Loss Of Cell Integrity Research Articles

Characterization of the Aluminium-Based Metal Foam Properties for Automotive Applications

AbstractMetal foams are solids where the gas is filled inside uniformly in the metal matrix. Blowing agent supplies air inside the parent metal, and metal foam has emerged to be a promising material because of its low density, high absorption capacity, low thermal conductivity and high strength which finds its huge applications in automobile components. The present work deals with the application of the aluminium metal foam with different densities 200 and 400 kg/m3 in automobiles. Various tests such as toughness, hardness, bending and compression are carried out for four chosen densities, and the values are compared with the aluminium base metal. The result showed that the hardness value increased significantly by 24.48% with the rise in the density from 200 to 400 kg/m3. Maximum modulus of resilience for the low-density specimen is found to be 2.21 MJ/m3. Surface topography showed irregular pore shapes with discontinuity, resulting in a loss of cell integrity with the neighbouring cell walls. This affected the performance of the foam significantly. Thermal experiments were carried out to determine the thermal conductivity where thermal conductivity increased by 122% with the rise in the density from 200 to 400 kg/m3. Based on the results, it is concluded that aluminium foam with density 400 kg/m3 can be recommended for use in automobile applications due to its lightweight properties, which contribute to improving fuel efficiency, impact absorption capacity and the vehicle’s speed. Additionally, the air trapped within the foam cells serves as a sound barrier and insulator in cars.

THE ANTIFUNGAL EFFECTIVITY TEST OF ETHANOLIC EXTRACT OF KIDNEY BEAN (PHASEOLUS VULGARIS L.) AGAINST CANDIDA ALBICANS USING IN VITRO METHOD

Introductions: Candida albicans can causes an infection called candidiasis. Kidney beans (Phaseolus vulgaris L.) are natural products that contain several active compounds such as saponins, flavonoids, tannins, and coumarins that can damage the membrane and cell wall of Candida albicans. Methods: The design of this study used true experimental post control design only with the tube dilution method. Extract concentrations used were 3200 mg/ml, 3600 mg/ml, 4000 mg/ml, 4400 mg/ml, 4800 mg/ml, 5200 mg/ml, 5600 mg/ml, and 6000 mg/ml. This study used a negative control (extract 0%) and a positive control (Nystatin 1%), each of which was added with Candida albicans. Results: The results showed that the Minimum Inhibitory Concentration (MIC) was obtained at 5200 mg/ml and the Minimum Bactericidal Concentration (MBC) was obtained at 6000 mg/ml. One Way ANOVA statistical test results showed a significant difference (significance value 0,000; p< 0.05). Pearson correlation test showed the direction of the negative correlation (r = -0.945; p <0.01). Discussion: The ability of ethanolic extract of kidney bean to inhibit growth and kill the Candida albicans is due to the active compounds contained in it, namely flavonoids, saponins, tannins, and coumarin. Flavonoids can cause protein denaturation. Saponins have the ability to kill Candida albicans by forming pores in the double lipidic layer that results on the loss of cell integrity. Tannins have the ability to inhibit fungal growth by inhibiting the synthesis of chitin and damaging cell membranes. Coumarins can cause fungal death by interfering with cell wall synthesis. Conclusions: The conclusion of this research is the ethanol extract of kidney bean can inhibit at the concentration of 5200 mg/ml and kill the growth of Candida albicans at the concentration of 6000 mg/ml.

The competitive advantage of Microcystis aeruginosa over Scenedesmus obliquus weakened by exposure to polylactic acid microplastics

