Despite the phytotoxicity of olive-mill solid waste (OMSW) due to its high polyphenols content, OMSW have fertilizer characteristics, which make it a potential source for organic fertilization. Composting of OMSW treatment process was conducted in this study to eliminate the phytotoxicity and solve the environmental impact of this waste. Recycling of OMSW was carried out via composting of six batches of trials using equal proportions of OMSW, cow manure (C) and wheat straw (W). The treatment process was performed at two time intervals (two and five months), after each one, the recipient species (Vicia faba L.) was planted. The results showed the efficiency of composting in reducing OMSW original toxicity after two months than five months. The germination percentage and the plumule and radicle lengths of V. faba showed a significant improvement when the OMSW was composted with C at different proportions before using as soil amendments. Besides, the total biomass was noticeably increased at the high concentration of C-OMSW. Similarly, the total pigments concentration in V. faba was increased by using various composts after two months, where the highest pigment content was observed at 40% W-OMSW treatment.
We investigated whether particles suitable for delivery to alveolar macrophages may provide a means of targeting rapamycin, an inducer of autophagy, to alveolar macrophages as a host-directed antituberculosis agent. Inhalable particles were prepared by spray-drying and characterized using laser scattering and electron microscopy. Their aerodynamic diameter was calculated from bulk and tapped densities. In vitro drug release was studied in PBS containing 1% SDS. In vitro uptake of particles by THP-1 derived macrophages was studied by flow cytometry. Cytotoxicity of the particles toward macrophages and their efficacy against intracellular Mycobacterium tuberculosis were studied using a methyltetrazolium assay and counting bacterial colonies obtained when cell lysates were plated on agar. The encapsulation efficiency was 88.8 ± 1.13% and drug content 22 ± 4% w/w. The particles had a median diameter of 2.88 ± 0.8 μm and appeared as collapsed spheres. Their calculated aerodynamic diameter was about 1 μm. In vitro drug release from the particles was first-order and extended beyond 10 days. Flow cytometry indicated that the particles were taken up by macrophages within 3 h. Macrophages exposed to the particles or rapamycin in solution at a concentration of 100 μg/mL over a 24 h period maintained 79.37 ± 0.72% and 58.33 ± 1.39% viability, respectively. Efficacy studies concluded that particles were more effective in clearing intracellular mycobacteria than rapamycin in solution. It was concluded that the preparation was suitable for formulating as a dry powder inhalation to test efficacy of inhaled, macrophage-targeted rapamycin against TB.
Despite the phytotoxicity of olive-mill solid waste (OMSW) due to its high polyphenols content, OMSW have fertilizer characteristics, which make it a potential source for organic fertilization. Composting of OMSW treatment process was conducted in this study to eliminate the phytotoxicity and solve the environmental impact of this waste. Recycling of OMSW was carried out via composting of six batches of trials using equal proportions of OMSW, cow manure (C) and wheat straw (W). The treatment process was performed at two time intervals (two and five months), after each one, the recipient species (Vicia faba L.) was planted. The results showed the efficiency of composting in reducing OMSW original toxicity after two months than five months. The germination percentage and the plumule and radicle lengths of V. faba showed a significant improvement when the OMSW was composted with C at different proportions before using as soil amendments. Besides, the total biomass was noticeably increased at the high concentration of C-OMSW. Similarly, the total pigments concentration in V. faba was increased by using various composts after two months, where the highest pigment content was observed at 40% W-OMSW treatment.The maximum uptake of potassium and sodium was recorded through the application of W-OMSW compost to soil after two months. Furthermore, the C-OMSW composts showed the highest concentration of nitrogen, calcium, iron and manganese. However, the C-W-OMSW composts recorded the highest concentration of phosphorous, magnesium and copper. Finally, this study developed a low cost treatment that will enable the growers to convert OMSW into a natural nontoxic compost rich with essential nutrients which have positive effects on plants growth.
PE11 (Rv1169c or LipX) is a cell wall associated esterase/lipase of Mycobacterium tuberculosis (Mtb). Evidences suggest that PE11 is expressed by Mtb both in vitro and in vivo. Previous studies have shown that PE11 leads to modification in cell wall lipid content and enhanced virulence when expressed in the non-pathogenic surrogate Mycobacterium smegmatis. Since cell wall lipids often play different roles in pathogenic and non-pathogenic mycobacteria, we investigated the role of PE11 in its host, Mtb. Mtb with lowered expression of PE11 (PE11 knock-down) displayed significant changes in colony morphology and cell wall lipid profile, confirming the role of PE11 in cell wall architecture. In addition, the levels of phthiocerol dimycocerosates, a cell wall virulence factor, were decreased. Levels of trehalose esters and free mycolic acids were increased. In contrast to M. smegmatis expressing Mtb PE11, a role reversal was observed in Mtb with respect to pellicle/biofilm formation. The PE11 knock-down Mtb strain showed significantly enhanced aggregation and early biofilm growth in detergent-free medium, compared to the wild-type. Knock-down strain also showed nearly 27-fold up-regulation of a fibronectin attachment protein (Rv1759c), linking biofilm growth with over-expression of bacterial proteins that help in aggregation and/or binding to host extracellular matrix. The knock-down also resulted in poor virulence of Mtb in PMA (phorbol 12-myristate 13-acetate) treated and PMA+IFN-γ treated THP-1 macrophages. Therefore, the study not only links PE11 to cell wall virulence lipids but also reveals the involvement of this cell wall associated esterase in down-regulation of biofilm in Mtb.
