bluuebunny/crossref_metadata_2025 huggingface dataset:
Lipidomic Analysis of Cells and Extracellular Vesicles from High- and Low-Metastatic Triple-Negative Breast Cancer
Authors: Nishida-Aoki Nao , Izumi Yoshihiro , Takeda Hiroaki , Takahashi Masatomo , Ochiya Takahiro , Bamba Takeshi
Published: February 2020

Extracellular vesicles (EVs) are lipid bilayer nanovesicles secreted from almost all cells including cancer. Cancer-derived EVs contribute to cancer progression and malignancy via educating the surrounding normal cells. In breast cancer, epidemiological and experimental observations indicated that lipids are associated with cancer malignancy. However, lipid compositions of breast cancer EVs and their contributions to cancer progression are unexplored. In this study, we performed a widely targeted quantitative lipidomic analysis in cells and EVs derived from high- and low-metastatic triple-negative breast cancer cell lines, using supercritical fluid chromatography fast-scanning triple-quadrupole mass spectrometry. We demonstrated the differential lipid compositions between EVs and cells of their origin, and between high- and low-metastatic cell lines. Further, we demonstrated EVs from highly metastatic breast cancer accumulated unsaturated diacylglycerols (DGs) compared with EVs from lower-metastatic cells, without increasing the amount in cells. The EVs enriched with DGs could activate the protein kinase D signaling pathway in endothelial cells, which can lead to stimulated angiogenesis. Our results indicate that lipids are selectively loaded into breast cancer EVs to support tumor progression.
The Determinants of the Human Milk Metabolome and Its Role in Infant Health
Authors: Ojo-Okunola Anna , Cacciatore Stefano , Nicol Mark P. , du Toit Elloise
Published: February 2020

Human milk is needed for optimal growth as it satisfies both the nutritional and biological needs of an infant. The established relationship between breastfeeding and an infant’s health is attributable to the nutritional and non-nutritional, functional components of human milk including metabolites such as the lipids, amino acids, biogenic amines and carbohydrates. These components have diverse roles, including protecting the infant against infections and guiding the development of the infant’s immature immune system. In this review, we provide an in-depth and updated insight into the immune modulatory and anti-infective role of human milk metabolites and their effects on infant health and development. We also review the literature on potential determinants of the human milk metabolome, including maternal infectious diseases such as human immunodeficiency virus and mastitis.
Optical Frequency Combs in Quadratically Nonlinear Resonators
Authors: Ricciardi Iolanda , Mosca Simona , Parisi Maria , Leo François , Hansson Tobias , Erkintalo Miro , Maddaloni Pasquale , De Natale Paolo , Wabnitz Stefan , De Rosa Maurizio
Published: February 2020

Optical frequency combs are one of the most remarkable inventions in recent decades. Originally conceived as the spectral counterpart of the train of short pulses emitted by mode-locked lasers, frequency combs have also been subsequently generated in continuously pumped microresonators, through third-order parametric processes. Quite recently, direct generation of optical frequency combs has been demonstrated in continuous-wave laser-pumped optical resonators with a second-order nonlinear medium inside. Here, we present a concise introduction to such quadratic combs and the physical mechanism that underlies their formation. We mainly review our recent experimental and theoretical work on formation and dynamics of quadratic frequency combs. We experimentally demonstrated comb generation in two configurations: a cavity for second harmonic generation, where combs are generated both around the pump frequency and its second harmonic and a degenerate optical parametric oscillator, where combs are generated around the pump frequency and its subharmonic. The experiments have been supported by a thorough theoretical analysis, aimed at modelling the dynamics of quadratic combs, both in frequency and time domains, providing useful insights into the physics of this new class of optical frequency comb synthesizers. Quadratic combs establish a new class of efficient frequency comb synthesizers, with unique features, which could enable straightforward access to new spectral regions and stimulate novel applications.
Magnetically Driven Bionic Millirobots with a Low-Delay Automated Actuation System for Bioparticles Manipulation
Authors: Bai Xue , Chen Dixiao , Zhang Wei , Ossian Heulin , Chen Yuanyuan , Feng Yanmin , Feng Lin , Arai Fumihito
Published: February 2020

This paper presents a semi-automatic actuation system which can achieve bio-particles tracking, transportation, and high-precision motion control of robots in a microfluidic chip. This system is mainly applied in magnetically driven robots. An innovative manta ray-like robot was designed to increase stability of robots in a non-contaminated manipulation environment. A multilayer piezo actuator was applied to generate high-frequency vibration to decrease the friction between robots and the glass substrate. We also set up a user-friendly GUI (Graphical User Interface) and realized robot tracking and predetermined trajectory motion through excellent algorithms using Python and C++. In biotechnology, precise transportation of cells is used for the enucleation, microinjection, and investigation of the characteristics of a single cell. Being optimized, the parameters of the robot can effectively reach 10 µm in actuation precision and a maximum actuation speed of 200 mm/s.
Surface Response Based Modeling of Liposome Characteristics in a Periodic Disturbance Mixer
Authors: López Rubén R. , Ocampo Ixchel , Sánchez Luz-María , Alazzam Anas , Bergeron Karl-F. , Camacho-León Sergio , Mounier Catherine , Stiharu Ion , Nerguizian Vahé
Published: February 2020

