Search

Infection with Plasmodium falciparum enhances extracellular vesicle (EV) production in parasitized red blood cells (pRBCs), an important mechanism for parasite-to-parasite communication during the asexual intraerythrocytic life cycle. The endosomal sorting complex required for transport (ESCRT), and in particular the ESCRT-III sub-complex, participates in the formation of EVs in higher eukaryotes. However, RBCs have lost the majority of their organelles through the maturation process, including an important reduction in their vesicular network. Therefore, the mechanism of EV production in P. falciparum-infected RBCs remains to be elucidated. Here we demonstrate that P. falciparum possesses a functional ESCRT-III machinery activated by an alternative recruitment pathway involving the action of PfBro1 and PfVps32/PfVps60 proteins. Additionally, multivesicular body formation and membrane shedding, both reported mechanisms of EVs production, were reconstituted in the membrane model of giant unilamellar vesicles using the purified recombinant proteins. Moreover, the presence of PfVps32, PfVps60 and PfBro1 in EVs purified from a pRBC culture was confirmed by super-resolution microscopy and dot blot assays. Finally, disruption of the Pfvps60 gene led to a reduction in the number of the produced EVs in the KO strain and affected the distribution of other ESCRT-III components. Overall, our results increase the knowledge on the underlying molecular mechanisms during malaria pathogenesis and demonstrate that ESCRT-III P. falciparum proteins participate in EV production. Author summary: Malaria is a disease caused by Plasmodium parasites that is still a leading cause of death in many low-income countries, and for which currently available therapeutic strategies are not succeeding in its control, let alone eradication. An interesting feature observed after Plasmodium invasion is the increase of extracellular vesicles (EVs) generated by parasitized red blood cells (pRBCs), which lack a vesicular trafficking that would explain EV production. Here, by combining different approaches, we demonstrated the participation of the endosomal sorting complex required for transport (ESCRT) machinery from Plasmodium falciparum in the production of EVs in pRBCs. Moreover, we were able to detect ESCRT-III proteins adjacent to the membrane of the host and in EVs purified from a pRBC culture, which shows the export of these proteins and their participation in EV production. Finally, the disruption of an ESCRT-III associated gene, Pfvps60, led to a significant reduction in the amount of EVs. Altogether, these results confirm ESCRT-III participation in EV production and provide novel information on the P. falciparum protein export mechanisms, which can be used for the development of new therapeutic strategies against malaria, based on the disruption of EV formation and trafficking. [ABSTRACT FROM AUTHOR]

Plants have been used as the main source of phytochemicals with nutritional, medicinal, cultural and cosmetic applications since times immemorial. Nowadays, achieving sustainable development, global climate change, restricted access to fresh water, limited food supply and growing energy demands are among the critical global challenges faced by humanity. Plant cell culture technology has the potential to address some of these challenges by providing effective tools for sustainable supply of phyto‐ingredients with reduced energy, carbon and water footprints. The main aim of this review is to discuss the recent trends in the development of plant cell culture technologies for production of plant‐derived substances with application in food products and cosmetic formulations. The specific technological steps and requirements for the final products are discussed in the light of the advances in cultivation technologies used for growing differentiated and undifferentiated plant in vitro systems. Future prospects and existing challenges of the commercialization of plant cell culture‐derived products have been outlined through the prism of the authors' point of view. We expect this review will encourage scientists, policymakers and business enterprises to join efforts for speeding‐up the mass commercialization and popularization of plant cell culture technology as an eco‐friendly alternative method for sustainable production of plant‐derived additives with application in food and cosmetic products. [ABSTRACT FROM AUTHOR]

Plants have always been the main source for active cosmetic ingredients, having proven health beneficial effects on human, such as anti‐aging, antioxidant, anti‐inflammatory, UV‐protective, anti‐cancer, anti‐wrinkle, skin soothing, whitening, moisturizing, etc. Extracts from herbal, aromatic and/or medicinal plants have been widely used as effective active ingredients in cosmeceuticals or nutricosmetics, especially in products for topical application and skin‐care formulations. However, over the past decade, there has been an increasing interest to plant cell culture – derived active cosmetic ingredients. These are "new generation" of high quality natural products, produced by the modern plan biotechnology methods, which usually showed stronger activities than the plant extracts obtained by the classical methods. In this review, the advantages and the current progress in plant cell culture technology for the production of active cosmetic ingredients have been summarized, and discussed in details within a presented case study for calendula stem cell product development. [ABSTRACT FROM AUTHOR]

