Concentration-dependent Reduction Research Articles

The inhibitory effect of Sunset Yellow on thermally induced Human Serum Albumin aggregates: Possible role in naturopathy

Intensive research in the field of protein aggregation confirmed that the deposition of amyloid fibrils of proteins are the major cause for the development of various neurotoxic and neurodegenerative diseases, which could be controlled by ensuring the efficient inhibition of aggregation using anti aggregation strategies. Herein, we elaborated the anti amyloidogenic potential of Sunset Yellow (SY) dye against Human Serum Albumin (HSA) fibrillogenesis utilising different biophysical, computational and microscopic techniques. The inhibitory effect of sunset yellow was confirmed by Rayleigh Light Scattering (RLS) measurements along with different dye binding assays (ANS, ThT and CR) by showing concentration dependent reduction in scattering intensity and fluorescence intensity respectively. Further, destabilization and anti fibrillation activity of HSA aggregates were characterized through spectroscopic techniques like Circular Dichroism (CD) and other microscopic techniques like Transmission Electron Microscopy (TEM) for elucidating the structural properties. The SDS-PAGE was also carried out that render the disaggregation effect of the dye on the protein. Moreover, Molecular Docking studies revealed the binding parameters justifying the stable protein-dye complex. Simulation studies were also performed accordingly. Thus, this dye which is used as food additive can serve as a potential aggregation inhibiting agent that can aid in the prevention of amyloidogenic diseases.

Protective Effect of Bergapten against Human Erythrocyte Hemolysis and Protein Denaturation In Vitro.

Bergapten, a furocoumarin found in many medicinal plants, is used for the management of various conditions. The present in vitro study evaluated the ability of bergapten to prevent human erythrocyte hemolysis and protein denaturation. Bergapten administered at 10, 30, and 100 μg/ml exhibited a significant concentration-dependent protection on the erythrocyte membrane exposed to hypotonicity and heat-induced hemolysis. The concentration at which bergapten inhibited 50% of the cells from hemolysis (IC50) was determined on a dose-response curve, plotted as logarithmic (concentration) against percentage inhibition, keeping the hemolysis produced within the control group at 100%. Bergapten treatment produced an IC50 value of 7.71 ± 0.27 μg/ml and 4.23 ± 0.42 μg/ml for hypotonicity and heat-induced hemolysis, respectively. Diclofenac sodium at similar concentrations produced an IC50 value of 12.22 ± 0.30 μg/ml and 9.44 ± 0.23 μg/ml in the hypotonicity and heat-induced hemolysis, respectively. The ability of bergapten to inhibit protein denaturation was studied as part of an investigation on its mechanism of action. The results showed a significant concentration-dependent reduction in protein denaturation. When administered at 10, 30, and 100 μg/ml, bergapten produced a concentration-dependent reduction in albumin denaturation. Bergapten inhibited protein denaturation with IC50 values of 5.34 ± 0.30 μg/ml and 12.18 ± 0.20 μg/ml in the heat-treated egg albumin and bovine serum albumin denaturation experiments, respectively. Diclofenac sodium (10, 30, and 100 μg/ml) exhibited a similar protection against heat-treated egg albumin and bovine serum albumin denaturation experiments with IC50 values of 8.93 ± 0.17 μg/ml and 12.72 ± 0.11 μg/ml, respectively. Taken together, data from this study show that the pharmacological properties of bergapten may in part be related to its membrane-stabilizing and antidenaturation properties.

Acute toxicity of methomyl commercial formulation induces morphological and behavioral changes in larval zebrafish (Danio rerio)

The use of pesticides has continue grown over recent years, leading to several environmental and health concerns, such as the contamination of surface and groundwater resources and associated biota, potentially affecting populations that are not primary targets of these complex chemical mixtures. In this work, we investigate lethal and sublethal effects of acute exposure of methomyl commercial formulation in zebrafish embryo and larvae. Methomyl is a broad-spectrum carbamate insecticide and acaricide that acts primarily in acetylcholinesterase inhibition (AChE). Methomyl formulation 96 h-LC50 was determined through the Fish Embryo Acute Toxicity Test (FET) and resulted in 1.2 g/L ± 0.04. Sublethal 6-day exposure was performed in six methomyl formulation concentrations (0.5; 1.0; 2.2; 4.8; 10.6; 23.3 mg/L) to evaluate developmental, physiological, morphological, behavioral, biochemical, and molecular endpoints of zebrafish early-development. Methomyl affected embryo hatching and larva morphology and behavior, especially in higher concentrations; resulting in smaller body and eyes size, failure in swimming bladder inflation, hypolocomotor activity, and concentration-dependent reduction of AChE activity; demonstrating methomyl strong acute toxicity and neurotoxic effect.

