bluuebunny/crossref_metadata_2025 huggingface dataset:
Current Stimuli-Responsive Mesoporous Silica Nanoparticles for Cancer Therapy
Authors: Moodley Thashini , Singh Moganavelli
Published: January 2021

With increasing incidence and mortality rates, cancer remains one of the most devastating global non-communicable diseases. Restricted dosages and decreased bioavailability, often results in lower therapeutic outcomes, triggering the development of resistance to conventionally used drug/gene therapeutics. The development of novel therapeutic strategies using multimodal nanotechnology to enhance specificity, increase bioavailability and biostability of therapeutics with favorable outcomes is critical. Gated vectors that respond to endogenous or exogenous stimuli, and promote targeted tumor delivery without prematurely cargo loss are ideal. Mesoporous silica nanoparticles (MSNs) are effective delivery systems for a variety of therapeutic agents in cancer therapy. MSNs possess a rigid framework and large surface area that can incorporate supramolecular constructs and varying metal species that allow for stimuli-responsive controlled release functions. Its high interior loading capacity can incorporate combination drug/gene therapeutic agents, conferring increased bioavailability and biostability of the therapeutic cargo. Significant advances in the engineering of MSNs structural and physiochemical characteristics have since seen the development of nanodevices with promising in vivo potential. In this review, current trends of multimodal MSNs being developed and their use in stimuli-responsive passive and active targeting in cancer therapy will be discussed, focusing on light, redox, pH, and temperature stimuli.
Real-World Efficacy and Safety of Fluocinolone Acetonide Implant for Diabetic Macular Edema: A Systematic Review
Authors: Kodjikian Laurent , Baillif Stephanie , Creuzot-Garcher Catherine , Delyfer Marie-Noëlle , Matonti Frédéric , Weber Michel , Mathis Thibaud
Published: January 2021

To assess real-world outcomes of fluocinolone acetonide (FAc) implant in treating diabetic macular edema (DME), a systematic literature review was conducted on PubMed in order to identify publications assessing the efficacy and safety of the FAc implant in DME in daily practice. Case reports and randomized controlled trials were excluded. Twenty-two observational real-world studies analyzing a total of 1880 eyes were included. Mean peak visual gain was +8.7 letters (11.3 months post-FAc injection) and was greater for lower baseline best corrected visual acuity (BCVA) and for more recent DME. Mean central retinal thickness (CRT) decreased 34.3% from baseline. 77.0% of the analyzed studies reported both BCVA improvement of at least five letters and a CRT decrease by 20% or more. Rescue therapy was needed more frequently when FAc was administered for chronic DME. FAc-induced ocular hypertension was reported in 20.1% of patients but only 0.6% needed surgery. Cataract extraction was performed in 43.2% of phakic patients. Adequate patient selection is essential for optimal FAc response and better safety profile. Currently positioned as second- or third-line treatment in the management algorithm, FAc implant decreases treatment burden and provides better letter gain when administered for more recent DME.
Targeting Germ Cell Tumors with the Newly Synthesized Flavanone-Derived Compound MLo1302 Efficiently Reduces Tumor Cell Viability and Induces Apoptosis and Cell Cycle Arrest
Authors: Lobo João , Cardoso Ana Rita , Miranda-Gonçalves Vera , Looijenga Leendert H. J. , Lopez Marie , Arimondo Paola B. , Henrique Rui , Jerónimo Carmen
Published: January 2021

Less toxic treatment strategies for testicular germ cell tumor (TGCT) patients are needed, as overtreatment is a concern due to the long-term side effects of platin-based chemotherapy. Although clinical benefit from classical hypomethylating agents has to date been limited, TGCTs show an abnormal DNA methylome indicating the potential of treating TGCTs with hypomethylating drugs. We tested, for the first time in TGCT cell lines, a new synthetic flavonoid compound (MLo1302) from the 3-nitroflavanone family of DNA methyltransferase (DNMT) inhibitors. We show that MLo1302 reduces cell viability (including of cisplatin resistant cell line NCCIT-R), with IC50s (inhibitory concentration 50) within the nanomolar range for NCCIT and NTERA-2 cells, and proved its cytotoxic effect. Exposure to MLo1302 reduced DNMT protein expression, similar to decitabine, and showed a partial effect in cell differentiation, reducing protein expression of pluripotency markers. RT2 profiler expression array indicated several dysregulated targets, related to activation of apoptosis, differentiation, and cell cycle arrest. We validated these data by showing increased apoptosis, increased protein expression of cleaved caspase 8 and activated caspase 2, and reduced proliferation (BrdU assay), with increase in CDKN1A and decrease in MIB-1 expression. Therefore, synthetic drugs designed to target DNA methylation in cells may uncover effective treatments for TGCT patients.
Coincubation as miR-Loading Strategy to Improve the Anti-Tumor Effect of Stem Cell-Derived EVs
Authors: Brossa Alessia , Tapparo Marta , Fonsato Valentina , Papadimitriou Elli , Delena Michela , Camussi Giovanni , Bussolati Benedetta
Published: January 2021

