Clearing-enabled light sheet microscopy as a novel method for three-dimensional mapping of the sensory innervation of the mouse knee.

A major barrier that hampers our understanding of the precise anatomic distribution of pain sensing nerves in and around the joint is the limited view obtained from traditional two dimensional (D) histological approaches. Therefore, our objective was to develop a workflow that allows examination of the innervation of the intact mouse knee joint in 3D by employing clearing-enabled light sheet microscopy. We first surveyed existing clearing protocols (SUMIC, PEGASOS, and DISCO) to determine their ability to clear the whole mouse knee joint, and discovered that a DISCO protocol provided the most optimal transparency for light sheet microscopy imaging. We then modified the DISCO protocol to enhance binding and penetration of antibodies used for labeling nerves. Using the pan-neuronal PGP9.5 antibody, our protocol allowed 3D visualization of innervation in and around the mouse knee joint. We then implemented the workflow in mice intra-articularly injected with nerve growth factor (NGF) to determine whether changes in the nerve density can be observed. Both 3D and 2D analytical approaches of the light sheet microscopy images demonstrated quantifiable changes in midjoint nerve density following 4 weeks of NGF injection in the medial but not in the lateral joint compartment. We provide, for the first time, a comprehensive workflow that allows detailed and quantifiable examination of mouse knee joint innervation in 3D.

Cells transiently expressing periostin are required for intramedullary intramembranous bone regeneration.

Intramembranous bone regeneration plays an important role in fixation of intramedullary implants used in joint replacement and dental implants used in tooth replacement. Despite widespread recognition of the importance of intramembranous bone regeneration in these clinical procedures, the underlying mechanisms have not been well explored. A previous study that examined transcriptomic profiles of regenerating bone from the marrow space showed that increased periostin gene expression preceded increases in several osteogenic genes. We therefore sought to determine the role of cells transiently expressing periostin in intramedullary intramembranous bone regeneration. We used a genetic mouse model that allows tamoxifen-inducible fluorescent labeling of periostin expressing cells. These mice underwent ablation of the bone marrow cavity through surgical disruption, a well-established intramembranous bone regeneration model. We found that in intact bones, fluorescently labeled cells were largely restricted to the periosteal surface of cortical bone and were absent in bone marrow. However, following surgical disruption of the bone marrow cavity, cells transiently expressing periostin were found within the regenerating tissue of the bone marrow compartment even though the cortical bone remained intact. The source of these cells is likely heterogenous, including cells occupying the periosteal surface as well as pericytes and endothelial cells within the marrow cavity. We also found that diphtheria toxin-mediated depletion of cells transiently expressing periostin at the time of surgery impaired intramembranous bone regeneration in mice. These data suggest a critical role of periostin expressing cells in intramedullary intramembranous bone regeneration and may lead to novel therapeutic interventions to accelerate or enhance implant fixation.

Mycotoxins in food and feed: toxicity, preventive challenges, and advanced detection techniques for associated diseases.

Mycotoxins are produced primarily as secondary fungal metabolites. Mycotoxins are toxic in nature and naturally produced by various species of fungi, which usually contaminate food and feed ingredients. The growth of these harmful fungi depends on several environmental factors, such as pH, humidity, and temperature; therefore, the mycotoxin distribution also varies among global geographical areas. Various rules and regulations regarding mycotoxins are imposed by the government bodies of each country, which are responsible for addressing global food and health security concerns. Despite this legislation, the incidence of mycotoxin contamination is continuously increasing. In this review, we discuss the geographical regulatory guidelines and recommendations that are implemented around the world to control mycotoxin contamination of food and feed products. Researchers and inventors from various parts of the world have reported several innovations for controlling mycotoxin-associated health consequences. Unfortunately, most of these techniques are restricted to laboratory scales and cannot reach users. Consequently, to date, no single device has been commercialized that can detect all mycotoxins that are naturally available in the environment. Therefore, in this study, we describe severe health hazards that are associated with mycotoxin exposure, their molecular signaling pathways and processes of toxicity, and their genotoxic and cytotoxic effects toward humans and animals. We also discuss recent developments in the construction of a sensitive and specific device that effectively implements mycotoxin identification and detection methods. In addition, our study comprehensively examines the recent advancements in the field for mitigating the health consequences and links them with the molecular and signaling pathways that are activated upon mycotoxin exposure.