Linking In Vitro Models of Endothelial Dysfunction with Cell Senescence.

Endothelial cell dysfunction is the principal cause of several cardiovascular diseases that are increasing in prevalence, healthcare costs, and mortality. Developing a standardized, representative in vitro model of endothelial cell dysfunction is fundamental to a greater understanding of the pathophysiology, and to aiding the development of novel pharmacological therapies. We subjected human umbilical vein endothelial cells (HUVECs) to different periods of nutrient deprivation or increasing doses of H2O2 to represent starvation or elevated oxidative stress, respectively, to investigate changes in cellular function. Both in vitro cellular models of endothelial cell dysfunction-associated senescence developed in this study, starvation and oxidative stress, were validated by markers of cellular senescence (increase in β-galactosidase activity, and changes in senescence gene markers SIRT1 and P21) and endothelial dysfunction as denoted by reductions in angiogenic and migratory capabilities. HUVECs showed a significant H2O2 concentration-dependent reduction in cell viability (p < 0.0001), and a significant increase in oxidative stress (p < 0.0001). Furthermore, HUVECs subjected to 96 h of starvation, or exposed to concentrations of H2O2 of 400 to 1000 μM resulted in impaired angiogenic and migratory potentials. These models will enable improved physiological studies of endothelial cell dysfunction, and the rapid testing of cellular efficacy and toxicity of future novel therapeutic compounds.

Structural Design, Synthesis and Antioxidant, Antileishmania, Anti-Inflammatory and Anticancer Activities of a Novel Quercetin Acetylated Derivative.

Quercetin (Q) is a bioflavonoid with biological potential; however, poor solubility in water, extensive enzymatic metabolism and a reduced bioavailability limit its biopharmacological use. The aim of this study was to perform structural modification in Q by acetylation, thus, obtaining the quercetin pentaacetate (Q5) analogue, in order to investigate the biological potentials (antioxidant, antileishmania, anti-inflammatory and cytotoxicity activities) in cell cultures. Q5 was characterized by FTIR, 1H and 13C NMR spectra. The antioxidant potential was evaluated against the radical ABTS•+. The anti-inflammatory potential was evaluated by measuring the pro-inflammatory cytokine tumor necrosis factor (TNF) and the production of nitric oxide (NO) in peritoneal macrophages from BALB/c mice. Cytotoxicity tests were performed using the AlamarBlue method in cancer cells HepG2 (human hepatocarcinoma), HL-60 (promyelocytic leukemia) and MCR-5 (healthy human lung fibroblasts) as well as the MTT method for C6 cell cultures (rat glioma). Q and Q5 showed antioxidant activity of 29% and 18%, respectively, which is justified by the replacement of hydroxyls by acetyl groups. Q and Q5 showed concentration-dependent reductions in NO and TNF production (p < 0.05); Q and Q5 showed higher activity at concentrations > 40µM when compared to dexamethasone (20 µM). For the HL-60 lineage, Q5 demonstrated selectivity, inducing death in cancer cells, when compared to the healthy cell line MRC-5 (IC50 > 80 µM). Finally, the cytotoxic superiority of Q5 was verified (IC50 = 11 µM), which, at 50 µM for 24 h, induced changes in the morphology of C6 glioma cells characterized by a round body shape (not yet reported in the literature). The analogue Q5 had potential biological effects and may be promising for further investigations against other cell cultures, particularly neural ones.

Identification and characterization of potent, selective, and efficacious inhibitors of human arylamine N-acetyltransferase 1.

