Comprehensive antifungal investigation of green synthesized silver nanoformulation against four agriculturally significant fungi and its cytotoxic applications.

The present study reports the green synthesis of silver nanoparticles (AgNPs) in powder form using the leaf extract of Azadirachta indica. The synthesis of AgNPs was confirmed by UV-vis spectroscopy, FTIR, XRD, FESEM, and EDX. The synthesized AgNPs were in a powdered state and dispersed completely in 5% polyethylene glycol (PEG) and demonstrated prolonged shelf life and enhanced bioavailability over a year without any aggregation. The resulting silver nanoformulation demonstrated complete inhibition against Sclerotinia sclerotiorum and Colletotrichum falcatum and 68% to 80% inhibition against Colletotrichum gloeosporioides and Rhizoctonia solani respectively, at 2000 ppm. The EC50 values determined through a statistical analysis were 66.42, 157.7, 19.06, and 33.30 ppm for S. sclerotiorum, C. falcatum, C. gloeosporioides, and R. solani respectively. The silver nanoformulation also established significant cytotoxicity, with a 74.96% inhibition rate against the human glioblastoma cell line U87MG at 250 ppm. The IC50 value for the cancerous cell lines was determined to be 56.87 ppm through statistical analysis. The proposed silver nanoformulation may be used as a next-generation fungicide in crop improvement and may also find application in anticancer investigations. To the best of our knowledge, this is also the first report of silver nanoformulation demonstrating complete inhibition against the economically significant phytopathogen C. falcatum.

Combined Effects of Fluoride and Arsenic on Mitochondrial Function in the Liver of Rat.

Biochemical and/or molecular mechanisms of arsenic or fluoride toxicity in experimental animals have been widely investigated in the recent past. However, their combined effects on target cells/organelle are poorly understood. The present study was executed to delineate their combined effects on mitochondrial function in the liver of rat. Female Wistar rats (140 ± 20 g) were force fed individually or in combination with sodium arsenate (4 mg/kg body weight) and sodium fluoride (4 mg/kg body weight) for 90 days. Thereafter, established markers of mitochondrial function viz. mitochondrial lipid peroxidation, oxidative phosphorylation, ATPase, succinic dehydrogenase, and caspase-3 activity were determined. Cytochrome C release and oxidative DNA damage were also estimated in the liver of respective groups of rats. The study showed significant differences in these results amongst the three groups. Observations on parameters viz. LPO, cytochrome-C, caspase-3, and 8-OHdG suggested an antagonistic relationship between these two elements. Results on ATPase, SDH, and ADP:O ratio indicated synergism. It is concluded that AsIII + F in combination may express differential effects on signalling pathways and proapoptotic/antiapoptotic proteins/genes that contribute to liver cell death. Interaction of As and F with mitochondria.

Potential toxicity of nickel nano and microparticles on the reproductive system of female rats: A comparative time-dependent study.

Increased application of engineered nanoparticles in different sectors viz. agriculture, commerce, industry, and medicine has raised serious public health issues. Nanoparticles of nickel have been increasingly used as catalysts, conductive pastes, adhesives, nanowires, and nanofilters. Human and animal exposure to these particles may cause toxicity in different organs/systems. Studies made in the past had demonstrated their toxicity in liver, kidney, and lungs. However, their reproductive effects remain poorly understood. Therefore, the present study on reproductive toxicity of nickel nanoparticles (<30 nm) was executed in female Wistar rats. A comparison of results obtained in nickel microparticle-treated rats was also made. Rats were administered nano and microparticles through gavage at a dosage of 5 mg/kg body weight each for two exposure periods; that is, 15 and 30 days. Ovaries removed from these rats were analyzed to study the effects of nickel bioaccumulation on synthesis of steroid hormones, lipid peroxidation, apoptosis, and oxidative stress. Structural changes were monitored through histopathological and ultrastructural observations. The present study showed exposure time-dependent differences in the toxicity of nickel nano and microparticles in the ovary of rats. Nano nickel was cumulative in the ovaries. It affected steroidogenesis. Further, increased generation of reactive oxygen species and enhanced oxidative stress may have contributed to cytotoxicity. It was concluded that exposure to nano nickel might induce irreversible damage in the ovaries of rat.

Significance of Inflammation and Apoptosis in Hepatocellular Death in Rat, Co-treated with Arsenic and Fluoride.

