Estimated exposure to microplastics through national and local brands of bottled water in Central India.

Microplastics (MPs) are ubiquitous pollutants that affect various environmental matrices, including air, water, soil, food, and beverages. In India, there is limited research on microplastics in bottled drinking water, which is a significant route of MP exposure to the human body. To date, the data on the occurrence of MPs in national and local bottled water brands have not been studied and compared. Therefore, the current study focuses on the contamination of MPs in bottled water from different national and local brands procured from the market of Nagpur, India. The MPs were observed in all the analyzed samples. It was observed that the local bottled water showed higher MP contamination compared to national bottled water, with MP concentrations of 212 ± 100 MPs/L and 72 ± 36 MPs/L, respectively. The MPs were identified and characterized using microscopic and attenuated total reflectance-fourier transform infrared spectroscopy (ATR-FTIR) analysis, revealing that the dominant MP particles were fragments (71%), followed by fibers (23%), and others (6%). Among the observed particles, 50% of particles were black colored, followed by transparent (16%), red (13%), orange (8%), green (3%), blue (5%), and yellow (5%). The predominant polymer types were polyethylene (PE) and polyethylene terephthalate (PET). Overall, the pollution load indices suggested a moderate level of contamination in bottled water samples. Furthermore, the estimated annual human exposure to MPs was calculated as 5186 ± 3751 p/kg-bw/year for children and 1482 ± 1072 p/kg-bw/year for adults, making it a significant route of human exposure to MPs.

Strategy for Remediation of Polychlorinated Biphenyls-Contaminated Soil Through Redox Management Based on Electronegativity of the Contaminants.

Literature review reveals that Persistent Organic Pollutants (POPs), such as polychlorinated biphenyls (PCBs), are electron deficient compounds due to the presence of highly electronegative groups. Hence, they are more amenable to anaerobic biodegradation rather than oxidative metabolism. However, the studies on PCBs bioremediation are more inclined towards aerobic treatment. Besides, the past studies are mainly centered on screening and application of PCB-degrading microorganisms. In our opinion the degradative capacity is already present in the native microflora, and choice of electron donor is of paramount importance for faster reductive metabolism of PCBs. In this study, the use of methanol as electron donor with cow dung as the general microbial inoculum resulted in high specific rate of degradation (0.0542-0.0637 /day) for high-chlorinated biphenyls. The % removal of PCBs ranged between 67.7 and 71.7%. It may be the first study on the application of methanol as a cheap electron donor for PCBs biodegradation without bioaugmentation with specifically selected microorganisms.

Potential of Antarctic lipase from Acinetobacter johnsonii Ant12 for treatment of lipid-rich wastewater: screening, production, properties and applications.

The present study aimed to screen and optimize lipase production by the Antarctic strain Acinetobacter johnsonii Ant12 for lipid-rich wastewater treatment. Lipase production was successfully enhanced threefold through optimization of culture conditions. The optimum crude lipase activity was observed at 50 °C with high stability in a wide temperature range. The lipase also exhibited high activity and stability in the presence of solvents, metal ions, and surfactants. The crude lipase was used for the treatment of lipid-rich wastewater, which poses a significant challenge, as traditional removal methods are often inefficient or non-eco-friendly. In this study, bioaugmentation with Ant12 resulted in substantial lipid reduction in synthetic as well as real-world wastewater. Multiple linear regression analysis showed that lipid concentration and time were the most significant factors influencing lipid degradation. Bioaugmentation of real-world wastewater with Ant12 cells resulted in 84% removal of lipids in 72 h, while its crude lipase degraded 73.7% of lipids after 24 h. Thus, the specific rate of lipid degradation was higher for crude lipase (0.095/h) than the whole cell treatment (0.031/h). Economic analysis revealed that crude lipase production was much cheaper, faster and more eco-friendly than purified or partially purified lipase production, which justifies its use in wastewater treatment. The high activity of enzyme also implicates its application as a detergent additive. In our knowledge, it is the first study to establish A. johnsonii isolate from Antarctica for lipid-rich wastewater treatment.