Arylamine N-acetyltransferase 1 (NAT1) plays a pivotal role in the metabolism of carcinogens and is a drug target for cancer prevention and/or treatment. A protein-ligand virtual screening of 2 million chemicals was ranked for predicted binding affinity towards the inhibition of human NAT1. Sixty of the five hundred top-ranked compounds were tested experimentally for inhibition of recombinant human NAT1 and N-acetyltransferase 2 (NAT2). The most promising compound 9,10-dihydro-9,10-dioxo-1,2-anthracenediyl diethyl ester (compound 10) was found to be a potent and selective NAT1 inhibitor with an in vitro IC50 of 0.75µM. Two structural analogs of this compound were selective but less potent for inhibition of NAT1 whereas a third structural analog 1,2-dihydroxyanthraquinone (a compound 10 hydrolysis product also known as Alizarin) showed comparable potency and efficacy for human NAT1 inhibition. Compound 10 inhibited N-acetylation of the arylamine carcinogen 4-aminobiphenyl (ABP) both in vitro and in DNA repair-deficient Chinese hamster ovary (CHO) cells in situ stably expressing human NAT1 and CYP1A1. Compound 10 and Alizarin effectively inhibited NAT1 in cryopreserved human hepatocytes whereas inhibition of NAT2 was not observed. Compound 10 caused concentration-dependent reductions in DNA adduct formation and DNA double-strand breaks following metabolism of aromatic amine carcinogens beta-naphthylamine and/or ABP in CHO cells. Compound 10 inhibited proliferation and invasion in human breast cancer cells and showed selectivity towards tumorigenic versus non-tumorigenic cells. In conclusion, our study identifies potent, selective, and efficacious inhibitors of human NAT1. Alizarin's ability to inhibit NAT1 could reduce breast cancer metastasis particularly to bone.

Effects of Lamotrigine and Topiramate on Glial Properties in an Astrocyte-Microglia Co-Culture Model of Inflammation.

BackgroundAstrocytes and microglia are involved in the pathophysiology of epilepsy and bipolar disorder with a link to inflammation. We aimed to investigate the effects of the antiepileptic and mood-stabilizing drugs lamotrigine (LTG) and topiramate (TPM) on glial viability, microglial activation, cytokine release, and expression of gap-junctional protein connexin 43 (Cx43) in different set-ups of an in vitro astrocyte-microglia co-culture model of inflammation.MethodsPrimary rat co-cultures of astrocytes containing 5% (M5, representing “physiological” conditions) or 30% (M30, representing “pathological, inflammatory” conditions) of microglia were treated with different concentrations of LTG and TPM for 24 hours. An 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to measure the glial cell viability. The microglial activation state was analyzed by immunocytochemistry. The pro-inflammatory tumor necrosis factor-α (TNF-α) and anti-inflammatory transforming growth factor-ß1 (TGF-ß1) cytokine levels were measured by enzyme-linked immunosorbent assay. The astroglial Cx43 expression was quantified by western blot.ResultsA significant reduction of the glial cell viability after incubation with LTG or TPM was observed in a concentration-dependent manner under all conditions. LTG caused no significant alterations of the microglial phenotypes. Under pathological conditions, TPM led to a significant concentration-dependent reduction of microglial activation. This correlated with increased astroglial Cx43 expression. TNF-α levels were not affected by LTG and TPM. Treatment with higher concentrations of LTG, but not with TPM, led to a significant increase in TGF-ß1 levels in M5 and M30 co-cultures.ConclusionsDespite the possible glial toxicity of LTG and TPM, both drugs reduced inflammatory activity, suggesting potential positive effects on the neuroinflammatory components of the pathogenesis of epilepsy and bipolar disorder.

Pseudoalteromone A, a Ubiquinone Derivative from Marine Pseudoalteromonas spp., Suppresses Melanogenesis.

An ubiquinone derivative, pseudoalteromone A (1), has been isolated from two marine-derived Pseudoalteromonas spp., APmarine002 and ROA-050, and its anti-melanogenesis activity was investigated. The anti-melanogenic capacity of pseudoalteromone A was demonstrated by assessing the intracellular and extracellular melanin content and cellular tyrosinase activity in the B16 cell line, Melan-a mouse melanocyte cell line, and MNT-1 human malignant melanoma cell line. Treatment with pseudoalteromone A (40 μg/mL) for 72 h reduced α-melanocyte-stimulating hormone (α-MSH)-induced intracellular melanin production by up to 44.68% in B16 cells and 38.24% in MNT-1 cells. Notably, pseudoalteromone A induced a concentration-dependent reduction in cellular tyrosinase activity in B16 cell, and Western blot analyses showed that this inhibitory activity was associated with a significant decrease in protein levels of tyrosinase and tyrosinase-related protein 1 (Tyrp-1), suggesting that pseudoalteromone A exerts its anti-melanogenesis activity through effects on melanogenic genes. We further evaluated the skin-whitening effect of pseudoalteromone A in the three-dimensional (3D) pigmented-epidermis model, MelanoDerm, and visualized the 3D distribution of melanin by two-photon excited fluorescence imaging in this human skin equivalent. Collectively, our findings suggest that pseudoalteromone A inhibits tyrosinase activity and expression and that this accounts for its anti-melanogenic effects in melanocytes.

