An in vitro liver model - assessing oxidative stress and genotoxicity following exposure of hepatocytes to a panel of engineered nanomaterials

ABSTRACT: BACKGROUND: Following exposure via inhalation, intratracheal instillation or ingestion some nanomaterials(NM) have been shown to translocate to the liver. Since oxidative stress has been implicatedas a possible mechanism for NM toxicity this study aimed to investigate the effects of variousmaterials (five titanium dioxide (TiO2), two zinc oxide (ZnO), two multi-walled carbonnanotubes (MWCNT) and one silver (Ag) NM) on oxidative responses of C3A cell line as amodel for potential detrimental properties of nanomaterials on the liver. RESULTS: We noted a dose dependant decrease in the cellular glutathione content following exposure ofthe C3A cells to Ag, the ZnO and the MWCNTs. Intracellular ROS levels were alsomeasured and shown to increase significantly following exposure of the C3A to the lowtoxicity NMs (MWCNT and TiO2). The antioxidant Trolox in part prevented the detrimentaleffect of NMs on cell viability, and decreased the NM induced IL8 production after exposureto all but the Ag particulate. Following 4 hr exposure of the C3A cells to sub-lethal levels ofthe NMs, the largest amount of DNA damage was induced by two of the TiO2 samples (7 nmand the positively charged 10 nm particles). CONCLUSIONS: All ten NMs exhibited effects on the hepatocyte cell line that were at least in partROS/oxidative stress mediated. These effects included mild genotoxicity and IL8 productionfor all NM except the Ag possibly due to its highly cytotoxic nature.     

氟氯菊酯对大型蚤的亚慢性毒性及其恢复实验

本实验研究了氟氯菊酯对大型蚤母代F0的亚慢性实验,及其两代子代(第一胎和第三胎:F1(1st)和F1(3rd))在没有农药的环境中生存21d的恢复实验,并且用存活期、体长、繁殖(每个母蚤产幼蚤数、第一次怀卵和产卵的天数、每个母蚤的胎数)和内禀增长率(r)等参数来进行评价。在恢复实验中,第一胎大型蚤的每只母蚤产幼蚤数和体长这两个参数在恢复实验中仍然受到明显的抑制,第三胎大型蚤在没有农药的环境中生存了21d后的各个参数基本上能恢复的与对照组没有显著区别了,但是在0.5和0.75μg/kg的浓度中生存后的母蚤所产的第三胎在体长参数上还没有完全的恢复(P<0.05)。     

Impact of an Engineered Copper-Titanium Dioxide Nanocomposite and Parent Substrates on the Bacteria Viability,Antioxidant Enzymes and Fatty Acid Profiling

Due to the systematic increase in the production of nanomaterials (NMs) and their applications in many areas of life, issues associated with their toxicity are inevitable. In particular, the performance of heterogeneous NMs, such as nanocomposites (NCs), is unpredictable as they may inherit the properties of their individual components. Therefore, the purpose of this work was to assess the biological activity of newly synthesized Cu/TiO₂-NC and the parent nanoparticle substrates Cu-NPs and TiO₂-NPs on the bacterial viability, antioxidant potential and fatty acid composition of the reference Escherichia coli and Bacillus subtilis strains. Based on the toxicological parameters, it was found that B. subtilis was more sensitive to NMs than E. coli. Furthermore, Cu/TiO₂-NC and Cu-NPs had an opposite effect on both strains, while TiO₂-NPs had a comparable mode of action. Simultaneously, the tested strains exhibited varied responses of the antioxidant enzymes after exposure to the NMs, with Cu-NPs having the strongest impact on their activity. The most considerable alternations in the fatty acid profiles were found after the bacteria were exposed to Cu/TiO₂-NC and Cu-NPs. Microscopic images indicated distinct interactions of the NMs with the bacterial outer layers, especially in regard to B. subtilis. Cu/TiO₂-NC generally proved to have less distinctive antimicrobial properties on B. subtilis than E. coli compared to its parent components. Presumably, the biocidal effects of the tested NMs can be attributed to the induction of oxidative stress, the release of metal ions and specific electrochemical interactions with the bacterial cells.     

