Membrane Disintegration Research Articles

On the rapid in situ oxidation of two-dimensional V2CTz MXene in culture cell media and their cytotoxicity

The plethora of emerging two-dimensional (2D) materials exhibit wide potential application in novel technologies and advanced devices. However, their stability in environmental conditions could be an issue, affecting their application possibilities and posing health risks. Moreover, their decomposed leftovers can also induce a negative influence on human health. In particular, transition metal carbides commonly referred to as MXenes are susceptible to environmental oxidation being decomposed toward transition metal oxides and carbide-derived carbon. In this study we focused on the oxidation-state-related in vitro cytotoxicity of delaminated V2CTz onto immortalized keratinocytes (HaCaT) and malignant melanoma (A375) human cell lines. Due to the fact, that the V2CTx MXenes are least stable from all known obtained MXenes up to date, the vanadium ones were a practical choice to visualize the oxidation-cytotoxic correlation keeping the standards of 24–48 h of cell culturing. We found that the oxidation of V2CTz highly increases their cytotoxicity toward human cells, which is also time and dose dependent. The identified mode of action relates to the cell cycle as well as cellular membrane disintegration through direct physicochemical interactions.

The Penicillium chrysogenum Q176 Antimicrobial Protein PAFC Effectively Inhibits the Growth of the Opportunistic Human Pathogen Candida albicans.

Small, cysteine-rich and cationic antimicrobial proteins (AMPs) from filamentous ascomycetes promise treatment alternatives to licensed antifungal drugs. In this study, we characterized the Penicillium chrysogenum Q176 antifungal protein C (PAFC), which is phylogenetically distinct to the other two Penicillium antifungal proteins, PAF and PAFB, that are expressed by this biotechnologically important ascomycete. PAFC is secreted into the culture broth and is co-expressed with PAF and PAFB in the exudates of surface cultures. This observation is in line with the suggested role of AMPs in the adaptive response of the host to endogenous and/or environmental stimuli. The in silico structural model predicted five β-strands stabilized by four intramolecular disulfide bonds in PAFC. The functional characterization of recombinant PAFC provided evidence for a promising new molecule in anti-Candida therapy. The thermotolerant PAFC killed planktonic cells and reduced the metabolic activity of sessile cells in pre-established biofilms of two Candida albicans strains, one of which was a fluconazole-resistant clinical isolate showing higher PAFC sensitivity than the fluconazole-sensitive strain. Candidacidal activity was linked to severe cell morphology changes, PAFC internalization, induction of intracellular reactive oxygen species and plasma membrane disintegration. The lack of hemolytic activity further corroborates the potential applicability of PAFC in clinical therapy.

Discovery and mechanistic studies of cytotoxic cyclotides from the medicinal herb Hybanthus enneaspermus

Cyclotides are plant-derived peptides characterized by an ∼30-amino acid-long cyclic backbone and a cystine knot motif. Cyclotides have diverse bioactivities, and their cytotoxicity has attracted significant attention for its potential anticancer applications. Hybanthus enneaspermus (Linn) F. Muell is a medicinal herb widely used in India as a libido enhancer, and a previous study has reported that it may contain cyclotides. In the current study, we isolated 11 novel cyclotides and 1 known cyclotide (cycloviolacin O2) from H. enneaspermus and used tandem MS to determine their amino acid sequences. We found that among these cyclotides, hyen C comprises a unique sequence in loops 1, 2, 3, 4, and 6 compared with known cyclotides. The most abundant cyclotide in this plant, hyen D, had anticancer activity comparable to that of cycloviolacin O2, one of the most cytotoxic known cyclotides. We also provide mechanistic insights into how these novel cyclotides interact with and permeabilize cell membranes. Results from surface plasmon resonance experiments revealed that hyen D, E, L, and M and cycloviolacin O2 preferentially interact with model lipid membranes that contain phospholipids with phosphatidyl-ethanolamine headgroups. The results of a lactate dehydrogenase assay indicated that exposure to these cyclotides compromises cell membrane integrity. Using live-cell imaging, we show that hyen D induces rapid membrane blebbing and cell necrosis. Cyclotide-membrane interactions correlated with the observed cytotoxicity, suggesting that membrane permeabilization and disintegration underpin cyclotide cytotoxicity. These findings broaden our knowledge on the indigenous Indian herb H. enneaspermus and have uncovered cyclotides with potential anticancer activity.

