bluuebunny/crossref_metadata_2025 huggingface dataset:
Exploration of Neusilin® US2 as an Acceptable Filler in HPMC Matrix Systems—Comparison of Pharmacopoeial and Dynamic Biorelevant Dissolution Study
Authors: Bílik Tomáš , Vysloužil Jakub , Naiserová Martina , Muselík Jan , Pavelková Miroslava , Mašek Josef , Čopová Drahomíra , Čulen Martin , Kubová Kateřina
Published: January 2022

Modern pharmaceutical technology still seeks new excipients and investigates the further use in already known ones. An example is magnesium aluminometasilicate Neusilin® US2 (NEU), a commonly used inert filler with unique properties that are usable in various pharmaceutical fields of interest. We aimed to explore its application in hypromellose matrix systems (HPMC content 10–30%) compared to the traditionally used microcrystalline cellulose (MCC) PH 102. The properties of powder mixtures and directly compressed tablets containing individual fillers NEU or MCC, or their blend with ratios of 1.5:1, 1:1, and 0.5:1 were investigated. Besides the routine pharmaceutical testing, we have enriched the matrices’ evaluation with a biorelevant dynamic dissolution study and advanced statistical analysis. Under the USP apparatus 2 dissolution test, NEU, individually, did not provide advantages compared to MCC. The primary limitations were the burst effect increase followed by faster drug release at the 10–20% HPMC concentrations. However, the biorelevant dynamic dissolution study did not confirm these findings and showed similarities in dissolution profiles. It indicates the limitations of pharmacopoeial methods in matrix tablet development. Surprisingly, the NEU/MCC blend matrices at the same HPMC concentration showed technologically advantageous properties. Besides improved flowability, tablet hardness, and a positive impact on the in vitro drug dissolution profile toward zero-order kinetics, the USP 2 dissolution data of the samples N75M50 and N50M50 showed a similarity to those obtained from the dynamic biorelevant apparatus with multi-compartment structure. This finding demonstrates the more predictable in vivo behaviour of the developed matrix systems in human organisms.
In Silico Evaluation of Binding of 2-Deoxy-D-Glucose with Mpro of nCoV to Combat COVID-19
Authors: Raman Anirudh Pratap Singh , Kumari Kamlesh , Jain Pallavi , Vishvakarma Vijay Kumar , Kumar Ajay , Kaushik Neha , Choi Eun Ha , Kaushik Nagendra Kumar , Singh Prashant
Published: January 2022

COVID-19 has threatened the existence of humanity andthis infection occurs due to SARS-CoV-2 or novel coronavirus, was first reported in Wuhan, China. Therefore, there is a need to find a promising drug to cure the people suffering from the infection. The second wave of this viral infection was shaking the world in the first half of 2021. Drugs Controllers of India has allowed the emergency use of 2-deoxy-D-glucose (2DG) in 2021 for patients suffering from this viral infection. The potentiality of 2-deoxy-D-glucose to intervene in D-glucose metabolism exists and energy deprivation is an effective parameter to inhibit cancer cell development. Once 2DG arrives in the cells, it becomes phosphorylated to 2-deoxy-D-glucose-6-phosphate (2-DG6P), a charged molecule expressively captured inside the cells. On the other hand, 2DG lacks the ability to convert into fructose-6-phosphate, resulting in a hampering of the activity of both glucose-6-phosphate isomerase and hexokinase, and finally causing cell death. Hence, the potential and effectiveness of 2DG with the main protease (Mpro) of novel coronavirus (nCoV) should be investigated using the molecular docking and molecular dynamics (MD) simulations. The ability of 2DG to inhibit the Mpro of nCoV is compared with 2-deoxyglucose (2DAG), an acyclic molecule, and 2-deoxy-D-ribose (2DR). The binding energy of the molecules with the Mpro of nCoV is calculated using molecular docking and superimposed analysis data is obtained. The binding energy of 2DG, 2DR and 2DAG was −2.40, −2.22 and −2.88 kcal/mol respectively. Although the molecular docking does not provide reliable information, therefore, the binding affinity can be confirmed by molecular dynamics simulations. Various trajectories such as Rg, RMSD, RMSF, and hydrogen bonds are obtained from the molecular dynamics (MD) simulations. 2DG was found to be a better inhibitor than the 2DAG and 2DR based on the results obtained from the MD simulations at 300 K. Furthermore, temperature-dependent MD simulations of the Mpro of nCoV with promising 2DG was performed at 295, 310 and 315 K, and the effective binding with the Mpro of nCoV occurred at 295 K. With the use of DFT calculations, optimized geometry and localization of electron density of the frontier molecular orbitals were calculated.
Gene Therapy Using Nanocarriers for Pancreatic Ductal Adenocarcinoma: Applications and Challenges in Cancer Therapeutics
Authors: Won Eun-Jeong , Park Hyeji , Yoon Tae-Jong , Cho Young-Seok
Published: January 2022

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers worldwide, and its incidence is increasing. PDAC often shows resistance to several therapeutic modalities and a higher recurrence rate after surgical treatment in the early localized stage. Combination chemotherapy in advanced pancreatic cancer has minimal impact on overall survival. RNA interference (RNAi) is a promising tool for regulating target genes to achieve sequence-specific gene silencing. Here, we summarize RNAi-based therapeutics using nanomedicine-based delivery systems that are currently being tested in clinical trials and are being developed for the treatment of PDAC. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) genome editing has been widely used for the development of cancer models as a genetic screening tool for the identification and validation of therapeutic targets, as well as for potential cancer therapeutics. This review discusses current advances in CRISPR/Cas9 technology and its application to PDAC research. Continued progress in understanding the PDAC tumor microenvironment and nanomedicine-based gene therapy will improve the clinical outcomes of patients with PDAC.
Impact of circ-0000221 in the Pathogenesis of Hepatocellular via Modulation of miR-661–PTPN11 mRNA Axis
Authors: Matboli Marwa , Hassan Mohmed Kamal , Ali Mahmoud A. , Mansour Mohamed Tarek , Elsayed Waheba , Atteya Reham , Aly Hebatallah Said , Meteini Mahmoud El , Elghazaly Hesham , El-Khamisy Sherif , Agwa Sara H. A.
Published: January 2022