Screening, identification, and characterization of lipase-producing halotolerant Bacillus altitudinis Ant19 from Antarctic soil.

Potent lipase-producing and halotolerant Bacillus altitudinis Ant19 strain was screened and isolated from Antarctic soil. The isolate showed broad-range lipase activity against different lipid substrates. Presence of lipase activity was confirmed by PCR amplification and sequencing of the lipase gene from Ant19. The study attempted to establish the use of crude extracellular lipase extract as cheap alternative to purified enzyme by characterizing the crude lipase activity and testing it in certain practical applications. Crude lipase extract from Ant19 showed high stability at 5-28 â„ƒ (> 97%), while lipase activity was noted in a wide temperature range of 20-60 â„ƒ (> 69%), with optimum activity at 40 â„ƒ (117.6%). The optimum lipolytic activity was noted at pH 8 with good activity and stability in alkaline conditions (pH 7-10). Moreover, the lipase activity was substantially stable in various solvents, commercial detergents, and surfactants. It retained 97.4% activity in 1% solution of commercial Nirma detergent. Besides, it was non-regiospecific, and active against substrates having different fatty acid chain lengths with preference for shorter chain length. Further, the crude lipase enhanced the oil stain removal efficiency of commercial detergent from 52 to 77.9%, while 66% oil stain was removed using crude lipase alone. Immobilization process improved the storage stability of crude lipase for 90 days. In our knowledge, it is the first study on characterization of lipase activity from B. altitudinis, which has promising applications in various fields.

Identification of environmental stress parameters to study the natural colour change of water in highly saline inland Crater Lake at Lonar, India.

Lonar Lake is a highly saline inland water body created by a crater in Maharashtra, India. A rare occurrence of the colour change of lake water from green to brown and eventually to pinkish-red was observed in Lonar in June 2020. This phenomenon attracted the attention of researchers, academicians and interestingly legal fraternity to understand the causes of colour change. The literature studies coupled the phenomenon of colouration of water to three aspects: the presence of halophilic Halobacterium salinarum or an algal species of Dunaliella (Dunaliella salina) or oxidization of metals (Fe and Mn) present in water. A comprehensive study was done to understand and assess the change in the colour of Lonar Lake water. The green colour of the lake is primarily due to the dominance of chlorophyll-a pigment in the algae population. The stressed condition in June 2020 adversely affected the photosynthesis activity of Dunaliella sp. resulting in the red colouration of the species. This red colour of Dunaliella sp. is due to the formation of a pigment named carotenoid which is similar to that in halophilic bacteria. This pigment completely hides the green chloroplast, and water turns pinkish-red. This study describes detailed investigations of environmental and climatic parameters to determine possible causes of abiotic stress on the algae population of the lake. The major factors contributing to the stressed conditions are high dissolved solids, alkalinity and alkaline pH due to salts in the lake water due to evaporation losses and limited rainfall over the months. The study further verified whether the colour change is a cyclic event and predicted possible lake conditions for the event of colour change to occur in the future.

Characterization and removal of microplastics in a sewage treatment plant from urban Nagpur, India.

Sewage treatment plant (STP) acts as a potential source of microplastic contamination in the environment. The presence of microplastics in the sewage treatment plant is reported over the globe in varying concentrations. Hence, the current study is intended to evaluate the presence and abundance of microplastics occurring in sewage treatment plants in India. The samples were processed through digestion and density separation, followed by microscopic and polymer identification through Fourier transform infrared spectroscopy. Also, different wastewater parameters were studied to analyze their influence. High microplastic concentrations were detected in the influent (1860 ± 265 MPs/L), which reduced by > 90%, to around 148 ± 51 MPs/L in the effluent. The concentration of microplastics in sewage sludge was 830 MPs/kg. The prominent plastic types identified include low-density polyethylene, polypropylene, polyurethane, polyvinyl chloride, and rayon. The smaller particles prevail in the effluent, releasing around 30 billion particles per day to the environment. This suggests that the current STP is efficient in removing the majority of the particles, but considerations are needed to avoid the ecological risks associated.

