
Published: October 2020

Curcumin, the main polyphenol contained in turmeric root (Curcuma longa), has played a significant role in medicine for centuries. The growing interest in plant-derived substances has led to increased consumption of them also in pregnancy. The pleiotropic and multi-targeting actions of curcumin have made it very attractive as a health-promoting compound. In spite of the beneficial effects observed in various chronic diseases in humans, limited and fragmentary information is currently available about curcumin’s effects on pregnancy and pregnancy-related complications. It is known that immune-metabolic alterations occurring during pregnancy have consequences on both maternal and fetal tissues, leading to short- and long-term complications. The reported anti-inflammatory, antioxidant, antitoxicant, neuroprotective, immunomodulatory, antiapoptotic, antiangiogenic, anti-hypertensive, and antidiabetic properties of curcumin appear to be encouraging, not only for the management of pregnancy-related disorders, including gestational diabetes mellitus (GDM), preeclampsia (PE), depression, preterm birth, and fetal growth disorders but also to contrast damage induced by natural and chemical toxic agents. The current review summarizes the latest data, mostly obtained from animal models and in vitro studies, on the impact of curcumin on the molecular mechanisms involved in pregnancy pathophysiology, with the aim to shed light on the possible beneficial and/or adverse effects of curcumin on pregnancy outcomes.
Play with Your Food and Cook It! Tactile Play with Fish as a Way of Promoting Acceptance of Fish in 11- to 13-Year-Old Children in a School Setting—A Qualitative Study
Authors: Højer Rikke , Wistoft Karen , Frøst Michael Bom
Published: October 2020

Despite a tradition of consuming fish in Denmark and despite the health benefits of eating fish, Danish children consume only one-third of the officially recommended amount of fish. The objective of this study was to explore an experiential and sensory-based exercise in a school setting with focus on tactile play and cooking as a way of promoting 11- to 13-year-old children’s acceptance of fish. The design was a qualitative exploratory multiple-case design using participant observation in a school setting. Six classes were recruited from the Eastern part of Denmark (n = 132). Based on an exercise with cooking fish and gyotaku (fish print), four meta-themes were identified by applying applied thematic analysis: rejection, acceptance, craftsmanship, and interaction. Rejection and acceptance appeared along a rejection–acceptance continuum related to how the fish was categorised (animal, non-animal, food) in different phases of the experiment. Rejection was promoted by mucus, smell, animalness, and texture, whereas helping each other, tactile play, and craftsmanship promoted acceptance. In conclusion, this study found that tactile play combined with cooking could be a way of promoting acceptance of fish. The findings also support a school setting as a potential gateway in promoting healthy food behaviour.
Nurse Practitioner: Is It Time to Have a Role in Saudi Arabia?
Authors: Almutairi Hessa , Alharbi Kholoud , Alotheimin Hana , Gassas Roaa , Alghamdi Musaad , Alamri Ayman , Alsufyani Abdulaziz , Bashatah Adel
Published: October 2020

Low recruitment of Saudi nationals into the nursing profession, coupled with a growing population, has led to a severe nursing shortage in Saudi Arabia, particularly of nurses with advanced qualifications in clinical nursing. While the role of nurse practitioner has been successfully integrated into the healthcare systems of the U.S., Canada, the UK and Australia for decades, the advanced practice registered nurse (APRN), which includes nurse practitioners and clinical nursing specialists, is still not being implemented effectively in Saudi Arabia due to a variety of regulatory, institutional and cultural barriers. The author looks at some of those barriers and offers recommendations of how they might be overcome. Given that in many parts of the world, nurse practitioners are considered an essential component to meeting healthcare demands, the author considers the question of whether APRNs can find a role in Saudi Arabia’s healthcare system.
Introduction to a New MDPI Open Access Journal: Organics
Authors: Dehaen Wim , Szostak Michal , Xu Huaping
Published: October 2020

Organic chemistry is near to entering its third century, if one dates the Wöhler experiment (1828) as the starting point [...]
The Role of Cathepsins in the Growth of Primary and Secondary Neoplasia in the Bone
Authors: Fasanya Henrietta O. , Siemann Dietmar W.
Published: October 2020