Health effects elicited by combined environmental exposures to xenobiotics, in many instances, still remain unresolved. One of these examples is the combined toxicity of arsenic and fluoride. The present study was undertaken to delineate the role of inflammation and apoptosis in hepatocellular death caused by co-exposure to arsenic and fluoride in rat. Sodium arsenate (4 mg/kg body weight) and sodium fluoride (4 mg/kg body weight) were administered to female Wistar rats, individually and in combination, for 90 days. Results on tumor necrotic factor-α (TNF-α), interleukin-12 (IL-12), and comet assay showed increased values in comparison to those obtained in arsenic- or fluoride-treated rats. Results on NO, TBARS, and caspase-9 showed higher values than fluoride-treated rats but lower levels than arsenic-treated rats. It is hypothesized that increased generation of nitric oxide induces the release of cytokines that activates caspase-9. Caspase-9 promotes the synthesis of caspase-3 that executes apoptosis. Histopathological observations on apoptotic bodies and Kupffer cells support these observations.

Zinc Oxide Nanoparticles Ameliorate Dimethylnitrosamine-Induced Renal Toxicity in Rat.

Dimethylnitrosamine (DMN) is an established carcinogen. It is toxic to several organs, viz., the liver, kidney, and lungs, and immune system. Several drugs have been used in the past to modulate its toxicity using experimental animal models. The present study was designed to investigate the effect of zinc oxide nanoparticles (ZnONPs) on renal toxicity caused by DMN in laboratory rat. Since oxidative mechanisms are mainly involved in its toxicity, the proposed study focuses on the amelioration of oxidative stress response by ZnONPs, if any. The present results show that administration of ZnONPs (50 mg/kg body weight/rat) to DMN (2 µl/100 g body weight/rat)-treated rats diminuted the concentration of malonaldehyde, H2O2, and NO in the kidney. However, reduced glutathione (GSH) concentration increased after ZnONP treatment. Results on glutathione S-transferase and glutathione peroxidase favored its antioxidative effects. These results are supported by the recovery of oxidative DNA damage and less pronounced histopathological changes in the kidney. It is hypothesized that ZnONPs might be toxic to renal tissue; however, its strong therapeutic/antioxidative potential helps in ameliorating DMN-induced renal toxicity in rat.

Attributes of oxidative stress in the reproductive toxicity of nickel oxide nanoparticles in male rats.

The nanoparticles of nickel are now being widely used in industrial, commercial, and biomedical applications. In recent years, health safety issues posed by them have aroused concerns among health scientists. The aim of the present study was to investigate the role of oxidative stress in male reproductive toxicity induced by nickel oxide nanoparticles in rats. Male Wistar rats (140-170 g) were administered with nickel oxide nanoparticles (NiONPs) (particles size <30 nm) (5 mg/kg body weight) by gavage for 30 days. Its effects on different parameters, viz., sperm count, motility, and morphology, were investigated. DNA damage in sperms was monitored through comet assay. All these observations indicated a spermicidal effect of NiONPs. Results on lipid peroxidation (MDA, H2O2, and NO) and oxidative stress (GSH, GPx, and catalase) thus studied in testes exhibited adverse effects of NiONPs. Histopathological results on male reproductive organs, viz., testis, epididymis, vas deferens, seminal vesicles, and prostate also demonstrated moderate to severe toxicity. A comparison of these results with those obtained on nickel oxide microparticle (NiOMP)-treated rats showed that NiONPs are more toxic than NiOMPs. Furthermore, NiONPs could create an imbalance between oxidants and antioxidants in the testes. It is concluded that redox imbalance in testes constitutes a major mechanism of NiONP-induced reproductive toxicity.

Tumor reversion: a dream or a reality.

Reversion of tumor to a normal differentiated cell once considered a dream is now at the brink of becoming a reality. Different layers of molecules/events such as microRNAs, transcription factors, alternative RNA splicing, post-transcriptional, post-translational modifications, availability of proteomics, genomics editing tools, and chemical biology approaches gave hope to manipulation of cancer cells reversion to a normal cell phenotype as evidences are subtle but definitive. Regardless of the advancement, there is a long way to go, as customized techniques are required to be fine-tuned with precision to attain more insights into tumor reversion. Tumor regression models using available genome-editing methods, followed by in vitro and in vivo proteomics profiling techniques show early evidence. This review summarizes tumor reversion developments, present issues, and unaddressed challenges that remained in the uncharted territory to modulate cellular machinery for tumor reversion towards therapeutic purposes successfully. Ongoing research reaffirms the potential promises of understanding the mechanism of tumor reversion and required refinement that is warranted in vitro and in vivo models of tumor reversion, and the potential translation of these into cancer therapy. Furthermore, therapeutic compounds were reported to induce phenotypic changes in cancer cells into normal cells, which will contribute in understanding the mechanism of tumor reversion. Altogether, the efforts collectively suggest that tumor reversion will likely reveal a new wave of therapeutic discoveries that will significantly impact clinical practice in cancer therapy.

