A stable dark variant separated from photobacterium phosphoreum (A2) was fixed in agar-gel membrane and immobilized onto an exposed end of a fiber-optic linked with bioluminometer. The variant could emit a luminescent...A stable dark variant separated from photobacterium phosphoreum (A2) was fixed in agar-gel membrane and immobilized onto an exposed end of a fiber-optic linked with bioluminometer. The variant could emit a luminescent signal in the presence of genotoxic agents, such as Mitomycin C (MC). The performance of this whole-cell optical fiber sensor system was examined as a function of several parameters, including gel probe thickness, bacterial cell density, and diameter of the fiber-optic core and working temperature. An optimal response to a model genotoxicant, Mitomycin C, was achieved with agar-bacterial gel membrane: the thickness of gel membrane was about 5 mm; the cell density of bacteria in gel membrane was about 2.0×107 /ml; the diameter of fiber-optic core was 5.0 mm; the working temperature was 25 ℃. Under these optimized conditions, the response time was less than 10 h to Mitomycin C, with a lower detection threshold of 0.1 mg/L.展开更多
Metallothioneins （MTs） were discovered in 1957 by Margoshes and Vallee and identified as low-molecular weight and sulphydryl rich proteins. It is not surprising that most mammalian tissues contain age related basal ...Metallothioneins （MTs） were discovered in 1957 by Margoshes and Vallee and identified as low-molecular weight and sulphydryl rich proteins. It is not surprising that most mammalian tissues contain age related basal levels of MTs since they are involved in metalloregulatory processes that include cell growth and multiplication. In an effort to understand the biology of this intriguing tumor, various biomarkers such as oncogenes, p53 tumor suppressor gene, war 1 protein, proliferating cell nuclear antigen, telomerase, microsatellite markers and cytogenetic changes have been examined. One biomarker which has recently shown to be expressed in various human tumors but still less reported in carcinoma is MT. Immunohistochemical detection of MT proteins in cold acetone-fixed paraffin embedded liver sections was performed by the streptavidin-avidin-biotin immunoperoxidase complex method.展开更多
Objective: To assess and compare the cytotoxic, genotoxic, apoptotic and reactive oxygen species(ROS) generating effects of naringenin(NG) and its new derived compound naringenin-oxime(NG-Ox) on MCF-7, HT-29, PC-12 ca...Objective: To assess and compare the cytotoxic, genotoxic, apoptotic and reactive oxygen species(ROS) generating effects of naringenin(NG) and its new derived compound naringenin-oxime(NG-Ox) on MCF-7, HT-29, PC-12 cancer and L-929 normal cell lines.Methods: The cells were incubated with different doses of NG-Ox and NG(50–1 000 mmol/L) for 24 h. The cell viability was assessed based on ATP cell viability assay.Intracellular accumulation of ROS was determined using the fluorescent probes 2070-dichlorodihydrofluorescin diacetate. Genotoxic effects were evaluated by alkaline single cell gel electrophoresis assay(comet assay) and, the apoptotic effect was evaluated by acridine orange staining at below the IC50 levels.Results: Both NG-Ox and NG exhibited cytotoxic, genotoxic and apoptotic effects and resulted in increased ROS values in a dose-dependent manner. The effects were more pronounced on cancer cell lines. The cytotoxic, genotoxic and apoptotic effects of NG-Ox were higher than that of NG in all cell lines. Significant correlations were observed between cell viability, DNA damage, apoptosis and ROS, in all cell lines exposed to either NG-Ox or NG.Conclusions: This study showed that both NG-Ox and NG possess cytotoxic, genotoxic and apoptotic activities through the production of ROS on cells, NG-Ox being the more effective one. Therefore, derived compound of NG might be used as antiproliferative agents for the treatment of cancer.展开更多
Covalent organic nanospheres(CONs)were explored as a fiber coating for solid-phase microextraction of genotoxic impurities(GTIs)from active ingredients(AIs).CONs were synthesized by an easy solutionphase procedure at...Covalent organic nanospheres(CONs)were explored as a fiber coating for solid-phase microextraction of genotoxic impurities(GTIs)from active ingredients(AIs).CONs were synthesized by an easy solutionphase procedure at 25℃.The obtained nanospheres exhibited a high specific surface area,good thermostability,high acid and alkali resistance,and favorable crystallinity and porosity.Two types of GTIs,alkyl halides(1-iodooctane,1-chlorobenzene,1-bromododecane,1,2-dichlorobenzene,1-bromooctane,1-chlorohexane,and 1,8-dibromooctane)and sulfonate esters(methyl p-toluenesulfonate and ethyl ptoluenesulfonate),were chosen as target molecules for assessing the performance of the coating.The prepared coating achieved high enhancement factors(5097-9799)for the selected GTIs.The strong affinity between CONs and GTIs was tentatively attributed to π-π and hydrophobicity interactions,large surface area of the CONs,and size-matching of the materials.Combined with gas chromatography-mass spectrometry(GC-MS),the established analytical method detected the GTIs in capecitabine and imatinib mesylate samples over a wide linear range(0.2-200 ng/g)with a low detection limit(0.04-2.0 ng/g),satisfactory recovery(80.03%-109.5%),and high repeatability(6.20%-14.8%)and reproducibility(6.20%-14.1%).Therefore,the CON-coated fibers are promising alternatives for the sensitive detection of GTIs in AI samples.展开更多
Human pluripotent stem cells(h PSCs) represent heterogeneous populations, including induced pluripotent stem cells(i PSCs), endogenous plastic somatic cells, and embryonic stem cells(ESCs). Human ESCs are derived from...Human pluripotent stem cells(h PSCs) represent heterogeneous populations, including induced pluripotent stem cells(i PSCs), endogenous plastic somatic cells, and embryonic stem cells(ESCs). Human ESCs are derived from the inner cell mass of the blastocyst, and they are characterized by the abilities to self-renew indefinitely, and to give rise to all cell types of embryonic lineage(pluripotency) under the guidance of the appropriate chemical, mechanical and environmental cues. The combination of these critical features is unique to h ESCs, and set them apart from other human cells. The expectations are high to utilize h ESCs for treating injuries and degenerative diseases; for modeling of complex illnesses and development; for screening and testing of pharmacological products; and for examining toxicity, mutagenicity, teratogenicity, and potential carcinogenic effects of a variety of environmental factors, including ionizing radiation(IR). Exposures to genotoxic stresses, such as background IR, are unavoidable; moreover, IR is widely used in diagnostic and therapeutic procedures in medicine on a routine basis. One of the key outcomes of cell exposures to IR is the change in gene expression, which may underlie the ultimate h ESCs fate after such a stress. However, gaps in our knowledge about basic biology of h ESCs impose a serious limitation to fully realize the potential of h ESCs in practice. The purpose of this review is to examine the available evidence of alterations in gene expression in human pluripotent stem cells after genotoxic stress, and to discuss strategies for future research in this important area.展开更多