Abstract: Biomembranes are constantly remodeled and in cells, these processes are controlled and modulated by an assortment of membrane proteins. Here, it is shown that such remodeling can also be induced by photoresponsive molecules. The morphological control of giant vesicles in the presence of a water‐soluble ortho‐tetrafluoroazobenzene photoswitch (F‐azo) is demonstrated and it is shown that the shape transformations are based on an increase in membrane area and generation of spontaneous curvature. The vesicles exhibit budding and the buds can be retracted by using light of a different wavelength. In the presence of F‐azo, the membrane area can increase by more than 5% as assessed from vesicle electrodeformation. To elucidate the underlying molecular mechanism and the partitioning of F‐azo in the membrane, molecular dynamics simulations are employed. Comparison with theoretically calculated shapes reveals that the budded shapes are governed by curvature elasticity, that the spontaneous curvature can be decomposed into a local and a nonlocal contribution, and that the local spontaneous curvature is about 1/(2.5 µm). The results show that exo‐ and endocytotic events can be controlled by light and that these photoinduced processes provide an attractive method to change membrane area and morphology. [ABSTRACT FROM AUTHOR]

After eight years of cultivation of Sea daffodil (Pancratium maritimum L.) shoot cultures at in vitro conditions, bacterial contamination on the plant tissues and medium surface appeared. The contamination was due to endophytic bacterium later isolated as a bacterial strain PM_YT. The colonial characteristics and cell morphology of the isolate were determined after cultivation on YEB-agar medium and coloring of microscopic preparations. The strain identification was implemented by biochemical methods (oxidase and catalase standard strip tests) and nucleotide sequencing of the 16S rRNA gene. The results from the morphological and cultural identification showed that the isolate was Gram-positive bacterium presented from small, short rods, forming small (size of 1-3 mm), round-shaped, yellow- to slightly orange-colored colonies. The results from biochemical identification showed that the strain was oxidase-negative and catalasepositive. The comparative 16S rRNA gene sequence-based phylogenetic analysis revealed 99% pairwise similarity of the strain PM_YT to the bacterial species Curtobacterium flaccumfaciens. Antibiotic susceptibility was implemented by Bauer-Kirby disc diffusion method and the results demonstrated that the isolate was most sensitive to the antibiotics belonging to inhibitors of protein synthesis group, especially to tetracycline, doxycycline, amikacin, rifampin, lincomycin, chloramphenicol, erythromycin, streptomycin and ciprofloxacin. [ABSTRACT FROM AUTHOR]

Plant tissue and organ cultures in vitro usually face technological challenges. When submerged cultivation of plant cells in a controlled environment is desired, the characteristic growth morphology and physiology of differentiated organ cultures present a problem in process scale-up. Temporary immersion systems (TIS) were developed several decades ago. These systems are providing the most natural environment for in vitro culture of plant shoots and seedlings. Over the past few years, TIS have been recognized as a perspective technology for plant micropropagation, production of plant-derived secondary metabolites, expression of foreign proteins, and potential solutions in phytoremediation. Nowadays, several TIS, operating on similar or divergent technological principles, have been developed and successfully applied in the cultivation of various plant in vitro systems, including somatic embryos and transformed root cultures. In this article, the operational principle and technological design of the most popular TIS are reviewed. In addition, recent examples of the application of temporary immersion technology for in vitro cultivation of plant tissue and organ cultures at laboratory and pilot scales are discussed. Finally, future prospects and challenges to the industrial realization of that fast-developing technique are outlined. [ABSTRACT FROM AUTHOR]

The constituents of essential oils, obtained from different aerial parts (leaves and flowers) of Salvia ringens Sibth. et Sm. growing wild in Bulgaria, have been analyzed by GC-MS. The yields of leaves and flowers oils were similar (0.03% v/w and 0.04% v/w). 60 compounds were identified in both oils, which correspond to 97.7% and 97.3% of the total oils constituents for leaves and flower oils, respectively. Both leaves and flowers oils showed similar chemical compositions, with predominance of camphor (17.2% and 18.8%) and borneol (7.2% and 8.7%). Further analyses of minor component distributions proves that the both leaves and flowers could be equally used as a raw material for obtaining of camphor reach essential oil from Salvia ringens Sibth. et Sm. [ABSTRACT FROM AUTHOR]

Abstract: The effects of methyl jasmonate and jasmonic acid on galanthamine production, phenolic acid content and growth of Leucojum aestivum L. shoot culture, cultivated in submerged conditions were investigated. The best time-point for addition of elicitors was during the exponential phase of the culture growth. The maximal contents of galanthamine and lycorine (226.9μg/flask and 491.4μg/flask, 1.36 and 1.67-fold higher compared to the control, respectively) were achieved after elicitation with jasmonic acid, whereas the elicitation with methyl jasmonte resulted in maximal accumulation of phenolic acids. It was demonstrated that the boosting effect of jasmonic acid on Amaryllidacea alkaloid biosynthesis was due to induction of the activity of tyrosine decarboxylase, whereas methyl jasmonate stimulates the biosynthesis of phenolic acids by inducing mainly the activity of phenylalanine ammonia-lyase. [Copyright &y& Elsevier]