Aquatic ecosystems are heavily affected by microplastics (MPs), and its impacts on aquatic life have received extensive attention. However, it is still unclear how biodegradable MPs influence the growth and competition of phytoplankton. In this study, the response of growth dynamics, alternation in algal cell morphology and toxin-producing capability, and changes in the extracellular process of Microcystis aeruginosa (M. aeruginosa) and Scenedesmus obliquus (S. obliquus) were systematically studied in monoculture and co-culture conditions with and without the presence of polylactic acid MPs (PLA-MPs). The results indicated that although the loss of cell integrity was observed, PLA-MPs addition (50 mg/L) caused a 1.40- and 1.36-fold increase in cell densities of M. aeruginosa and S. obliquus in monoculture systems, respectively. This suggests the PLA-MPs more favored the growth of M. aeruginosa. This effect was manifested in co-culture conditions, because the degradation of PLA-MPs provided additional inorganic carbon in the experimental systems and benefited the growth of both M. aeruginosa and S. obliquus. Meanwhile, the synthesis of microcystins from the toxic M. aeruginosa was substantially reduced upon PLA-MPs exposure, which reduced the competitive advantage of M. aeruginosa over S. obliquus. Thus; the cell density of M. aeruginosa and S. obliquus showed 1.15- and 1.88-folds increasing compared to that without PLA-MPs addition. This interaction between PLA-MPs and algae weakened the competitive advantage of M. aeruginosa over S. obliquus, but their competitive outcomes remained unchanged. The results provided new insights into understanding the potential ecological risks of biodegradable plastics in aquatic ecosystems.

Unveiling the efficacy of pre-emergent application of young Eucalyptus globulus leaves as a weed control strategy: Bridging macroscopic effects and cellular responses

Allelopathy, the inhibition of neighbouring plant growth by certain plants, can be particularly useful if applied in a targeted way for weed management. So, this study aimed to assess and characterize the herbicidal activity of fresh and dried leaves from young Eucalyptus globulus Labill. trees applied as a soil amendment. For this, fresh and dried leaves (FL and DL, respectively) were incorporated into the soil at different concentrations (0, 1, 5, and 10% w/w), where Portulaca oleracea L. seeds were sown. After 5 weeks of exposure, results revealed that the soil incorporation of DL at 10% (w/w) presented the strongest herbicidal properties, inhibiting seed germination by 63% and inducing the loss of cell viability. To unravel the possible mode of action and the main targets at both cellular and subcellular levels, an in vitro experiment was performed. Purslane seeds were sown in a nutritive medium containing different dilutions of an aqueous extract prepared with dried eucalyptus leaves. After 5 days of exposure, germinated seedlings were processed for transmission electron microscopy and histological analyses as well as for reactive oxygen species (ROS) in vivo detection by confocal laser scanning microscopy. Results revealed that the allelochemical release from DL induced ROS overproduction, resulting in the loss of cell integrity and organization, which was characterized by damage to several cellular sub-structures, along with enhanced accumulation of lipid droplets. Overall, the incorporation of DL into the soil can represent a sustainable alternative to reduce synthetic herbicide application and subsequent environmental contamination.

Effect of pre-chlorination on Microcystis aeruginosa cell integrity and microcystins removal

ABSTRACT High nutrient input into waterbodies increases potentially toxic cyanobacteria blooms. Pre-chlorination treatment is excellent for cyanobacterial cell inactivation and dissolved cyanotoxins removal in water supply sources. This study aims to evaluate the pre-chlorination effect treatment on Microcystis aeruginosa cell integrity and its hepatotoxins (microcystins, MC) release and degradation on a laboratory scale. Loss of cell integrity is observed with increasing chlorine concentration. The maximum cell permeability without cell disintegration occurs at 2.5 mg Cl2 L−1. Degradations up to 70% and chlorophyll-a reductions over 70% are observed after 60 min exposure, with complete degradation at 2.5 and 3 mg Cl2 L−1. However, chlorine (Cl2) exposure over 40.66 mg min L−1 is required for microcystins MC-LR (i.e. radicals: leucine (L) and arginine (R)) and MC-RR (i.e. radicals: arginine and arginine) oxidation oxidation to concentrations below the legislation value for water potability.

BI-Y, an Neuropilin-1 Antagonist, Enhances Revascularization and Prevents Vascular Endothelial Growth Factor-A Induced Retinal Hyperpermeability in Rodent Models of Retinopathies.

