bluuebunny/crossref_metadata_2025 huggingface dataset:
The Parasite Reduction Ratio (PRR) Assay Version 2: Standardized Assessment of Plasmodium falciparum Viability after Antimalarial Treatment In Vitro
Authors: Walz Annabelle , Duffey Maëlle , Aljayyoussi Ghaith , Sax Sibylle , Leroy Didier , Besson Dominique , Burrows Jeremy N. , Cherkaoui-Rbati Mohammed H. , Gobeau Nathalie , Westwood Marie-Anne , Siethoff Christoph , Gamo Francisco-Javier , Mäser Pascal , Wittlin Sergio
Published: January 2023

With artemisinin-resistant Plasmodium falciparum parasites emerging in Africa, the need for new antimalarial chemotypes is persistently high. The ideal pharmacodynamic parameters of a candidate drug are a rapid onset of action and a fast rate of parasite killing or clearance. To determine these parameters, it is essential to discriminate viable from nonviable parasites, which is complicated by the fact that viable parasites can be metabolically inactive, whilst dying parasites can still be metabolically active and morphologically unaffected. Standard growth inhibition assays, read out via microscopy or [3H] hypoxanthine incorporation, cannot reliably discriminate between viable and nonviable parasites. Conversely, the in vitro parasite reduction ratio (PRR) assay is able to measure viable parasites with high sensitivity. It provides valuable pharmacodynamic parameters, such as PRR, 99.9% parasite clearance time (PCT99.9%) and lag phase. Here we report the development of the PRR assay version 2 (V2), which comes with a shorter assay duration, optimized quality controls and an objective, automated analysis pipeline that systematically estimates PRR, PCT99.9% and lag time and returns meaningful secondary parameters such as the maximal killing rate of a drug (Emax) at the assayed concentration. These parameters can be fed directly into pharmacokinetic/pharmacodynamic models, hence aiding and standardizing lead selection, optimization, and dose prediction.
“Dual Anta-Inhibitors” of the A2A Adenosine Receptor and Casein Kinase CK1delta: Synthesis, Biological Evaluation, and Molecular Modeling Studies
Authors: Spinaci Andrea , Buccioni Michela , Catarzi Daniela , Cui Chang , Colotta Vittoria , Dal Ben Diego , Cescon Eleonora , Francucci Beatrice , Grieco Ilenia , Lambertucci Catia , Marucci Gabriella , Bassani Davide , Pavan Matteo , Varano Flavia , Federico Stephanie , Spalluto Giampiero , Moro Stefano , Volpini Rosaria
Published: January 2023

Based on a screening of a chemical library of A2A adenosine receptor (AR) antagonists, a series of di- and tri-substituted adenine derivatives were synthesized and tested for their ability to inhibit the activity of the enzyme casein kinase 1 delta (CK1δ) and to bind adenosine receptors (ARs). Some derivatives, here called “dual anta-inhibitors”, demonstrated good CK1δ inhibitory activity combined with a high binding affinity, especially for the A2AAR. The N6-methyl-(2-benzimidazolyl)-2-dimethyamino-9-cyclopentyladenine (17, IC50 = 0.59 μM and KiA2A = 0.076 μM) showed the best balance of A2AAR affinity and CK1δ inhibitory activity. Computational studies were performed to simulate, at the molecular level, the protein–ligand interactions involving the compounds of our series. Hence, the dual anta-inhibitor 17 could be considered the lead compound of new therapeutic agents endowed with synergistic effects for the treatment of chronic neurodegenerative and cancer diseases.
Arginine-Coated Nanoglobules for the Nasal Delivery of Insulin
Authors: Das Atanu , Vartak Richa , Islam Md Asrarul , Kumar Sunil , Shao Jun , Patel Ketan
Published: January 2023

Multiple daily injections via subcutaneous route are the primary modes of insulin delivery for patients with Diabetes Mellitus. While this process is invasive, painful and may cause patients to develop lipohypertrophy at injection site, the perception of fear surrounding this process causes patients to delay in initiation and remain persistent with insulin therapy over time. Moreover, poor glycemic control may often lead to acute complications, such as severe hypoglycemia and nocturnal hypoglycemia, especially in older patients with diabetes. To address the imperative need for a patient-convenient non-invasive insulin therapy, an insulin-loaded arginine-coated self-emulsifying nanoglobule system (INS-LANano) was developed for nasal delivery of insulin with a biodegradable cationic surfactant—Lauroyl Ethyl Arginate (LAE). Incorporation of LAE resulted in formation of positively charged nanoglobules with L-arginine oriented on the surface. LANano enabled binding of insulin molecules on the surface of nanoglobules via an electrostatic interaction between negatively charged α-helix and LAE molecules at physiological pH. INS-LANano showed a hydrodynamic diameter of 23.38 nm with a surface charge of +0.118 mV. The binding efficiency of insulin on LANano globules was confirmed by zeta potential, circular dichroism (CD) spectroscopy and centrifugal ultrafiltration studies. The attachment of insulin with permeation-enhancing nanoglobules demonstrated significantly higher in vitro permeability of insulin of 15.2% compared to insulin solution across human airway epithelial cell (Calu-3) monolayer. Upon intranasal administration of INS-LANano to diabetic rats at 2 IU/kg insulin dose, a rapid absorption of insulin with significantly higher Cmax of 14.3 mU/L and relative bioavailability (BA) of 23.3% was observed. Therefore, the INS-LANano formulation significant translational potential for intranasal delivery of insulin
Investigating the Effects of Olaparib on the Susceptibility of Glioblastoma Multiforme Tumour Cells to Natural Killer Cell-Mediated Responses
Authors: Moran Jennifer , Mylod Eimear , Kane Laura E. , Marion Caroline , Keenan Emily , Mekhaeil Marianna , Lysaght Joanne , Dev Kumlesh K. , O’Sullivan Jacintha , Conroy Melissa J.
Published: January 2023