Biomass accumulation and tropane alkaloids production by diploid and tetraploid hairy root cultures of Datura stramonium L. cultivated in stirred tank bioreactor at different aeration rates were investigated. The maximal growth for both hairy root cultures (ADB = 8.3 g/L and 6.8 g/L for diploid and tetraploid line, respectively) was achieved at aeration rate of 15.0 L/(L.h). The corresponding growth indexes were remarkably high (GIDW = 9.0 and 7.8 for diploid and tetraploid line, respectively) compared to the values, usually reported for other hairy root cultures. The optimal aeration rate for biomass accumulation was also optimal for alkaloids biosynthesis. According to our survey, the achieved maximal amounts of accumulated hyoscyamine (35.0 mg/L and 27.0 mg/L for diploid and tetraploid line) were the highest reported in the scientific literature for D. stramonium L. hairy roots. During the cultivation in stirred tank bioreactor, the hairy roots biosynthesized pharmaceutically important alkaloid scopolamine in minor concentrations. This is an important observation since scopolamine was not detected during submerged cultivation of these hairy root lines in other bioreactor types. However, the ploidy level was found to be the most important factor concerning scopolamine production by D. stramonium L. hairy roots cultures. The present work demonstrated the effect of ploidity levels on biomass accumulation and tropane alkaloids production by D. stramonium L. hairy roots cultivated in stirred tank bioreactor. This investigation show that the stirred tank bioreactor could be successfully applied for both maximal biomass accumulations, as well as for manipulation of tropane alkaloids production by diploid and tetraploid D. stramonium L. hairy root cultures. [ABSTRACT FROM AUTHOR]

Shoot culture of summer snowflake ( Leucojum aestivum L.) was successfully cultivated in an advanced modified glass-column bioreactor with internal sections for production of Amaryllidaceae alkaloids. The highest amounts of dry biomass (20.8 g/L) and galanthamine (1.7 mg/L) were achieved when shoots were cultured at 22°C and 18 L/(L·h) flow rate of inlet air. At these conditions, the L. aestivum shoot culture possessed mixotrophic-type nutrition, synthesizing the highest amounts of chlorophyll (0.24 mg/g DW (dry weight) chlorophyll A and 0.13 mg/g DW chlorophyll B). The alkaloids extract of shoot biomass showed high acetylcholinesterase inhibitory activity ( IC50 = 4.6 mg). The gas chromatography-mass spectrometry (GC/MS) profiling of biosynthesized alkaloids revealed that galanthamine and related compounds were presented in higher extracellular proportions while lycorine and hemanthamine-type compounds had higher intracellular proportions. The developed modified bubble-column bioreactor with internal sections provided conditions ensuring the growth and galanthamine production by L. aestivum shoot culture. [ABSTRACT FROM AUTHOR]

Abstract: The alkaloid patterns in Leucojum aestivum L. shoot culture cultivated at temporary immersion conditions were investigated using gas chromatography–mass spectrometry. 18 alkaloids were identified, and galanthamine, hamayne and lycorine were dominant. The L. aestivum 80 shoot culture, cultivated at temporary immersion conditions, is a prospective biological matrix for obtaining wide range Amaryllidaceae alkaloids, showing valuable biological and pharmacological activities. The temperature of cultivation influenced enzyme activities, catalyzing phenol oxidative coupling of 4′-O-methylnorbelladine and formation of the different groups Amaryllidaceae alkaloids. Decreasing the temperature of cultivation of L. aestivum 80 shoot culture led to activation of para-ortho’ phenol oxidative coupling (formation of galanthamine type alkaloids) and inhibited ortho-para’ and para-para’ phenol oxidative coupling (formation of lycorine and haemanthamine types alkaloids). [Copyright &y& Elsevier]

Abstract: Results of the semi-analytical modelling of the interaction of laser-initiated airborne shock waves with an infinite horizontally positioned elastic plate are presented. The impact of the airborne shock waves on the plate is approximated by a cylindrically diverging surface force resulting from the pressure of the incident and reflected shock waves. This force is then represented in the wavenumber–frequency domain by means of Hankel and Fourier transforms that are carried out numerically so that the interaction problem can be solved using the Green’s function method. The resulting frequency spectra and time histories of generated flexural wave pulses are calculated for different values of laser pulse energy and for different heights of the laser beam focusing above the plate surface. The theoretical results obtained are compared with the results of laboratory measurements of the interaction of laser-generated acoustic shocks with a large plastic plate. The comparison shows reasonably good agreement between the semi-analytical predictions and data. [ABSTRACT FROM AUTHOR]