The upregulation of proteolytic enzymes has been demonstrated to promote primary tumor development and metastatic bone cancer. The secreted proteases increase tumor growth and angiogenesis, and potentiate neoplastic cell dissemination. This article reviews the role and mechanisms of cathepsins in normal physiology, cancer, bone remodeling, and the tumor–bone interface, with a specific focus on cathepsins B, D, H, G, L, and K. In this review, we highlight the role of cathepsins in primary bone cancer (i.e., osteosarcoma (OS)), as well as metastatic breast (BCa) and prostate (PCa) cancer. In addition, we discuss the clinical utility and therapeutic potential of cathepsin-targeted treatments in primary and secondary bone cancers.
Onychomycosis in Northwestern Greece over a 7-Year Period
Authors: Sakkas Hercules , Kittas Christos , Kapnisi Georgia , Priavali Efthalia , Kallinteri Amalia , Bassukas Ioannis D. , Gartzonika Konstantina
Published: October 2020

Onychomycosis is considered as one of the major public health problems with a global distribution associated with geographic, demographic and environmental factors, underlying comorbidities and immunodeficiency disorders. This study was conducted to investigate the etiological agents of onychomycosis, in Northwestern Greece during a 7-year period. The study population included 1095 outpatients with clinically suspected onychomycosis that presented to the University Hospital of Ioannina, NW Greece (2011–2017). Samples were examined for causative fungi, and mycological identification was established using standard mycological methods. Demographic data of each patient, comorbidities, localization of infection and history of previous fungal infection were collected. Onychomycosis was diagnosed in 317 of the 1095 suspected cases (28.9%) and the most frequently isolated pathogens were yeasts (50.8%) followed by dermatophytes (36.9%) and non-dermatophyte molds (NDMs) (12.3%). Dermatophytes were mostly involved in toenail onychomycosis (90.6%) and more commonly affected males than females (57.3% vs. 42.7%), while the predominantly isolated pathogen was Τrichophyton rubrum (74.4%) followed by Τrichophyton interdigitale (21.4%). Candida albicans was the most prevalent isolated yeast (82%), whereas among the cases with onychomycosis due to NDMs, Aspergillus spp. were isolated as the principal species (59%). Continuous monitoring should be performed in order to identify possible trends and shifts in species isolation rates and to evaluate the impact of onychomycosis among the general population and high-risk groups.
Homotype-Targeted Biogenic Nanoparticles to Kill Multidrug-Resistant Cancer Cells
Authors: Shair Mohammad Imran , Chaurasiya Birendra , Yang Xuan , Lin Chuchu , Rong Hehui , He Wei
Published: October 2020

“Off-targeting” and receptor density expressed at the target sites always compromise the efficacy of the nanoparticle-based drug delivery systems. In this study, we isolated different cell membranes and constructed cell membrane-cloaked biogenic nanoparticles for co-delivery of antitumor paclitaxel (PTX) and multidrug resistance (MDR)-modulator disulfiram (DSF). Consequently, MDR cancer cell membrane (A549/T)-coated hybrid nanoparticles (A549/T CM-HNPs) selectively recognized the source cells and increased the uptake by ninefold via the homotypic binding mechanism. Moreover, the A549/T CM-HNPs sensitized MDR cells to PTX by suppressing P-glycoprotein (P-gp) activity by 3.2-fold and induced effective apoptosis (70%) in homologous A549/T cells. Cell-membrane coating based on the “homotypic binding” is promising in terms of promoting the accumulation of chemotherapeutics in MDR cells and killing them.
Gellan Gum Promotes the Differentiation of Enterocytes from Human Induced Pluripotent Stem Cells
Authors: Qiu Shimeng , Kabeya Tomoki , Ogawa Isamu , Anno Shiho , Hayashi Hisato , Kanaki Tatsuro , Hashita Tadahiro , Iwao Takahiro , Matsunaga Tamihide
Published: October 2020

