bluuebunny/crossref_metadata_2025 huggingface dataset:
SPME-GC/MS Analysis of Methanol in Biospecimen by Derivatization with Pyran Compound
Authors: Lee Joon-Bae , Jeong Yong Ae , Ahn Dae Jun , Bang Iel Soo
Published: December 2019

Methanol is metabolized in the body to highly toxic formaldehyde and formate when consumed accidentally. Methanol has been typically analyzed with gas chromatography-flame ionization detector (GC-FID). However, its retention time may overlap with other volatile compounds and lead to confusion. Alternative analysis of methanol using gas chromatography/mass spectrometry (GC/MS) also has limitations due to its similar molecular weight with oxygen and low boiling point. In this study, methanol and internal standard of deuterium-substituted ethanol were derivatized with 3,4-dihydro-2H-pyran under acid catalysis using concentrated hydrochloric acid. The reaction products including 2-methoxytetrahydropyran were extracted with solid-phase microextraction followed by GC/MS analysis. This method was successfully applied to measure the lethal concentration of methanol in the blood of a victim with a standard addition method to overcome the complex matrix effect of the biospecimen. Identification of the metabolite formate by ion chromatography confirmed the death cause to be methanol poisoning. This new method was a much more convenient and reliable process to measure methanol in complex matrix samples by reducing sample pretreatment effort and cost.
Indole-Containing Phytoalexin-Based Bioisosteres as Antifungals: In Vitro and In Silico Evaluation against Fusarium oxysporum
Authors: Angarita-Rodríguez Andrea , Quiroga Diego , Coy-Barrera Ericsson
Published: December 2019

There is a continuous search for more reliable and effective alternatives to control phytopathogens through different strategies. In this context, indole-containing phytoalexins are stimuli-induced compounds implicated in plant defense against plant pathogens. However, phytoalexins’ efficacy have been limited by fungal detoxifying mechanisms, thus, the research on bioisosteres-based analogs can be a friendly alternative regarding the control of Fusarium phytopathogens, but there are currently few studies on it. Thus, as part of our research on antifungal agents, a set of 21 synthetic indole-containing phytoalexin analogs were evaluated as inhibitors against the phyopathogen Fusarium oxysporum. Results indicated that analogs of the N,N-dialkylthiourea, N,S-dialkyldithiocarbamate and substituted-1,3-thiazolidin-5-one groups exhibited the best docking scores and interaction profiles within the active site of Fusarium spp. enzymes. Vina scores exhibited correlation with experimental mycelial growth inhibition using supervised statistics, and this antifungal dataset correlated with molecular interaction fields after CoMFA. Compound 24 (tert-butyl (((3-oxo-1,3-diphenylpropyl)thio)carbonothioyl)-l-tryptophanate), a very active analog against F. oxysporum, exhibited the best interaction with lanosterol 14α-demethylase according to molecular docking, molecular dynamics and molecular mechanic/poisson-boltzmann surface area (MM/PBSA) binding energy performance. After data analyses, information on mycelial growth inhibitors, structural requirements and putative enzyme targets may be used in further antifungal development based on phytoalexin analogs for controlling phytopathogens.
Design and Biological Evaluation of Colchicine-CD44-Targeted Peptide Conjugate in an In Vitro Model of Crystal Induced Inflammation
Authors: Zoghebi Khalid A. , Bousoik Emira , Parang Keykavous , Elsaid Khaled A.
Published: December 2019

Gout is an inflammatory arthritis due to the joint deposition of monosodium urate (MSU) crystals. Phagocytosis of MSU crystals by tissue macrophages results in the generation of reactive oxygen species (ROS) and production of inflammatory cytokines and chemokines. Colchicine use in gout is limited by severe toxicity. CD44 is a transmembrane glycoprotein that is highly expressed in tissue macrophages and may be involved in gout pathogenesis. The P6 peptide is a 20-amino acid residue peptide that binds to CD44. We hypothesized that the conjugation of colchicine to the P6 peptide would reduce its off-target cytotoxicity while preserving its anti-inflammatory effect. A modified version of P6 peptide and colchicine-P6 peptide conjugate were synthesized using Fmoc/tBu solid-phase and solution-phase chemistry, respectively. A glutaryl amide was used as a linker. The P6 peptide was evaluated for its binding to CD44, association, and internalization by macrophages. Cytotoxic effects of P6 peptide, colchicine, and colchicine-P6 peptide on macrophages were compared and the inhibition of ROS generation and interleukin-8 (IL-8) secretion in MSU-stimulated macrophages treated with P6 peptide, colchicine, or colchicine-P6 peptide was studied. We confirmed that the P6 peptide binds to CD44 and its association and internalization by macrophages were CD44-dependent. Colchicine (1, 10, and 25 μM) demonstrated a significant cytotoxic effect on macrophages while the P6 peptide and colchicine-P6 peptide conjugate (1, 10 and 25 μM) did not alter the viability of the macrophages. The P6 peptide (10 and 25 μM) reduced ROS generation and IL-8 secretion mediated by a reduction in MSU phagocytosis by macrophages. The colchicine-P6 peptide significantly reduced ROS generation and IL-8 secretion compared to the P6 peptide alone at 1 and 10 μM concentrations. Conjugation of colchicine to the P6 peptide reduced the cytotoxic effect of colchicine while preserving its anti-inflammatory activity.
Preparation of Activated Carbon Supported Bead String Structure Nano Zero Valent Iron in a Polyethylene Glycol-Aqueous Solution and Its Efficient Treatment of Cr(VI) Wastewater
Authors: Jiao Chunlei , Tan Xiao , Lin Aijun , Yang Wenjie
Published: December 2019

