Near-Infrared Fluorescent Carbon Nanotube Sensors for the Plant Hormone Family Gibberellins.

Gibberellins (GAs) are a class of phytohormones, important for plant growth, and very difficult to distinguish because of their similarity in chemical structures. Herein, we develop the first nanosensors for GAs by designing and engineering polymer-wrapped single-walled carbon nanotubes (SWNTs) with unique corona phases that selectively bind to bioactive GAs, GA3 and GA4, triggering near-infrared (NIR) fluorescence intensity changes. Using a new coupled Raman/NIR fluorimeter that enables self-referencing of nanosensor NIR fluorescence with its Raman G-band, we demonstrated detection of cellular GA in Arabidopsis, lettuce, and basil roots. The nanosensors reported increased endogenous GA levels in transgenic Arabidopsis mutants that overexpress GA and in emerging lateral roots. Our approach allows rapid spatiotemporal detection of GA across species. The reversible sensor captured the decreasing GA levels in salt-treated lettuce roots, which correlated remarkably with fresh weight changes. This work demonstrates the potential for nanosensors to solve longstanding problems in plant biotechnology.

Single-Crystal 2D Covalent Organic Frameworks for Plant Biotechnology.

Molecules chemically synthesized as periodic two-dimensional (2D) frameworks via covalent bonds can form some of the highest-surface area and -charge density particles possible. There is significant potential for applications such as nanocarriers in life sciences if biocompatibility can be achieved; however, significant synthetic challenges remain in avoiding kinetic traps from disordered linking during 2D polymerization of compatible monomers, resulting in isotropic polycrystals without a long-range order. Here, we establish thermodynamic control over dynamic control on the 2D polymerization process of biocompatible imine monomers by minimizing the surface energy of nuclei. As a result, polycrystal, mesocrystal, and single-crystal 2D covalent organic frameworks (COFs) are obtained. We achieve COF single crystals by exfoliation and minification methods, forming high-surface area nanoflakes that can be dispersed in aqueous medium with biocompatible cationic polymers. We find that these 2D COF nanoflakes with high surface area are excellent plant cell nanocarriers that can load bioactive cargos, such as the plant hormone abscisic acid (ABA) via electrostatic attraction, and deliver them into the cytoplasm of intact living plants, traversing through the cell wall and cell membrane due to their 2D geometry. This synthetic route to high-surface area COF nanoflakes has promise for life science applications including plant biotechnology.

Decoding the Assembly of Mixed and Branched Heterotypic Ubiquitin Chains.

Ubiquitination is a post-translational modification that regulates cell signaling, immune response, protein processing, molecular trafficking, and DNA repair. Generally, molecular trafficking and DNA repair processes need the attachment of a single ubiquitin on a substrate, known as monoubiquitination. The other functions of ubiquitin require the assembly of polymeric ubiquitin chains on the substrate, known as polyubiquitination. The chains are linked through the lysine residues of ubiquitin, and depending on which lysine is connected, the chains could be heterotypic or homotypic. Heterotypic polyubiquitin chains can be mixed, branched, or combined, generating myriad cellular signals with functions distinct from the homotypic ubiquitin chains. The molecular rules of heterotypic chain assembly are poorly understood due to the lack of techniques to monitor their assembly. New approaches are required to monitor the conjugation site of new ubiquitin molecules on a pre-existing chain. Here, we describe a new method based on isotopic labeling and mass spectrometry to study the assembly of heterotypic chains called isotopically resolved mass spectrometry of peptides (IRMSP). The technique is demonstrated using multiple ubiquitin enzymes and ubiquitin chains as substrates. It causes minimal perturbation to the enzyme/substrate and will be instrumental in studying the assembly of large polymeric ubiquitin chains. Using this technique, it is feasible to monitor how and with what rate branched ubiquitin chains grow in different directions in a single experiment.

Japanese Encephalitis Virus Infection Causes an Imbalance in the Activation of Mitochondrial Fusion/Fission Genes and Triggers the Activation of NOX2-mediated Oxidative Stress and Neuronal Cell Death.

