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N-Nitrosamine impurities, including nitrosamine drug substance-related impurities (NDSRIs), have challenged pharmaceutical industry and regulators alike and affected the global drug supply over the past 5 years. Nitrosamines are a class of known carcinogens, but NDSRIs have posed additional challenges as many lack empirical data to establish acceptable intake (AI) limits. Read-across analysis from surrogates has been used to identify AI limits in some cases; however, this approach is limited by the availability of robustly-tested surrogates matching the structural features of NDSRIs, which usually contain a diverse array of functional groups. Furthermore, the absence of a surrogate has resulted in conservative AI limits in some cases, posing practical challenges for impurity control. Therefore, a new framework for determining recommended AI limits was urgently needed. Here, the Carcinogenic Potency Categorization Approach (CPCA) and its supporting scientific rationale are presented. The CPCA is a rapidly-applied structure-activity relationship-based method that assigns a nitrosamine to 1 of 5 categories, each with a corresponding AI limit, reflecting predicted carcinogenic potency. The CPCA considers the number and distribution of α-hydrogens at the N-nitroso center and other activating and deactivating structural features of a nitrosamine that affect the α-hydroxylation metabolic activation pathway of carcinogenesis. The CPCA has been adopted internationally by several drug regulatory authorities as a simplified approach and a starting point to determine recommended AI limits for nitrosamines without the need for compound-specific empirical data., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Published by Elsevier Inc.)

The use of advanced electroactive catalysts enhances the performance of electrochemical biosensors in real-time biomonitoring and has received much attention owing to its excellent physicochemical and electrochemical possessions. In this work, a novel biosensor was developed based on the electrocatalytic activity of functionalized vanadium carbide (VC) material, including VC@ruthenium (Ru), VC@Ru-polyaniline nanoparticles (VC@Ru-PANI-NPs) as non-enzymatic nanocarriers for the fabrication of modified screen-printed electrode (SPE) to detect acetaminophen in human blood. As-prepared materials were characterized using SEM, TEM, XRD, and XPS techniques. Biosensing was carried out using cyclic voltammetry and differential pulse voltammetry techniques and has revealed imperative electrocatalytic activity. A quasi-reversible redox method of the over-potential of acetaminophen increased considerably compared with that at the modified electrode and the bare SPE. The excellent electrocatalytic behaviour of VC@Ru-PANI-NPs/SPE is attributed to its distinctive chemical and physical properties, including rapid electron transfer, striking ᴫ-ᴫ interface, and strong adsorptive capability. This electrochemical biosensor exhibits a detection limit of 0.024 μM, in a linear range of 0.1-382.72 μM with a reproducibility of 2.45 % relative standard deviation, and a good recovery from 96.69 % to 105.59 %, the acquired results ensure a better performance compared with previous reports. The enriched electrocatalytic activity of this developed biosensor is mainly credited to its high surface area, better electrical conductivity, synergistic effect, and abundant electroactive sites. The real-world utility of the VC@Ru-PANI-NPs/SPE-based sensor was ensured via the investigation of biomonitoring of acetaminophen in human blood samples with satisfactory recoveries., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

This study investigates the behavior and intracellular changes in Escherichia coli (model organism) during electro-oxidation with Ti/Sb-SnO 2 /PbO 2 anode in a chlorine free electrochemical system. Preliminary studies were conducted to understand the effect of initial E. coli concentration and applied current density on disinfection. At an applied current density 30 mA cm -2 , 7 log reduction of E. coli was achieved in 75 min. The role of reactive oxygen species' (ROS) in E.coli disinfection was evaluated, which confirmed hydroxyl (•OH) radical as the predominant ROS in electro-oxidation. Observations were carried out at cell and molecular level to understand E.coli inactivation mechanism. Scanning electron microscopy images confirmed oxidative damage of the cell wall and irreversible cell death. Intracellular and extracellular protein quantification and genetic material release further confirmed cell component leakage due to cell wall rupture and degradation due to •OH radical interaction. Change in cell membrane potential suggests the colloidal nature of E. coli cells under applied current density. Plasmid deoxyribonucleic acid degradation study confirmed fragmentation and degradation of released genetic material. Overall, effective disinfection could be achieved by electro-oxidation, which ensures effective inactivation and prevents regrowth of E. coli. Disinfection of real wastewater was achieved in 12 min at an applied current density 30 mA cm -2 . Real wastewater study further confirmed that effective disinfection is possible with a low cost electrode material such as Ti/Sb-SnO 2 /PbO 2 . Energy consumed during disinfection was determined to be 4.978 kWh m -3 for real wastewater disinfection at applied current density 30 mA cm -2 . Cost of operation was estimated and stability of the electrode was studied to evaluate the feasibility of large scale operation. Relatively low energy and less disinfection time makes this technology suitable for field scale applications., (Copyright © 2022 Elsevier Inc. All rights reserved.)