Liposomes nanoparticles (LNPs) are vesicles that encapsulate drugs, genes, and imaging labels for advanced delivery applications. Control and tuning liposome physicochemical characteristics such as size, size distribution, and zeta potential are crucial for their functionality. Liposome production using micromixers has shown better control over liposome characteristics compared with classical approaches. In this work, we used our own designed and fabricated Periodic Disturbance Micromixer (PDM). We used Design of Experiments (DoE) and Response Surface Methodology (RSM) to statistically model the relationship between the Total Flow Rate (TFR) and Flow Rate Ratio (FRR) and the resulting liposomes physicochemical characteristics. TFR and FRR effectively control liposome size in the range from 52 nm to 200 nm. In contrast, no significant effect was observed for the TFR on the liposomes Polydispersity Index (PDI); conversely, FRR around 2.6 was found to be a threshold between highly monodisperse and low polydispersed populations. Moreover, it was shown that the zeta potential is independent of TFR and FRR. The developed model presented on the paper enables to pre-establish the experimental conditions under which LNPs would likely be produced within a specified size range. Hence, the model utility was demonstrated by showing that LNPs were produced under such conditions.
The Research on Multi-Material 3D Vascularized Network Integrated Printing Technology
Authors: Yang Shuai , Tang Hao , Feng Chunmei , Shi Jianping , Yang Jiquan
Published: February 2020

Three-dimensional bioprinting has emerged as one of the manufacturing approaches that could potentially fabricate vascularized channels, which is helpful to culture tissues in vitro. In this paper, we report a novel approach to fabricate 3D perfusable channels by using the combination of extrusion and inkjet techniques in an integrated manufacture process. To achieve this, firstly we investigate the theoretical model to analyze influencing factors of structural dimensions of the printed parts like the printing speed, pressure, dispensing time, and voltage. In the experiment, photocurable hydrogel was printed to form a self-supporting structure with internal channel grooves. When the desired height of hydrogel was reached, the dual print-head was switched to the piezoelectric nozzle immediately, and the sacrificial material was printed by the changed nozzle on the printed hydrogel layer. Then, the extrusion nozzle was switched to print the next hydrogel layer. Once the printing of the internal construct was finished, hydrogel was extruded to wrap the entire structure, and the construct was immersed in a CaCl2 solution to crosslink. After that, the channel was formed by removing the sacrificial material. This approach can potentially provide a strategy for fabricating 3D vascularized channels and advance the development of culturing thick tissues in vitro.
Strain-Insensitive Elastic Surface Electromyographic (sEMG) Electrode for Efficient Recognition of Exercise Intensities
Authors: Tang Daxiu , Yu Zhe , He Yong , Asghar Waqas , Zheng Ya-Nan , Li Fali , Shi Changcheng , Zarei Roozbeh , Liu Yiwei , Shang Jie , Liu Xiang , Li Run-Wei
Published: February 2020

Surface electromyography (sEMG) sensors are widely used in the fields of ergonomics, sports science, and medical research. However, current sEMG sensors cannot recognize the various exercise intensities efficiently because of the strain interference, low conductivity, and poor skin-conformability of their electrodes. Here, we present a highly conductive, strain-insensitive, and low electrode–skin impedance elastic sEMG electrode, which consists of a three-layered structure (polydimethylsiloxane/galinstan + polydimethylsiloxane/silver-coated nickel + polydimethylsiloxane). The bottom layer of the electrode consists of vertically conductive magnetic particle paths, which are insensitive to stretching strain, collect sEMG charge from human skin, and finally transfer it to processing circuits via an intermediate layer. Our skin-friendly electrode exhibits high conductivity (0.237 and 1.635 mΩ·cm resistivities in transverse and longitudinal directions, respectively), low electrode–skin impedance (47.23 kΩ at 150 Hz), excellent strain-insensitivity (10% change of electrode–skin impedance within the 0–25% strain range), high fatigue resistance (>1500 cycles), and good conformability with skin. During various exercise intensities, the signal-to-noise ratio (SNR) of our electrode increased by 22.53 dB, which is 206% and 330% more than that of traditional Ag/AgCl and copper electrode, respectively. The ability of our electrode to efficiently recognize various exercise intensities confirms its great application potential for the field of sports health.
A Potent Burkholderia Endophyte against Boxwood Blight Caused by Calonectria pseudonaviculata
Authors: Kong Ping , Hong Chuanxue
Published: February 2020

Calonectria pseudonaviculata (Cps) poses an increasing threat to boxwood, a major nursery crop and iconic landscape plant worldwide. Here, we report on a potent biocontrol agent that produces small sage green (SSG) colonies on potato dextrose agar. SSG is a bacterial strain recovered from Justin Brouwers boxwood leaves with unusual response to Cps inoculation. Water-soaked symptoms developed on leaves 2 days after inoculation then disappeared a few days later. This endophyte affected several major steps of the boxwood blight disease cycle. SSG at 107 cfu/mL lysed all conidia in mixed broth culture. SSG at 108 cfu/mL reduced blight incidence by >98% when applied one day before or 3 h after boxwood were inoculated with Cps. Its control efficacy decreased with decreasing bacterial concentration to 103 cfu/mL and increasing lead time up to 20 days. When applied on diseased leaf litter under boxwood plants, SSG reduced Cps sporulation and consequently mitigated blight incidence by 90%. SSG was identified as a new member of the Burkholderia cepacia complex with distinct characters from known clinical strains. With these protective, curative, and sanitizing properties, this Burkholderia endophyte offers great promise for sustainable blight management at production and in the landscape.
A Systematic Approach to Bacterial Phylogeny Using Order Level Sampling and Identification of HGT Using Network Science
Authors: Khaledian Ehdieh , Brayton Kelly A. , Broschat Shira L.
Published: February 2020