In the present study, a disposable electrochemical DNA nano-biosensor is proposed for the rapid detection of genotoxic compounds and bio-analysis of water pollution. The DNA nano-biosensor is prepared by immobilizing ...In the present study, a disposable electrochemical DNA nano-biosensor is proposed for the rapid detection of genotoxic compounds and bio-analysis of water pollution. The DNA nano-biosensor is prepared by immobilizing DNA on Au nanoparticles and a self-assembled monolayer of cysteamine modified Au electrode. The assembly processes of cysteamine, Au nanoparticles and DNA were characterized by cyclic voltammetry （CV）. The Au nanoparticles enhanced DNA immobilization resulting in an increased guanine signal. The interaction of the analyte with the immobilized DNA was measured through the variation of the electrochemical signal of guanine by square wave voltammetry （SWV）. The biosensor was able to detect the known genotoxic compounds： 2-anthramine, acridine orange and 2- naphthylamine with detection limits of 2, 3 and 50 nmol/L, respectively. The biosensor was also used to test actual water samples to evaluate the contamination level. Additionally, the comparison of results from the classical genotoxiciw bioassay has confirmed the applicability of the method for real samoles.展开更多
Highly sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed for the simultaneous determination of five potential genotoxic impurities in ranolazine active pharmaceutical ingred...Highly sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed for the simultaneous determination of five potential genotoxic impurities in ranolazine active pharmaceutical ingredient. Chromatographic separation achieved using Poroshell C18 PFP 150 × 3.0 mm 2.7 μ column and 0.1% formic acid in water as Mobile phase A and 0.1% formic acid in methanol as mobile phase B using gradient elution and a flow rate of 0.4 ml/min with a run time of 18 minutes. Mass spectrometric conditions were optimized using electrospray ionization in positive mode. Method shows excellent linearity from 0.05 - 5.0 ppm of the ranolazine test concentration for all the five impurities. The correlation coefficient was observed greater than 0.99. Satisfactory recoveries were observed for all the five impurities within the range of 102.9% - 112.3%. Method has been validated as per ICH recommended guidelines with a LOQ of 0.15 ppm achieved. The developed method was able to quantify all the five impurities at a concentration level of 1 ng/ml (0.5 ppm with respect to 2 mg/ml ranolazine).展开更多
Nitrosamine impurities are potentially genotoxic which are considered under cohort of concern as per ICH M7 guidelines and need to be controlled at trace levels during quantification in drug substances and drug produc...Nitrosamine impurities are potentially genotoxic which are considered under cohort of concern as per ICH M7 guidelines and need to be controlled at trace levels during quantification in drug substances and drug products for safe human consumption. Recent regulatory requirements also suggest the need to have highly sensitive analytical methods for the accurate quantification of Nitrosamine impurities. In this paper we have presented simple, rapid and ultra-sensitive LC-MS/MS method for six potential genotoxic nitrosamine impurities: N-Nitroso dimethyl amine (NDMA), N-Nitroso diethyl amine (NDEA), N-Nitroso Ethyl Iso propylamine (NEIPA), N-Nitroso-N-methyl-4-aminobutyric acid (NMBA) N-Nitroso diisopropylamino (NDIPA) and N-Nitroso dibutyl amine (NDBA) with a LOQ of 0.004 ppm. Chromatographic separation is achieved using Zorbax SB C18 150 × 3.0 mm, 3.5 μ column with 0.1% formic acid in water as mobile phase A and 0.1% formic acid in methanol as mobile phase B at a flow rate of 0.3 ml/min using gradient mode of elution at a total run time of 18 minutes. Six nitrosamine impurities are successfully ionized and quantified in positive mode of atmospheric pressure chemical ionization (APCI) using multiple reaction monitoring (MRM). Method validation is performed as per ICH guidelines evaluating the limit of quantification and detection and found to give good S/N ratios with good linearity range of 0.002 - 2 ppm with regression coefficient >0.99 for all the six nitrosamine impurities. Method recoveries are established using three-step sample preparation protocol and are found to be satisfactory within 80% - 120%. The method can be used routinely applied for the detection of Nitrosamines in Telmisartan at a concentration of 1.5 ng/ml (0.03 ppm with respect to telmisartan concentration of 50 mg/ml).展开更多
Defining impurity profile is key element to ensure safe, efficacious and quality human drugs. Impurity profiling changed/transformed drastically over the years. Guidelines, specifications and requirements are evolving...Defining impurity profile is key element to ensure safe, efficacious and quality human drugs. Impurity profiling changed/transformed drastically over the years. Guidelines, specifications and requirements are evolving. Initially impurity profiling was based on simple methods later by degradation studies, then to understand drug strength and efficacy chiral impurities and stereo isomers were included followed by residual solvents, polymorphic forms, genotoxic impurity studies. Currently, elemental impurities are the latest addition. As per the GDUFA II guidelines to improve review efficiency and reduce review cycles, data requirements have changed. Based on recent guidance and review points, Impurity profiling has significant importance in ANDA filing and to ensure approval within 10 months (first cycle approval) which is an exiling aspect for industries to enter into the generic market quickly. Hence, Impurity profile is a key aspect scientifically, regulatory wise and commercially also. This is a review article on impurity profiling of Solid oral drug substances and products as per GDUFA II requirements the reference documents for the review are ICH guidance, relevant FDA GDUFA guidance and common industry practices.展开更多