Diabetic retinopathy (DR) is a leading cause of vision loss in working-age adults. Despite an established standard of care for advanced forms of DR, some patients continue to lose vision after treatment. This may be due to the development of diabetic macular ischemia (DMI), which has no approved treatment. Neuropilin-1 (Nrp-1) is a coreceptor with two ligand-binding domains, with semaphorin-3A (Sema3A) binding to the A-domain and vascular endothelial growth factor-A (VEGF-A) binding to the B-domain. Sema3A directs a subset of neuronal growth cones as well as blood vessel growth by repulsion; when bound to Nrp-1, VEGF-A mediates vascular permeability and angiogenesis. Modulating Nrp-1 could therefore address multiple complications arising from DR, such as diabetic macular edema (DME) and DMI. BI-Y is a monoclonal antibody that binds to the Nrp-1 A-domain, antagonizing the effects of the ligand Sema3A and inhibiting VEGF-A-induced vascular permeability. This series of in vitro and in vivo studies examined the binding kinetics of BI-Y to Nrp-1 with and without VEGF-A165, the effect of BI-Y on Sema3A-induced cytoskeletal collapse, the effect of BI-Y on VEGF- A165-induced angiogenesis, neovascularization, cell integrity loss and permeability, and retinal revascularization. The data show that BI-Y binds to Nrp-1 and inhibits Sema3A-induced cytoskeletal collapse in vitro, may enhance revascularization of ischemic areas in an oxygen-induced retinopathy mouse model, and prevents VEGF-A-induced retinal hyperpermeability in rats. However, BI-Y does not interfere with VEGF-A-dependent choroidal neovascularization. These results support further investigation of BI-Y as a potential treatment for DMI and DME. SIGNIFICANCE STATEMENT: Diabetic macular ischemia (DMI) is a complication of diabetic retinopathy (DR) with no approved pharmacological treatment. Diabetic macular edema (DME) commonly co-occurs with DMI in patients with DR. This series of preclinical studies in mouse and rat models shows that neuropilin-1 antagonist BI-Y may enhance the revascularization of ischemic areas and prevents vascular endothelial growth factor-A (VEGF-A)-induced retinal hyperpermeability without affecting VEGF-A-dependent choroidal neovascularization; thus, BI-Y may be of interest as a potential treatment for patients with DR.

Application of Digital Holographic Microscopy to Analyze Changes in T-Cell Morphology in Response to Bacterial Challenge

Quantitative phase imaging (QPI) is a non-invasive, label-free technique used to detect aberrant cell morphologies caused by disease, thus providing a useful diagnostic approach. Here, we evaluated the potential of QPI to differentiate specific morphological changes in human primary T-cells exposed to various bacterial species and strains. Cells were challenged with sterile bacterial determinants, i.e., membrane vesicles or culture supernatants, derived from different Gram-positive and Gram-negative bacteria. Timelapse QPI by digital holographic microscopy (DHM) was applied to capture changes in T-cell morphology over time. After numerical reconstruction and image segmentation, we calculated single cell area, circularity and mean phase contrast. Upon bacterial challenge, T-cells underwent rapid morphological changes such as cell shrinkage, alterations of mean phase contrast and loss of cell integrity. Time course and intensity of this response varied between both different species and strains. The strongest effect was observed for treatment with S. aureus-derived culture supernatants that led to complete lysis of the cells. Furthermore, cell shrinkage and loss of circular shape was stronger in Gram-negative than in Gram-positive bacteria. Additionally, T-cell response to bacterial virulence factors was concentration-dependent, as decreases in cellular area and circularity were enhanced with increasing concentrations of bacterial determinants. Our findings clearly indicate that T-cell response to bacterial stress depends on the causative pathogen, and specific morphological alterations can be detected using DHM.