Possible Mechanisms of Liver Injury Induced by Cadmium Sulfide Nanoparticles in Rat.

Cadmium is primarily utilized in the construction of particles known as quantum dots. Hepatotoxicity caused by microparticles of cadmium is very well known; however, toxicity of nanoparticles of cadmium is not well understood. The present study describes the toxicity of cadmium sulfide nanoparticles (CdSNPs) in the liver of rat. Adult Wistar rats were administered CdSNPs (10 mg/kg) on alternate days for 45 days. Serum enzymes (ALT, AST, ALP), biomarkers of lipid peroxidation (MDA, H2O2, and NO), and metallothionein concentration were determined. Histopathological and TEM observations were also made to record morphological changes. CdSNPs (10 mg/kg) induced significant changes in the structure and function of liver. Values of serum enzymes and reactive species increased significantly in rats treated with CdSNPs in comparison to CdS-treated rats. Histopathological observations showed extensive parenchymal degeneration. Ultrastructural studies exhibited proliferation of endoplasmic reticulum, microsomes, and lysosomes. It is concluded that NP-membrane interaction leads to the generation of reactive species that alter membrane integrity and induce oxidative stress. These events may activate cell death pathways in hepatocytes.

Distinctive fingerprints of genotoxicity induced by As, Cr, Cd, and Ni in a freshwater fish.

Genotoxicity of three toxic elements (chromium, cadmium, nickel) and a metalloid (arsenic) has been studied in a freshwater fish, Channa punctatus using micronuclei (MN) test, comet assay, and erythrocyte nuclear alterations (ENAs) as fingerprints of genotoxicity. These tests yielded different results suggesting involvement of different mechanisms for their genotoxicity. While highest frequency of blebbed nuclei was observed in chromium-treated fish (6.5 ± 0.76), lowest was observed in cadmium-treated fish (4.0 ± 1.0). Maximum number of notched nuclei was recorded in arsenic-treated fish (5.5 ± 1.15) whereas highest numbers of lobed nuclei were found in cadmium-treated fish (4.5 ± 0.13). These differences might be attributed to selective bioaccumulation and chemodynamics of each element. Other parameters used to determine genotoxicity viz.: lipid peroxidation and DNA damage also suggested different mechanisms of their genotoxicity. It is suggested that an integrative approach, using a battery of tests for determining genotoxicity, should be made while making environmental health risk assessment and ecotoxicological studies of these toxic elements.

Renal toxicity of nanoparticles of cadmium sulphide in rat.

During present investigations, renal toxicity of CdSNPs (cadmium sulphide nanoparticles) (ranged 5-9 nm) was estimated in rat employing specific parameters. Treatment on each alternate day for 45 days with CdSNPs (10 mg/kg b.w.) yielded significant increase in Cd-MT (cadmium metallothionein), lipid peroxidation and H2O2 generation in the kidney of rat in comparison to bulk cadmium. Concentration of creatinine also increased in urine of CdSNPs treated rats. Histopathological observations revealed extensive damage in proximal tubules. Ultrastructural studies showed mitochondrial, nuclear and ER (endoplasmic reticulum) changes. Finally, renal toxicity of CdSNPs was confirmed by loss of alkaline phosphatase from the brush border of proximal convoluted tubules. These observations conclude that CdSNPs manifest greater toxicity in kidney than cadmium sulphide bulk particles. These effects have been attributed to their specific physicochemical properties, greater potential to generate ROS and induction of oxidative stress and impairment of renal structure and function. Present observations suggest that commercial/industrial use of CdSNPs may pose serious renal health problems in man.

Zinc oxide nanoparticles inhibit dimethylnitrosamine induced liver injury in rat.