The ICH M7 (R1) guideline recommends the use of complementary (Q)SAR models to assess the mutagenic potential of drug impurities as a state-of-the-art, high-throughput alternative to empirical testing. Additionally, it includes a provision for the application of expert knowledge to increase prediction confidence and resolve conflicting calls. Expert knowledge, which considers structural analogs and mechanisms of activity, has been valuable when models return an indeterminate (equivocal) result or no prediction (out-of-domain). A retrospective analysis of 1002 impurities evaluated in drug regulatory applications between April 2017 and March 2019 assessed the impact of expert review on (Q)SAR predictions. Expert knowledge overturned the default predictions for 26% of the impurities and resolved 91% of equivocal predictions and 75% of out-of-domain calls. Of the 261 overturned default predictions, 15% were upgraded to equivocal or positive and 79% were downgraded to equivocal or negative. Chemical classes with the most overturns were primary aromatic amines (46%), aldehydes (45%), Michael-reactive acceptors (37%), and non-primary alkyl halides (33%). Additionally, low confidence predictions were the most often overturned. Collectively, the results suggest that expert knowledge continues to play an important role in an ICH M7 (Q)SAR prediction workflow and triaging predictions based on chemical class and probability can improve (Q)SAR review efficiency., (Copyright © 2021. Published by Elsevier Inc.)

A dynamic model to predict the germination and outgrowth of Clostridium botulinum spores in cooked ground beef was presented. Raw ground beef was inoculated with a ten-strain C. botulinum spore cocktail to achieve approximately 2 log spores/g. The inoculated ground beef was vacuum packaged, cooked to 71 °C to heat shock the spores, cooled to below 10 °C, and incubated isothermally at temperatures from 10 to 46 °C. C. botulinum growth was quantified and fitted into the primary Baranyi Model. Secondary models were fitted to maximum specific growth rate and lag phase duration using Modified Ratkowsky equation (R 2 0.96) and hyperbolic function (R 2 0.94), respectively. Similar experiments were also performed under non-isothermal (cooling) conditions. Acceptable zone prediction (APZ) analysis was conducted on growth data collected over 3 linear cooling regimes from the current study. The model performance (prediction errors) for all 22 validation data points collected in the current work were within the APZ limits (-1.0 to +0.5 log CFU/g). Additionally, two other growth data sets of C. botulinum reported in the literature were also subjected to the APZ analysis. In these validations, 20/22 and 10/14 predictions fell within the APZ limits. The model presented in this work can be employed to predict C. botulinum spore germination and growth in cooked uncured beef under non-isothermal conditions. The beef industry processors and food service organizations can utilize this predictive microbial model for cooling deviations and temperature abused situations and in developing customized process schedules for cooked, uncured beef products., (Copyright © 2020. Published by Elsevier Ltd.)