Cytotoxic frog antimicrobial peptide Temporin L (TempL) is an attractive molecule for the design of lead antimicrobial agents due to its short size and versatile biological activities. However, noncytotoxic TempL variants with desirable biological activities have rarely been reported. TempL analogue Q3K,TempL is water-soluble and possesses a significant antiendotoxin property along with comparable cytotoxicity to TempL. A phenylalanine residue, located at the hydrophobic face of Q3K,TempL and the “d” position of its phenylalanine zipper sequence, was replaced with a cationic lysine residue. This analogue, Q3K,F8K,TempL, showed reduced hydrophobic moment and was noncytotoxic with lower antimicrobial activity. Interestingly, swapping between tryptophan at the fourth and serine at the sixth positions turned Q3K,F8K,TempL totally amphipathic as reflected by its helical wheel projection with clusters of hydrophobic and hydrophilic residues and the highest hydrophobic moment among these peptides. Surprisingly, this analogue, SW,Q3K,F8K,TempL, was as noncytotoxic as Q3K,F8K,TempL but showed augmented antimicrobial and antiendotoxin properties, comparable to that of TempL and Q3K,TempL. SW,Q3K,F8K,TempL exhibited appreciable survival of mice against P. aeruginosa infection and a lipopolysaccharide (LPS) challenge. Unlike TempL and Q3K,TempL, SW,Q3K,F8K,TempL adopted an unordered secondary structure in bacterial membrane mimetic lipid vesicles and did not permeabilize them or depolarize the bacterial membrane. Overall, the results demonstrate the design of a nontoxic TempL analogue that possesses clusters of hydrophobic and hydrophilic residues with impaired secondary structure and shows a nonmembrane-lytic mechanism and in vivo antiendotoxin and antimicrobial activities. This paradigm of design of antimicrobial peptide with clusters of hydrophobic and hydrophilic residues and high hydrophobic moment but low secondary structure could be attempted further.
Naphthoquinones are known to exhibit a broad range of biological activities against microbes, cancer and parasitic diseases and have been widely used in Indian traditional medicine. Plumbagin is a plant-derived naphthoquinone metabolite (5-hydroxy-2-methyl-1,4-naphthoquinone) reported to inhibit trypanothione reductase, the principal enzyme and a validated drug target involved in detoxification of oxidative stress in Leishmania. Here, we report the mechanistic aspects of cell death induced by plumbagin including physiological effects in the promastigote form and ultrastructural alterations in both promastigote and amastigote forms of Leishmania donovani which till now remained largely unknown. Our observations show that oxidative stress induced by plumbagin resulted in depolarization of the mitochondrial membrane, depletion in ATP levels, elevation of cytosolic calcium, increase in caspase 3/7-like protease activity and lipid peroxidation in promastigotes. Apoptosis-like cell death induction post plumbagin treatment was confirmed by biochemical assays like Annexin V/FITC staining, TUNEL as well as morphological and ultrastructural studies. These findings collectively highlight the mode of action and importance of oxidative stress inducing agents in effectively killing both forms of the Leishmania parasite and opens up the possibility of exploring plumbagin and its derivatives as promising candidates in the chemotherapy of Leishmaniasis.
Indole acetic acid (IAA) production is a major property of rhizosphere bacteria that plays a central role in plant growth and development as a regulator of cell division, cell differentiation, cell expansion, lateral root formation, flowering, and tropic responses. The present study deals with isolation, functional characterization and identification of IAA producing rhizobacteria from the rhizosphere. The present study deals with the emerging need to protect medicinal plants that represent our natural resources. A total ten bacterial isolates were recovered from rhizospheric soil associated with Withania somnifera, recognized as bacillus spp. by morphotpic and biochemical characterization and tested for indole acetic acid production. Out of ten rhizobacterial isolates, six were selected as efficient Producers of IAA. The amount of indole acetic acid produced was detected in the presence of different concentration of tryptophan, which lead to the elevation in indole acetic acid production as compared in the absence of tryptophan. The IAA production was further confirmed by thin layer chromatography (TLC). The impact of PGPR was evaluated on Withania somnifera seeds that showed significant elevation in germination %, root and shoot length as compared to the untreated seeds. Out of the chosen PGPR, WSNb3, WSNb5 and WSNb6 were showed to be the best to produce IAA. Subsequently, effect on plant growth was tested by towel paper assay. In conclusion the study suggests the IAA producing bacteria may be efficient biofertilizer inoculants to promote plant growth and protecting the medicinal plants for the future generation.
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