Membrane fission triggered by the endosomal sorting complex required for transport (ESCRT) is an important process observed in several pathogenic and non-pathogenic cellular events. From a synthetic-biology viewpoint, ESCRT proteins represent an interesting machinery for the construction of cell mimetic sub-compartments produced by fission. Since their discovery, the studies on ESCRT-III-mediated action, have mainly focused on protein dynamics, ignoring the role of lipid organization and membrane phase state. Recently, it has been suggested that membrane buds formed by the action of ESCRT-III are generated from transient microdomains in endosomal membranes. However, the interplay between membrane domain formation and ESCRT remodeling pathways has not been investigated. Here, giant unilamellar vesicles made of ternary lipid mixtures, either homogeneous in phase or exhibiting liquid-ordered/liquid-disordered phase coexistence, were employed as a model membrane system. These vesicles were incubated with purified recombinant ESCRT-III proteins from the parasite Entamoeba histolytica. In homogeneous membranes, we observe that EhVps32 can trigger domain formation while EhVps20 preferentially co-localizes in the liquid disordered phase. The addition of EhVps24 appears to induce the formation of intraluminal vesicles produced from the liquid-ordered phase. In phase separated membranes, the intraluminal vesicles are also generated from the liquid-ordered phase and presumably emerge from the phase boundary region. Our findings reinforce the hypothesis that ESCRT-mediated remodeling depends on the membrane phase state. Furthermore, the obtained results point to a potential synthetic biology approach for establishing eukaryotic mimics of artificial cells with microcompartments of specific membrane composition, which can also differ from that of the mother vesicle. [Display omitted] • Binding of 3 ESCRT-III proteins to homogeneous and phase-separated GUVs is studied. • EhVps20t binds uniformly to homogeneous charged ternary-mixture membranes. • Subsequent EhVps32 binding induces fluid-fluid (Lo/Ld) phase-separation. • ESCRT-III proteins appear to cause Lo buds originating at the Lo/Ld phase boundary. [ABSTRACT FROM AUTHOR]

The growth and hyosciamine biosynthesis by diploid and tetraploid Datura stramonium L. hairy rootlines in temporary immersion RITA" system were investigated. The immersion frequency had significant effect on hyosciamine production and accumulation of biomass for both hairy root cultures. The maximum amount of hyosciamine (6.53 mg.g-1 DW) was achieved during cultivation of diploid Datura stramonium L. hairy root culture at immersion frequency with 15 rain flooding and 105 min stand-by periods. Data for utilization of the main inorganic compounds during cultivation process in temporary immersion RITA" system are presented as well. [ABSTRACT FROM AUTHOR]

Abstract: Various stressful paradigms were found to induce anticonvulsant effects in different seizure models. Methylxanthines, such as theophylline might contribute to the reduction of restraint-induced stress. Therefore, in this study the influence of acute restraint stress on pentylenetetrazol (PTZ) seizure thresholds as well as the effect of acute and chronic theophylline pretreatment on stress-induced modulation of the seizure threshold were assessed in mice. The onset of the three consecutive seizure phases: myoclonic twitch (MTW), generalized clonus (GNCL) and tonic hind limb extension (THE) was delayed after exposure to a 2h restraint stress by 34%, 23% and 24%, respectively. In nonstressed mice, acute theophylline injection (100mg/kg, i.p.) decreased the threshold only for THE. However, in stressed animals, the pretreatment with the methylxanthine significantly enhanced the dose of the convulsant producing the same seizure phase. In nonstressed mice, long-term theophylline treatment (50mg/kg, twice daily for 14 days) increased PTZ threshold for all three seizure phases. In contrast, in chronically treated with theophylline mice exposed to restraint stress, significant decrease in the PTZ threshold for all seizure phases compared to control stressed animals have been observed. These results suggest that, depending on the treatment regimen (acute versus chronic), theophylline specifically and differentially modulates the anticonvulsant effect of restraint stress in mice. [Copyright &y& Elsevier]