Colon epithelial cell-specific Bmal1 deletion impairs bone formation in mice.

The circadian clock system regulates multiple metabolic processes, including bone metabolism. Previous studies have demonstrated that both central and peripheral circadian signaling regulate skeletal growth and homeostasis in mice. Disruption in central circadian rhythms has been associated with a decline in bone mineral density in humans and the global and osteoblast-specific disruption of clock genes in bone tissue leads to lower bone mass in mice. Gut physiology is highly sensitive to circadian disruption. Since the gut is also known to affect bone remodeling, we sought to test the hypothesis that circadian signaling disruption in colon epithelial cells affects bone. We therefore assessed structural, functional, and cellular properties of bone in 8 week old Ts4-Cre and Ts4-Cre;Bmal1fl/fl (cBmalKO) mice, where the clock gene Bmal1 is deleted in colon epithelial cells. Axial and appendicular trabecular bone volume was significantly lower in cBmalKO compared to Ts4-Cre 8-week old mice in a sex-dependent fashion, with male but not female mice showing the phenotype. Similarly, the whole bone mechanical properties were deteriorated in cBmalKO male mice. The tissue level mechanisms involved suppressed bone formation with normal resorption, as evidenced by serum markers and dynamic histomorphometry. Our studies demonstrate that colon epithelial cell-specific deletion of Bmal1 leads to failure to acquire trabecular and cortical bone in male mice.

Photodynamic treatment of multidrug-resistant bacterial infection using indium phosphide quantum dots.

Infections caused by multidrug-resistant (MDR) bacteria pose an impending threat to humanity, as the evolution of MDR bacteria outpaces the development of effective antibiotics. In this work, we use indium phosphide (InP) quantum dots (QDs) to treat infections caused by MDR bacteria via photodynamic therapy (PDT), which shows superior bactericidal efficiency over common antibiotics. PDT in the presence of InP QDs results in high-efficiency bactericidal activity towards various bacterial species, including Staphylococcus aureus, Bacillus cereus, Escherichia coli and Pseudomonas aeruginosa. Upon light absorption, InP QDs generate superoxide (O2˙-), which leads to efficient and selective killing of MDR bacteria while mammalian cells remain intact. The cytotoxicity evaluation reveals that InP QDs are bio- and blood-compatible in a wide therapeutic window. For the in vivo study, we drop a solution of InP QDs at a concentration within the therapeutic window onto MDR S. aureus-infected skin wounds of mice and perform PDT for 15 min. InP QDs show excellent therapeutic and prophylactic efficacy in treating MDR bacterial infection. These findings show that InP QDs have great potential to serve as antibacterial agents for MDR bacterial infection treatment, as an effective and complementary alternative to conventional antibiotics.

Evolution of Highly Biocompatible and Thermally Stable YVO4:Er3+/Yb3+ Upconversion Mesoporous Hollow Nanospheriods as Drug Carriers for Therapeutic Applications.

In recent times, upconversion nanomaterials with mesoporous hollow structures have gained significant interest as a prospective nano-platform for cancer imaging and therapeutic applications. In this study, we report a highly biocompatible YVO4:1Er3+/10Yb3+ upconversion mesoporous hollow nanospheriods (YVO4:Er3+/Yb3+ UC-MHNSPs) by a facile and rapid self-sacrificing template method. The Rietveld analysis confirmed their pure phase of tetragonal zircon structure. Nitrogen adsorption-desorption isotherms revealed the mesoporous nature of these UC-MHNSPs and the surface area is found to be ~87.46 m2/g. Under near-infrared excitation (980 nm), YVO4:Er3+/Yb3+ UC-MHNSPs showed interesting color tunability from red to green emission. Initially (at 0.4 W), energy back transfer from Er3+ to Yb3+ ions leads to the strong red emission. Whereas at high pump powers (1 W), a fine green emission is observed due to the dominant three-photon excitation process and traditional energy transfer route from Er3+ to Yb3+ ions. The bright red light from the membrane of HeLa cells confirmed the effective cellular uptake of YVO4:Er3+/Yb3+ UC-MHNSPs. The resonant decrease in cell viability on increasing the concentration of curcumin conjugated YVO4:Er3+/Yb3+ UC-MHNSPs established their excellent antitumor activity. Therefore, the acquired results indicate that these YVO4:Er3+/Yb3+ UC-MHNSPs are promising drug carriers for bioimaging and various therapeutic applications.

