Bone marrow-derived stromal cells (BMSCs) protect against acute lung injury (ALI). To determine the role of BMSC mitochondria in this protection, we airway-instilled mice first with ...lipopolysaccharide (LPS) and then with either mouse BMSCs (mBMSCs) or human BMSCs (hBMSCs). Live optical studies revealed that the mBMSCs formed connexin 43 (Cx43)-containing gap junctional channels (GJCs) with the alveolar epithelia in these mice, releasing mitochondria-containing microvesicles that the epithelia engulfed. The presence of BMSC-derived mitochondria in the epithelia was evident optically, as well as by the presence of human mitochondrial DNA in mouse lungs instilled with hBMSCs. The mitochondrial transfer resulted in increased alveolar ATP concentrations. LPS-induced ALI, as indicated by alveolar leukocytosis and protein leak, inhibition of surfactant secretion and high mortality, was markedly abrogated by the instillation of wild-type mBMSCs but not of mutant, GJC-incompetent mBMSCs or mBMSCs with dysfunctional mitochondria. This is the first evidence, to our knowledge, that BMSCs protect against ALI by restituting alveolar bioenergetics through Cx43-dependent alveolar attachment and mitochondrial transfer.
Recapitulation of lung development from human pluripotent stem cells (hPSCs) in three dimensions (3D) would allow deeper insight into human development, as well as the development of innovative ...strategies for disease modelling, drug discovery and regenerative medicine. We report here the generation from hPSCs of lung bud organoids (LBOs) that contain mesoderm and pulmonary endoderm and develop into branching airway and early alveolar structures after xenotransplantation and in Matrigel 3D culture. Expression analysis and structural features indicated that the branching structures reached the second trimester of human gestation. Infection in vitro with respiratory syncytial virus, which causes small airway obstruction and bronchiolitis in infants, led to swelling, detachment and shedding of infected cells into the organoid lumens, similar to what has been observed in human lungs. Introduction of mutation in HPS1, which causes an early-onset form of intractable pulmonary fibrosis, led to accumulation of extracellular matrix and mesenchymal cells, suggesting the potential use of this model to recapitulate fibrotic lung disease in vitro. LBOs therefore recapitulate lung development and may provide a useful tool to model lung disease.
The ability to generate lung and airway epithelial cells from human pluripotent stem cells (hPSCs) would have applications in regenerative medicine, modeling of lung disease, drug screening and ...studies of human lung development. We have established, based on developmental paradigms, a highly efficient method for directed differentiation of hPSCs into lung and airway epithelial cells. Long-term differentiation of hPSCs in vivo and in vitro yielded basal, goblet, Clara, ciliated, type I and type II alveolar epithelial cells. The type II alveolar epithelial cells were capable of surfactant protein-B uptake and stimulated surfactant release, providing evidence of specific function. Inhibiting or removing retinoic acid, Wnt and BMP-agonists to signaling pathways critical for early lung development in the mouse-recapitulated defects in corresponding genetic mouse knockouts. As this protocol generates most cell types of the respiratory system, it may be useful for deriving patient-specific therapeutic cells.
The entomopathogenic fungus (EPF), Beauveria bassiana, is reported as the most potent biological control agent against a wide range of insect families. This study aimed to isolate and characterize ...the native B. bassiana from various soil habitats in Bangladesh and to evaluate the bio-efficacy of these isolates against an important vegetable insect pest, Spodoptera litura. Seven isolates from Bangladeshi soils were characterized as B. bassiana using genomic analysis. Among the isolates, TGS2.3 showed the highest mortality rate (82%) against the 2nd instar larvae of S. litura at 7 days after treatment (DAT). This isolate was further bioassayed against different stages of S. litura and found that TGS2.3 induced 81, 57, 94, 84, 75, 65, and 57% overall mortality at egg, neonatal 1st, 2nd, 3rd, 4th, and 5th instar larvae, respectively, over 7 DAT. Interestingly, treatment with B. bassiana isolate TGS2.3 resulted in pupal and adult deformities as well as decreased adult emergence of S. litura. Taken together, our results suggest that a native isolate of B. bassiana TGS2.3 is a potential biocontrol agent against the destructive insect pest S. litura. However, further studies are needed to evaluate the bio-efficacy of this promising native isolate in planta and field conditions.
