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Objective: In the present study is investigated of phytochemicals and antioxidant activities of the leaf extracts from Meliosma simplicifolia (L.). Methods: The seaweed sample was subjected to extraction with solvents of different polarities (hot water, methanol, acetone, chloroform, and petroleum ether) and screened for phytochemicals according to standard methods. The ability of the plant extract to act as hydrogen/electrons donor or scavenger of radicals was determined by in vitro antioxidant assays using 2,2-diphenyl-2-picryl-hydrazyl free radical (DPPH) scavenging, reducing power assay, and superoxide radical (O2•) scavenging activity. The ICPMS and GC-MS analysis of the methanol leaf extract of M. simplicifolia was revealed the presence of antibacterial. Results: Phytochemical analysis revealed the presence of Preliminary metabolites Quantitative studies of estimated phenol, flavonoid and tannin, as for the methanol extract of stem showed the highest content of phenolic compounds (40.71±0.94 GAE mg/100). Antioxidant activities were concluded the estimation M. simplicifolia leaf for as followed the studies. In leaf the highest DPPH scavenging activity (132.3 μg/ml), ICP-MS analysis of the leaf extract showed the presence minerals such as Mg, Fe, Cu, Na, and Zn in excess. The leaf extract of the plant was also tested for its antibacterial activity and results confirmed that it has potential activity. Conclusion: The preliminary studies in the methanol extract of the leaf of M. simplicifolia are suggestive of the antibacterial potentials of the methanol extract of leaves of M. simplicifolia.

The anticorrosive activity of Combretum indicum (CI) leaf extract has been examined on mild steel corrosion in 1M HCl by using weight loss and electrochemical measurement techniques. The results of these experimental techniques have consistent in confirming the effect of the CI leaf extract on corrosion inhibition and a mechanism for the inhibition was proposed. The dissolution rate of steel in acidic solution was found to be sensitive to the concentration of CI leaf extract and the corrosion rate reduces when the concentration of CI leaf extract is increased and reaches the high protection efficiency. The potentiodynamic polarization study confirms the mixed mode of inhibition. Ultraviolet–visible (UV) and Fourier -transform infrared (FTIR) spectroscopic techniques were performed to evaluate the phenomenon of adsorption in the plant extract and the functional groups. The surface morphology analysis (SEM) further corroborates the adsorption of inhibitor molecules over the metal surface.

Erythropoietic protoporphyria (EPP) is a rare disease in which patients experience severe light sensitivity. It is caused by a deficiency of ferrochelatase (FECH), the last enzyme in heme biosynthesis (HBS). The lack of FECH causes accumulation of its photoreactive substrate protoporphyrin IX (PPIX) in patients' erythrocytes. Here, we explored an approach for the treatment of EPP by decreasing PPIX synthesis using small-molecule inhibitors directed to factors in the HBS pathway. We generated a FECH-knockout clone from K562 erythroleukemia cells, which accumulates PPIX and undergoes oxidative stress upon light exposure. We used these matched cell lines to screen a set of publicly available inhibitors of factors in the HBS pathway. Inhibitors of the glycine transporters GlyT1 and GlyT2 lowered levels of PPIX and markers of oxidative stress selectively in K56211B4 cells, and in primary erythroid cultures from an EPP patient. Our findings open the door to investigation of glycine transport inhibitors for HBS disorders.