Mitochondria dysfunction may be an important contributor to Japanese encephalitis (JE) viral infection disease pathogenesis. In the current study, we define whether changes in mitochondrial DNA copy number (which is the biomarker for mitochondrial function) and alteration in mitochondria dynamics (fusion and fission) contribute to the pathology of the JE virus in vivo mice model. We found decreased mitochondria copy number, reduced activation of mitochondrial fission (FIS1/DRP1), and increased activation of mitochondrial fusion (MFN1/MFN2/OPA1) genes that are associated with increased NOX2-mediated ROS generation and neuronal cell death following JE virus infection. Furthermore, we found that antioxidant glutathione level decreases. In summary, the following study demonstrates that JE viral infection causes an imbalance in mitochondrial fission/fusion gene activation and promotes NOX2-mediated oxidative stress and cell death, suggesting that intervention in mitochondrial dynamics might be a potential therapeutic strategy for combating oxidative stress and inflammatory process in JE viral infection.

Quantifying Agglomerate-to-Wall Impaction in Dry Powder Inhalers.

PURPOSE: The probability of agglomerate-to-wall collision was quantified using a unique image processing technique applied to high-speed microscopic images. The study aimed to investigate the effects of flow rate and particle size on the percentage of colliding agglomerates detected within an in-house powder dispersion device. METHOD: The device consists of a swirl chamber and two tangential inlets in various configurations, designed to emulate the geometric features of commercial devices such as the Aerolizer® and Osmohaler®. The test cases were conducted with constant flow rates of 30 SLPM and 60 SLPM. Four powder samples were tested, including carrier Respitose® SV010 (median volume diameter 104 µm, span 1.7) and mannitol of three constituent primary particle sizes (3 µm, 5 µm and 7 µm; span 1.6 - 1.9). RESULTS: At the lower flow rate of 30 SLPM, collision frequencies were significantly different between powders of different constituent particle sizes, but the effects of powder properties diminished on increasing the flow rate to 60 SLPM. At the higher flow rate, all powders experienced a significant increase in the proportion of colliding particles. CONCLUSION: Analysis of collision events showed that the probability of collision for each agglomerate increased with agglomerate diameter and velocity. Experimental data of agglomerate-to-wall collision were utilised to develop a logistic regression model that can accurately predict collisions with various powders and flow rates.

Employing algal biomass for fabrication of biofuels subsequent to phytoremediation.

An alga belongs to the multi-pertinent group which can add to a significant sector of environment. They show a prevailing gathering of microorganisms for bioremediation due to their significant capacity to inactivate toxic heavy metals. It can easily absorb or neutralize the toxicity of heavy metals from water and soil through phytoremediation. Biosorption is a promising innovation that focuses on novel, modest, and exceptionally successful materials to apply in phytoremediation technology. Furthermore, algal biomass can be used for biofuel generation after phytoremediation using thermochemical or biological transformation processes. The algal components get affected by heavy metals during phytoremediation, but with the help of different techniques, these are yield efficient. The extreme lipid and mineral substances of microalgae have been proven helpful for biofuel manufacturing and worth extra products. Biofuels produced are bio-oil, biodiesel, bioethanol, biogas, etc. The reuse capability of algae can be utilized toward ecological manageability and economic facility. In this review article, the reuse and recycling of algal biomass for biofuel production have been represented. This novel technique has numerous benefits and produces eco-friendly and economically beneficial products.

This article explains how algal biomass left over after phytoremediation can be reused and recycled to produce biofuel. It is a novel technique that is cost-effective, demands less time for biofuel production as it does not include cultivation and harvesting time, and produces products that are economically and environmentally beneficial for society.

Synthetic azo dye bio-decolorization by Priestia sp. RA1: process optimization and phytotoxicity assessment.

Azo compounds represent the most diverse group of colorants widely employed in industrial sectors. Being highly toxic and recalcitrant compound, azo dyes pose a threat to plants, animals, and humans. In the present report, bio-decolorization of azo dye, reactive black 5, was evaluated by newly isolated Priestia sp. RA1. Strain RA1 was able to decolorize 97% of 100 ppm reactive black 5 in 60 h. Specific activity of dye decolorization was found to be 0.233 µmol min-1 g-1 dry cells. Successful decolorization over a broad range of pH, salinity, temperature, and initial dye concentration was observed. Phytotoxicity assay on agriculturally important crops showed considerable difference in percentage seed germination and growth when treated with original and bio-decolorized dye samples. Bio-decolorization at high dye concentrations, promising decolorization rate, and non-toxic nature of treated products suggest the potential of strain RA1 for bioremediation of dye-contaminated water and its re-use in the industries.

