MicroRNAs modulating nutrient homeostasis: a sustainable approach for developing biofortified crops.

During their lifespan, sessile plants have to cope with bioavailability of the suboptimal nutrient concentration and have to constantly sense/evolve the connecting web of signal cascades for efficient nutrient uptake, storage, and translocation for proper growth and metabolism. However, environmental fluctuations and escalating anthropogenic activities are making it a formidable challenge for plants. This is adding to (micro)nutrient-deficient crops and nutritional insecurity. Biofortification is emerging as a sustainable and efficacious approach which can be utilized to combat the micronutrient malnutrition. A biofortified crop has an enriched level of desired nutrients developed using conventional breeding, agronomic practices, or advanced biotechnological tools. Nutrient homeostasis gets hampered under nutrient stress, which involves disturbance in short-distance and long-distance cell-cell/cell-organ communications involving multiple cellular and molecular components. Advanced sequencing platforms coupled with bioinformatics pipelines and databases have suggested the potential roles of tiny signaling molecules and post-transcriptional regulators, the microRNAs (miRNAs) in key plant phenomena including nutrient homeostasis. miRNAs are seen as emerging targets for biotechnology-based biofortification programs. Thus, understanding the mechanistic insights and regulatory role of miRNAs could open new windows for exploring them in developing nutrient-efficient biofortified crops. This review discusses significance and roles of miRNAs in plant nutrition and nutrient homeostasis and how they play key roles in plant responses to nutrient imbalances/deficiencies/toxicities covering major nutrients-nitrogen (N), phosphorus (P), sulfur (S), magnesium (Mg), iron (Fe), and zinc (Zn). A perspective view has been given on developing miRNA-engineered biofortified crops with recent success stories. Current challenges and future strategies have also been discussed.

Exploring epitranscriptomics for crop improvement and environmental stress tolerance.

Climate change and stressful environmental conditions severely hamper crop growth, development and yield. Plants respond to environmental perturbations, through their plasticity provided by key-genes, governed at post-/transcriptional levels. Gene-regulation in plants is a multilevel process controlled by diverse cellular entities that includes transcription factors (TF), epigenetic regulators and non-coding RNAs beside others. There are successful studies confirming the role of epigenetic modifications (DNA-methylation/histone-modifications) in gene expression. Recent years have witnessed emergence of a highly specialized field the "Epitranscriptomics". Epitranscriptomics deals with investigating post-transcriptional RNA chemical-modifications present across the life forms that change structural, functional and biological characters of RNA. However, deeper insights on of epitranscriptomic modifications, with >140 types known so far, are to be understood fully. Researchers have identified epitranscriptome marks (writers, erasers and readers) and mapped the site-specific RNA modifications (m6A, m5C, 3' uridylation, etc.) responsible for fine-tuning gene expression in plants. Simultaneous advancement in sequencing platforms, upgraded bioinformatic tools and pipelines along with conventional labelled techniques have further given a statistical picture of these epitranscriptomic modifications leading to their potential applicability in crop improvement and developing climate-smart crops. We present herein the insights on epitranscriptomic machinery in plants and how epitranscriptome and epitranscriptomic modifications underlying plant growth, development and environmental stress responses/adaptations. Third-generation sequencing technology, advanced bioinformatics tools and databases being used in plant epitranscriptomics are also discussed. Emphasis is given on potential exploration of epitranscriptome engineering for crop-improvement and developing environmental stress tolerant plants covering current status, challenges and future directions.

Genus Barleria L. (Acanthaceae): a review of its taxonomy, cytogenetics, phytochemistry and pharmacological potential.