Humic substances (HS) and humic acids (HA) are proven to enhance nutrient uptake and growth in plants. Foliar application of urea combined with HS and HA offers an alternative strategy to increase nitrogen use efficiency (NUE). The objective of this study was to understand the effects of foliar application of HA and HS along with urea on NUE and response of different biometric, biochemical and physiological traits of sugarcane with respect to cultivar, mode of foliar application, geographic location and intervals of foliar application. To study this, two different independent Experiments were conducted in green house facilities at two different agro-climatic zones (USA and Brazil) using two different predominant varieties, modes and intervals of foliar applications. The three different foliar applications used in this study were (1) urea (U), (2) mixture of urea and HS (U+HS) and (3) HA (U+HA). In both Experiments, 15N (nitrogen isotope) recovery or NUE was higher in U+HS followed by U+HA. However, magnitude of NUE changed according to the differences in two Experiments. Results showed that foliar application of U+HS and U+HA was rapidly absorbed and stored in the form of protein and starch. Also induced changes in photosynthesis, intrinsic water use efficiency, protein, total soluble sugars and starch signifying a synergistic effect of U+HS and U+HA on carbon and nitrogen metabolism. These results showed promising use of HS and HA with urea to improve NUE in sugarcane compared to using the urea alone. Simultaneously, mode, quantity, and interval of foliar application should be standardized based on the geographic locations and varieties to optimize the NUE., Sugarcane; Urea; Humic substance; Humic acid; Nitrogen use efficiency; Agricultural water management; Agronomy; Crop biomass; Crop breeding; Crop production; Crop protection; Crop quality; Crop yields; Field crops; Agriculture, Plant biology.

Erythropoietic protoporphyria (EPP) is a rare genetic disease in which patients experience acute phototoxic reactions after sunlight exposure. It is caused by a deficiency in ferrochelatase (FECH) in the heme biosynthesis pathway. Most patients exhibit a loss-of-function mutation in trans to an allele bearing a SNP that favors aberrant splicing of transcripts. One viable strategy for EPP is to deploy splice-switching oligonucleotides (SSOs) to increase FECH synthesis, whereby an increase of a few percent would provide therapeutic benefit. However, successful application of SSOs in bone marrow cells is not described. Here, we show that SSOs comprising methoxyethyl-chemistry increase FECH levels in cells. We conjugated one SSO to three prototypical targeting groups and administered them to a mouse model of EPP in order to study their biodistribution, their metabolic stability and their FECH splice-switching ability. The SSOs exhibited distinct distribution profiles, with increased accumulation in liver, kidney, bone marrow and lung. However, they also underwent substantial metabolism, mainly at their linker groups. An SSO bearing a cholesteryl group increased levels of correctly spliced FECH transcript by 80% in the bone marrow. The results provide a promising approach to treat EPP and other disorders originating from splicing dysregulation in the bone marrow., Nucleic Acids Research, 48 (9), ISSN:1362-4962, ISSN:0301-5610

Biological nitrogen fixation (BNF) in soybean [Glycine max (L.) Merr.] plays an important role in sustainable agriculture, reducing the limitations associated with other sources of N such as fertilizers and soils. Our major objective was to evaluate the weekly pattern of BNF in soybean influenced by nodule formation, using three different laboratory‐cultured Bradyrhizobium strains. Plants were grown in 1‐m polyvinyl chloride (PVC) columns for 17 week in a greenhouse and BNF was determined using an integrated approach by assessing nodule formation, stem ureide‐N, and N partitioning in plant parts. Bradyrhizobium strains showed overall similar plant performance and N2−fixation capacity. During the beginning of flowering/full bloom (R1/R2) growth stages, nodule formation significantly increased and reached a maximum at pod‐filling (R4) stage. Stem ureide‐N was detected at early growth stages even with fewer small nodules, which significantly increased after the beginning of pod formation (R3). Peak N2−fixing rate (g N kg−1 d−1) started to decline after the onset of seed filling (R5.5). Relationship between BNF and nodule parameters (nodule number [R2 = .65] and nodule weight [R2 = .62]) suggested that they can be used as predictors of BNF.