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related death in Egypt. A deep understanding of the molecular events occurring in HCC can facilitate the development of novel diagnostic and/or therapeutic approaches. In the present study, we describe a novel axis of hsa-circ-0000221–miR-661–PTPN11 mRNA proposed by in silico and in vitro analysis and its role in HCC pathogenesis. We observe a reduction in the expression levels of hsa-circ-0000221 and PTPN11 mRNA in HCC patients’ sera tested compared with control subjects. The reduction occurs with a concomitant increase in the expression of miR-661. Furthermore, the introduction of exogenous hsa-circ-0000221 into Hep-G2 or SNU449 cell lines results in detectable decrease in cellular viability and an increase in apoptotic manifestations that is associated with G1 accumulation and CCDN1 overexpression. Altogether, these findings indicate the tumor-suppressive role of hsa-circ-0000221 in HCC, which acts through miR-661 inhibition, along with a subsequent PTPN11 mRNA increase, where PTPN11 is known to inhibit cell proliferation in many forms of cancer. Our study encourages further investigation of the role of circRNAs in cancer and their potential use as molecular biomarkers.
Eradication of Myrosinase-Tethered Cancer Cells by Allyl Isothiocyanate Derived from Enzymatic Hydrolysis of Sinigrin
Authors: Tarar Ammar , Alyami Esmael M. , Peng Ching-An
Published: January 2022

Sinigrin is present in significant amounts in cruciferous vegetables. Epidemiological studies suggest that the consumption of such vegetables decreases the risk of cancer, and the effect is attributed mainly to allyl isothiocyanate (AITC), a hydrolysis product of sinigrin catalyzed by myrosinase. Anticancer activity of AITC has been previously investigated for several cancer models, but less attention was paid to delivering AITC on the target site. In this study, the gene sequences of core streptavidin (coreSA) and myrosinase (MYR) were cloned in a pET-30a(+) plasmid and transformed into BL21(DE3) E. coli competent cells. The MYR-coreSA chimeric protein was expressed and purified using immobilized metal affinity chromatography and further characterized by gel electrophoresis, Western blot, and enzyme activity assay. The purified MYR-coreSA chimeric protein was tethered on the outer membrane of biotinylated adenocarcinoma A549 cells and then treated with various concentrations of sinigrin. Our results showed that 20 µM of sinigrin inhibited the growth of A549 cells tethered with myrosinase by ~60% in 48 h. Furthermore, the levels of treated cells undertaken apoptosis were determined by Caspase-3/7 activation and Annexin-V. In summary, sinigrin harnessed like a prodrug catalyzed by myrosinase to the production of AITC, which induced cell apoptosis and arrested the growth of lung cancer cells.
Polysaccharide 3D Printing for Drug Delivery Applications
Authors: Zamboulis Alexandra , Michailidou Georgia , Koumentakou Ioanna , Bikiaris Dimitrios N.
Published: January 2022

3D printing, or additive manufacturing, has gained considerable interest due to its versatility regarding design as well as in the large choice of materials. It is a powerful tool in the field of personalized pharmaceutical treatment, particularly crucial for pediatric and geriatric patients. Polysaccharides are abundant and inexpensive natural polymers, that are already widely used in the food industry and as excipients in pharmaceutical and cosmetic formulations. Due to their intrinsic properties, such as biocompatibility, biodegradability, non-immunogenicity, etc., polysaccharides are largely investigated as matrices for drug delivery. Although an increasing number of interesting reviews on additive manufacturing and drug delivery are being published, there is a gap concerning the printing of polysaccharides. In this article, we will review recent advances in the 3D printing of polysaccharides focused on drug delivery applications. Among the large family of polysaccharides, the present review will particularly focus on cellulose and cellulose derivatives, chitosan and sodium alginate, printed by fused deposition modeling and extrusion-based printing.
Targeted EV to Deliver Chemotherapy to Treat Triple-Negative Breast Cancers
Authors: Si Yingnan , Chen Kai , Ngo Hanh Giai , Guan Jia Shiung , Totoro Angela , Zhou Zhuoxin , Kim Seulhee , Kim Taehyun , Zhou Lufang , Liu Xiaoguang
Published: January 2022

Triple-negative breast cancers (TNBCs) are heterogeneous and metastatic, and targeted therapy is highly needed for TNBC treatment. Recent studies showed that extracellular vesicles (EV) have great potential to deliver therapies to treat cancers. This study aimed to develop and evaluate a natural compound, verrucarin A (Ver-A), delivered by targeted EV, to treat TNBC. First, the surface expression of epidermal growth factor receptor (EGFR) and CD47 were confirmed with immunohistochemistry (IHC) staining of patient tissue microarray, flow cytometry and Western blotting. EVs were isolated from HEK 293F culture and surface tagged with anti-EGFR/CD47 mAbs to construct mAb-EV. The flow cytometry, confocal imaging and live-animal In Vivo Imaging System (IVIS) demonstrated that mAb-EV could effectively target TNBC and deliver the drug. The drug Ver-A, with dosage-dependent high cytotoxicity to TNBC cells, was packed in mAb-EV. The anti-TNBC efficacy study showed that Ver-A blocked tumor growth in both 4T1 xenografted immunocompetent mouse models and TNBC patient-derived xenograft models with minimal side effects. This study demonstrated that the targeted mAb-EV-Ver-A had great potential to treat TNBCs.
Granzyme B PET Imaging in Response to In Situ Vaccine Therapy Combined with αPD1 in a Murine Colon Cancer Model
Authors: Hartimath Siddesh V. , Ramasamy Boominathan , Xuan Tan Yun , Rong Tang Jun , Khanapur Shivashankar , Cheng Peter , Hwang You Yi , Robins Edward G. , Goggi Julian L.
Published: January 2022