Assessment of occupational exposure to diesel particulate matter through evaluation of 1-nitropyrene and 1-aminopyrene in surface coal miners, India.

DPM (diesel particulate matter) is ubiquitously present in the mining environment and is known for mutagenicity and carcinogenicity to humans. However, its health effects in surface coal mines are not well studied, particularly in India. In this study, DPM exposure and corresponding exposure biomarkers were investigated in four different surface coal mines in Central India. To document and evaluate the DPM exposure in surface coal miners, we characterized 1-NP (1-nitropyrene) in the mining environment as surrogate for DPM using Sioutas Cascade Impactor. Exposure biomarkers were analyzed by collecting post work shift (8-h work shift) urine samples and determining the concentrations of 1-aminopyrene (1-AP) as a metabolite of 1-NP and 8-hydroxydeoxyguanosine (8OHdG) as DNA damage marker. We observed high concentration of 1-NP (7.13-52.46 ng/m3) in all the mines compared with the earlier reported values. The average creatinine corrected 1-AP and 8OHdG levels ranged 0.07-0.43 [Formula: see text]g/g and 32.47-64.16 [Formula: see text]g/g, respectively, in different mines. We found 1-AP in majority of the mine workers' urine (55.53%) and its level was higher than that reported for general environmental exposure in earlier studies. Thus, the study finding indicates occupational exposure to DPM in all the four mines. However, the association between 1-NP level and exposure biomarkers (1-AP and 8OHdG) was inconsistent, which may be due to individual physiological variations. The data on exposure levels in this study will help to understand the epidemiological risk assessment of DPM in surface coal miners. Further biomonitoring and cohort study are needed to exactly quantify the occupational health impacts caused by DPM among coal miners.

Environmental prevalence, fate, impacts, and mitigation of microplastics-a critical review on present understanding and future research scope.

Microplastics are considered to be ubiquitous and widespread emerging contaminants. They are persistent in the nature and pose considerable harm to the environment. Their omnipresence is documented in almost all aquatic habitats, several atmospheric and terrestrial environments, and also in human consumables. The objective of this review is to provide an overview of the environmental prevalence of the microplastics in all environmental compartments, and their possible adverse impacts. It also presents review of the studies conducted in India and the epitome of potential mitigation measures. The need and direction of future research are highlighted. The review will help in determining the exposure levels, environmental consequences, and risk estimations, and will guide the researchers and policymakers.

Purification, characterization, and crystal structure of YhdA-type azoreductase from Bacillus velezensis.

Azoreductases are being extensively investigated for their ability to initiate degradation of recalcitrant azo dyes through reduction of azo bonds. There is great interest in studying their diversity, structure, and function to facilitate better understanding and effective application. Current study reports azoreductase enzyme from Bacillus velezensis, which showed 69.5% identity to the Bacillus subtilis azoreductase YhdA. The enzyme was homotetrameric and molecular weight of each subunit was 20 kDa. It decolorized azo dyes with different structures. The Vmax for decolorization of congo red, methyl orange and methyl red was 14.7, 28.6, and 77.9 nmol/min/mg, respectively. The enzyme contained FMN as cofactor and used NADPH as the favored co-substrate. It was oxygen-insensitive, but the presence of reducing agents enhanced its activity, which is a new finding. The azoreductase expression in B. velezensis was found to be unaffected by addition of azo dyes, although azo dyes are known to induce azoreductase expression in few organisms. The enzyme was thermostable with melting temperature of 89.5°C and functioned in wide temperature range. Further, the enzyme was crystallized and its structure was solved. The structural basis of its functional attributes is discussed. In our knowledge, this is the first report on characterization of azoreductase enzyme from B. velezensis.

Remediation of different nitroaromatic pollutants by a promising agent of Cupriavidus sp. strain a3.