Nanometer zero-valent iron (nZVI) has been widely used in the treatment of heavy metals such as hexavalent chromium (Cr(VI)). A novel composite of bead string-structured nZVI on modified activated carbon (nZVI–MAC) is prepared here, using polyethylene glycol as the stable dispersant rather than traditional ethanol during the loading process. The microstructure characterization shows that nZVI particles are loaded on MAC with a bead string structure in large quantity and stably due to the addition of hydroxyl functional groups on the surface by polyethylene glycol. Experiments on the treatment of Cr(VI) in wastewater show that the reaction process requires only 20 min to achieve equilibrium. The removal rate of Cr(VI) with a low concentration (80–100 mg/L) is over 99% and the maximum saturation removal capacity is up to 66 mg/g. The system converts Cr(VI) to trivalent chromium (Cr(III)) through an oxidation-reduction effect and forms an insoluble material with iron ions by coprecipitation, which is then adsorbed on the surface of the nZVI–MAC. The process conforms to the quasi-second order adsorption kinetics equation (mainly chemical adsorption process).
Development of a Direct Competitive ELISA Kit for Detecting Deoxynivalenol Contamination in Wheat
Authors: Han Li , Li Yue-Tao , Jiang Jin-Qing , Li Ren-Feng , Fan Guo-Ying , Lv Jun-Mei , Zhou Ye , Zhang Wen-Ju , Wang Zi-Liang
Published: December 2019

This study was conducted to develop a self-assembled direct competitive enzyme-linked immunosorbent assay (dcELISA) kit for the detection of deoxynivalenol (DON) in food and feed grains. Based on the preparation of anti-DON monoclonal antibodies, we established a standard curve with dcELISA and optimized the detection conditions. The performance of the kit was evaluated by comparison with high-performance liquid chromatography (HPLC). The minimum detection limit of DON with the kit was 0.62 ng/mL, the linear range was from 1.0 to 113.24 ng/mL and the half-maximal inhibition concentration (IC50) was 6.61 ng/mL in the working buffer; there was a limit of detection (LOD) of 62 ng/g, and the detection range was from 100 to 11324 ng/g in authentic agricultural samples. We examined four samples of wheat bran, wheat flour, corn flour and corn for DON recovery. The average recovery was in the range of 77.1% to 107.0%, and the relative standard deviation (RSD) ranged from 4.2% to 11.9%. In addition, the kit has the advantages of high specificity, good stability, a long effective life and negligible sample matrix interference. Finally, wheat samples from farms in the six provinces of Henan, Anhui, Hebei, Shandong, Jiangsu and Gansu in China were analyzed by the kit. A total of 30 samples were randomly checked (five samples in each province), and the results were in good agreement with the standardized HPLC method. These tests showed that the dcELISA kit had good performance and met relevant technical requirements, and it had the characteristics of accuracy, reliability, convenience and high-throughput screening for DON detection. Therefore, the developed kit is suitable for rapid screening of DON in marketed products.
Ascorbic Acid-Assisted Polyol Synthesis of Iron and Fe/GO, Fe/h-BN Composites for Pb2+ Removal from Wastewaters
Authors: Leybo Denis , Tagirov Marat , Permyakova Elizaveta , Konopatsky Anton , Firestein Konstantin , Tuyakova Feruza , Arkhipov Dmitry , Kuznetsov Denis
Published: December 2019

Iron powders and Fe/graphene oxide and Fe/boron nitride composites were synthesized by means of a polyol synthesis method. The effect of NaOH/Fe and ascorbic acid/Fe ratios on the characteristics of synthesized products were evaluated. The samples were characterized by X-ray diffraction, scanning and transmission electron microscopy, low-temperature nitrogen adsorption and Raman-spectroscopy. Ascorbic acid-assisted polyol synthesis resulted in the 10-fold decrease of the iron particles’ size and almost 2-fold increase of lead removal efficiency. The deposition of iron on the surface of graphene oxide lead to the formation of small 20–30 nm sized particles as well as bigger 200–300 nm sized particles, while the reduction in presence of boron nitride resulted in the 100–200 nm sized particles. The difference is attributed to the surface state of graphene oxide and boron nitride. Adsorption properties of the obtained materials were studied in the process of Pb2+ ion removal from wastewater.
Correction: Schraders, K. et al. Quantitative Ultrasound and Dual X-ray Absorptiometry as Indicators of Bone Mineral Density in Young Women and Nutritional Factors Affecting It. Nutrients, 2019, 11, 2336
Authors: Schraders Katie , Zatta Giancarla , Kruger Marlena , Coad Jane , Weber Janet , Brough Louise , Thomson Jasmine
Published: December 2019

The authors would like to make the following correction to our recent publication [...]
Effects of Nutritional Education Interventions on Metabolic Risk in Children and Adolescents: A Systematic Review of Controlled Trials
Authors: Leis Rosaura , de Lamas Carmela , de Castro María-José , Picáns Rosaura , Gil-Campos Mercedes , Couce María L.
Published: December 2019

Childhood obesity is a global public health issue and is linked to metabolic syndrome, which increases the risk of comorbidities such as type 2 diabetes, cardiovascular diseases and cancer. Social, economic and cultural factors influence changes in nutrition and lifestyle characterized by poorer diets and reduced physical activity. This systematic review summarizes the evidence for nutritional education interventions to improve metabolic risks in children and adolescents. Systematic searches of the databases Medline (via PubMed) and Scopus were conducted following PRISMA guidelines. The risk of bias for each study was assessed following the methodology of the Cochrane Collaboration. Ten case-controlled and randomized controlled studies testing nutritional educational interventions targeting children and adolescents from the general population were eligible for inclusion. The sample size was 3915 and the age range was 7–20 years. The duration of intervention ranged from 12 weeks to 20 years. All the studies that provided data on abdominal obesity reported differences in favour of the intervention. However, data on the effects on the remaining components of metabolic syndrome remain inconclusive. These results support the role of nutritional education interventions as a strategy to reduce central adiposity and its possible unhealthy consequences in children and adolescents.
Milk Consumption Decreases Risk for Breast Cancer in Korean Women under 50 Years of Age: Results from the Health Examinees Study
Authors: Shin Woo-Kyoung , Lee Hwi-Won , Shin Aesun , Lee Jong-koo , Kang Daehee
Published: December 2019