Effects of pulsed electric field pretreatment on frying quality of fresh-cut lotus root slices

This study aimed to compare the effects of pulsed electric field (PEF) pretreatment on fresh-cut lotus root and its frying quality with those under blanching (80 °C, 3 min). The lotus root slices were treated by 10,000 pulses at different electric field strength (0.5, 1.0 and 1.5 kV/cm) with specific energy levels (5.56, 22.20 and 50.00 kJ/kg). PEF treatment decreased the reducing sugar content in fresh samples and then diminished the Millard reaction, including lower browning index and 25.9% reduction in acrylamide content. Besides, membrane disintegration occurred when the cells were subjected to PEF, resulting in soft tissue structure and a reduction of oil inhalation of 15%. Results imply that PEF treatment can be used as a viable alternative to blanching to pretreat lotus root slices, effectively saved thermal energy required for industrial production and delivered more positive effects on quality of fried slices.

Simultaneous Ultrasensitive Detection and Elimination of Drug-Resistant Bacteria by Cyclometalated Iridium(III) Complexes.

Antimicrobial resistance has become a major threat to public health due to the rampant and empirical use of antibiotics. Rapid diagnosis of bacteria with the desired sensitivity and selectivity still, however, remains an open challenge. We report a special class of water-soluble metal-based aggregation-induced emission luminogens (AIEgens), namely, cyclometalated iridium(III) polypyridine complexes of the type [Ir(PQ)2(N^N)]Cl (1-3), where PQ = 2-phenylquinoline and N^N = 2,2'-bipyridine derivatives, that demonstrate dual capability for detection and elimination of drug-resistant bacteria in aqueous solutions. These AIEgens exhibit selective and rapid sensing of endotoxins, such as lipopolysaccharides (LPS) and lipoteichoic acid (LTA) released by the bacteria, with a detection limit in the lower nanomolar range. Targeting these naturally amplified biomarkers (approximately 1 million copies per cell) by iridium(III) complexes induces strong AIE in the presence of different Gram-negative and Gram-positive bacteria including carbapenem-resistant A. baumannii (CRAB) and methicillin-resistant S. aureus (MRSA) at concentrations as low as 1.2 CFU/mL within 5 min in spiked water samples. Detection of bacteria by the complexes is also visible to the naked eye at higher (108 CFU/mL) cell concentrations. More notably, complexes 1 and 2 show potent antibacterial activity against drug-resistant bacteria with low minimum inhibitory concentrations (MICs) ≤ 5 μg/mL (1-4 μM) via ROS generation and cell membrane disintegrity. To the best of our knowledge, this work is the "first-in-class" example of a metal-based theranostic system that integrates selective, sensitive, rapid, naked-eye, wash-free, and real-time detection of bacteria using broad-spectrum antibiotics into a single platform. This dual capability of AIEgens makes them ideal scaffolds for monitoring bacterial contamination in aqueous samples and pharmaceutical applications.

Superabsorbent sodium carboxymethyl cellulose membranes based on a new cross-linker combination for female sanitary napkin applications

The main intent of this investigation was to retain the strength and superabsorbency of natural and non-toxic biodegradable polymers using an innovative combination of cross-linkers for application as the absorbent core of sanitary napkins. For this, sodium carboxymethyl cellulose (NaCMC) and starch were blend to form membranes by phase inversion and lyophilisation, using an optimized cross-linker combination of sodium trimetaphosphate (STMP) and aluminium sulphate (AlS). Optimal cross-linking of NaCMC and starch hampered membrane dissolution and disintegration, yielding a microtextured surface morphology. The membranes were biodegradable and yet possessed the requisite flexibility and mechanical strength for the proposed application, without compromise of superabsorbency. Lyophilised membranes possessed higher immediate water and blood sorption with ∼50% water retention capabilities when compared to the phase inversion technology. The results suggest that the developed membranes can be a cost-effective degradable alternative to the commercial polyacrylate-based nonbiodegradable sanitary products.