Nitrobenzene, nitrotoluenes and nitrobenzoic acid are toxic and mutagenic. Their removal from the environment is necessary to avoid health and environmental damage. In this study, Cupriavidus strain a3 was found to utilize 2-nitrotoluene (2NT), 3-nitrotoluene (3NT), 4-nitrotoluene (4NT), nitrobenzene (NB) and 2-nitrobenzoic acid (2NBA) as carbon and nitrogen source, resulting in their detoxification. The metabolism involved reductive transformation of nitroaromatics to the corresponding amines followed by cleavage of amino group to release ammonia. Cell free extract showed nitroreductase activity in the range of 310-389 units/mg. NB was reduced to form benzamine and 4-aminophenol, 2NT was reduced to 2-aminotoluene, whereas 2NBA was reduced to form 2-aminobenzoic acid. Similarly, 3NT was metabolized to 3-aminotoluene and 2-amino-4-methylphenol, while 4NT was reduced to 4-nitrosotoluene and 4-aminotoluene. Cytotoxicity and apoptosis assays using Jurkat cell line, and Ames test were used to evaluate the detoxification of nitroaromatics during biodegradation. Biodegradation with Cupriavidus resulted in 2.6-11 fold increase in cell viability, 1.3-2.3 fold reduction in apoptosis, 1.6-55 fold reduction in caspase-3 activation, and complete disappearance of mutagenic activity. In soil microcosm, bioaugmentation with Cupriavidus resulted in 16-59% degradation of various nitroaromatics, as against <14% degradation without bioaugmentation. Thus, the present study reflects promising capability of Cupriavidus strain a3 in degradation and detoxification of multiple nitroaromatics.

Environmental persistence, hazard, and mitigation challenges of nitroaromatic compounds.

Nitroaromatic compounds (NACs) are extensively used in different industries and are synthesized in large quantity due to their heavy demand worldwide. The broad use of NACs poses a serious pollution threat. The treatment processes used for the removal of NACs are not effective and sustainable, leading to their release into the environment. The nitro group attached to benzene ring makes the compounds recalcitrant due to which they persist in the environment. Being hazardous to human as well as other living organisms, NACs are listed in the USEPA's priority pollutant group. This review provides updated information on the sources of NACs, prevalence in different environmental matrices, and recent developments in methods of their detection, with emphasis on current trends as well as future prospects. The harmful effects of NACs due to exposure through different routes are also highlighted. Further, the technologies reported for the treatment of NACs, including physico-chemical and biological methods, and the challenges faced for their effective implementation are discussed. Thus, the review discusses relevant issues in detail making suitable recommendations, which can be helpful in guiding further research in this subject.

Role of fluoride induced epigenetic alterations in the development of skeletal fluorosis.

Fluoride is an essential trace element required for proper bone and tooth development. Systemic high exposure to fluoride through environmental exposure (drinking water and food) may result in toxicity causing a disorder called fluorosis. In the present study, we investigated the alteration in DNA methylation profile with chronic exposure (30 days) to fluoride (8 mg/l) and its relevance in the development of fluorosis. Whole genome bisulfite sequencing (WGBS) was carried out in human osteosarcoma cells (HOS) exposed to fluoride. Whole genome bisulfite sequencing (WGBS) and functional annotation of differentially methylated genes indicate alterations in methylation status of genes involved in biological processes associated with bone development pathways. Combined analysis of promoter DNA hyper methylation, STRING: functional protein association networks and gene expression analysis revealed epigenetic alterations in BMP1, METAP2, MMP11 and BACH1 genes, which plays a role in the extracellular matrix disassembly, collagen catabolic/organization process, skeletal morphogenesis/development, ossification and osteoblast development. The present study shows that fluoride causes promoter DNA hypermethylation in BMP1, METAP2, MMP11 and BACH1 genes with subsequent down-regulation in their expression level (RNA level). The results implies that fluoride induced DNA hypermethylation of these genes may hamper extracellular matrix deposition, cartilage formation, angiogenesis, vascular system development and porosity of bone, thus promote skeletal fluorosis.

Simultaneous quantitative monitoring of four indicator contaminants of emerging concern (CEC) in different water sources of Central India using SPE/LC-(ESI)MS-MS.