Reconstructing and visualizing phylogenetic relationships among living organisms is a fundamental challenge because not all organisms share the same genes. As a result, the first phylogenetic visualizations employed a single gene, e.g., rRNA genes, sufficiently conserved to be present in all organisms but divergent enough to provide discrimination between groups. As more genome data became available, researchers began concatenating different combinations of genes or proteins to construct phylogenetic trees believed to be more robust because they incorporated more information. However, the genes or proteins chosen were based on ad hoc approaches. The large number of complete genome sequences available today allows the use of whole genomes to analyze relationships among organisms rather than using an ad hoc set of genes. We present a systematic approach for constructing a phylogenetic tree based on simultaneously clustering the complete proteomes of 360 bacterial species. From the homologous clusters, we identify 49 protein sequences shared by 99% of the organisms to build a tree. Of the 49 sequences, 47 have homologous sequences in both archaea and eukarya. The clusters are also used to create a network from which bacterial species with horizontally-transferred genes from other phyla are identified.
Molecular Mechanisms Underpinning Aggregation in Acidiphilium sp. C61 Isolated from Iron-Rich Pelagic Aggregates
Authors: Li Qianqian , Cooper Rebecca E. , Wegner Carl-Eric , Küsel Kirsten
Published: February 2020

Iron-rich pelagic aggregates (iron snow) are hot spots for microbial interactions. Using iron snow isolates, we previously demonstrated that the iron-oxidizer Acidithrix sp. C25 triggers Acidiphilium sp. C61 aggregation by producing the infochemical 2-phenethylamine (PEA). Here, we showed slightly enhanced aggregate formation in the presence of PEA on different Acidiphilium spp. but not other iron-snow microorganisms, including Acidocella sp. C78 and Ferrovum sp. PN-J47. Next, we sequenced the Acidiphilium sp. C61 genome to reconstruct its metabolic potential. Pangenome analyses of Acidiphilium spp. genomes revealed the core genome contained 65 gene clusters associated with aggregation, including autoaggregation, motility, and biofilm formation. Screening the Acidiphilium sp. C61 genome revealed the presence of autotransporter, flagellar, and extracellular polymeric substances (EPS) production genes. RNA-seq analyses of Acidiphilium sp. C61 incubations (+/− 10 µM PEA) indicated genes involved in energy production, respiration, and genetic processing were the most upregulated differentially expressed genes in the presence of PEA. Additionally, genes involved in flagellar basal body synthesis were highly upregulated, whereas the expression pattern of biofilm formation-related genes was inconclusive. Our data shows aggregation is a common trait among Acidiphilium spp. and PEA stimulates the central cellular metabolism, potentially advantageous in aggregates rapidly falling through the water column.
Effects of Different Yeasts on Physicochemical and Oenological Properties of Red Dragon Fruit Wine Fermented with Saccharomyces cerevisiae, Torulaspora delbrueckii and Lachancea thermotolerans
Authors: Jiang Xiaohui , Lu Yuyun , Liu Shao Quan
Published: February 2020

A new type of fruit wine made from red dragon fruit juice was produced through alcoholic fermentation (AF) with different yeasts: Saccharomyces cerevisiae EC-1118, Torulaspora delbrueckii Biodiva and Lachancea thermotolerans Concerto. Complete AF with similar fermentation rates in terms of sugar utilisation and ethanol production (8–9%, v/v) was achieved by three yeast strains. T. delbrueckii produced a significantly lower amount of glycerol and acetic acid, while L. thermotolerans produced more lactic and succinic acids. In addition, the two non-Saccharomyces strains were more efficient in proline utilisation. For volatile compounds, S. cerevisiae produced the highest amounts of esters, while T. delbrueckii produced more higher alcohols, isoamyl acetate and terpenes. On the other hand, AF caused significant degradation of betacyanin pigments and total phenolic compounds. Nevertheless, better retention of antioxidant activity and colour stability was found in L. thermotolerans and T. delbrueckii fermented wines than that of S. cerevisiae. This study suggested that it is feasible to use pure non-Saccharomyces yeast to produce red dragon fruit wine for commercialization.
Comparative Studies on the Polymorphism and Copy Number Variation of mtSSU rDNA in Ciliates (Protista, Ciliophora): Implications for Phylogenetic, Environmental, and Ecological Research
Authors: Wang Yurui , Jiang Yaohan , Liu Yongqiang , Li Yuan , Katz Laura A. , Gao Feng , Yan Ying
Published: February 2020

While nuclear small subunit ribosomal DNA (nSSU rDNA) is the most commonly-used gene marker in studying phylogeny, ecology, abundance, and biodiversity of microbial eukaryotes, mitochondrial small subunit ribosomal DNA (mtSSU rDNA) provides an alternative. Recently, both copy number variation and sequence variation of nSSU rDNA have been demonstrated for diverse organisms, which can contribute to misinterpretation of microbiome data. Given this, we explore patterns for mtSSU rDNA among 13 selected ciliates (representing five classes), a major component of microbial eukaryotes, estimating copy number and sequence variation and comparing to that of nSSU rDNA. Our study reveals: (1) mtSSU rDNA copy number variation is substantially lower than that for nSSU rDNA; (2) mtSSU rDNA copy number ranges from 1.0 × 104 to 8.1 × 105; (3) a most common sequence of mtSSU rDNA is also found in each cell; (4) the sequence variation of mtSSU rDNA are mainly indels in poly A/T regions, and only half of species have sequence variation, which is fewer than that for nSSU rDNA; and (5) the polymorphisms between haplotypes of mtSSU rDNA would not influence the phylogenetic topology. Together, these data provide more insights into mtSSU rDNA as a powerful marker especially for microbial ecology studies.
Biological Control of Fruit Rot and Anthracnose of Postharvest Mango by Antagonistic Yeasts from Economic Crops Leaves
Authors: Konsue Wilasinee , Dethoup Tida , Limtong Savitree
Published: February 2020