Immune checkpoint inhibitors (ICIs) block checkpoint receptors that tumours use for immune evasion, allowing immune cells to target and destroy cancer cells. Despite rapid advancements in immunotherapy, durable response rates to ICIs remains low. To address this, combination clinical trials are underway assessing whether adjuvants can enhance responsiveness by increasing tumour immunogenicity. CpG-oligodeoxynucleotides (CpG-ODN) are synthetic DNA fragments containing an unmethylated cysteine-guanosine motif that stimulate the innate and adaptive immune systems by engaging Toll-like receptor 9 (TLR9) present on the plasmacytoid dendritic cells (pDCs) and B cells. Here, we have assessed the ability of AlF-mNOTA-GZP, a peptide tracer targeting granzyme B, to serve as a PET imaging biomarker in response to CpG-ODN 1585 in situ vaccine therapy delivered intratumourally (IT) or intraperitoneally (IP) either as monotherapy or in combination with αPD1. [18F]AlF-mNOTA-GZP was able to differentiate treatment responders from non-responders based on tumour uptake. Furthermore, [18F]AlF-mNOTA-GZP showed positive associations with changes in tumour-associated lymphocytes expressing GZB, namely GZB+ CD8+ T cells, and decreases in suppressive F4/80+ cells. [18F]AlF-mNOTA-GZP tumour uptake was mediated by GZB expressing CD8+ cells and successfully stratifies therapy responders from non-responders, potentially acting as a non-invasive biomarker for ICIs and combination therapy evaluation in a clinical setting.
Revisiting Excitotoxicity in Traumatic Brain Injury: From Bench to Bedside
Authors: Baracaldo-Santamaría Daniela , Ariza-Salamanca Daniel Felipe , Corrales-Hernández María Gabriela , Pachón-Londoño Maria José , Hernandez-Duarte Isabella , Calderon-Ospina Carlos-Alberto
Published: January 2022

Traumatic brain injury (TBI) is one of the leading causes of morbidity and mortality. Consequences vary from mild cognitive impairment to death and, no matter the severity of subsequent sequelae, it represents a high burden for affected patients and for the health care system. Brain trauma can cause neuronal death through mechanical forces that disrupt cell architecture, and other secondary consequences through mechanisms such as inflammation, oxidative stress, programmed cell death, and, most importantly, excitotoxicity. This review aims to provide a comprehensive understanding of the many classical and novel pathways implicated in tissue damage following TBI. We summarize the preclinical evidence of potential therapeutic interventions and describe the available clinical evaluation of novel drug targets such as vitamin B12 and ifenprodil, among others.
Dendrimers and Dendritic Materials against Infectious Diseases
Authors: de La Mata Francisco Javier , Ortega Paula , García-Gallego Sandra
Published: January 2022

The COVID-19 pandemic showed more deeply the need of our society to provide new therapeutic strategies to fight infectious diseases, not only against currently known illnesses, where common antibiotics and drugs appear to be not fully effective, but also against new infectious threats that may arise [...]
Optimization of Photosynthetic Photon Flux Density and Light Quality for Increasing Radiation-Use Efficiency in Dwarf Tomato under LED Light at the Vegetative Growth Stage
Authors: Ke Xinglin , Yoshida Hideo , Hikosaka Shoko , Goto Eiji
Published: January 2022

Dwarf tomatoes are advantageous when cultivated in a plant factory with artificial light because they can grow well in a small volume. However, few studies have been reported on cultivation in a controlled environment for improving productivity. We performed two experiments to investigate the effects of photosynthetic photon flux density (PPFD; 300, 500, and 700 μmol m−2 s−1) with white light and light quality (white, R3B1 (red:blue = 3:1), and R9B1) with a PPFD of 300 μmol m−2 s−1 on plant growth and radiation-use efficiency (RUE) of a dwarf tomato cultivar (‘Micro-Tom’) at the vegetative growth stage. The results clearly demonstrated that higher PPFD leads to higher dry mass and lower specific leaf area, but it does not affect the stem length. Furthermore, high PPFD increased the photosynthetic rate (Pn) of individual leaves but decreased RUE. A higher blue light proportion inhibited dry mass production with the same intercepted light because the leaves under high blue light proportion had low Pn and photosynthetic light-use efficiency. In conclusion, 300 μmol m−2 s−1 PPFD and R9B1 are the recommended proper PPFD and light quality, respectively, for ‘Micro-Tom’ cultivation at the vegetative growth stage to increase the RUE.
Mineral Fertilization Influences the Growth, Cryptolepine Yield, and Bioefficacy of Cryptolepis sanguinolenta (Lindl.) Schlt.
Authors: Amissah Jacqueline Naalamle , Alorvor Forgive Enyonam , Okorley Benjamin Azu , Asare Chris Mpere , Osei-Safo Dorcas , Appiah-Opong Regina , Addae-Mensah Ivan
Published: January 2022