Effects of Prenatal Phthalate Exposure and Childhood Exercise on Maternal Behaviors in Female Rats at Postpartum: A Role of Oxtr Methylation in the Hypothalamus

Both the detrimental effect of prenatal exposure to di-(2-ethylhexyl)-phthalate (DEHP) and the beneficial effects of physical exercise on brain functions have been reported. The oxytocin pathway has been implicated in the onset of maternal behaviors. Epigenetic modification of the oxytocin receptor gene (OXTR) through DNA methylation has been associated with the pathogenesis of neuropsychiatric disorders. The purpose of this study was to investigate the effects of prenatal DEHP exposure on oxytocin-regulated maternal behaviors and to examine the protective effect of exercise. Pregnant rats (F0) were fed with vehicle or DEHP during gestation and the offspring females (F1) were assessed for their maternal behaviors by pup retrieval test at postpartum. The results showed that reduced pup retrieval activities without significant alteration of stress responses were observed in the prenatally DEHP-exposed females. Prenatal DEHP exposure decreased the expressions of oxytocin, Oxtr mRNA, and oxytocin receptor, and increased Oxtr methylation in the hypothalamus of postpartum female rats. There were no significant effects of exercise on behavioral, biochemical, and epigenetic measurements. These results suggest that prenatal DEHP exposure has a long-term adverse effect on maternal behaviors; Oxtr hyper-methylation may be a potential epigenetic mechanism for this alteration, which cannot be prevented by physical exercise during childhood.     

Phase-transformation synthesis of Li codoped ZrO2: Eu3+ nanomaterials: Characterization,photocatalytic, luminescent behaviour and latent fingerprint development

In recent years, key study was designing nanomaterials (NMs) with tunable properties in order to obtain high functional materials. For this purpose, an attempt with great effort has been put forward to synthesize ZrO₂:Eu:Li (1–11 mol%) NM with varying concentration of Li⁺ ion that can have a control over phase, size, morphology, crystallinity, band gap and surface chemistry. More importantly, addition of Li as codopant influences the phase and crystallinity change with the change in concentration. However important challenges faced by this work was to understand the phase change, crystallinity, structural change and photocatalytic activity which has to be still explored. The synthesized NMs possess mixed phase and cubic phase with change in concentration of codopant which may be attributed to presence of defect states, micro strain, distortion of lattice. The energy band gap was found to decrease for 7 mol% NM attributed to the change in the phase. Porous morphology with variation in pore length was observed. Enhanced luminescence intensity with intense orange red emissions consistent to ⁵D₀→⁷F_j (j = 0 to 4) intra configurational f-f transitions was observed for ZrO₂:Eu:Li⁺(1–11 mol%) nanophosphor excited at 394 nm. The visualization of latent fingerprints using ZrO₂: Eu: Li⁺_7mol% nanophosphor on several surfaces, the powder dusting technique was adopted. The enhanced fingerprint under UV light provides well resolved ridge patterns for the identification of individual latent finger prints using ZrO₂:Eu:Li (7 mol%) with clear resolution. Lower charge transfer resistance with enhanced photocatalytic activity for decolourization of Rhodamine B with high pore length to allow multiple reflections under UV light irradiation for 7 mol% NM with reduced band gap and optimum luminescence intensity was observed. Hence, the synthesized ZrO₂:Eu:Li (7 mol%) can be employed in forensic science towards latent fingerprint development, as a photocatalyst for environmental remediation and as luminescent material in display applications.     