Epidemiologic studies regarding breast cancer risk related to milk consumption remain controversial. The aim of this study was to evaluate the association between milk consumption and the risk for breast cancer. A total of 93,306 participants, aged 40–69 years, were included in the prospective cohort study in the Health Examinees-Gem (HEXA-G) study between 2004 and 2013. Dietary intake was assessed using a validated food frequency questionnaire. Information on cancer diagnosis in the eligible cohort was retrieved from the Korea Central Cancer Registry through 31 December 2014. The Cox proportional hazards model was used to estimate multivariate hazard ratios (HRs) and 95% confidence intervals (CIs). A total of 359 breast cancer cases were observed over a median follow-up period of 6.3 years. Milk consumption was not associated with decreased risk for breast cancer in the total population (p for trend = 0.0687). In women under 50 years of age, however, milk consumption was inversely associated with breast cancer risk. In the comparison between highest (≥1 serving/day) and lowest (<1 serving/week) intake categories of milk, the multivariate HR (95% CI) was 0.58 (0.35–0.97, p for trend = 0.0195)) among women under 50 years of age. In conclusion, our findings show that milk consumption in Korean women aged 50 or younger is associated with a decreased risk for breast cancer, when compared to those who never or rarely consumed milk. Further studies need to be conducted to assess this relationship and confirm these results.
The Mediterranean Diet Slows Down the Progression of Aging and Helps to Prevent the Onset of Frailty: A Narrative Review
Authors: Capurso Cristiano , Bellanti Francesco , Lo Buglio Aurelio , Vendemiale Gianluigi
Published: December 2019

The aging population is rapidly increasing all over the world. This results in significant implications for the planning and provision of health and social care. Aging is physiologically characterized by a decrease in lean mass, bone mineral density and, to a lesser extent, fat mass. The onset of sarcopenia leads to weakness and a further decrease in physical activity. An insufficient protein intake, which we often observe in patients of advanced age, certainly accelerates the progression of sarcopenia. In addition, many other factors (e.g., insulin resistance, impaired protein digestion and absorption of amino acids) reduce the stimulation of muscle protein synthesis in the elderly, even if the protein intake is adequate. Inadequate intake of foods can also cause micronutrient deficiencies that contribute to the development of frailty. We know that a healthy eating style in middle age predisposes to so-called “healthy and successful” aging, which is the condition of the absence of serious chronic diseases or of an important decline in cognitive or physical functions, or mental health. The Mediterranean diet is recognized to be a “healthy food” dietary pattern; high adherence to this dietary pattern is associated with a lower incidence of chronic diseases and lower physical impairment in old age. The aim of our review was to analyze observational studies (cohort and case–control studies) that investigated the effects of following a healthy diet, and especially the effect of adherence to a Mediterranean diet (MD), on the progression of aging and on onset of frailty.
Leptin, Ghrelin, and Leptin/Ghrelin Ratio in Critically Ill Patients
Authors: Arabi Yaseen M. , Jawdat Dunia , Al-Dorzi Hasan M. , Tamim Hani , Tamimi Waleed , Bouchama Abderrezak , Sadat Musharaf , Afesh Lara , Abdullah Mashan L. , Mashaqbeh Walid , Sakhija Maram , Al-Dawood Abdulaziz
Published: December 2019

The objective of this study was to evaluate leptin, ghrelin, and leptin/ghrelin ratio in critically ill patients and association of leptin/ghrelin ratio with outcomes. This is a sub-study of the PermiT trial (ISRCTN68144998). A subset of 72 patients who were expected to stay >14 days in the Intensive care unit were enrolled. Blood samples were collected on days 1, 3, 5, 7, and 14. Samples were analyzed for leptin and active ghrelin in addition to other hormones. Baseline leptin/ghrelin ratio was calculated, and patients were stratified into low and high leptin/ghrelin ratio based on the median value of 236. There was a considerable variation in baseline leptin level: Median 5.22 ng/mL (Q1, Q3: 1.26, 17.60). Ghrelin level was generally low: 10.61 pg/mL (Q1, Q3: 8.62, 25.36). Patients with high leptin/ghrelin ratio compared to patients with low leptin/ghrelin ratio were older, had higher body mass index and more likely to be diabetic. There were no differences in leptin/ghrelin ratio between patients who received permissive underfeeding and standard feeding. Multivariable logistic regression analysis showed that age and body mass index were significant independent predictors of high leptin–ghrelin ratio. Leptin–ghrelin ratio was not associated with 90-day mortality or other outcomes. Age and body mass index are predictors of high leptin/ghrelin ratio. Leptin/ghrelin ratio is not affected by permissive underfeeding and is not associated with mortality.
Observed Dietary Intake in Adults with Intellectual Disability Living in Group Homes
Authors: Hamzaid Nur Hana , O’Connor Helen T. , Flood Victoria M.
Published: December 2019