NSAIDs Naproxen, Ibuprofen, Salicylate, and Aspirin Inhibit TRPM7 Channels by Cytosolic Acidification.

Non-steroidal anti-inflammatory drugs (NSAIDs) are used for relieving pain and inflammation accompanying numerous disease states. The primary therapeutic mechanism of these widely used drugs is the inhibition of cyclooxygenase 1 and 2 (COX1, 2) enzymes that catalyze the conversion of arachidonic acid into prostaglandins. At higher doses, NSAIDs are used for prevention of certain types of cancer and as experimental treatments for Alzheimer’s disease. In the immune system, various NSAIDs have been reported to influence neutrophil function and lymphocyte proliferation, and affect ion channels and cellular calcium homeostasis. Transient receptor potential melastatin 7 (TRPM7) cation channels are highly expressed in T lymphocytes and are inhibited by Mg2+, acidic pH, and polyamines. Here, we report a novel effect of naproxen, ibuprofen, salicylate, and acetylsalicylate on TRPM7. At concentrations of 3–30mM, they reversibly inhibited TRPM7 channel currents. By measuring intracellular pH with the ratiometric indicator BCECF, we found that at 300μM to 30mM, these NSAIDs reversibly acidified the cytoplasm in a concentration-dependent manner, and propose that TRPM7 channel inhibition is a consequence of cytosolic acidification, rather than direct. NSAID inhibition of TRPM7 channels was slow, voltage-independent, and displayed use-dependence, increasing in potency upon repeated drug applications. The extent of channel inhibition by salicylate strongly depended on cellular PI(4,5)P2 levels, as revealed when this phospholipid was depleted with voltage-sensitive lipid phosphatase (VSP). Salicylate inhibited heterologously expressed wildtype TRPM7 channels but not the S1107R variant, which is insensitive to cytosolic pH, Mg2+, and PI(4,5)P2 depletion. NSAID-induced acidification was also observed in Schneider 2 cells from Drosophila, an organism that lacks orthologous COX genes, suggesting that this effect is unrelated to COX enzyme activity. A 24-h exposure to 300μM–10mM naproxen resulted in a concentration-dependent reduction in cell viability. In addition to TRPM7, the described NSAID effect would be expected to apply to other ion channels and transporters sensitive to intracellular pH.

Toxicity of short multi-walled carbon nanotubes in Caenorhabditis elegans

Carbon nanotubes have become widely used due to their superior physiochemical properties, and so knowledge of their toxicity has become important. In this study, the toxic effects of nanotubes on Caenorhabditis elegans have been investigated. Multi-walled carbon nanotubes were characterized by transmission electron microscopy and particle size analysis. Upon exposure to the nanotubes at concentrations of 100 to 500 mg/L for 3 to 15 days, numbers and body lengths of the nematodes were not altered, but body thickness was raised and body movements were slightly decreased. A new type of toxicity-reproduction assay was performed to see the effect of multi-walled carbon nanotubes. The circular-form aggregation of starved larvae was counted. The number of aggregations was reduced with the increasing nanotube concentration by ∼27%. The concentration-dependent reduction was confirmed by counting the eggs in a single worm which demonstrated the negative effect of multi-walled carbon nanotubes on reproduction when the nematodes were subjected to prolonged exposure.

Actions of Camptothecin Derivatives on Larvae and Adults of the Arboviral Vector Aedes aegypti.