To select antagonistic yeasts for the control of fruit rot caused by Lasiodiplodia theobromae and anthracnose caused by Colletotrichum gloeosporioides in postharvest mango fruit, 307 yeast strains isolated from plant leaves were evaluated for their antagonistic activities against these two fungal pathogens in vitro. Torulaspora indica DMKU-RP31, T. indica DMKU-RP35 and Pseudozyma hubeiensis YE-21 were found to inhibit the growth of L. theobromae whereas only Papiliotrema aspenensis DMKU-SP67 inhibited the growth of C. gloeosporioides. Antagonistic mechanisms of these four antagonistic yeasts in vitro consisted of the production of antifungal volatile organic compounds (VOCs), biofilm formation and siderophore production. T. indica DMKU-RP35 was the most effective strain in controlling fruit rot on postharvest mango fruits. Its action was comparable to that of the fungicide, benomyl, reducing the disease severity by 82.4%, whereas benomyl revealed 87.5% reduction. P. aspenensis DMKU-SP67 reduced anthracnose severity by 94.1%, which was comparable to that of using benomyl (93.9%). The antifungal VOCs produced by these yeast strains also reduced the severity of these diseases on postharvest mango fruits but at lower rates than using yeast cells. Therefore, these antagonistic yeasts have the potential for use as biological control agents for the control of fruit rot and anthracnose diseases.
Identification of Rhizospheric Actinomycete Streptomyces lavendulae SPS-33 and the Inhibitory Effect of its Volatile Organic Compounds against Ceratocystis fimbriata in Postharvest Sweet Potato (Ipomoea batatas (L.) Lam.)
Authors: Li Xuewei , Li Beibei , Cai Shurui , Zhang Yu , Xu Mingjie , Zhang Chunmei , Yuan Bo , Xing Ke , Qin Sheng
Published: February 2020

Black spot disease, which is caused by the pathogenic fungal Ceratocystis fimbriata, seriously affects the production of sweet potato and its quality during postharvest storage. In this study, the preliminary identification of the rhizosphere actinomycete strain SPS-33, and its antifungal activity of volatiles in vitro and in vivo was investigated. Based on morphological identification and phylogenetic analysis of the 16S rRNA gene sequence, strain SPS-33 was identified as Streptomyces lavendulae. Volatile organic compounds (VOCs) emitted by SPS-33 inhibited mycelial growth and sporulation of C. fimbriata in vitro and also induced a series of observable hyphae morphological changes. In an in vivo pathogenicity assay, exposure to SPS-33 significantly decreased the lesion diameter and water loss rate in sweet potato tuberous roots (TRs) inoculated with C. fimbriata. It increased the antioxidant enzymes’ activities of peroxidase, catalase, and superoxide dismutase as well as decreased malondialdehyde and increased total soluble sugar. In the VOC profile of SPS-33 detected by a headspace solid-phase micro extraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS), heptadecane, tetradecane, and 3-methyl-1-butanol were the most abundant compounds. 2-Methyl-1-butanol, 3-methyl-1-butanol, pyridine, and phenylethyl alcohol showed strong antifungal effects against C. fimbriata. These findings suggest that VOCs from S. lavendulae SPS-33 have the potential for pathogen C. fimbriata control in sweet potato postharvest storage by fumigant action.
X-ray Laue Microdiffraction and Raman Spectroscopic Investigation of Natural Silicon and Moissanite
Authors: Stan Camelia Veronica , O’Bannon Earl Francis , Mukhin Pavel , Tamura Nobumichi , Dobrzhinetskaya Larissa
Published: February 2020

Moissanite, SiC, is an uncommon accessory mineral that forms under low oxygen fugacity. Here, we analyze natural SiC from a Miocene tuff-sandstone using synchrotron Laue microdiffraction and Raman spectroscopy, in order to better understand the SiC phases and formation physics. The studied crystals of SiC consist of 4H- and 6H-SiC domains, formed from either, continuous growth or, in one case, intergrown, together with native Si. The native Si is polycrystalline, with a large crystal size relative to the analytical beam dimensions (>1–2 μm). We find that the intergrown region shows low distortion or dislocation density in SiC, but these features are comparatively high in Si. The distortion/deformation observed in Si may have been caused by a mismatch in the coefficients of thermal expansion of the two materials. Raman spectroscopic measurements are discussed in combination with our Laue microdiffraction results. Our results suggest that these SiC grains likely grew from an igneous melt.
Geochronology of Uraninite Revisited
Authors: Corcoran Loretta , Simonetti Antonio
Published: February 2020