Inhibitory effect and mechanism of curcumin-based photodynamic inactivation on patulin secretion by Penicillium expansum

Patulin (PAT) contamination caused by Penicillium expansum growth in postharvest fruit brings safety problems. This study investigated the inhibitory effect and mechanism of curcumin (CUR)-based photodynamic inactivation (PDI) on PAT secreted by P. expansum in vitro using a multifunctional light source instrument. After being treated with photosensitive CUR, the growth of mycelium was completely inhibited on potato dextrose agar medium, and the maximum PAT reduction of 93.06% can be achieved (330.0 J/cm2, 300 μM CUR) in liquid medium. The inherent mechanism was that PDI causes the reactive oxygen species accumulation in cells, which destroyed the antioxidant defense system and induced severe oxidative damage, resulting in the loss of cell integrity. Furthermore, mycelial apoptosis caused up-regulation of five genes, including PatB, PatE, PatG, PatI, and PatL, but down-regulated ten genes in the PAT biosynthetic pathway. These findings suggest that CUR-based PDI can inhibit fungal infection and toxin secretion. Industrial relevanceThis study found that photosensitized curcumin can significantly inhibit mycelial growth and the secretion of patulin by Penicillium expansum. Curcumin-based photodynamic inactivation has the feasibility of the application on fresh fruits and vegetables, such as apples or oranges, and can avoided excessive waste of resources in picking, transporting, storing, and selling.

A multistep in vitro hemocompatibility testing protocol recapitulating the foreign body reaction to nanocarriers

The development of drug nanocarriers based on polymeric, lipid and ceramic biomaterials has been paving the way to precision medicine, where the delivery of poorly soluble active compounds and personalized doses are made possible. However, the nano-size character of these carriers has been demonstrated to have the potential to elicit pathways of the host response different from those of the same biomaterials when engineered as larger size implants and of the drugs when administered without a carrier. Therefore, a specific regulatory framework needs to be made available that can offer robust scientific insights and provide safety data by reliable tests of these novel nano-devices. In this context, the present work presents a multistep protocol for the in vitro assessment of the hemocompatibility of nanocarriers of different physicochemical properties. Poly (ethyl butyl cyanoacrylate) nanoparticles and lipid-based (LipImage™ 815) nanoparticles of comparable hydrodynamic diameter were tested through a battery of assays using human peripheral blood samples and recapitulating the main pathways of the host response upon systemic administration; i.e., protein interactions, fibrinogen-platelet binding, cytotoxicity, and inflammatory response. The data showed the sensitivity and reproducibility of the methods adopted that were also demonstrated to determine individual variability as well as to discriminate between activation of pathways of inflammation and unintended release of inflammatory signaling caused by loss of cell integrity. Therefore, this multistep testing is proposed as a reliable protocol for nanoparticle development and emerging regulatory frameworks.Graphical abstract

Enhanced antitumor effect of L-buthionine sulfoximine or ionizing radiation by copper complexes with 2,2´-biquinoline and sulfonamides on A549 2D and 3D lung cancer cell models.

Two ternary copper(II) complexes with 2,2'-biquinoline (BQ) and with sulfonamides: sulfamethazine (SMT) or sulfaquinoxaline (SDQ) whose formulae are Cu(SMT)(BQ)Cl and Cu(SDQ)(BQ)Cl·CH3OH, in what follows SMTCu and SDQCu, respectively, induced oxidative stress by increasing ROS level from 1.0μM and the reduction potential of the couple GSSG/GSH2. The co-treatment with L-buthionine sulfoximine (BSO), which inhibits the production of GSH, enhanced the effect of copper complexes on tumor cell viability and on oxidative damage. Both complexes generated DNA strand breaks given by-at least partially-the oxidation of pyrimidine bases, which caused the arrest of the cell cycle in the G2/M phase. These phenomena triggered processes of apoptosis proven by activation of caspase 3 and externalization of phosphatidylserine and loss of cell integrity from 1.0μM. The combination with BSO induced a marked increase in the apoptotic population. On the other hand, an improved cell proliferation effect was observed when combining SDQCu with a radiation dose of 2Gy from 1.0μM or with 6Gy from 1.5μM. Finally, studies in multicellular spheroids demonstrated that even though copper(II) complexes did not inhibit cell invasion in collagen gels up to 48h of treatment at the higher concentrations, multicellular resistance outperformed several drugs currently used in cancer treatment. Overall, our results reveal an antitumor effect of both complexes in monolayer and multicellular spheroids and an improvement with the addition of BSO. However, only SDQCu was the best adjuvant of ionizing radiation treatment.