Healing or reconstruction of critical-sized bone defects is still challenging in orthopaedic practice. In this study, we developed a new approach to control the degradation and improve the bone regeneration of the AZ31 magnesium substrate, fabricated as mesh cage implants. Subsequently, bilayer nanocomposite coating was carried out using polycaprolactone (PCL) and nano-hydroxyapatite (nHA) by dip-coating and electrospinning. Lastly, the healing capacity of the implants was studied in New Zealand White (NZW) rabbit critical-sized femur bone defects. X-ray analysis showed the coated implant group bridged and healed the critical defects 100% during four weeks of post-implantation. Micro-computed tomography (Micro-CT) study showed higher total bone volume (21.10%), trabecular thickness (0.73), and total porosity (85.71%) with bilayer coated implants than uncoated. Our results showed that nanocomposite coated implants controlled the in vivo degradation and improved bioactivity. Hence, the coated implants can be used as a promising bioresorbable implant for critical segmental bone defect repair applications., (Copyright © 2020 Elsevier Inc. All rights reserved.)

This study describes the preparation of nano-magnesium phosphate (nMP) flakes by one step microwave irradiation method. The synthesized nMP was incorporated with polycaprolactone (PCL), hyperbranched polyglycerol (HPG) and nano-hydroxyapatite (nHA) to fabricate as composite electrospun nanofibrous scaffold for bone tissue engineering applications. The electrospun nanofibers were analyzed by scanning electron microscope, XRD, FTIR, DSC, TGA, and wettability measurement. The nanofibers were smooth, randomly oriented, and surface decorated with nMP. The water contact angle was 32 ± 1° (initial contact angle), which reduces to 0° after 1 min for HPG and nMP containing nanocomposites, while the contact angle of PCL is 104 ± 5°. The nanocomposite scaffolds exhibited higher swelling, biomineralization, and breakages during PBS immersion. The scaffolds were non-cytotoxic to MG63 osteosarcoma cells and hMSCs with higher viability after 72 h. They allowed good adhesion and spreading of these cells when compared to PCL and PCL/nHA electrospun nanofibers. These results indicated that HPG with surface decorated nMP electrospun nanocomposite scaffold can be a promising material for bone tissue engineering applications., Competing Interests: Declaration of competing interest The authors declare there are no conflicts of interest regarding the publication of this paper., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Post-harvest washing of produce is performed to remove physical debris and to lower microbial load. The use of ozone in combination with plant-based antimicrobials was evaluated as an alternative to conventional sanitizers such as chlorine. Plant based antimicrobials that were evaluated in combination with ozone included oregano oil, carvacrol, Quillaja saponin and olive extract. Ozone was dispersed in phosphate buffered saline (PBS), following which individual antimicrobials or their combinations were added. Iceberg lettuce leaves (10 g portions) inoculated with Salmonella enterica serotype Newport (6.5 ± 1 log CFU/g) were added to the wash suspension. The leaves were tested for reduction in S. Newport population after 60, 90 and 120 min of treatment. Exposure to ozonized water for 120 min resulted in a 2.1 log CFU/g (p < 0.05) reduction in S. Newport population. The addition of 0.1% oregano oil to ozonized water resulted in 3 log CFU/g reduction after 120 min but a 4.1 log CFU/g reduction after 60 min, indicating that the antioxidant property of oregano oil might have diminished ozone activity and resuscitated injured S. Newport cells. The addition of 5% olive extract to ozonized water resulted in 4.2 log CFU/g reduction of S. Newport after 120 min (p < 0.05) of treatment. While 5% olive extract did not confer protection to S. Newport cells from ozone, 1% olive extract resulted in higher S. Newport survival after 120 min treatment than the 60 min treatment. The use of carvacrol (0.1%, 0.3% and 0.5%) in ozonized water reduced the pathogen population to below the limit of detection (10 CFU/g) (p < 0.05) which was in excess of 6 log CFU/g. These results indicate that the efficacy of ozone is compounded by the addition of certain plant-based antimicrobials when used at optimum concentrations. Ozone combined with plant antimicrobials could serve as an effective alternative to sanitizers currently used for washing and processing of produce., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The major modifiable risk factor in melanomagenesis is UV exposure and mutagenesis of melanocytes. Other UV-induced events that contribute to early tumorigenesis are poorly understood. Herein we show that the repeated exposure of human primary melanocytes to UVB results in a sustained senescence response, increases in expression of signal transducer and activator of transcription 1, MX1, OAS2, and IRF7 proteins of up to 75-fold, and resistance to subsequent UVB-induced apoptosis. In the setting of UVB-induced DNA damage, we detected time-dependent increases in the release of damage-associated molecular patterns such as high-mobility group box 1 (HMGB1). After intermittent UVB exposure, melanocytes treated with the JAK inhibitor ruxolitinib reduced expression of HMGB1 and MX1 as well as activation of JAK1 (pJAK1) and signal transducer and activator of transcription 1 (pSTAT1). In addition, melanocytes expressing small hairpin RNA selective for the HMGB1 receptor, receptor for advanced glycosylation end product (RAGE), exhibited decreased expression of both HMGB1 and MX1 after UVB exposure. The response of small hairpin RAGE-infected cells to human recombinant HMGB1 was blunted with decreased MX1 expression and JAK activation. Finally, depletion of receptor for advanced glycosylation end product decreased UVB-induced resistance to apoptosis (P < 0.05). These findings highlight a cell autonomous response to UV damage, contribute to their resistance to apoptosis and cell death, and may have implications for early stages of melanoma development., (Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.)