Identification of uraninite provenance for the purpose of nuclear forensics requires a multifaceted approach. Various geochemical signatures, such as chondrite normalized rare earth element patterns, help identify and limit the potential sources of uraninite based on the geological setting of the uranium ore mineralization. The inclusion of accurate age determinations to discriminate geochemical signatures for natural uranium ores may help to potentially restrict geographical areas for provenance consideration. Determining a robust age for uraninite formation is somewhat difficult, due to well known, inherent difficulties associated with open system behavior that involve either uranium and/or lead loss or gain. However, open system behavior should not perturb their Pb isotopic compositions to the same degree as Pb isotopes should not fractionate during alteration processes. Here, a suite of pristine and altered samples of uraninite was examined for their Pb isotope compositions, and these yielded geologically meaningful secondary Pb–Pb isochron ages. The degree of alteration within individual uraninite samples, which is extremely variable, does not appear to affect the calculated ages. The approach adopted here yields insightful age information, and hence, is of great value for source attribution in forensic analyses of raw nuclear materials.
Enhanced Photo-Assisted Acetone Gas Sensor and Efficient Photocatalytic Degradation Using Fe-Doped Hexagonal and Monoclinic WO3 Phase−Junction
Authors: Wang Ji-Chao , Shi Weina , Sun Xue-Qin , Wu Fang-Yan , Li Yu , Hou Yuxia
Published: February 2020

The development of WO3-based gas sensors for analysis of acetone in exhaled breath is significant for noninvasive diagnosis of diabetes. A series of Fe-doped hexagonal and monoclinic WO3 phase−junction (Fe−h/m−WO3) sensors were synthesized by the hydrothermal calcination method, and the influences of operating temperature and light irradiation on the response were studied. Under light emitting diode (LED) illumination, Fe−h/m−WO3 exhibited higher responses to acetone than those of the undoped WO3-based sensors at an operating temperature of 260 °C with 90% relative humidity, and good linearity between response and acetone concentration (0.5 to 2.5 ppm) was achieved under the 90% relative humidity condition. Meanwhile, the optimal Fe−h/m−WO3 sensor exhibited high selectivity and stability for a duration of three months. The excellent sensing performance of Fe−h/m−WO3 was attributed to the formation of phase−junction and Fe doping, and these were beneficial for the separation of photon−generated carriers and oxygen adsorption on the WO3 surface, promoting the generation of superoxide radicals, which was demonstrated by electron paramagnetic resonance and photocurrent tests. Additionally, the Fe−doped WO3 phase−junction sample also showed good photocatalytic performance for rhodamine B degradation. This study may provide some insights into rational design of new types of gas sensors and offer an alternative for noninvasive diagnosis of diabetes.
The Effect of Sheep and Cow Milk Supplementation of a Low Calcium Diet on the Distribution of Macro and Trace Minerals in the Organs of Weanling Rats
Authors: Burrow Keegan , Young Wayne , McConnell Michelle , Carne Alan , Barr David , Reid Malcolm , Bekhit Alaa El-Din
Published: February 2020

The aim of this study was to investigate the effect of either sheep or cow milk supplementation to a low calcium and phosphorus diet on growth and organ mineral distribution in weanling rats. Rats were fed diets consisting of either a control chow, a 50% reduced calcium and phosphorous chow (low Ca/P), low Ca/P and sheep milk, or low Ca/P and cow milk diet for 28 days. Food intake of the rats, the growth rate of the rats, and the concentrations of minerals in the soft organs and serum were determined. Rats fed the low Ca/P diet alone had lower weight gain than rats consuming either of the milk-supplemented diets (p < 0.05). Both sheep milk and cow milk supplementation overcame the effects of consuming a diet restricted in calcium and phosphorus but the sheep milk was effective at a significantly lower level of milk intake (p < 0.05). Significant differences (p < 0.05) in essential and trace mineral concentrations due to milk type were observed in the kidney, spleen, and liver. For non-essential minerals, significant differences (p < 0.05), related to diet, were observed in all organs for arsenic, cesium, rubidium, and strontium concentrations.
Development and Validation of an in-line API Quantification Method Using AQbD Principles Based on UV-Vis Spectroscopy to Monitor and Optimise Continuous Hot Melt Extrusion Process
Authors: Almeida Juan , Bezerra Mariana , Markl Daniel , Berghaus Andreas , Borman Phil , Schlindwein Walkiria
Published: February 2020

A key principle of developing a new medicine is that quality should be built in, with a thorough understanding of the product and the manufacturing process supported by appropriate process controls. Quality by design principles that have been established for the development of drug products/substances can equally be applied to the development of analytical procedures. This paper presents the development and validation of a quantitative method to predict the concentration of piroxicam in Kollidon® VA 64 during hot melt extrusion using analytical quality by design principles. An analytical target profile was established for the piroxicam content and a novel in-line analytical procedure was developed using predictive models based on UV-Vis absorbance spectra collected during hot melt extrusion. Risks that impact the ability of the analytical procedure to measure piroxicam consistently were assessed using failure mode and effect analysis. The critical analytical attributes measured were colour (L* lightness, b* yellow to blue colour parameters—in-process critical quality attributes) that are linked to the ability to measure the API content and transmittance. The method validation was based on the accuracy profile strategy and ICH Q2(R1) validation criteria. The accuracy profile obtained with two validation sets showed that the 95% β-expectation tolerance limits for all piroxicam concentration levels analysed were within the combined trueness and precision acceptance limits set at ±5%. The method robustness was tested by evaluating the effects of screw speed (150–250 rpm) and feed rate (5–9 g/min) on piroxicam content around 15% w/w. In-line UV-Vis spectroscopy was shown to be a robust and practical PAT tool for monitoring the piroxicam content, a critical quality attribute in a pharmaceutical HME process.
Achyranthis radix Extract-Loaded Eye Drop Formulation Development and Novel Evaluation Method for Dry Eye Treatment
Authors: Kim Sung-Jin , Park Bongkyun , Huh Hyun Wook , Na Young-Guk , Kim Minki , Han Mingu , Lee Hyunmin , Pham Thi Mai Anh , Lee Hong-Ki , Lee Jae-Young , Kim Chan-Sik , Baek Jong-Suep , Cho Cheong-Weon
Published: February 2020