Cryptolepis sanguinolenta (Lindl.) Schlt., the main source of cryptolepine alkaloid, is intensively exploited in the wild to treat malaria and Lyme disease. In this study, the influence of four inorganic fertilizers (supplying N, P, K, or NPK) and four growth periods (3, 6, 9, and 12 months after transplanting) on the herb’s root biomass, cryptolepine content and yield, and biological activities were investigated in a pot and field trial. The results showed the application of N (in the form of Urea or NPK) increased root biomass yield, cryptolepine content, and cryptolepine yield compared to unfertilized plants. The 9-month-old plants recorded the maximum cryptolepine content (2.26 mg/100 mg dry root) and cryptolepine yield (304.08 mg/plant), indicating the perfect time to harvest the herb. Plant age at harvest had a more significant influence (50.6–55.7%) on cryptolepine production than fertilizer application (29.2–33.3%). Cryptolepine extracts from 9- to 12-month-old plants had the highest antiplasmodial activity (IC50 = 2.56–4.65 µg/mL) and drug selectivity index (2.15–3.91) against Plasmodium falciparum Dd2. These extracts were also cytotoxic to Jurkat leukaemia cell lines (CC50 < 62.56 µg/mL), indicating the possible use of cryptolepine for cancer management. Growing the herb in the field increased cryptolepine yield 2.5 times compared to growth in a pot, but this did not influence the antiplasmodial activity of the extract. Commercial cultivation of C. sanguinolenta for 9 months combined with N application could be a promising solution to the sustainable use of this threatened medicinal species.
Optimization of Biomass Accumulation and Production of Phenolic Compounds in Callus Cultures of Rhodiola rosea L. Using Design of Experiments
Authors: Erst Anna A. , Petruk Anastasia A. , Erst Andrey S. , Krivenko Denis A. , Filinova Nadezhda V. , Maltseva Svetlana Y. , Kulikovskiy Maxim S. , Banaev Evgeny V.
Published: January 2022

Rhodiola rosea L. is a valuable medicinal plant with adaptogenic, neuroprotective, antitumor, cardioprotective, and antidepressant effects. In this study, design of experiments methodology was employed to analyze and optimize the interacting effects of mineral compounds (concentration of NO3− and the ratio of NH4+ to K+) and two plant growth regulators [total 6-benzylaminopurine (BAP) and α-naphthylacetic acid (NAA) concentration and the ratio of BAP to NAA] on the growth and the production of total phenolic compounds (TPCs) in R. rosea calluses. The overall effect of the model was highly significant (p < 0.0001), indicating that NH4+, K+, NO3−, BAP, and NAA significantly affected growth. The best callus growth (703%) and the highest production of TPCs (75.17 mg/g) were achieved at an NH4+/K+ ratio of 0.33 and BAP/NAA of 0.33, provided that the concentration of plant growth regulators was 30 μM and that of NO3− was ≤40 mM. According to high-performance liquid chromatography analyses of aerial parts (leaves and stems), in vitro seedlings and callus cultures of R. rosea contain no detectable rosarin, rosavin, rosin, and cinnamyl alcohol. This is the first report on the creation of an experiment for the significant improvement of biomass accumulation and TPC production in callus cultures of R. rosea.
Genetic and Morpho-Agronomic Characterization of Sicilian Tetraploid Wheat Germplasm
Authors: Sicilia Angelo , Anastasi Umberto , Bizzini Michele , Montemagno Stefania , Nicotra Carmelo , Blangiforti Sebastiano , Spina Alfio , Cosentino Salvatore Luciano , Lo Piero Angela Roberta
Published: January 2022

Cereal landraces are a very valuable resource in contemporary agriculture. A renewed focus for breeding purposes could ameliorate some negative consequences of modern agriculture and conventional breeding, such as the loss of genetic diversity. One strategy combining molecular genotyping and characterization of morpho-agronomic traits related to productivity is proposed to assess a group of tetraploid wheat landraces named Bufala, historically cultivated in the mountain areas of Sicily and characterized by adaptability in terms of cold tolerance, ability to grow in marginal soils, weed competitiveness and resistance to diseases. A total of 55 SSR molecular markers were used to detect patterns of diversity in 30 rivet and durum wheat genotypes. Furthermore, phenotyping was then conducted for 8 morpho-agronomic traits. Discriminant analysis of principal components (DAPC), STRUCTURE and phylogenetical analysis allowed to identify three groups, two of them genetically close and including both Bufala and Bufala-related rivet landraces. To the third group, old and more recent durum wheat varieties, constituting the outgroup, were assigned. Clustering was confirmed by Principal Component Analysis (PCA). Finally, a correlation analysis showed that Bufala genotypes are characterized by lower ear density, major ear length and later earing time compared with the other studied genotypes. The levels of diversity and population structure could be an important contribution to parent selection in tetraploid wheat breeding programs, as well as to germplasm conservation and management.
Polyploidization Increases the Lipid Content and Improves the Nutritional Quality of Rice
Authors: Wang Wei , Tu Qiang , Chen Rongrong , Lv Pincang , Xu Yanqing , Xie Qian , Song Zhaojian , He Yuchi , Cai Detian , Zhang Xianhua
Published: January 2022