Traganum nudatum Possesses Anticancer Efficacy against Breast Cancer MDA-MB-231 Cells

Background: Medicinal plants are effective sources for both traditional and modern medicines; herbal medicine has been shown to have genuine utility. 80% of rural population depends on natural products as primary health care. Traganum nudatum (T. nudatum) is a medicinal plant widely used in alternative medicine. T. nudatum has not been greatly investigated for its anticancer potential.
 Aim: This study aimed to evaluate the in vitro anticancer efficacy of T. nudatum against breast cancer cell line; MDA-MB-231.
 Materials and Methods: Extracts of T. nudatum were evaluated to assess their in vitro cytotoxicity against the MDA-MB-231 breast cancer cell line. Leaves, flowers, and stems of T. nudatum were extracted by using 70 % ethanol. the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays were used to evaluate the in vitro cytotoxicity of these extracts against MDA-MB-231 cell line. The apoptotic cellular morphological changes were investigated using inverted and fluorescence microscopes.
 Results: The results showed that T. nudatum extracts exhibited anticancer activity with a half-maximal inhibitory concentration (IC50) of 150 µg/mL against MDA-MB-231 cell line. Loss of cell integrity, shrinkage of the cytoplasm, and cell detachment were observed in the extract-treated MDA-MB-231 cells.
 Conclusion: T. nudatum extracts showed cytotoxic effects against MDA-MB-231 cell line. It may be used as a future medication to cure breast cancer.

Antifungal Activity and Mechanism of Action of Different Parts of Myrtus communis Growing in Saudi Arabia against Candida Spp.

Discovering new antifungal drugs from natural products is a key target for the treatment of infections, such as candidiasis and other Candida-related infections. As current therapeutic drugs for the treatment of infections, such as candidiasis and other Candida-related infections, have adverse effects on human health, discovering new antifungal drugs from natural products is urgently needed. The objective of this study was to evaluate the antifungal activity of the methanolic and sodium phosphate buffer extracts derived from various parts of Myrtus communis, a plant that is traditionally used in Saudi Arabia, against Candida albicans (ATCC 10213), Candida glabrata (ATCC 2001), Candida kefyr (ATCC 66028), Candida parapsilosis (ATCC 22019), and Candida tropicalis (ATCC 750). A well diffusion assay was performed to assess the antifungal activity through the measurement of the zone of inhibition. Of the extracts, those extracted with methanol from the roots and leaves displayed strong inhibitory activity against Candida glabrata ( 23.5 ± 0.12 and 20.7 ± 0.22 , respectively), at 50 mg/ml, with 5 mg/ml fluconazole administered as the standard control. The minimal inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values were 12.5 mg/ml and 25 mg/ml for the M. communis root extract and 25 mg/ml and 50 mg/ml for the M. communis leaf extract against Candida glabrata. The results were confirmed by scanning electron microscopy (SEM) imaging of the control and treated strains of Candida glabrata. Based on SEM, these extracts could alter the morphology and cause loss of cell integrity. The effect of M. communis root and leaf extracts on Candida cells was also determined by measuring the absorbance at 260 nm after treatment for 1 h at 37°C. Interestingly, the 260 nm absorbing material was higher in Candida glabrata than in the resistant strain, Candida parapsilosis (ATCC 22019). Based on our findings, the crude methanolic extract of M. communis roots and leaves exhibited good antifungal activity against the Candida glabrata strain. SEM results and estimation of the 260 nm absorbance material proved that the extract might act on the cell wall and cell membrane of Candida cells, further leading to cell death.

Antibacterial activity of ZnO quantum dots and its protective effects of chicks infected with Salmonella pullorum