Duchenne muscular dystrophy (DMD) is a lethal muscle-wasting disease caused by the lack of dystrophin in muscle fibers that is currently without curative treatment. Mesoangioblasts (MABs) are multipotent progenitor cells that can differentiate to a myogenic lineage and that can be used to express Dystrophin upon transplantation into muscles, in autologous gene therapy approaches. However, their fate in the muscle environment remains poorly characterized. Here, we investigated the differentiation fate of MABs following their transplantation in DMD murine muscles using a mass cytometry strategy. This allowed the identification and isolation of a fraction of MAB-derived cells presenting common properties with satellite muscle stem cells. This analysis also indicated that most cells did not undergo a myogenic differentiation path once in the muscle environment, limiting their capacity to restore dystrophin expression in transplanted muscles. We therefore assessed whether MAB treatment with cytokines and growth factors prior to engraftment may improve their myogenic fate. We identified a combination of such signals that ameliorates MABs capacity to undergo myogenic differentiation in vivo and to restore dystrophin expression upon engraftment in myopathic murine muscles., Stem Cell Research, 41, ISSN:1873-5061, ISSN:1876-7753

Deficiency in ferrochelatase (FECH), the last enzyme in the heme biosynthetic pathway, leads to an accumulation of protoporphyrin IX (PPIX) that causes a severely painful phototoxic reaction of the skin in patients with erythropoietic protoporphyria (EPP). Besides phototoxicity of the skin, EPP patients often present with symptoms of iron deficiency in form of a microcytic and hypochromic anemia with low serum iron and ferritin. In addition, elevated aminolevulinic acid synthase 2 (ALAS2) both at the mRNA and protein levels have been observed among EPP patients. ALAS is the first enzyme in the pathway and exists in two isoforms, whereby the isoform 2 (ALAS2) is expressed exclusively in erythropoiesis. The mRNA of ALAS2 contains an iron response element (IRE) at its 5′UTR. When iron is limited, iron response element binding protein 2 (IRP2) binds to the IRE of ALAS2 mRNA and suppresses its translation. In this study, we demonstrated that iron deprivation increased the amount of ALAS2 mRNA as well as the ratio of ALAS2 to FECH mRNAs in cultured erythroleukemic K562 cells. At the protein level, however, iron deprivation in the cell line caused reductions in both enzymes as shown by the Western blot analysis. A comparable increase in the ratio of ALAS2 to FECH mRNAs was also found in EPP patients indicating an imbalance in heme biosynthesis. As iron cannot be completely missing from an organism, we assume that in EPP patients, a certain amount of ALAS2 mRNA is translated despite a partial deficiency of FECH. The increase in ALAS2 enzyme contributes to the accumulation in PPIX in the patients. Targeted inhibition of ALAS2 could therefore be a treatment option for EPP., Molecular Genetics and Metabolism, 128 (3), ISSN:1096-7192, ISSN:1096-7206

In the original version of this article unfortunately column six and seven in Table 2 were wrong due to typesetting mistakes.

Duchenne muscular dystrophy (DMD) is a lethal muscle-wasting disease currently without cure. We investigated the use of the PiggyBac transposon for full-length dystrophin expression in murine mesoangioblast (MABs) progenitor cells. DMD murine MABs were transfected with transposable expression vectors for full-length dystrophin and transplanted intramuscularly or intra-arterially into mdx/SCID mice. Intra-arterial delivery indicated that the MABs could migrate to regenerating muscles to mediate dystrophin expression. Intramuscular transplantation yielded dystrophin expression in 11%–44% of myofibers in murine muscles, which remained stable for the assessed period of 5 months. The satellite cells isolated from transplanted muscles comprised a fraction of MAB-derived cells, indicating that the transfected MABs may colonize the satellite stem cell niche. Transposon integration site mapping by whole-genome sequencing indicated that 70% of the integrations were intergenic, while none was observed in an exon. Muscle resistance assessment by atomic force microscopy indicated that 80% of fibers showed elasticity properties restored to those of wild-type muscles. As measured in vivo, transplanted muscles became more resistant to fatigue. This study thus provides a proof-of-principle that PiggyBac transposon vectors may mediate full-length dystrophin expression as well as functional amelioration of the dystrophic muscles within a potential autologous cell-based therapeutic approach of DMD., Molecular Therapy, 26 (4), ISSN:1525-0016, ISSN:1525-0024