Mosquito-borne viruses including dengue, Zika, and Chikungunya viruses, and parasites such as malaria and Onchocerca volvulus endanger health and economic security around the globe, and emerging mosquito-borne pathogens have pandemic potential. However, the rapid spread of insecticide resistance threatens our ability to control mosquito vectors. Larvae of Aedes aegypti were screened with the Medicines for Malaria Venture Pandemic Response Box, an open-source compound library, using INVAPP, an invertebrate automated phenotyping platform suited to high-throughput chemical screening of larval motility. We identified rubitecan (a synthetic derivative of camptothecin) as a hit compound that reduced A. aegypti larval motility. Both rubitecan and camptothecin displayed concentration dependent reduction in larval motility with estimated EC50 of 25.5 ± 5.0 µM and 22.3 ± 5.4 µM, respectively. We extended our investigation to adult mosquitoes and found that camptothecin increased lethality when delivered in a blood meal to A. aegypti adults at 100 µM and 10 µM, and completely blocked egg laying when fed at 100 µM. Camptothecin and its derivatives are inhibitors of topoisomerase I, have known activity against several agricultural pests, and are also approved for the treatment of several cancers. Crucially, they can inhibit Zika virus replication in human cells, so there is potential for dual targeting of both the vector and an important arbovirus that it carries.

Beneficial actions of esculentin-2CHa(GA30) on high sucrose-induced oxidative stress in Drosophila melanogaster

Hyperglycaemia-induced oxidative stress plays a critical role in the development of diabetes and its complications. This study investigated actions of esculentin-2CHa(GA30) on high sucrose-induced oxidative stress in adult Drosophila melanogaster. Adult flies were exposed to diets containing graded concentrations of sucrose in the presence or absence of esculentin-2CHa(GA30) (5.0–10 μmol/kg diet) for 7 days. Effects of high sucrose diet and/or esculentin-2CHa(GA30) on survival and longevity of flies, and markers of oxidative stress, antioxidant status and glucose were assessed. High-sucrose diet (15–30%) and esculentin-2CHa(GA30) (5–10 μmol/kg diet) enhanced the percentage of surviving flies by 33.5%–46.2% (P < 0.01) and 7.4%–26.9% (P < 0.01) respectively. Concentration-dependent reduction in total thiol (19.3–51.3%, P < 0.01), reduced glutathione (22.6–54.9%, P < 0.05–0.01), catalase activity (36.8–57.3%, P < 0.05–0.01) and elevated glucose concentration (1.8–2.9-fold, P < 0.001) were observed in high sucrose-fed flies. Esculentin-2CHa(GA30) alone did not affect levels of total thiol, reduced glutathione, glucose and catalase activity. Improved survival (1.2–1.3-fold, P < 0.05–0.01) and longevity (1.3-fold) were observed in flies treated with the peptide (5.0 and 7.5 μmol/kg diet). Feeding on sucrose and esculentin-2CHa(1–30) (5.0 and 7.0 μmol/kg diet) for 7 days increased total thiol (2 - 3-fold, P < 0.001) and reduced glutathione (1.6–1.8-fold, P < 0.05) levels. Reduced catalase activity (21.4–36.4%, P < 0.01) and reduced glucose level (38.6–49.4%, P < 0.01) were observed in peptide-treated flies. Esculentin-2CHa(1–30) inhibited sucrose-induced generation of hydrogen peroxide (7.5–13.7%, P < 0.05) and nitric oxide (22.3–42.9%, P < 0.01) in adult flies. Overall, findings from this study offered further insights into the anti-oxidative properties of esculentin-2CHa(GA30).

Cyto-genotoxic potential of petroleum refinery wastewater mixed with domestic sewage used for irrigation of food crops in the vicinity of an oil refinery