An Overview on Single-Cell Technology for Hepatocellular Carcinoma Diagnosis.

Hepatocellular carcinoma is a primary liver cancer caused by the accumulation of genetic mutation patterns associated with epidemiological conditions. This lethal malignancy exhibits tumor heterogeneity, which is considered as one of the main reasons for drug resistance development and failure of clinical trials. Recently, single-cell technology (SCT), a new advanced sequencing technique that analyzes every single cell in a tumor tissue specimen, aids complete insight into the genetic heterogeneity of cancer. This helps in identifying and assessing rare cell populations by analyzing the difference in gene expression pattern between individual cells of single biopsy tissue which normally cannot be identified from pooled cell gene expression pattern (traditional sequencing technique). Thus, SCT improves the clinical diagnosis, treatment, and prognosis of hepatocellular carcinoma as the limitations of other techniques impede this cancer research progression. Application of SCT at the genomic, transcriptomic, and epigenomic levels to promote individualized hepatocellular carcinoma diagnosis and therapy. The current review has been divided into ten sections. Herein we deliberated on the SCT, hepatocellular carcinoma diagnosis, tumor microenvironment analysis, single-cell genomic sequencing, single-cell transcriptomics, single-cell omics sequencing for biomarker development, identification of hepatocellular carcinoma origination and evolution, limitations, challenges, conclusions, and future perspectives.

Partners in crime: The Lewis Y antigen and fucosyltransferase IV in Helicobacter pylori-induced gastric cancer.

Helicobacter pylori (H. pylori) is a major causative agent of chronic gastritis, gastric ulcer and gastric carcinoma. H. pylori cytotoxin associated antigen A (CagA) plays a crucial role in the development of gastric cancer. Gastric cancer is associated with glycosylation alterations in glycoproteins and glycolipids on the cell surface. H. pylori cytotoxin associated antigen A (CagA) plays a significant role in the progression of gastric cancer through post-translation modification of fucosylation to develop gastric cancer. The involvement of a variety of sugar antigens in the progression and development of gastric cancer has been investigated, including type II blood group antigens. Lewis Y (LeY) is overexpressed on the tumor cell surface either as a glycoprotein or glycolipid. LeY is a difucosylated oligosaccharide, which is catalyzed by fucosyltransferases such as FUT4 (α1,3). FUT4/LeY overexpression may serve as potential correlative biomarkers for the prognosis of gastric cancer. We discuss the various aspects of H. pylori in relation to fucosyltransferases (FUT1-FUT9) and its fucosylated Lewis antigens (LeY, LeX, LeA, and LeB) and gastric cancer. In this review, we summarize the carcinogenic effect of H. pylori CagA in association with LeY and its synthesis enzyme FUT4 in the development of gastric cancer as well as discuss its importance in the prognosis and its inhibition by combination therapy of anti-LeY antibody and celecoxib through MAPK signaling pathway preventing gastric carcinogenesis.

Cellular antioxidant potential and inhibition of foodborne pathogens by a sesquiterpene ilimaquinone in cold storaged ground chicken and under temperature-abuse condition.

A sesquiterpene quinone, ilimaquinone, was accessed for its cellular antioxidant efficacy and possible antimicrobial mechanism of action against foodborne pathogens (Staphylococcus aureus and Escherichia coli) in vitro and in vivo. Ilimaquinone was found to be protective against H2O2-induced oxidative stress as validated by the reduction in the ROS levels, including increasing expression of SOD1 and SOD2 enzymes. Furthermore, ilimaquinone evoked MIC against S. aureus and E. coli within the range of 125-250 µg/mL. Ilimaquinone established its antimicrobial mode of action against both tested pathogens as evident by bacterial membrane depolarization, loss of nuclear genetic material, potassium ion, and release of extracellular ATP, as well as compromised membrane permeabilization and cellular component damage. Also, ilimaquinone showed no teratogenic effect against zebrafish, suggesting its nontoxic nature. Moreover, ilimaquinone significantly reduced the S. aureus count without affecting the sensory properties and color values of cold-storaged ground chicken meat even under temperature abuse condition.