An emerging therapeutic approach for Duchenne muscular dystrophy is the transplantation of autologous myogenic progenitor cells genetically modified to express dystrophin. The use of this approach is challenged by the difficulty in maintaining these cells ex vivo while keeping their myogenic potential, and ensuring sufficient transgene expression following their transplantation and myogenic differentiation in vivo. We investigated the use of the piggyBac transposon system to achieve stable gene expression when transferred to cultured mesoangioblasts and into murine muscles. Without selection, up to 8% of the mesoangioblasts expressed the transgene from 1 to 2 genomic copies of the piggyBac vector. Integration occurred mostly in intergenic genomic DNA and transgene expression was stable in vitro. Intramuscular transplantation of mouse Tibialis anterior muscles with mesoangioblasts containing the transposon led to sustained myofiber GFP expression in vivo. In contrast, the direct electroporation of the transposon-donor plasmids in the mouse Tibialis muscles in vivo did not lead to sustained transgene expression despite molecular evidence of piggyBac transposition in vivo. Together these findings provide a proof-of-principle that piggyBac transposon may be considered for mesoangioblast cell-based therapies of muscular dystrophies.

A cornerstone of autologous cell therapy for Duchenne muscular dystrophy is the engineering of suitable cells to express dystrophin in a stable fashion upon differentiation to muscle fibers. Most viral transduction methods are typically restricted to the expression of truncated recombinant dystrophin derivatives and by the risk of insertional mutagenesis, while non-viral vectors often suffer from inefficient transfer, expression and/or silencing. Here we addressed such limitations by using plasmid vectors containing nuclear matrix attachment regions (MAR). Using in vitro transfection and intra muscular transplantation in nude and immunosuppressed mdx mice, we show that clones of mesoangioblast skeletal muscle progenitors can be generated to mediate stable expression from MAR-containing vectors, while maintaining their ability to differentiate in vitro and in vivo and to express dystrophin after transplantation in dystrophic mouse muscles. We conclude that the incorporation of MARs into plasmid vectors may improve non-viral plasmid-based cell therapy feasibility.

Relative quantification ofalgCgene expression was evaluated in the multidrug resistant strainAcinetobacter baumanniiAIIMS 7 biofilm (3 to 96 h, on polystyrene surface) compared to the planktonic counterparts. Comparison revealed differentialalgCexpression pattern with maximum 81.59-fold increase in biofilm cells versus 3.24-fold in planktonic cells (P<0.05). Expression levels strongly correlated with specific biofilm stages (scale of 3 to 96 h), coinciding maximum at initial surface attachment stage (9 h) and biofilm maturation stage (48 h). Cloning, heterologous expression, and bioinformatics analyses indicatedalgCgene product as the bifunctional enzyme phosphomannomutase/phosphoglucomutase (PMM/PGM) of ∼53 kDa size, which augmented biofilms significantly inalgCclones compared to controls (lackingalgCgene), further localized by scanning electron microscopy. Moreover, molecular dynamics analysis on the three-dimensional structure of PMM/PGM (simulated up to 10 ns) revealed enzyme structure as stable and similar to that inP. aeruginosa(synthesis of alginate and lipopolysaccharide core) and involved in constitution of biofilm EPS (extracellular polymeric substances). Our observation on differential expression pattern ofalgChaving strong correlation with important biofilm stages, scanning electron-microscopic evidence of biofilm augmentation taken together with predictive enzyme functions via molecular dynamic (MD) simulation, proposes a new basis ofA. baumanniiAIIMS 7 biofilm development on inanimate surfaces.