Dimethylnitrosamine (DMN) is a potent hepatotoxic, carcinogenic and mutagenic compound. It induces massive liver cell necrosis and death in experimental animals. Several drugs have been tested in the past for their protective behavior against DMN toxicity. However, it is for the first time that therapeutic intervention of ZnONPs (zinc oxide nanoparticles) has been studied against its toxicity. Present results show that a post treatment of ZnONPs (50 mg/kg) to DMN (2 µl/100 g body weight) treated rats reduces lipid peroxidation, oxidative stress and fibrosis in the liver. It diminishes serum ALT (alanine transaminases), AST (aspartate transaminases) and LDH (lactate dehydrogenase) showing improvement in liver function. Reduced values of proinflammatory cytokines viz. TNF-α and IL-12 also support its protective effects. Histopathological observations also indicate improvement in liver cell morphology. It is postulated that ZnONPs offer protection through selective toxicity to proliferating tissue including adenomatous islands formed in the liver. Zinc metallothionein (Zn-MT) induced by ZnONPs may also contribute in the amelioration of DMN induced toxic effects. Diminution of oxidative stress by ZnONPs remains to be the key mechanism involved in its protective effects. However, toxicity of ZnONPs in the liver needs to be monitored simultaneously.

Assessment of genotoxicity amongst smokers, alcoholics, and tobacco chewers of North India using micronucleus assay and urinary 8-hydroxyl-2'-deoxyguanosine, as biomarkers.

The main objective of the present study was to screen the genotoxicity caused by individual and combined habits of smoking, tobacco chewing, and alcohol consumption in human population of North India. Study recruited 67 male subjects aged 25 to 65 years. Buccal mucosal cells were subjected to micronucleus (MN) assay, and 8-hydroxyl-2-deoxyguanosine (8-OHdG) was estimated in their urine samples. Number and shape of the MN cells varied in the buccal epithelium of different groups. Maximum number of MN (0.47%) were found in tobacco chewers followed by smokers (0.45%) and alcoholics (0.44%) (P < 0.05). These results reciprocated the concentration of urinary 8-OHdG. Maximum value for 8-OHdG was also recorded in tobacco chewers (21.07 ± 5.51 mg/mg creatinine) followed by smokers (20.25 ± 3.96 mg/mg creatinine) and alcoholics (19.06 ± 3.41 mg/mg creatinine) (P < 0.05). Combined effects of these agents were found to be statistically different from individual effects. Carcinogenic compounds present in cigarette smoke, nitrosamines found in solid tobacco, and acetaldehyde, a metabolic product of alcohol, induce oxidative stress that manifests into genotoxicity. In conclusion, demographical differences occur in the genotoxicity caused by these three habits. MN assay and urinary 8-OHdG are simple, noninvasive, and reliable biomarkers of genotoxicity.

Modulation of phase-II enzyme activities in benzene treated ovariectomized rats.

The aim of the study was to determine the influence of ovariectomy on phase II enzymes viz. glutathione-S-transferase (GST), glutathione peroxidase (GPX) and catalase (CAT) in liver and kidney of female rats treated with benzene. The results showed the significant decrease of the GST and GPX activity in benzene treated rats after ovariectomy. However progesterone supplementation stimulated the activity of GST and GPX in liver and kidney of benzene treated non ovariectomized and ovariectomized rats. Progesterone supplementation to benzene treated ovariectomized rats helps to gain in CAT activity. Our results on DNA damage using single cell gel electrophoresis also confirmed our findings on antioxidant enzymes. The results showed that lack of protective progesterone against benzene toxicity is reflected in alterations in antioxidant enzyme activities. However progesterone therapy to benzene treated ovariectomized rats results in activating the antioxidant defence system. Since female workers are engaged in industrial sector, these results are important from occupational health point of view. Benzene exposure affects their reproductive health. Nevertheless, it could be modulated by suitable hormonal therapy.

Endocrinal toxicity of industrial solvents--a mini review.

Endocrine system can be affected by various organic compounds. The review describes the effects of major industrial solvents on adrenal, thyroid and parathyroid glands in man and experimental animals. Further, their toxicity in pancreas, pituitary, testis and ovary has also been discussed. An attempt has been made to offer a historical and general information on solvent toxicity in endocrine glands. Possible mechanisms, in brief, have also been discussed. Endocrine toxicity caused by industrial solvents deserves more attention than hitherto paid. An understanding of hormonal disorders caused by industrial solvents will be important from occupational health point of view.

Modulation of CYP4502E1 and oxidative stress by testosterone in liver and kidney of benzene treated rats.