Glioblastoma multiforme (GBM) is the most common adult primary brain malignancy, with dismal survival rates of ~14.6 months. The current standard-of-care consists of surgical resection and chemoradiotherapy, however the treatment response is limited by factors such as tumour heterogeneity, treatment resistance, the blood–brain barrier, and immunosuppression. Several immunotherapies have undergone clinical development for GBM but demonstrated inadequate efficacy, yet future combinatorial approaches are likely to hold more promise. Olaparib is FDA-approved for BRCA-mutated advanced ovarian and breast cancer, and clinical studies have revealed its utility as a safe and efficacious radio- and chemo-sensitiser in GBM. The ability of Olaparib to enhance natural killer (NK) cell-mediated responses has been reported in prostate, breast, and lung cancer. This study examined its potential combination with NK cell therapies in GBM by firstly investigating the susceptibility of the GBM cell line T98G to NK cells and, secondly, examining whether Olaparib can sensitise T98G cells to NK cell-mediated responses. Here, we characterise the NK receptor ligand profile of T98G cells and demonstrate that Olaparib does not dampen T98G susceptibility to NK cells or elicit immunomodulatory effects on the function of NK cells. This study provides novel insights into the potential combination of Olaparib with NK cell therapies for GBM.
Design and Characterization of Lipid-Surfactant-Based Systems for Enhancing Topical Anti-Inflammatory Activity of Ursolic Acid
Authors: Fonseca-Santos Bruno , Araujo Giovanna Angeli , Ferreira Paula Scanavez , Victorelli Francesca Damiani , Pironi Andressa Maria , Araújo Victor Hugo Sousa , Carvalho Suzana Gonçalves , Chorilli Marlus
Published: January 2023

Skin inflammation is a symptom of many skin diseases, such as eczema, psoriasis, and dermatitis, which cause rashes, redness, heat, or blistering. The use of natural products with anti-inflammatory properties has gained importance in treating these symptoms. Ursolic acid (UA), a promising natural compound that is used to treat skin diseases, exhibits low aqueous solubility, resulting in poor absorption and low bioavailability. Designing topical formulations focuses on providing adequate delivery via application to the skin surface. The aim of this study was to formulate and characterize lipid-surfactant-based systems for the delivery of UA. Microemulsions and liquid crystalline systems (LCs) were characterized by polarized light microscopy (PLM), rheology techniques, and textural and bioadhesive assays. PLM supported the self-assembly of these systems and elucidated their formation. Rheologic examination revealed pseudoplastic and thixotropic behavior appropriate, and assays confirmed the ability of these formulations to adhere to the skin. In vivo studies were performed, and inflammation induced by croton oil was assessed for response to microemulsions and LCs. UA anti-inflammatory activities of ~60% and 50% were demonstrated by two microemulsions and 40% and 35% by two LCs, respectively. These data support the continued development of colloidal systems to deliver UA to ameliorate skin inflammation.
Comparative Analysis of Dasatinib Effect between 2D and 3D Tumor Cell Cultures
Authors: Sabetta Samantha , Vecchiotti Davide , Clementi Letizia , Di Vito Nolfi Mauro , Zazzeroni Francesca , Angelucci Adriano
Published: January 2023

Three-dimensional cell culture methods are able to confer new predictive relevance to in vitro tumor models. In particular, the 3D multicellular tumor spheroids model is considered to better resemble tumor complexity associated with drug resistance compared to the 2D monolayer model. Recent advances in 3D printing techniques and suitable biomaterials have offered new promises in developing 3D tissue cultures at increased reproducibility and with high-throughput characteristics. In our study, we compared the sensitivity to dasatinib treatment in two different cancer cell lines, prostate cancer cells DU145 and glioblastoma cells U87, cultured in the 3D spheroids model and in the 3D bioprinting model. DU145 and U87 cells were able to proliferate in 3D alginate/gelatin bioprinted structures for two weeks, forming spheroid aggregates. The treatment with dasatinib demonstrated that bioprinted cells were considerably more resistant to drug toxicity than corresponding cells cultured in monolayer, in a way that was comparable to behavior observed in the 3D spheroids model. Recovery and analysis of cells from 3D bioprinted structures led us to hypothesize that dasatinib resistance was dependent on a scarce penetrance of the drug, a phenomenon commonly reported also in spheroids. In conclusion, the 3D bioprinted model utilizing alginate/gelatin hydrogel was demonstrated to be a suitable model in drug screening when spheroid growth is required, offering advantages in feasibility, reproducibility, and scalability compared to the classical 3D spheroids model.
Hyaluronan-Cyclodextrin Conjugates as Doxorubicin Delivery Systems
Authors: Bognanni Noemi , Viale Maurizio , La Piana Luana , Strano Simone , Gangemi Rosaria , Lombardo Cinzia , Cambria Maria Teresa , Vecchio Graziella
Published: January 2023