Plant polyploidization is frequently associated with changes in nutrient contents. However, the possible contribution of metabolites to this change has not been investigated by characterizing the metabolite contents of diploid and tetraploid forms of rice (Oryza sativa L.). We compared the metabolites of a group of diploid–tetraploid japonica brown rice and a group of diploid–tetraploid indica brown rice based on liquid chromatography–tandem mass spectrometry. In total, 401 metabolites were identified; of these, between the two diploid–tetraploid groups, 180 showed opposite expression trends, but 221 showed the same trends (147 higher abundance vs. 74 lower abundance). Hierarchical cluster analysis of differential metabolites between diploid and tetraploid species showed a clear grouping pattern, in which the expression abundance of lipids, amino acids and derivatives, and phenolic acids increased in tetraploids. Further analysis revealed that the lipids in tetraploid rice increased significantly, especially unsaturated fatty acids and phospholipids. This study provides further basis for understanding the changes in rice nutritional quality following polyploidization and may serve as a new theoretical reference for breeding eutrophic or functional rice varieties via polyploidization.
Use of Biostimulants: Towards Sustainable Approach to Enhance Durum Wheat Performances
Authors: Ayed Sourour , Bouhaouel Imen , Jebari Hayet , Hamada Walid
Published: January 2022

The use of biostimulant (BS) holds a promising and environmental-friendly innovation to address current needs of sustainable agriculture. The aim of the present study is twofold: (i) assess the potential of durum wheat seed coating with microbial BS (‘Panoramix’, Koppert), a mix of Bacillus spp., Trichoderma spp., and endomycorrhiza, compared to two chemical products (‘Spectro’ and ‘Mycoseeds’) through germination bioassay, pot and field trials under semi-arid conditions, and (ii) identify the most effective method of BS supply (‘seed coating’, ‘foliar spray’, and ‘seed coating + foliar spray’) under field conditions. For this purpose, three modern durum wheat cultivars were tested. ‘Panoramix’ was the most efficient treatment and enhanced all germination (germination rate, and coleoptile and radicle length), physiological (relative water content, chlorophyll content, and leaf area), and agro-morphological (plant height, biomass, seed number per spike, thousand kernel weight, and grain yield) attributes. Unexpectedly, the individual application of ‘Panoramix’ showed better performance than the combined treatment ‘Panoramix + Spectro’. Considering the physiological and agro-morphological traits, the combined method ‘seed coating + foliar spray’ displayed the best results. Principal component analysis confirmed the superiority of ‘Panoramix’ treatment or ‘seed coating + foliar spray’ method. Among tested durum wheat cultivars, ‘Salim’ performed better especially under ‘Panoramix’ treatment, but in some case ‘Karim’ valorized better this BS showing the highest increase rates. Based on these study outcomes, ‘Panoramix’ might be used as promising sustainable approach to stimulate durum wheat performance.
LC-MS Characterization and Biological Activities of Cuban Cultivars of Plectranthus neochilus Schltr
Authors: Rodríguez-Ferreiro Annarli O. , Ochoa-Pacheco Ania , Méndez-Rodriguez Daniel , Ortiz-Beatón Emilia , Font-Salmo Oneida , Guisado-Bourzac Frenkel , Molina-Bertrán Silvia , Monzote Lianet , Cos Paul , Foubert Kenn , Pieters Luc , Perez-Novo Claudina , Vanden Berghe Wim , Escalona-Arranz Julio C. , Setzer William N.
Published: January 2022

Plectranthus neochilus Schltr. (Lamiaceae) is a plant recently introduced in Cuba. Worldwide, it is an ethnomedicinal alternative for its use against microbial infections, but the Cuban population use the extracts to treat sleep disorders. To address this apparent incongruity, four collections (from different seasonal conditions in the year) of Cuban P. neochilus cultivars were analyzed in terms of their pharmacognostic characteristics. Three extracts using fresh and dried leaves were chemically and biologically characterized. UPLC-DAD-MS/MS analysis was performed to determine their chemical composition, while a panel of nine microorganisms was used to evaluate their antimicrobial activity. Finally, cytotoxic effects of different fractions were measured in three cell lines by the resazurin viability assay. In contrast to previously reported micro and macromorphological properties of P. neochilus, the leaves from the Cuban cultivars did not present glandular trichomes, nor did they produce quantifiable levels of essential oils. Moreover, aqueous extracts used by the population revealed no significant antimicrobial activity and were not cytotoxic. The three extracts showed a similar phytochemical composition, i.e., eight flavonoids, seven abietane diterpenes, and rosmarinic acid as the major constituent, most of them reported for the first time in this species. The low yield of essential oil, the absence of glandular trichomes, compounds with a high level of oxidation, and a moderate antimicrobial activity detected were the most distinctive pharmacognostic and biological characteristics of P. neochilus grown in Cuba. These aspects could explain its non-use as an antimicrobial.
Comparative Study on Phenolic Compounds and Antioxidant Activities of Hop (Humulus lupulus L.) Strobile Extracts
Authors: Lyu Jae Il , Ryu Jaihyunk , Seo Kyoung-Sun , Kang Kyung-Yun , Park Sang Hoon , Ha Tae Hyun , Ahn Joon-Woo , Kang Si-Yong
Published: January 2022

In this study, we investigated the phenolic compounds in hop strobile extracts and evaluated their antioxidant property using DPPH and ABTS assay. The total phenolic compound (TPC) and total flavonoid compound (TFC) estimated in two different solvent extracts considerably varied depending on the extraction solvent. The most abundant phenolic compound in hop strobile was humulones (α-acid) with levels ranging from 50.44 to 193.25 µg/g. El Dorado accession revealed higher antioxidant activity in ethanol extracts (DPPH: IC50 124.3 µg/mL; ABTS: IC50 95.4 µg/mL) when compared with that of the other accessions. Correlations between DPPH (IC50) scavenging TFC in ethanol extract (TFC_E, −0.941), and TPC_E (−0.901), and between ABTS (IC50) scavenging TFC_E (−0.853), and TPC_E (−0.826), were statistically significant at p < 0.01 level, whereas no significant correlation was observed between antioxidant activities, TPC and TFC in water extract. This study is the first to report that variations in the level of phenolic contents and antioxidant activity of various hop cultivars depended on the type of extraction solvent used and the cultivation regions. These results could provide valuable information on developing hop products.
Challenges in Medicinal and Aromatic Plants DNA Barcoding—Lessons from the Lamiaceae
Authors: Nazar Nazia , Howard Caroline , Slater Adrian , Sgamma Tiziana
Published: January 2022

