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Determining and improving drought-tolerant cultivars is a major goal of plant breeding to face climate change. The productivity of faba bean in Egypt is affected by abiotic stresses, especially drought stress. This study evaluated eight Egyptian faba bean cultivars for drought tolerance under three soil water regimes consisting of well-watered (100% field capacity), moderate drought stress (50% field capacity), and severe drought stress (25% field capacity) regimes in pots under greenhouse conditions using biochemical, physiochemical, and molecular parameters. The cultivars Nubariya 1, Nubariya 3, and Giza 716 showed the highest proline content values under 50% field capacity conditions, with 4.94, 4.39, and 4.26 mmol/g fresh weights, respectively. On the other hand, the cultivars Sakha 1, Sakha 4, Nubariya 1, and Nubariya 3 exhibited the highest proline contents (7.8, 7.53, 6.17, and 6.25, respectively) under 25% field capacity treatment. The molecular profiling was conducted using SCoT and SRAP approaches. Fresh leaves were utilized to extract the DNA, and ten primers for SRAP and six for SCoT were used in the PCR procedures. SCoT and SRAP-PCR generated 72 loci, of which, 55 were polymorphic, and 17 were monomorphic. SCoT and SRAP each had 48 and 24 total loci, respectively. The average polymorphism (%) values achieved via SCoT and SRAP were 70.93% and 80%, respectively. Based on the molecular profiles, the cluster analysis identified three clusters. The first cluster comprised Giza 716 cultivars; the second cluster included Sakha 1, Sakha 3, Sakha 4, and Akba 3300 cultivars; the third cluster comprised two cultivars Nubariya 1 and Nubariya3. According to the study’s findings, Sakha 1, Sakha 4, Nubariya 1, and Nubariya 3 are remarkable parents for developing drought-tolerant faba bean genotypes. Additionally, this study concluded that SRAP and SCoT markers recreated trustworthy banding profiles to evaluate the genetic polymorphism among faba bean cultivars, which are regarded as the cornerstone for genetic improvements in crops.

A number of phytochemicals have been identified as promising drug molecules against a variety of diseases using an in-silico approach. The current research uses this approach to identify the phyto-derived drugs from Andrographis paniculata (Burm. f.) Wall. ex Nees (AP) for the treatment of diphtheria. In the present study, 18 bioactive molecules from Andrographis paniculata (obtained from the PubChem database) were docked against the diphtheria toxin using the AutoDock vina tool. Visualization of the top four molecules with the best dockscore, namely bisandrographolide (−10.4), andrographiside (−9.5), isoandrographolide (−9.4), and neoandrographolide (−9.1), helps gain a better understanding of the molecular interactions. Further screening using molecular dynamics simulation studies led to the identification of bisandrographolide and andrographiside as hit compounds. Investigation of pharmacokinetic properties, mainly ADMET, along with Lipinski’s rule and binding affinity considerations, narrowed down the search for a potent drug to bisandrographolide, which was the only molecule to be negative for AMES toxicity. Thus, further modification of this compound followed by in vitro and in vivo studies can be used to examine itseffectiveness against diphtheria.

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PADs are a group of calcium-dependent enzymes that play key roles in inflammatory pathologies and have diverse roles in cancers. PADs cause irreversible post-translational modification of arginine to citrulline, leading to changes in protein function in different cellular compartments. PAD isozyme diversity differs throughout phylogeny in chordates, with five PAD isozymes in mammals, three in birds, and one in fish. While the roles for PADs in various human cancers are mounting (both in regards to cancer progression and epigenetic regulation), investigations into animal cancers are scarce. The current pilot-study therefore aimed at assessing PAD isozymes in a range of animal cancers across the phylogeny tree. In addition, the tissue samples were assessed for total protein deimination and histone H3 deimination (CitH3), which is strongly associated with human cancers and also indicative of gene regulatory changes and neutrophil extracellular trap formation (NETosis). Cancers were selected from a range of vertebrate species: horse, cow, reindeer, sheep, pig, dog, cat, rabbit, mink, hamster, parrot, and duck. The cancers chosen included lymphoma, kidney, lung, testicular, neuroendocrine, anaplastic, papilloma, and granulosa cell tumour. Immunohistochemical analysis revealed that CitH3 was strongly detected in all of the cancers assessed, while pan-deimination detection was overall low. Both PAD2 and PAD3 were the most predominantly expressed PADs across all of the cancers assessed, while PAD1, PAD4, and PAD6 were overall expressed at lower, albeit varying, levels. The findings from this pilot study provide novel insights into PAD-mediated roles in different cancers across a range of vertebrate species and may aid in the understanding of cancer heterogeneity and cancer evolution.

Corn tassel (CT) is a waste part of the corn plant. It is a good co-product and rich in terms of bioactive compounds and phytochemicals. This research tried to show the phenolic profile, antioxidants, anticholinergic activities, and antibacterial properties of CT ethanol extract. The phenolic content analysis of the CT was determined quantitatively by LC-MS/MS, and the antioxidant capacity was measured using ABTS, DPPH, Cu2+–Cu+, and Fe3+–Fe2+ reducing methods. The anticholinergic measurements of CT were detected by inhibition of acetylcholinesterase (AChE). The antibacterial activity was determined by MIC and disc diffusion methods. Many phenolic compounds such as vanillic acid, caffeic acid, fumaric acid, acetohydroxamic acid, butein, myricetin, resveratrol, catechin hydrate, and 4-hydroxybenzoic acid were detected in ethanol extract of CT. The obtained plant ethanol extract had a 7.04% DPPH value, while it showed ABTS activity at 9.45%. Moreover, it had a 0.10 mg/mL inhibition effect on the AChE in terms of IC50 values. The ethanol extract of the CT had an antibacterial property on the investigated bacteria at different ratios. In conclusion, this research aims to consider CT as a source of phenolic compounds and to reveal its bioactive properties and its effects on the treatment of some diseases.