Background: There is limited information on the dietary intakes of people with intellectual disability (ID) living in group homes. Objective: To describe and evaluate dietary intake in people with ID. Method: Dietary intake was assessed in a convenience sample of people with ID living in group homes. Dietary assessment used three-day weighed food records and digital food photography. Intakes were compared to the Nutrient Reference Values (NRVs) and dietary recommendations. Results: A sample of 33 adults, (men (M): n = 14; women (W): n = 19), mean age 51 ± 14 years, was recruited from seven group homes. Mean daily energy intake was low (M: 7.4 MJ; W: 7.0 MJ; p = 0.46), similar to levels recommended for bed rest. Many participants had intakes below the estimated average requirements (EARs) for the nutrients, magnesium (M: 86%; W: 63%), calcium (M: 43%; W: 78%), iodine (M: 43%; W: 47%) and zinc (M: 43%). Less than half of the recommended daily servings were consumed for vegetables (men and women) and dairy foods (women). Conclusion: Nutrient intake and diet quality of the participants in the group homes studied was poor. Education and policy to support healthier diets is required to improve dietary intake of people with intellectual disability, living in group homes.
Switching to a Healthy Diet Prevents the Detrimental Effects of Western Diet in a Colitis-Associated Colorectal Cancer Model
Authors: Gröschel Charlotte , Prinz-Wohlgenannt Maximilian , Mesteri Ildiko , Karuthedom George Sobha , Trawnicek Lena , Heiden Denise , Aggarwal Abhishek , Tennakoon Samawansha , Baumgartner Maximilian , Gasche Christoph , Lang Michaela , Marculescu Rodrig , Manhardt Teresa , Schepelmann Martin , Kallay Enikö
Published: December 2019

Inflammatory bowel disease increases the odds of developing colitis-associated cancer. We hypothesized that Western-style diet (WD) aggravates azoxymethane (AOM)/dextran sulfate sodium salt (DSS)-induced colitis-associated tumorigenesis and that switching to the standard AIN93G diet will ameliorate disease symptoms even after cancer initiation. Female BALB/c mice received either WD (WD group) or standard AIN93G diet (AIN group) for the whole experimental period. After five weeks, the mice received 12.5 mg/kg AOM intraperitoneally, followed by three DSS cycles. In one group of mice, the WD was switched to AIN93G the day before starting the first DSS cycle (WD/AIN group). Feeding the WD during the whole experimental period aggravated colitis symptoms, shortened the colon (p < 0.05), changed microbiota composition and increased tumor promotion. On molecular level, the WD reduced proliferation (p < 0.05) and increased expression of the vitamin D catabolizing enzyme Cyp24a1 (p < 0.001). The switch to the AIN93G diet ameliorated this effect, reflected by longer colons, fewer (p < 0.05) and smaller (p < 0.01) aberrant colonic crypt foci, comparable with the AIN group. Our results show that switching to a healthy diet, even after cancer initiation is able to revert the deleterious effect of the WD and could be an effective preventive strategy to reduce colitis symptoms and prevent tumorigenesis.
Faecal Indicator Bacteria and Pseudomonas aeruginosa in Marine Coastal Waters: Is there a Relationship?
Authors: Januário Adriana P. , Afonso Clélia N. , Mendes Susana , Rodrigues Maria J.
Published: December 2019

To estimate the quality of coastal waters, European Union Directive 2006/7/EC provides guidelines to assess levels of faecal bacteria, including Escherichia coli and intestinal enterococci. These microbiological criteria are based on studies that determine the risk of bathers having diseases caused by enteric bacteria, not necessarily measuring the potential danger associated with the presence of nonenteric pathogens. The association between the presence of faecal contaminant indicators and nonenteric pathogenic microorganisms has not been well defined yet. The purpose of this study is to establish a relationship between Pseudomonas aeruginosa and microbiological indicators of faecal contamination. Presence of microbiological contamination in the coastal waters near the sewage treatment plant (STP) of Peniche (Portugal) was confirmed (P. aeruginosa 135.8 Colony Forming Unit/100 mL, Escherichia coli 1100.1 Most Probable Number/100 mL, intestinal enterococci 2685.9 MPN/100 mL) with much lower levels in the areas located south of the STP, along the main water coastal current (beach 1: 0.7 CFU/100 mL, 16.5 MPN/100 mL, 100.5 MPN/100 mL; beach 2: 0.3 CFU/100 mL, 74.0 MPN/100 mL, 145.9 MPN/100 mL, respectively). Analysis of Pearson’s correlation revealed a strong positive correlation between E. coli and P. aeruginosa, suggesting E. coli as an indicator of P. aeruginosa presence.
The Malacosporean Myxozoan Parasite Tetracapsuloides bryosalmonae: A Threat to Wild Salmonids
Authors: Sudhagar Arun , Kumar Gokhlesh , El-Matbouli Mansour
Published: December 2019

Tetracapsuloides bryosalmonae is a myxozoan parasite responsible for proliferative kidney disease (PKD) in a wide range of salmonids. PKD, characterized by high mortality and morbidity, is well known for affecting aquaculture operations and wild salmonid populations across Europe and North America. The life cycle of T. bryosalmonae revolves around freshwater bryozoan and salmonid fish hosts. In recent years, T. bryosalmonae has been reported among wild salmonids from the European countries where it has not been reported previously. T. bryosalmonae is believed to be a possible reason for the diminishing wild salmonid populations in the natural water bodies of many European countries. Climate crisis driven rising water temperature can further accelerate the distribution of T. bryosalmonae. Expansion of the geographical distribution of T. bryosalmonae may further advocate the decline of wild salmonid populations, especially brown trout (Salmo trutta) in their habitats. Mathematical models are used to understand the pattern and distribution of T. bryosalmonae among the host in the natural water bodies. The present manuscript not only summarizes the incidences of T. bryosalmonae among the wild salmonid populations, but also discusses the contemporary understanding about the development of T. bryosalmonae in its hosts and the influences of various factors in the spread of the disease in the wild.
Rational Design of a Multipurpose Bioadhesive Vaginal Film for Co-Delivery of Dapivirine and Levonorgestrel
Authors: Li Jing , Regev Galit , Patel Sravan Kumar , Patton Dorothy , Sweeney Yvonne , Graebing Philip , Grab Sheila , Wang Lin , Sant Vinayak , Rohan Lisa C.
Published: December 2019