The evaluation of drug pharmacokinetics in the small intestine is critical for developing orally administered drugs. Caucasian colon adenocarcinoma (Caco-2) cells are employed to evaluate drug absorption in preclinical trials of drug development. However, the pharmacokinetic characteristics of Caco-2 cells are different from those of the normal human small intestine. Besides this, it is almost impossible to obtain primary human intestinal epithelial cells of the same batch. Therefore, human iPS cell-derived enterocytes (hiPSEs) with pharmacokinetic functions similar to human intestinal epithelial cells are expected to be useful for the evaluation of drug absorption. Previous studies have been limited to the use of cytokines and small molecules to generate hiPSEs. Dietary fibers play a critical role in maintaining intestinal physiology. We used gellan gum (GG), a soluble dietary fiber, to optimize hiPSE differentiation. hiPSEs cocultured with GG had significantly higher expression of small intestine- and pharmacokinetics-related genes and proteins. The activities of drug-metabolizing enzymes, such as cytochrome P450 2C19, and peptide transporter 1 were significantly increased in the GG treatment group compared to the control group. At the end point of differentiation, the percentage of senescent cells increased. Therefore, GG could improve the differentiation efficiency of human iPS cells to enterocytes and increase intestinal maturation by extending the life span of hiPSEs.
Organic Salts Based on Isoniazid Drug: Synthesis, Bioavailability and Cytotoxicity Studies
Authors: Santos Filipa , Branco Luís C. , Duarte Ana Rita C.
Published: October 2020

Tuberculosis is one of the ten causes of morbidity and mortality worldwide caused by Mycobacterium tuberculosis complex. Some of the anti-tuberculosis drugs used in clinic studies, despite being effective for the treatment of tuberculosis, present serious adverse effects as well as poor bioavailability, stability, and drug-resistance problems. Thus, it is important to develop approaches that could provide shorter drug regimens, preventing drug resistance, toxicity of the antibiotics, and improve their bioavailability. Herein, we reported the use of organic salts based on the isoniazid drug, which can act as an organic cation combined with suitable organic anions such as alkylsulfonate-based (mesylate, R or S-Camphorsulfonate), carboxylate-based (glycolate, vanylate) and sacharinate. The synthesis, characterization, and cytotoxicity studies comparing with the original isoniazid drug have been performed. The possibility to explore dicationic salts seems promising in order to improve original bioavailability, and promote the elimination of polymorphic forms as well as higher stability, which are relevant characteristics that the pharmaceutical industry pursues.
Effective Dispensing Methods for Loading Drugs Only to the Tip of DNA Microneedles
Authors: Bok Moonjeong , Zhao Zhi-Jun , Hwang Soon Hyoung , Kang Hyeok-Joong , Jeon Sohee , Ko Jiwoo , Jeong Jiwon , Song Young Seok , Lim Eunju , Jeong Jun-Ho
Published: October 2020

Here, we propose a novel and simple method to efficiently capture the diffusion of fluorescein isothiocyanate (FITC)-dextran from a biocompatible substance and load the drug only to the tip of DNA microneedles. A dispensing and suction method was chosen to fabricate the designed microneedles with efficient amounts of FITC as the drug model. Importantly, the vacuum process, which could influence the capturing of FITC diffusion from the tip, was evaluated during the manufacturing process. In addition, the simulations were consistent with the experimental results and showed apparent diffusion. Moreover, dextrans of different molecular weights labeled with FITC were chosen to fabricate the tip of microneedles for demonstrating their applicability. Finally, a micro-jetting system with a micro-nozzle (diameter: 80 μm) was developed to achieve the accurate and rapid loading of small amounts of FITC using the anti-diffusion and micro-jetting methods. Our method not only uses a simple and fast manufacturing process, but also fabricates the tips of microneedles more efficiently with FITC compared with the existing methods. We believe that the proposed method is essential for the clinical applications of the microneedle drug delivery platform.
Therapeutic Interventions into Innate Immune Diseases by Means of Aptamers
Authors: Yasmeen Farzana , Seo Hana , Javaid Nasir , Kim Moon Suk , Choi Sangdun
Published: October 2020