Determining the appropriate parents for breeding programs is the most important decision that plant breeders must make to maximize the genetic variability and produce excellent recombinant genotypes. Several methods are used to identify genotypes with desirable phenotypic features for breeding experiments. In this study, five kalanchoe genotypes were morphologically characterized by assessing plant height, number of inflorescences, number of flowers, flower length, flower diameter and number of petals. The analysis showed the distinction of yellow kalanchoe in the plant height trait, while the orange kalanchoe was distinguished in the number of inflorescences, the number of flowers and flower length traits, whereas the violet kalanchoe possessed the largest flower diameter and the highest number of petals. The molecular profiling was performed by random amplified polymorphism DNA (RAPD), inter-simple sequence repeats (ISSR) and start codon targeted (SCoT)-polymerase chain reaction (PCR) tools. Genomic DNA was extracted from young leaves and the PCR reactions were performed using ten primers for each SCoT, ISSR and RAPD marker. Only four out of ten primers showed amplicon profiles in all PCR markers. A total of 70 bands were generated by SCoT, ISSR and RAPD-PCR with 35 polymorphic bands and 35 monomorphic bands. The total number of bands of RAPD, ISSR and SCoT was 15, 17 and 38, respectively. The polymorphism percentages achieved by RAPD, ISSR and SCoT were 60.25%, 15% and 57%, respectively. The cluster analysis based on morphological data revealed two clusters. Cluster I consisted of violet and orange kalanchoe, and cluster II comprised red, yellow and purple kalanchoe. Whereas the cluster analysis based on molecular data revealed three clusters. Cluster I included only yellow kalanchoe, cluster II comprised orange and violet kalanchoe while cluster III comprised red, and purple kalanchoe. The study concluded that orange, violet and yellow kalanchoe are distinguished parents for breeding economically valued traits in kalanchoe. Also, the study concluded that SCoT and RAPD markers reproduced reliable banding patterns to assess the genetic polymorphism among kalanchoe genotypes that consider the basis stone for genetic improvements in ornamental plants.

Stilbenes are polyphenolic allelochemicals synthesized by plants, especially grapes, peanuts, rhubarb, berries, etc., to defend themselves under stressful conditions. They are now exploited in medicine for their antioxidant, anti-proliferative and anti-inflammatory properties. Inflammation is the immune system’s response to invading bacteria, toxic chemicals or even nutrient-deprived conditions. It is characterized by the release of cytokines which can wreak havoc on healthy tissues, worsening the disease condition. Stilbenes modulate NF-κB, MAPK and JAK/STAT pathways, and reduce the transcription of inflammatory factors which result in maintenance of homeostatic conditions. Resveratrol, the most studied stilbene, lowers the Michaelis constant of SIRT1, and occupies the substrate binding pocket. Gigantol interferes with the complement system. Besides these, oxyresveratrol, pterostilbene, polydatin, viniferins, etc., are front runners as drug candidates due to their diverse effects from different functional groups that affect bioavailability and molecular interactions. However, they each have different thresholds for toxicity to various cells of the human body, and thus a careful review of their properties must be conducted. In animal models of autoinflammatory diseases, the mode of application of stilbenes is important to their absorption and curative effects, as seen with topical and microemulsion gel methods. This review covers the diversity seen among stilbenes in the plant kingdom and their mechanism of action on the different inflammatory pathways. In detail, macrophages’ contribution to inflamed conditions in the liver, the cardiac, connective and neural tissues, in the nephrons, intestine, lungs and in myriad other body cells is explored, along with detailed explanation on how stilbenes alleviate the symptoms specific to body site. A section on the bioavailability of stilbenes is included for understanding the limitations of the natural compounds as directly used drugs due to their rapid metabolism. Current delivery mechanisms include sulphonamides, or using specially designed synthetic drugs. It is hoped that further research may be fueled by this comprehensive work that makes a compelling argument for the exploitation of these compounds in medicine.

Hydroponic systems are known to provide a platform for uniform growth conditions until the reproductive stage. However, many plant species, including sunflower, show poor growth and survivability under conventional hydroponic systems due to poor nutrient availability, hypoxia and algal contamination. Thus, we tested various hydroponic systems to select a hydroponic system suitable for screening of sunflower germplasm. Sunflower accessions showed better growth and leaf gas exchange in newly-designed over conventional hydroponic systems. Selected hydroponic systems were further engaged in sunflower accession screening under heat and osmotic stress in a two-pan system (210 cm × 60 cm). Heat stress treatment was applied by growing sunflower germplasm at 42 °C and osmotic stress by adding polyethylene glycol 8000 which decreased the osmotic potential to − 0.6 MPa. There was significant variability among the sunflower accessions for their ability to survive under stress. Accessions such as C-2721 (43%), C-291 (46%) and D-14 (43%) had lower cell membrane injury percentage under osmotic stress and high seedling survivability (60‒80%) under heat stress when compared with susceptible accessions. Moreover, resistant accessions exhibited greater cuticular waxes and root length but lower transpiration losses. The newly designed hydroponic platform proved reliable for the selection of resistant sunflower accessions. Selected parental lines were validated by assessing their hybrids under field trials across two seasons under water and temperature stress during the reproductive phase (autumn). Hybrid H3 obtained by crossing drought and heat resistant parents had the highest seed yield and water use efficiency.

Microorganisms dwell in diverse plant niches as non-axenic biotic components that are beneficial as well pathogenic for the host. They improve nutrients-uptake, stress tolerance, phytohormone synthesis, and strengthening the defense system through phyllosphere, rhizosphere, and endosphere. The negative consequences of the microbial communities are largely in the form of diseases characterized by certain symptoms such as gall, cankers, rots etc. Uncultivable and unspecified nature of different phytomicrobiomes communities is a challenge in the management of plant disease, a leading cause for the loss of the plant products. Metagenomics has opened a new gateway for the exploration of microorganisms that are hitherto unknown, enables investigation of the functional aspect of microbial gene products through metatranscriptomics and metabolomics. Metagenomics offers advantages of characterizing previously unknown microorganisms from extreme environments like hot springs, glaciers, deep seas, animal gut etc. besides bioprospecting gene products such as Taq polymerase, bor encoded indolotryptoline, hydrolases, and polyketides. This review provides a detailed account of the phytomicrobiome networks and highlights the importance and limitations of metagenomics and other meta-omics approaches for the understanding of plant microbial diversity with special focus on the disease control and its management.