In this study, two common pollutants (benzo[a]pyrene and methamidophos) in marine environment were tested by comet assay for their inducement of in vivo genotoxic effect to the blood cells of black porgy ( Acanthopagr...In this study, two common pollutants (benzo[a]pyrene and methamidophos) in marine environment were tested by comet assay for their inducement of in vivo genotoxic effect to the blood cells of black porgy ( Acanthopagrus schlegeli). The fish was exposed to 2 μg/L of benzo[a]pyrene (BaP) and methamidophos, and their mixture. The assay was performed on whole blood at 2 h, 5 h, 24 h and 96 h exposure intervals. A signifi- cant increase in DNA damage was observed in each treatment with the pollutants. Additive effect of BaP and methamidophos was also found in the experiment. However, the decrease ratios of DNA damage for 5 h and 96 h exposure interals compared with 2 h and 24 h exposure ones, respectively, were noticed. This phenomenon may be explained by the function of repairing process via enzyme cytochrome P450 in the animal. Evidence of the genotoxicity of organophosphorus pesticides (OPs) and polynuclear aromatic hydrocarbons (PAHs) on marine fish are discussed in this paper.展开更多
A series of bioassays such as sister chromatid exchange frequencies ( SCE.), chromosomal aberration ( CA ), micronuclel rate (MN) and cell-cycle delay have been used to detecting the genotoxic effect of cigarette smok...A series of bioassays such as sister chromatid exchange frequencies ( SCE.), chromosomal aberration ( CA ), micronuclel rate (MN) and cell-cycle delay have been used to detecting the genotoxic effect of cigarette smoke condensate (CSC) on human diploid cell 2BS strain. The results suggested that a higher SCE, ( 17. 0/ cell) was observed In 2BS cells treated with CSC at 100 μg/ml, as compared with 6. 9/cell of the background (P<0. 001). CA rate was significantly increased from 4% to 36% In cells treated with 10 μg/ml CSC (P< 0.001). MN rate varied from 9 -26‰ In cells treated with CSC compared to that of control (6‰). Meanwhile, the cell-cycle of cells was markedly delayed by CSC. The survival rate of 2BS cells declined to 59. 6% for treatment with CSC at 200 μg/ ml. There was a dose-effect response In SCE., CA, MN rate. We proposed that active oxygen might responsible for genotoxiclty of CSC on cells.展开更多
Medicinal use of spices/herbs has been gradually increased in the developed countries, Zingiber officinale （Ginger） is known to possess potent antioxidant and anti inflammatory properties. Therefore, the aim of this...Medicinal use of spices/herbs has been gradually increased in the developed countries, Zingiber officinale （Ginger） is known to possess potent antioxidant and anti inflammatory properties. Therefore, the aim of this study is to determine the possible anti-mutagenic effect of ginger against the genotoxic effect of anti-cancer drug Taxol 0.6 mg/kg. This study is conducted by using two types of cytogenetic studies in bone marrow cell of mal albino mice Mus musculus （average weight 25-30 g）. The animals were randomly distributed into six groups, each of 14 mi[ce, （GI） was given the solvent, （G2） treatment of the medical dose of Taxol drug, （G3） treatment of ginger, （G4） a pre-treatment of ginger prior to treatment of drug, （G5） a simultaneous treatment of ginger and treatment of drug, （G6） a post-treatment of ginger after treatment drug. The study results show that significant increase in total chromosomal aberrations and significant increase in the number of micronuclei were observed after treatment drug. The significant structural aberrations were in the form of end-to-end associations. The numerical chromosomal aberrations were endomitosis and polyploid. The results showed that the frequencies of chromosomal aberrations and micronuclei in ginger treated group were not significantly different from control. Simultaneous treatment of ginger was found to be effective in reducing the genotoxic effects induced by drug Taxol especially in the total number of the chromosomal aberrations and the number of micronuclei.展开更多
A simple, rapid, and highly sensitive LC-MS/MS method has been developed for the simultaneous and trace level quantification of underivatized boronic acids in lumacaftor active pharmaceutical ingredient. Chromatograph...A simple, rapid, and highly sensitive LC-MS/MS method has been developed for the simultaneous and trace level quantification of underivatized boronic acids in lumacaftor active pharmaceutical ingredient. Chromatographic separation of boronic acids and lumacaftor achieved using Agilent Poroshell HPH C18 150<span> </span><span>×</span><span> </span><span>4.6</span><span> </span><span>mm 2.7</span><span> </span><span>μ column with 0.1% ammonia in water as mobile phase A and 100%</span><span> </span><span>acetonitrile as mobile phase B at a flow rate of 0.25</span><span> </span><span>ml/min. Gradient elution</span><span> was</span><span> used with a total method run time of 14</span><span> </span><span>minutes. Boronic acids were successfully ionized and quantified without derivatization using electrospray ionization in negative mode using tandem quadrupole mass spectrometry in multiple reactions monitoring mode. Method validation </span><span>was </span><span>performed as per ICH guidelines with good linearity over the concentration range of 0.05 ppm to 5 ppm of Lumacaftor test concentration for both the boronic acids with a correlation coefficient of >0.99.</span><span> </span><span>Recoveries were found good at different concentration levels and within the range of 80</span><span>% </span><span>-</span><span> </span><span>120%.</span><span> </span><span>The developed method can be successfully used for the routine quantification of boronic acids at a concentration level of 20</span><span> </span><span>ng/ml (1</span><span> </span><span>ppm with respect to 20</span><span> </span><span>mg/ml lumacaftor).</span>展开更多
The genotoxic activities of effluents from drainage water treatment plants were examined by using the novel <em>umu</em> tester strain NM8001, which lacks <em>MutMst</em> genes. To enhance the ...The genotoxic activities of effluents from drainage water treatment plants were examined by using the novel <em>umu</em> tester strain NM8001, which lacks <em>MutMst</em> genes. To enhance the sensitivity of the LacZ assay, a BugBuster mix protein extraction reagent and TokyoGreen-<em>β</em> Gal for a fluorescence-galactosidase substrate were applied. Of the 24 sampling locations present in Kanagawa prefecture, Japan, water extracts from nine sampling points showed apparent genotoxic activities without metabolic activation. In contrast, water extracts from the upper sites of these water treatment plants did not show any significant genotoxic activities. The selected samples with genotoxic activity did not show significant mutagenicity toward Ames strains TA98 and TA100. Genotoxicity was also well correlated with the activity of a classical <em>umu </em>strain of TA1535/pSK1002;these findings indicate that the genotoxicity induced by oxidative damage was not a significant component of the genotoxicity.展开更多