Human immunodeficiency virus (HIV) infection and unintended pregnancy, which can lead to life-threatening complications, are two major burdens for female reproductive health. To address these pressing health issues, multipurpose prevention technologies (MPTs) are proposed to deliver two or more drugs simultaneously. MPTs could offer several benefits for users such as improved convenience, increased effectiveness, reduced cost, and decreased environmental burden. Here, we report the development, and in vitro and in vivo assessment of a bioadhesive vaginal film as a coitally-independent MPT dosage form for delivering dapivirine (DPV) and levonorgestrel (LNG) to prevent HIV infection and unintended pregnancy, respectively. After confirming the feasibility of bioadhesive film use for weekly drug delivery in vivo through colpophotography and MRI evaluation, the pharmacokinetics (PK) of DPV/LNG single entity and combination bioadhesive films was investigated in pigtailed macaques (n = 5). Both drugs from single entity or combination films were able to provide sustained drug release in vivo. The combination film showed lower local tissue clearance for DPV and exhibited significantly increased plasma concentration for LNG as compared to the single entity film. This proof-of-concept study demonstrates the ability of this novel bioadhesive film platform to deliver LNG and DPV simultaneously as an MPT product for the prevention of HIV infection and unintended pregnancy.
Quality by Design Micro-Engineering Optimisation of NSAID-Loaded Electrospun Fibrous Patches
Authors: Nazari Kazem , Mehta Prina , Arshad Muhammad Sohail , Ahmed Shahabuddin , Andriotis Eleftherios G. , Singh Neenu , Qutachi Omar , Chang Ming-Wei , Fatouros Dimitrios G. , Ahmad Zeeshan
Published: December 2019

The purpose of this study was to apply the Quality by Design (QbD) approach to the electrospinning of fibres loaded with the nonsteroidal anti-inflammatory drugs (NSAIDs) indomethacin (INDO) and diclofenac sodium (DICLO). A Quality Target Product Profile (QTPP) was made, and risk assessments (preliminary hazard analysis) were conducted to identify the impact of material attributes and process parameters on the critical quality attributes (CQAs) of the fibres. A full factorial design of experiments (DoE) of 20 runs was built, which was used to carry out experiments. The following factors were assessed: Drugs, voltage, flow rate, and the distance between the processing needle and collector. Release studies exhibited INDO fibres had greater total release of active drug compared to DICLO fibres. Voltage and distance were found to be the most significant factors of the experiment. Multivariate statistical analytical software helped to build six feasible design spaces and two flexible, universal design spaces for both drugs, at distances of 5 cm and 12.5 cm, along with a flexible control strategy. The current findings and their analysis confirm that QbD is a viable and invaluable tool to enhance product and process understanding of electrospinning for the assurance of high-quality fibres.
Fab-Arm Exchange Combined with Selective Protein A Purification Results in a Platform for Rapid Preparation of Monovalent Bispecific Antibodies Directly from Culture Media
Authors: Steinhardt James , Wu Yanli , Fleming Ryan , Ruddle Ben T. , Patel Pooja , Wu Herren , Gao Changshou , Dimasi Nazzareno
Published: December 2019

Bispecific antibody (bsAb) applications have exponentially expanded with the advent of molecular engineering strategies that have addressed many of the initial challenges, including improper light chain pairing, heterodimer purity, aggregation, and pharmacokinetics. However, the lack of high-throughput methods for the generation of monovalent bsAbs has resulted in a bottleneck that has hampered their therapeutic evaluation, as current technologies can be cost-prohibitive and impractical. To address this issue, we incorporated single-matched point mutations in the CH3 domain to recapitulate the physiological process of human IgG4 Fab-arm exchange to generate monovalent bsAbs. Furthermore, we utilized the substitutions H435R and Y436F in the CH3 domain of IgG1, which incorporates residues from human IgG3, thus ablating protein A binding. By exploiting this combination of mutations and optimizing the reduction and reoxidation conditions for Fab arm exchange, highly pure monovalent bsAbs can be rapidly purified directly from combined culture media using standard protein A purification. This methodology, reported herein for the first time, allows for the high-throughput generation of monovalent bsAbs, thus increasing the capacity for evaluating monovalent bsAb iterations for therapeutic potential.
Curcumin-In-Deformable Liposomes-In-Chitosan-Hydrogel as a Novel Wound Dressing
Authors: Ternullo Selenia , Schulte Werning Laura Victoria , Holsæter Ann Mari , Škalko-Basnet Nataša
Published: December 2019