NanoTIO2 (UV-Titan) does not induce ESTR mutations in the germline of prenatally exposed female mice

Background

Particulate air pollution has been linked to an increased risk of cardiovascular disease and cancer. Animal studies have shown that inhalation of air particulates induces mutations in the male germline. Expanded simple tandem repeat (ESTR) loci in mice are sensitive markers of mutagenic effects on male germ cells resulting from environmental exposures; however, female germ cells have received little attention. Oocytes may be vulnerable during stages of active cell division (e.g., during fetal development). Accordingly, an increase in germline ESTR mutations in female mice prenatally exposed to radiation has previously been reported. Here we investigate the effects of nanoparticles on the female germline. Since pulmonary exposure to nanosized titanium dioxide (nanoTiO₂) produces a long-lasting inflammatory response in mice, it was chosen for the present study.

Findings

Pregnant C57BL/6 mice were exposed by whole-body inhalation to the nanoTiO₂ UV-Titan L181 (~42.4?mg UV-Titan/m³) or filtered clean air on gestation days (GD) 8?C18. Female C57BL/6?F1 offspring were raised to maturity and mated with unexposed CBA males. The F2 descendents were collected and ESTR germline mutation rates in this generation were estimated from full pedigrees (mother, father, offspring) of F1 female mice (192 UV-Titan-exposed F2 offspring and 164?F2 controls). ESTR mutation rates of 0.029 (maternal allele) and 0.047 (paternal allele) in UV-Titan-exposed F2 offspring were not statistically different from those of F2 controls: 0.037 (maternal allele) and 0.061 (paternal allele).

Conclusions

We found no evidence for increased ESTR mutation rates in F1 females exposed in utero to UV-Titan nanoparticles from GD8-18 relative to control females.     

Controlled human wood smoke exposure: oxidative stress, inflammation and microvascular function

Background

Due to the rising use of nanomaterials (NMs), there is concern that NMs induce undesirable biological effects because of their unique physicochemical properties. Recently, we reported that amorphous silica nanoparticles (nSPs), which are one of the most widely used NMs, can penetrate the skin barrier and induce various biological effects, including an immune-modulating effect. Thus, it should be clarified whether nSPs can be a risk factor for the aggravation of skin immune diseases. Thus, in this study, we investigated the relationship between the size of SPs and adjuvant activity using a model for atopic dermatitis.

Results

We investigated the effects of nSPs on the AD induced by intradermaly injected-mite antigen Dermatophagoides pteronyssinus (Dp) in NC/Nga mice. Ear thickness measurements and histopathological analysis revealed that a combined injection of amorphous silica particles (SPs) and Dp induced aggravation of AD in an SP size-dependent manner compared to that of Dp alone. In particular, aggravation was observed remarkably in nSP-injected groups. Furthermore, these effects were correlated with the excessive induction of total IgE and a stronger systemic Th2 response. We demonstrated that these results are associated with the induction of IL-18 and thymic stromal lymphopoietin (TSLP) in the skin lesions.

Conclusions

A particle size reduction in silica particles enhanced IL-18 and TSLP production, which leads to systemic Th2 response and aggravation of AD-like skin lesions as induced by Dp antigen treatment. We believe that appropriate regulation of nanoparticle physicochemical properties, including sizes, is a critical determinant for the design of safer forms of NMs.     

Transgenerational Effects of Di(2-Ethylhexyl) Phthalate on Anogenital Distance,Sperm Functions and DNA Methylation in Rat Offspring

Di(2-ethylhexyl) phthalate (DEHP) is widely used as a plasticizer in the manufacture of polyvinylchloride plastics and has been associated with concerns regarding male reproductive toxicity. In this study, we hypothesized that maternal exposure to DEHP induces transgenerational inheritance of adult-onset adverse reproductive outcomes through the male germline in the F1, F2, and F3 generations of male offspring. Pregnant rats were treated with 5 or 500 mg of DEHP/kg/day through gavage from gestation day 0 to birth. The offspring body weight, anogenital distance (AGD), anogenital index (AGI), sperm count, motility, and DNA fragmentation index (DFI) were measured for all generations. Methyl-CpG binding domain sequencing was performed to analyze sperm DNA methylation status in the F3. DEHP exposure at 500 mg/kg affected AGD, AGI, sperm count, mean DFI, and %DFI in the F1; AGD, sperm count, and mean DFI in the F2; and AGD, AGI, mean DFI, and %DFI in the F3. DEHP exposure at 5 mg/kg affected AGD, AGI, sperm count, and %DFI in the F1; sperm count in the F2; and AGD and AGI in F3. Compared with the control group, 15 and 45 differentially hypermethylated genes were identified in the groups administered 5 mg/kg and 500 mg/kg DEHP, respectively. Moreover, 130 and 6 differentially hypomethylated genes were observed in the groups administered 5 mg/kg and 500 mg/kg DEHP. Overall, these results demonstrated that prenatal exposure to DEHP caused transgenerational epigenetic effects, which may explain the observed phenotypic changes in the male reproductive system.     