This current study is aimed towards the fabrication of AZ31 magnesium cylindrical mesh cage implant with circular holes for orthopedic applications. This mesh cage is coated with nanocomposite material containing polycaprolactone (PCL), pluronic F127 and nano hydroxyapatite (nHA) by electrospinning process. Morphology and composition were analyzed by various characterization techniques. Controlled degradation and weight loss of the nanocomposite coated samples in 28 days were observed when compared with uncoated samples in SBF (simulated body fluid). The nanocomposite coated material was not cytotoxic to MG63 osteosarcoma cells. The cell viability, morphology, ALP activity, calcium mineralization and collagen deposition were also better on this when compared to uncoated. Smooth and randomly deposited nanofibers on the mesh cage was observed and the contact angle indicated that the surface is hydrophilic with (initial contact angle of 55 ± 1° and after 10 s 0°) when compared to PCL (99°) coated surface. 2-5 fold higher mRNA expression levels of osteogenic genes namely ALP, BMP2, COL1 and RUNX2 was observed with nanocomposite coated scaffolds than uncoated and PCL coated samples in 14 days. These results indicate the potential use of the nanocomposite coated AZ31 cylindrical mesh cage for segmental bone defect repair and can be used as a degradable implant for orthopedic applications., (Copyright © 2018. Published by Elsevier B.V.)

A novel one-step preparation of magnesium particles and Pluronic F127 incorporated with calcium sulfate hemihydrate (CSH) and nano-hydroxyapatite (nHA) ready to use injectable or moldable beads was developed for bone tissue regeneration applications. The nanocomposite showed setting time less than 15 min, very good injectability (75-85%) and good mechanical strength (52-80 MPa). Samples immersed in SBF showed controlled degradation (40-45% reduction in weight) in 28 days. The nanocomposite bone graft was cytocompatible against MG63 osteosarcoma cells and increased the osteogenic gene expression by 2-3 folds. These results indicate that it can be a potential defect filling biomaterial for bone tissue regeneration at the fracture site., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

This study is aimed to develop curcumin (Cur) incorporated electrospun nanofibers of a blend of poly (lactic acid) (PLA) and hyperbranched polyglycerol (HPG) for wound healing applications. Both the polymers are synthesized and fabricated by electrospinning technique. The produced nanofibers were characterized by Scanning Electron Microscopy (SEM), X-Ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FT-IR), Differential Scanning Colorimetry (DSC) and Thermogravimetric Analysis (TGA). Electrospun scaffolds (PLA/HPG/Cur) exhibits very high hydrophilicity, high swelling and drug uptake and promotes better cell viability, adhesion and proliferation when compared to PLA/Cur electrospun nanofibers. Biodegradation study revealed that the morphology of the nanofibers were unaffected even after 14days immersion in Phosphate Buffered Saline. In vitro scratch assay indicates that migration of the cells in the scratch treated with PLA/HPG/Cur is complete within 36h. These results suggest that PLA/HPG/Cur nanofibers can be a potential wound patch dressing for acute and chronic wound applications., (Copyright © 2017 Elsevier B.V. All rights reserved.)