Extracellular vesicles are considered a novel therapeutic tool, due to their ability to transfer their cargoes to target cells. Different strategies to directly load extracellular vesicles with RNA species have been proposed. Electroporation has been used for the loading of non-active vesicles; however, the engineering of vesicles already carrying a therapeutically active cargo is still under investigation. Here, we set up a coincubation method to increase the anti-tumor effect of extracellular vesicles isolated from human liver stem cells (HLSC-EVs). Using the coincubation protocol, vesicles were loaded with the anti-tumor miRNA-145, and their effect was evaluated on renal cancer stem cell invasion. Loaded HLSC-EVs maintained their integrity and miR transfer ability. Loaded miR-145, but not miR-145 alone, was protected by RNAse digestion, possibly due to its binding to RNA-binding proteins on HLSC-EV surface, such as Annexin A2. Moreover, miR-145 coincubated HLSC-EVs were more effective in inhibiting the invasive properties of cancer stem cells, in comparison to naïve vesicles. The protocol reported here exploits a well described property of extracellular vesicles to bind nucleic acids on their surface and protect them from degradation, in order to obtain an effective miRNA loading, thus increasing the activity of therapeutically active naïve extracellular vesicles.
Whole Transcription Profile of Responders to Anti-TNF Drugs in Pediatric Inflammatory Bowel Disease
Authors: Salvador-Martín Sara , Kaczmarczyk Bartosz , Álvarez Rebeca , Navas-López Víctor Manuel , Gallego-Fernández Carmen , Moreno-Álvarez Ana , Solar-Boga Alfonso , Sánchez Cesar , Tolin Mar , Velasco Marta , Muñoz-Codoceo Rosana , Rodriguez-Martinez Alejandro , Vayo Concepción A. , Bossacoma Ferrán , Pujol-Muncunill Gemma , Fobelo María J. , Millán-Jiménez Antonio , Magallares Lorena , Martínez-Ojinaga Eva , Loverdos Inés , Eizaguirre Francisco J. , Blanca-García José A. , Clemente Susana , García-Romero Ruth , Merino-Bohórquez Vicente , González de Caldas Rafael , Vázquez Enrique , Dopazo Ana , Sanjurjo-Sáez María , López-Fernández Luis A.
Published: January 2021

Background: Up to 30% of patients with pediatric inflammatory bowel disease (IBD) do not respond to anti-Tumor Necrosis Factor (anti-TNF) therapy. The aim of this study was to identify pharmacogenomic markers that predict early response to anti-TNF drugs in pediatric patients with IBD. Methods: An observational, longitudinal, prospective cohort study was conducted. The study population comprised 38 patients with IBD aged < 18 years who started treatment with infliximab or adalimumab (29 responders and nine non-responders). Whole gene expression profiles from total RNA isolated from whole blood samples of six responders and six non-responders taken before administration of the biologic and after two weeks of therapy were analyzed using next-generation RNA sequencing. The expression of six selected genes was measured for purposes of validation in all of the 38 patients recruited using qPCR. Results: Genes were differentially expressed in non-responders and responders (32 before initiation of treatment and 44 after two weeks, Log2FC (Fold change) >0.6 or <−0.6 and p value < 0.05). After validation, FCGR1A, FCGR1B, and GBP1 were overexpressed in non-responders two weeks after initiation of anti-TNF treatment (Log2FC 1.05, 1.21, and 1.08, respectively, p value < 0.05). Conclusion: Expression of the FCGR1A, FCGR1B, and GBP1 genes is a pharmacogenomic biomarker of early response to anti-TNF agents in pediatric IBD.
Control Strategy for Process Development of High-Shear Wet Granulation and Roller Compaction to Prepare a Combination Drug Using Integrated Quality by Design
Authors: Kim Ji Yeon , Chun Myung Hee , Choi Du Hyung
Published: January 2021

In this study, we developed a control strategy for a drug product prepared by high-shear wet granulation and roller compaction using integrated quality by design (QbD). During the first and second stages, we optimized the process parameters through the design of experiments and identified the intermediate quality attributes (IQAs) and critical quality attributes (CQAs) relationship, respectively. In the first stage, we conducted an initial risk assessment by selecting critical process parameters with high impact on IQAs and CQAs and confirmed the correlation between control and response factors. Additionally, we performed Monte Carlo simulations by optimizing the process parameters to deriving and building a robust design space. In the second stage, we identified the IQAs and CQAs relationship for the control strategy, using multivariate analysis (MVA). Based on MVA, in the metformin layer, dissolution at 1 h was significantly correlated with intrinsic dissolution rate and granule size, and dissolution at 3 h was significantly correlated with bulk density and granule size. In dapagliflozin layer, dissolution at 10 min and 15 min was significantly correlated with granule size. Our results suggest that the desired drug quality may result through IQAs monitoring during the process and that the integrated QbD approach utilizing MVA can be used to develop a control strategy for producing high-quality drug products.
Strengths and Challenges of Secretory Ribonucleases as AntiTumor Agents
Authors: Castro Jessica , Ribó Marc , Vilanova Maria , Benito Antoni
Published: January 2021

Approaches to develop effective drugs to kill cancer cells are mainly focused either on the improvement of the currently used chemotherapeutics or on the development of targeted therapies aimed at the selective destruction of cancer cells by steering specific molecules and/or enhancing the immune response. The former strategy is limited by its genotoxicity and severe side effects, while the second one is not always effective due to tumor cell heterogeneity and variability of targets in cancer cells. Between these two strategies, several approaches target different types of RNA in tumor cells. RNA degradation alters gene expression at different levels inducing cell death. However, unlike DNA targeting, it is a pleotropic but a non-genotoxic process. Among the ways to destroy RNA, we find the use of ribonucleases with antitumor properties. In the last few years, there has been a significant progress in the understanding of the mechanism by which these enzymes kill cancer cells and in the development of more effective variants. All the approaches seek to maintain the requirements of the ribonucleases to be specifically cytotoxic for tumor cells. These requirements start with the competence of the enzymes to interact with the cell membrane, a process that is critical for their internalization and selectivity for tumor cells and continue with the downstream effects mainly relying on changes in the RNA molecular profile, which are not only due to the ribonucleolytic activity of these enzymes. Although the great improvements achieved in the antitumor activity by designing new ribonuclease variants, some drawbacks still need to be addressed. In the present review, we will focus on the known mechanisms used by ribonucleases to kill cancer cells and on recent strategies to solve the shortcomings that they show as antitumor agents, mainly their pharmacokinetics.
Potential Novel Food-Related and Biomedical Applications of Nanomaterials Combined with Bacteriocins
Authors: Naskar Atanu , Kim Kwang-sun
Published: January 2021