Defense Tradeoffs in Fleshy Fruits: Effects of Resource Variation on Growth, Reproduction, and Fruit Secondary Chemistry in Solanum carolinense

A set of clones of 10 maternal plants was grown for three successive years (1998-2000) under two nitrogen treatments and two water treatments. Path analysis revealed strong direct and indirect effects of nitrogen treatment on growth and reproduction, but fruit morphological and chemical variables were not strongly affected. Fruit pulp chemistry varied only slightly across treatments despite the large differences in growth and reproduction associated with resource variation. Leaf and ripe fruit chemical contents were not significantly correlated across treatments, and maternal plants, and leaf chemical variables did not help explain fruit chemical variation when included as covariates in ANCOVA analyses, suggesting no physiological constraints of leaf chemistry on ripe fruit chemistry. Results suggest that, while maternal plants may vary somewhat in fruit chemistry, and fruit chemistry may vary somewhat depending upon environmental conditions, levels of primary and secondary metabolites within fruits are not best explained by supply-side hypotheses. Ripe fruit chemistry remained relatively constant in the face of drastically changing resource levels, suggesting an adaptive function and supporting the Defense Tradeoff hypothesis. Fruit quality, both in terms of nutritional make-up and putative defensive properties, was maintained despite strong effects on plant growth and reproduction. Because glycoalkaloids are general defense compounds, we conclude that ripe fruit chemistry most likely reflects a balance between selection for attraction of seed dispersers and defense against pests and pathogens.     

Studies on preparations and pH/redox responsiveness of zwitterionic nanomicelles of poly[lysine-co-N,N-bis(acryloyl)cystamine-co-dodecylamine]

Poly[L-lysine-co-N,N-bis(acryloyl) cystamine-co-dodecylamine] nanomicelles (NMs) were synthesized through Michael addition terpolymerization in one pot. The NMs showed spheric morphology and uniform size distributions. The NMs had excellent nonspecific protein adsorption ability at pH 7.4. Doxorubicin (DOX) was loaded in the NMs for the investigation of controlled release. The drug delivery results showed that the DOX-loaded NMs displayed apparent pH and reduction sensitivities in response to the environment of tumor cells due to the existence of carboxyl groups, amino groups, and disulfide bonds in the NMs. The NMs were biocompatible, biodegradable, and could be potentially used as drug vehicles in controlled release.     

Effect of cell density on cytotoxicity of ether lipid analogues in variants of B16 murine melanoma

Ether lipids are defined here as analogues of naturally occurring lysophosphatidylcholines with cytotoxic activity against neoplastic cells. The activity of 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine (ET18OMe) and 3-hexadecylmercapto-2-methoxymethyl-propyl-1-phosphocholine (Ilmofosine^®) (BM 41.440) was tested in variants of B16 murine melanoma, grown in adhesion cultures (B16F1 with low metastatic potential; B16F10 and B16BL6 with high metastatic potential). Cytotoxicity was evaluated by counting the cells that survived after 24h of drug exposure. Cholesterol, sphingomyelin, total phospholipid and phosphatidylcholine levels were determined. After 24 h of drug exposure, cultures of the B16BL6 variant contained a larger number of cells, especially when high drug concentrations (100–250 μM) were used, than cultures of the B16F1 and B16F10 variants. The sensitivity of ET18OMe of the three variants was evaluated at different cell densities (at each density the dose was equalized per number of cells/well; 0.1 μmol/10⁶ cells/well). In B16F1 and B16F10 cultures the dose-response curve was not affected by the number of cells/well, while in B16BL6 no more than 20% of the cells were killed at all cell densities measured. A linear relationship was noted between cell density and cholesterol/phospholipid and sphingomyelin/phosphatidylcholine ratios in the resistant variant B16BL6, confirming that lipid composition modulates the cytotoxic activity of ether lipids.     