The anti-cancerous activity of adaptogenic herb Astragalus membranaceus.

BACKGROUND: Cancer is the most dreadful disease increasing rapidly causing an economic burden globally. A standardized chemotherapy regimen planned with curative intent weakens the immune system and damages healthy cells making the patient prone to infections and severe side effects with pain and fatigue. PURPOSE: Astragalus membranaceus (AM) has a long history of use in the treatment of severe adverse diseases. For thousands of years, it has been used in mixed herbal decoctions for the treatment of cancer. Due to growing interest in this plant root for its application to treat various types of cancers and tumors, has attracted researcher's interest. METHOD: The literature search was done from core collections of electronic databases such as Web of Science, Google Scholar, PubMed and Science Direct using keywords given below and terms like pharmacological and phytochemical details of this plant. OUTCOME: Astragalus membranaceus has demonstrated the ability to modulate the immune system during drug therapy making the patient physically fit and prolonged life. It has become a buzzword of herbalists as it is one of the best of seven important adaptogenic herbs with a protective effect against chronic stress and cancer. It demonstrated significant amelioration of the perilous toxic effects induced by concurrently administered chemo onco-drugs. CONCLUSION: The natural phytoconstituents of this plant formononetin, astragalus polysaccharide, and astragalosides which show high potential anti-cancerous activity are studied and discussed in detail. One of them are used in clinical trials to overcome cancer related fatigue. Overall, this review aims to provide an insight into Astragalus membranaceus status in cancer therapy.

Targeting key transcriptional factor STAT3 in colorectal cancer.

In developed countries, colorectal cancer (CRC) is the fourth most common cancer and the second leading cause of malignant-related deaths. CRC is treatable cancer when diagnosed early; however, diagnosis at the advanced stage is associated with a poor prognosis. Although chemotherapy is generally very promising, STAT3 protein which is overexpressed and persistently activated in CRC cells is observed to be the major contributor of chemoresistance development. It has been shown to play a prominent and pathogenic role in CRC initiation, progression, and metastasis. While over the past few years, research has been focused on STAT3 which is expressed at the center of various oncogenic pathways. This review is a discussion of the oncogenic role of STAT3 in CRC and potential therapeutic STAT3 inhibitors and analogs used to control and treat CRC.

Reproductive toxic potential of phthalate compounds - State of art review.

Phthalates are pervasive compounds, and due to the ubiquitous usage of phthalates, humans or even children are widely exposed to them. Since phthalates are not chemically bound to the plastic matrix, they can easily leach out to contaminate the peripheral environment. Various animal and human studies have raised vital health concern including developmental and reproductive toxicity of phthalate exposure. The present review is based upon the available literature on phthalates with respect to their reproductive toxic potential. Common reproductive effects such as declined fertility, reduced testis weight, variations in accessory sex organs and several female reproductive disorders appeared to be largely associated with the transitional phthalates. Among the higher molecular weight phthalates (≥ C7), di-isononyl phthalate (DINP) produces some minor effects on development of male reproductive tract and among low molecular weight phthalates (≤C3), di-methyl (DMP) and di-isobutyl (DIBP) phthalate produce some adverse effects on male reproductive system. Whereas transitional phthalates such as di-butyl phthalate, benzyl butyl phthalate, and di-(2-ethylhexyl) phthalate have shown adverse effects on female reproductive system. Owing to these, non-toxic alternatives to phthalates may be developed and use of phthalates could be rationalized as an important issue where human reproduction system is involved. Though, more epidemiological studies are needed to substantiate the reported findings on phthalates.

Metasequoia glyptostroboides potentiates anticancer effect against cervical cancer via intrinsic apoptosis pathway.