The no-observed-effect level (NOEL) in a study of carcinogenicity for compounds that are both genotoxic and carcinogenic represents the limit of detection in that bioassay, rather than an estimate of a possible thresh...The no-observed-effect level (NOEL) in a study of carcinogenicity for compounds that are both genotoxic and carcinogenic represents the limit of detection in that bioassay, rather than an estimate of a possible threshold. Therefore, for those genotoxic and carcinogenic contaminants (e.g. acrylamides, PAHs, etc.) in foods it is not possible to develop health-based guidance values (e.g. ADI or PTWI) using the traditional NOEL and safety/uncertainty factors.展开更多
4-hydrazinobenzoic acid has been highlighted as one of the potential genotoxic impurities (PGIs). A sensitive HPLC method was developed and validated for the determination of 4-hydrazino benzoic acid in Deferasirox Ta...4-hydrazinobenzoic acid has been highlighted as one of the potential genotoxic impurities (PGIs). A sensitive HPLC method was developed and validated for the determination of 4-hydrazino benzoic acid in Deferasirox Tablet. HPLC method on Zorbax SB C18 column (250 × 4.6 mm i.d.), 5 μm, with UV Detector was used. The proposed method was cost effective, specific, linear, accurate, rugged and precise. The calibration curves showed good linearity over the concentration range of 0.5 μg/ml to 1.5 μg/ml with respect to the sample. The correlation coefficient was 0.999. Excellent recoveries of 102% were obtained at the level 0.5 μg/ml.展开更多
With the ever increasing complexity of active pharmaceutical ingredient (API) preparations, more potential genotoxic impurities (PGI’s) are being observed. It is thus necessary to determine if these PGI’s are presen...With the ever increasing complexity of active pharmaceutical ingredient (API) preparations, more potential genotoxic impurities (PGI’s) are being observed. It is thus necessary to determine if these PGI’s are present in the final API’s, and if they are present, to ensure the levels are acceptable for any clinical uses. For PGI’s that have authentic standards available, quantitation can be accomplished in a straightforward manner. However, for PGI’s that are expected to form through rearrangements or side reactions, authentic standards may not be readily available, significantly complicating the analysis. In this study we describe a surrogate standard approach for quantifying PGI’s that allows for relative response factor calculations of PGI species utilizing both gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS).展开更多
Background: In oral and maxillofacial surgery, synthetic bone grafts are most widely used as bone substitutes, due to the limited sources of autologous bone. The aim of this study was to examine the influence of three...Background: In oral and maxillofacial surgery, synthetic bone grafts are most widely used as bone substitutes, due to the limited sources of autologous bone. The aim of this study was to examine the influence of three different synthetic bone grafts (Cerasorb, Fortoss and Perioglass) on sisters chromatide exchanges (SCEs) in peripheral lymphocytes. Materials and Methods: Peripheral blood samples taken from 68 patients (45 females and 23 males), who underwent oral surgery procedures, such as apical resection, cyst enucleation or periodontal curretage, were obtained for SCE a day before and two months after the surgeries. A control group included 30 patients, while the study group was made of the patients who underwent bone grafting with Cerasorb? (11 patients), Fortoss? VITAL (10 patients) or Perioglass? (17 patients). Results: Comparing with the results of the study group before and after the treatment, it was concluded that the results were statistically significant (p = 0.001). In the Perioglass? subgroup, a greater statistical significance (p = 0.003) was noted, than that in either the Cerasorb? (p = 0.620) or Fortoss? (p = 0.210) subgroups, in which there was no statistical significance. Conclusions: Although further investigations may be necessary, our results suggest that the synthetic bone grafts might have an influence on SCE in peripheral lymphocytes.展开更多
Population growth and the increase in the consumption of different pharmaceuticals combined with the insufficiency in the removal of these compounds by conventional treatments have contributed to the increase in the d...Population growth and the increase in the consumption of different pharmaceuticals combined with the insufficiency in the removal of these compounds by conventional treatments have contributed to the increase in the detection of these contaminants in aquatic matrices.Aiming to contribute in solving this problem,this promoted the degradation of a mixture of the drugs lamivudine and zidovudine in different matrices(aqueous solution and synthetic effluent)using the heterogeneous photo-Fenton process applying pyrite as a catalyst and artificial solar radiation.At the end of the treatment,degradations greater than 99%were found for zidovudine in both matrices studied,while for lamivudine,97%and 94%degradations were obtained for aqueous solution and synthetic effluent,in that order.In the investigation of toxic effects using Biomphalaria glabrata molluscs,embryotoxicity tests showed embryonic lethality in 100%of individuals for all samples.Acute toxicity tests on adult molluscs resulted in mortality rates of 100%(aqueous solution after treatment)and 50%(synthetic effluent after treatment).Thus,to investigate cellular changes,genotoxicity analyses were carried out,and different degrees of DNA damage were observed,however,the highest level of damage to this organism was not observed.Therefore,B.glabrata demonstrated to be sensitive to toxic effects at the concentrations present in the matrices studied,providing evidence to predict the ecotoxicological potential of samples when released into aquatic ecosystems.展开更多