Effects of Electromagnetic Waves with LTE and 5G Bandwidth on the Skin Pigmentation In Vitro

With the rapid growth of wireless communication devices, the influences of electromagnetic fields (EMF) on human health are gathering increasing attention. Since the skin is the largest organ of the body and is located at the outermost layer, it is considered a major target for the health effects of EMF. Skin pigmentation represents one of the most frequent symptoms caused by various non-ionizing radiations, including ultraviolet radiation, blue light, infrared, and extremely low frequency (ELF). Here, we investigated the effects of EMFs with long-term evolution (LTE, 1.762 GHz) and 5G (28 GHz) bandwidth on skin pigmentation in vitro. Murine and Human melanoma cells (B16F10 and MNT-1) were exposed to either LTE or 5G for 4 h per day, which is considered the upper bound of average smartphone use time. It was shown that neither LTE nor 5G exposure induced significant effects on cell viability or pigmentation. The dendrites of MNT-1 were neither lengthened nor regressed after EMF exposure. Skin pigmentation effects of EMFs were further examined in the human keratinocyte cell line (MNT-1-HaCaT) co-culture system, which confirmed the absence of significant hyper-pigmentation effects of LTE and 5G EMFs. Lastly, MelanoDerm™, a 3D pigmented human epidermis model, was irradiated with LTE (1.762 GHz) or 5G (28 GHz), and image analysis and special staining were performed. No changes in the brightness of MelanoDerm™ tissues were observed in LTE- or 5G-exposed tissues, except for only minimal changes in the size of melanocytes. Collectively, these results imply that exposure to LTE and 5G EMFs may not affect melanin synthesis or skin pigmentation under normal smartphone use condition.     

Deciphering Differential Life Stage Radioinduced Reproductive Decline in Caenorhabditis elegans through Lipid Analysis

Wildlife is chronically exposed to various sources of ionizing radiations, both environmental or anthropic, due to nuclear energy use, which can induce several defects in organisms. In invertebrates, reproduction, which directly impacts population dynamics, has been found to be the most radiosensitive endpoint. Understanding the underlying molecular pathways inducing this reproduction decrease can help in predicting the effects at larger scales (i.e., population). In this study, we used a life stage dependent approach in order to better understand the molecular determinants of reproduction decrease in the roundworm C. elegans. Worms were chronically exposed to 50 mGy·h⁻¹ external gamma ionizing radiations throughout different developmental periods (namely embryogenesis, gametogenesis, and full development). Then, in addition to reproduction parameters, we performed a wide analysis of lipids (different class and fatty acid via FAMES), which are both important signaling molecules for reproduction and molecular targets of oxidative stress. Our results showed that reproductive defects are life stage dependent, that lipids are differently misregulated according to the considered exposure (e.g., upon embryogenesis and full development) and do not fully explain radiation induced reproductive defects. Finally, our results enable us to propose a conceptual model of lipid signaling after radiation stress in which both the soma and the germline participate.     