This study was undertaken to investigate the anticancer effects of organic extracts derived from the floral cones of Metasequoia glyptostroboides. Dried powder of M. glyptostroboides floral cones was subjected to methanol extraction, and the resulting extract was further partitioned by liquid-liquid extraction using the organic solvents n-hexane, dichloromethane (DME), chloroform, and ethyl acetate in addition to deionized water. HeLa cervical and COS-7 cells were used as a cancer cell model and normal cell control, respectively. The anticancer effect was evaluated by using the Cell Counting Kit-8 assay. The viability of COS-7 cells was found to be 12-fold higher than that of the HeLa cells under the administration of 50 µg/ml of the DME extract. Further, the sub-G1 population was determined by FACS analysis. The number of cells at the sub-G1 phase, which indicates apoptotic cells, was increased approximately fourfold upon treatment with the DME and CE extracts compared with that in the negative control. Furthermore, RT-qPCR and western blotting were used to quantitate the relative RNA and protein levels of the cell death pathway components, respectively. Our results suggest that the extracts of M. glyptostroboides floral cones, especially the DME extract, which possesses several anticancer components, as determined by GC-MS analysis, could a potential natural anticancer agent.

Antioxidant and anti-aging potential of a peptide formulation (Gal2-Pep) conjugated with gallic acid.

Skin is highly vulnerable to premature aging due to external stress, therefore, in this study, a peptide formulation, (galloyl)2-KTPPTTP (Gal2-Pep) was synthesized by combining TPPTTP peptide, and gallic acid (GA). All peptides were synthesized on 2-chlorotrityl chloride resin using solid-phase peptide synthesis (SPPS), and analyzed on an electrospray ionization (ESI)/quadrupole-time-of-flight (Q-TOF) tandem mass spectroscopy (MS) system. Initially, Gal2-Pep showed no toxicity below the concentration 100 µM with cell survival rate of 88% for keratinocytes and fibroblasts. The reactive oxygen species (ROS) scavenging activity of Gal2-Pep was more stable compared to GA alone; and after four weeks at room temperature, its ROS scavenging activity remained higher than 50%. Moreover, the peptide formulation, Gal2-Pep also exhibited elastase inhibitory effect in CCD-1064Sk fibroblast cells. Based on the results of RT-qPCR, it was proved in this study that Gal2-Pep increased the expression of PGC-1α to prevent oxidative stress, and validated its potential as an anti-aging agent through increasing the expression of type I collagen and by decreasing the expression of matrix metalloproteinase-1 (MMP1). The findings obtained reinforce the suggestion that the peptide formulation synthesized in this study could be used as a natural antioxidant and anti-aging agent for its cosmetic applications.

Folic acid-modified bovine serum albumin nanoparticles with doxorubicin and chlorin e6 for effective combinational chemo-photodynamic therapy.

We herein describe a facile method to synthesize stable bovine serum albumin-based nanoparticles (BNPs) loaded with two anticancer therapeutics, doxorubicin (DOX) and a photosensitizer, chlorin e6 (Ce6), in combination with folic acid (FA) as a target cancer cell receptor for the development of an effective combined chemo and photodynamic (FA-Ce6/DOX/BNPs) therapy against cervical cancer. FA-Ce6/DOX/BNPs exhibited excellent monodispersity with an average diameter of 103.5 ± 3.8 nm, a negative zeta potential of approximately -30.44 ± 0.35 mV, and long-term stability. As a result, FA-Ce6/DOX/BNPs exhibited severe toxicity to cervical HeLa cancer cells. Also, a higher drug release rate was observed under acidic pH conditions (pH 5.0). Moreover, FA-Ce6/DOX/BNPs potentiated mitochondrial reactive oxygen species (ROS) production in HeLa cells under 671-nm laser exposure, leading to activation of key regulator proteins of apoptosis such as BH3 interacting-domain death agonist (BID), B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X (BAX), as well as induction of the caspase cascade and mitochondrial ROS-mediated cell death. Confocal microscopy analysis further validated cellular uptake of FA-Ce6/DOX/BNPs by HeLa cells. Furthermore, results of real-time quantitative PCR (RT-qPCR) and western blot analysis further validated the anticancer effect of FA-Ce6/DOX/BNPs, as evidenced by elevated gene/protein expression levels of apoptotic biomarkers p53, BID, caspase-3, cleaved poly(ADP-ribose) polymerase 1 (PARP-1), and BAX, contrary to levels of the anti-apoptotic marker Bcl-2. Moreover, in vivo toxicity results of FA-Ce6/DOX/BNPs using laser irradiation in zebrafish larvae, as a chemo-photodynamic therapy confirmed that it does not affect the larval development without causing any adverse toxic effect in zebrafish larvae. Altogether these findings strongly support the anticancer effect of FA-Ce6/DOX/BNPs combinational chemo-photodynamic therapy, which could be a promising candidate for cervical cancer therapy.