The potential value of DNA barcoding for the identification of medicinal plants and authentication of traded plant materials has been widely recognized; however, a number of challenges remain before DNA methods are fully accepted as an essential quality control method by industry and regulatory authorities. The successes and limitations of conventional DNA barcoding are considered in relation to important members of the Lamiaceae. The mint family (Lamiaceae) contains over one thousand species recorded as having a medicinal use, with many more exploited in food and cosmetics for their aromatic properties. The family is characterized by a diversity of secondary products, most notably the essential oils (EOs) produced in external glandular structures on the aerial parts of the plant that typify well-known plants of the basil (Ocimum), lavender (Lavandula), mint (Mentha), thyme (Thymus), sage (Salvia) and related genera. This complex, species-rich family includes widely cultivated commercial hybrids and endangered wild-harvested traditional medicines, and examples of potential toxic adulterants within the family are explored in detail. The opportunities provided by next generation sequencing technologies to whole plastome barcoding and nuclear genome sequencing are also discussed with relevant examples.
An R2R3-MYB Transcription Factor OsMYBAS1 Promotes Seed Germination under Different Sowing Depths in Transgenic Rice
Authors: Wang Xiaomin , Wu Rong , Shen Tongshu , Li Zhenan , Li Chengyong , Wu Bangkui , Jiang Hongye , Zhao Guangwu
Published: January 2022

MYB-type transcription factors play essential regulatory roles in seed germination and the response to seedling establishment stress. This study isolated a rice R2R3-MYB gene, OsMYBAS1, and functionally characterized its role in seed germination by generating transgenic rice plants with the overexpression and knockout of OsMYBAS1. Gene expression analysis suggested that OsMYBAS1 was highly expressed in brown rice and root, respectively. Subcellular localization analysis determined that OsMYBAS1 was localized in the nucleus. No significant differences in seed germination rate were observed among wild-type (WT) and transgenic rice plants at the 0-cm sowing depth. However, when sown at a depth of 4 cm, higher germination rates, root lengths and seedling heights were obtained in OsMYBAS1-overexpressing plants than in WT. Furthermore, the opposite results were recorded between the osmybas1 mutants and WT. Moreover, OsMYBAS1-overexpressing plants significantly enhanced superoxide dismutase (SOD) enzyme activity and suppressed the accumulation of malondialdehyde (MDA) content at the 4-cm sowing depth. These results indicate that the MYB transcription factor OsMYBAS1 may promote rice seed germination and subsequent seedling establishment under deep-sowing conditions. These findings can provide valuable insight into rice seed-quality breeding to facilitate the development of a dry, direct-seeding production system.
Impact of Harvest Time and Pruning Technique on Total CBD Concentration and Yield of Medicinal Cannabis
Authors: Crispim Massuela Danilo , Hartung Jens , Munz Sebastian , Erpenbach Federico , Graeff-Hönninger Simone
Published: January 2022

The definition of optimum harvest and pruning interventions are important factors varying inflorescence yield and cannabinoid composition. This study investigated the impact of (i) harvest time (HT) and (ii) pruning techniques (PT) on plant biomass accumulation, CBD and CBDA-concentrations and total CBD yield of a chemotype III medical cannabis genotype under indoor cultivation. The experiment consisted of four HTs between 5 and 11 weeks of flowering and three PTs-apical cut (T); removal of side shoots (L) and control (C), not pruned plants. Results showed that inflorescence dry weight increased continuously, while the total CBD concentration did not differ significantly over time. For the studied genotype, optimum harvest time defined by highest total CBD yield was found at 9 weeks of flowering. Total CBD-concentration of inflorescences in different fractions of the plant’s height was significantly higher in the top (9.9%) in comparison with mid (8.2%) and low (7.7%) fractions. The T plants produced significantly higher dry weight of inflorescences and leaves than L and C. Total CBD yield of inflorescences for PTs were significantly different among pruned groups, but do not differ from the control group. However, a trend for higher yields was observed (T > C > L).
A Descriptive Chemical Composition of Concentrated Bud Macerates through an Optimized SPE-HPLC-UV-MS2 Method—Application to Alnus glutinosa, Ribes nigrum, Rosa canina, Rosmarinus officinalis and Tilia tomentosa
Authors: Charpentier Thomas , Boisard Séverine , Le Ray Anne-Marie , Bréard Dimitri , Chabrier Amélie , Esselin Hélène , Guilet David , Ripoll Christophe , Richomme Pascal
Published: January 2022