Petroleum refinery wastewater combined with domestic sewage were collected from the open channel in the vicinity of Mathura oil refinery, UP (India) and analysed by inductively coupled plasma optical emission spectrometry (ICP-OES) and gas chromatography-mass spectrometry (GC-MS) for elemental analysis and organic pollutants, respectively. Several potentially toxic and non-toxic elements were found to be present in the wastewater samples. GC-MS analysis revealed the presence of several organic contaminants including pesticides. Wastewater samples were extracted using amberlite XAD4/8 resins and liquid-liquid extraction procedures using different organic solvents. The extracts were tested for their cyto-genotoxic potential using bacterial (Salmonella mutagenicity test, E. coli K-12 DNA repair defective mutants, Bacteriophage λ assay) and plant (Vigna mungo phytotoxicity test, Allium cepa chromosomal aberration assay) systems. A significant increase was observed in the number of revertants of TA97a, TA98 and TA100 strains with the test samples and XAD concentrated samples were found to be more mutagenic than liquid-liquid extracts. Colony forming units (CFUs) of DNA repair defective mutants of E. coli K-12 recA, lexA and polA declined significantly as compared to their isogenic wild-type counterparts with the test samples. Significant reduction in plaque forming units (PFUs) of bacteriophage λ was also found on treatment with the solvent extracts. Presence of several toxic pollutants in the wastewater apply prohibitive action on the seed germination process. Germination rate of Vigna mungo seeds as well as radicle and plumule lengths were found to be affected when treated with different concentration of wastewater as compared to control. Present study also indicated concentration dependent reduction in mitotic index of A. cepa i.e., 16.38% at 5% and 9.74% at 100% wastewater and percentage of aberrant cells were highest at 100% effluent. Present findings indicated that mutagenicity/genotoxicity of wastewater is due to the mixture of genotoxins; poses serious hazards to the receiving waterbodies which require continuous monitoring and remedial measures for their improvement.

Synergistic Antifungal Activity of Chito-Oligosaccharides and Commercial Antifungals on Biofilms of Clinical Candida Isolates.

The development of yeast biofilms is a major problem due to their increased antifungal resistance, which leads to persistent infections with severe clinical implications. The high antifungal activity of well-characterized chitosan polymers makes them potential alternatives for treating yeast biofilms. The activity of a chito-oligosaccharide with a depolymerization degree (DPn) of 32 (C32) and a fraction of acetylation (FA) of 0.15 on Candida sp. biofilms was studied. The results showed a concentration-dependent reduction in the number of viable cells present in C. albicans, C. glabrata, and C. guillermondii preformed biofilms in the presence of C32, especially on intermediate and mature biofilms. A significant decrease in the metabolic activity of yeast biofilms treated with C32 was also observed. The antifungals fluconazole (Flu) and miconazole (Mcz) decreased the number of viable cells in preformed early biofilms, but not in the intermediate or mature biofilms. Contrary to Flu or Mcz, C32 also reduced the formation of new biofilms. Interestingly, a synergistic effect on yeast biofilm was observed when C32 and Flu/Mcz were used in combination. C32 has the potential to become an alternative therapeutic agent against Candida biofilms alone or in combination with antifungal drugs and this will reduce the use of antifungals and decrease antifungal resistance.

Capsule Independent Antimicrobial Activity Induced by Nanochitosan against Streptococcus pneumoniae.

Background: Streptococcus pneumoniae remains a major cause of community-acquired pneumonia, meningitis, and other diseases, contributing significantly to high morbidity and mortality worldwide. Although it responds to antibiotics, their use is becoming limited due to the rise in antibiotic resistance, which necessitates the development of new therapeutics. Nanotechnology is used to counteract antimicrobial resistance. In this regard, polymeric nanoparticles (NPs) made of natural, biodegradable, biocompatible, and cationic polymers such as Chitosan (CNPs) exhibit wide-spectrum antimicrobial activity. Therefore, this study aimed to prepare CNPs, characterize their physiochemical characteristics: particle size (PZ), polydispersity index (PDI), and zeta potential (ZP), and investigate their antimicrobial activity against Streptococcus pneumoniae TIGR4 (virulent serotype 4) and its capsular mutant (∆cps). Methods: CNPs were prepared at 1, 2.5, and 5 mg/mL concentrations using the ion gelation method. Then, PZ, PDI, and ZP were characterized using a Zetasizer. Transmission electron microscopy (TEM) was used to visualize the CNP’s morphology. Broth and agar dilution methods were used to assess their antimicrobial activity. Cytotoxicity of prepared NPs on A549 cells and their effect on pneumococcal hemolysis were also investigated. Results: Spherical CNPs were produced with PZ ranging from 133.3 nm ± 0.57 to 423 nm ± 12.93 PDI < 0.35, and ZP from 19 ± 0.115 to 27 ± 0.819. The prepared CNPs exhibited antibacterial activity against TIGR4 and its capsule mutant with a minimum inhibitory concentration (MIC90) of 0.5 to 2.5 mg/mL in a non-acidic environment. The hemolysis assay results revealed that CNPs reduced bacterial hemolysis in a concentration-dependent manner. Their mammalian cytotoxicity results indicated that CNPs formed from low concentrations of Chitosan (Cs) were cytocompatible. Conclusion: Nanochitosan particles showed anti-pneumococcal activity regardless of the presence of capsules. They resulted in a concentration-dependent reduction in bacterial hemolysis and were cytocompatible at a lower concentration of Cs. These findings highlight the potential of CNPs in the treatment of pneumococcal diseases.