Recently, Achyranthis radix extract has been studied as a therapeutic agent for dry eye disease that occurs from fine dust. The aim of this study was the development of Achyranthis radix extract-loaded eye drop formulations using lubricants, generally used for artificial tear eye drops. Ecdysterone was used as a marker compound for Achyranthis radix extract and 1% Achyranthis radix extract solution contained 14.37 ± 0.04 μg/mL of ecdysterone. Before formulation studies, a new method was performed to evaluate pigmentation, which might be caused by eye drops of herbal extract. A comparative study of the water retention ability of each formulation and ability to prevent the death of conjunctival epithelial cells in dry conditions was conducted. Moreover, treatment of Achyranthis radix extract (USL) eye drop formulation exhibited a significant inhibitory effect on inflammation in a concentration-dependent manner. The long-term and accelerated stability tests showed that lubricants could contribute to the stability of herbal extracts in solution. In conclusion, hyaluronic acid showed a good effect on the development of eye drop formulation using Achyranthis radix extracts for treating dry eye disease.
pH-Sensitive Mixed Micelles Assembled from PDEAEMA-PPEGMA and PCL-PPEGMA for Doxorubicin Delivery: Experimental and DPD Simulations Study
Authors: Yang Chufen , Liu Wenyao , Xiao Jiayu , Yuan Cong , Chen Yaoxi , Guo Jianwei , Yue Hangbo , Zhu Dongyu , Lin Wenjing , Tang Shengqiu , Dong Xiaoying
Published: February 2020

To decrease critical micelle concentration (CMC), improve stability, and keep high drug-loading capacity, three pH-sensitive mixed micelles applied for anticancer drug controlled delivery were prepared by the mixture of polymers poly (N,N-diethylaminoethyl methacrylate)-b-poly(poly(ethylene glycol) methyl ether methacrylate) (PDEAEMA-PPEGMA) and polycaprolactone-b-poly (poly(ethylene glycol) methyl ether methacrylate) (PCL-PPEGMA), which were synthesized and confirmed by 1H NMR and gel permeation chromatographic (GPC). The critical micelle concentration (CMC) values of the prepared mixed micelles were low, and the micellar sizes and zeta potentials of the blank mixed micelles demonstrated good pH-responsive behavior. Combined experimental techniques with dissipative particle dynamics (DPD) simulation, the particle sizes, zeta potentials, drug loading content (LC), encapsulation efficiency (EE), aggregation morphologies, and doxorubicin (DOX) distribution of the mixed micelles were investigated, and the high DOX-loading capacity of the mixed micelles was found. Both in vitro DOX release profiles and DPD simulations of the DOX dynamics release process exhibited less leakage and good stability in neutral conditions and accelerated drug release behavior with a little initial burst in slightly acidic conditions. Cytotoxicity tests showed that the polymer PDEAEMA-PPEGMA and the blank mixed micelles had good biocompatibility, and DOX-loaded mixed micelles revealed certain cytotoxicity. These results suggest that the drug-loaded mixed micelles that consisted of the two polymers PDEAEMA-PPEGMA and PCL-PPEGMA can be new types of pH-responsive well-controlled release anticancer drug delivery mixed micelles.
Usefulness of Artificial Membrane, Strat-M®, in the Assessment of Drug Permeation from Complex Vehicles in Finite Dose Conditions
Authors: Arce Florencio Jr , Asano Narumi , See Gerard Lee , Itakura Shoko , Todo Hiroaki , Sugibayashi Kenji
Published: February 2020

The ban on the use of animals in testing cosmetic products has led to the development of animal-free in vitro methods. Strat-M® is an artificial membrane engineered to mimic human skin and is recommended as a replacement for skin. However, its usefulness in the assessment of the permeation of cosmetics in in-use conditions remains unverified. No data have been published on its comparative performance with the membrane of choice, porcine skin. The comparative permeability characteristics of Strat-M® and porcine skin were investigated using Franz diffusion cells. Caffeine (CF) and rhododendrol (RD) in complex vehicles with varying concentrations of polyols were applied as finite and infinite doses. Good rank orders of permeation from finite dose experiments were observed for RD. High correlations were observed in RD permeation between Strat-M® and porcine skin under finite and infinite dose conditions, whereas only finite dose conditions for CF were associated with good correlations. Permeation from formulations with high polyol content and residual formulations was enhanced due to the disruption of the integrity of the Strat-M® barrier. The usefulness of Strat-M® in the assessment of dermal permeation may be limited to finite dose conditions and not applicable to infinite dose conditions or formulations applied in layers.
Drug Delivery of siRNA Therapeutics
Authors: Lamberti Gaetano , Barba Anna Angela
Published: February 2020

Small interfering RNA (siRNA) is a class of nucleic acid-based drugs (NABDs) able to block gene expression by interaction with mRNA before its translation [...]
Oleogels with Birch Bark Dry Extract: Extract Saving Formulations through Gelation Enhancing Additives
Authors: Ghaffar Kashif Ahmad , Daniels Rolf
Published: February 2020