A theory of mechanical stress-induced H2O2 signaling waveforms in Planta.

Recent progress in nanotechnology-enabled sensors that can be placed inside of living plants has shown that it is possible to relay and record real-time chemical signaling stimulated by various abiotic and biotic stresses. The mathematical form of the resulting local reactive oxygen species (ROS) wave released upon mechanical perturbation of plant leaves appears to be conserved across a large number of species, and produces a distinct waveform from other stresses including light, heat and pathogen-associated molecular pattern (PAMP)-induced stresses. Herein, we develop a quantitative theory of the local ROS signaling waveform resulting from mechanical stress in planta. We show that nonlinear, autocatalytic production and Fickian diffusion of H2O2 followed by first order decay well describes the spatial and temporal properties of the waveform. The reaction-diffusion system is analyzed in terms of a new approximate solution that we introduce for such problems based on a single term logistic function ansatz. The theory is able to describe experimental ROS waveforms and degradation dynamics such that species-dependent dimensionless wave velocities are revealed, corresponding to subtle changes in higher moments of the waveform through an apparently conserved signaling mechanism overall. This theory has utility in potentially decoding other stress signaling waveforms for light, heat and PAMP-induced stresses that are similarly under investigation. The approximate solution may also find use in applied agricultural sensing, facilitating the connection between measured waveform and plant physiology.

Design and Development of a Bimodal Optical Instrument for Simultaneous Vibrational Spectroscopy Measurements.

Vibrational spectroscopy techniques are widely used in analytical chemistry, physics and biology. The most prominent techniques are Raman and Fourier-transform infrared spectroscopy (FTIR). Combining both techniques delivers complementary information of the test sample. We present the design, construction, and calibration of a novel bimodal spectroscopy system featuring both Raman and infrared measurements simultaneously on the same sample without mutual interference. The optomechanical design provides a modular flexible system for solid and liquid samples and different configurations for Raman. As a novel feature, the Raman module can be operated off-axis for optical sectioning. The calibrated system demonstrates high sensitivity, precision, and resolution for simultaneous operation of both techniques and shows excellent calibration curves with coefficients of determination greater than 0.96. We demonstrate the ability to simultaneously measure Raman and infrared spectra of complex biological material using bovine serum albumin. The performance competes with commercial systems; moreover, it presents the additional advantage of simultaneously operating Raman and infrared techniques. To the best of our knowledge, it is the first demonstration of a combined Raman-infrared system that can analyze the same sample volume and obtain optically sectioned Raman signals. Additionally, quantitative comparison of confocality of backscattering micro-Raman and off-axis Raman was performed for the first time.

Role of calcium phosphate and bioactive glass coating on in vivo bone healing of new Mg-Zn-Ca implant.

Present investigation focuses on development and detailed characterization of a new Mg alloy sample (BM) with and without coating of hydroxyapatite (BMH) and bioactive glass (BMG) by air plasma spray method. After detailed mechano-physico-chemical characterization of powders and coated samples, electrochemical corrosion and SBF immersion tests were carried out. Detailed in vitro characterizations for cell viability were undertaken using MG-63 cell line followed by in vivo tests in rabbit model for studying bone healing up to 60 days. Starting current density increases from BM to BMH to BMG indicating highest resistance towards corrosion in case of BMG samples, however BMH also showed highest icorr value suggesting slowest rate of corrosion than BM and BMG samples. Dissolution of calcium ion in case of BMH and BMG control formation of apatite phases on surface. Ca2+ ions of coatings and from SBF solution underwent reduction reaction simultaneously with conversion of Mg to MgCl2 releasing OH- in the solution, which increases pH. Viability and propagation of human osteoblast-like cells was verified using confocal microscopy observations and from expression of bone specific genes. Alkaline phosphatase assay and ARS staining indicate cell proliferation and production of neo-osseous tissue matrix. In vivo, based on histology of heart, kidney and liver, and immune response of IL-2, IL-6 and TNFα, all the materials show no adverse effects in body system. The bone creation was observed to be more for BMH. Although both BMH and BMG show rays of possibilities in early new bone formation and tough bone-implant bonding at interface as compared to bare Mg alloy, however, BMG showed better well-sprayed coating covering on substrate and resistance against corrosion prior implanting in vivo. Also, better apatite formation on this sample makes it more favourable implant.