The immune system plays a crucial role in the body’s defense system against various pathogens, such as bacteria, viruses, and parasites, as well as recognizes non-self- and self-molecules. The innate immune system is composed of special receptors known as pattern recognition receptors, which play a crucial role in the identification of pathogen-associated molecular patterns from diverse microorganisms. Any disequilibrium in the activation of a particular pattern recognition receptor leads to various inflammatory, autoimmune, or immunodeficiency diseases. Aptamers are short single-stranded deoxyribonucleic acid or ribonucleic acid molecules, also termed “chemical antibodies,” which have tremendous specificity and affinity for their target molecules. Their features, such as stability, low immunogenicity, ease of manufacturing, and facile screening against a target, make them preferable as therapeutics. Immune-system–targeting aptamers have a great potential as a targeted therapeutic strategy against immune diseases. This review summarizes components of the innate immune system, aptamer production, pharmacokinetic characteristics of aptamers, and aptamers related to innate-immune-system diseases.
The Encapsulation of Febuxostat into Emulsomes Strongly Enhances the Cytotoxic Potential of the Drug on HCT 116 Colon Cancer Cells
Authors: Fahmy Usama A. , Aldawsari Hibah M. , Badr-Eldin Shaimaa M. , Ahmed Osama A. A. , Alhakamy Nabil A. , Alsulimani Helal H. , Caraci Filippo , Caruso Giuseppe
Published: October 2020

Febuxostat (FBX) is a drug able to inhibit xanthine oxidase and reduce uric acid production commonly used for the treatment of hyperuricemia in subjects suffering from gout. Several studies have also been directed at its use as anti-cancer drug during the last years, opening a window for its off-label use. In the present study, an optimized formulation in terms of vesicle size and drug release, obtained by encapsulation of FBX into the emulsomes (FBX-EMLs), was evaluated for its cytotoxic potential in human colorectal carcinoma (HCT 116) cells. The optimized FBX-EMLs formula had an improved half maximal inhibitory concentration (IC50), about 4-fold lower, compared to the free drug. The cell cycle analysis showed a significant inhibition of the HCT 116 cells proliferation following FBX-EMLs treatment compared to all the other conditions, with a higher number of cells accumulating on G2/M and pre-G1 phases, paralleled by a significant reduction of cells in G0/G1 and S phases. The optimized formula was also able to significantly increase the percentage of cell population in both early and late stages of apoptosis, characterized by a higher intracellular caspase-3 concentration, as well as percentage of necrotic cells. Lastly, the FBX ability to decrease the mitochondrial membrane potential was enhanced when the drug was delivered into the EMLs. In conclusion, the new formulation of FBX into EMLs improved all the parameters related to the anti-proliferative activity and the toxic potential of the drug towards colorectal cancer cells.
Multivariate Analytical Approaches to Identify Key Molecular Properties of Vehicles, Permeants and Membranes That Affect Permeation through Membranes
Authors: Najib Omaima N. , Kirton Stewart B. , Martin Gary P. , Botha Michelle J. , Sallam Al-Sayed , Murnane Darragh
Published: October 2020

There has been considerable recent interest in employing computer models to investigate the relationship between the structure of a molecule and its dermal penetration. Molecular permeation across the epidermis has previously been demonstrated to be determined by a number of physicochemical properties, for example, the lipophilicity, molecular weight and hydrogen bonding ability of the permeant. However little attention has been paid to modeling the combined effects of permeant properties in tandem with the properties of vehicles used to deliver those permeants or to whether data obtained using synthetic membranes can be correlated with those obtained using human epidermis. This work uses Principal Components Analysis (PCA) to demonstrate that, for studies of the diffusion of three model permeants (caffeine, methyl paraben and butyl paraben) through synthetic membranes, it is the properties of the oily vehicle in which they are applied that dominated the rates of permeation and flux. Simple robust and predictive descriptor-based quantitative structure–permeability relationship (QSPR) models have been developed to support these findings by utilizing physicochemical descriptors of the oily vehicles to quantify the differences in flux and permeation of the model compounds. Interestingly, PCA showed that, for the flux of co-applied model permeants through human epidermis, the permeation of the model permeants was better described by a balance between the physicochemical properties of the vehicle and the permeant rather than being dominated solely by the vehicle properties as in the case of synthetic model membranes. The important influence of permeant solubility in the vehicle along with the solvent uptake on overall permeant diffusion into the membrane was substantiated. These results confirm that care must be taken in interpreting permeation data when synthetic membranes are employed as surrogates for human epidermis; they also demonstrate the importance of considering not only the permeant properties but also those of both vehicle and membrane when arriving at any conclusions relating to permeation data.
Cellular Analysis and Chemotherapeutic Potential of a Bi-Functionalized Halloysite Nanotube
Authors: Luo Yangyang , Humayun Ahmed , Murray Teresa A. , Kemp Benjamin S. , McFarland Antwine , Liu Xuan , Mills David K.
Published: October 2020