In the last years, nanoparticles based on cyclodextrins have been widely investigated for the delivery of anticancer drugs. In this work, we synthesized nanoparticles with a hyaluronic acid backbone functionalized with cyclodextrins under green conditions. We functionalized hyaluronic acid with two different molecular weights (about 11 kDa and 45 kDa) to compare their behavior as doxorubicin delivery systems. We found that the new hyaluronan-cyclodextrin conjugates increased the water solubility of doxorubicin. Moreover, we tested the antiproliferative activity of doxorubicin in the presence of the new cyclodextrin polymers in SK-N-SH and SK-N-SH-PMA (over-expressing CD44 receptor) cancer cells. We found that hyaluronan-cyclodextrin conjugates improved the uptake and antiproliferative activity of doxorubicin in the SK-N-SH-PMA compared to the SK-N-SH cell line at the ratio 8/1 doxorubicin/polymer. Notably, the system based on hyaluronan (45 kDa) was more effective as a drug carrier and significantly reduced the IC50 value of doxorubicin by about 56%. We also found that hyaluronic acid polymers determined an improved antiproliferative activity of doxorubicin (IC50 values are on average reduced by about 70% of free DOXO) in both cell lines at the ratio 16/1 doxorubicin/polymer.
Improving the Bioactivity of Norfloxacin with Tablets Made from Paper
Authors: Abdelkader Ayat , Nallbati Laura , Keck Cornelia M.
Published: January 2023

(1) Background: Many drugs possess poor bioavailability, and many strategies are available to overcome this issue. In this study, smartFilm technology, i.e., a porous cellulose matrix (paper), in which the active compound can be loaded onto in an amorphous state was utilised for oral administration to improve the solubility and bioactivity of a poorly soluble BSC class IV antibiotic. (2) Methods: Norfloxacin was used as the model drug and loaded into commercially available paper. The resulting norfloxacin-loaded smartFilms were transformed into smartFilm granules via wet granulation and the resulting norfloxacin-loaded smartFilm granules were transformed into norfloxacin-loaded tablets made from paper, i.e., smartFilm tablets. The crystalline state of norfloxacin was investigated, as well as the pharmaceutical properties of the granules and the tablets. The bioactivity of the smartFilm tablets was assessed in vitro and ex vivo to determine the antibacterial activity of norfloxacin. The results were compared to a physical mixture tablet that contained non-loaded paper granules and equal amounts of norfloxacin as a crystalline powder. (3) Results: Norfloxacin-loaded smartFilm granules and norfloxacin-loaded smartFilm tablets contained norfloxacin in an amorphous state, which resulted in an improved and faster release of norfloxacin when compared to the physical mixture tablet. The bioactivity was up to three times higher when compared to the physical mixture tablet. The ex vivo model was demonstrated to be a useful tool that allows for a fast and cost-effective discrimination between “good” and “bad” formulations. It provides realistic physiological conditions and can therefore yield meaningful, additional biopharmaceutical information that cannot be assessed in classical in vitro experiments. (4) Conclusions: smartFilm tablets are a promising, universal, industrially feasible and cost-effective formulation strategy for improved solubility and enhanced bioactivity of poorly soluble drugs.
Development of Nanoemulsions for Topical Application of Mupirocin
Authors: Alhasso Bahjat , Ghori Muhammad Usman , Conway Barbara R.
Published: January 2023

Mupirocin (MUP) is a topical antibacterial agent used to treat superficial skin infections but has limited application due to in vivo inactivation and plasma protein binding. A nanoemulsion formulation has the potential to enhance the delivery of mupirocin into the skin. MUP-loaded nanoemulsions were prepared using eucalyptus oil (EO) or eucalyptol (EU), Tween® 80 (T80) and Span® 80 (S80) as oil phase (O), surfactant (S) and cosurfactant (CoS). The nanoemulsions were characterised and their potential to enhance delivery was assessed using an in vitro skin model. Optimised nanoemulsion formulations were prepared based on EO (MUP-NE EO) and EU (MUP-NE EU) separately. MUP-NE EO had a smaller size with mean droplet diameter of 35.89 ± 0.68 nm and narrower particle size index (PDI) 0.10 ± 0.02 nm compared to MUP-NE EU. Both nanoemulsion formulations were stable at 25 °C for three months with the ability to enhance the transdermal permeation of MUP as compared to the control, Bactroban® cream. Inclusion of EU led to a two-fold increase in permeation of MUP compared to the control, while EO increased the percentage by 48% compared to the control. Additionally, more MUP was detected in the skin after 8 h following MUP-NE EU application, although MUP deposition from MUP-NE EO was higher after 24 h. It may be possible, through choice of essential oil to design nanoformulations for both acute and prophylactic management of topical infections.
Permeability Assessment of a High-Throughput Mucosal Platform
Authors: Butnarasu Cosmin , Garbero Olga Valentina , Petrini Paola , Visai Livia , Visentin Sonja
Published: January 2023

Permeability across cellular membranes is a key factor that influences absorption and distribution. Before absorption, many drugs must pass through the mucus barrier that covers all the wet surfaces of the human body. Cell-free in vitro tools currently used to evaluate permeability fail to effectively model the complexity of mucosal barriers. Here, we present an in vitro mucosal platform as a possible strategy for assessing permeability in a high-throughput setup. The PermeaPad 96-well plate was used as a permeability system and further coupled to a pathological, tridimensional mucus model. The physicochemical determinants predicting passive diffusion were determined by combining experimental and computational approaches. Drug solubility, size, and shape were found to be the critical properties governing permeability, while the charge of the drug was found to be influential on the interaction with mucus. Overall, the proposed mucosal platform could be a promising in vitro tool to model the complexity of mucosal tissues and could therefore be adopted for drug-permeability profiling.
Synthesis and Anti-Angiogenic Activity of Novel c(RGDyK) Peptide-Based JH-VII-139-1 Conjugates
Authors: Leonidis George , Koukiali Anastasia , Sigala Ioanna , Tsimaratou Katerina , Beis Dimitris , Giannakouros Thomas , Nikolakaki Eleni , Sarli Vasiliki
Published: January 2023