Epigenetic effects of environmental chemicals bisphenol a and phthalates

The epigenetic effects on DNA methylation, histone modification, and expression of non-coding RNAs (including microRNAs) of environmental chemicals such as bisphenol A (BPA) and phthalates have expanded our understanding of the etiology of human complex diseases such as cancers and diabetes. Multiple lines of evidence from in vitro and in vivo models have established that epigenetic modifications caused by in utero exposure to environmental toxicants can induce alterations in gene expression that may persist throughout life. Epigenetics is an important mechanism in the ability of environmental chemicals to influence health and disease, and BPA and phthalates are epigenetically toxic. The epigenetic effect of BPA was clearly demonstrated in viable yellow mice by decreasing CpG methylation upstream of the Agouti gene, and the hypomethylating effect of BPA was prevented by maternal dietary supplementation with a methyl donor like folic acid or the phytoestrogen genistein. Histone H3 was found to be trimethylated at lysine 27 by BPA effect on EZH2 in a human breast cancer cell line and mice. BPA exposure of human placental cell lines has been shown to alter microRNA expression levels, and specifically, miR-146a was strongly induced by BPA treatment. In human breast cancer MCF7 cells, treatment with the phthalate BBP led to demethylation of estrogen receptor (ESR1) promoter-associated CpG islands, indicating that altered ESR1 mRNA expression by BBP is due to aberrant DNA methylation. Maternal exposure to phthalate DEHP was also shown to increase DNA methylation and expression levels of DNA methyltransferases in mouse testis. Further, some epigenetic effects of BPA and phthalates in female rats were found to be transgenerational. Finally, the available new technologies for global analysis of epigenetic alterations will provide insight into the extent and patterns of alterations between human normal and diseased tissues. In vitro models such as human embryonic stem cells may be extremely useful in bettering the understanding of epigenetic effects on human development, health and disease, because the formation of embryoid bodies in vitro is very similar to the early stage of embryogenesis.     

Effect of Nanoparticles Exposure on Fractional Exhaled Nitric Oxide (FENO) in Workers Exposed to Nanomaterials

Fractional exhaled nitric oxide (FENO) measurement is a useful diagnostic test of airway inflammation. However, there have been few studies of FENO in workers exposed to nanomaterials. The purpose of this study was to examine the effect of nanoparticle (NP) exposure on FENO and to assess whether the FENO is increased in workers exposed to nanomaterials (NM). In this study, both exposed workers and non-exposed controls were recruited from NM handling plants in Taiwan. A total of 437 subjects (exposed group = 241, non-exposed group = 196) completed the FENO and spirometric measurements from 2009–2011. The authors used a control-banding (CB) matrix to categorize the risk level of each participant. In a multivariate linear regression analysis, this study found a significant association between risk level 2 of NP exposure and FENO. Furthermore, asthma, allergic rhinitis, peak expiratory flow rate (PEFR), and NF-κB were also significantly associated with FENO. When the multivariate logistic regression model was adjusted for confounders, nano-TiO₂ in all of the NM exposed categories had a significantly increased risk in FENO > 35 ppb. This study found associations between the risk level of NP exposure and FENO (particularly noteworthy for Nano-TiO₂). Monitoring FENO in the lung could open up a window into the role nitric oxide (NO) may play in pathogenesis.     

Aflatoxin Exposure during Early Life Is Associated with Differential DNA Methylation in Two-Year-Old Gambian Children

Background: DNA methylation is an epigenetic control mechanism that may be altered by environmental exposures. We have previously reported that in utero exposure to the mycotoxin and liver carcinogen aflatoxin B1 from the maternal diet, as measured using biomarkers in the mothers’ blood, was associated with differential DNA methylation in white blood cells of 6-month-old infants from The Gambia. Methods: Here we examined aflatoxin B1-associated differential DNA methylation in white blood cells of 24-month-old children from the same population (n = 244), in relation to the child’s dietary exposure assessed using aflatoxin albumin biomarkers in blood samples collected at 6, 12 and 18 months of age. HM450 BeadChip arrays were used to assess DNA methylation, with data compared to aflatoxin albumin adduct levels using two approaches; a continuous model comparing aflatoxin adducts measured in samples collected at 18 months to DNA methylation at 24 months, and a categorical time-dose model that took into account aflatoxin adduct levels at 6, 12 and 18 months, for comparison to DNA methylation at 24 months. Results: Geometric mean (95% confidence intervals) for aflatoxin albumin levels were 3.78 (3.29, 4.34) at 6 months, 25.1 (21.67, 29.13) at 12 months and 49.48 (43.34, 56.49) at 18 months of age. A number of differentially methylated CpG positions and regions were associated with aflatoxin exposure, some of which affected gene expression. Pathway analysis highlighted effects on genes involved with with inflammatory, signalling and growth pathways. Conclusions: This study provides further evidence that exposure to aflatoxin in early childhood may impact on DNA methylation.     