Green synthesis of silver nanoparticles(NPs)by green route approaches has advantages over conventional methods.In green synthesis,we use eco-friendly plant extracts contain secondary metabolites and bioactive componen...Green synthesis of silver nanoparticles(NPs)by green route approaches has advantages over conventional methods.In green synthesis,we use eco-friendly plant extracts contain secondary metabolites and bioactive components,proteins that act as both reducing and capping agents,form stable and shape-controlled green silver nanoparticles.The current study deals with the synthesis of silver nanoparticles using the aqueous latex extract of Allamanda cathartica.The green silver nanoparticles are characterized by using different spectroscopic methods like ultra violet-visible spectroscopy(UV-Vis),Fourier transform-infrared spectroscopy(FTIR),transmission electron microscope(TEM),scanning electron microscope(SEM)and X-ray diffraction(XRD).Results indicated that the crystalline natured particles were spherical shaped with an average of 35 nm in size,and that the stability of silver nanoparticles was due to its high negative zeta potential of–27.6 mV.The current study also revealed that green silver nanoparticles had very good genotoxic and cytotoxic activity in peripheral blood mononuclear cells(PBMCs).Leukemia leads to the development of high numbers of white blood cells,which is one of the major types of cancers that affect children.Many of the chemicals used for the treatment produce remarkable side effects.To overcome this problem,we made an attempt to see the efficacy of latex green silver nanoparticle on peripheral blood mononuclear cells and deoxyribonucleic acid fragmentation,which leads to the development of future therapeutic drugs.展开更多
文摘A stable dark variant separated from photobacterium phosphoreum (A2) was fixed in agar-gel membrane and immobilized onto an exposed end of a fiber-optic linked with bioluminometer. The variant could emit a luminescent signal in the presence of genotoxic agents, such as Mitomycin C (MC). The performance of this whole-cell optical fiber sensor system was examined as a function of several parameters, including gel probe thickness, bacterial cell density, and diameter of the fiber-optic core and working temperature. An optimal response to a model genotoxicant, Mitomycin C, was achieved with agar-bacterial gel membrane: the thickness of gel membrane was about 5 mm; the cell density of bacteria in gel membrane was about 2.0×107 /ml; the diameter of fiber-optic core was 5.0 mm; the working temperature was 25 ℃. Under these optimized conditions, the response time was less than 10 h to Mitomycin C, with a lower detection threshold of 0.1 mg/L.
文摘Metallothioneins （MTs） were discovered in 1957 by Margoshes and Vallee and identified as low-molecular weight and sulphydryl rich proteins. It is not surprising that most mammalian tissues contain age related basal levels of MTs since they are involved in metalloregulatory processes that include cell growth and multiplication. In an effort to understand the biology of this intriguing tumor, various biomarkers such as oncogenes, p53 tumor suppressor gene, war 1 protein, proliferating cell nuclear antigen, telomerase, microsatellite markers and cytogenetic changes have been examined. One biomarker which has recently shown to be expressed in various human tumors but still less reported in carcinoma is MT. Immunohistochemical detection of MT proteins in cold acetone-fixed paraffin embedded liver sections was performed by the streptavidin-avidin-biotin immunoperoxidase complex method.
文摘Objective: To assess and compare the cytotoxic, genotoxic, apoptotic and reactive oxygen species(ROS) generating effects of naringenin(NG) and its new derived compound naringenin-oxime(NG-Ox) on MCF-7, HT-29, PC-12 cancer and L-929 normal cell lines.Methods: The cells were incubated with different doses of NG-Ox and NG(50–1 000 mmol/L) for 24 h. The cell viability was assessed based on ATP cell viability assay.Intracellular accumulation of ROS was determined using the fluorescent probes 2070-dichlorodihydrofluorescin diacetate. Genotoxic effects were evaluated by alkaline single cell gel electrophoresis assay(comet assay) and, the apoptotic effect was evaluated by acridine orange staining at below the IC50 levels.Results: Both NG-Ox and NG exhibited cytotoxic, genotoxic and apoptotic effects and resulted in increased ROS values in a dose-dependent manner. The effects were more pronounced on cancer cell lines. The cytotoxic, genotoxic and apoptotic effects of NG-Ox were higher than that of NG in all cell lines. Significant correlations were observed between cell viability, DNA damage, apoptosis and ROS, in all cell lines exposed to either NG-Ox or NG.Conclusions: This study showed that both NG-Ox and NG possess cytotoxic, genotoxic and apoptotic activities through the production of ROS on cells, NG-Ox being the more effective one. Therefore, derived compound of NG might be used as antiproliferative agents for the treatment of cancer.
基金supported by the Key Research and Development Program of Shandong Province(Grant No.:2019GSF111001)the National Natural Science Foundation of China(Grant No.:21906096)+2 种基金the Youth Science Funds of the Shandong Academy of Sciences(Grant No.:2019QN009)the Youth Ph.D.Cooperation Funds of Qilu University of Technology(Shandong Academy of Sciences,Grant No.:2018BSHZ0029)the Program for Taishan Scholars of Shandong Province(Grant No.:tsqn202103099).
文摘Covalent organic nanospheres(CONs)were explored as a fiber coating for solid-phase microextraction of genotoxic impurities(GTIs)from active ingredients(AIs).CONs were synthesized by an easy solutionphase procedure at 25℃.The obtained nanospheres exhibited a high specific surface area,good thermostability,high acid and alkali resistance,and favorable crystallinity and porosity.Two types of GTIs,alkyl halides(1-iodooctane,1-chlorobenzene,1-bromododecane,1,2-dichlorobenzene,1-bromooctane,1-chlorohexane,and 1,8-dibromooctane)and sulfonate esters(methyl p-toluenesulfonate and ethyl ptoluenesulfonate),were chosen as target molecules for assessing the performance of the coating.The prepared coating achieved high enhancement factors(5097-9799)for the selected GTIs.The strong affinity between CONs and GTIs was tentatively attributed to π-π and hydrophobicity interactions,large surface area of the CONs,and size-matching of the materials.Combined with gas chromatography-mass spectrometry(GC-MS),the established analytical method detected the GTIs in capecitabine and imatinib mesylate samples over a wide linear range(0.2-200 ng/g)with a low detection limit(0.04-2.0 ng/g),satisfactory recovery(80.03%-109.5%),and high repeatability(6.20%-14.8%)and reproducibility(6.20%-14.1%).Therefore,the CON-coated fibers are promising alternatives for the sensitive detection of GTIs in AI samples.