Clinical Efficacy and Safety of Remdesivir among Hospitalised Adult Patients with RT PCR Confirmed COVID 19 Requiring ICU Care in Kalyana Karnataka.

BACKGROUND: Despite global efforts, COVID 19 pandemic is still posing a serious challenge with dearth of effective treatment options. Remdesivir has got FDA approval as the first COVID-19 anti-viral agent in adult and paediatric patients (aged ≥12 years and weighing at least 40 kg) requiring hospitalization. OBJECTIVE: The present prospective, observational, cross sectional study was planned to evaluate the impact of early initiation of Remdesivir on clinical outcomes in moderate to severe COVID 19 patients requiring ICU care. Materials and Methods: In this study, 100 consecutive symptomatic RT-PCR positive COVID 19 patients requiring admission to Intensive care unit based on predefined criteria were included for evaluation. All such moderate to severely ill patients were given Remdesivir intravenously as a 200-mg loading dose on day 1, followed by a 100-mg maintenance dose for the next four days along with other standard care. The main outcome measure analysed was the impact of early initiation of Remdesivir on recovery assessed by number of days in hospital, recovery, biochemical improvements and death. Results: Out of total 100 patients, 84 patients recovered with Remdesivir along with supportive treatment and were discharged from the hospital. Mean age of patients at presentation 53.5± 14.8 years with 5:1 male preponderance. Mean duration of hospital stay was 11.6 ± 5.9 days. D-dimer, CRP, Ferritin, IL-6 decreased significantly post treatment with P values <0.05, <0.001, <0.001, <0.01 respectively when compared to values at admission. No significant side effects were seen with remdesivir infusion. CONCLUSION: This real-world experience suggests that in the subset of hospitalized patients with moderate to severe pneumonia, early use of Remdesivir can lead to better clinical outcomes and help in reduction of associated mortality and morbidity of COVID-19.

Application of remote sensing in alpine grasslands cover mapping of western Himalaya, Uttarakhand, India.

Grasslands are the world's most extensive terrestrial ecosystem, which provides a variety of services for humans, such as carbon storage, food production, crop pollination, pest regulation, and are a major feed source for livestock. However, grasslands are today one of the most endangered ecosystems due to land-use change, agricultural intensification, land abandonment, as well as climate change. Grasslands are an integral part of human societies across the globe, which are broadly known as tropical savannah and temperate grasslands. In the Himalayan region, grasslands are found in more than 55% of the area and different climatic conditions lead to different varieties of grasslands like Danthonia grasslands, kobresia sedge meadow, etc. Grasslands deal with the spatial and temporal distribution of heterogeneous landscapes, which support a high diversity of various species. Owing to very rugged terrain and inaccessibility, the information on the extent of alpine grassland and percent grass cover (%) across the meadows is limited. Therefore, the present attempt was made to assess the current status of grassland in the alpine region of Uttarakhand above 3000 m asl. LANDSAT-8 (OLI and TIRS sensors) satellite data were used to delineate the grasslands using normalized difference vegetation indices (NDVIs) of the alpine region with the help of over 179 ground truth points out of which 50 points are testing points and 129 points are training points. Grass covers (%) were also assessed in the whole alpine region of Western Himalaya of Uttarakhand which nearly consists of over 75 meadows by using random plots (1 × 1 m, total 10 per site) in each meadow. Overall, 89.52% accuracy was achieved based on 50 randomly selected testing points. A total of 4949.25 sq. km area is under the different percentage of grass cover in the alpine region of Uttarakhand, Western Himalaya. Danthonia grasslands below 4000 m and Kobresia sedge meadows above 4000 m elevation are dominant in the state. In the alpine region, over 1056 sq. km grassland area have less than 10% grass cover indicating higher degraded and cold desert areas and only 565.69 sq. km area have more than 60% grass cover, which is highly favorable for rich biodiversity and grazing.