Bacteriocins are antimicrobial peptides or proteinaceous materials produced by bacteria against pathogens. These molecules have high efficiency and specificity and are equipped with many properties useful in food-related applications, such as food preservatives and additives, as well as biomedical applications, such as serving as alternatives to current antibacterial, antiviral, anticancer, and antibiofilm agents. Despite their advantages as alternative therapeutics over existing strategies, several limitations of bacteriocins, such as the high cost of isolation and purification, narrow spectrum of activity, low stability and solubility, and easy enzymatic degradation, need to be improved. Nanomaterials are promising agents in many biological applications. They are widely used in the conjugation or decoration of bacteriocins to augment the activity of bacteriocins or reduce problems related to their use in biomedical applications. Therefore, bacteriocins combined with nanomaterials have emerged as promising molecules that can be used in various biomedical applications. This review highlights the features of bacteriocins and their limitations in biomedical applications and provides a detailed overview of the uses of different nanomaterials in improving the limitations. Our review focuses on the potential applications of nanomaterials combined with bacteriocins as new designer molecules for use in future therapeutic strategies.
Iontosomes: Electroresponsive Liposomes for Topical Iontophoretic Delivery of Chemotherapeutics to the Buccal Mucosa
Authors: Sonaje Kiran , Tyagi Vasundhara , Chen Yong , Kalia Yogeshvar N.
Published: January 2021

The targeted local delivery of anticancer therapeutics offers an alternative to systemic chemotherapy for oral cancers not amenable to surgical excision. However, epithelial barrier function can pose a challenge to their passive topical delivery. The charged, deformable liposomes—“iontosomes”—described here are able to overcome the buccal mucosal barrier via a combination of the electrical potential gradient imposed by iontophoresis and their shape-deforming characteristics. Two chemotherapeutic agents with very different physicochemical properties, cisplatin (CDDP) and docetaxel (DTX), were co-encapsulated in cationic iontosomes comprising 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) and Lipoid-S75. The entrapment of CDDP was improved by formulating it in anionic reverse micelles of dipalmitoyl-sn-glycero-3-phospho-rac-glycerol sodium (DPPG) prior to loading in the iontosomes. Cryo-TEM imaging clearly demonstrated the iontosomes’ electroresponsive shape-deformable properties. The in vitro transport study using porcine mucosa indicated that iontosomes did not enter the mucosa without an external driving force. However, anodal iontophoresis resulted in significant amounts of co-encapsulated CDDP and DTX being deposited in the buccal mucosa; e.g., after current application for 10 min, the deposition of CDDP and DTX was 13.54 ± 1.78 and 10.75 ± 1.75 μg/cm2 cf. 0.20 ± 0.07 and 0.19 ± 0.09 μg/cm2 for the passive controls—i.e., 67.7- and 56.6-fold increases—without any noticeable increase in their transmucosal permeation. Confocal microscopy confirmed that the iontosomes penetrated the mucosa through the intercellular spaces and that the penetration depth could be controlled by varying the duration of current application. Overall, the results suggest that the combination of topical iontophoresis with a suitable nanocarrier system can be used to deliver multiple “physicochemically incompatible” chemotherapeutics selectively to oral cancers while decreasing the extent of systemic absorption and the associated risk of side effects.
A Drug Safety Surveillance Study of a Ciprofloxacin/Dexamethasone Ophthalmic Fixed Combination in Peruvian Population
Authors: Contreras-Salinas Homero , Baiza-Durán Leopoldo Martín , Barajas-Hernández Mariana , Vázquez-Álvarez Alan Omar , Rodríguez-Herrera Lourdes Yolotzin
Published: January 2021

(1) Background: drugs provide a significant benefit for patients who require medical treatment; however, their use implies an intrinsic potential danger, with the possibility of causing unwanted effects. These effects are known as adverse drug reactions (ADRs). Post-marketing drug safety surveillance detects unknown risks that have not been identified in clinical trials, and it is necessary to monitor marketed medications under real-life practice. Due to the scarce information about fixed combination of ciprofloxacin 0.3%/dexamethasone 0.1% (SDO), we performed a drug safety surveillance study. (2) Methods: A prospective non-controlled drug safety surveillance study was conducted in Peruvian population. A total of 236 patients prescribed SDO were included derived from 12 sites. Patients’ standardized information was collected through two phone calls, including demographics, medical history, prescribing patterns of SDO, concomitant medication, and ADRs in detail. The ADRs were classified by causality and severity, followed by outcome measures to identify new risk. (3) Results: 236 patients prescribed with SDO participated in the study and 220 were included. A total of 82 ADRs/220 patients were reported after the use of SDO, presenting a ratio 0.37 ADR/patient. The most frequent ADR with SDO administration was eye irritation (30%). All ADRs were classified as non-serious, and 97.5% (n = 80) were classified as mild while 2.5% as moderate (n = 2). No cases under the severe category were identified. (4) Conclusion: No new risks were found in the population where this study was conducted.
Simultaneous Dual-Modal Multispectral Photoacoustic and Ultrasound Macroscopy for Three-Dimensional Whole-Body Imaging of Small Animals
Authors: Park Eun-Yeong , Park Sinyoung , Lee Haeni , Kang Munsik , Kim Chulhong , Kim Jeesu
Published: January 2021