Peptide–drug conjugates are delivery systems for selective delivery of cytotoxic agents to target cancer cells. In this work, the optimized synthesis of JH-VII-139-1 and its c(RGDyK) peptide conjugates is presented. The low nanomolar SRPK1 inhibitor, JH-VII-139-1, which is an analogue of Alectinib, was linked to the ανβ3 targeting oligopeptide c(RGDyK) through amide, carbamate and urea linkers. The chemostability, cytotoxic and antiangiogenic properties of the synthesized hybrids were thoroughly studied. All conjugates retained mid nanomolar-level inhibitory activity against SRPK1 kinase and two out of four conjugates, geo75 and geo77 exhibited antiproliferative effects with low micromolar IC50 values against HeLa, K562, MDA-MB231 and MCF7 cancer cells. The activities were strongly related to the stability of the linkers and the release of JH-VII-139-1. In vivo zebrafish screening assays demonstrated the ability of the synthesized conjugates to inhibit the length or width of intersegmental vessels (ISVs). Flow cytometry experiments were used to test the cellular uptake of a fluorescein tagged hybrid in MCF7 and MDA-MB231 cells that revealed a receptor-mediated endocytosis process. In conclusion, most conjugates retained the inhibitory potency against SRPK1 as JH-VII-139-1 and demonstrated antiproliferative and antiangiogenic activities. Further animal model experiments are needed to uncover the full potential of such peptide conjugates in cancer therapy and angiogenesis-related diseases.
The Formulation of Curcumin: 2-Hydroxypropyl-β-cyclodextrin Complex with Smart Hydrogel for Prolonged Release of Curcumin
Authors: Nikolić Ljubiša , Urošević Maja , Nikolić Vesna , Gajić Ivana , Dinić Ana , Miljković Vojkan , Rakić Srđan , Đokić Sanja , Kesić Jelena , Ilić-Stojanović Snežana , Nikolić Goran
Published: January 2023

Curcumin comes from the plant species Curcuma longa and shows numerous pharmacological activities. There are numerous curcumin formulations with gels or cyclodextrins in order to increase its solubility and bioavailability. This paper presents the formulation of complex of curcumin with 2-hydroxypropyl-β-cyclodextrin in a thermosensitive hydrogel, based on N-isopropylmethacrylamide and N-isopropylacrylamide with ethylene glycol dimethacrylate as a crosslinker. The product was characterized by chemical methods and also by FTIR, HPLC, DSC, SEM, XRD. The results show that synthesis was successfully done. With an increase in the quantity of crosslinker in the hydrogels, the starting release and the release rate of curcumin from the formulation of the complex with hydrogels decreases. The release rate of curcumin from the gel complex formulation is constant over time. It is possible to design a formulation that will release curcumin for more than 60 days. In order to determine the mechanism and kinetics of curcumin release, various mathematical models were applied by using the DDSolver package for Microsoft Excel application. The Korsmeyer-Peppas model best describes the release of curcumin from the gel formulation of the complex, while the values for the diffusion exponent (0.063–0.074) shows that mechanism of the release rate is based on diffusion.
Silk Fibroin Bioink for 3D Printing in Tissue Regeneration: Controlled Release of MSC extracellular Vesicles
Authors: Bari Elia , Di Gravina Giulia Maria , Scocozza Franca , Perteghella Sara , Frongia Benedetta , Tengattini Sara , Segale Lorena , Torre Maria Luisa , Conti Michele
Published: January 2023

Sodium alginate (SA)-based hydrogels are often employed as bioink for three-dimensional (3D) scaffold bioprinting. They offer a suitable environment for cell proliferation and differentiation during tissue regeneration and also control the release of growth factors and mesenchymal stem cell secretome, which is useful for scaffold biointegration. However, such hydrogels show poor mechanical properties, fast-release kinetics, and low biological performance, hampering their successful clinical application. In this work, silk fibroin (SF), a protein with excellent biomechanical properties frequently used for controlled drug release, was blended with SA to obtain improved bioink and scaffold properties. Firstly, we produced a printable SA solution containing SF capable of the conformational change from Silk I (random coil) to Silk II (β-sheet): this transition is a fundamental condition to improve the scaffold’s mechanical properties. Then, the SA-SF blends’ printability and shape fidelity were demonstrated, and mechanical characterization of the printed hydrogels was performed: SF significantly increased compressive elastic modulus, while no influence on tensile response was detected. Finally, the release profile of Lyosecretome—a freeze-dried formulation of MSC-secretome containing extracellular vesicles (EV)—from scaffolds was determined: SF not only dramatically slowed the EV release rate, but also modified the kinetics and mechanism release with respect to the baseline of SA hydrogel. Overall, these results lay the foundation for the development of SA-SF bioinks with modulable mechanical and EV-release properties, and their application in 3D scaffold printing.
Poly(Glycerol) Microparticles as Drug Delivery Vehicle for Biomedical Use
Authors: Sahiner Mehtap , Yilmaz Aynur S. , Ayyala Ramesh S. , Sahiner Nurettin
Published: January 2023