基金Supported by The Intramural Research Program of the National Institutes of Health,Clinical Center
文摘Human pluripotent stem cells(h PSCs) represent heterogeneous populations, including induced pluripotent stem cells(i PSCs), endogenous plastic somatic cells, and embryonic stem cells(ESCs). Human ESCs are derived from the inner cell mass of the blastocyst, and they are characterized by the abilities to self-renew indefinitely, and to give rise to all cell types of embryonic lineage(pluripotency) under the guidance of the appropriate chemical, mechanical and environmental cues. The combination of these critical features is unique to h ESCs, and set them apart from other human cells. The expectations are high to utilize h ESCs for treating injuries and degenerative diseases; for modeling of complex illnesses and development; for screening and testing of pharmacological products; and for examining toxicity, mutagenicity, teratogenicity, and potential carcinogenic effects of a variety of environmental factors, including ionizing radiation(IR). Exposures to genotoxic stresses, such as background IR, are unavoidable; moreover, IR is widely used in diagnostic and therapeutic procedures in medicine on a routine basis. One of the key outcomes of cell exposures to IR is the change in gene expression, which may underlie the ultimate h ESCs fate after such a stress. However, gaps in our knowledge about basic biology of h ESCs impose a serious limitation to fully realize the potential of h ESCs in practice. The purpose of this review is to examine the available evidence of alterations in gene expression in human pluripotent stem cells after genotoxic stress, and to discuss strategies for future research in this important area.
基金funded by the National Natural Science Foundation of China(Nos.21103059,51136002 and 51076079)the China Key Technologies R&D Program(No.2012BAJ02B03)
文摘In the present study, a disposable electrochemical DNA nano-biosensor is proposed for the rapid detection of genotoxic compounds and bio-analysis of water pollution. The DNA nano-biosensor is prepared by immobilizing DNA on Au nanoparticles and a self-assembled monolayer of cysteamine modified Au electrode. The assembly processes of cysteamine, Au nanoparticles and DNA were characterized by cyclic voltammetry （CV）. The Au nanoparticles enhanced DNA immobilization resulting in an increased guanine signal. The interaction of the analyte with the immobilized DNA was measured through the variation of the electrochemical signal of guanine by square wave voltammetry （SWV）. The biosensor was able to detect the known genotoxic compounds： 2-anthramine, acridine orange and 2- naphthylamine with detection limits of 2, 3 and 50 nmol/L, respectively. The biosensor was also used to test actual water samples to evaluate the contamination level. Additionally, the comparison of results from the classical genotoxiciw bioassay has confirmed the applicability of the method for real samoles.
文摘Highly sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed for the simultaneous determination of five potential genotoxic impurities in ranolazine active pharmaceutical ingredient. Chromatographic separation achieved using Poroshell C18 PFP 150 × 3.0 mm 2.7 μ column and 0.1% formic acid in water as Mobile phase A and 0.1% formic acid in methanol as mobile phase B using gradient elution and a flow rate of 0.4 ml/min with a run time of 18 minutes. Mass spectrometric conditions were optimized using electrospray ionization in positive mode. Method shows excellent linearity from 0.05 - 5.0 ppm of the ranolazine test concentration for all the five impurities. The correlation coefficient was observed greater than 0.99. Satisfactory recoveries were observed for all the five impurities within the range of 102.9% - 112.3%. Method has been validated as per ICH recommended guidelines with a LOQ of 0.15 ppm achieved. The developed method was able to quantify all the five impurities at a concentration level of 1 ng/ml (0.5 ppm with respect to 2 mg/ml ranolazine).
文摘Nitrosamine impurities are potentially genotoxic which are considered under cohort of concern as per ICH M7 guidelines and need to be controlled at trace levels during quantification in drug substances and drug products for safe human consumption. Recent regulatory requirements also suggest the need to have highly sensitive analytical methods for the accurate quantification of Nitrosamine impurities. In this paper we have presented simple, rapid and ultra-sensitive LC-MS/MS method for six potential genotoxic nitrosamine impurities: N-Nitroso dimethyl amine (NDMA), N-Nitroso diethyl amine (NDEA), N-Nitroso Ethyl Iso propylamine (NEIPA), N-Nitroso-N-methyl-4-aminobutyric acid (NMBA) N-Nitroso diisopropylamino (NDIPA) and N-Nitroso dibutyl amine (NDBA) with a LOQ of 0.004 ppm. Chromatographic separation is achieved using Zorbax SB C18 150 × 3.0 mm, 3.5 μ column with 0.1% formic acid in water as mobile phase A and 0.1% formic acid in methanol as mobile phase B at a flow rate of 0.3 ml/min using gradient mode of elution at a total run time of 18 minutes. Six nitrosamine impurities are successfully ionized and quantified in positive mode of atmospheric pressure chemical ionization (APCI) using multiple reaction monitoring (MRM). Method validation is performed as per ICH guidelines evaluating the limit of quantification and detection and found to give good S/N ratios with good linearity range of 0.002 - 2 ppm with regression coefficient >0.99 for all the six nitrosamine impurities. Method recoveries are established using three-step sample preparation protocol and are found to be satisfactory within 80% - 120%. The method can be used routinely applied for the detection of Nitrosamines in Telmisartan at a concentration of 1.5 ng/ml (0.03 ppm with respect to telmisartan concentration of 50 mg/ml).