Hydroxylated polyphenols, also called flavonoids, are richly present in vegetables, fruits, cereals, nuts, herbs, seeds, stems, and flowers of numerous plants. They possess numerous medicinal properties such as antioxidant, anti-cancer, anti-microbial, neuroprotective, and anti-inflammation. Studies show that flavonoids activate antioxidant pathways that render an anti-inflammatory effect. They inhibit the secretions of enzymes such as lysozymes and β-glucuronidase and inhibit the secretion of arachidonic acid, which reduces inflammatory reactions. Flavonoids such as quercetin, genistein, apigenin, kaempferol, and epigallocatechin 3-gallate modulate the expression and activation of a cytokine such as interleukin-1beta (IL-1β), Tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin-8 (IL-8); regulate the gene expression of many pro-inflammatory molecules such s nuclear factor kappa-light chain enhancer of activated B cells (NF-κB), activator protein-1 (AP-1), intercellular adhesion molecule-1 (ICAM), vascular cell adhesion molecule-1 (VCAM), and E-selectins; and also inhibits inducible nitric oxide (NO) synthase, cyclooxygenase-2, and lipoxygenase, which are pro-inflammatory enzymes. Understanding the anti-inflammatory action of flavonoids provides better treatment options, including coronavirus disease 2019 (COVID-19)-induced inflammation, inflammatory bowel disease, obstructive pulmonary disorder, arthritis, Alzheimer’s disease, cardiovascular disease, atherosclerosis, and cancer. This review highlights the sources, biochemical activities, and role of flavonoids in enhancing human health.

Common fig (Ficus carica L.), an economically-important fruit species widely cultivated in the Mediterranean Basin, is gaining increasing interest in the tropical and subtropical regions. A vast genetic diversity of common fig is available; however, systematic studies of cultivar performance in a particular new environment are scarce. The objective of this study was to evaluate the performance of 17 common fig cultivars grown under the Malaysian tropical climate. Evaluation was based on 15 parameters relevant to fruit yield, fruit quality and pest susceptibility. The results showed variable performance among the cultivars tested; 10 cultivars performed well for orchard establishment. Cultivar ‘A134’ produced the highest fruit yield per plant (3410 g) compared to the average yield of the cultivars studied (2500 g). Cultivars ‘Fen Chan Huang’ and ‘Wuhan’ produced the highest number of fruits per plant, 119 and 82 fruits, respectively. Mealybug infestation and ostiole-end splitting are major limitations to fig cultivation in Malaysia. Cultivar ‘Longue d’Aout’ was the most susceptible to mealybug infestation; whereas cultivar ‘B110’ was the most resistant. Above 60% of the fruits of cvs. ‘Brunswick’ and ‘Qing Pi’ had split ostiole; however, this was less than 20% in cvs. ‘Longue d’Aout’ and ‘Alma’. This study facilitates successful farming of figs in Malaysia and provides insights for future development programs.

A common modern cultivation practice is bagging the fruit bunch if date palm (Phoenix dactylifera L.) which may influence fruit maturity and nutraceutical quality. Exposure of fruits to photooxidative stress induces changes in the endogenous concentrations of plant hormones and other metabolites, which may cause accelerated fruit maturity. This study was conducted to examine the effect of exposure to direct and indirect sunlight on date palm fruit development. The indirect sunlight treatment was simulated by fruit bunch bagging, a common practice in modern date production. The exposure of date palm fruits to direct sunlight-induced photooxidative stress causing an increased concentration of ascorbic acid and decreased content of chlorophyll, anthocyanins, carotenoids, and phenols compared to the fruit bagging treatment. Direct sunlight also reduced the concentration of phytohormones, including indoleacetic acid, gibberellin, and zeatin, but increased abscisic acid accumulation. The directly-exposed fruits reached a partially-mature stage (Rutab) in August, whereas the bagged fruits remained at the immature stage (Khalal). This study is the first to describe the biochemical basis of the observed improvement of date palm fruit development in response to reduced light intensity. Besides, it provides insights into controlling date palm fruit maturity and subsequently prolonging the shelf life dates on the tree; thus, extending the marketing period for the benefit of the farmers.

The threat of desertification is considered a global concern that occurs in many environments in different parts of the world, where extensive lands are transformed gradually into desert or semi-desert areas, and this causes economic and health issues. Iraq and many other parts of the Middle East are facing desertification threats in the last twenty years. Despite the significance of this issue, relevant reviews are scarce. The removal of vegetation cover, overgrazing, deforestation in times of war, poor irrigation practices and water scarcity are some of the main causes of desertification in Iraq. Fighting desertification requires cooperative efforts including the utilization of innovative practices, biotechnological approaches, restoration of oases, continuous reforestation, and rehabilitation of agricultural lands. The objective of this review article is to discuss the causes of desertification and land degradation in Iraq, highlighting the main natural and human factors involved, and the consequent impact on the national security, economy, society, and health. In addition, it suggests recommendations for policies and actions that can be integrated to mitigate this problem.

Essential oils are hydrophobic liquids produced as secondary metabolites by specialized secretory tissues in the leaves, seeds, flowers, bark and wood of the plant, and they play an important ecological role in plants. Essential oils have been used in various traditional healing systems due to their pharmaceutical properties, and are reported to be a suitable replacement for chemical and synthetic drugs that come with adverse side effects. Thus, currently, various plant sources for essential oil production have been explored. Coriander essential oil, obtained from the leaf and seed oil of Coriandrum sativum, has been reported to have various biological activities. Apart from its application in food preservation, the oil has many pharmacological properties, including allelopathic properties. The present review discusses the phytochemical composition of the seed and leaf oil of coriander and the variation of the essential oil across various germplasms, accessions, at different growth stages and across various regions. Furthermore, the study explores various extraction and quantification methods for coriander essential oils. The study also provides detailed information on various pharmacological properties of essential oils, such as antimicrobial, anthelmintic, insecticidal, allelopathic, antioxidant, antidiabetic, anticonvulsive, antidepressant, and hepatoprotective properties, as well as playing a major role in maintaining good digestive health. Coriander essential oil is one of the most promising alternatives in the food and pharmaceutical industries.