Triterpenes from the outer bark of birch have many beneficial biological and pharmacological activities. In particular, its wound healing efficacy is of paramount importance. Apart from that, particles of a birch bark dry extract aggregate into a three dimensional network when they are dispersed in lipids yielding a semi-solid oleogel. However, gel formation requires high amounts of the extract, which then acts at once as the active ingredient and the gelling agent. Infrared spectra of the respective mixtures proved that hydrogen bonds play a crucial role in the formation of the gel network. Dicarboxylic acids had almost no effect on gel strength. Monoalcohols increased the firmness of the oleogel with a decreasing effect from methanol > ethanol > butanol > octanol. All tested terminal diols increased the gel strength whereas vicinal diols affected the gel strength negatively. The effect was highly dependent on their concentration. The different effects of the diols are linked to their structure and polarity. The most pronounced enhancement of gelation was found for 1,6-hexanediol, which reduced the amount of triterpene extract (TE), which is necessary for the formation of an oleogel by a factor of 10.
Sphingomyelin-Based Nanosystems (SNs) for the Development of Anticancer miRNA Therapeutics
Authors: Nagachinta Surasa , Bouzo Belen Lopez , Vazquez-Rios Abi Judit , Lopez Rafael , Fuente Maria de la
Published: February 2020

Gene replacement therapy with oncosuppressor microRNAs (miRNAs) is a promising alternative to interfere with cancer progression. However, miRNAs are highly inefficient in a biological environment, hampering a successful translation to clinics. Nanotechnology can tackle this drawback by providing delivery systems able to efficiently deliver them to cancer cells. Thus, the objective of this work was to develop biocompatible nanosystems based on sphingomyelin (SM) for the intracellular delivery of miRNAs to colorectal cancer cells. We pursued two different approaches to select the most appropriate composition for miRNA delivery. On the one hand, we prepared sphingomyelin-based nanosystems (SNs) that incorporate the cationic lipid stearylamine (ST) to support the association of miRNA by the establishment of electrostatic interactions (SNs–ST). On the other hand, the cationic surfactant (DOTAP) was used to preform lipidic complexes with miRNA (Lpx), which were further encapsulated into SNs (SNs-Lpx). Restitution of miRNA145 levels after transfection with SNs-Lpx was related to the strongest anticancer effect in terms of tumor proliferation, colony forming, and migration capacity assays. Altogether, our results suggest that SNs have the potential for miRNA delivery to develop innovative anticancer therapies.
Assessing Performance and Engagement on a Computer-Based Education Platform for Pharmacy Practice
Authors: Grindrod Kelly , Morris Katherine , Killeen Rosemary
Published: February 2020

A computer-based education platform was developed using a theory-based approach to help Canadian pharmacy professionals adopt their full scope of practice. Data from the platform were used to identify factors that impacted user performance and engagement. A de-identified dataset included response data for 21 unique modules, including quiz responses and self-reflection questions. Outcome measures included user performance (mean quiz score) and engagement (completion rate for attempted modules). Analysis of variance (ANOVA), multivariate regression modelling, and machine learning cluster analysis were used to analyze the data. Of the 5290 users, 68% were pharmacists, 11% were technicians, 13% were pharmacy students, and 8% were pharmacy technician students. Four clusters were identified separately for pharmacists and technicians. Clusters with the higher performance and engagement tended to have more users practicing in community pharmacies while the lower performing clusters tended have more internationally trained users. In the regression modelling, pharmacists performed better than technicians and students while students were more engaged (p < 0.0001). Further, internationally trained pharmacists had slightly lower scores but similar engagement compared to domestically trained pharmacists (p < 0.0001). Users demonstrated higher performance on modules related to scope of practice than on clinical topics, and were most engaged with topics directly impacting daily practice such as influenza vaccinations and new and emerging subjects such as cannabis. The cluster analysis suggests that performance and engagement with a computer-based educational platform in pharmacy may be more related to place of practice than to personal demographic factors such as age or gender.
Side-lobe Level Reduction of an Optical Phased Array Using Amplitude and Phase Modulation of Array Elements Based on Optically Injection-Locked Semiconductor Lasers
Authors: Nguyen Anh Hang , Cho Jun-Hyung , Bae Ho-Jun , Sung Hyuk-Kee
Published: February 2020

The side-lobe level (SLL) in optical phased array (OPA) systems should be reduced to ensure their high performance. We investigate theoretically the performance of an OPA based on optically injection-locked (OIL) semiconductor lasers. The phase and amplitude of the OIL laser are modulated by controlling the injection-locking parameters to reduce the SLL as well as to achieve beam steering. We successfully achieved an SLL reduction of >16 dB when compared with the uniform amplitude profile based on the application of the Taylor window function profile to the injection-locked OPA elements. The reduced SLL and high power efficiency achieved in this study can expedite the use of OPA in real field applications, such as free-space communication, imaging, and light detection and ranging (LIDAR).
An Efficient Method for In Vitro Shoot-Tip Culture and Sporophyte Production Using Selaginella martensii Spring Sporophyte
Authors: Park Kyungtae , Jang Bo Kook , Lee Ha Min , Cho Ju Sung , Lee Cheol Hee
Published: February 2020