In light of emerging antibiotic resistance, synthesis of active, environmental friendly antimicrobial alternatives becomes increasingly necessary. In this study, ZnO quantum dots (ZnO QDs) were developed by the sol–gel method and characterized. The antibacterial activities of ZnO QDs against Escherichia coli (E. coli), Staphylococcus aureus (S. aureus) and Salmonella Pullorum (S. Pullorum) were systematically investigated. Moreover, the protective effects of ZnO QDs on Salmonella-caused pullorosis in chicks were also explored. The results indicated that the size range of ZnO QDs was 3–6 nm. Antibacterial results showed that ZnO QDs treatment inhibited the growth of E. coli, S. aureus, and S. Pullorum in the rate of 87.06 ± 0.98%, 94.75 ± 2.28%, and 85.55 ± 1.15%, respectively. Its excellent antibacterial property was manifested with the minimum inhibitory concentration of 0.7812, 0.0976, and 0.1953 mg ml−1, which may be attributed to the production of reactive oxygen species, the dissolution of Zn2+ ions, and the loss of cell integrity. Furthermore, in the in vivo test, the ZnO QDs effectively reduced the mortality of chicks infected with S. Pullorum via regulating the balance of the intestinal flora, protecting liver and intestine, and modulating the balance of antioxidation systems. This study reveals that ZnO QDs exerts remarkably antibacterial activity in vitro and anti-pullorosis effect in chicks.

Protective Effect of Emblica officinalis in Cyprinus carpio against Hepatotoxicity Induced by Malachite Green: Ultrastructural and Molecular Analysis

Malachite green (MG) dye, besides coloring is used as an effective aquaculture therapeutic. The present study assesses the mitigating ability of Emblica officinalis (EO) fruit extract against the dye induced chronic (60 days) cyto-toxicity in Cyprinus carpio. For this, four experimental groups were maintained: group I—control, group II—MG, group III—EO (positive control), group IV—MG + EO. The study was made at three tiers: detailing structural anomalies using a light microscope and transmission electron microscope (TEM), biochemical estimation of antioxidant enzymes, and lipid peroxidation and molecular analysis of expression patterns of HSP70, and CYP1A genes. MG intoxication resulted in necrosis, cytoplasmic vacuolation, glycogen depletion, abundant macrophages, loss of cell integrity and prominent nuclear alterations. Significant (p < 0.05) inhibition in the activities of catalase (CAT), superoxide dismutase (SOD), glutathione-S-transferase (GST) and reduced glutathione (GSH), along with an elevation in malondialdehyde (MDA) levels, occurred after 60 days of MG exposure. CYP1A and HSP70 genes presented a significant change in their expression in MG treated fish. Whereas oral supplementation with EO significantly restored the histo-architecture, normalized the altered enzymatic activity, reduced the oxidative stress level and regulated the expression of HSP70 and CYP1A genes. Thus, it can be concluded that EO acted as an effective ameliorant against malachite green induced cyto-toxicity in Cyprinus carpio.

Use of optical coherence tomography and light microscopy for characterisation of mechanical properties and cellular level responses of ‘Centurion’ blueberries during weight loss

Postharvest blueberry softening hinders consumer acceptance and correlates with high moisture loss during storage. Such textural variations have been attributed to factors such as turgor, cell wall modifications and other microstructural changes in the outer cell layers of the fruit. This paper investigates the impact of moisture loss on blueberry quality, as well as the structure and/or function relationships associated with fruit mechanical properties during postharvest using an integrated physical and novel microstructural approach. Four different weight loss conditions [62%, 76%, 93% and 98% relative humidity (RH)] at 5.7 °C were evaluated over a three-week postharvest storage period to assess blueberry mechanical parameters using texture profile analysis (TPA), whilst simultaneously assessing microstructural changes non-destructively by optical coherence tomography (OCT) and destructively by light microscopy. Water loss affected postharvest blueberry cell morphology and resulting texture profile. Increasing weight loss resulted in a reduction in hardness, hardness slope and chewiness but an increase in cohesiveness and springiness. Blueberries stored at 98% RH also showed increasing hardness after storage, as opposed to the softening of fruit observed for the other RH conditions. The 3D OCT images revealed that the layer thickness of the top two cellular layers in the skin of blueberries increased with increasing weight loss as a result of moisture loss and had linear Pearson's correlations with TPA parameters of blueberry. Microscopy more clearly illustrated irregular cell boundaries, increased intercellular spaces, loss of cell integrity and cell collapse during moisture loss. Future work should investigate the chemical compositional changes in the epidermal and sub-epidermal layers of the blueberry stored at low water loss conditions in order to investigate the observed firming effect over the storage period. OCT has the potential to enable non-destructive real-time monitoring of the near-surface internal cellular layers of blueberries. Future work is required to validate the use of OCT as a non-destructive tool for this purpose.