The surface of halloysite nanotubes (HNTs) was bifunctionalized with two ligands—folic acid and a fluorochrome. In tandem, this combination should selectively target cancer cells and provide a means for imaging the nanoparticle. Modified bi-functionalized HNTs (bi-HNTs) were then doped with the anti-cancer drug methotrexate. bi-HNTs were characterized and subjected to in vitro tests to assess cellular growth and changes in cellular behavior in three cell lines—colon cancer, osteosarcoma, and a pre-osteoblast cell line (MC3T3-E1). Cell viability, proliferation, and cell uptake efficiency were assessed. The bi-HNTs showed cytocompatibility at a wide range of concentrations. Compared with regular-sized HNTs, reduced HNTs (~6 microns) were taken up by cells in more significant amounts, but increased cytotoxicity lead to apoptosis. Multi-photon images confirmed the intracellular location of bi-HNTs, and the method of cell entry was mainly through caveolae-mediated endocytosis. The bi-HNTs showed a high drug loading efficiency with methotrexate and a prolonged period of release. Most importantly, bi-HNTs were designed as a drug carrier to target cancer cells specifically, and imaging data shows that non-cancerous cells were unaffected after exposure to MTX-doped bi-HNTs. All data provide support for our nanoparticle design as a mechanism to selectively target cancer cells and significantly reduce the side-effects caused by off-targeting of anti-cancer drugs.
Microbubbles and Nanobubbles with Ultrasound for Systemic Gene Delivery
Authors: Endo-Takahashi Yoko , Negishi Yoichi
Published: October 2020

The regulation of gene expression is a promising therapeutic approach for many intractable diseases. However, its use in clinical applications requires the efficient delivery of nucleic acids to target tissues, which is a major challenge. Recently, various delivery systems employing physical energy, such as ultrasound, magnetic force, electric force, and light, have been developed. Ultrasound-mediated delivery has particularly attracted interest due to its safety and low costs. Its delivery effects are also enhanced when combined with microbubbles or nanobubbles that entrap an ultrasound contrast gas. Furthermore, ultrasound-mediated nucleic acid delivery could be performed only in ultrasound exposed areas. In this review, we summarize the ultrasound-mediated nucleic acid systemic delivery system, using microbubbles or nanobubbles, and discuss its possibilities as a therapeutic tool.
Novel Intrinsic Mechanisms of Active Drug Extrusion at the Blood-Brain Barrier: Potential Targets for Enhancing Drug Delivery to the Brain?
Authors: Löscher Wolfgang , Gericke Birthe
Published: October 2020

The blood-brain barrier (BBB) limits the pharmacotherapy of several brain disorders. In addition to the structural and metabolic characteristics of the BBB, the ATP-driven, drug efflux transporter P-glycoprotein (Pgp) is a selective gatekeeper of the BBB; thus, it is a primary hindrance to drug delivery into the brain. Here, we review the complex regulation of Pgp expression and functional activity at the BBB with an emphasis on recent studies from our laboratory. In addition to traditional processes such as transcriptional regulation and posttranscriptional or posttranslational modification of Pgp expression and functionality, novel mechanisms such as intra- and intercellular Pgp trafficking and intracellular Pgp-mediated lysosomal sequestration in BBB endothelial cells with subsequent disposal by blood neutrophils are discussed. These intrinsic mechanisms of active drug extrusion at the BBB are potential therapeutic targets that could be used to modulate P-glycoprotein activity in the treatment of brain diseases and enhance drug delivery to the brain.
Rapid Target Binding and Cargo Release of Activatable Liposomes Bearing HER2 and FAP Single-Chain Antibody Fragments Reveal Potentials for Image-Guided Delivery to Tumors
Authors: Tansi Felista L. , Rüger Ronny , Böhm Claudia , Steiniger Frank , Raasch Martin , Mosig Alexander S. , Kontermann Roland E. , Teichgräber Ulf K. , Fahr Alfred , Hilger Ingrid
Published: October 2020