A liposomes-in-hydrogel system as an advanced wound dressing for dermal delivery of curcumin was proposed for improved chronic wound therapy. Curcumin, a multitargeting poorly soluble active substance with known beneficial properties for improved wound healing, was incorporated in deformable liposomes to overcome its poor solubility. Chitosan hydrogel served as a vehicle providing superior wound healing properties. The novel system should assure sustained skin delivery of curcumin, and increase its retention at the skin site, utilizing both curcumin and chitosan to improve the therapy outcome. To optimize the properties of the formulation and determine the effect of the liposomal charge on the hydrogel properties, curcumin-containing deformable liposomes (DLs) with neutral (NDLs), cationic (CDLs), and anionic (ADLs) surface properties were incorporated in chitosan hydrogel. The charged DLs affected the hydrogel’s hardness, cohesiveness, and adhesiveness. Importantly, the incorporation of DLs, regardless of their surface charge, in chitosan hydrogel did not decrease the system’s bioadhesion to human skin. Stability testing revealed that the incorporation of CDLs in hydrogel preserved hydrogel´s bioadhesiveness to a higher degree than both NDLs and ADLs. In addition, CDLs-in-hydrogel enabled the most sustained skin penetration of curcumin. The proposed formulation should be further evaluated in a chronic wound model.
Stabilisation and Growth of Metastable Form II of Fluconazole in Amorphous Solid Dispersions
Authors: Nowak Maciej , Gajda Maciej , Baranowski Przemysław , Szymczyk Patrycja , Karolewicz Bożena , Nartowski Karol P.
Published: December 2019

The crystallisation of metastable drug polymorphs in polymer matrices has been reported as a successful approach to enhance the solubility of poorly water-soluble drug molecules. This can be achieved using different polymers, drug to polymer ratios and formulation techniques enabling the formation of stable nuclei and subsequent growth of new or metastable drug polymorphs. In this work we elucidated the polymorphism behaviour of a model compound fluconazole (FLU) embedded in solid dispersions with amorphous Soluplus® (SOL) obtained using spray drying and fusion methods. The effect of humidity on the stability of FLU in the obtained dispersions was also evaluated. FLU at a drug content below 40 wt. % stayed amorphous in the dispersions prepared using the fusion method and crystallised exclusively into metastable form II at a drug content above 40 wt. % and 70% relative humidity (RH) conditions. In contrast, a mixture of forms I, II and hydrate of FLU was detected in the spray dried formulations after 14 days of storage at 40 °C/40% RH, with preferential growth of thermodynamically stable form I of FLU. This study highlights the importance of preparation techniques and the drug:polymer ratio in the formulation of amorphous solid dispersions and provides further understanding of the complex crystallisation behaviour of amorphous pharmaceuticals encapsulated in the polymer matrixes.
Spray Dried Formulations for Inhalation—Meaningful Characterisation of Powder Properties
Authors: Jüptner Angelika , Scherließ Regina
Published: December 2019

Spray drying as a particle engineering technique is of increasing interest in the field of inhalation and is already being utilised e.g., for the PulmoSphereTM products. As spray dried particles tend to agglomerate and are mechanically instable, low dose filling processes can be difficult. This study correlates powder flowability tests of spray dried formulations with filling processes with drum and dosator systems. Four pulmonary and four nasal powders with different characteristics in terms of shape, composition, and surface polarity were prepared and characterised for powder flowability according to Ph. Eur. and by powder rheometry. All formulations were filled with a manual drum TT and a dosator system. The classical flowability tests according to the Ph. Eur. showed a bad flow behaviour for hydrophilic pulmonary powders (x50 ~ 3 µm), whereas hydrophobic pulmonary particles and nasal particles (x50 ~ 25 µm) showed a better flowing behaviour. Powder rheometry supports this finding but can better differentiate flow behaviours.
Hybrid Integrated Semiconductor Lasers with Silicon Nitride Feedback Circuits
Authors: Boller Klaus-J. , van Rees Albert , Fan Youwen , Mak Jesse , Lammerink Rob , Franken Cornelis , van der Slot Peter , Marpaung David , Fallnich Carsten , Epping Jörn , Oldenbeuving Ruud , Geskus Dimitri , Dekker Ronald , Visscher Ilka , Grootjans Robert , Roeloffzen Chris , Hoekman Marcel , Klein Edwin , Leinse Arne , Heideman René
Published: December 2019

Hybrid integrated semiconductor laser sources offering extremely narrow spectral linewidth, as well as compatibility for embedding into integrated photonic circuits, are of high importance for a wide range of applications. We present an overview on our recently developed hybrid-integrated diode lasers with feedback from low-loss silicon nitride (Si 3 N 4 in SiO 2 ) circuits, to provide sub-100-Hz-level intrinsic linewidths, up to 120 nm spectral coverage around a 1.55 μ m wavelength, and an output power above 100 mW. We show dual-wavelength operation, dual-gain operation, laser frequency comb generation, and present work towards realizing a visible-light hybrid integrated diode laser.
New EST-SSR Markers for Individual Genotyping of Opium Poppy Cultivars (Papaver somniferum L.)
Authors: Vašek Jakub , Čílová Daniela , Melounová Martina , Svoboda Pavel , Vejl Pavel , Štikarová Radka , Vostrý Luboš , Kuchtová Perla , Ovesná Jaroslava
Published: December 2019

High-quality simple sequence repeat (SSR) markers are invaluable tools for revealing genetic variability which could be utilized for many purposes, such as breeding new varieties or the identifying current ones, among other applications. Based on the analysis of 3.7 million EST sequences and 15 genomic sequences from bacterial artificial chromosome (BAC) libraries, 200 trinucleotide genic (EST)-SSR and three genomic (gSSR) markers were tested, where 17 of them fulfilled all criteria for quality markers. Moreover, the reproducibility of these new markers was verified by two genetics laboratories, with a mean error rate per allele and per locus equal to 0.17%. These markers were tested on 38 accessions of Papaver somniferum and nine accessions of another five species of the Papaver and Argemone genera. In total, 118 alleles were detected for all accessions (median = 7; three to ten alleles per locus) and 88 alleles (median = 5; three to nine alleles per locus) within P. somniferum alone. Multivariate methods and identity analysis revealed high resolution capabilities of the new markers, where all but three pair accessions (41 out of 47) had a unique profile and opium poppy was distinguished from other species.
Analysis of Cultivar-Specific Variability in Size-Related Leaf Traits and Modeling of Single Leaf Area in Three Medicinal and Aromatic Plants: Ocimum basilicum L., Mentha Spp., and Salvia Spp.
Authors: Teobaldelli Maurizio , Basile Boris , Giuffrida Francesco , Romano Daniela , Toscano Stefania , Leonardi Cherubino , Rivera Carlos Mario , Colla Giuseppe , Rouphael Youssef
Published: December 2019