Neonatologists usually wait 48 hours for blood culture results before deciding to discontinue antibiotics. The objective of the study was to analyze time to positive blood culture in rule out sepsis and estimate the minimum duration of antibiotics.Retrospective analysis of blood culture at the Neonatal Intensive Care Unit, McMaster Children's Hospital (January 2004 to December 2013) using BacT/Alert® 3D microbial system was conducted. We calculated average time taken for blood culture samples to emit a positive signal and compared it between Gram-positive and Gram-negative organisms. Kaplan-Meier curves for time to detect positive culture were generated. A Cox proportional hazard regression model with the outcome variable "time to detect positive blood culture" and predictor variables "early-onset sepsis (EOS) versus late-onset sepsis (LOS)", "Gram-positive versus Gram-negative" and "definite versus possible pathogen versus contaminant" was generated.Of 7,480 blood cultures performed in 9,254 neonates, 885 samples grew microorganisms. 845 culture reports from 627 neonates were analyzed. Definite or opportunistic pathogens caused 815 (96%) infections (54 EOS and 791 LOS) and the rest were contaminants. Gram-negative organisms grew significantly faster than Gram-positive (P0.001). Cultures from EOS were positive significantly earlier than LOS (P = 0.032). Gram-negative status was an independent predictor of early detection of a positive culture (hazard ratio 3.5 [95% CI, 2.7-4.5] P0.001).The probability of positive blood culture beyond 24 hours for a Gram-negative organism is small. Empiric antimicrobial treatment can be reduced after 24 hours to target only Gram-positive organisms in LOS and can be stopped in EOS unless clinical or laboratory parameters strongly suggest sepsis.

Global technological advancements drive daily energy consumption, generating additional carbon-induced climate challenges. Modifying process parameters, optimizing design, and employing high-performance working fluids are among the techniques offered by researchers for improving the thermal efficiency of heating and cooling systems. This study investigates the heat transfer enhancement of hybrid “Al2O3-Cu/water” nanofluids flowing in a two-dimensional channel with semicircle ribs. The novelty of this research is in employing semicircle ribs combined with hybrid nanofluids in turbulent flow regimes. A computer modeling approach using a finite volume approach with k-ω shear stress transport turbulence model was used in these simulations. Six cases with varying rib step heights and pitch gaps, with Re numbers ranging from 10,000 to 25,000, were explored for various volume concentrations of hybrid nanofluids Al2O3-Cu/water (0.33%, 0.75%, 1%, and 2%). The simulation results showed that the presence of ribs enhanced the heat transfer in the passage. The Nusselt number increased when the solid volume fraction of “Al2O3-Cu/water” hybrid nanofluids and the Re number increased. The Nu number reached its maximum value at a 2 percent solid volume fraction for a Reynolds number of 25,000. The local pressure coefficient also improved as the Re number and volume concentration of “Al2O3-Cu/water” hybrid nanofluids increased. The creation of recirculation zones after and before each rib was observed in the velocity and temperature contours. A higher number of ribs was also shown to result in a larger number of recirculation zones, increasing the thermal performance.

Environmental pollution is one of the most pressing challenges in today’s world. The main cause of this pollution is fuel emissions from automobiles and other sources. As industrialization progresses, we will be unable to compromise on the use of energy to power heavy machines and will be forced to seek out the best options. As a consequence, utilizing green fuel, such as biodiesel derived from natural sources, is a realistic option. Jatropha curcas L. (Euphorbiaceae) is recognized as the greatest feedstock for biodiesel production throughout the world, and it has gained a huge market value in the recent years. Conventional cultivation alone will not be sufficient to meet the global need for the plant’s biomass for the production of biodiesel. Adoption of plant tissue culture techniques that improve the biomass availability is an immediate need. The present review provides detailed information regarding in-vitro plant propagation (direct and indirect organogenesis), somatic embryogenesis, and acclimatization protocols of plantlets for stabilized production of biomass. The review also focuses on biotechnological approaches such as gene transformation studies, production of haploids, and double haploids for developing elite germplasm for high biomass and improved traits for the production of biodiesel.

Artemisinin is an anti-malarial sesquiterpene lactone derived from Artemisia annua L. (Asteraceae family). One of the most widely used modes of treatment for malaria is an artemisinin-based combination therapy. Artemisinin and its associated compounds have a variety of pharmacological qualities that have helped achieve economic prominence in recent years. So far, research on the biosynthesis of this bioactive metabolite has revealed that it is produced in glandular trichomes and that the genes responsible for its production must be overexpressed in order to meet demand. Using biotechnological applications such as tissue culture, genetic engineering, and bioreactor-based approaches would aid in the upregulation of artemisinin yield, which is needed for the future. The current review focuses on the tissue culture aspects of propagation of A. annua and production of artemisinin from A. annua L. cell and organ cultures. The review also focuses on elicitation strategies in cell and organ cultures, as well as artemisinin biosynthesis and metabolic engineering of biosynthetic genes in Artemisia and plant model systems.

Aedes mosquitoes are implicated in the transmission of several viruses, including Dengue virus (DENV) to millions of people worldwide. The global expansion of Aedes mosquitos'habitats creates a desperate need for control mechanisms with minimum negative effects. Deciphering the molecular interactions between DENV and its vector is a promising field to develop such efficient control strategies. As soon as the viremic blood is ingested by the mosquito, DENV is encountered by different innate immunity responses. During the past three decades, different pathways of innate immunity have been identified in Aedes spp. Recognition of viral molecular patterns, including viral RNA, and vector attempts to resist DENV infection are the most important defense mechanisms. Crosstalk between innate immune pathways and redundancy of anti-DENV responses become more evident as research progresses. The viral evasion and repression of vector immune response are increasingly being discovered. Such viral strategies are potential targets to be disrupted in order to limit DENV infection and spread. Vector-related non-immune factors such as gut microbiota can also be tapped for efficient control of DENV infection in Aedes mosquito's populations without affecting their fitness. Current trends in controlling DENV in its vector are exploring the potentials of using genetically engineered mosquitoes via RNA-based systems to degrade DENV genome once released into the midgut cells cytoplasm at the early phase of the infection.