Glycerol (Gly) is a well-known, FDA-approved molecule posing three hydroxyl groups. Since Gly is biocompatible, here, it was aimed to prepare poly(Glycerol) (p(Gly)) particles directly for the first time for the delivery of therapeutic agents. Micrometer-sized particles of p(Gly) were successfully synthesized via the micro-emulsion method with an average size of 14.5 ± 5.6 µm. P(Gly) microparticles up to 1.0 g/mL concentrations were found biocompatible with 85 ± 1% cell viability against L929 fibroblasts. Moreover, p(Gly) microparticles were tested for hemocompatibility, and it was found that up to 1.0 mg/mL concentrations the particles were non-hemolytic with 0.4 ± 0.1% hemolysis ratios. In addition, the blood compatibility index values of the prepared p(Gly) particles were found as 95 ± 2%, indicating that these microparticles are both bio- and hemocompatible. Furthermore, Quercetin (QC) flavonoid, which possessed high antioxidant properties, was loaded into p(Gly) microparticles to demonstrate drug-carrying properties of the particles with improved bioavailability, non-toxicity, and high biocompatibility. The results of this study evidently revealed that p(Gly) particles can be directly prepared from a cost-effective and easily accessible glycerol molecule and the prepared particles exhibited good biocompatibility, hemocompatibility, and non-toxicity. Therefore, p(Gly) particles were found as promising vehicles for drug delivery systems in terms of their higher loading and release capability as well as for sustained long term release profiles.
Productive, Morpho-Physiological, and Postharvest Performance of Six Basil Types Grown in a Floating Raft System: A Comparative Study
Authors: Ciriello Michele , Cirillo Valerio , Formisano Luigi , El-Nakhel Christophe , Pannico Antonio , De Pascale Stefania , Rouphael Youssef
Published: January 2023

Basil (Ocimum sp.) is one of the world’s most famous culinary fresh herbs, characterized by rapid growth that makes it particularly suitable for hydroponic cultivation. This study aimed to evaluate the adaptability of six types of basil to a closed-loop hydroponic system (floating raft system) and their post-harvest performance. Twenty-three days after transplantation, productivity, morpho-physiological performance, and mineral profile (by ion chromatography) were evaluated. At 3, 6, and 9 days after harvest, the loss of water from the from leaves stored at 10 °C in the dark was evaluated. Although the total fresh production of Thai, Mexican, and Genovese did not differ significantly, the latter provided a higher fresh leaf weight (16.52 g of plant−1) despite a lower leaf number (30.06 n. of plant−1). Nine days after harvest, Thai and Mexican showed the lowest water loss. Although Mexican Purple had the lowest net CO2 assimilation, it accumulated the highest concentration of ascorbic acid (909.41 mg 100 g fw−1).
Micropropagation as a Tool for the Conservation of Autochthonous Sorbus Species of Czechia
Authors: Šedivá Jana , Velebil Jiří , Zahradník Daniel
Published: January 2023

Members of the genus Sorbus are the only endemic tree species that occur in Czechia. They are important components of endangered plant communities. Their natural regeneration is usually problematic because of their mode of reproduction and because they can survive in rare populations with small numbers of individuals. The aim of this study was to develop a successful micropropagation protocol for selected Sorbus species, of which two are endemic (S. gemella and S. omissa) and two are hybrid (S. × abscondita and S. × kitaibeliana). We found significant differences in shoot induction and rooting ability between the Sorbus species under study. With the exception of S. × abscondita, N6-benzyladenine had a significantly greater effect on shoot regeneration, both in terms of shoot number and total shoot length, than meta-topolin. Root induction was key to the successful micropropagation of the Sorbus species studied. Our results show that four Sorbus species can be successfully rooted under ex vitro conditions, without a rooting powder treatment in a steamed peat-perlite substrate. Auxin-untreated microcuttings of S. gemella, S. × kitaibeliana and S. omissa, but not S. × abscondita, rooted better than ones treated with indole-3-butyric acid. This is the first time a micropropagation protocol for S. omissa, S. × abscondita and S. × kitaibeliana has been published.
Detecting Tomato Leaf Curl New Delhi Virus Causing Ridge Gourd Yellow Mosaic Disease, and Other Begomoviruses by Antibody-Based Methods
Authors: Naganur Priya , Shankarappa Kodegandlu Subbanna , Mesta Raghavendra K. , Rao Chilakalapudi Durga , Venkataravanappa Venkataravanappa , Maruthi Midatharahally Narasegowda , Reddy Lakshminarayana Reddy C. Narasimha
Published: January 2023

The incidence and severity of begomovirus diseases have been increasing around the world recently, and the ridge gourd [Luffa acutangula (Roxb.) L.] is the latest example of a crop that has become highly susceptible to the outbreak of the tomato leaf curl New Delhi virus (ToLCNDV, genus Begomovirus) in India. Accurate diagnosis of causal agents is important in designing disease management strategies. In this study the coat protein (CP) gene from a ToLCNDV-Rg ridge gourd isolate was used to produce polyclonal antibodies (ToLCNDV-Rg-CP-PAb) in a rabbit. The antibodies successfully detected a 30.5 kDa ToLCNDV-Rg-CP in extracts of symptomatic ridge gourd leaf samples by several assays, such as Western Blotting (WB), Dot Immuno Binding Assay (DIBA), Direct Antigen Coating Enzyme Linked Immuno Sorbent Assay (DAC-ELISA), Immuno Capture Polymerase Chain Reaction (IC-PCR), and Immuno Capture Loop-Mediated Isothermal Amplification (IC-LAMP) assays. However, none of the negative samples tested positive in either of the detection methods. Among all the methods tested, the immunocapture assay, IC-LAMP, was the most sensitive in detecting ToLCNDV-Rg. Furthermore, antibodies generated in this study also detected other commonly occurring begomoviruses in South India, such as tomato leaf curl Palampur virus and squash leaf curl China virus in cucurbits. Together, ToLCNDV-Rg-CP-PAb can be used for detecting at least three species of begomoviruses infecting cucurbits. The obtained antibodies will contribute to monitoring disease outbreaks in multiple crops.
Physiological Plasticity as a Strategy to Cope with Harsh Climatic Conditions: Ecophysiological Meta-Analysis of the Cosmopolitan Moss Ceratodon purpureus in the Southern Hemisphere
Authors: Beltrán-Sanz Núria , Raggio José , Pintado Ana , Dal Grande Francesco , García Sancho Leopoldo
Published: January 2023