Photoacoustic imaging is a promising medical imaging technique that provides excellent function imaging of an underlying biological tissue or organ. However, it is limited in providing structural information compared to other imaging modalities, such as ultrasound imaging. Thus, to offer complete morphological details of biological tissues, photoacoustic imaging is typically integrated with ultrasound imaging. This dual-modal imaging technique is already implemented on commercial clinical ultrasound imaging platforms. However, commercial platforms suffer from limited elevation resolution compared to the lateral and axial resolution. We have successfully developed a dual-modal photoacoustic and ultrasound imaging to address these limitations, specifically targeting animal studies. The system can acquire whole-body images of mice in vivo and provide complementary structural and functional information of biological tissue information simultaneously. The color-coded depth information can be readily obtained in photoacoustic images using complementary information from ultrasound images. The system can be used for several biomedical applications, including drug delivery, biodistribution assessment, and agent testing.
Edible Chitosan/Propolis Coatings and Their Effect on Ripening, Development of Aspergillus flavus, and Sensory Quality in Fig Fruit, during Controlled Storage
Authors: Aparicio-García Pablo F. , Ventura-Aguilar Rosa I. , del Río-García Juan C. , Hernández-López Mónica , Guillén-Sánchez Dagoberto , Salazar-Piña Dolores A. , Ramos-García Margarita de L. , Bautista-Baños Silvia
Published: January 2021

Biodegradable alternatives for the control of Aspergillus flavus in fig fruit were tested with the application of coatings based on chitosan (CS) and propolis (P). To potentiate the fungicidal effect, nanoparticles of these two (CSNPs and PNPs) were also considered. The objectives of this research were to evaluate the effect of different formulations on: (a) the ripening process of the fig, (b) the incidence of A. flavus and the production of aflatoxins, and (c) the acceptance of the treated fruit by a panel. The nanostructured coatings did not influence the ripening process of the fruit during the 12 days of storage, however, the antioxidant activity increased by approximately 30% with the coating CS + PNPs + P. The figs treated with CS + CSNPs + PNPs + P, inhibited the growth of the fungus by about 20% to 30% under laboratory and semi-commercial conditions. For all treatments, the aflatoxin production was lower than 20 ppb compared to the control with values of c.a. 250 ppb. The sensory quality was acceptable among the panel. The edible coatings can be a non-toxic alternative for post-harvest preservation and the consumption of fig fruit. The next step will be its inclusion and evaluation at a commercial level in packing houses.
Wheat, Rye, and Barley Genomes Can Associate during Meiosis in Newly Synthesized Trigeneric Hybrids
Authors: Rey María-Dolores , Ramírez Carmen , Martín Azahara C.
Published: January 2021

Polyploidization, or whole genome duplication (WGD), has an important role in evolution and speciation. One of the biggest challenges faced by a new polyploid is meiosis, in particular, discriminating between multiple related chromosomes so that only homologs recombine to ensure regular chromosome segregation and fertility. Here, we report the production of two new hybrids formed by the genomes of species from three different genera: a hybrid between Aegilops tauschii (DD), Hordeum chilense (HchHch), and Secale cereale (RR) with the haploid genomic constitution HchDR (n = 7× = 21); and a hybrid between Triticum turgidum spp. durum (AABB), H. chilense, and S. cereale with the constitution ABHchR (n = 7× = 28). We used genomic in situ hybridization and immunolocalization of key meiotic proteins to establish the chromosome composition of the new hybrids and to study their meiotic behavior. Interestingly, there were multiple chromosome associations at metaphase I in both hybrids. A high level of crossover (CO) formation was observed in HchDR, which shows the possibility of meiotic recombination between the different genomes. We succeeded in the duplication of the ABHchR genome, and several amphiploids, AABBHchHchRR, were obtained and characterized. These results indicate that recombination between the genera of three economically important crops is possible.
The Use of Essential Oils from Thyme, Sage and Peppermint against Colletotrichum acutatum
Authors: Morkeliūnė Armina , Rasiukevičiūtė Neringa , Šernaitė Lina , Valiuškaitė Alma
Published: January 2021

The Colletotrichum spp. is a significant strawberry pathogen causing yield losses of up to 50%. The most common method to control plant diseases is through the use of chemical fungicides. The findings of plants antimicrobial activities, low toxicity, and biodegradability of essential oils (EO), make them suitable for biological protection against fungal pathogens. The aim is to evaluate the inhibition of Colletotrichum acutatum by thyme, sage, and peppermint EO in vitro on detached strawberry leaves and determine EO chemical composition. Our results revealed that the dominant compound of thyme was thymol 41.35%, peppermint: menthone 44.56%, sage: α,β-thujone 34.45%, and camphor: 20.46%. Thyme EO inhibited C. acutatum completely above 200 μL L−1 concentration in vitro. Peppermint and sage EO reduced mycelial growth of C. acutatum. In addition, in vitro, results are promising for biological control. The detached strawberry leaves experiments showed that disease reduction 4 days after inoculation was 15.8% at 1000 μL L−1 of peppermint EO and 5.3% at 800 μL L−1 of thyme compared with control. Our findings could potentially help to manage C. acutatum; however, the detached strawberry leaves assay showed that EO efficacy was relatively low on tested concentrations and should be increased.
An Optimized Protein Extraction Method for Gel-Free Proteomic Analysis of Opuntia Ficus-Indica
Authors: Hashiguchi Akiko , Yamaguchi Hisateru , Hitachi Keisuke , Watanabe Kazuo
Published: January 2021