Plants that synthesize bioactive compounds that have high antioxidant value and elicitation offer a reliable in vitro technique to produce important nutraceutical compounds. The objective of this study is to promote the biosynthesis of these phenolic compounds on a large scale using elicitors in date palm cell suspension culture. Elicitors such as pectin, yeast extract (YE), salicylic acid (SA), cadmium chloride (CdCl2), and silver nitrate (AgNO3) at 50, 100, and 200 mg/L concentrations are used. The effects of elicitors on cell culture were determined in terms of biomass [packed cell volume (PCV), fresh and dry weight], antioxidant activity, and phenolic compounds (catechin, caffeic acid, kaempferol, apigenin) were determined using high-performance liquid chromatography (HPLC). Results revealed that enhanced PCV (12.3%), total phenolic content [317.9 &plusmn, 28.7 mg gallic acid equivalents (GAE)/100 g of dry weight (DW)], and radical scavenging activity (86.0 &plusmn, 4.5%) were obtained in the 50 mg/L SA treated cell culture of Murashige and Skoog (MS) medium. The accumulation of optimum catechin (26.6 &plusmn, 1.3 &micro, g/g DW), caffeic acid (31.4 &plusmn, 3.8 &micro, g/g DW), and kaempferol (13.6 &plusmn, 1.6 &micro, g/g DW) was found in the 50 mg/L SA-treated culture when compared to the control. These outcomes could be of great importance in the nutraceutical and agronomic industries.

The novel strain of coronavirus that appeared in 2019, SARS-CoV-2, is the causative agent of severe respiratory disease, COVID-19, and the ongoing pandemic. As for SARS-CoV that caused the SARS 2003 epidemic, the receptor on host cells that promotes uptake, through attachment of the spike (S) protein of the virus, is angiotensin-converting enzyme 2 (ACE2). In a recent article published by Batlle et al. (Clin. Sci. (Lond.) (2020) 134, 543–545) it was suggested that soluble recombinant ACE2 could be used as a novel biological therapeutic to intercept the virus, limiting the progression of infection and reducing lung injury. Another way, discussed here, to capture SARS-CoV-2, as an adjunct or alternative, would be to use ACE2+-small extracellular vesicles (sEVs). A competitive inhibition therapy could therefore be developed, using sEVs from engineered mesenchymal stromal/stem cells (MSCs), overexpressing ACE2.

A rapid economically viable micropropagation protocol has been developed in the present work for Morus indica L. (K-2 cultivar) utilizing the readily available nodal explants. Explants were established on different plant growth regulators (PGRs) either individually or in combinations. MS medium containing 1 mg L−1 Kinetin (Kin) showed the best shoot multiplication with 4.8 ± 0.23 cm average shoot length and 6.5 ± 0.03 number of internodes. Regenerated shoots were elongated in MS medium supplemented with 1.5 mg L−1 gibberellic acid (GA3). Elongated shoots cultured in full-strength MS medium supplemented with 1 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D) for one week and then cultured in half-strength MS proved to be more effective in rooting compared to other PGRs in significantly shorter duration. Micropropagated plants transferred to soil fortified with the quarter-strength of MS salts along with humidity regulation process showed 89% survival frequency. In vitro flowering in the regenerated shoots was also observed in the MS medium supplemented with (1.5 mg L−1) Kin and carbon source replaced by commercial sugar cubes. This method can be effectively used for in vitro culture of M. indica in commercial scale owing to its enhanced quality and reduced time frame.

There is an urgent need to develop new combination therapies beyond existing surgery, radio- and chemo-therapy, perhaps initially combining chemotherapy with the targeting specificities of immunotherapy. For this, strategies to limit inflammation and immuno-suppression and evasion in the tumour microenvironment are also needed. To devise effective new immunotherapies we must first understand tumour immunology, including the roles of T cells, macrophages, myeloid suppressor cells and of exosomes and microvesicles (EMVs) in promoting angiogenesis, tumour growth, drug resistance and metastasis. One promising cancer immunotherapy discussed uses cationic liposomes carrying tumour RNA (RNA-lipoplexes) to provoke a strong anti-viral-like (cytotoxic CD8+ ) anti-tumour immune response. Mesenchymal stem cell-derived EMVs, with their capacity to migrate towards inflammatory areas including solid tumours, have also been used. As tumour EMVs clearly exacerbate the tumour microenvironment, another therapy option could involve EMV removal. Affinity-based methods to deplete EMVs, including an immuno-depletion, antibody-based affinity substrate, are therefore considered. Finally EMV and exosome-mimetic nanovesicles (NVs) delivery of siRNA or chemotherapeutic drugs that target tumours using peptide ligands for cognate receptors on the tumour cells are discussed. We also touch upon the reversal of drug efflux in EMVs from cancer cells which can sensitize cells to chemotherapy. The use of immunotherapy in combination with the advent of EMVs provides potent therapies to various cancers. This article is protected by copyright. All rights reserved.

The formation of new proteins under the influence of harsh environmental conditions is a plant adaptation reaction. Two-year-old date palm tissue culture-derived plants from ‘Barhee’ grown in the field were subjected to salt stress (70 g l-1 NaCl) and dehydration-induced by applying 70 g l-1 polyethylene glycol or without irrigation and withholding irrigation (0 g l-1) for one month. The soluble carbohydrate content increased in response to salinity and polyethylene glycol treatment in leaves compared to the control and drought treatment without irrigation. Proline increased in all treatments. Malondialdehyde and hydrogen peroxide increased under salinity. Salinity treatment increased the activity of ascorbate peroxidase and catalase enzyme. Salinity and polyethylene glycol treatments increased abscisic acid, whereas the indoleacetic acid level decreased. The protein pattern of roots and leaves in one-dimensional polyacrylamide gel electrophoresis showed that the stress conditions led to new protein bands' appearance and other proteins' disappearance. A comparison of protein patterns between the control and stress treatments revealed that the relative intensity of proteins in roots and leaves were more associated with salinity treatment than the drought. The results may be clearing important the molecular mechanism of tolerance under the influence of extreme environmental stress.