Adamantane carboxylic acids demonstrate mitochondrial toxicity consistent with oil sands-derived naphthenic acids

Naphthenic acids (NAs) are thought to be a primary cause of toxicity of oil sands process-affected water (OSPW). The purpose of this study was to determine if commercially available adamantane carboxylic acids act by mitrochondrial mechanisms similar to NAs found in OSPW. Mitochondria isolated from rainbow trout (Oncorhynchus mykiss) liver were exposed to commercially available adamantane acids, 3-hydroxyadamantane-1-carboxylic acid (CAS 42711-75-1) and 3,5-dimethyladamantane-1-carboxylic acid (CAS 14670-94-1), or to NAs extracted and purified from OSPW. The effects of these compounds on state 3 and 4 respiration, mitochondrial membrane potential, and mitochondrial reactive oxygen species (ROS; H2O2 production) were quantified. The compound 3-hydroxyadamantane-1-carboxylic acid only inhibited state 3 respiration at the highest concentration (2560 mg/L) and showed a concentration-dependent reduction in H2O2 production. Consistent with extracted OSPW NAs, 3,5-dimethyladamantane-1-carboxylic acid inhibited state 3 respiration and increased H2O2 production, but with a two-fold greater EC50 than the NA mixture extracted from OSPW. All three compounds uncoupled mitochondrial membrane potential and increased state 4 respiration. Based on these results, the adamantane 3,5-dimethyladamantane-1-carboxylic NAs may act via similar mitrochondrial mechanisms as NAs extracted from OSPW and could be used to further explore toxic mechanisms.

Ziziphus jujuba Mill. leaf extract restrains adipogenesis by targeting PI3K/AKT signaling pathway

The escalation in the global prevalence of obesity has focused attention on finding novel approaches for its management. Ziziphus jujuba Mill. (ZJL) leaf extract is reported as a traditional remedy for diverse pathological conditions, including obesity. The present study investigated whether ZJL affects adipogenic differentiation in human adipocytes. Additionally, following metabolite profiling of the extract, apigenin (APG), betulinic acid (BA) and maslinic acid (MA) were selected for biological activity evaluation. The possible interactions between APG, BA, MA and target proteins with a central role in adipogenesis were assessed through molecular docking. The potential mechanisms of ZJL, APG, BA and MA were identified using transcriptional analysis through real-time quantitative PCR and protein abundance evaluation by Western blotting. The obtained results revealed a concentration-dependent reduction of accumulated lipids after ZJL, BA and MA application. The key adipogenic transcription factors peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT-enhancer-binding protein alpha (C/EBPα) were strongly decreased at a protein level by all treatments. Moreover, the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway was found to be involved in the anti-adipogenic effect of ZJL, APG and BA. Collectively, our findings indicate that ZJL and its pure compounds hampered adipocyte differentiation through PI3K/AKT inhibition. Among the selected compounds, BA exhibits the most promising anti-adipogenic activity. Furthermore, being a complex mixture of phytochemicals, the ZJL extract could be utilized as source of yet unknown bioactive leads with potential implementation in obesity management.

The influence of alkaline treatment on the mechanical and structural properties of bacterial cellulose

The unique mechanical properties of hydrated bacterial cellulose make it suitable for biomedical applications. This study evaluates the effect of concentrated sodium hydroxide treatment on the structural and mechanical properties of bacterial cellulose hydrogels using rheological, tensile, and compression tests combined with mathematical modelling. Bacterial cellulose hydrogels show a concentration-dependent and irreversible reduction in shear moduli, compression, and tensile strength after alkaline treatment. Applying a poroelastic biphasic model to through-thickness compressive stress-relaxation tests showed the alkaline treatment to induce no significant change in axial compression, an effect was observed in the radial direction, potentially due to the escape of water from within the hydrogel. Scanning electron microscopy showed a more porous structure of bacterial cellulose. These results show how concentration-dependent alkaline treatment induces selective weakening of intramolecular interactions between cellulose fibres, allowing the opportunity to precisely tune the mechanical properties for specific biomedical application, e.g., faster-degradable materials.