Lung fibrosis is increasingly detected with aging and has been associated with poor outcomes in acute lung injury or infection. However, the molecular programs driving this pro-fibrotic evolution are ...unclear. Here we profile distal lung samples from healthy human donors across the lifespan. Gene expression profiling by bulk RNAseq reveals both increasing cellular senescence and pro-fibrotic pathway activation with age. Quantitation of telomere length shows progressive shortening with age, which is associated with DNA damage foci and cellular senescence. Cell type deconvolution analysis of the RNAseq data indicates a progressive loss of lung epithelial cells and an increasing proportion of fibroblasts with age. Consistent with this pro-fibrotic profile, second harmonic imaging of aged lungs demonstrates increased density of interstitial collagen as well as decreased alveolar expansion and surfactant secretion. In this work, we reveal the transcriptional and structural features of fibrosis and associated functional impairment in normal lung aging.
Acute Lung Injury (ALI) due to inhaled pathogens causes high mortality. Underlying mechanisms are inadequately understood. Here, by optical imaging of live mouse lungs we show that a key mechanism is ...the viability of cytosolic Ca
buffering by the mitochondrial Ca
uniporter (MCU) in the lung's surfactant-secreting, alveolar type 2 cells (AT2). The buffering increased mitochondrial Ca
and induced surfactant secretion in wild-type mice, but not in mice with AT2-specific MCU knockout. In the knockout mice, ALI due to intranasal LPS instillation caused severe pulmonary edema and mortality, which were mitigated by surfactant replenishment prior to LPS instillation, indicating surfactant's protective effect against alveolar edema. In wild-type mice, intranasal LPS, or Pseudomonas aeruginosa decreased AT2 MCU. Loss of MCU abrogated buffering. The resulting mortality was reduced by spontaneous recovery of MCU expression, or by MCU replenishment. Enhancement of AT2 mitochondrial buffering, hence endogenous surfactant secretion, through MCU replenishment might be a therapy against ALI.
Plant growth promoting rhizobacteria (PGPR) have been intensively investigated in agricultural crops for decades. Nevertheless, little information is available on the application of Sphingomonas spp. ...as a PGPR particularly in vegetables, despite of potential plant growth promoting traits of this group. This study investigated the role of Sphingomonas panaciterrae (PB20) on growth and nutritional profile of spinach applied through seed priming (SP), soil drenching (SD), foliar application (FA), and bacterial culture filtrate foliar (BCF) applications. The results showed that, depending on different methods of application, PB20 significantly increased plant height (19.57–65.65 %), fresh weight (7.26–37.41 %), total chlorophyll (71.14–192.54 %), carotenoid (67.10–211.67 %) antioxidant (55.99–207.04), vitamin C (8.1–94.6 %) and protein content (6.7–21.5 %) compared to control in the edible part of spinach. Among the mineral nutrients, root nitrogen (N) showed greater response to bacterial application (18.65%–46.15 % increase over control) than shoot nitrogen (6.70%–21.52 % increased over control). Likewise, in all methods of application, phosphorus (P) content showed significant increase over control both in root (42.79–78.48 %) and in shoot (3.57–27.0 %). Seed priming and foliar application of PB20 increased the shoot calcium (Ca) content compared to control. BCF foliar application yielded maximum magnesium (Mg), iron (Fe) and zinc (Zn) in shoot. However, seed priming resulted in maximum Fe in root. Overall, seed priming outperformed in growth, vitamin C, antioxidants, N and P uptake, while BCF foliar application resulted in better uptake of several nutrients. Multivariate analysis validated the positive association of most of the growth parameters with SP while several nutrients with FA and BCF. Based on the findings it is evident that this rhizobacteria PB20 has the potentiality to be applied as a biofertilizer to produce nutrient-enriched spinach with an improved yield. Farmers can conveniently incorporate PR20 through seed priming before planting of spinach, with additional benefits through foliar spray.