Opuntia spp. is an economically important vegetable crop with high stress-tolerance and health benefits. However, proteomic analysis of the plant has been difficult due to the composition of its succulent cladodes; the abundant polysaccharides interfere with protein extraction. To facilitate proteomic analysis of this plant, we present a rapid and simple protein extraction method for Opuntia ficus-indica (L.) Miller. The optimized method produced highly reproducible protein patterns and was compatible with a gel-free quantitative workflow without the need for additional purification. We successfully analyzed the cladode mesocarp and exocarp tissues, resulting in the identification of 319 proteins. In addition, we used this method to examine the relative changes in the Opuntia proteome in response to salt stress to determine whether physiological changes could be captured. Qualified observations were obtained, revealing that salt stress increased phosphoenolpyruvate carboxylase abundance and decreased ribulose-bisphosphate carboxylase in young O. ficus-indica plants. These findings suggest that Crassulacean acid metabolism is promoted under salinity. This study highlights the efficacy of our optimized protein extraction method for elucidating the metabolic adaptations of Opuntia using gel-free proteomic analysis.
Antinociceptive Effect of an Aqueous Extract and Essential Oil from Baccharis heterophylla
Authors: Castillejos-Ramírez Erika , Pérez-Vásquez Araceli , Torres-Colín Rafael , Navarrete Andrés , Andrade-Cetto Adolfo , Mata Rachel
Published: January 2021

Infusions and poultices prepared from the aerial parts of Baccharis heterophylla Kunth (Asteraceae) are widely used in Oaxaca (Mexico) for relieving painful and inflammatory complaints. Therefore, the antinociceptive potential of an aqueous extract (31.6–316 mg/kg, p.o.) and essential oil (30–177 µg/paw, i.pl.) of the plant was assessed using the formalin test. Both preparations inhibited the formalin-induced nociception response (100–316 mg/kg and 100–177 µg/paw, respectively) during the test’s second phase. Chemical analysis of the aqueous extract revealed that the major active components were chlorogenic acid (1), 3,4-di-O-(E)-caffeoylquinic acid (2), 3,5-di-O-(E)-caffeoylquinic acid (3), 4,5-di-O-(E)-caffeoylquinic acid (4), 3,5-di-O-(E)-caffeoylquinic acid methyl ester (5), apigenin (6), genkwanin (7), acacetin (8). Compounds 1–5 and 8 are new for B. heterophylla. A high-pressure liquid chromatographic method for quantifying chlorogenic acid (1) and di-caffeoylquinic acids 2–4 in the plant was developed and validated. Analyses of the essential oil and the headspace solid-phase microextraction products, via gas-chromatography-mass spectrometry, revealed that the major volatiles were β-pinene, myrcene, D-limonene, β-caryophyllene, and α-caryophyllene, which have demonstrated antinociceptive properties.
Phytochemical Analysis and Evaluation of Antioxidant and Biological Activities of Extracts from Three Clauseneae Plants in Northern Thailand
Authors: Tanruean Keerati , Poolprasert Pisit , Suwannarach Nakarin , Kumla Jaturong , Lumyong Saisamorn
Published: January 2021

This study established the DNA barcoding sequences (matK and rbcL) of three plant species identified in the tribe Clauseneae, namely Clausena excavata, C. harmandiana and Murraya koenigii. The total phenolic and total flavonoid contents, together with the biological activities of the derived essential oils and methanol extracts, were also investigated. Herein, the success of obtaining sequences of these plant using two different barcode genes matK and rbcL were 62.5% and 100%, respectively. Both regions were discriminated by around 700 base pairs and these had resemblance with those of the Clausenae materials earlier deposited in Genbank at a 99–100% degree of identity. Additionally, the use of matK DNA sequences could positively confirm the identity as monophyletic. The highest total phenolic and total flavonoid content values (p < 0.05) were observed in the methanol extract of M. koenigii at 43.50 mg GAE/g extract and 66.13 mg QE/g extract, respectively. Furthermore, anethole was detected as the dominant compound in C. excavata (86.72%) and C. harmandiana (46.09%). Moreover, anethole (26.02%) and caryophyllene (21.15%) were identified as the major phytochemical compounds of M. koenigii. In terms of the biological properties, the M. koenigii methanol extract was found to display the greatest amount of antioxidant activity (DPPH; IC50 95.54 µg/mL, ABTS value 118.12 mg GAE/g extract, FRAP value 48.15 mg GAE/g extract), and also revealed the highest α-glucosidase and antihypertensive inhibitory activities with percent inhibition values of 84.55 and 84.95. Notably, no adverse effects on human peripheral blood mononuclear cells were observed with regard to all of the plant extracts. Furthermore, M. koenigii methanol extract exhibited promise against human lung cancer cells almost at 80% after 24 h and 90% over 48 h.
Sterols and Triterpenes from Dobera glabra Growing in Saudi Arabia and Their Cytotoxic Activity
Authors: Abdel-Mageed Wael M. , El-Gamal Ali A. , Al-Massarani Shaza M. , Basudan Omer A. , Badria Farid A. , Abdel-Kader Maged S. , Al-Rehaily Adnan J. , Aati Hanan Y.
Published: January 2021