Abstract: Accumulated studies support the concept that angiotensin peptides, ANG II, ANG III, and ANG IV act as neurotransmitters or neuromodulators in specific neuronal pathways in the brain stem, the hypothalamus, and the forebrain. They have been implicated in the regulation of several physiological processes, particularly in excitable brain structures that express high concentration of their receptors. With the help of pharmacological approaches it was shown that angiotensin peptides appear to be anticonvulsant in a variety of experimental seizure models. Thus, ANG II increases the threshold for pentylenetetrazol (PTZ)-, bicuculline-and picrotoxin-induced seizures in mice. It also attenuates the intensity of clonic seizures evoked by PTZ and 3-mercaptopropionic acid and is effective in the maximal electroshock test. Furthermore, ANG II, ANG III, and ANG IV protect against the clonic convulsions in the PTZ kindling model of epilepsy in mice. From the accumulated results it could be assumed that the angiotensin peptides appear to realize their effects acting directly on their receptors (AT1, AT2 and AT4) and through close interaction with different neurotransmitter/neuromodulator systems as dopamine (DA)-, γ-aminobutyric acid (GABA)-and adenosine. This may contribute to a new potential use of angiotensin drugs either alone or in combination with other neuroprotective agents acting through the above mentioned systems, thus providing a more rational strategy for the treatment of neurodegenerative disorders such as epilepsy. [Copyright &y& Elsevier]

The effects of adenosine A1 and A2A receptor agonists and antagonists administered intraperitoneally (ip) and their interaction with angiotensin II (Ang II) administered intracerebroventricularly (icv) were studied in mice using the acetic acid-induced abdominal constriction test. Ang II (0.1 μg/mouse) induced antinociception in this model. The adenosine A1 receptor agonist N6-cyclopentyladenosine (CPA; 0.05, 0.25 and 0.5 mg/kg) also showed a well-developed antinociceptive effect. Ang II (0.1 μg/mouse) administered 5 min before CPA (0.25 mg/kg) decreased the number of writhes, i.e., it enhanced the antinociceptive effect of CPA. Losartan, an AT1 receptor antagonist (25 μg/mouse icv), enhanced the antinociceptive effect of CPA, while the AT2 receptor antagonist 1-[-4-(dimethylamino)-3-methylphenylmethyl]-5-diphenylacetyl)-4,5,6,7-tetrahydro 1H-4-imidazol [4,5c]pyridine-6 carboxylic acid, ditrifluoroacetate, dihydrate (PD 123319; 10 μg/mouse) had less effect. 8-Cyclopentyl-1,3-dipropylxanthine (DPCPX; 0.1 mg/kg), an adenosine A1 receptor antagonist, exhibited a pronociceptive effect and did not change the antinociceptive effect of Ang II. The adenosine A2A receptor agonist PD-125944 (DPMA; 0.1, 0.5 and 1 mg/kg) showed pronounced antinociceptive effect. Ang II (0.1 μg/mouse) did not significantly influence the antinociceptive effect of DPMA (0.1 mg/kg). The A2A receptor antagonist 3,7-dimethyl-1-propargilxanthine (DMPX; 0.1 mg/kg) had no effect on the number of writhes and did not influence the effect of Ang II. These data indicate that the antinociceptive effect of Ang II interacts with that produced by adenosine A1 receptor agonist. [Copyright &y& Elsevier]

DOI: 10.1002/elsc.202000078 The cover picture shows a process of submerged cultivation of hairy root culture of red beet in a stirred tank bioreactor, different types I Beta vulgaris i L. in vitro cultures and their ploidity levels. Among the investigated red beet in vitro cultures, hairy roots showed the best potential for production of betalains. [Extracted from the article]

Plants belonging to the monocotyledonous Amaryllidaceae family include about 1100 species divided among 75 genera. They are well known as medicinal and ornamental plants, producing pharmaceutically important alkaloids, the most intensively investigated of which are galanthamine and lycorine. Amaryllidaceae alkaloids possess various biological activities, the most important one being their anti-acetylcholinesterase activity, used for the treatment of Alzheimer's disease. Due to increased demand for Amaryllidaceae alkaloids (mainly galanthamine) and the limited availability of plant sources, in vitro culture technology has attracted the attention of researchers as a prospective alternative for their sustainable production. Plant in vitro systems have been extensively used for continuous, sustainable, and economically viable production of bioactive plant secondary metabolites. Over the past two decades, a significant success has been demonstrated in the development of in vitro systems synthesizing Amaryllidaceae alkaloids. The present review discusses the state of the art of in vitro Amaryllidaceae alkaloids production, summarizing recently documented plant in vitro systems producing them, as well as the authors' point of view on the development of biotechnological production processes with a focus on the future prospects of in vitro culture technology for the commercial production of these valuable alkaloids. [ABSTRACT FROM AUTHOR]

Abstract: Three often cited systems for the extraction of plant nuclei for flow cytometric measurement (CyStain PI, Partec GmbH, Münster, Germany, the method of Arumuganathan and Earle, and LB01 buffer) failed, when applied to the hairy roots of red beet (Beta vulgaris). By combining these systems and introducing a centrifugation step, the extraction, staining, and analysis of nuclei from this tissue were performed successfully. [Copyright &y& Elsevier]