•PGPR Sphingomonas panaciterrae PB20 applied in spinach by four different methods.•PB20 improved plant height, fresh weight, antioxidant, vitamin C, chlorophyll, and carotenoid content of spinach plant.•PB20 increased the amount of N, P, Ca and Mg, Fe and Zn in the shoot of spinach in all methods of application.•Seed priming resulted the tallest plant, highest amount of antioxidant and N and P content in spinach shoot and root.•Foliar application and bacterial culture filtrate foliar application showed better results in several nutrients uptake.
The potentiality of barnyard grass for remediation of arsenic (As)-contaminated soil has been reported in several research works. However, the phytoremediation ability of barnyard grass from ...industrially polluted multimetal-contaminated soil in comparison to As-amended soil needs to be elucidated. This work investigated the As remediation potentiality of barnyard grass from As-amended and industrially polluted soils, and the fractionation of As was done in soils with plants and without plants grown. The result showed that at the highest As level in the soil, barnyard grass accumulated the highest amount of As in both the root (414.81 mg kg
-1
) and shoot (114.12 mg kg
-1
). However, barnyard grass produced the highest amount of biomass in industrially polluted soil that resulted in the highest amount of As uptake. Moreover, barnyard grass also accumulated lead (Pb) and chromium (Cr) from industrially polluted soil. The bioaccumulation factor (BF) of As was >1 in As-amended soil in all the treatments as well as in industrially polluted soil. Fractionation of As in post-harvest soil revealed that compared to soil without plants grown, As in the soil was reduced from residual As (F5); As associated with well-crystallized hydrous oxides of iron (Fe) and aluminum (Al) (F4); As associated with amorphous and poorly crystallized hydrous oxides of Fe and Al (F3), whereas a slight increase was found in non-specifically sorbed As (F1) and specifically sorbed As (F2) due to the plant’s effect. The slight increase in the concentration of As in F1 and F2 fractions contributed to the bioavailable forms of As in the rhizosphere and sustained As concentration for further plant uptake. The maximum plant growth and highest uptake of As in the industrially polluted soil revealed the potentiality of barnyard grass for remediation of multimetal-polluted soil.
Rhizoctonia solani is an important necrotrophic pathogenic fungus that causes okra root disease and results in severe yield reduction. Many biocontrol agents are being studied with the intent of ...improving plant growth and defense systems and reducing crop loss by preventing fungal infections. Recently, a member of the Hypocrealean family, Metarhizium anisopliae, has been reported for insect pathogenicity, endophytism, plant growth promotion, and antifungal potentialities. This research investigated the role of M. anisopliae (MetA1) in growth promotion and root disease suppression in okra. The antagonism against R. solani and the plant growth promotion traits of MetA1 were tested in vitro. The effects of endophytic MetA1 on promoting plant growth and disease suppression were assessed in planta. Dual culture and cell-free culture filtrate assays showed antagonistic activity against R. solani by MetA1. Some plant growth promotion traits, such as phosphate solubilization and catalase activity were also exhibited by MetA1. Seed primed with MetA1 increased the shoot, root, leaves, chlorophyll content, and biomass content compared to control okra plants. The plants challenged with R. solani showed the highest hydrogen peroxide (H2O2) and lipid peroxidation (MDA) contents in the leaves of okra. Whereas MetA1 applied plants showed a reduction of H2O2 and MDA by 5.21 and 14.96%, respectively, under pathogen-inoculated conditions by increasing antioxidant enzyme activities, including catalase (CAT), peroxidase (POD), glutathione S-transferase (GST), and ascorbate peroxidase (APX), by 30.11, 10.19, 5.62, and 5.06%, respectively. Moreover, MetA1 increased soluble sugars, carbohydrates, proline, and secondary metabolites, viz., phenol and flavonoid contents in okra resulting in a better osmotic adjustment of diseases infecting plants. MetA1 reduced disease incidence by 58.33% at 15 DAI compared to the R. solani inoculated plant. The results revealed that MetA1 improved plant growth, elevated the plant defense system, and suppressed root diseases caused by R. solani. Thus, MetA1 was found to be an effective candidate for the biological control program.
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