Highlighting the effect of amyloid beta assemblies on the mechanical properties and conformational stability of cell membrane

Alzheimer disease (AD) is the most common cause of dementia, characterized by a progressive decline in cognitive function due to the abnormal aggregation and deposition of Amyloid beta (Aβ) fibrils in the brain of patients. In this context, the molecular mechanisms of protein misfolding and aggregation that are known to induce significant biophysical alterations in cells, including destabilization of plasma membranes, remain partially unclear. Physical interaction between the Aβ assemblies and the membrane leads to the disruption of the cell membrane in multiple ways including, surface carpeting, generation of transmembrane channels and detergent-like membrane dissolution. Understanding the impact of amyloidogenic protein in different stages of aggregation with the plasma membrane, plays a crucial role to fully elucidate the pathological mechanisms of AD. Within this framework, computer simulations represent a powerful tool able to shed lights on the interactions governing the structural influence of Aβ proteins on biological membrane. In this study, molecular dynamics (MD) simulations have been performed in order to characterize how POPC bilayer conformational and mechanical properties are affected by the interaction with Aβ11-42 peptide, oligomer and fibril.

Anticancer pentamethinium salt is a potent photosensitizer inducing mitochondrial disintegration and apoptosis upon red light illumination

Despite progress in the development and application of novel therapeutic agents, cancer remains a major cause of death worldwide. Therefore, there is a need for new approaches to increase therapeutic options and efficiency. The metabolism of cancer cells differs from that of non-malignant cells and their mitochondria show altered activities that can be utilized as a target for drug development. Salt 1 is a low-molecular weight heterocyclic compound of the polymethine class that accumulates in the mitochondria of cancer cells and selectively disrupts their metabolism. Salt 1 leads to a non-apoptotic form of cell death in vitro that is associated with an autophagic cellular response and eventual metabolic collapse, and inhibits human tumor xenograft growth in vivo without apparent toxicity for normal cells. As a pentamethinium compound, salt 1 exhibits intrinsic fluorescence and is a candidate for photosensitization after excitation by appropriate wavelengths of light. Herein, we report that salt 1 is a potent photosensitizer, which generates a photodynamic effect and provides enhanced cytotoxicity compared to salt 1 without light exposure. Importantly, photosensitization is optimally induced by red light, which is used clinically for photosensitization and penetrates further into tissues than lower wavelengths. Cancer cells treated with non-cytotoxic doses of salt 1 and subsequently exposed to 630 nm light show severely damaged mitochondria, manifested by reduced mitochondrial membrane potential and disintegration of the mitochondrial tubular network. As a consequence, cancer cells lose their proliferative potential and die via apoptosis in the presence of light. These findings indicate that salt 1 is a promising photosensitizer with potential to be combined with 630 nm light to strengthen its efficacy in cancer therapy.

Toxicity of pesticides toward human immune cells U-937 and HL-60

The industrialization of the agricultural sector has significantly increased the use of chemicals such as pesticides. Therefore, exposure to them is unavoidable, which makes it necessary to assess their safety for humans at actual exposure doses. This paper aims to determine toxicity of three types of pesticides toward human immune cells (HL-60 and U-937): glyphosate (GLY), deltamethrin (DEL), and chlorothalonil (CHL). Cell viability, membrane integrity, inflammation induction, and antioxidant activity were evaluated to determine differences in cellular response to the tested plant protection agents. In experimental models, all tested substances caused increased mortality of cells after only 24 h. Cell membrane damage was recorded under DEL and CHL influences. The largest disintegration of the cell membrane was due to the action of 100 μg/mL DEL for U-937 and CHL at 1 μg/mL for HL-60. GLY at a concentration of 3,600 μg/mL caused significant peroxidation of U-937 cells’ lipids. CHL-induced inflammation in both types of cells tested. DEL and GLY also induced antioxidant activity in cells. These results lead to the conclusion that the tested pesticides act cytotoxically to the cells of the human immune system in doses to which both farmers and consumers are exposed.