AZ31 magnesium alloy was coated with polycaprolactone (PCL) nano-fibrous layer using electrospinning technique so as to control degradation in physiological environment. Before coating, the alloy was treated with HNO3 to have good adhesion between the coating and substrate. To elucidate the role of pre-treatment and coating, samples only with PCL coating as well as HNO3 treatment only were prepared for comparison. Best coating adhesion of 4B grade by ASTM D3359-09 tape test was observed for pre-treated samples. The effect of coating on in vitro degradation and biomineralization was studied using supersaturated simulated body fluid (SBF 5×). The weight loss and corrosion results obtained by immersion test showed that the combination of HNO3 pre-treatment and PCL coating is very effective in controlling the degradation rate and improving bioactivity. Cytotoxicity studies using L6 cells showed that PCL coated sample has better cell adhesion and proliferation compared to uncoated samples. Nano-fibrous PCL coating combined with prior acid treatment seems to be a promising method to tailor degradation rate with enhanced bioactivity of Mg alloys., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Water samples from three rivers in southern India were analyzed for octylphenol (OP), nonylphenol (NP), and bisphenol A (BPA) residues by gas chromatography and mass spectrometry. The concentrations of OP, NP and BPA ranged from ND (not detected) to 16.3 ng/L, ND to 2200 ng/L, and 2.8 to 136 ng/L with detection frequencies of 96%, 66% and 100%, respectively. All three rivers showed a similar distribution pattern of NP>>BPA>OP, however, the Kaveri river had elevated levels. The phenolic compounds in rivers are expected to cause potential toxicity to aquatic organism including crustaceans, molluscs, insects and fish. In respect to aquatic risk assessment, NP showed a greater hazard quotient (HQ) than did OP and BPA, and the highest HQ (62) was observed for fish in the Kaveri river. The backwater and estuarine NP levels may pose a risk to larvae of oysters and barnacles. Based on tolerable daily intake and reference dose, BPA and NP levels are considered safe for humans. However, NP levels in some water samples were greater than the drinking water safety limit (0.5 µg/L). This is the first report on phenolic compounds and their associated aquatic risks in Indian rivers., (© 2013 Published by Elsevier Inc.)

Lung carcinogenesis in humans involves an accumulation of genetic and epigenetic changes that lead to alterations in normal lung epithelium, to in situ carcinoma, and finally to invasive and metastatic cancers. The loss of transforming growth factor β (TGF-β)-induced tumor suppressor function in tumors plays a pivotal role in this process, and our previous studies have shown that resistance to TGF-β in lung cancers occurs mostly through the loss of TGF-β type II receptor expression (TβRII). However, little is known about the mechanism of down-regulation of TβRII and how histone deacetylase (HDAC) inhibitors (HDIs) can restore TGF-β-induced tumor suppressor function. Here we show that HDIs restore TβRII expression and that DNA hypermethylation has no effect on TβRII promoter activity in lung cancer cell lines. TGF-β-induced tumor suppressor function is restored by HDIs in lung cancer cell lines that lack TβRII expression. Activation of mitogen-activated protein kinase/extracellular signal-regulated kinase pathway by either activated Ras or epidermal growth factor signaling is involved in the down-regulation of TβRII through histone deacetylation. We have immunoprecipitated the protein complexes by biotinylated oligonucleotides corresponding to the HDI-responsive element in the TβRII promoter (-127/-75) and identified the proteins/factors using proteomics studies. The transcriptional repressor Meis1/2 is involved in repressing the TβRII promoter activity, possibly through its recruitment by Sp1 and NF-YA to the promoter. These results suggest a mechanism for the downregulation of TβRII in lung cancer and that TGF-β tumor suppressor functions may be restored by HDIs in lung cancer patients with the loss of TβRII expression.