Liposomes represent suitable tools for the diagnosis and treatment of a variety of diseases, including cancers. To study the role of the human epidermal growth factor receptor 2 (HER2) as target in cancer imaging and image-guided deliveries, liposomes were encapsulated with an intrinsically quenched concentration of a near-infrared fluorescent dye in their aqueous interior. This resulted in quenched liposomes (termed LipQ), that were fluorescent exclusively upon degradation, dye release, and activation. The liposomes carried an always-on green fluorescent phospholipid in the lipid layer to enable tracking of intact liposomes. Additionally, they were functionalized with single-chain antibody fragments directed to fibroblast activation protein (FAP), a marker of stromal fibroblasts of most epithelial cancers, and to HER2, whose overexpression in 20–30% of all breast cancers and many other cancer types is associated with a poor treatment outcome and relapse. We show that both monospecific (HER2-IL) and bispecific (Bi-FAP/HER2-IL) formulations are quenched and undergo HER2-dependent rapid uptake and cargo release in cultured target cells and tumor models in mice. Thereby, tumor fluorescence was retained in whole-body NIRF imaging for 32–48 h post-injection. Opposed to cell culture studies, Bi-FAP/HER2-IL-based live confocal microscopy of a high HER2-expressing tumor revealed nuclear delivery of the encapsulated dye. Thus, the liposomes have potentials for image-guided nuclear delivery of therapeutics, and also for intraoperative delineation of tumors, metastasis, and tumor margins.
Deep Tumor Penetration of Doxorubicin-Loaded Glycol Chitosan Nanoparticles Using High-Intensity Focused Ultrasound
Authors: Choi Yongwhan , Han Hyounkoo , Jeon Sangmin , Yoon Hong Yeol , Kim Hyuncheol , Kwon Ick Chan , Kim Kwangmeyung
Published: October 2020

The dense extracellular matrix (ECM) in heterogeneous tumor tissues can prevent the deep tumor penetration of drug-loaded nanoparticles, resulting in a limited therapeutic efficacy in cancer treatment. Herein, we suggest that the deep tumor penetration of doxorubicin (DOX)-loaded glycol chitosan nanoparticles (CNPs) can be improved using high-intensity focused ultrasound (HIFU) technology. Firstly, we prepared amphiphilic glycol chitosan-5β-cholanic acid conjugates that can self-assemble to form stable nanoparticles with an average of 283.7 ± 5.3 nm. Next, the anticancer drug DOX was simply loaded into the CNPs via a dialysis method. DOX-loaded CNPs (DOX-CNPs) had stable nanoparticle structures with an average size of 265.9 ± 35.5 nm in aqueous condition. In cultured cells, HIFU-treated DOX-CNPs showed rapid drug release and enhanced cellular uptake in A549 cells, resulting in increased cytotoxicity, compared to untreated DOX-CNPs. In ECM-rich A549 tumor-bearing mice, the tumor-targeting efficacy of intravenously injected DOX-CNPs with HIFU treatment was 1.84 times higher than that of untreated DOX-CNPs. Furthermore, the deep tumor penetration of HIFU-treated DOX-CNPs was clearly observed at targeted tumor tissues, due to the destruction of the ECM structure via HIFU treatment. Finally, HIFU-treated DOX-CNPs greatly increased the therapeutic efficacy at ECM-rich A549 tumor-bearing mice, compared to free DOX and untreated DOX-CNPs. This deep penetration of drug-loaded nanoparticles via HIFU treatment is a promising strategy to treat heterogeneous tumors with dense ECM structures.
Viral load Reduction in SHIV-Positive Nonhuman Primates via Long-Acting Subcutaneous Tenofovir Alafenamide Fumarate Release from a Nanofluidic Implant
Authors: Pons-Faudoa Fernanda P. , Trani Nicola Di , Sizovs Antons , Shelton Kathryn A. , Momin Zoha , Bushman Lane R. , Xu Jiaqiong , Lewis Dorothy E. , Demaria Sandra , Hawkins Trevor , Rooney James F. , Marzinke Mark A. , Kimata Jason T. , Anderson Peter L. , Nehete Pramod N. , Arduino Roberto C. , Sastry K. Jagannadha , Grattoni Alessandro
Published: October 2020