Microvesicles derived from squamous cell carcinoma induce cell death, autophagy, and invasion of benign myoepithelial cells.

There has been great interest recently in the mechanisms of cell-to-cell communication through microvesicles (MV). These structures are produced by many different cell types and can modulate cellular activity by induction of epigenetic alterations. These vesicles may promote tumor mass increase either by stimulating cell proliferation via growth factors or by inhibiting apoptosis, which reinforces the role of such vesicles as important modulators of tumor progression. The present in vitro study aimed to characterize MV derived from malignant neoplastic epithelial cell cultures (EP) and their effect on the expression of apoptosis/autophagy and invasion related genes of benign myoepithelial (Myo) cell cultures. The results revealed round structures with a mean size of 153.6 (±0.2) nm, with typical MV morphology. CD63 quantification indicated that EP cell culture at 70%-80% confluence secreted 3.088×108 MV/mL. Overall, Myo exposed to MVs derived from EP showed both up- and downregulation of tumorigenesis promoting genes. MVs from EP cells promoted cell death of Myo cells and positively modulate BAX, SURVIVIN, LC3B, MMP-2, and MMP-9 expression. Furthermore, an increasing of MMP-2 and MMP-9 secretion by Myo was observed after MV exposure. These findings suggest that MVs from EP modulate autophagy of Myo cells, which may, in part, explain the disappearance of these cells in in situ areas of invasive carcinoma ex-pleomorphic adenoma. Additionally, the overexpression of MMPs contributes to the development of an invasive phenotype of Myo cells, which could favor the dissolution of the basement membrane during tumorigenesis process.

Bisphenol-A exposure induced neurotoxicity and associated with synapse and cytoskeleton in Neuro-2a cells

Bisphenol A (BPA) is an environmental chemical that induces neurotoxic effects for human. Synaptophysin (SYP) and drebrin (Dbn) proteins are involved in regulating synaptic morphology. The stability of the cytoskeleton in nerve cells in the brain is regulated by Tau and MAP2. This study aimed to determine the toxicity of BPA to Neuro-2a cells by investigating the synaptic and cytoskeletal damage induced in these cells by 24 h of exposure to 0 (MEM), 50, 100, 150, or 200 μM BPA or DMSO. MTT and LDH assays showed that the death rates of Neuro-2a cells increased, as the BPA concentration increased. Ultrastructural assays revealed that cells underwent nucleolar swelling as well as nuclear membrane and partial mitochondrial dissolution or condensation, following BPA exposure. Morphological analysis further revealed that compared with the cells in the control group, the cells in the BPA-treated groups shrank, became rounded, and exhibited a reduced number of synapses. BPA also significantly decreased the relative protein and mRNA expression levels of Dbn, MAP2 and Tau (P < .01), but increased the relative protein and mRNA expression levels of SYP (P < .01). These results indicated that BPA suppressed the development and proliferation of Neuro-2a cells by disrupting cellular and synaptic integrity and inflicting cytoskeleton injury.

The outcome of potassium dichromate exposure on histological structure of male rat sexual organs in suckling period

The aim of this study was the evaluation of integrity biomarkers of reproductive toxicity: histoarchitecture of genital organs (testis and epididymis) and sexual accessory glands (prostate, seminal vesicles and bulbo-urethral glands) at sexual maturity after exposure to potassium dichromate (Cr VI) in suckling period. Consequent to the exposure of male rat pups to hexavalent chromium during suckling period structural changes appeared in genital organs and sexual accessory glands, such as: basal membrane and epithelial disintegration, interstitial edema, wavy basal membrane due to a reduction in tubule diameter, Leydig cell necrosis; epithelial smoothing, epithelial necrosis; epithelial cells desquamation and falling in lumen. Exposure to potassium dichromate (Cr VI) during suckling period produced severe congestive and degenerative lesions in genital organs and sexual accessory glands.