Impaired Autophagy Flux is Associated with Proinflammatory Microglia Activation Following Japanese Encephalitis Virus Infection.

Role of autophagy in Japanese encephalitis viral (JEV) infection is not well known. In the present study, we reported the role of autophagy flux in microglia activation, neurobehavioral function and neuronal death using a mouse model of JEV. Markers for autophagy (LC3-II/I, SQSTM1/P62, phos-Akt, phos-AMPK), and neuronal death (cleaved caspase 12, H2Ax, polyubiquitin) were investigated by western blot at 1, 3 and 7 days post inoculation. Cathepsin D was measured in cerebral cotex of JEV infected mice spectrophotometrically. Microglia activation and pro-inflammatory cytokines (IL1ß, TNF-α, IFNγ, IL6) were measured by immunohistochemistry, western blot and qPCR analysis. In order to determine the neuroinflammatory changes and autophagy mediated neuronal cell death, BV2-microglia and N2a-neuronal cells were used. Autophagy activation marker LC3-II/I and its substrate SQSTM1/P62 were significantly increased while cathepsin D activity was decreased on day 7 post inoculation in cerebral cortex. Microglia in cortex were activated and showed higher expression of proinflammatory mRNA of IL1ß, TNF-α, IFNγ and IL6, with increased DNA damage (H2AX) and neuronal cell death pathways in hippocampus and neurobehavioral dysfunction. Similar observations on JEV infection mediated autophagy flux inhibition and neuronal cell death was found in N2a neuronal cell. Collectively, our study provides evidence on the role of autophagy regulation, microglial activation and neurodegeneration following JEV infection.

Pyrrolidine ring puckering and prolyl amide bond configurations of 2-methyl-allo-hydroxyproline-based dipeptides.

An expeditious method for the synthesis of homo and heterochiral dipeptides containing l-alanine and d/l 2-methyl allo-hydroxyl prolines was developed using direct aminolysis of bicyclic lactones derived from d/l alanine. The impact of C-2 methylation and its spatial orientation on the pyrrolidine ring puckering and prolyl amide bond configuration was ascertained by solution NMR studies. The present studies reveal that C-2 methylation causes the prolyl amide bond to exist exclusively in the trans geometry in both homo- and heterochiral dipeptides. However, the spatial orientation of the C-2 methyl group and its i + 2 position in appropriately capped model dipeptides may nucleate into a turn like structure.

Correction to: Voltammetric detection of anti-HIV replication drug based on novel nanocomposite gold-nanoparticle-CaCO3 hybrid material.

The Figs. 2, 4 and 5 were published wrongly in this paper, due to inadvertent compilation of figures during uploading the paper.

Consumption of junk foods by school-aged children in rural Himachal Pradesh, India.

There has been an increase in the consumption of junk food (JF) among school-aged children (SAC) possibly leading to obesity and diet-related diseases among them. We do not have evidence on consumption of JF in rural areas; hence, we conducted a study to assess the consumption of JF by SAC in rural, Himachal Pradesh. A total of 425 children in the age group of 12-18 years studying in 30 government schools (clusters) were included. The clusters were selected using population proportionate to size sampling methodology. We found high prevalence (36%) of consumption of JF among SAC during the last 24 h. Efforts should be taken to reduce the consumption of JF by promotion of healthy dietary habits and educating children about the ill effects of JF.

AzaGly-Appended Peptidomimetics Structurally Related to PTR6154 as Potential PKB/Akt Inhibitors.

We report the synthesis and biological and physiochemical properties of a series of azaGly-appended peptidomimetics. We have developed a simple and facile synthesis for azapeptides on solid support without any side reaction. The azaGly is inserted by in situ reaction of disuccinimidyl carbonate with free amine followed by treatment of hydrazine hydrate at room temperature. The new series of peptidomimetics was prepared by azaGly scanning of heptapeptide Arg-Pro-Arg-Nle-Tyr-Dap-Nle (Akt-01), a GSK-3ß-derived Akt inhibitor. The azaGly-appended peptides showed significant improvement in biological activity and serum stability, with retention of conformation as evidenced by NMR and CD studies. The results clearly demonstrate that azaGly-appended peptides are new peptidomimetics. Their synthesis makes this approach highly useful for the development of novel peptidomimetics of therapeutic potential.