Combined impact of silver nanoparticles and chlorine on the cell integrity and toxin release of Microcystis aeruginosa

Silver nanoparticles (AgNPs) have shown to be toxic to freshwater cyanobacterial species, and sodium hypochlorite (NaOCl) is a common oxidant for the treatment of cyanobacterial cells. AgNPs have a high possibility of co-existing with the cyanobacterial cells in the aqueous environments leading to its exposure to NaOCl during water treatment; however, their combined effects on the cyanobacterial cells are largely undocumented. This work compares the individual and combined effect of AgNP and NaOCl on the integrity and toxin (microcystins) release of Microcystis aeruginosa at varying levels. The results show that the AgNP (0.2–0.6 mg/L) alone has negligible effects on the cell lysis, while NaOCl alone shows concentration-dependent (0.2 < 0.4 < 0.6 mg/L) rupturing of cells. In contrast, the AgNP + NaOCl (0.2–0.6 mg/L) samples show increasing loss in cell integrity at higher AgNP (0.4 and 0.6 mg/L) levels than the NaOCl only samples. NaOCl exposure results in increasing dissolution of AgNPs with time, releasing silver ions (Ag+), affecting its size and morphology. The cell-associated total Ag declines over time with an increase in NaOCl levels, maybe due to increasing cell-lysis or NaOCl induced oxidative dissolution of AgNPs. The cell-associated total Ag and released Ag+ possibly weaken the cellular membrane, thus assisting NaOCl in faster cell-lysis. The combined exposure of AgNP and NaOCl also results in a higher release of toxin from the cells. This work collectively reveals that the AgNPs combined with NaOCl can enhance the cell lysis and release of toxins.

SWI/SNF inactivation in the endometrial epithelium leads to loss of epithelial integrity.

Although ARID1A mutations are a hallmark feature, mutations in other SWI/SNF (SWItch/Sucrose Non-Fermentable) chromatin remodeling subunits are also observed in endometrial neoplasms. Here, we interrogated the roles of Brahma/SWI2-related gene 1 (BRG1, SMARCA4), the SWI/SNF catalytic subunit, in the endometrial epithelium. BRG1 loss affects more than one-third of all active genes and highly overlaps with the ARID1A gene regulatory network. Chromatin immunoprecipitation studies revealed widespread subunit-specific differences in transcriptional regulation, as BRG1 promoter interactions are associated with gene activation, while ARID1A binding is associated with gene repression. However, we identified a physiologically relevant subset of BRG1 and ARID1A co-regulated epithelial identity genes. Mice were genetically engineered to inactivate BRG1 specifically in the endometrial epithelium. Endometrial glands were observed embedded in uterine myometrium, indicating adenomyosis-like phenotypes. Molecular similarities were observed between BRG1 and ARID1A mutant endometrial cells in vivo, including loss of epithelial cell adhesion and junction genes. Collectively, these studies illustrate overlapping contributions of multiple SWI/SNF subunit mutations in the translocation of endometrium to distal sites, with loss of cell integrity being a common feature in SWI/SNF mutant endometrial epithelia.

Mycoplasma bovis-generated reactive oxygen species and induced apoptosis in bovine mammary epithelial cell cultures