A new lupane caffeoyl ester, lup-20(29)-ene 3β-caffeate-30-al (7), and a new oleanane-type triterpene, 3β-hydroxyolean-13(18)-en-12-one (17), were isolated from the aerial parts of Dobera glabra (Forssk), along with ten known triterpenes, including seven lupane-type lupeol (1), 30-nor-lup-3β-ol-20-one (2), ∆1-lupenone (3), lup-20(29)-en-3β,30-diol (4), lupeol caffeate (5), 30-hydroxy lup-20(29)-ene 3β-caffeate (6), and betunaldehyde (8); three oleanane-type compounds were also identified, comprising δ-amyrone (15), δ-amyrin (16), and 11-oxo-β-amyrin (18); together with six sterols, comprising β-sitosterol (9), stigmasterol (10), 7α-hydroxy-β-sitosterol (11), 7α-hydroxy-stigmasterol (12), 7-keto-β-sitosterol (13), and 7-keto-stigmasterol (14). Their structures were elucidated using a variety of spectroscopic techniques, including 1D (1H, 13C, and DEPT-135 13C) and 2D (1H–1H COSY, 1H–13C HSQC, and 1H–13C HMBC) nuclear magnetic resonance (NMR) and accurate mass spectroscopy. Subsequently, the different plant extracts and some of the isolated compounds (1–9, 11 and 13) were investigated for their possible cytotoxic activity in comparison to cisplatin against a wide array of aggressive cancer cell lines, such as colorectal cancer (HCT-116), hepatocellular carcinoma (HepG-2), and prostate cancer (PC-3) cell lines. Compound 11 displayed broad cytotoxicity against all of the tested cell lines (IC50 ≅ 8 µg/mL in all cases), and a high safety margin against normal Vero cells (IC50 = 70 µg/mL), suggesting that 11 may be a highly selective and effective anticancer agent candidate. Notably, the evidence indicated that the mode of action of compound 11 could possibly consist of the inhibition of phosphodiesterase I (80.2% enzyme inhibition observed at 2 µM compound concentration).
Genome-Wide Identification of Banana Csl Gene Family and Their Different Responses to Low Temperature between Chilling-Sensitive and Tolerant Cultivars
Authors: Yuan Weina , Liu Jing , Takáč Tomáš , Chen Houbin , Li Xiaoquan , Meng Jian , Tan Yehuan , Ning Tong , He Zhenting , Yi Ganjun , Xu Chunxiang
Published: January 2021

The cell wall plays an important role in responses to various stresses. The cellulose synthase-like gene (Csl) family has been reported to be involved in the biosynthesis of the hemicellulose backbone. However, little information is available on their involvement in plant tolerance to low-temperature (LT) stress. In this study, a total of 42 Csls were identified in Musa acuminata and clustered into six subfamilies (CslA, CslC, CslD, CslE, CslG, and CslH) according to phylogenetic relationships. The genomic features of MaCsl genes were characterized to identify gene structures, conserved motifs and the distribution among chromosomes. A phylogenetic tree was constructed to show the diversity in these genes. Different changes in hemicellulose content between chilling-tolerant and chilling-sensitive banana cultivars under LT were observed, suggesting that certain types of hemicellulose are involved in LT stress tolerance in banana. Thus, the expression patterns of MaCsl genes in both cultivars after LT treatment were investigated by RNA sequencing (RNA-Seq) technique followed by quantitative real-time PCR (qPCR) validation. The results indicated that MaCslA4/12, MaCslD4 and MaCslE2 are promising candidates determining the chilling tolerance of banana. Our results provide the first genome-wide characterization of the MaCsls in banana, and open the door for further functional studies.
GC-MS Based Identification of the Volatile Components of Six Astragalus Species from Uzbekistan and Their Biological Activity
Authors: Gad Haidy A. , Mamadalieva Nilufar Z. , Böhmdorfer Stefan , Rosenau Thomas , Zengin Gokhan , Mamadalieva Rano Z. , Al Musayeib Nawal M. , Ashour Mohamed L.
Published: January 2021

The compositions of volatile components in the aerial parts of six Astragalus species, namely A. campylotrichus (Aca), A. chiwensis (Ach), A. lehmannianus (Ale), A. macronyx (Ama), A. mucidus (Amu) and A. sieversianus (Asi), were investigated using gas chromatograph-mass spectrometry (GC-MS) analysis. Ninety-seven metabolites were identified, accounting for 73.28, 87.03, 74.38, 87.93, 85.83, and 91.39% of Aca, Ach, Ale, Ama, Amu and Asi whole oils, respectively. Sylvestrene was the most predominant component in Asi, Amu and Ama, with highest concentration in Asi (64.64%). In addition, (E)-2-hexenal was present in a high percentage in both Ale and Ach (9.97 and 10.1%, respectively). GC-MS based metabolites were subjected to principal component analysis (PCA) and hierarchal cluster analysis (HCA) to explore the correlations between the six species. The PCA score plot displayed clear differentiation of all Astragalus species and a high correlation between the Amu and Ama species. The antioxidant activity was evaluated in vitro using various assays, phosphomolybdenum (PM), 2,2 diphenyl-1-picryl-hydrazyl-hydrate (DPPH), 2,2-azino bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), cupric reducing antioxidant capacity (CUPRAC), ferric reducing power (FRAP) and ferrous ion chelation (FIC) assays. In addition, the potential for the volatile samples to inhibit both acetyl/butyrylcholinesterases (AChE, BChE), α- amylase, α-glucosidase and tyrosinase was assessed. Most of the species showed considerable antioxidant potential in the performed assays. In the DPPH assay, Ama exhibited the maximum activity (24.12 ± 2.24 mg TE/g sample), and the volatiles from Amu exhibited the highest activity (91.54 mgTE/g oil) in the ABTS radical scavenging assay. The effect was more evident in both CUPRAC and FRAP assays, where both Ale and Ama showed the strongest activity in comparison with the other tested species (84.06, 80.28 mgTE/g oil for CUPRAC and 49.47, 49.02 mgTE/g oil for FRAP, respectively). Asi demonstrated the strongest AChE (4.55 mg GALAE/g oil) and BChE (3.61 mg GALAE/g oil) inhibitory effect. Furthermore, the best tyrosinase inhibitory potential was observed for Ale (138.42 mg KAE/g). Accordingly, Astragalus species can be utilized as promising natural sources for many medicinally important components that could be tested as drug candidates for treating illnesses such as Alzheimer’s disease, diabetes mellitus and oxidative stress-related diseases.
Plants under the Attack of Allies: Moving towards the Plant Pathobiome Paradigm
Authors: Mannaa Mohamed , Seo Young-Su
Published: January 2021