Metallochlorophyllin, a water soluble analogue of the ubiquitous green pigment chlorophyll belongs to a group of compounds, porphyrins that contain a cheated metal ion in the center of the molecule. Tin chlorophyll in is the metallochlorophyllin with tin as its central metal ion and the phytol chains lost. The aim of the present study was to explore the antimicrobial activity of tin chlorophyll in from the leaves of Morinda citrifolia L. The leaves were collected and the tin chlorophyll in was extracted. The extracted tin chlorophyll in was subjected for antimicrobial study by disc diffusion method. The test organisms include Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Salmonella typhi, Pseudomonas aeroginosa, Aspergillus niger, Rhizopus microsporus. The inhibition zones were detected and were calculated for the various organisms. The Minimal Inhibitory Concentration (MIC) was also determined and found to be 125µg/ml (Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeroginosa) and 250µg/ml (Escherichia coli, Salmonella typhi). The results of the present study conclude that tin chlorophyll in from the leaves of Morinda citrifolia L. showed significant antimicrobial activity.

Plants are the richest source of bioactive compounds and they have been used as medicine also. Chlorophyllin, a water soluble derivative of chlorophyll (chl) in which magnesium has been replaced with copper and the phytol chains lost. Chlorophyllin has been used by human population for over 50 years for medicinal purposes with no adverse effects. Chlorophyllin is a promising chemopreventive agent to block cancer primarily by inhibiting carcinogen such as AFB1. The objective was to extract the bioactive pigment chloropyllin from Rhinacanthus nasutus and characterize by IR and NMR and evaluate the effect of chlorophyllin on inhibition of cell proliferation in HepG2 and HeLa cell lines. The HepG2 and HeLa cell lines was exposed with different concentrations of chlorophyllin, and found that there was a significant dose dependent reduction in cell viability. The IC50 value at 48Hrs was 62.5µg/ml for Rhinacanthus nasutus Chlorophyllin on HepG2 Cell line and HeLa cell line. This is the first report of chlorophyllin from fresh leaves of Rhinacanthus nasutus on HepG2 and HeLa cell lines. It also suggests the benefit of these natural compounds against HepG2 and HeLa cell lines.

Medicinal plants are the rich source of harmless medicines, and used for the treatment of various diseases for thousands of years. They can provide biologically active molecules and lead structures for development of modified derivatives with enhanced activity or reduced activity. Green plants synthesize and preserve a variety of biochemical products, many of which are extractable and used as chemical feed stocks or as raw material for several scientific investigations. Tin chlorophyllin, a water soluble sodium salt of Chlorophyll is one among the family of phytochemical compounds. The central metal atom, magnesium of chlorophyll has been replaced with tin and the phytol chains lost in chlorophyllin. The antioxidant activity of tin chlorophyllin from the leaves of Morinda citrifolia L. was investigated by using in vitro antioxidant models such as DPPH scavenging assay and Phosphomolybdenum method. The free radical scavenging activity of tin chlorophyllin on the stable radical 1, 1-diphenyl-2-picrylhydrazyl (DPPH) was evaluated and the percentage of inhibition was determined. By phosphomolybdenum method, the total antioxidant capacity of tin chlorophyllin was determined and the absorbance values were expressed as the number of gram equivalent of ascorbic acid. The results demonstrate the potential antioxidant activity of tin chlorophyllin and support its possible role in human health protection and disease prevention.

Release of extracellular DNA (eDNA) was observed duringin vitrogrowth of a clinical strain ofAcinetobacter baumannii. Membrane vesicles (MV) of varying diameter (20–200 nm) containing DNA were found to be released by transmission electron microscopy (TEM) and atomic force microscopy (AFM). An assessment of the characteristics of the eDNA with respect to size, digestion pattern by DNase I/restriction enzymes, and PCR-sequencing, indicates a high similarity with genomic DNA. Role of eDNA in static biofilm formed on polystyrene surface was evaluated by biofilm augmentation assay using eDNA available in different preparations, for example, whole cell lysate, cell-free supernatant, MV suspension, and purified eDNA. Biofilm augmentation was seen up to 224.64%, whereas biofilm inhibition was 59.41% after DNase I treatment: confirming that eDNA facilitates biofilm formation inA. baumannii. This is the first paper elucidating the characteristics and role of eDNA inA. baumanniibiofilm, which may provide new insights into its pathogenesis.