Aprepitant Sensitizes Acute Myeloid Leukemia Cells to the Cytotoxic Effects of Cytosine Arabinoside in vitro and in vivo.

PurposeAcute myeloid leukemia (AML) is a complex malignancy characterized by the clonal expansion of immature myeloid precursors. The standard treatment for newly diagnosed AML is chemotherapy consisting of cytosine arabinoside (Ara-C) and anthracyclines with disappointing clinical outcomes and severe adverse effects, such as symptomatic bradycardia, neurotoxicity. Thus, it is promising to treat AML through combination drug therapy to reduce the adverse effects of chemotherapeutics. In our recent published PNAS paper, we reported that NK-1R antagonists, both Aprepitant and SR140333, induce apoptosis of myeloid leukemia cells by inducing oxidative stress through mitochondrial calcium overload. We, therefore, tested the hypothesis of the combination Ara-C with NK-1R antagonist could enhance the efficacy of Ara-C.MethodsMTT assay was employed to detect the cell proliferation. Flow cytometry was applied to detect the cell cycle and necrosis. PI uptake and LDH release assay were used to detect the disintegration of the plasma membrane. Xenograft model was constructed to explore the effect of combination Ara-C with Aprepitant in vivo.ResultsOur results showed that Aprepitant sensitizes HL60 cells to the cytotoxic effects of Ara-C more than 5-fold by enhancing G0/G1 cell cycle arrest and necrosis in vitro. Furthermore, Nec-1, a specific inhibitor of necroptosis, could recover the cell proliferative viability significantly. Attractively, once every 2-days regimen of Ara-C (5 mg/kg) and Aprepitant (10 mg/kg) via in situ injection dramatically reduced the tumor volume from 2175.0 ± 341.9 mm3 in the vehicle group to 828.4 ± 232.4 mm3 in the combination group without obvious toxicity in human myeloid leukemia xenograft mice.ConclusionTaken together, reduced dose of Ara-C combination with moderate Aprepitant provides more effective therapeutical methods for AML treatment in vitro and in vivo with the elimination of the toxicity of Ara-C, which may pay new avenue for the usage of the routine chemotherapy drug Ara-C with low dose to enhance efficacy and reduce toxicity in clinical practice.

Microanalysis of Fish Ovarian Follicular Atresia: A Possible Synergic Action of Somatic and Immune Cells.

The present study describes in detail the morphological characteristics of the process of ovarian follicular atresia in Redbelly tilapia (Coptodon zillii) during the nonbreeding season using light and electron microscopy and immunohistochemistry. The follicular regression process was initiated with shrinkage and disintegration of the nuclear membrane of oocytes resulting in dispersing of chromatin within the ooplasm, followed by marked hyperplasia and hypertrophy of follicular and granulosa cells, which exhibited a strong phagocytic activity to engulf the liquefied yolk particles. Rodlet cells and granulocytes were recorded on the follicular wall and invaded the regressed follicles. Rodlet cells expressed a strong immunoreactivity to matrix metalloperoxidase (MMP-9) and α-smooth muscle actin, while neutrophils expressed a strong reactivity to Myeloperoxidase-3 (MPO). In the advanced stage of follicular atresia, the yolk was almost phagocytized and resorbed and the regressed follicle lost its integrity and appeared to be formed of a cellular mass of phagocytic cells. Transmission electron microscopy revealed the presence of neutrophils, eosinophils, and dendritic cells within the atretic follicle in between these phagocytic cells. Moreover, numerous lysosomes, granules, and phagosomes were observed within the cytoplasm of both phagocytic cells and granulocytes. Telocytes were also demonstrated within the highly thickened richly vascularized theca layer during the late stages of follicular atresia. Immunohistochemical staining for caspase-3 established the participation of apoptosis in the advanced stages of follicular regression. Immune cells, rodlet cells, and telocytes in combination with follicular cells play an essential role in follicular atresia. In conclusion, the present study provides a new evidence on the role of both somatic and immune cells in the phenomenon of ovarian follicular atresia in Redbelly tilapia (Coptodon zillii) during the nonbreeding season.