Plants are functional macrobes living in a close association with diverse communities of microbes and viruses as complex systems that continuously interact with the surrounding environment. The microbiota within the plant holobiont serves various essential and beneficial roles, such as in plant growth at different stages, starting from seed germination. Meanwhile, pathogenic microbes—differentiated from the rest of the plant microbiome based on their ability to damage the plant tissues through transient blooming under specific conditions—are also a part of the plant microbiome. Recent advances in multi-omics have furthered our understanding of the structure and functions of plant-associated microbes, and a pathobiome paradigm has emerged as a set of organisms (i.e., complex eukaryotic, microbial, and viral communities) within the plant’s biotic environment which interact with the host to deteriorate its health status. Recent studies have demonstrated that the one pathogen–one disease hypothesis is insufficient to describe the disease process in many cases, particularly when complex organismic communities are involved. The present review discusses the plant holobiont and covers the steady transition of plant pathology from the one pathogen–one disease hypothesis to the pathobiome paradigm. Moreover, previous reports on model plant diseases, in which more than one pathogen or co-operative interaction amongst pathogenic microbes is implicated, are reviewed and discussed.
Red and Blue Netting Alters Leaf Morphological and Physiological Characteristics in Apple Trees
Authors: Bastías Richard M. , Losciale Pasquale , Chieco Camilla , Corelli-Grappadelli Luca
Published: January 2021

There is little information about the role of red and blue light on leaf morphology and physiology in fruit trees, and more studies have been developed in herbaceous plants grown under controlled light conditions. The objective of this research was to evaluate the effect of red and blue screens on morpho-anatomy and gas exchange in apple leaves grown under ambient sunlight conditions. Apple trees cv. Fuji were covered by 40% red and blue nets, leaving trees with 20% white net as control. Light relations (photosynthetic photon flux density, PPFD; red to far-red light ratio, R/FR and blue to red light ratio, B/R), morpho-anatomical features of the leaf (palisade to spongy mesophyll ratio, P/S, and stomata density, SD) and leaf gas exchange (net photosynthesis rate, An; stomatal conductance, gs; transpiration rate, E; and intrinsic water use efficiency, IWUE) were evaluated. Red and blue nets reduced 27% PPFD, reducing by 20% SD and 25% P/S compared to control, but without negative effects on An and gs. Blue net increased gs 21%, leading to the highest E and lowest IWUE by increment of B/R light proportion. These findings demonstrate the potential use of red and blue nets for differential modulation of apple leaf gas exchange through sunlight management under field conditions.
Piper nigrum CYP719A37 Catalyzes the Decisive Methylenedioxy Bridge Formation in Piperine Biosynthesis
Authors: Schnabel Arianne , Cotinguiba Fernando , Athmer Benedikt , Vogt Thomas
Published: January 2021

Black pepper (Piper nigrum) is among the world’s most popular spices. Its pungent principle, piperine, has already been identified 200 years ago, yet the biosynthesis of piperine in black pepper remains largely enigmatic. In this report we analyzed the characteristic methylenedioxy bridge formation of the aromatic part of piperine by a combination of RNA-sequencing, functional expression in yeast, and LC-MS based analysis of substrate and product profiles. We identified a single cytochrome P450 transcript, specifically expressed in black pepper immature fruits. The corresponding gene was functionally expressed in yeast (Saccharomyces cerevisiae) and characterized for substrate specificity with a series of putative aromatic precursors with an aromatic vanilloid structure. Methylenedioxy bridge formation was only detected when feruperic acid (5-(4-hydroxy-3-methoxyphenyl)-2,4-pentadienoic acid) was used as a substrate, and the corresponding product was identified as piperic acid. Two alternative precursors, ferulic acid and feruperine, were not accepted. Our data provide experimental evidence that formation of the piperine methylenedioxy bridge takes place in young black pepper fruits after a currently hypothetical chain elongation of ferulic acid and before the formation of the amide bond. The partially characterized enzyme was classified as CYP719A37 and is discussed in terms of specificity, storage, and phylogenetic origin of CYP719 catalyzed reactions in magnoliids and eudicots.
MicroRNA-Mediated Responses to Cadmium Stress in Arabidopsis thaliana
Authors: Pegler Joseph L. , Oultram Jackson M. J. , Nguyen Duc Quan , Grof Christopher P. L. , Eamens Andrew L.
Published: January 2021