文摘Defining impurity profile is key element to ensure safe, efficacious and quality human drugs. Impurity profiling changed/transformed drastically over the years. Guidelines, specifications and requirements are evolving. Initially impurity profiling was based on simple methods later by degradation studies, then to understand drug strength and efficacy chiral impurities and stereo isomers were included followed by residual solvents, polymorphic forms, genotoxic impurity studies. Currently, elemental impurities are the latest addition. As per the GDUFA II guidelines to improve review efficiency and reduce review cycles, data requirements have changed. Based on recent guidance and review points, Impurity profiling has significant importance in ANDA filing and to ensure approval within 10 months (first cycle approval) which is an exiling aspect for industries to enter into the generic market quickly. Hence, Impurity profile is a key aspect scientifically, regulatory wise and commercially also. This is a review article on impurity profiling of Solid oral drug substances and products as per GDUFA II requirements the reference documents for the review are ICH guidance, relevant FDA GDUFA guidance and common industry practices.
基金The work was supported by Natural Science Foundation of China(Grant No. A20077023 and C40106012
文摘In this study, two common pollutants (benzo[a]pyrene and methamidophos) in marine environment were tested by comet assay for their inducement of in vivo genotoxic effect to the blood cells of black porgy ( Acanthopagrus schlegeli). The fish was exposed to 2 μg/L of benzo[a]pyrene (BaP) and methamidophos, and their mixture. The assay was performed on whole blood at 2 h, 5 h, 24 h and 96 h exposure intervals. A signifi- cant increase in DNA damage was observed in each treatment with the pollutants. Additive effect of BaP and methamidophos was also found in the experiment. However, the decrease ratios of DNA damage for 5 h and 96 h exposure interals compared with 2 h and 24 h exposure ones, respectively, were noticed. This phenomenon may be explained by the function of repairing process via enzyme cytochrome P450 in the animal. Evidence of the genotoxicity of organophosphorus pesticides (OPs) and polynuclear aromatic hydrocarbons (PAHs) on marine fish are discussed in this paper.
文摘A series of bioassays such as sister chromatid exchange frequencies ( SCE.), chromosomal aberration ( CA ), micronuclel rate (MN) and cell-cycle delay have been used to detecting the genotoxic effect of cigarette smoke condensate (CSC) on human diploid cell 2BS strain. The results suggested that a higher SCE, ( 17. 0/ cell) was observed In 2BS cells treated with CSC at 100 μg/ml, as compared with 6. 9/cell of the background (P<0. 001). CA rate was significantly increased from 4% to 36% In cells treated with 10 μg/ml CSC (P< 0.001). MN rate varied from 9 -26‰ In cells treated with CSC compared to that of control (6‰). Meanwhile, the cell-cycle of cells was markedly delayed by CSC. The survival rate of 2BS cells declined to 59. 6% for treatment with CSC at 200 μg/ ml. There was a dose-effect response In SCE., CA, MN rate. We proposed that active oxygen might responsible for genotoxiclty of CSC on cells.
文摘Medicinal use of spices/herbs has been gradually increased in the developed countries, Zingiber officinale （Ginger） is known to possess potent antioxidant and anti inflammatory properties. Therefore, the aim of this study is to determine the possible anti-mutagenic effect of ginger against the genotoxic effect of anti-cancer drug Taxol 0.6 mg/kg. This study is conducted by using two types of cytogenetic studies in bone marrow cell of mal albino mice Mus musculus （average weight 25-30 g）. The animals were randomly distributed into six groups, each of 14 mi[ce, （GI） was given the solvent, （G2） treatment of the medical dose of Taxol drug, （G3） treatment of ginger, （G4） a pre-treatment of ginger prior to treatment of drug, （G5） a simultaneous treatment of ginger and treatment of drug, （G6） a post-treatment of ginger after treatment drug. The study results show that significant increase in total chromosomal aberrations and significant increase in the number of micronuclei were observed after treatment drug. The significant structural aberrations were in the form of end-to-end associations. The numerical chromosomal aberrations were endomitosis and polyploid. The results showed that the frequencies of chromosomal aberrations and micronuclei in ginger treated group were not significantly different from control. Simultaneous treatment of ginger was found to be effective in reducing the genotoxic effects induced by drug Taxol especially in the total number of the chromosomal aberrations and the number of micronuclei.
文摘A simple, rapid, and highly sensitive LC-MS/MS method has been developed for the simultaneous and trace level quantification of underivatized boronic acids in lumacaftor active pharmaceutical ingredient. Chromatographic separation of boronic acids and lumacaftor achieved using Agilent Poroshell HPH C18 150<span> </span><span>×</span><span> </span><span>4.6</span><span> </span><span>mm 2.7</span><span> </span><span>μ column with 0.1% ammonia in water as mobile phase A and 100%</span><span> </span><span>acetonitrile as mobile phase B at a flow rate of 0.25</span><span> </span><span>ml/min. Gradient elution</span><span> was</span><span> used with a total method run time of 14</span><span> </span><span>minutes. Boronic acids were successfully ionized and quantified without derivatization using electrospray ionization in negative mode using tandem quadrupole mass spectrometry in multiple reactions monitoring mode. Method validation </span><span>was </span><span>performed as per ICH guidelines with good linearity over the concentration range of 0.05 ppm to 5 ppm of Lumacaftor test concentration for both the boronic acids with a correlation coefficient of >0.99.</span><span> </span><span>Recoveries were found good at different concentration levels and within the range of 80</span><span>% </span><span>-</span><span> </span><span>120%.</span><span> </span><span>The developed method can be successfully used for the routine quantification of boronic acids at a concentration level of 20</span><span> </span><span>ng/ml (1</span><span> </span><span>ppm with respect to 20</span><span> </span><span>mg/ml lumacaftor).</span>
文摘The genotoxic activities of effluents from drainage water treatment plants were examined by using the novel <em>umu</em> tester strain NM8001, which lacks <em>MutMst</em> genes. To enhance the sensitivity of the LacZ assay, a BugBuster mix protein extraction reagent and TokyoGreen-<em>β</em> Gal for a fluorescence-galactosidase substrate were applied. Of the 24 sampling locations present in Kanagawa prefecture, Japan, water extracts from nine sampling points showed apparent genotoxic activities without metabolic activation. In contrast, water extracts from the upper sites of these water treatment plants did not show any significant genotoxic activities. The selected samples with genotoxic activity did not show significant mutagenicity toward Ames strains TA98 and TA100. Genotoxicity was also well correlated with the activity of a classical <em>umu </em>strain of TA1535/pSK1002;these findings indicate that the genotoxicity induced by oxidative damage was not a significant component of the genotoxicity.