Determining the physiological tolerance ranges of species is necessary to comprehend the limits of their responsiveness under strong abiotic pressures. For this purpose, the cosmopolitan moss Ceratodon purpureus (Hedw.) Brid. is a good model due to its wide geographical distribution throughout different biomes and habitats. In order to disentangle how this species copes with stresses such as extreme temperatures and high radiation, we designed a meta-analysis by including the main photosynthetic traits obtained by gas exchange measurements in three contrasting habitats from the Southern Hemisphere. Our findings highlight that traits such as respiration homeostasis, modulation of the photosynthetic efficiency, adjustment of the optimal temperature, and switching between shade and sun-adapted forms, which are crucial in determining the responsiveness of this species. In fact, these ecophysiological traits are in concordance with the climatic particularities of each habitat. Furthermore, the photosynthetic trends found in our study point out how different Livingston Island (Maritime Antarctica) and Granite Harbour (Continental Antarctica) are for plant life, while the population from the Succulent Karoo Desert (South Africa) shares traits with both Antarctic regions. Altogether, the study highlights the high resilience of C. purpureus under abrupt climate changes and opens new perspectives about the wide spectrum of physiological responses of cryptogams to cope with climate change scenarios.
Retrieval of Leaf Chlorophyll Contents (LCCs) in Litchi Based on Fractional Order Derivatives and VCPA-GA-ML Algorithms
Authors: Hasan Umut , Jia Kai , Wang Li , Wang Chongyang , Shen Ziqi , Yu Wenjie , Sun Yishan , Jiang Hao , Zhang Zhicong , Guo Jinfeng , Wang Jingzhe , Li Dan
Published: January 2023

The accurate estimation of leaf chlorophyll content (LCC) is a significant foundation in assessing litchi photosynthetic activity and possible nutrient status. Hyperspectral remote sensing data have been widely used in agricultural quantitative monitoring research for the non-destructive assessment of LCC. Variable selection approaches are crucial for analyzing high-dimensional datasets due to the high danger of overfitting, time-intensiveness, or substantial computational requirements. In this study, the performance of five machine learning regression algorithms (MLRAs) was investigated based on the hyperspectral fractional order derivative (FOD) reflection of 298 leaves together with the variable combination population analysis (VCPA)-genetic algorithm (GA) hybrid strategy in estimating the LCC of Litchi. The results showed that the correlation coefficient (r) between the 0.8-order derivative spectrum and LCC had the highest correlation coefficients (r = 0.9179, p < 0.01). The VCPA-GA hybrid strategy fully utilizes VCPA and GA while compensating for their limitations based on a large number of variables. Moreover, the model was developed using the selected 14 sensitive bands from 0.8-order hyperspectral reflectance data with the lowest root mean square error in prediction (RMSEP = 5.04 μg·cm−2). Compared with the five MLRAs, validation results confirmed that the ridge regression (RR) algorithm derived from the 0.2 order was the most effective for estimating the LCC with the coefficient of determination (R2 = 0.88), mean absolute error (MAE = 3.40 μg·cm−2), root mean square error (RMSE = 4.23 μg·cm−2), and ratio of performance to inter-quartile distance (RPIQ = 3.59). This study indicates that a hybrid variable selection strategy (VCPA-GA) and MLRAs are very effective in retrieving the LCC through hyperspectral reflectance at the leaf scale. The proposed methods could further provide some scientific basis for the hyperspectral remote sensing band setting of different platforms, such as an unmanned aerial vehicle (UAV) and satellite.
5-Aminolevulinic Acid Induces Chromium [Cr(VI)] Tolerance in Tomatoes by Alleviating Oxidative Damage and Protecting Photosystem II: A Mechanistic Approach
Authors: Kaya Cengiz , Ugurlar Ferhat , Ashraf Muhammed , Alyemeni Mohammed Nasser , Moustakas Michael , Ahmad Parvaiz
Published: January 2023

Chromium [Cr(VI)] pollution is a major environmental risk, reducing crop yields. 5-Aminolevunic acid (5-ALA) considerably improves plant abiotic stress tolerance by inducing hydrogen peroxide (H2O2) and nitric oxide (NO) signalling. Our investigation aimed to uncover the mechanism of tomato tolerance to Cr(VI) toxicity through the foliar application of 5-ALA for three days, fifteen days before Cr treatment. Chromium alone decreased plant biomass and photosynthetic pigments, but increased oxidative stress markers, i.e., H2O2 and lipid peroxidation (as MDA equivalent). Electrolyte leakage (EL), NO, nitrate reductase (NR), phytochelatins (PCs), glutathione (GSH), and enzymatic and non-enzymatic antioxidants were also increased. Foliar application of 5-ALA before Cr treatment improved plant growth and photosynthetic pigments, diminished H2O2, MDA content, and EL, and resulted in additional enhancements of enzymatic and non-enzymatic antioxidants, NR activity, and NO synthesis. In Cr-treated tomato seedlings, 5-ALA enhanced GSH and PCs, which modulated Cr sequestration to make it nontoxic. 5-ALA-induced Cr tolerance was further enhanced by sodium nitroprusside (SNP), a NO donor. When sodium tungstate (ST), a NR inhibitor, was supplied together with 5-ALA to Cr-treated plants, it eliminated the beneficial effects of 5-ALA by decreasing NR activity and NO synthesis, while the addition of SNP inverted the adverse effects of ST. We conclude that the mechanism by which 5-ALA induced Cr tolerance in tomato seedlings is mediated by NR-generated NO. Thus, NR and NO are twin players, reducing Cr toxicity in tomato plants via antioxidant signalling cascades.
Determination of Major, Minor and Chiral Components as Quality and Authenticity Markers of Rosa damascena Oil by GC-FID
Authors: Raeber Justine , Favrod Sina , Steuer Christian
Published: January 2023