In recent decades, the presence of cadmium (Cd) in the environment has increased significantly due to anthropogenic activities. Cd is taken up from the soil by plant roots for its subsequent translocation to shoots. However, Cd is a non-essential heavy metal and is therefore toxic to plants when it over-accumulates. MicroRNA (miRNA)-directed gene expression regulation is central to the response of a plant to Cd stress. Here, we document the miRNA-directed response of wild-type Arabidopsis thaliana (Arabidopsis) plants and the drb1, drb2 and drb4 mutant lines to Cd stress. Phenotypic and physiological analyses revealed the drb1 mutant to display the highest degree of tolerance to the imposed stress while the drb2 mutant was the most sensitive. RT-qPCR-based molecular profiling of miRNA abundance and miRNA target gene expression revealed DRB1 to be the primary double-stranded RNA binding (DRB) protein required for the production of six of the seven Cd-responsive miRNAs analyzed. However, DRB2, and not DRB1, was determined to be required for miR396 production. RT-qPCR further inferred that transcript cleavage was the RNA silencing mechanism directed by each assessed miRNA to control miRNA target gene expression. Taken together, the results presented here reveal the complexity of the miRNA-directed molecular response of Arabidopsis to Cd stress.
Detecting Mild Water Stress in Olive with Multiple Plant-Based Continuous Sensors
Authors: Marino Giulia , Scalisi Alessio , Guzmán-Delgado Paula , Caruso Tiziano , Marra Francesco Paolo , Lo Bianco Riccardo
Published: January 2021

A comprehensive characterization of water stress is needed for the development of automated irrigation protocols aiming to increase olive orchard environmental and economical sustainability. The main aim of this study is to determine whether a combination of continuous leaf turgor, fruit growth, and sap flow responses improves the detection of mild water stress in two olive cultivars characterized by different responses to water stress. The sensitivity of the tested indicators to mild stress depended on the main mechanisms that each cultivar uses to cope with water deficit. One cultivar showed pronounced day to day changes in leaf turgor and fruit relative growth rate in response to water withholding. The other cultivar reduced daily sap flows and showed a pronounced tendency to reach very low values of leaf turgor. Based on these responses, the sensitivity of the selected indicators is discussed in relation to drought response mechanisms, such as stomatal closure, osmotic adjustment, and tissue elasticity. The analysis of the daily dynamics of the monitored parameters highlights the limitation of using non-continuous measurements in drought stress studies, suggesting that the time of the day when data is collected has a great influence on the results and consequent interpretations, particularly when different genotypes are compared. Overall, the results highlight the need to tailor plant-based water management protocols on genotype-specific physiological responses to water deficit and encourage the use of combinations of plant-based continuously monitoring sensors to establish a solid base for irrigation management.
Genome-Wide Analysis and the Expression Pattern of the ERF Gene Family in Hypericum perforatum
Authors: Zhang Qian , Zhou Wen , Li Bin , Li Lin , Fu Meng , Zhou Li , Yu Xiaoding , Wang Donghao , Wang Zhezhi
Published: January 2021

Hypericum perforatum is a well-known medicinal herb currently used as a remedy for depression as it contains many high levels of secondary metabolites. The ethylene response factor (ERF) family encodes transcriptional regulators with multiple functions that play a vital role in the diverse developmental and physiological processes of plants, which can protect plants from various stresses by regulating the expression of genes. Although the function of several ERF genes from other plants has been further confirmed, H. perforatum is the first sequenced species in Malpighiales, and no information regarding the ERFs has been reported thus far. In this study, a total of 101 ERF genes were identified from H. perforatum. A systematic and thorough bioinformatic analysis of the ERF family was performed using the genomic database of H. perforatum. According to the phylogenetic tree analysis, HpERFs were further classified into 11 subfamilies. Gene ontology (GO) analysis suggested that most of the HpERFs likely participate in the biological processes of plants. The cis-elements were mainly divided into five categories, associated with the regulation of gene transcription, response to various stresses, and plant development. Further analysis of the expression patterns showed that the stress-responsive HpERFs responded to different treatments. This work systematically analyzed HpERFs using the genome sequences of H. perforatum. Our results provide a theoretical basis for further investigation of the function of stress-related ERFs in H. perforatum.
Phytotoxic Potential and Phenolic Profile of Extracts from Scrophularia striata
Authors: Mousavi Seyyed Sasan , Karami Akbar , Haghighi Tahereh Movahhed , Alizadeh Saeed , Maggi Filippo
Published: January 2021

A large number of plants produce secondary metabolites known as allelochemicals that are capable of inhibiting the germination of competitive species. This process is known as allelopathy and is mediated by several classes of chemicals, among which phenolic compounds are the most frequent. Thus, plant allelochemicals can be used to control weeds in agricultural systems. In the present work, we analyzed the phenolic profile and phytotoxic potential of different extracts (pure water or water: ethanol 50:50) from Scrophulariastriata plants that were collected from two ecological regions in Iran (Pahleh and Lizan). The total polyphenolic content (TPC), as evaluated by the Folin-Ciocolteau method, ranged from 28.3 mg/g in the aqueous extract obtained from the Lizan ecotype to 39.6 mg/g in the hydroalcoholic extract obtained from the Pahleh ecotype. Moreover, HPLC analysis was aimed at determining the content of eight phenolic compounds, namely eugenol, rosmarinic acid, hesperetin, hesperedin, trans-ferulic acid, vanillin, and caffeic acid. According to the results, rosmarinic acid appeared to be the most abundant component. The phytotoxic activities of S.striata extracts were examined on the seed germination of a crop species, Lepidium sativum, and two weeds, Chenopodium album and Malva sylvestris. All extracts showed inhibitory effects on these species. The efficiency of these inhibitory effects depended on the type of plant species, origin, and concentration of extract. The highest phytotoxic activity was caused by approximately 1% concentration of extract. The most susceptible weed was M. sylvestris. The extracts that were obtained from the Pahleh ecotype, notably the hydroalcoholic ones, showed higher phytotoxicity against L. sativum, C. album and M. sylvestris. These results encourage further studies to support the use of S. striata as a source of bioherbicides.