Bilateral castration increased lipid peroxidation and consequently reduced glutathione in both liver and kidney. Testosterone administration reduced lipid peroxidation in the liver of castrated and benzene treated rats, however, reduced glutathione status could not be restored. Benzene depleted CYP4502E1 in castrated rats, however, the enzyme was restored in liver and kidney both after testosterone treatment. The results suggest that testosterone affects the metabolism and disposition of benzene by influencing CYP4502E1. Other hormonal and cellular/molecular factors may also alter the actions of testosterone. Testosterone dependent mechanism of toxicity of benzene in the liver and kidney has been discussed.

Effects of progesterone on benzene toxicity in rats.

Benzene is a frequently used industrial solvent. Its toxic manifestations could be modified by sex hormones, but mechanisms of their action are poorly understood. We have examined the influence of progesterone on lipid peroxidation (malondialdehyde), reduced glutathione (GSH), and cytochrome P450 2E1 (CYP2E1) in the liver and kidneys of female rats. Progesterone applied to benzene-treated rats inhibited the formation of reactive oxygen species (ROS), but in ovariectomised benzene-treated rats it significantly increased GSH in the liver. No improvement in CYP2E1 activity was observed in progesterone treated rats. Our results evidence that progesterone changes benzene toxicity (generation of ROS, oxidative stress). However, the probable antioxidative effect of progesterone needs to be confirmed by further studies.

Circadian variation in lipid peroxidation induced by benzene in rats.

Time-dependent effect of benzene, a potent carcinogenic industrial solvent, on lipid peroxidaiton and associated mechanisms has been studied in liver and kidney of rats. Significant differences were observed in the values of urinary phenol, microsomal malondialdehyde, reduced glutathione (GSH) and cytochrome P4502E1 in rats treated with benzene in morning and evening hours. Higher were the values for urinary phenol and hepatic microsomal malondialdehyde in rats administered benzene in evening hours. Contrarily, higher were the values for GSH and cytochrome P4502E1 in rats treated with benzene in morning hours. Increased microsomal lipid peroxidation has been attributed to low GSH status, whereas increased phenol concentration could be related to low activity of cytochrome P4502E1 in the liver of rats in evening hours. It is concluded that circadian rhythmicity in hepatic drug metabolizing enzyme system and GSH contributes in toxicity of benzene. The results are important from occupational health point of view.

Biochemical toxicity of benzene.

Human exposure to benzene in work environment is a global occupational health problem. After inhalation or absorption, benzene targets organs viz. liver, kidney, lung, heart and brain etc. It is metabolized mainly in the liver by cytochrome P450 multifunctional oxygenase system. Benzene causes haematotoxicity through its phenolic metabolites that act in concert to produce DNA strand breaks, chromosomal damage, sister chromatid exchange, inhibition of topoisomerase II and damage to mitotic spindle. The carcinogenic and myelotoxic effects of benzene are associated with free radical formation either as benzene metabolites or lipid peroxidation products. Benzene oxide and phenol have been considered as proheptons. Liver microsomes play an important role in biotransformation of benzene whereas in kidney, it produces degenerative intracellular changes. Cohort studies made in different countries suggest that benzene induces multiple myeloma in petrochemical workers. Though extensive studies have been performed on its toxicity, endocrinal disruption caused by benzene remains poorly known. Transgenic cytochrome P450 IIE1 mice may help in understanding further toxic manifestations of benzene.

Sex differences in oxidative stress induced by benzene in rats.

Role of sex differences on oxidative stress induced by benzene has been studied in liver, kidney and lungs of rat. It was observed that benzene administration enhanced lipid peroxidation in liver, kidney and lungs of rat, nevertheless, significant variations were recorded in male and female rats. Decrease of GSH and CYTP(450)2E1 was higher in female rats than male rats except lungs. The results suggest that oxidative stress induced by benzene is higher in female rats.

Gender differences in the metabolism of benzene, toluene and trichloroethylene in rat with special reference to certain biochemical parameters.

Metabolites viz. phenol, hippuric acid and total trichloro compounds of benzene, toluene and trichloroethylene respectively were estimated in the urine samples of male and female rats after exposure for a period of 30 days. The results exhibited higher metabolism in female rats than the male rats. Their metabolism might be regulated by cytochrome P450 isozymes in a gender specific manner. However, sex differences in the activity of glutathione-S-transferases of the liver have also been found to determine their toxicity. Results have been discussed with quantitative profiles of other enzymes established in the liver of male and female rats.