Essential oils and hydrosols of two cultivars of muscadine grapes (Muscadinia rotundifolia (Michx.) Small.) were obtained by hydro-distillation of flowers and berry skins. Twenty-three volatile compounds were identified in essential oils from the muscadine flowers, and twenty volatiles in their corresponding hydrosols. The composition of volatiles in berry skins differed significantly from that of the vine flowers. The antioxidant potential of investigated essential oils and hydrosols was evaluated using five in vitro assays: DPPH (2,2-diphenyl-1-picrylhydrazyl) method, TEAC (Trolox equivalent antioxidant capacity), FRAP (Ferric reducing antioxidant power), CUPRAC (cupric ion reducing antioxidant capacity), and NO (nitric oxide radical scavenging assay). The essential oils from the flowers of both cultivars showed the strongest antioxidant power, whereas the hydrosols were the significantly less active. All investigated essential oils showed very weak antibacterial activities against Bacillus cereus, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. However, the essential oils from the flowers of both cultivars showed moderate antifungal activities against Candida albicans, which were stronger for the oil from "Carlos" (white muscadine cultivar). To the best of our knowledge, this is the first report on obtaining and characterizing essential oils and hydrosols from muscadine grapes. This study demonstrated the variations in aromatic compounds accumulated in flowers and mature berry skins of muscadine grapes, and evaluated their possible antioxidant and antimicrobial activities. The presented results will be the base for future research, focused on a better understanding of the molecular and regulatory mechanisms involved in aromatic compound biosynthesis and accumulation in muscadine grapes. [ABSTRACT FROM AUTHOR]

The time course of growth and betalains biosynthesis by Beta vulgaris L. (cv. Detroit Dark Red) hairy root culture were investigated. It was established that hairy root culture produced 42.2 mg/flasks betalains (26.2 mg/flasks betaxanthins and 16.0 betacyanins) between 6th and 15th days of cultivation. The uptake of sucrose, nitrate and phosphate ions were followed during the cultivation and data on the physiology of the B. vulgaris hairy root culture are presented. [Copyright &y& Elsevier]

Carotenes have been identified as the most essential carotenoids in the human body, together with lycopene, lutein, and β-cryptoxanthin due to their beneficial antioxidant activities. Because of these properties, we have designed and conducted in 2018--2021 an experiment where various tomato accessions were metabolically profiled for their antioxidant content and other fruit parameters for further usage in crop improvement programs. Same genotypes were phenotyped and profiled with DNA markers to characterize their relative genetic distance. Aggregated data from metabolic and genetic profiling were complemented by the phenotypic data to select for the most promising cross (Pl. karotina x L1116) that was performed and developed into F1 and F2 generations. After phenotyping and genetic profiling of the 104 F2 individuals obtained from that cross, 24 individuals were selected for metabolic profiling for antioxidant contents. In relation to carotenoids accumulated in fully matured fruits the highest lycopene concentration detected in our study was 587.03 µg/g DW, while β-carotene reached 440.29 µg/g DW. Our results support the use of such integrative approach to accelerate tomato breeding when improved metabolic content of the fruits is aimed. [ABSTRACT FROM AUTHOR]

Light can effectively interrogate biological systems in a reversible and physiologically compatible manner with high spatiotemporal precision. Understanding the biophysics of photo‐induced processes in bio‐systems is crucial for achieving relevant clinical applications. Employing membranes doped with the photolipid azobenzene‐phosphatidylcholine (azo‐PC), a holistic picture of light‐triggered changes in membrane kinetics, morphology, and material properties obtained from correlative studies on cell‐sized vesicles, Langmuir monolayers, supported lipid bilayers, and molecular dynamics simulations is provided. Light‐induced membrane area increases as high as ≈25% and a ten‐fold decrease in the membrane bending rigidity is observed upon trans‐to‐cis azo‐PC isomerization associated with membrane leaflet coupling and molecular curvature changes. Vesicle electrodeformation measurements and atomic force microscopy reveal that trans azo‐PC bilayers are thicker than palmitoyl‐oleoyl phosphatidylcholine (POPC) bilayers but have higher specific membrane capacitance and dielectric constant suggesting an increased ability to store electric charges across the membrane. Lastly, incubating POPC vesicles with azo‐PC solutions results in the insertion of azo‐PC in the membrane enabling them to become photoresponsive. All these results demonstrate that light can be used to finely manipulate the shape, mechanical and electric properties of photolipid‐doped minimal cell models, and liposomal drug carriers, thus, presenting a promising therapeutic alternative for the repair of cellular disorders. [ABSTRACT FROM AUTHOR]