文摘The no-observed-effect level (NOEL) in a study of carcinogenicity for compounds that are both genotoxic and carcinogenic represents the limit of detection in that bioassay, rather than an estimate of a possible threshold. Therefore, for those genotoxic and carcinogenic contaminants (e.g. acrylamides, PAHs, etc.) in foods it is not possible to develop health-based guidance values (e.g. ADI or PTWI) using the traditional NOEL and safety/uncertainty factors.
文摘4-hydrazinobenzoic acid has been highlighted as one of the potential genotoxic impurities (PGIs). A sensitive HPLC method was developed and validated for the determination of 4-hydrazino benzoic acid in Deferasirox Tablet. HPLC method on Zorbax SB C18 column (250 × 4.6 mm i.d.), 5 μm, with UV Detector was used. The proposed method was cost effective, specific, linear, accurate, rugged and precise. The calibration curves showed good linearity over the concentration range of 0.5 μg/ml to 1.5 μg/ml with respect to the sample. The correlation coefficient was 0.999. Excellent recoveries of 102% were obtained at the level 0.5 μg/ml.
文摘With the ever increasing complexity of active pharmaceutical ingredient (API) preparations, more potential genotoxic impurities (PGI’s) are being observed. It is thus necessary to determine if these PGI’s are present in the final API’s, and if they are present, to ensure the levels are acceptable for any clinical uses. For PGI’s that have authentic standards available, quantitation can be accomplished in a straightforward manner. However, for PGI’s that are expected to form through rearrangements or side reactions, authentic standards may not be readily available, significantly complicating the analysis. In this study we describe a surrogate standard approach for quantifying PGI’s that allows for relative response factor calculations of PGI species utilizing both gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS).
文摘Background: In oral and maxillofacial surgery, synthetic bone grafts are most widely used as bone substitutes, due to the limited sources of autologous bone. The aim of this study was to examine the influence of three different synthetic bone grafts (Cerasorb, Fortoss and Perioglass) on sisters chromatide exchanges (SCEs) in peripheral lymphocytes. Materials and Methods: Peripheral blood samples taken from 68 patients (45 females and 23 males), who underwent oral surgery procedures, such as apical resection, cyst enucleation or periodontal curretage, were obtained for SCE a day before and two months after the surgeries. A control group included 30 patients, while the study group was made of the patients who underwent bone grafting with Cerasorb? (11 patients), Fortoss? VITAL (10 patients) or Perioglass? (17 patients). Results: Comparing with the results of the study group before and after the treatment, it was concluded that the results were statistically significant (p = 0.001). In the Perioglass? subgroup, a greater statistical significance (p = 0.003) was noted, than that in either the Cerasorb? (p = 0.620) or Fortoss? (p = 0.210) subgroups, in which there was no statistical significance. Conclusions: Although further investigations may be necessary, our results suggest that the synthetic bone grafts might have an influence on SCE in peripheral lymphocytes.
基金the Fundaç~ao de Amparoa Ci^encia e Tecnologia de Pernambuco e FACEPE,Núcleo de Química Analítica Avançada de Pernambuco-NUQAAPE(FACEPE,process APQ-0346-1.06/14)FACEPE(process APQ-0947-3.06/22)and FADE/UFPE.
文摘Population growth and the increase in the consumption of different pharmaceuticals combined with the insufficiency in the removal of these compounds by conventional treatments have contributed to the increase in the detection of these contaminants in aquatic matrices.Aiming to contribute in solving this problem,this promoted the degradation of a mixture of the drugs lamivudine and zidovudine in different matrices(aqueous solution and synthetic effluent)using the heterogeneous photo-Fenton process applying pyrite as a catalyst and artificial solar radiation.At the end of the treatment,degradations greater than 99%were found for zidovudine in both matrices studied,while for lamivudine,97%and 94%degradations were obtained for aqueous solution and synthetic effluent,in that order.In the investigation of toxic effects using Biomphalaria glabrata molluscs,embryotoxicity tests showed embryonic lethality in 100%of individuals for all samples.Acute toxicity tests on adult molluscs resulted in mortality rates of 100%(aqueous solution after treatment)and 50%(synthetic effluent after treatment).Thus,to investigate cellular changes,genotoxicity analyses were carried out,and different degrees of DNA damage were observed,however,the highest level of damage to this organism was not observed.Therefore,B.glabrata demonstrated to be sensitive to toxic effects at the concentrations present in the matrices studied,providing evidence to predict the ecotoxicological potential of samples when released into aquatic ecosystems.
文摘Green synthesis of silver nanoparticles(NPs)by green route approaches has advantages over conventional methods.In green synthesis,we use eco-friendly plant extracts contain secondary metabolites and bioactive components,proteins that act as both reducing and capping agents,form stable and shape-controlled green silver nanoparticles.The current study deals with the synthesis of silver nanoparticles using the aqueous latex extract of Allamanda cathartica.The green silver nanoparticles are characterized by using different spectroscopic methods like ultra violet-visible spectroscopy(UV-Vis),Fourier transform-infrared spectroscopy(FTIR),transmission electron microscope(TEM),scanning electron microscope(SEM)and X-ray diffraction(XRD).Results indicated that the crystalline natured particles were spherical shaped with an average of 35 nm in size,and that the stability of silver nanoparticles was due to its high negative zeta potential of–27.6 mV.The current study also revealed that green silver nanoparticles had very good genotoxic and cytotoxic activity in peripheral blood mononuclear cells(PBMCs).Leukemia leads to the development of high numbers of white blood cells,which is one of the major types of cancers that affect children.Many of the chemicals used for the treatment produce remarkable side effects.To overcome this problem,we made an attempt to see the efficacy of latex green silver nanoparticle on peripheral blood mononuclear cells and deoxyribonucleic acid fragmentation,which leads to the development of future therapeutic drugs.