OBJECTIVES: Barleria, a large genus of the Acanthaceae family, comprises more than 300 species with diverse taxonomy, cytogenetics, phytochemistry and pharmacological potential. Therefore, the aim of this review is to critically assess the research on Barleria and provide guidance for future investigations. METHODS: The data were obtained from different sources, such as books, theses, journals and some of the websites and internet-based searches, published from 1901 to 2020. Data obtained from PubMed, Google Scholar, ScienceDirect, online electronic journals, SpringerLink, Wiley, etc. have also been used. KEY FINDINGS: The species of this genus exhibit considerable medicinal properties. Cytogenetical data are scantily available with chromosome counts available for only 24 species. The most common chromosome number is 2n = 2x = 40. So far, 187 compounds are reported from Barleria species. The active principles, their uses, toxicity and pharmacological effects are discussed. Essential oils, flavones, flavonoids, glycosides, terpenes and terpenoids form the major compounds. SUMMARY: It is highly recommended that the pharmacological and economic potential of Barleria species should be exploited and more detailed studies and attention be geared towards its utilization and conservation. In addition, to ensure maximum pharmacological benefits and sustainable use, it is necessary to have empirical information explaining its ethnobotanical values as well as commercial potential.

MicroRNA-mediated bioengineering for climate-resilience in crops.

Global projections on the climate change and the dynamic environmental perturbations indicate severe impacts on food security in general, and crop yield, vigor and the quality of produce in particular. Sessile plants respond to environmental challenges such as salt, drought, temperature, heavy metals at transcriptional and/or post-transcriptional levels through the stress-regulated network of pathways including transcription factors, proteins and the small non-coding endogenous RNAs. Amongs these, the miRNAs have gained unprecedented attention in recent years as key regulators for modulating gene expression in plants under stress. Hence, tailoring of miRNAs and their target pathways presents a promising strategy for developing multiple stress-tolerant crops. Plant stress tolerance has been successfully achieved through the over expression of microRNAs such as Os-miR408, Hv-miR82 for drought tolerance; OsmiR535A and artificial DST miRNA for salinity tolerance; and OsmiR535 and miR156 for combined drought and salt stress. Examples of miR408 overexpression also showed improved efficiency of irradiation utilization and carbon dioxide fixation in crop plants. Through this review, we present the current understanding about plant miRNAs, their roles in plant growth and stress-responses, the modern toolbox for identification, characterization and validation of miRNAs and their target genes including in silico tools, machine learning and artificial intelligence. Various approaches for up-regulation or knock-out of miRNAs have been discussed. The main emphasis has been given to the exploration of miRNAs for development of bioengineered climate-smart crops that can withstand changing climates and stressful environments, including combination of stresses, with very less or no yield penalties.

Nitric oxide, crosstalk with stress regulators and plant abiotic stress tolerance.

KEY MESSAGE: Nitric oxide is a dynamic gaseous molecule involved in signalling, crosstalk with stress regulators, and plant abiotic-stress responses. It has great exploratory potentials for engineering abiotic stress tolerance in crops. Nitric oxide (NO), a redox-active gaseous signalling molecule, though present uniformly through the eukaryotes, maintain its specificity in plants with respect to its formation, signalling, and functions. Its cellular concentrations are decisive for its function, as a signalling molecule at lower concentrations, but triggers nitro-oxidative stress and cellular damage when produced at higher concentrations. Besides, it also acts as a potent stress alleviator. Discovered in animals as neurotransmitter, NO has come a long way to being a stress radical and growth regulator in plants. As a key redox molecule, it exhibits several key cellular and molecular interactions including with reactive chemical species, hydrogen sulphide, and calcium. Apart from being a signalling molecule, it is emerging as a key player involved in regulations of plant growth, development and plant-environment interactions. It is involved in crosstalk with stress regulators and is thus pivotal in these stress regulatory mechanisms. NO is getting an unprecedented attention from research community, being investigated and explored for its multifaceted roles in plant abiotic stress tolerance. Through this review, we intend to present the current knowledge and updates on NO biosynthesis and signalling, crosstalk with stress regulators, and how biotechnological manipulations of NO pathway are leading towards developing transgenic crop plants that can withstand environmental stresses and climate change. The targets of various stress responsive miRNA signalling have also been discussed besides giving an account of current approaches used to characterise and detect the NO.

Severe thrombocytopenia in tetralogy of Fallot patients: A contraindication for corrective surgery?