Environmental modulators of diluted bitumen effects in juvenile pink salmon (Oncorhynchus gorbuscha)

Recent and potential expansions in the transportation of diluted bitumen (dilbit) through marine terminals in coastal regions of British Columbia require the examination of potential risks to estuarine species such as Pacific salmon. The estuarine habitat of out-migrated pink salmon (Oncorhynchus gorbuscha) exhibits dynamic temperature and salinity regimes, possibly modifying dilbit exposure, bioavailability and/or its effects. To examine dilbit toxicity and its modification by environmental stressors, juvenile pinks were subchronically exposed for 3 months to the water-accommodated fraction (WAF) of Cold Lake Blend dilbit (winter) in seawater at three salinities (7, 14, and 28‰ [temperature 12.5 °C]) and three temperatures (8.5, 12.5, and 16.5 °C [salinity of 28‰]). Temperature and salinity alone did not affect any measured endpoints in control fish. Dilbit exposure induced higher mortality at high (16.5 °C) and low temperatures (8.5 °C) as well as at higher salinity (28‰) in fish exposed to the highest dilution of WAF [total polycyclic aromatic compounds (TPAC) = 128.9 μg/L]. A concentration-dependent reduction of growth was evident in fish exposed to the medium (TPAC = 97.3 μg/L) and high dilution of WAF at higher temperatures (12.5 and 16.5 °C) and high salinity (28‰). At 28‰, swimming performance (Uburst) was decreased in fish exposed to the highest concentration of dilbit at all 3 temperatures. Gill Na+-K+-ATPase activity, white muscle lactate, glycogen, and triglyceride concentrations were altered by dilbit exposure and modified by temperature and salinity. In addition, gene expression associated with phase I biotransformation, energy metabolism, mitochondrial activity, and inflammation showed significant upregulation with exposure and temperature stress. Dilbit exposure at PAC concentrations in the ppb range, affected pink salmon at the molecular, biochemical, and whole organism level; effects that were exacerbated by environmental temperature and salinity.

Enhanced Apoptosis by Functionalized Highly Reduced Graphene Oxide and Gold Nanocomposites in MCF-7 Breast Cancer Cells.

Graphene nanocomposites have gained significant interest in a variety of biological applications due to their unique properties. Herein, we have studied the apoptosis-inducing ability and anticancer properties of functionalized highly reduced graphene oxide (HRG) and gold nanoparticles (Au NPs)-based nanocomposites (AP-HRG-Au). Samples were prepared under facile conditions via simple stirring and ultrasonication. All the samples were tested for their anticancer properties against different human cancer cell lines including lung (A549), liver (HepG2), and breast (MCF-7) cancer cells using doxorubicin as a positive control. In order to enhance the solubility and bioavailability of the sample, HRG was functionalized with 1-aminopyrene (1-AP) as a stabilizing ligand. The ligand also facilitated the homogeneous growth of Au NPs on the surface of HRG by offering chemically specific binding sites. The synthesis of nanocomposites and the surface functionalization of HRG were confirmed by UV–Vis, powder X-ray diffraction, and Fourier transform infrared spectroscopy. The structure and morphology of the as-prepared nanocomposites were established by high-resolution transmission electron microscopy. Because of the functionalization, the AP-HRG-Au nanocomposite exhibited enhanced physical stability and high dispersibility. A comparative anticancer study of pristine HRG, nonfunctionalized HRG-Au, and 1-AP-functionalized AP-HRG-Au nanocomposites revealed the enhanced apoptosis ability of functionalized nanocomposites compared to the nonfunctionalized sample, whereas the pristine HRG did not show any anticancer ability against all tested cell lines. Both HRG-Au and AP-HRG-Au have induced a concentration-dependent reduction in cell viability in all tested cell lines after 48 h of exposure, with a significantly higher response in MCF-7 cells compared to the remaining cells. Therefore, MCF-7 cells were selected to perform detailed investigations using apoptosis assay, cell cycle analysis, and reactive oxygen species measurements. These results suggest that AP-HRG-Au induces enhanced apoptosis in human breast cancer cells.