High Performance and Durable Membrane Electrode Assemblies with Binder-Free Plasma Sprayed Electrodes and Nanocomposite Membranes for Anion Exchange Membrane Electrolyzer

Hydrogen production via water electrolysis using electricity from renewable power sources is a promising method for storing energy in a large scale. Anion exchange membrane (AEM) electrolyzers have the potential for decreasing the hydrogen cost produced by renewables due to use of non-noble materials and inexpensive components for the membrane electrode assembly (MEA). This work demonstrates a highly active and durable MEA for AEM electrolysis using binder-free atmospheric plasma sprayed (APS) RANEY®-type nickel and RANEY®-type nickel-molybdenum electrodes,[1,2] as anode and cathode, respectively in combination with robust two-dimensional hexagonal boron nitride (h-BN)-based anion exchange membranes, nanocomposed with either polyvinylidene fluoride (PVDF) or polyvinyl alcohol (PVA). In this present work, various AEMs with different ionic conductivity and thickness, M1: High-loaded h-BN nanocomposite with PVDF or HL-BN-PVDF (0.23 Scm-1 and 125 mμ), M2: low-loaded h-BN nanocomposite with PVDF or LL-BN-PVDF (0.19 Scm-1 and 80 mμ) and M3: high-loaded h-BN nanocomposite with PVA or HL-BN-PVA (0.36 Scm-1 and 45 mμ) have been used. Three MEAs were fabricated using three membranes of HL-BN-PVDF, LL-BN-PVDF and HL-BN-PVA placed between two APS electrodes denoted as The MEA1, MEA2 and MEA3, respectively, and tested in single AEM electrolyzer cells. The tests are carried out in 1M KOH at 60 °C. As expected, the highest current, 2.6 Acm-2, at 2 V is achieved with MEA3, while MEA2 and MEA1 reached 2.95 and 3 A cm-2, respectively. The performance of AEM water electrolysis with the MEA3 is found to be slightly better than the MEA with the same electrode package but commercial membrane from Fumasep (2.71 Acm-2 at 2.0 V). The stability of the cell with MEA3 is tested under the constant load of 0.5 A cm-2 for 250 h showing negligible degradation. This can be attributed to the high mechanical and chemical stability of the membrane and low dissolution of anodic electrode. Furthermore, the lack of binders or ionomers in the APS electrodes avoids irreversible oxidation reactions of the polymer that can lead to voltage loss over time [3]. Thus, APS electrodes combined with stable nanocomposed (h-BN)-based membranes can potentially lower the cost of the hydrogen produced by water electrolysis. 1. Razmjooei, R. Reißner, A. S. Gago, and A. Ansar, ECS Transactions, 92 (8) 689-702 (2019).2. Wang, T. Weissbach, R. Reißner, A. Ansar, A. S. Gago, S. Holdcroft, K. A. Friedrich, ACS Appl. Energy Mater. DOI: 10.1021/acsaem.9b01392 (2019).3. Li, I. Matanovic, A. S. Lee, E. J. Park, C. Fujimoto, H. T. Chung, and Y. S. Kim, ACS Appl. Mater. Interfaces, 11 (10) 9696-9701 (2019).