A 3-year-old boy with tetralogy of Fallot and recurrent cyanotic spells was found to have severe thrombocytopenia with platelet counts in the range of 11-30,000/mm3. There was a hesitation to operate in view of the high bleeding risk due to profound thrombocytopenia. However, the total correction was done after excluding other causes of thrombocytopenia. His platelet count dramatically improved after the operation.

Ultrasound-Assisted Facile Synthesis of Nanostructured Hybrid Vesicle for the Nasal Delivery of Indomethacin: Response Surface Optimization, Microstructure, and Stability.

This work is devoted to design a novel nanostructured hybrid vesicle (NHV) made of lecithin and an acrylate/C10-C30 alkyl acrylate for the nasal delivery of a model active indomethacin (IND), and further to probe its microstructure, intermolecular interactions, drug release behavior, ex vivo permeation, and stability. NHVs were prepared by cavitation technology employing RSM-based central composite design (CCD). Amount of lecithin (X1), power of ultrasound (X2), and sonication time (X3) were selected as three independent variables while the studied response included Z-Avg (nm), polydispersity index (PDI), and zeta potential (mV). The designed system (NHV) was investigated through dynamic (DLS) and electrophoretic light scattering (ELS), attenuated total reflectance (ATR-FTIR), oscillatory measurement (stress and frequency sweep), and transmission electron microscopy (TEM). CCD was found useful in optimizing NHV. An optimized formulation (S6) had Z-Avg 80 nm, PDI 0.2, and zeta potential of - 43.26 mV. Morphology investigation revealed spherical vesicles with smaller TEM diameters (the largest particle being 52.26 nm). ATR analysis demonstrated significant intermolecular interactions among the drug (IND) and the components of vesicles. The designed vesicles had an elastic predominance and displayed supercase II (n > 1) type of drug release. Besides, the vesicles possessed potential to transport IND across the nasal mucosa with the steady-state flux (µg/cm2/h) and permeability coefficient (cm/h) of 26.61 and 13.30 × 10-3, respectively. NHV exhibited an exceptional stability involving a combination of electrostatic and steric interactions while the histopathology investigation confirmed their safety for nasal administration.

Collagen Promotes Higher Adhesion, Survival and Proliferation of Mesenchymal Stem Cells.

Mesenchymal stem cells (MSC) can differentiate into several cell types and are desirable candidates for cell therapy and tissue engineering. However, due to poor cell survival, proliferation and differentiation in the patient, the therapy outcomes have not been satisfactory. Although several studies have been done to understand the conditions that promote proliferation, differentiation and migration of MSC in vitro and in vivo, still there is no clear understanding on the effect of non-cellular bio molecules. Of the many factors that influence the cell behavior, the immediate cell microenvironment plays a major role. In this context, we studied the effect of extracellular matrix (ECM) proteins in controlling cell survival, proliferation, migration and directed MSC differentiation. We found that collagen promoted cell proliferation, cell survival under stress and promoted high cell adhesion to the cell culture surface. Increased osteogenic differentiation accompanied by high active RHOA (Ras homology gene family member A) levels was exhibited by MSC cultured on collagen. In conclusion, our study shows that collagen will be a suitable matrix for large scale production of MSC with high survival rate and to obtain high osteogenic differentiation for therapy.

Characterization of MEMS piezoresistive pressure sensors using AFM.

Atomic force microscope (AFM) is adapted to characterize an ultrasensitive piezoresistive pressure sensor based on microelectromechanical system (MEMS) technology. AFM is utilized in contact mode to exert force on several different micromachined diaphragm structures using a modified silicon cantilever with a particle attached to its end. The applied force is adjusted by changing the trigger voltage during each engage step of the probe-tip on the diaphragm surface. The contact force is determined from the force plots obtained for each trigger voltage in advanced force mode. Low force values in the range of 0.3-5muN have been obtained with this method. This force induces strain on the bridge-arm of the diaphragm where the polysilicon resistor is located. The resultant change in the resistance produced due to varying force/pressure is measured using a delta mode current-voltage (I-V) measurement set-up. The contact mode AFM in conjunction with a nanovoltmeter enables the calibration of very sensitive force sensors down to 0.3muN.