Concentrated bud macerates (CBMs) are obtained from meristematic tissues such as buds and young shoots by maceration in a solvent composed of glycerin, water and ethanol (1/1/1/, v/v). Their traditional utilization in gemmotherapy has gained interest in the past years, and the knowledge of their chemical characterization can provide commercial arguments, particularly to secure their quality control. Therefore, an optimized method for phytochemical analysis including glycerol removal by a preliminary solid phase extraction (SPE) followed by compound identification using high performance liquid chromatography coupled with ultra-violet and tandem mass detectors (HPLC-UV-MS2) was developed. This method was applied on 5 CBMs obtained from Alnus glutinosa, Ribesnigrum, Rosmarinus officinalis, Rosa canina and Tilia tomentosa in order to determinate their chemical composition. Their antioxidant effects were also investigated by radical scavenging activity assays (DPPH and ORAC). Glycerol removal improved the resolution of HPLC chemical profiles and allowed us to perform TLC antioxidant screening. Our approach permitted the identification of 57 compounds distributed in eight major classes, three of them being common to all macerates including nucleosides, phenolic acids and glycosylated flavonoids. Quantification of the later class as a rutin equivalent (RE) showed a great disparity between Rosa canina macerate (809 mg RE/L), and the other ones (from 175 to 470 mg RE/L). DPPH and ORAC assays confirmed the great activity of Rosa canina (4857 and 6479 μmol TE/g of dry matter, respectively). Finally, phytochemical and antioxidant analysis of CBMs strengthened their phytomedicinal interest in the gemmotherapy field.
Genome-Wide Analysis of Multiple Organellar RNA Editing Factor (MORF) Family in Kiwifruit (Actinidia chinensis) Reveals Its Roles in Chloroplast RNA Editing and Pathogens Stress
Authors: Xiong Yuhong , Fang Jing , Jiang Xiaohan , Wang Tengfei , Liu Kangchen , Peng Huixiang , Zhang Xiujun , Zhang Aidi
Published: January 2022

Kiwifruit (Actinidia chinensis) is well known for its high vitamin C content and good taste. Various diseases, especially bacterial canker, are a serious threat to the yield of kiwifruit. Multiple organellar RNA editing factor (MORF) genes are pivotal factors in the RNA editosome that mediates Cytosine-to-Uracil RNA editing, and they are also indispensable for the regulation of chloroplast development, plant growth, and response to stresses. Although the kiwifruit genome has been released, little is known about MORF genes in kiwifruit at the genome-wide level, especially those involved in the response to pathogens stress. In this study, we identified ten MORF genes in the kiwifruit genome. The genomic structures and chromosomal locations analysis indicated that all the MORF genes consisted of three conserved motifs, and they were distributed widely across the seven linkage groups and one contig of the kiwifruit genome. Based on the structural features of MORF proteins and the topology of the phylogenetic tree, the kiwifruit MORF gene family members were classified into six groups (Groups A–F). A synteny analysis indicated that two pairs of MORF genes were tandemly duplicated and five pairs of MORF genes were segmentally duplicated. Moreover, based on analysis of RNA-seq data from five tissues of kiwifruit, we found that both expressions of MORF genes and chloroplast RNA editing exhibited tissue-specific patterns. MORF2 and MORF9 were highly expressed in leaf and shoot, and may be responsible for chloroplast RNA editing, especially the ndhB genes. We also observed different MORF expression and chloroplast RNA editing profiles between resistant and susceptible kiwifruits after pathogen infection, indicating the roles of MORF genes in stress response by modulating the editing extend of mRNA. These results provide a solid foundation for further analyses of the functions and molecular evolution of MORF genes, in particular, for clarifying the resistance mechanisms in kiwifruits and breeding new cultivars with high resistance.
Berries and Leaves of Actinidia kolomikta (Rupr. & Maxim.) Maxim.: A Source of Phenolic Compounds
Authors: Česonienė Laima , Štreimikytė Paulina , Liaudanskas Mindaugas , Žvikas Vaidotas , Viškelis Pranas , Viškelis Jonas , Daubaras Remigijus
Published: January 2022

Berries of Actinidia kolomikta (A. kolomikta) are known for high ascorbic acid content, but the diversity of phenolic compounds has been little studied. The present research aimed to investigate phenolic compounds and antioxidant activity in berries and leaves of twelve A. kolomikta cultivars. The UHPLC-ESI-MS/MS technique was used to determine differences among cultivars in the quantitative composition of individual phenolic compounds. Antioxidant activity was determined by DPPH• free radical scavenging and CUPRAC methods. In the present study, 13 phenolic compounds were detected in berries, whereas leaves contained 17 phenolic compounds. Flavonols were the primary class found in both berries and leaves; other identified phenolic compounds were flavan-3-ols, flavones and, phenolic acids; and dihydrochalcone phloridzin was identified in the leaves. The amount and variety of phenolic compounds in berries and leaves and antioxidant activity were found to be cultivar-dependent. The highest total content of phenolic compounds was found in the leaves of the cultivar ‘Aromatnaja’ and in the berries of the cultivar ‘VIR-2’. Results of this study have confirmed that berries and leaves of A. kolomikta could be a valuable raw material for both food and pharmaceutical industries.
Organic Amendments Effects on Nutrient Uptake, Secondary Metabolites, and Antioxidant Properties of Melastoma malabathricum L.
Authors: Rusli Lili Syahani , Abdullah Rosazlin , Yaacob Jamilah Syafawati , Osman Normaniza
Published: January 2022