In this study, five allometric models were used to estimate the single leaf area of three well-known medicinal and aromatic plants (MAPs) species, namely basil (Ocimum basilicum L.), mint (Mentha spp.), and sage (Salvia spp.). MAPs world production is expected to rise up to 5 trillion US$ by 2050 and, therefore, there is a high interest in developing research related to this horticultural sector. Calibration of the models was obtained separately for three selected species by analyzing (a) the cultivar variability—i.e., 5 cultivars of basil (1094 leaves), 4 of mint (901 leaves), and 5 of sage (1103 leaves)—in the main two traits related to leaf size (leaf length, L, and leaf width, W) and (b) the relationship between these traits and single leaf area (LA). Validation of the chosen models was obtained for each species using an independent dataset, i.e., 487, 441, and 418 leaves, respectively, for basil (cv. ‘Lettuce Leaf’), mint (cv. ‘Comune’), and sage (cv. ‘Comune’). Model calibration based on fast-track methodologies, such as those using one measured parameter (one-regressor models: L, W, L2, and W2) or on more accurate two-regressors models (L × W), allowed to achieve different levels of accuracy. This approach highlighted the importance of considering intra-specific variability before applying any models to a certain cultivar to predict single LA. Eventually, during the validation phase, although modeling of single LA based on W2 showed a good fitting (R2basil = 0.948; R2mint = 0.963; R2sage = 0.925), the distribution of the residuals was always unsatisfactory. On the other hand, two-regressor models (based on the product L × W) provided the best fitting and accuracy for basil (R2 = 0.992; RMSE = 0.327 cm2), mint (R2 = 0.998; RMSE = 0.222 cm2), and sage (R2 = 0.998; RMSE = 0.426 cm2).
Copy Number Variation of Transposable Elements in Thinopyrum intermedium and Its Diploid Relative Species
Authors: Divashuk Mikhail G. , Karlov Gennady I. , Kroupin Pavel Yu.
Published: December 2019

Diploid and polyploid wild species of Triticeae have complex relationships, and the understanding of their evolution and speciation could help to increase the usability of them in wheat breeding as a source of genetic diversity. The diploid species Pseudoroegneria spicata (St), Thinopyrum bessarabicum (Jb), Dasypyrum villosum (V) derived from a hypothetical common ancestor are considered to be possible subgenome donors in hexaploid species Th. intermedium (JrJvsSt, where indices r, v, and s stand for the partial relation to the genomes of Secale, Dasypyrum, and Pseudoroegneria, respectively). We quantified 10 families of transposable elements (TEs) in P. spicata, Th. bessarabicum, D. villosum (per one genome), and Th. intermedium (per one average subgenome) using the quantitative real time PCR assay and compared their abundance within the studied genomes as well as between them. Sabrina was the most abundant among all studied elements in P. spicata, D. villosum, and Th. intermedium, and among Ty3/Gypsy elements in all studied species. Among Ty1/Copia elements, Angela-A and WIS-A showed the highest and close abundance with the exception of D. villosum, and comprised the majority of all studied elements in Th. bessarabicum. Sabrina, BAGY2, and Angela-A showed similar abundance among diploids and in Th. intermedium hexaploid; Latidu and Barbara demonstrated sharp differences between diploid genomes. The relationships between genomes of Triticeae species based on the studied TE abundance and the role of TEs in speciation and polyploidization in the light of the current phylogenetic models is discussed.
Expression and Ion Transport Activity of Rice OsHKT1;1 Variants
Authors: Imran Shahin , Horie Tomoaki , Katsuhara Maki
Published: December 2019

OsHKT1;1 in rice, belongs to the high-affinity K+ Transporter family, has been found to be involved in salt tolerance. OsHKT1;1 in japonica rice (Nipponbare) produces mRNA variants, but their functions remain elusive. In salt tolerant rice, Pokkali, eight OsHKT1;1 variants (V1-V8) were identified in addition to the full-length OsHKT1;1 (FL) cDNA. Absolute quantification by qPCR revealed that accumulation of OsHKT1;1-FL mRNA is minor in contrast to that of OsHKT1;1-V1, -V2, -V4, and -V7 mRNAs, all of which are predominant in shoots, while only V1 and V7 mRNAs are predominant in roots. Two electrode voltage clamp (TEVC) experiments using Xenopus laevis oocytes revealed that oocytes-expressing OsHKT1;1-FL from Pokkali exhibited inward-rectified currents in the presence of 96 mM Na+ as reported previously. Further TEVC analyses indicated that six of eight OsHKT1;1 variants elicited currents in a Na+ or a K+ bath solution. OsHKT1;1-V6 exhibited a similar inward rectification to the FL protein. Contrastingly, however, the rests mediated bidirectional currents in both Na+ and K+ bath solutions. These data suggest possibilities that novel mechanisms regulating the transport activity of OsHKT1;1 might exist, and that OsHKT1;1 variants might also carry out distinct physiological roles either independently or in combination with OsHKT1;1-FL.
UHPLC-Q-TOF/MS-Based Metabolomics Approach Reveals the Antifungal Potential of Pinocembroside against Citrus Green Mold Phytopathogen
Authors: Chen Chuying , Cai Nan , Chen Jinyin , Wan Chunpeng
Published: December 2019