Mycoplasma bovis is an important cause of bovinemastitis in China and worldwide. We hypothesized that M. bovis damages bovine mammary epithelial cells (bMEC), with the degree of damage varying among field isolates. Our objective was to evaluate 2 novel sequence type (ST) field strains of M.bovis (ST172 and ST173) for their ability to induce oxidative stress, cytotoxicity, pathomorphological changes, and apoptosis in bMEC, as a model for pathogenesis of M. bovis-induced bovine mastitis. Cytotoxicity (as indicated by release of lactate dehydrogenase, LDH) from bMEC depended on multiplicity of infection (MOI), with a high MOI (1:1,000) being required to induce cytotoxicity. Morphological changes in bMEC, including shrinkage, loss of cell integrity, and heavy staining (hematoxylin and eosin) of cytoplasm were apparent 24 h after infection with ST172 or ST173 M.bovis, with more severe changes being induced by the latter strain. Adhesion and invasion assays both had curvilinear patterns, peaking 12 h after infection with MOI of 1:1,000. Both production of reactive oxygen species (ROS) and proportion of apoptotic cells increased with time after infection. Increased Bax/Bcl-2 ratios and activation of caspase-3 implied involvement of mitochondria-dependent pathways ofapoptosis. Furthermore, intracellular ROS generation, apoptosis, and cleaved caspase-3 were mitigated by N-acetyl-l-cysteine, a ROS scavenger. Both interleukin (IL)-1β and IL-6 were significantly upregulated by ST172 and ST173 M. bovis, with little change in expression of tumor necrosis factor-α. One ST173 M. bovis isolate had the greatest cytotoxicity of all of our field isolates, with the highest LDH release, adhesion, invasion, ROS production, and apoptosis. In conclusion, our hypothesis was supported: M. bovis damaged bMEC by generating ROS and initiating a mitochondria-dependent pathway ofapoptosis, with the degree of damage varying among field isolates. This study provided new knowledge regarding pathogenesis of M. bovis-induced bovine mastitis.

Antibacterial properties and mechanism of biopolymer-based films functionalized by CuO/ZnO nanoparticles against Escherichia coli and Staphylococcus aureus

In this study, the nanocomposite film (SA-CS@CuO/ZnO) composed of sodium alginate (SA) and chitosan (CS) functionalized by copper oxide nanoparticles (CuONPs) and zinc oxide nanoparticles (ZnONPs) was fabricated, then its antibacterial mechanisms against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) were systematically investigated. When the contents of CuONPs and ZnONPs reached 1.5 % (w/w) and 0.5 % (w/w), respectively, the SA-CS@CuO/ZnO exhibited great mechanical, barrier, and optical properties. Moreover, the incorporation of ZnONPs enhanced the photocatalytic ability of SA-CS@CuO/ZnO, producing a high level of reactive oxygen species under light irradiation. Further, antibacterial results showed that SA-CS@CuO/ZnO treatment inhibited the growth of E. coli and S. aureus higher than 60 % in the dark and exceeded 90 % under light irradiation. This was also manifested in the incompleteness of bacterial cell structure, accompanied by unstable cellular redox balance and DNA disruption. The functions of differentially expressed genes screened by transcriptome analysis were mainly involved in membrane transport, cell wall and membrane synthesis, cellular antioxidant defense system, cell membrane and DNA repair system. The changes in bacterial transcriptional regulation reflected the disturbance in the physiological activities and loss of cell integrity, leading to damage of bacterial cells or death.

Anticancer Potential of Pulicaria crispa Extract on Human Breast Cancer MDA-MB-231 Cells

Background: Breast cancer is the common cause of deaths among women globally with 15% mortality globally. Introduction: Today, about 80% of the rural population depends on natural products as primary health care. Pulicaria crispa (L., family Compositae) is utilized in traditional medicine for curing colds, coughs, colic, and excessive sweating and as a carminative. Methods: The extracts of Pulicaria crispa; grown in Saudi Arabia; were assessed to measure the cytotoxicity with MDA-MB-231 breast cancer cell lines. Soxhlet extraction was utilized for stem, leaves and flower with 70% ethanol. The cytotoxicity of the extracts with MDA-MB-231 breast cancer cells was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays. Results: The apoptotic cellular morphological alterations were detected by fluorescence microscopes. The results indicated that Pulicaria crispa exhibited a strong anticancer activity with a halfmaximal inhibitory concentration (IC50) of 180 µg/mL against breast cancer cells. The loss in cell integrity, shrinkage of cytoplasm, and cell detachment were seen in the extract treated with MDAMB- 231 cells. The cell death was due to membrane destruction. Conclusion: Pulicaria crispa extracts indicated significant cytotoxicity against human breast cancer cells (MDA-MB-231 cells). The extract of this plant may be given to the patients having breast cancer.