Gardenia jasminoides Ellis is an aromatic and medicinal plant of high economic value. Much research has focused on the phytochemistry and biological activities of Gardenia fruit extracts; however, the potential of the Gardenia plant in vitro cultures used as mass production systems of valuable secondary metabolites has been understudied. This paper presents data on metabolite profiling (GC/MS and HPLC), antioxidant activities (DPPH, TEAC, FRAP, and CUPRAC), and SSR profiles of G. jasminoides plant leaves and in vitro cultures with different levels of differentiation (shoots, callus, and cell suspension). The data show strong correlations (r = 0.9777 to r = 0.9908) between antioxidant activity and the concentrations of chlorogenic acid, salicylic acid, rutin, and hesperidin. Eleven co-dominant microsatellite simple sequence repeats (SSRs) markers were used to evaluate genetic variations (average PIC = 0.738 ± 0.153). All of the investigated Gardenia in vitro cultures showed high genetic variabilities (average Na = 5.636 ± 2.157, average Ne = 3.0 ± 1.095). This is the first report on a study on metabolite profiles, antioxidant activities, and genetic variations of G. jasminoides in vitro cultures with different levels of differentiation. [ABSTRACT FROM AUTHOR]

The Amaryllidaceae alkaloid galanthamine (Gal) is a long-acting, selective, reversible, and competitive acetylcholinesterase inhibitor used for the treatment of early- to mid-stage Alzheimer's disease, poliomyelitis, and other neurological diseases. Currently, Gal is produced by extraction from plants such as daffodils ( Narcissus cultivar Carlthon), snowflake ( Leucojum aestivum), 'red-tubed lily' ( Lycoris radiata), and Ungernia victoria, and alternatively by chemical synthesis. Due to the increased demand by the generic pharmaceutical companies and the limited availability of plant sources, the biosynthesis of Gal by plant in vitro systems has been considered as an alternative approach for its sustainable production. The present article reviews the state of the art of in vitro Gal biosynthesis including growth regulators, medium components, culture conditions, elicitation, and bioreactor systems. It may be used as a starting point for further studies in this area leading to a progress in biotechnological production of this valuable alkaloid. [ABSTRACT FROM AUTHOR]

Plant cells contain a wide range of interesting secondary metabolites, which are used as natural pigments and flavoring agents in foods and cosmetics as well as phyto-pharmaceutical products. However, conventional industrial extraction from whole plants or parts of them is limited due to environmental and geographical issues. The production of secondary metabolites from in vitro cultures can be considered as alternative to classical technologies and allows a year-round cultivation in the bioreactor under optimal conditions with constant high-level quality and quantity. Compared to plant cell suspensions, differentiated plant in vitro systems offer the advantage that they are genetically stable. Moreover, the separation of the biomass from culture medium after fermentation is much easier. Nevertheless, several investigations in the literature described that differentiated plant in vitro systems are instable concerning the yield of the target metabolites, especially in submerged cultivations. Other major problems are associated with the challenges of cultivation conditions and bioreactor design as well as upscaling of the process. This article reviews bioreactor designs for cultivation of differentiated plant in vitro systems, secondary metabolite production in different bioreactor systems as well as aspects of process control, management, and modeling and gives perspectives for future cultivation methods. [ABSTRACT FROM AUTHOR]

Abstract: Calli, shoot-clumps and regenerated plants were initiated from young fruits of Pancratium maritimum L. Their genetic stability was monitored by flow cytometry before chemical studies. Apolar metabolites (alkaloids extracted at pH > 7, free fatty acids and fatty alcohols, sterols etc.) were qualitatively and quantitatively analyzed by GC–MS. The results clearly demonstrated that alkaloid synthesis in P. maritimum is closely related with tissue differentiation. The highest amounts of alkaloids and presence of homolycorine and tazettine type compounds (end products of the biosynthetic pathway of the Amaryllidaceae alkaloids) were found in highly differentiated tissues. Galanthamine accumulated in the leaves of plantlets. The amount of hordenine, a protoalkaloid, is related with the ability of tissues to synthesize alkaloids. Saturated fatty acids were found in considerably higher levels in undifferentiated callus cultures and partially differentiated shoot-clumps than in regenerated plants. Mono- and dienoic fatty acids were found at higher levels in non-photosynthesizing tissues – calli, and in vitro and intact bulbs, while α-linolenic acid (trienoic acid) was found in higher amounts in the photosynthesizing leaves of shoot-clumps and regenerated plants than in bulbs and calli. Fatty alcohols were found mainly in leaves, while sterols tended to accumulate in photosynthesizing and undifferentiated tissues. [ABSTRACT FROM AUTHOR]