Selaginella martensii, an evergreen perennial fern that is native to South America and New Zealand, is named “frosty fern” because of its beautiful white-colored leaves and it is used as an ornamental plant. Efficient propagation methods for this species have not been developed. We aimed to develop an efficient propagation method for S. martensii through in vitro culture. We investigated culture conditions that are suitable for shoot-tip proliferation and growth. The optimum shoot-tip culture conditions were determined while using Murashige and Skoog (MS) medium (quarter, half, full, or double strength) and macronutrients (sucrose and two nitrogen sources) at various concentrations. In MS medium, the shoot tips formed a maximum of 6.77 nodes per explant, and each node formed two new shoot tips (i.e., 26 or 64 shoot tips). When using branching segments containing an angle meristem, the shoot-to-rhizophore formation ratio could be controlled by medium supplementation with plant-growth regulators. Sporophytes that were grown from shoot tips in vitro were acclimated in ex vitro soil conditions and successfully survived in the greenhouse. Numerous shoot tips could be obtained from in vitro-grown sporophytes and be proliferated ex vitro to produce a large number of plants. This method provides a way of shortening the time that is required for producing a large stock of S. martensii planting material.
The Effect of Infrared Drying on Color, Projected Area, Drying Time, and Total Phenolic Content of Rose (Rose electron) Petals
Authors: Selvi Kemal Çağatay , Kabutey Abraham , Gürdil Gürkan Alp Kağan , Herak David , Kurhan Şebnem , Klouček Pavel
Published: February 2020

The effects of different drying temperatures (50, 60, 70 °C) on the quality of rose (Rose electron) petals were evaluated in this study. Drying time decreased from 1680 s to 600 s with increased infrared temperature. The temperature and time were increased from 50 °C to 70 °C and 30 min to 60 min, respectively, and a decrease in the fruit color quality was observed. The projected area (PA) of rose petals was affected significantly from temperature. After the drying process, the largest PA was observed as 33.35 cm2 (50 °C, 30 min), while the smallest achieved at 70 °C, 60 min (27.96 cm2). Depending on the temperature values (50, 60, 70 °C), the average projection area of dry samples of the rose petals decreased 2.17 times compared to the projection area of fresh samples. The dried samples demonstrated an increase in the total phenolic (TP) content compared to the fresh samples. The maximum TP (44.49 mg GAE/g) was achieved at 45 min and 70 °C rose petals sample. The results concluded that infrared drying for 45 min at 70 °C could be recommended for drying rose (rosa electron) petals.
Homologous Proteins of the Manganese Transporter PAM71 Are Localized in the Golgi Apparatus and Endoplasmic Reticulum
Authors: Hoecker Natalie , Honke Anna , Frey Katharina , Leister Dario , Schneider Anja
Published: February 2020

Chloroplast manganese transporter 1 (CMT1) and photosynthesis-affected mutant 71 (PAM71) are two membrane proteins that function sequentially to mediate the passage of manganese across the chloroplast envelope and the thylakoid membrane. CMT1 and PAM71 belong to a small five-member protein family in Arabidopsis thaliana. The other three, photosynthesis-affected mutant 71 like 3 (PML3), PML4 and PML5 are not predicted to reside in chloroplast membranes. In this study, the subcellular localization of PML3:GFP, PML4:GFP and PML5:GFP was determined using transient and stable expression assays. PML3:GFP localizes to the Golgi apparatus, whereas PML4:GFP and PML5:GFP are found in the endoplasmic reticulum. We also examined patterns of PML3, PML4 and PML5 promoter activity. Although the precise expression pattern of each promoter was unique, all three genes were expressed in the leaf vasculature and in roots. Greenhouse grown single mutants pml3, pml4, pml5 and the pml4/pml5 double mutant did not exhibit growth defects, however an inspection of the root growth revealed a difference between pml3 and the other genotypes, including wild-type, in 500 µM manganese growth conditions. Strikingly, overexpression of PML3 resulted in a stunted growth phenotype. Putative functions of PML3, PML4 and PML5 are discussed in light of what is known about PAM71 and CMT1.
β-Aminobutyric Acid Pretreatment Confers Salt Stress Tolerance in Brassica napus L. by Modulating Reactive Oxygen Species Metabolism and Methylglyoxal Detoxification
Authors: Mahmud Jubayer Al , Hasanuzzaman Mirza , Khan M. Iqbal R. , Nahar Kamrun , Fujita Masayuki
Published: February 2020

Salinity is a serious environmental hazard which limits world agricultural production by adversely affecting plant physiology and biochemistry. Hence, increased tolerance against salt stress is very important. In this study, we explored the function of β-aminobutyric acid (BABA) in enhancing salt stress tolerance in rapeseed (Brassica napus L.). After pretreatment with BABA, seedlings were exposed to NaCl (100 and 150 mM) for 2 days. Salt stress increased Na content and decreased K content in shoot and root. It disrupted the antioxidant defense system by producing reactive oxygen species (ROS; H2O2 and O2•−), methylglyoxal (MG) content and causing oxidative stress. It also reduced the growth and photosynthetic pigments of seedlings but increased proline (Pro) content. However, BABA pretreatment in salt-stressed seedlings increased ascorbate (AsA) and glutathione (GSH) contents; GSH/GSSG ratio; and the activities of ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), glutathione reductase (GR), glutathione peroxidase (GPX), superoxide dismutase (SOD), catalase (CAT), glyoxalase I (Gly I), and glyoxalase II (Gly II) as well as the growth and photosynthetic pigments of plants. In addition, compared to salt stress alone, BABA increased Pro content, reduced the H2O2, MDA and MG contents, and decreased Na content in root and increased K content in shoot and root of rapeseed seedlings. Our findings suggest that BABA plays a double role in rapeseed seedlings by reducing Na uptake and enhancing stress tolerance through upregulating the antioxidant defense and glyoxalase systems.