Amelioration of soil acidity can boost soil fertility, hence increasing nutrient uptake, secondary metabolite, and its antioxidant potential. In the present study, the effectiveness of food waste compost and palm kernel biochar was assessed as soil amendments for Melastoma malabathricum L. grown in acidic soil conditions. A six-month greenhouse study was conducted using completely randomized design (CRD) with three treatment groups, including control plants (T1), plants amended with palm kernel biochar (T2), and plants amended with food waste compost (T3). Data analysis revealed that Melastoma malabathricum L. amended with T3 recorded the highest total chlorophyll content (433.678 ± 13.224 µg g−1 DW), followed by T2 and T1. The increase in chlorophyll content was contributed by the increase in soil pH. This was shown by the positive significant correlations between soil pH and chlorophyll a (r2 = 0.96; p ≤ 0.01) and chlorophyll b (r2 = 0.778; p ≤ 0.01). In addition, the same treatment exhibited the highest total anthocyanin content (leaves; 36.1 × 10−2 ± 0.034 mg/g DW and root extract; 8.9 × 10−2 ± 0.020 mg/g DW), total phenolic content (stem extract; 4930.956 ± 16.025 mg GAE/g DE), and total flavonoid content (stem extract; 209.984 ± 0.572 mg QE/g DE). Moreover, this study also found that the highest antioxidant potential against 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,2-Azinobis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radicals was exhibited by samples supplemented with food waste compost (T3), followed by palm kernel biochar (T2). This indicates that the soil amendments have the capacity to enhance the secondary metabolites that protect plants, therefore ameliorating Melastoma malabathricum L.’s response towards acidic stress, and resulting in better antioxidant properties. Furthermore, this study also recorded better nutrient uptake in T3. With the significantly higher levels of macronutrient in the soil, the food waste compost could enhance the nutrient properties, secondary metabolites, and antioxidant capacity of Melastoma malabathricum L. grown in acidic soil conditions.
Study on the Role of Phytohormones in Resistance to Watermelon Fusarium Wilt
Authors: Zhu Feiying , Wang Zhiwei , Fang Yong , Tong Jianhua , Xiang Jing , Yang Kankan , Wang Ruozhong
Published: January 2022

Fusarium wilt disease is one of the major diseases causing a decline in watermelon yield and quality. Researches have informed that phytohormones play essential roles in regulating plants growth, development, and stress defendants. However, the molecular mechanism of salicylic acid (SA), jasmonic acid (JA), and abscisic acid (ABA) in resistance to watermelon Fusarium wilt remains unknown. In this experiment, we established the SA, JA, and ABA determination system in watermelon roots, and analyzed their roles in against watermelon Fusarium wilt compared to the resistant and susceptible varieties using transcriptome sequencing and RT-qPCR. Our results revealed that the up-regulated expression of Cla97C09G174770, Cla97C05G089520, Cla97C05G081210, Cla97C04G071000, and Cla97C10G198890 genes in resistant variety were key factors against (Fusarium oxysporum f. sp. Niveum) FON infection at 7 dpi. Additionally, there might be crosstalk between SA, JA, and ABA, caused by those differentially expressed (non-pathogen-related) NPRs, (Jasmonate-resistant) JAR, and (Pyrabactin resistance 1-like) PYLs genes, to trigger the plant immune system against FON infection. Overall, our results provide a theoretical basis for watermelon resistance breeding, in which phytohormones participate.
The Physiological Impact of GFLV Virus Infection on Grapevine Water Status: First Observations
Authors: Jež-Krebelj Anastazija , Rupnik-Cigoj Maja , Stele Marija , Chersicola Marko , Pompe-Novak Maruša , Sivilotti Paolo
Published: January 2022

In a vineyard, grapevines are simultaneously exposed to combinations of several abiotic (drought, extreme temperatures, salinity) and biotic stresses (phytoplasmas, viruses, bacteria). With climate change, the incidences of drought in vine growing regions are increased and the host range of pathogens with increased chances of virulent strain development has expanded. Therefore, we studied the impact of the combination of abiotic (drought) and biotic (Grapevine fanleaf virus (GFLV) infection) stress on physiological and molecular responses on the grapevine of cv. Schioppettino by studying the influence of drought and GFLV infection on plant water status of grapevines, on grapevine xylem vessel occlusion, and on expression patterns of 9-cis-epoxycarotenoid dioxygenase 1 (NCED1), 9-cis-epoxycarotenoid dioxygenase 2 (NCED2), WRKY encoding transcription factor (WRKY54) and RD22-like protein (RD22) genes in grapevines. A complex response of grapevine to the combination of drought and GFLV infection was shown, including priming in the case of grapevine water status, net effect in the case of area of occluded vessels in xylem, and different types of interaction of both stresses in the case of expression of four abscisic acid-related genes. Our results showed that mild (but not severe) water stress can be better sustained by GFLV infection rather than by healthy vines. GFLV proved to improve the resilience of the plants to water stress, which is an important outcome to cope with the challenges of global warming.
Mutational Analysis of a Wheat O-methyltransferase Involved in Flavonoid Metabolism
Authors: Cain Alexander B. , Yu Shu , Tian Li
Published: January 2022

Methylated flavones, and tricin in particular, have been implicated in protecting wheat plants against a variety of biotic and abiotic stresses. Methylated flavones are produced via O-methylation of the hydroxyl groups in flavones, which is catalyzed by O-methyltransferases (OMTs). To examine the role of wheat OMT2 in methylated flavone biosynthesis and facilitate interrogation of tricin functions in wheat-environment interactions, loss-of-function mutants of OMT2 homoeologs, omt-A2 and omt-B2, were identified from a tetraploid wheat Targeting Induced Local Lesions in Genomes (TILLING) mutant population and crossed to generate the omt-A2omt-B2 double mutant. Although tricin and most other soluble phenolics did not differ in leaves and glumes of TILLING control and the omt-A2, omt-B2, and omt-A2 omt-B2 mutants, chlorogenic acid was increased in glumes of omt-A2 omt-B2 relative to TILLING control, suggesting that it might serve as a substrate for OMT2. The omt2 mutant lines showed similar growth phenotypes as well as comparable lignin deposition in cell walls of stems compared to TILLING control. These results collectively suggest that OMT2 and its close homolog OMT1 may possess overlapping activities in tricin production, with OMT1 compensating for the missing OMT2 activities in the omt2 mutant lines.