Rose oil is traditionally produced by the water distillation of Rosa damascena and is of high economic value due to the low essential oil yield. It is therefore a common target for adulteration, which can cause harm to consumers. Current standards for authenticity control only consider the analysis of major components and overlook minor quality markers as well as the enantiomeric ratio of terpenes, which have proven useful in originality determination. The aim of this study was the development of two analytical GC-FID methods for the analysis of 21 and 29 rose oil analytes including major, minor and chiral components on a DB-wax and BGB 178 30% CD (chiral) capillary column, respectively. The total run time for both methods was within 60 min. For all target analytes, the % bias at the lower and upper calibration range varied from −7.8 to 13.2% and −13.1 to 5.2% analysed on the DB-wax column and 0.5 to 13.3% and −6.9 to 7.0% analysed on the chiral column. The chiral analysis successfully separated the enantiomers (+/−)-camphene, (+/−)-rose oxide, (+/−)-linalool, (+/−)-citronellol and (+/−)-citronellyl acetate, as well as the diastereomers of citral and β-damascenone. Both methods were applied to the analysis of 10 authentic rose oil samples and the enantiomeric/diastereomeric ratios, as well as the content of major and minor components, were determined. The identity of the analysed components in the authentic samples was further confirmed by GC-MS.
Rootstocks Comparison in Grafted Watermelon under Water Deficit: Effects on the Fruit Quality and Yield
Authors: Morales Carolina , Riveros-Burgos Camilo , Espinoza Seguel Felipe , Maldonado Carlos , Mashilo Jacob , Pinto Catalina , Contreras-Soto Rodrigo Iván
Published: January 2023

Drought is widely recognized as one of the most significant agricultural constraints worldwide. A strategy to avoid the adverse effects of drought on crops is to cultivate high-yielding varieties by grafting them onto drought-tolerant rootstocks with a differentiated root system. Thus, the objective of this study was to evaluate fruit yield and quality, root system architecture, and water productivity of watermelon grafted onto Lagenaria siceraria rootstocks. To do so, a commercial watermelon cultivar “Santa Amelia” [Citrullus lanatus (Thunb.)] was grafted onto five L. siceraria rootstocks: ‘Illapel’, ‘Osorno’, ‘BG-48’, ‘GC’, and ‘Philippines’, which were grown under three irrigation treatments (100%, 75%, and 50% of evapotranspiration). The comparison of the L. siceraria rootstocks in the irrigation treatments demonstrated no significant effect on watermelon fruit quality parameters. The rootstocks ‘Illapel’, ‘Osorno’, and ‘GC’ significantly improved the fruit number and yield (total fruit weight) under water deficit. Similarly, ‘Illapel’, ‘Osorno’, and ‘GC’ consistently showed statistical differences for root system architecture traits compared to ‘BG-48’ and ‘Philippines’. Based on these results, we concluded that the used L. siceraria rootstocks did not affect the fruit yield and quality of grafted watermelon under water deficit. This study may help adjust the amount of applied water for watermelon production where L. siceraria rootstocks are utilized.
Antioxidant, Bacteriostatic and Preservative Effects of Extractable Condensed Tannins Isolated from Longan Pericarps and Seeds
Authors: Wang Mengli , Chen Ting , Wang Qin , Shi Yan
Published: January 2023

In the process of longan production and processing, a large amount of remnants is produced, such as dried longan pericarps and seeds, which have been reported to be rich in polyphenols but not effectively utilized. In this paper, the total phenolic contents in the remnants of longan pericarps and seeds were found to be 39.58 ± 3.54 and 69.53 ± 1.99 mg/g (DW), respectively, accounting for 60–80% of those in fresh samples. The contents of extractable condensed tannins (ECTs) in the remnants of longan pericarps and seeds were 19.25 ± 6.71 mg/g (DW) and 44.59 ± 2.05 mg/g (DW), respectively, accounting for 60–70% of the fresh samples. These data indicate that the polyphenols in the remnants of the sampled longan pericarps and seeds were effectively retained. The antioxidant capacity of ECTs from the longan pericarps and seeds was more than 60% of the fresh samples measured with the 1, 1-diphenyl-2-trinitrophenylhydrazine and ferric reducing ability of plasma methods. Further exploration showed that ECTs from the longan pericarps and seeds had significant inhibitory effects on Pseudomonas aeruginosa, Escherichia coli, Salmonella and Staphylococcus aureus. The minimum inhibitory concentration (MIC) of the longan pericarp ECTs on all four studied bacteria was 3 mg/mL. The MIC of longan seed ECTs on Salmonella was 3 mg/mL, and that of the other three bacteria was 1.5 mg/mL. In view of the good antioxidant and antibacterial activities of longan pericarps and seeds, we applied them to the preservation of fresh-cut lotus roots. When the concentration of ECTs in the longan pericarps and seeds was 2 mg/mL and 1 mg/mL, respectively, the two kinds of ECTs showed an obvious preservative effect. After the ECT treatment of the lotus roots, their browning degree was reduced, their color was better maintained, their respiration was inhibited and their nutrient loss was reduced. Bacterial reproduction was inhibited, and cell senescence was slowed. Accordingly, the shelf life of ECT-treated fruits and vegetables can be effectively extended. Overall, we can suggest that ECTs from the remnants of dried longan pericarps and seeds could be used as natural preservatives for fresh-cut fruits and vegetables.
Structural and Physiological Traits of Compound Leaves of Ceratonia siliqua Trees Grown in Urban and Suburban Ambient Conditions
Authors: Papadopoulou Sophia , Stefi Aikaterina L. , Meletiou-Christou Maria-Sonia , Christodoulakis Nikolaos S. , Gkikas Dimitrios , Rhizopoulou Sophia
Published: January 2023