HIV-1 is a chronic disease managed by strictly adhering to daily antiretroviral therapy (ART). However, not all people living with HIV-1 have access to ART, and those with access may not adhere to treatment regimens increasing viral load and disease progression. Here, a subcutaneous nanofluidic implant was used as a long-acting (LA) drug delivery platform to address these issues. The device was loaded with tenofovir alafenamide (TAF) and implanted in treatment-naïve simian HIV (SHIV)-positive nonhuman primates (NHP) for a month. We monitored intracellular tenofovir-diphosphate (TFV-DP) concentration in the target cells, peripheral blood mononuclear cells (PBMC). The concentrations of TFV-DP were maintained at a median of 391.0 fmol/106 cells (IQR, 243.0 to 509.0 fmol/106 cells) for the duration of the study. Further, we achieved drug penetration into lymphatic tissues, known for persistent HIV-1 replication. Moreover, we observed a first-phase viral load decay of −1.14 ± 0.81 log10 copies/mL (95% CI, −0.30 to −2.23 log10 copies/mL), similar to −1.08 log10 copies/mL decay observed in humans. Thus, LA TAF delivered from our nanofluidic implant had similar effects as oral TAF dosing with a lower dose, with potential as a platform for LA ART.
Sustained Release of Linezolid from Prepared Hydrogels with Polyvinyl Alcohol and Aliphatic Dicarboxylic Acids of Variable Chain Lengths
Authors: Carreño Gustavo , Marican Adolfo , Vijayakumar Sekar , Valdés Oscar , Cabrera-Barjas Gustavo , Castaño Johanna , Durán-Lara Esteban F.
Published: October 2020

A series of hydrogels with a specific release profile of linezolid was successfully synthesized. The hydrogels were synthesized by cross-linking polyvinyl alcohol (PVA) and aliphatic dicarboxylic acids, which include succinic acid (SA), glutaric acid (GA), and adipic acid (AA). The three crosslinked hydrogels were prepared by esterification and characterized by equilibrium swelling ratio, infrared spectroscopy, thermogravimetric analysis, mechanical properties, and scanning electron microscopy. The release kinetics studies of the linezolid from prepared hydrogels were investigated by cumulative drug release and quantified by chromatographic techniques. Mathematical models were carried out to understand the behavior of the linezolid release. These data revealed that the sustained release of linezolid depends on the aliphatic dicarboxylic acid chain length, their polarity, as well as the hydrogel crosslinking degree and mechanical properties. The in vitro antibacterial assay of hydrogel formulations was assessed in an Enterococcus faecium bacterial strain, showing a significant activity over time. The antibacterial results were consistent with cumulative release assays. Thus, these results demonstrated that the aliphatic dicarboxylic acids used as crosslinkers in the PVA hydrogels were a determining factor in the antibiotic release profile.
Root System Architecture, Copper Uptake and Tissue Distribution in Soybean (Glycine max (L.) Merr.) Grown in Copper Oxide Nanoparticle (CuONP)-Amended Soil and Implications for Human Nutrition
Authors: Yusefi-Tanha Elham , Fallah Sina , Rostamnejadi Ali , Pokhrel Lok Raj
Published: October 2020