The bacterial battery and the effect of different exposure conditions on biofilm properties

A galvanic cell with pure copper and Al 2024 and Shewanella oneidensis MR-1 in Luria Bertani (LB) medium and a control cell, which did not contain the bacteria, have been tested. Potentiodynamic polarization experiments were used to determine cell voltage (V)-current (i) and power (P)-i curves as a function of time. The results for the cell without bacteria demonstrated that the maximum power output was obtained in the first day and dropped drastically in the following days. For the cell with MR-1 the power output of the cell increased slowly with time for around 100 days and remained at similar values for another 100 days. For pure Cu significant changes of the impedance spectra were found in the presence of MR-1. The impedance spectra resembled those usually observed for polymer-coated metals. In order to investigate this phenomenon in more detail, the effects of the different exposure conditions on the electrochemical behavior of copper were evaluated in LB containing MR-1. A second time constant was observed in the impedance spectra of copper that was partially immersed in the test solution (cell B). Complete immersion of copper in the electrolyte (no air/liquid interface) or deaeration of cell B resulted in one-time-constant spectra that are typical of those found for passive metals. Excellent corrosion protection was provided by MR-1 regardless of exposure condition. The corrosion potential increased with time for the Cu electrode exposed in cell B, while it decreased for the other two exposure conditions.     

Effects of surface curvature and surface characteristics of carbon-based nanomaterials on the adsorption and activity of acetylcholinesterase

Carbon-based nanomaterials (NM) are promising candidates for a myriad of applications ranging from drug delivery to biosensing platforms. In the physiological environment, proteins can be adsorbed onto the surface of NM that can alter their structure and function. Little is known of the effect of NM on larger proteins and enzymes and an attempt has been made in this study to investigate the effect of carbon-based NM such as carbon black (CB), graphene oxide (GO) and fullerene (C₆₀) on the adsorption and activity of acetylcholinesterase (AChE), a key enzyme present in brain, blood and nervous system and a suitable neurotoxicity biomarker. Experimental and computational results showed that all the carbon-based NM tested adsorb AChE but they have different effects on the catalytic activity of the enzyme. The most efficient AChE inhibitor is CB. In contrast, AChE adsorbed on the GO surface retains its native conformation and most of its activity. As compared to GO and CB, C₆₀ was found to be an inefficient adsorbent of AChE. The distinctive adsorption pattern of NM and their inhibitory potential could be related to the surface characteristics of NM. Our studies also demonstrate the potential of GO as a substrate for immobilization of AChE.     

Modelling the human immune response: performance of a 1011 human antibody repertoire against a broad panel of therapeutically relevant antigens

A large 1.29 x 10(11) antibody fragment library, based upon variable (V) genes isolated from human B-cells from 160 donors has been constructed and its performance measured against a panel of 28 different clinically relevant antigens. Over 5000 different target-specific antibodies were isolated to the 28 antigens with 3340 identified as modulating the biological function (e.g. antagonism, agonism) of the target antigen. This represents an average of approximately 120 different functionally active antibodies per target. Analysis of a sample of >800 antibodies from the unselected library indicates V gene usage is representative of the human immune system with no strong bias towards any particular V(H)-V(L) pairing. Germline diversity is broad with 45/49 functional V(H) germlines and 28/30 V(lambda) and 30/35 V(kappa) light-chain germlines represented in the sample. The number of functional V(H) germlines and V(kappa) light-chain germlines present is increased to 48/49 and 31/35, respectively, when selected V gene usage is included in the analysis. However, following selection on the antigen panel, V(H)1-V(lambda)1 germline family pairings are preferentially enriched and represent a remarkable 25% of the antigen-specific selected repertoire.