Ceratonia siliqua L. (carob tree) is an endemic plant to the eastern Mediterranean region. In the present study, anatomical and physiological traits of successively grown compound leaves (i.e., the first, third, fifth and seventh leaves) of C. siliqua were investigated in an attempt to evaluate their growth under urban and suburban environmental conditions. Chlorophyll and phenolic content, as well as the specific leaf area of the compound leaves were determined. Structural traits of leaflets (i.e., thickness of palisade and spongy parenchyma, abaxial and adaxial epidermis, as well as abaxial and adaxial periclinal wall) were also investigated in expanding and fully expanded leaflets. Fully expanded leaflets from urban sites exhibited increased thickness of the lamina and the palisade parenchyma, while the thickness of the spongy parenchyma was thicker in suburban specimens. The palisade tissue was less extended than the spongy tissue in expanding leaflets, while the opposite held true for the expanded leaflets. Moreover, the thickness of the adaxial and the abaxial epidermises, as well as the adaxial and abaxial periclinal wall were higher in suburban leaflets. The chlorophyll content increased concomitantly with the specific leaf area (SLA) of both expanding and expanded leaflets, and strong positive correlations were detected, while the phenolic content declined with the increased SLA of expanding and expanded leaflets. It is noteworthy that the SLA of expanding leaflets in the suburban site was comparable to the SLA of expanded leaflets experiencing air pollution in urban sites; the size and the mass of leaf blades of C. siliqua possess adaptive features to air pollution. These results, linked to the functional structure of expanding and expanded successive foliar tissues, provide valuable assessment information coordinated with an adaptive process and yield of carob trees exposed to the considered ambient conditions, which have not hitherto been published.
Morphological and Genetic Clonal Diversity within the ‘Greco Bianco’ Grapevine (Vitis vinifera L.) Variety
Authors: Villano Clizia , Corrado Giandomenico , Basile Boris , Di Serio Ermanno , Mataffo Alessandro , Ferrara Elvira , Aversano Riccardo
Published: January 2023

Grapevine (Vitis vinifera L.) has been propagated vegetatively for hundreds of years. Therefore, plants tend to accumulate somatic mutations that can result in an intra-varietal diversity capable of generating distinct clones. Although it is common that winemakers request specific clones or selections for planting new vineyards, relatively limited information is available on the extent, degree, and morphological impact of the clonal diversity in traditional, highly valued grapevine varieties within production areas protected by geographical denomination of origin. Here, we present a morphological and genetic investigation of the intra-varietal diversity in ‘Greco Bianco’, the grapevine variety used to produce the DOCG and PDO “Greco di Tufo” wine. Seventeen clones from different farms (all within the allowed production area) were phenotypically characterized using ampelographic and ampelometric traits. The clones were also genotyped with Simple Sequence Repeats (SSR) and retrotransposon-based DNA markers (REMAP). The morphological analysis indicated a uniformity in the qualitatively scored traits, and a limited variability for the quantitative traits of the bunch and of the berry composition. The molecular markers also depicted variability among clones, which was more evident with the use of REMAPs. The comparison of the discriminatory information of the three analyses indicated that they provided different estimates of the level of diversity. The evaluation described herein of the clonal variability has implications for the management and protection of clonal selections in ‘Greco Bianco’ and prompts for further multidisciplinary investigations on its possible role in winemaking.
Friends in Arms: Flavonoids and the Auxin/Cytokinin Balance in Terrestrialization
Authors: Kurepa Jasmina , Shull Timothy E. , Smalle Jan A.
Published: January 2023

Land plants survive the challenges of new environments by evolving mechanisms that protect them from excess irradiation, nutrient deficiency, and temperature and water availability fluctuations. One such evolved mechanism is the regulation of the shoot/root growth ratio in response to water and nutrient availability by balancing the actions of the hormones auxin and cytokinin. Plant terrestrialization co-occurred with a dramatic expansion in secondary metabolism, particularly with the evolution and establishment of the flavonoid biosynthetic pathway. Flavonoid biosynthesis is responsive to a wide range of stresses, and the numerous synthesized flavonoid species offer two main evolutionary advantages to land plants. First, flavonoids are antioxidants and thus defend plants against those adverse conditions that lead to the overproduction of reactive oxygen species. Second, flavonoids aid in protecting plants against water and nutrient deficiency by modulating root development and establishing symbiotic relations with beneficial soil fungi and bacteria. Here, we review different aspects of the relationships between the auxin/cytokinin module and flavonoids. The current body of knowledge suggests that whereas both auxin and cytokinin regulate flavonoid biosynthesis, flavonoids act to fine-tune only auxin, which in turn regulates cytokinin action. This conclusion agrees with the established master regulatory function of auxin in controlling the shoot/root growth ratio.