Pinocembroside (PiCB) isolated from Ficus hirta Vahl. fruit was studied herein with the aim to find the potential mechanism for significant inhibition of growth of Penicillium digitatum, a causative pathogen of citrus green mold disease. PiCB substantially inhibited mycelial growth of P. digitatum, with the observed half maximal effective concentration (EC50), minimum inhibitory concentration (MIC), and minimum fungicidal concentration (MFC) of 120.3, 200, and 400 mg/L, respectively. Moreover, PiCB altered hyphal morphology and cellular morphology by breaking and shrinking of mycelia, decomposing cell walls, cytoplasmic inclusions. In addition to, a non-targeted metabolomics analysis by UHPLC-Q-TOF/MS was also performed, which revealed that PiCB treatment notably disrupted the metabolisms of amino acids, lipids, fatty acids, TCA, and ribonucleic acids, thereby contributing to membrane peroxidation. Current findings provide a new perception into the antifungal mechanism of PiCB treatment in inhibiting P. digitatum growth through membrane peroxidation.
Plant-Unique cis/trans Isomerism of Long-Chain Base Unsaturation is Selectively Required for Aluminum Tolerance Resulting from Glucosylceramide-Dependent Plasma Membrane Fluidity
Authors: Sato Masaya , Nagano Minoru , Jin Song , Miyagi Atsuko , Yamaguchi Masatoshi , Kawai-Yamada Maki , Ishikawa Toshiki
Published: December 2019

Cis/trans isomerism of the Δ8 unsaturation of long-chain base (LCB) is found only in plant sphingolipids. This unique geometry is generated by sphingolipid LCB Δ8 desaturase SLD which produces both isomers at various ratios, resulting in diverse cis/trans ratios in plants. However, the biological significance of this isomeric diversity remains controversial. Here, we show that the plant-specific cis unsaturation of LCB selectively contributes to glucosylceramide (GlcCer)-dependent tolerance to aluminum toxicity. We established three transgenic rice lines with altered LCB unsaturation profiles. Overexpression of SLD from rice (OsSLD-OX), which preferentially exhibits cis-activity, or Arabidopsis (AtSLD-OX), showing preference for trans-activity, facilitated Δ8 unsaturation in different manners: a slight increase of cis-unsaturated glycosylinositolphosphoceramide (GIPC) in OsSLD-OX, and a drastic increase of trans-unsaturated GlcCer and GIPC in AtSLD-OX. Disruption of LCB Δ4 desaturase (des) significantly decreased the content of GlcCer. Fluorescence imaging analysis revealed that OsSLD-OX and AtSLD-OX showed increased plasma membrane fluidity, whereas des had less fluidity, demonstrating that the isomers universally contributed to increasing membrane fluidity. However, the results of a hydroponic assay showed decreased aluminum tolerance in AtSLD-OX and des compared to OsSLD-OX and the control plants, which did not correlate with membrane fluidity. These results suggest that cis-unsaturated GlcCer, not GIPC, selectively serves to maintain the membrane fluidity specifically associated with aluminum tolerance.
Transcriptional Regulation in Rocket Leaves as Affected by Salinity
Authors: Franzoni Giulia , Cocetta Giacomo , Trivellini Alice , Ferrante Antonio
Published: December 2019

Salinity is one of the major abiotic stress causing yield losses and decreasing product quality. The beneficial effects of biostimulant products to enhance plant tolerance to abiotic stresses have been reported in several crops, but their mode of action is poorly understood. This work aims to better understand the effect of salt stress on wild rocket treated with a borage extract. The expression of some of the transcription factors (TFs) typically involved in salt stress response was studied within a 24 h period. Physiological parameters such as chlorophyll, chlorophyll a fluorescence, carotenoids, phenols, and anthocyanin were analyzed. Results obtained showed that salt stress induced a general increase in the expression levels of almost all TFs studied, whereas the treatment with the plant-base extract only induced an increase at specific time points. Moreover, the approach adopted allowed indagating the change in gene expression during time. Different pathways such as sugars metabolism, cuticular wax biosynthesis, and brassinosteroids signaling took part in plant responses.
Synthesis of Ultrahigh Molecular Weight Polymers Containing Reactive Functionality with Low PDIs by Polymerizations of Long-Chain α-Olefins in the Presence of Their Nonconjugated Dienes by Cp*TiMe2(O-2,6-iPr2C6H3)–Borate Catalyst
Authors: Nomura Kotohiro , Pengoubol Sarntamon , Apisuk Wannida
Published: December 2019

Copolymerizations of 1-decene (DC) with 1,9-decadiene (DCD), 1-dodecene (DD) with 1,11-dodecadiene (DDD), and 1-tetradecene (TD) with 1,13-tetradecadiene (TDD), using Cp*TiMe2(O-2,6-iPr2C6H3) (1)–[Ph3C][B(C6F5)4] (borate) catalyst in the presence of AliBu3/Al(n-C8H17)3 proceeded in a quasi-living manner in n-hexane at −30 to −50 °C, affording ultrahigh molecular weight (UHMW) copolymers containing terminal olefinic double bonds in the side chain with rather low PDI (Mw/Mn) values. In the DC/DCD copolymerization, the resultant copolymer prepared at −40 °C possessed UHMW (Mn = 1.40 × 106 after 45 min) with low PDI (Mw/Mn = 1.39); both the activity and the PDI value decreased at low polymerization temperature (Mn = 5.38 × 105, Mw/Mn = 1.18, after 120 min at −50 °C). UHMW poly(TD-co-TDD) was also obtained in the copolymerization at −30 °C (Mn = 9.12 × 105, Mw/Mn = 1.51, after 120 min), using this catalyst.