Exosomes and microvesicles (EMV) are lipid bilayer-enclosed structures, released by cells and involved in intercellular communication through transfer of proteins and genetic material. EMV release is also associated with various pathologies, including cancer, where increased EMV release is amongst other associated with chemo-resistance and active transfer of pro-oncogenic factors. Recent studies show that EMV-inhibiting agents can sensitize cancer cells to chemotherapeutic agents and reduce cancer growth in vivo. Cannabidiol (CBD), a phytocannabinoid derived from Cannabis sativa, has anti-inflammatory and anti-oxidant properties, and displays anti-proliferative activity. Here we report a novel role for CBD as a potent inhibitor of EMV release from three cancer cell lines: prostate cancer (PC3), hepatocellular carcinoma (HEPG2) and breast adenocarcinoma (MDA-MB-231). CBD significantly reduced exosome release in all three cancer cell lines, and also significantly, albeit more variably, inhibited microvesicle release. The EMV modulating effects of CBD were found to be dose dependent (1 and 5 μM) and cancer cell type specific. Moreover, we provide evidence that this may be associated with changes in mitochondrial function, including modulation of STAT3 and prohibitin expression, and that CBD can be used to sensitize cancer cells to chemotherapy. We suggest that the known anti-cancer effects of CBD may partly be due to the regulatory effects on EMV biogenesis, and thus CBD poses as a novel and safe modulator of EMV-mediated pathological events.

Hypoglycemia may lead to neurological impairment; therefore, high-risk newborns are screened postnatally. However, hypoglycemia monitoring protocols often do not include cord blood acidosis as a risk factor. The study aimed to find an association between asymptomatic cord blood acidosis and hypoglycemia. All healthy term infants born at McMaster Children Hospital, Hamilton, Canada, between October 2013 and September 2014, who had umbilical cord blood pH

Exosomes, membrane vesicles of 40–100 nm in diameter, are derived from endosomes in various cells. The bioactive molecules specifically packed into exosomes can be horizontally transferred into recipient cells changing their biological properties, by which tumour cells continuously modify their surrounding microenvironment and distant target cells favouring cancer metastasis. It has been suspected for a long time that exosomes participate in the whole process of tumour metastasis. Although there is much unknown and many controversies in the role of cancer exosome, the major contribution of tumour-associated exosomes to different steps of cancer metastasis are demonstrated in this review. Mainly because these exosomes are easily accessible and capable of representing their parental cells, exosomes draw much attention as a promising biomarker for tumour screening, diagnosis and prognosis. Currently, researchers have found numerous biomarkers in exosomes with great potential to be utilized in personalized medicine. In this article, we summarize the roles of biomarkers, which are validated by clinical samples. Even though many conundrums remain, such as exosome extraction, large multicentre validation of biomarkers and data interpretation, exosomes are certain to be used in clinical practice in the near future as the field rapidly expands. Keywords: extracellular vesicle; metastasis; malignant neoplasms; microenvironment; personalized diagnostics; cancer biomarkers (Published: 22 June 2015) Citation: Journal of Extracellular Vesicles 2015, 4: 27522 - http://dx.doi.org/10.3402/jev.v4.27522

MVs are released in response to several stress agents, in an attempt to prevent continued cellular damage. After an initial stimulus of prostate cancer cells with sublytic C5b-9 and activation of MV release through PKC, cells take at least 20 min to fully recover their ability to microvesiculate. This release of MVs through activation of sublytic C5b-9 was inhibited by the PKC inhibitor bisindoylmaleimide I but not the Rho kinase inhibitor, Y27632. After stimulus there is a rise of 79 nMs(-1) over 11 s, reaching a peak [Ca(2+)]i of 920 nM. The concentration of cytosolic calcium then falls steadily at 2.4 nMs(-1) over 109 s reaching baseline levels (50-100 nM) within 10-15 min. In PC3 cells the rate of release of MVs from stimulated cells also reaches a minimum within 10-15 min. Using fura-2 AM-loaded cells, upon stimulation, cells were found to release MVs with a concentration of intravesicular calcium estimated at ∼ 430 nM.

Using a Quartz Crystal Microbalance with dissipation monitoring, QCM-D (label-free system) measuring changes in resonant frequency (Δf) that equate to mass deposited on a sensor, we showed the attachment, over a 60 min period, of a monolayer of PC3 cells to the gold electrodes of the quartz crystal sensor, which had been rendered hydrophilic. That MVs were released upon BzATP stimulation of cells was confirmed by NTA analysis (average 250 nm diameter), flow cytometry, showing high phosphatidylserine exposition and by fluorescent (Annexin V Alexa Fluor® 488-positive) and electron microscopy. Over a period of 1000s (16.7 min) during which early apoptosis increased from 4% plateauing at 10% and late apoptosis rose to 2%, the Δf increased 20 Hz, thereupon remaining constant for the last 1000s of the experiment. Using the Sauerbrey equation, the loss in mass, which corresponded to the release of 2.36 × 106 MVs, was calculated to be 23 ng. We therefore estimated the mass of an MV to be 0.24 pg. With the deposition on the QCM-D of 3.5 × 107 MVs over 200s, the decrease in Δf (Hz) gave an estimate of 0.235 pg per MV.

A cost-effective fabrication methodology for the mass production of conical microfunnels for drug delivery applications is presented. The fabrication methodology originates from a technique previously used for fabricating suspended or hollow microstructures. The process comprises six stages: in stage 1, a UV-opaque resin substrate is formed by mixing photocurable SU-8 with a UV-opaque impurity; in stage 2, an embossing stamp with an array of microrods is fabricated; in stage 3, the upper surface of the UV-opaque resin substrate (from the first stage) is mechanically deformed by stamping with the embossing stamp from stage 2; in stage 4, the regions in which microfunnels are photolithographically defined are selectively exposed to UV light; in stage 5, the wafer from stage 4 is subjected to heat and then developed in 1-methoxy-2-propanol acetate; finally, a hole on the tip of each microfunnel is formed by dry etching with oxygen plasma. The sharpness of the funnel tips can be flexibly adjusted by changing the stroke speed of the stamping in stage 3. The overall process can be carried out photolithographically with conventional photolithographic equipment. Using the proposed fabrication methodology, a matrix of polymeric microfunnels tapering from the base measuring 300 μm to the tip measuring 50 μm over a funnel height of 100 μm has been fabricated. Each fabricated microfunnel features a wide base and a sharp tip so that it has sufficient mechanical strength and sharpness to pierce the human skin. The conical geometry used in this study had a volume in excess of 24.4 nl.