Exposure of synthesized Co3O4 nanoparticles to Chlorella minutissima: An ecotoxic evaluation in freshwater microalgae

The current study focuses on the ecotoxicity of cobalt oxide nanoparticles (Co3O4 NPs) in the aquatic environment towards freshwater microalgae, Chlorella minutissima. The interaction of Co3O4 NPs with microalgae shows the growth suppressing effect. The 72 h EC 50 (effective concentration of a chemical having 50% of its impact) values of Co3O4 NPs for C. minutissima was 38.16 ± 1.99 mg/L. The decline in chlorophyll a content and increase in reactive oxygen species (ROS) also indicated the compromised physiological state of microalgae. An increased LDH (lactate dehydrogenase) level in treated samples suggests membrane disintegration by Co3O4 NPs. Light microscopy, scanning electron microscopy (SEM) and Energy Dispersive X-Ray-Scanning electron microscopy (EDX-SEM) further confirm cell entrapment and deposition of Co3O4 NPs on the cell surface. Cellular internalization of NPs, as shown by Inductively Coupled Plasma Atomic Emission Spectroscopy (ICP–AES), also contributes towards the toxicity of NPs. The findings suggest the role of extracellular as well as intracellular nanoparticles (NPs) in exerting a toxic effect on the C. minutissima.

Single-layered GO/LDH hybrid nanoporous membranes with improved stability for salt and organic molecules rejection

Graphene oxide (GO) has been intensively adopted as the building block of separation membranes due to its excellent mechanical strength and atomic scale interlayer spacing. However, the practical application of GO nanosheet-based membranes are limited by the swelling and disintegration of GO membranes in water owing to hydration and electrostatic repulsion. In this work, we firstly fabricated a type of layered double hydroxide (LDH) nanosheets, Co(OH)2 nanosheets, by in-situ growth on the surface of GO nanosheets, followed by coating such GO/Co(OH)2 hybrid nanosheets on a microporous substrate through vacuum suction to form a single-layered GO/Co(OH)2 hybrid thin membrane. The electrostatic and coordination interactions between GO and Co(OH)2 nanosheets are found to stabilize the GO/Co(OH)2 hybrid membrane in water, and the hybrid membrane shows good resistance to shear force in a pressurized crossflow nanofiltration system with crossflow speed as high as 0.5 m s−1. With permeation flux up to 17.0 L m−2 h−1, >91% rejection to inorganic salts like Na2SO4 and >98% rejection to various organic dyes (transmembrane pressure: 6 bar, crossflow speed: 0.27 m s−1), the hybrid membrane also survives the challenges of different pH environments, elevated transmembrane pressure and a 60 h continuous crossflow test. Such significantly improved stability from pure GO membranes is expected to enable the 2D nanomaterial based membranes for practical filtration applications in the water industry.

Antibiofilm activity and cytotoxicity of silk sericin against Streptococcus mutans bacteria in biofilm: an in vitro study.

To investigate the potential of sericin extracted by different methods to inhibit biofilm formation (prevention) and disrupt already formed biofilm (treatment). In this in vitro study, sericin was extracted by heat, acid, alkali and urea. Streptococcus mutans bacteria were cultivated in the presence of various concentrations of sericin to evaluate antibiofilm formation using cell density assay (inhibition effect before biofilm formed). Conversely, various concentrations of sericin were added to a biofilm already formed by Streptococcus mutans bacteria, and the viability of bacteria assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (disruption effects after biofilm formed). Structures of extracted sericin were evaluated using circular dichroism and Fourier-transform infrared spectrometer. The urea-extracted sericin at all concentrations (12.5mg/ml, 25mg/ml, 50mg/ml and 100mg/ml) showed the highest potential antibiofilm activity in terms of both inhibition and disruption effects, compared with sericin extracted by heat, acid or alkali. The heat-extracted and acid-extracted sericin were found to reduce the biofilm formation dose-dependently, while the alkali-extracted sericin did not show either inhibition or disruption effect on the bacterial biofilm. The urea-extracted sericin also killed the bacteria residing within the biofilm, possibly due to its modified structure which may destabilise the bacterial cell wall, leading to membrane disintegration and, finally, cell death. Our results demostrated the antibiofilm activity of sericin. This could form the basis of further research on the mechanism and application of sericin as a novel antibiofilm agent.

Bioactive Secondary Metabolites from Trichoderma spp. against Phytopathogenic Bacteria and Root-Knot Nematode.