Abstract: The effects of the hexapeptide angiotensin II (3–8) ANG IV, the selective A1 receptor agonist cyclohexyladenosine (CHA) and the combination of ANG IV + CHA on pentylenetetrazol (PTZ)-generalized seizures; kindling development and maintenance were studied. By using in vitro quantitative receptor autoradiography, the regulation of adenosine A1 receptor density at different time points during the kindling procedure and postkindling period was determined. ANG IV and CHA effectively reduced clonic seizures in PTZ-generalized seizure model, in PTZ-kindled mice as well as during kindling development and a week later by rechallenge with PTZ. Furthermore, coadministration of ANG IV and CHA had a strong anticonvulsant effect, both compounds acting synergistically. A significant increase of adenosine A1 receptor density was detected in somatosensory cortex, hippocampus, amygdala and geniculate nuclei early in the kindling procedure (after the 3rd injection), which persisted at least 1 month after the end of kindling procedure. In addition, a delayed up-regulation of adenosine A1 receptor binding was observed a week after kindling in the mamillary bodies and a month later in the motor cortex. The pretreatment with ANG IV caused a down-regulation of adenosine A1 receptor density to the control level in most time points and brain areas. In conclusion, PTZ kindling-induced increase of adenosine A1 receptor binding at different time points and in specific brain structures might represent an adaptive mechanism for coping with the hyperexcitability typical for this phenomenon. The antiepileptogenic effect of ANG IV could be realized partly through an adenosine-dependent mechanism. [Copyright &y& Elsevier]

Two‐phase bioreactor cultivation system was developed and applied for in sito recovery of extracellular galanthamine during the cultivation of Leucojum aestivum L. shoot culture in a modified column bioreactor system. The inclusion of an external circulation column with adsorbent resin Amberlite XAD‐4 as a second phase, on the 21st day of the beginning of cultivation resulted in 1.25 folds increase in biomass accumulation and maximal amounts of accumulated galanthamine of 6 mg/L (3.1 mg/L intracellular and 2.9 mg/L extracellular). It was demonstrated that the inclusion of a second phase at the cultivation of the L. aestivum shoot culture in a bubble column bioreactor with internal sections redirected the alkaloid metabolism to galanthamine synthesis and inhibits the synthesis of hemanthamine and lycorine type alkaloids. Our research demonstrated that the application of the two‐phase cultivation systems could be an important tool to increase the yields of valuable secondary metabolites in plant tissue culture‐based bioprocess. [ABSTRACT FROM AUTHOR]

Chemical composition and bioactivity of essential oil from Salvia tomentosa Mill. natively grown in Bulgaria were investigated. GC-MS analysis identified 60 compounds which represented 98% of the oil constituents. The prevalent constituents were monoterpenes with eight dominant compounds being identified: borneol (10.3%), β-pinene (9%), camphor (7.9%), α-pinene (6%), camphene (4%), 1.8-cineole (3.8%), α-limonene (3.5%) and β-caryophyllene (3%). The essential oil showed considerable acetylcholinesterase inhibitory activity (IC50=0.28±0.06 μg/mL), comparable with that of galanthamine. Study of antioxidant activity strongly suggested that the hydrogen atom transfer reaction was preferable over the electron transfer (ORAC=175.0±0.40 μM Trolox equivalents/g oil and FRAP=1.45±0.21 mM Trolox equivalents/g oil). The essential oil showed moderate antifungal and antibacterial activities against Candida albicans and Gram-positive bacteria, whereas it was almost inactive against the investigated Gram-negative strains. The results suggested that the essential oil of Bulgarian S. tomentosa could be considered as a prospective active ingredient for prevention of oxidative stress-related and neurodegenerative disorders in aromatherapy. Because of the high antioxidant capacity, the oil could be considered as natural supplement or antioxidant in cosmetics and food products. [ABSTRACT FROM AUTHOR]

Abstract: The callus induction from young fruits of Leucojum aestivum was performed on Murashige–Skoog nutrient medium supplemented with 4mg/L 2,4 dichlorphenoxyacetic and 2mg/L 6-benzylamynopurine. Further, by planting the obtained calluses on the same nutrient medium supplemented with 1.15mg/L α-naphtylacetic acid and 2.0mg/L 6-benzylamynopurine shoot cultures were established. The growth and galanthamine accumulation of obtained L. aestivum in vitro systems were studied. It was established that the amount of accumulated galanthamine strongly depended on the level of the differentiation. The maximum yield of biomass (17.8g/L) and the maximum amount of accumulated galanthamine (2.5mg/L) were achieved after day 35 of submerged cultivation of L. aestivum 80 shoot culture, performed under illumination. Data concerning the time courses of the utilization of the main nutrient components of the medium during cultivation of L. aestivum shoot culture are presented as well. [Copyright &y& Elsevier]