Understanding the potential uptake and biodistribution of engineered nanoparticles (ENPs) in soil-grown plants is imperative for realistic toxicity and risk assessment considering the oral intake of edibles by humans. Herein, growing N-fixing symbiont (Bradyrhizobium japonicum) inoculated soybean (Glycine max (L.) Merr.) for a full lifecycle of 120 days, we assessed the potential influence of particle size (25, 50, and 250 nm) and concentration (0, 50, 100, 200, and 500 mg/kg soil) of Copper oxide nanoparticles (CuONPs) on: (1) root system architecture, (2) soil physicochemical attributes at the soil–root interface, and (3) Cu transport and accumulation in root, stem, leaf, and seed in soybean, and compared them with the soluble Cu2+ ions and water-only controls. Finally, we performed a comparative assessment of total seed Cu levels in soybean with other valuable food sources for Cu intake and discussed potential human health implications. Results showed particle size- and concentration-dependent influence of CuONPs on Cu uptake and distribution in root, stem, leaf, and seed. Alterations in root architecture (root biomass, length, volume, and area) were dependent on the Cu compound types, Cu concentrations, and their interactions. Concentration–response relationships for all three sizes of CuONPs and Cu2+ ions were found to be linear. Furthermore, CuONPs and Cu2+ ions had inhibitory effects on root growth and development. Overall, soybean responses to the smallest size of CuONPs–25 nm—were greater for all parameters tested compared to the two larger-sized CuONPs (50 nm, 250 nm) or Cu2+ ions. Results suggest that minor changes in soil-root physicochemical attributes may not be a major driver for Cu uptake in soybean. Cu bioaccumulation followed the order: root > leaf > stem > seed. Despite reduction in root architecture and seed yield, the smallest size CuONPs–25 nm led to increased total seed Cu uptake compared to the larger-sized CuONPs or Cu2+ ions. Our findings also suggest that soil amendment with CuONPs, and more so with the smallest size of CuONPs–25 nm—could significantly improve seed nutritional Cu value in soybean as reflected by the % Daily Values (DV) and are rated “Good” to “Very Good” according to the “World’s Healthiest Foods” rating. However, until the potential toxicity and risk from CuONP-fortified soybean seed ingestion is characterized in humans, we caution recommending such seeds for daily human consumption when addressing food Cu-deficiency and associated diseases, globally.
Early Detection of Airborne Inoculum of Nothopassalora personata in Spore Trap Samples from Peanut Fields Using Quantitative PCR
Authors: Munir Misbakhul , Wang Hehe , Dufault Nicholas S. , Anco Daniel J.
Published: October 2020

A quantitative PCR (qPCR)-assay was developed to detect airborne inoculum of Nothopassalora personata, causal agent of late leaf spot (LLS) on peanut, collected with a modified impaction spore trap. The qPCR assay was able to consistently detect as few as 10 spores with purified DNA and 25 spores based on crude DNA extraction from rods. In 2019, two spore traps were placed in two peanut fields with a history of LLS. Sampling units were replaced every 2 to 4 days and tested with the developed qPCR assay, while plots were monitored for symptom development. The system detected inoculum 35 to 56 days before visual symptoms developed in the field, with detection related to environmental parameters affecting pathogen life-cycle and disease development. This study develops the framework of the qPCR spore trap system and represents the initial steps towards validation of the performance of the system for use as a decision support tool to complement integrated management of LLS.
Phytotoxic Activity of the Natural Compound Norharmane on Crops, Weeds and Model Plants
Authors: López-González David , Ledo David , Cabeiras-Freijanes Luz , Verdeguer Mercedes , Reigosa Manuel , Sánchez-Moreiras Adela
Published: October 2020

Norharmane is a secondary metabolite that appears in different species of land plants. In this paper, we investigated for the first time the specificity of norharmane through germination and growth tests on some crops as Zea mays L. (maize), Triticum aestivum L. (wheat), Oryza sativa L. (rice) and Lactuca sativa L. (lettuce) and weeds as Amaranthus retroflexus L. (amaranth), Echinochloa crus-galli L. (barnyard grass), Plantago lanceolata L. (ribwort), Portulaca oleracea L. (common purslane) and Avena fatua L. (wild oat), and its phytotoxic capacity on the metabolism of adult Arabidopsis thaliana L. (thale cress) by measuring chlorophyll a fluorescence, pigment content, total proteins, osmotic potential and morphological analysis. Norharmane had an inhibitory effect on the germination of A. fatua and P. lanceolata, and the growth of P. oleracea, E. crus-galli and A. retroflexus. On adult A. thaliana plants, the compound was more effective to watering, leading to water stress that compromised the growth of the plants and ultimately affected the photosynthetic apparatus. Therefore, this research shows that norharmane not only affects seedlings’ metabolism, but also damages the metabolism of adult plants and can be a potential model for a future bioherbicide given its specificity.