A reverse phase HPLC method is developed for the simultaneous analysis of Trifluoper- azine (TFP) and prochlorperazine (PCP) in pharmaceutical preparations. HPLC was carried out on a C18 column using acetonitrile as a mobile phase at 1 mL min � 1 flow rate and the effluent was monitored at 250 nm. Chlorperazine hydrochloride (CPZ) was used as an internal standard. The retention time of the drugs was 10.879 and 13.708 min, respectively. This method produced a linear response in the concentration range between 5-200 l gm l � 1 of trifluoperazine and 10-500 l gm l � 1 of prochlorperazine. In this study, a HPLC method was successfully applied for the quantitative assay of trifluoperazine and prochlorperazine in tablets and ampule of commercial preparations which is simple, rapid and does not require any separation step for each drug.

Microvesicles are shed constitutively, or upon activation, from both normal and malignant cells. The process is dependent on an increase in cytosolic Ca2+, which activates different enzymes, resulting in depolymerization of the actin cytoskeleton and release of the vesicles. Drug resistance can be defined as the ability of cancer cells to survive exposure to a wide range of anti-cancer drugs, and anti-tumour chemotherapeutic treatments are often impaired by innate or acquired MDR (multidrug resistance). Microvesicles released upon chemotherapeutic agents prevent the drugs from reaching their targets and also mediate intercellular transport of MDR proteins.

The release of extracellular vesicles, whether MVs (microvesicles) or exosomes, from host cells or intracellular pathogens is likely to play a significant role in the infection process. Host MVs may fuse with pathogen surfaces to deliver host complement regulatory proteins. They may also deliver cytokines that enhance invasion. Decoy functions are also possible. Whereas host MVs may direct pathogens away from their target cells, pathogen MVs may in turn redirect complement membrane-attack complexes away from their target pathogen. An understanding of the mechanisms of this interplay, bringing about both immune evasion and enhanced invasion, will help to direct future research with a view to rendering pathogens more susceptible to immune attack or in improving drug efficacy. It should also be possible to use MVs or exosomes isolated directly from the pathogens, or from the cells infected with pathogens, to provide alternative vaccination strategies.

The innate immune system is the first mechanism of vertebrate defense against pathogen infection. In this study, we present evidence for a novel immune evasion mechanism of Trypanosoma cruzi, mediated by host cell plasma membrane-derived vesicles. We found that T. cruzi metacyclic trypomastigotes induced microvesicle release from blood cells early in infection. Upon their release, microvesicles formed a complex on the T. cruzi surface with the complement C3 convertase, leading to its stabilization and inhibition, and ultimately resulting in increased parasite survival. Furthermore, we found that TGF-β–bearing microvesicles released from monocytes and lymphocytes promoted rapid cell invasion by T. cruzi, which also contributed to parasites escaping the complement attack. In addition, in vivo infection with T. cruzi showed a rapid increase of microvesicle levels in mouse plasma, and infection with exogenous microvesicles resulted in increased T. cruzi parasitemia. Altogether, these data support a role for microvesicles contributing to T. cruzi evasion of innate immunity.

Microvesicles (or MVs) are plasma membrane-derived vesicles released from most eukaryotic cells constitutively during early apoptosis or at higher levels after chemical or physical stress conditions. This review looks at some of the functions of MVs in terms of intercellular communication and ensuant signal transduction, including the transport of proteins (unconventional protein export) as well as of mRNA and microRNA. MVs also have roles in membrane repair, the removal of misfolded proteins, and in the control of apoptosis. We also discuss the role MVs have been shown to have in invasive growth and metastasis as well as in hypoxia in tumours and cerebral ischaemia. The association of MVs in infectious and autoimmune disease is also summarised together with their possible use as therapeutic agents.

Complex organic additives are known to improve growth and differentiation of in vitro plant cultures. The present investigation was conducted to determine the effect of various concentrations of yeast extract (YE) and casein hydrolysate (CH) on callus growth and somatic embryogenesis in date palm cultivar Nabout Saif. Callus induced from shoot tip explants was grown on callus multiplication medium supplemented with either YE or CH at 0.0, 0.1, 0.25, 0.5, and 1 g l−1. To induce somatic embryogenesis, callus was transferred to a hormone-free medium containing the corresponding concentration of additives. The results have shown that callus weight and the number of somatic embryos were directly proportional to increases in the concentration of organic additives tested. Callus growth was best achieved when 1 g l−1 of either YE or CH was added to the culture medium. At this concentration of YE, callus growth was double that of the control medium. On CH-containing media growth was 2.3 times that of the control. This indicates that CH is more effective in enhancing callus growth. However, YE was more effective in enhancing somatic embryogenesis. The data show that the best somatic embryo formation was obtained on either 1 g l−1 YE or 0.5 g l−1 CH which produced 45 and 30 embryos per culture, respectively, as compared to 20 embryos produced in the control treatment. Resultant somatic embryos successfully rooted and regenerated plantlets which exhibited normal growth in the greenhouse. Enhanced plant regeneration, an essential criterion for commercial micropropagation, was achieved.

