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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.

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.

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

Background Facial injuries due to flying windscreen glass pieces are common in road accident and require surgical intervention by plastic surgeons to debride, suture, and dress the injuries. Posthealing patients visit dermatologists for further treatment of residual scar tissue. Interestingly, tiny papules or small nodular scars seen on otherwise level background which persists for beyond 3 months (scar modeling time) may actually harbor small fragments of glass. Such lesions can only be treated by fractional CO2 therapy. Methods Two patients who had papules and small nodular lesions on the accident scarred areas were subjected to fractional CO2 laser (ultraPulse, Lumenis)with moderate fluence and high density, once and observed over a period of 12 weeks. Results On targeting the papules and small nodules, tiny particles of glass appeared out of the broken lesions. Discussion Irrespective of the surgery done on accident victims involving windshield glass injuries, fractional CO2 may be considered at a later stage, especially with presence of papules interspersed in the area of facial scars.

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

Duchenne muscular dystrophy (DMD) is still an incurable muscle degenerative disease; thus, numerous studies focused on novel therapeutic approaches. However, a simple assay of muscle function restoration remains needed. Herein, we used an oscillatory shear rheometer to evaluate changes in rheological properties of mouse muscles (tibialis anterior, TA) and their restoration upon autologous cell therapy by comparing the viscoelastic properties of normal, diseased and treated muscles. Amplitude sweep tests of muscle samples were performed under 20% compression over a range of shear strain between 0.01 and 2% and frequency of 1 rad/s. The samples were tested in plane-plane geometry and horizontal myofiber alignment. Typical linear viscoelastic region (LVER) patterns were found for each muscle type. For healthy muscles, a broad LVER between shear deformations (γ) of 0.013–0.62% was observed. The LVER of DMD mdx/SCID muscles was found at 0.14% to 0.46% shear deformation, and no shear dependence of storage (G') and loss (G“) moduli at γ range changing from 0.034% to 0.26% was found for transplanted tissues. G'LVER and G”LVER moduli of healthy muscles were significantly higher than G'LVER and G“LVER of dystrophic tissues. Additionally, muscle resistance assessment by rheometer indicated that muscles transplanted with stem cells restored elastic properties to levels close to those of healthy muscles. Interestingly, histological staining and rheological data indicate that the loss factor is strongly related to structural changes of examined muscles.

Agni is termed as Vaishwanara as it takes the person from Mruthyuloka to Swargaloka. It is an important factor and is equitant to Prana; is one among Dashaprana Ayatana. Agni in Shareera is present in different forms with different actions. It is the responsible factor for both health and disease; on the other hand the successful outcome of treatment is also dependant on Agni. Chikitsa (treatment) is the process of bestowing normalcy which is either brought by Shodhana (purificatory) or Shamana (palliative) Karma. Snehapana is a pre-operative procedure for Shodhana Chikitsa where in Sneha Dravya (medicated fat) is administered for attainment of Upasthita Dosha Avastha and further ease in elimination of the vitiated Doshas. Assessment of Dosha, Dushya, Vyadhi Avastha, Roga Bala, Rogi Bala, Agni, Koshta etc. factors are essential for the attainment of Chikitsa Phala. Assessment of Agni not only helps in understanding Vyadhi but also enables to plan the dosage of Sneha to be administered. Thus this paper is an attempt to throw light on the importance of Agni, assessment of Agni and Agni Bala prior to Shodhananga Snehapana.

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

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.

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. The isolation and identification of active principles and elucidation of the mechanism of action of a drug is of paramount importance. Hence, works in both mixture of traditional medicine and single active compounds are very important. One such compound is metallochlorophyllin, a water soluble analogue of the ubiquitous green pigment chlorophyll. Metallochlorophyllin can neutralize several physically relevant oxidants and may decrease oxidative damage induced by chemical carcinogens and radiation. It also acts as an effective inhibitor of aflatoxin hepatocarcinogenesis in animal models by blocking carcinogen bioavailability. Further anti-cancer effects of metallochlorophyllin including antioxidant activity, inhibition of enzymatic activity that converts inert procarcinogens into active carcinogens; stimulation of enzymatic activity that promotes the elimination of toxic substances from the body; protection from radiation; and antitumor activity have likewise been evidenced by controlled studies. The metallochlorophyllins are compounds worthy of further study, with the potential to provide great benefit to human health.

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.