Effect of Subgingivally Delivered 10% Emblica officinalis Gel as an Adjunct to Scaling and Root Planing in the Treatment of Chronic Periodontitis - A Randomized Placebo-controlled Clinical Trial.

Emblica officinalis fruit possesses varied medicinal properties including cytoprotective antimicrobial, antioxidant, antiresorptive and antiinflammatory activity. The present study aimed to investigate the effect of subgingival application of indigenously prepared E. officinalis (Amla) sustained-release gel adjunctive to scaling and root planing (SRP) on chronic periodontitis. Forty-six patients (528 sites) were randomly assigned to control group (23;264): SRP +placebo gel and test group (23;264): SRP + 10% E. officinalis gel application. Periodontal parameters: plaque index, gingival index, probing pocket depth (PPD), clinical attachment level (CAL) and modified sulcus bleeding index (mSBI) were assessed at baseline, 2 and 3-month post-therapy. Forty patients (470 sites) completed the trial. When test and control sites were compared, significantly more reduction in mean PPD, mSBI, number of sites with PPD = 5-6 mm, PPD ≥ 7 mm, CAL ≥ 6 mm and greater CAL gain were achieved in test sites at 2- and 3-month post-therapy (p < 0.05). Locally delivered 10% E. officinalis sustained-release gel used as an adjunct to SRP may be more effective in reducing inflammation and periodontal destruction in patients with chronic periodontitis when compared with SRP alone. Copyright © 2016 John Wiley & Sons, Ltd.

Tetrahydrofuran amino acid-containing gramicidin S analogues with improved biological profiles.

Gramicidin S (GS) is a cyclic cationic antimicrobial peptide (CAP) with a wide spectrum of antibiotic activities whose usage has been limited to topical applications owing to its cytotoxic side effects. We have synthesized tetrahydrofuran amino acid (Taa)-containing GS analogues, and we have carried out conformational analysis and explored their structure activity relationships by evaluating their antitubercular, antibacterial and cytotoxic properties. Two of these analogues showed impressive as well as selective activity against Mycobacterium tuberculosis (MTB) without toxicity towards mammalian Vero cells or human RBCs, and are promising as potential leads.

Uterotonic use immediately following birth: using a novel methodology to estimate population coverage in four countries.

BACKGROUND: Postpartum hemorrhage (PPH) is the leading cause of maternal mortality in developing countries. While incidence of PPH can be dramatically reduced by uterotonic use immediately following birth (UUIFB) in both community and facility settings, national coverage estimates are rare. Most national health systems have no indicator to track this, and community-based measurements are even more scarce. To fill this information gap, a methodology for estimating national coverage for UUIFB was developed and piloted in four settings. METHODS: The rapid estimation methodology consisted of convening a group of national technical experts and using the Delphi method to come to consensus on key data elements that were applied to a simple algorithm, generating a non-precise national estimate of coverage of UUIFB. Data elements needed for the calculation were the distribution of births by location and estimates of UUIFB in each of those settings, adjusted to take account of stockout rates and potency of uterotonics. This exercise was conducted in 2013 in Mozambique, Tanzania, the state of Jharkhand in India, and Yemen. RESULTS: Available data showed that deliveries in public health facilities account for approximately half of births in Mozambique and Tanzania, 16% in Jharkhand and 24% of births in Yemen. Significant proportions of births occur in private facilities in Jharkhand and faith-based facilities in Tanzania. Estimated uterotonic use for facility births ranged from 70 to 100%. Uterotonics are not used routinely for PPH prevention at home births in any of the settings. National UUIFB coverage estimates of all births were 43% in Mozambique, 40% in Tanzania, 44% in Jharkhand, and 14% in Yemen. CONCLUSION: This methodology for estimating coverage of UUIFB was found to be feasible and acceptable. While the exercise produces imprecise estimates whose validity cannot be assessed objectively in the absence of a gold standard estimate, stakeholders felt they were accurate enough to be actionable. The exercise highlighted information and practice gaps and promoted discussion on ways to improve UUIFB measurement and coverage, particularly of home births. Further follow up is needed to verify actions taken. The methodology produces useful data to help accelerate efforts to reduce maternal mortality.