Pharmaceutical and personal care products are used extensively worldwide and their residues are frequently reported in aquatic environments. In this study, antiepileptic, antimicrobial and preservative compounds were analyzed in surface water and sediment from the Kaveri, Vellar and Tamiraparani rivers, and in the Pichavaram mangrove in India by gas chromatography-mass spectrometry (GC-MS). The mean concentration of carbamazepine recorded in the Kaveri River water (28.3 ng/L) was higher than in the other rivers and the mangrove. Because carbamazepine is used only in human drugs, this may reflect the relative contributions of human excretions/sewage in these rivers. The mean triclosan level in the Tamiraparani River (944 ng/L) was an order of magnitude greater than in the other water systems, and the concentrations at two of the sites reported here (3800-5160 ng/L) are, to our best knowledge, among the highest detected in surface waters. Sediment levels were, however, comparable with other sites. We conclude that industrial releases are likely major contributors of triclosan into this river system. Among parabens, ethyl paraben was predominantly observed. Hazard Quotients suggest greater environmental risks for triclosan than for carbamazepine and parabens. This is the first study on antiepileptic, antimicrobial and preservatives in rivers and mangroves from India., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Background: Prostate cancer is one of the most frequently diagnosed cancers in men. Progression of these tumors is facilitated by growth factors that activate critical signaling cascades thereby promote prostate cancer cell growth, survival, and migration. Among these, insulin-like growth factors (IGFs) signaling pathway contributes a major role. In this study, we examined the effect of zinc on insulin-like growth factors signaling in prostate cancer cells., Methods: Human androgen-independent prostatic carcinoma (PC-3) cells were treated with different concentrations of zinc (20-100micromol/l) for 24 and 48h. Cell viability was performed by 3[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Insulin-like growth factor binding protein-3 (IGFBP-3), insulin-like growth factor-I receptor (IGF-IR), insulin receptor substrate-1 (IRS-1) and IRS-2, phosphatidylinositol-3 kinase (PI-3 K), protein kinase B or Akt, phosphorylated Akt (p-Akt), extracellular regulated kinase 1/2 (ERK1/2), phosphorylated ERK1/2 (p-ERK1/2), and cyclin D1 protein levels were assessed by Western blot analysis. Apoptosis was confirmed by 4',6'-diaminido-2-phenylindole dihydrochloride (DAPI) staining, and mitochondrial membrane potential was performed using rhodamine-123 staining method., Results: Zinc significantly reduces the cell viability of PC-3 cells. It decreases the protein levels of IGF-IR, IRS-1, and IRS-2 and increases the level of IGFBP-3. Zinc reduces the levels of PI-3 K, Akt, ERK1/2, and cyclin Dl. Loss of mitochondrial membrane potential and apoptotic cell death were also observed in zinc-treated cells., Conclusion: This study suggests that zinc decreases the survival of androgen-independent prostate cancer cells by modulating the expression of IGF system components and its signaling molecules. Thus, zinc may be qualified as a potential agent for the treatment of prostate cancer., (Copyright 2009 Elsevier B.V. All rights reserved.)

A role for oestrogen in regulating fluid reabsorption in the monkey epididymis was recently demonstrated. Here, these studies are extended to identify potential oestrogen-regulated proteins in the cauda region of monkey epididymis treated with vehicle and oestrogen receptor antagonist (ICI 182780). Two-dimensional electrophoretic analysis was used to identify the proteins. The results indicated down-regulation of WNT4 in the ICI-182780-treated monkey cauda. In addition, the Wnt4 mRNA concentration was also reduced in the caput regions of ICI-182780- treated rats and oestrogen receptor knockout mice. WNT4 is a key regulator of gonadal differentiation in humans and mice and plays a pivotal role in early mouse embryogenesis. The results of the present study establish the presence of WNT4 in the monkey epididymis and its regulation by oestrogen, and suggest a role for WNT4 in maintaining epididymal homeostasis.