Environmental occurrence of CECs poses a great threat to both aquatic life and human health. The aim of this study was to optimize and validate SPE/LC-(ESI)MS-MS method for simultaneous quantitative monitoring of two sub-classes of CECs (pharmaceuticals and hormones) and to estimate the concentrations of select CECs in environmental water samples. For all the tested analytes, recoveries in laboratory reagent water were greater than 81%. Average percent (relative standard deviation) RSD of the analytes in recovery, repeatability, and reproducibility experiments were ≤ 10%. Determination coefficients (r2) of primidone, diclofenac, testosterone, and progesterone were estimated to be 0.9979, 0.9972, 0.9968, and 0.9962, respectively. Limits of detection (LOD) for primidone, diclofenac, testosterone, and progesterone were 4.63 ng/L, 5.36 ng/L, 0.55 ng/L, and 0.88 ng/L, respectively. Limits of quantification (LOQ) for primidone, diclofenac, testosterone, and progesterone were 14.72 ng/L, 17.06 ng/L, 1.766 ng/L, and 2.813 ng/L, respectively. Average recoveries in environmental water and wastewater samples were greater than 74% and RSD were ≤ 7%. Trace levels (68.33-125.70 ng/L) of primidone were detected in four environmental water samples, whereas diclofenac was not detected in any of the tested sample. Trace levels of progesterone were observed in two environmental samples (16.64 -203.73 ng/L), whereas testosterone was detected in STP inlet sample (178.16 ng/L).

Molecular mechanism of apoptosis induction in Jurkat E6-1 cells by Tribulus terrestris alkaloids extract.

The present study demonstrates apoptosis-inducing potential and mechanism of action of Tribulus terristris alkaloid extract in Jurkat E6-1 cancer cell line. Liquid Chromatography-Mass Spectrometry and High Resolution-Mass Spectrometry analysis identified the presence of four N-feruloyltyramine derivatives, namely trans-N-feruloyl-3-hydroxytyramine (1), trans-N-coumaroyltyramine (2), trans-N-feruloyltyramine (3) and trans-N-feruloyl-3-ethoxytyramine (4) in the alkaloid extract. Compounds 2 and 3 have not been yet reported in the alkaloid extract of T. terristris. In silico analysis revealed therapeutic potential of N-feruloyltyramine derivatives and strong binding efficiency to both chains of Tumor Necrosis Factor Receptor 1. Treatment of alkaloids extract to Jurkat E6-1 clone induced dose-dependent cytotoxicity (LC50 140.4 µg mL-1). Jurkat cells treated with alkaloids extract at sub-lethal concentration showed DNA fragmentation, enhancement in caspase-3 activity and phosphatidylserine translocation (apoptosis indicator) compared to control cells. Gene expression analysis using Human Apoptosis RT2 Profiler PCR Array analysis upon alkaloid treatment was found to significantly alter expression of critical genes such as TNFR1, FADD, AIFM, CASP8, TP53, DFFA and NFKB1. These genes are predicted to mediate apoptotic cell death via both intrinsic and extrinsic apoptosis pathway. In summary, we report the identification of new N-feruloyltyramine derivatives from alkaloid extract of T. terristris fruit with probable anti-leukemic and pharmacological potential.

Role of fluoride induced histone trimethylation in development of skeletal fluorosis.

Chronic exposure to fluoride has been associated with the development of skeletal fluorosis. Limited reports are available on fluoride induced histone modification. However, the role of histone modification in the pathogenesis of skeletal fluorosis is not investigated. In the present study, we have investigated the role of fluoride induced histone modification on fluorosis development using human osteosarcoma (HOS) cell line. The expression of histone methyltransferases (EHMT1 and EHZ2) and level of global histone trimethylation (H3K9 and H3K27) have been assessed and observed to be increased significantly after fluoride exposure (8 mg/L). EpiTect chromatin immunoprecipitation (CHIP) qPCR Array (Human TGFß/BMP signaling pathway) was performed to assess the H3K9 trimethylation at promoter regions of pathway-specific genes. H3K9 ChIP PCR array analysis identified hyper H3K9 trimethylation in promoter regions of TGFBR2 and SMAD3. qPCR and STRING analysis was carried out to determine the repressive epigenetic effect of H3K9 trimethylation on expression pattern and functional association of identified genes. Identified genes (TGFBR2 and SMAD3) showed down-regulation which confirms the repressive epigenetic effect of promoter H3K9 hyper trimethylation. Expression of two other vital genes COL1A1 and MMP13 involved in TGFBR2-SMAD signaling pathway was also found to be down-regulated with a decrease in expression of TGFBR2 and SMAD3. STRING analysis revealed functional association and involvement of identified genes TGFBR2, SMAD3, COL1A1 and MMP13 in the collagen and cartilage development/morphogenesis, connective tissue formation, bio-mineral tissue development, endochondral bone formation, bone and skeletal morphogenesis. In conclusion, present investigation is a first attempt to link fluoride induced hyper H3K9 tri-methylation mediated repression of TGFBR2 and SMAD3 with the development of skeletal fluorosis.