The methods of Plasma Membrane-derived Vesicle (PMV) isolation and quantification vary considerably in the literature and a new standard needs to be defined. This study describes a novel filtration method to isolate PMVs in plasma, which avoids high speed centrifugation, and to quantify them using a Becton Dickinson(BD) FACS Calibur (TM) flow cytometer, as annexin V-positive vesicles, larger than 0.2 mu m in diameter. Essentially microvesicles (which comprise a mixture of PMVs and exosomes) from citrate plasma were sonicated to break up clumped exosomes, and filtered using Millipore 0.1 mu m pore size Hydrophilic Durapore membranes in Swinnex 13 mm filter holders. Phosphatidylserine-positive PMVs detected with annexin V-PE were quantified using combined labelling and gating strategies in conjunction with Polysciences Polybead Microspheres (0.2 mu m) and BDTrucount tubes. The PMV absolute count was calculated on the analysis template using the Trucount tube lot number information and expressed in PMV count/ml. Having estimated a normal reference range (0.51 x 10(5)-2.82 x 10(5) PMVs/ml) from a small sample of human donors, using the developed method, the effect of certain variables was investigated. Variations such as freezing of samples and gender status did not significantly alter the PMV absolute count, and with age plasma PMV levels were only marginally reduced. Smokers appeared to have reduced PMV levels. Nicotine, as for calpeptin was shown to dose-dependently (from 10 up to 50 mu m) reduce levels of early apoptosis in THP-1 monocytes and to decrease the level of PMV release. Fasting individuals had 2-3 fold higher PMV absolute counts compared to non-fasting subjects.

Plasma membrane-derived vesicles (PMVs) or microparticles are vesicles (0.1-1mum in diameter) released from the plasma membrane of all blood cell types under a variety of biochemical and pathological conditions. PMVs contain cytoskeletal elements and some surface markers from the parent cell but lack a nucleus and are unable to synthesise macromolecules. They are also defined on the basis that in most cases PMVs express varying amounts of the cytosolic leaflet lipid phosphatidylserine, which is externalised during activation on their surface. This marks the PMV as a biologically distinct entity from that of its parent cell, despite containing surface markers from the original cell, and also explains its role in events such as phagocytosis and thrombosis. There is currently a large amount of variation between investigators with regard to the pre-analytical steps employed in isolating red cell PMVs or RPMVs (which are slightly smaller than most PMVs), with key differences being centrifugation and sample storage conditions, which often leads to result variability. Unfortunately, standardization of preparation and detection methods has not yet been achieved. This review highlights and critically discusses the variables contributing to differences in results obtained by investigators, bringing to light numerous studies of which RPMVs have been analysed but have not yet been the subject of a review.

Fluorine makes the difference: FIBX (see structure), the tetrafluoro derivative of the hypervalent iodine reagent, is more soluble and has higher reactivity than its nonfluorinated counterpart. An efficient synthesis of FIBX and initial reactions are presented. Some of these reactions can be conducted in standard organic solvents. Owing to the increased reactivity, new transformations and catalytic reactions may be possible.

Complement C2 receptor inhibitor trispanning (CRIT) inhibits the classical pathway (CP) C3 convertase formation by competing with C4b for the binding of C2. The C-terminal 11-amino-acid of the first CRIT-extracellular domain (CRIT-H17) has a strong homology with a sequence in the C4beta chain, which is responsible for the binding of C2. Since the CP and alternative pathway (AP) C3 convertases have many functional and structural similarities, we further investigated the effects of CRIT-H17 on the AP. The factor D-mediated cleavage of factor B (FB) was blocked by CRIT-H17. By ELISA and immunoblot, CRIT-H17 was shown to bind FB. CRIT-H17 had no decay activity on the C3bBb complex as compared to decay-accelerating factor. Binding of CRIT-H17 to FB did not interfere with the assembly of C3bB complex. In a haemolytic assay using C2-deficient serum, CRIT-H17 interfered with AP complement activation.

Horizontal gene transfer (HGT), in relation to genetic transfer between hosts and parasites, is a little described mechanism. Since the complement inhibitor CRIT was first discovered in the human Schistosoma parasite (the causative agent of Bilharzia) and in Trypanosoma cruzi (a parasite causing Chagas' disease), it has been found to be distributed amongst various species, ranging from the early teleost cod to rats and humans. In terms of evolutionary distance, as measured in a phylogenetic analysis of these CRIT genes at nucleotide level, the parasitic species are as removed from their human host as is the rat sequence, suggesting HGT. The hypotheses that CRIT in humans and schistosomes is orthologous and that the presence of CRIT in schistosomes occurs as a result of host-to-parasite HGT are presented in the light of empirical data and the growing body of data on mobile genetic elements in human and schistosome genomes. In summary, these data indicate phylogenetic proximity between Schistosoma and human CRIT, identity of function, high nucleotide/amino acid identity and secondary protein structure, as well as identical genomic organization.

Complement C2 receptor inhibitor trispanning (CRIT) is a receptor for the second component of complement and is found in various tissues and hemopoietic cells. On binding to CRIT, C2 cannot be activated to potentially form a variant-C3 convertase as it is rendered non-cleavable by C1s. CRIT thus limits the amount of C3 convertase formed on the cell surface. In this study we have shown, using flow cytometry and immunofluorescence microscopy, that human CRIT undergoes endocytosis from the plasma membrane. The endocytosis, possibly ligand mediated, occurs via clathrin-coated pits as it can be inhibited by prior incubation of cells in hypertonic medium or with chlorpromazine, at 37 degrees C. However, inhibition of endocytosis was not possible after treatment with nystatin, or filipin, inhibitors of caveolae/raft-dependent endocytosis. In the presence of C2 alone, CRIT associates with the adapter protein, beta-arrestin-2, and whether in association with C2 or not, then appears in the perinuclear region, but does not appear to be translocated into the nucleus. Apart from the C3aR and C5aR that internalize the anaphylatoxic peptides, this is the first report of the internalization via the clathrin pathway of a receptor for a complement serum protein.

This study was conducted to examine the response of date palm (Phoenix dactylifera L., cvs. Barhee and Hillali) calli to water stress. Callus derived from shoot tip explants was inoculated in liquid Murashige and Skoog medium containing 10 mg dm−3α-naphthaleneacetic acid, 1.5 mg dm−3 2-isopentenyladenine, and 0 to 30 % (m/v) polyethylene glycol (PEG 8000) to examine the effect of water stress. After 2 weeks, callus growth, water content, and proline accumulation were assessed. Increasing water stress caused a progressive reduction in growth as expressed in callus fresh mass, relative growth rate, and index of tolerance. Both genotypes tested followed this general trend, however, cv. Barhee was more tolerant to drought stress than cv. Hillali. Increasing PEG concentration was also associated with a progressive reduction in water content and increased content of endogenous free proline.