Losses in crops caused by plant pathogenic bacteria and parasitic nematode are increasing because of a decrease in efficacy of traditional management measures. There is an urgent need to develop nonchemical and ecofriendly based management to control plant diseases. A potential approach of controlling plant disease in the crops is the use of biocontrol agents and their secondary metabolites (SMs). Luckily fungi and especially the genus Trichoderma comprise a great number of fungal strains that are the potential producer of bioactive secondary metabolites. In this study secondary metabolites from ten Trichoderma spp. were evaluated for their antibacterial and nematicidal potential against phytopathogenic bacteria Ralstonia solanacearum, Xanthomonas compestris and plant parasitic nematode Meloidogyne incognita. Five different growth media were evaluated for the production of SMs. It was shown that SMs of different Trichoderma spp. obtained on different growth media were different in the degree of their bioactivity. Comparison of five growth media showed that SMs produced on solid wheat and STP media gave higher antibacterial activity. SMs of T. pseudoharzianum (T113) obtained on solid wheat media were more effective against the studied bacteria followed by SMs from T. asperelloides (T136), T. pseudoharzianum (T129) and T. pseudoharzianum (T160). Scanning electron microscopy (SEM) was further conducted to observe the effect of SMs on bacterial cell morphology. As evident from the SEM, SMs produced severe morphological changes, such as rupturing of the bacterial cell walls, disintegration of cell membrane and cell content leaking out. SMs from T. viridae obtained on liquid STP and solid wheat media showed the highest percent of M. incognita juveniles (J2s) mortality and inhibition in egg hatching of M. incognita. The results of our study suggest that T. pseudoharzianum (T113) and T. viridae could be selected as an effective candidate for SMs source against phytopathogenic bacteria and M. incognita respectively.

Microwave-Assisted Green Synthesis and Characterization of Silver Nanoparticles Using Melia azedarach for the Management of Fusarium Wilt in Tomato.

These days, research in agriculture is focusing on the theme of sustainability along with protection of agriculture produce. Nanotechnology in the agriculture sector aims for the enhancement of agricultural produce and the reduction of pesticides through providing innovative agrochemical agents and their novel delivery mechanisms. The current investigation involved the green synthesis of silver nanoparticles (AgNPs) from the aqueous leaf extract of Melia azedarach by following a microwave-assisted method to control Fusarium oxysporum, the causal agent of tomato wilt. Biosynthesized Melia leaf extract (MLE)-AgNPs were characterized by UV-visible spectroscopy, Fourier-transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), energy dispersive X-ray (EDX) spectrometry, dynamic light scattering (DLS), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and zeta potential analysis. The intensity of the peak at 434 nm in UV-vis spectra, attributed to the surface plasmon resonance of MLE-AgNPs, changes with reaction parameters. TEM exhibits spherical shaped nanoparticles with an average particle size range from 12 to 46 nm. Efficient inhibition of F. oxysporum, the causal agent of tomato wilt, was achieved after exposure to MLE-AgNPs both in vivo and in vitro. In vitro studies exhibited repressed fungal mycelial growth with 79–98% inhibition as compared to the control. Significant increases in growth parameters of tomato seedlings were observed after treatment with biosynthesized nanoparticles as compared to F. oxysporum-infected plants grown without them under greenhouse conditions. Furthermore, SEM imaging was done to reveal the prominent damage on the cell wall of hyphae and spores after MLE-AgNP treatment. Propidium iodide (PI) staining of mycelium indicated the extent of cell death, causing irretrievable damage and disintegration of cellular membranes by altering the membrane permeability. Also, 2′,7′-dichlorofluorescin diacetate (DCFH-DA) fluorescence specifies intracellular reactive oxygen species (ROS) production in F. oxysporum after treatment with MLE-AgNPs. The current investigation suggested that biosynthesized nanoparticles can revolutionize the field of plant pathology by introducing an environment-friendly approach for disease management and playing a potential part in agriculture industry. However, to date, little work has been done to integrate nanotechnology into phytopathology so, this area of research is in need of adoption and exploration for the management of plant diseases.