Manganese exposure: Linking down-regulation of miRNA-7 and miRNA-433 with α-synuclein overexpression and risk of idiopathic Parkinson's disease.

Manganese is an essential trace element however elevated environmental and occupational exposure to this element has been correlated with neurotoxicity symptoms clinically identical to idiopathic Parkinson's disease. In the present study we chronically exposed human neuroblastoma SH-SY5Y cells to manganese (100µM) and carried out expression profiling of miRNAs known to modulate neuronal differentiation and neurodegeneration. The miRNA PCR array results reveal alterations in expression levels of miRNAs, which have previously been associated with the regulation of synaptic transmission and apoptosis. The expressions of miR-7 and miR-433 significantly reduced upon manganese exposure. By in silico homology analysis we identified SNCA and FGF-20as targets of miR-7 and miR-433. We demonstrate an inverse correlation in expression levels where reduction in these two miRNAs causes increases in SNCA and FGF-20. Transient transfection of SH-SY5Y cells with miR-7 and miR-433 mimics resulted in down regulation of SNCA and FGF-20 mRNA levels. Our study is the first to uncover the potential link between manganese exposure, altered miRNA expression and parkinsonism: manganese exposure causes overexpression of SNCA and FGF-20 by diminishing miR-7 and miR-433 levels. These miRNAs may be considered critical for protection from manganese induced neurotoxic mechanism and hence as potential therapeutic targets.

Structural and functional characterization of mercuric reductase from Lysinibacillus sphaericus strain G1.

In response to the widespread presence of inorganic Hg in the environment, bacteria have evolved resistance systems with mercuric reductase (MerA) as the key enzyme. MerA enzymes have still not been well characterized from gram positive bacteria. Current study reports physico-chemical, kinetic and structural characterization of MerA from a multiple heavy metal resistant strain of Lysinibacillus sphaericus, and discusses its implications in bioremediation application. The enzyme was homodimeric with subunit molecular weight of about 60 kDa. The Km and Vmax were found to be 32 µM of HgCl2 and 18 units/mg respectively. The enzyme activity was enhanced by ß-mercaptoethanol and NaCl up to concentrations of 500 µM and 100 mM respectively, followed by inhibition at higher concentrations. The enzyme showed maximum activity in the pH range of 7-7.5 and temperature range of 25-50 °C, with melting temperature of 67 °C. Cu2+ exhibited pronounced inhibition of the enzyme with mixed inhibition pattern. The enzyme contained FAD as the prosthetic group and used NADPH as the preferred electron donor, but it showed slight activity with NADH as well. Structural characterization was carried out by circular dichroism spectrophotometry and X-ray crystallography. X-ray confirmed the homodimeric structure of enzyme and gave an insight on the residues involved in catalytic binding. In conclusion, the investigated enzyme showed higher catalytic efficiency, temperature stability and salt tolerance as compared to MerA enzymes from other mesophiles. Therefore, it is proposed to be a promising candidate for Hg2+ bioremediation.

Application of cell-based assays for toxicity characterization of complex wastewater matrices: Possible applications in wastewater recycle and reuse.

Exposure to pre-concentrated inlet or outlet STP wastewater extracts at different concentrations (0.001% to 1%) induced dose-dependent toxicity in MCF-7 cells, whereas drinking water extracts did not induce cytotoxicity in cells treated. GC-MS analysis revealed the occurrence of xenobiotic compounds (Benzene, Phthalate, etc.) in inlet/outlet wastewater extracts. Cells exposed to inlet/outlet extract showed elevated levels of reactive oxygen species (ROS: inlet: 186.58%, p<0.05, outlet, 147.8%, p<0.01) and loss of mitochondrial membrane potential (Δψm: inlet, 74.91%, p<0.01; outlet, 86.70%, p<0.05) compared to the control. These concentrations induced DNA damage (Tail length: inlet: 34.4%, p<0.05, outlet, 26.7%, p<0.05) in treated cells compared to the control (Tail length: 7.5%). Cell cycle analysis displayed drastic reduction in the G1 phase in treated cells (inlet, G1:45.0%; outlet, G1:58.3%) compared to the control (G1:67.3%). Treated cells showed 45.18% and 28.0% apoptosis compared to the control (1.2%). Drinking water extracts did not show any significant alterations with respect to ROS, Δψm, DNA damage, cell cycle and apoptosis compared to the control. Genes involved in cell cycle and apoptosis were found to be differentially expressed in cells exposed to inlet/outlet extracts. Herein, we propose cell-based toxicity assays to evaluate the efficacies of wastewater treatment and recycling processes.

Oxidative Stress and Genotoxicity of Zinc Oxide Nanoparticles to Pseudomonas Species, Human Promyelocytic Leukemic (HL-60), and Blood Cells.

In the present study, toxicity of commercial zinc oxide nanoparticles (ZnO NPs) was studied on the bacterium Pseudomonas sp., human promyelocytic leukemia (HL-60) cells, and peripheral blood mononuclear cells (PBMC). The toxicity was assessed by measuring growth, cell viability, and protein expression in bacterial cell. The bacterial growth and viability decreased with increasing concentrations of ZnO NP. Three major proteins, ribosomal protein L1 and L9 along with alkyl hydroperoxides reductase, were upregulated by 1.5-, 1.7-, and 2.0-fold, respectively, after ZnO NP exposure. The results indicated oxidative stress as the leading cause of toxic effect in bacteria. In HL-60 cells, cytotoxic and genotoxic effects along with antioxidant enzyme activity and reactive oxygen species (ROS) generation were studied upon ZnO NP treatment. ZnO NP exhibited dose-dependent increase in cell death after 24-h exposure. The DNA-damaging potential of ZnO NP in HL-60 cells was maximum at 0.05 mg/L concentration. Comet assay showed 70-80% increase in tail DNA at 0.025 to 0.05 mg/L ZnO NP concentration. A significant increase of 1.6-, 1.4-, and 2.0-fold in ROS level was observed after 12 h. Genotoxic potential of ZnO NPs was also demonstrated in PBMC through DNA fragmentation. Thus, ZnO NP, besides being an essential element having antibacterial activity, also showed toxicity towards human cells (HL-60 and PBMC).

Fluoride-Induced Oxidative and Inflammatory Stress in Osteosarcoma Cells: Does It Affect Bone Development Pathway?

Oxidative stress is reported to negatively affect osteoblast cells. Present study reports oxidative and inflammatory signatures in fluoride-exposed human osteosarcoma (HOS) cells, and their possible association with the genes involved in osteoblastic differentiation and bone development pathways. HOS cells were challenged with sublethal concentration (8 mg/L) of sodium fluoride for 30 days and analyzed for transcriptomic expression. In total, 2632 transcripts associated with several biological processes were found to be differentially expressed. Specifically, genes involved in oxidative stress, inflammation, osteoblastic differentiation, and bone development pathways were found to be significantly altered. Variation in expression of key genes involved in the abovementioned pathways was validated through qPCR. Expression of serum amyloid A1 protein, a key regulator of stress and inflammatory pathways, was validated through western blot analysis. This study provides evidence that chronic oxidative and inflammatory stress may be associated with the fluoride-induced impediment in osteoblast differentiation and bone development.