N,P-Doped Carbon Nanodots for Food-Matrix Decontamination, Anticancer Potential, and Cellular Bio-Imaging Applications.

We report a facile one-step thermal treatment method for the synthesis of biocompatible, fluorescent nitrogen-phosphorus-doped carbon nanodots (NPCDs) as multifunctional agents for the food matrix decontamination, cancer targeting, and cellular bio-imaging. NPCDs exhibit high toxicity towards L. monocytogenes, as illustrated by fluorescent live-dead cell counting, disruption of membrane permeability/potential, changes in the levels of cellular ions, genetic materials, and proteins, as well as intracellular production of reactive oxygen species. The tryptophan and protein peaks released in NPCDs treated cells contributed to indole ring breathing and correlated with induced cell death. NPCDs significantly inhibited bacterial biofilm formation on a solid substrate. NPCDs-coated low-density polyethylene (LDPE) film crosslinked with 1% aminopropyltriethoxy silane (APTES) via silane-hydroxyl linking as a food-grade wrap significantly reduced bacterial counts in a raw chicken food model. Furthermore, NPCDs induced apoptosis in HeLa cervical cancer cells, as confirmed by the distorted cell morphology, fluorescence microscopic analysis, presence of fragmented nuclei and the qPCR results of mRNA expression levels of apoptotic markers. Moreover, NPCDs were also applicable in utilized for the cellular bio-imaging of KM12-C colon cancer cells under confocal microscopy owing to their excellent luminescence properties. Overall, NPCDs represent a promising platform to reduce the environmental health risks associated with hazardous pathogens, anticancer targeting, and their application in cellular bio-imaging as multifunctional targets/nanocarriers.

Hierarchical N-doped TiO2@Bi2WxMo1-xO6 core-shell nanofibers for boosting visible-light-driven photocatalytic and photoelectrochemical activities.

Heterogeneous photocatalysis has been proven to be a promising approach to overcome the great challenges encountered with conventional technologies for environmental remediation. Herein, for the first time, a novel hierarchical architecture of nitrogen-doped TiO2@Bi2WxMo1-xO6 (N-T@BWMO-x, x = 0-1.0) was rationally designed and fabricated through an electrospinning route followed by a solvothermal process. The photocatalytic activity of the as-prepared samples was evaluated based on the degradation of tetracycline hydrochloride (TC) under visible-light irradiation. The results indicated that the molar fraction of W/Mo has a strong impact on the photocatalytic efficiency and photoelectrochemical performance of the N-T@BWMO composites. Compared to N-TiO2 and the binary composites, N-T@BWMO-0.25 exhibited outstanding photocatalytic activity and significant cycling stability. The enhanced photocatalytic activity can be synergistically linked to the excellent native adsorption, extended light-harvesting region, hierarchical structure, and strong interfacial interaction between N-TiO2 and BWMO, which can effectively prolong the lifetime of charge-carriers. Moreover, active species-trapping and electron paramagnetic resonance results confirmed that holes and superoxide radicals were the dominant active species responsible for TC removal. A possible photocatalytic mechanism underlying the degradation of TC by N-T@BWMO-0.25 is also proposed. We expect that our findings will provide new insights into the use of highly efficient core-shell heterostructure photocatalysts, with potential applications in environmental decontamination.

Fluorescent immunoliposomal nanovesicles for rapid multi-well immuno-biosensing of histamine in fish samples.

Scombroid poisoning in fish-based and other food products has raised concerns due to toxicity outbreaks and incidences associated with histamine, thus measuring the amount of histamine toxic molecule is considered crucial quality indicator of food safety and human health. In this study, liposome-based measurement of histamine was performed via rupturing mechanism of sulforhodamine B dye encapsulated anti-histamine antibody conjugated liposomal nanovesicles. The immunosensing ability of immuno-liposomal format was assessed by monitoring the fluorescence at excitation/emission wavelength of 550/585 nm. Immuno-liposomal format assays were considered, one based on single wash procedure (Method 1), which had a detection limit of 10 ppb and quantification limit 15-80 ppb. While Method 2 based on one-by-one wash procedure had a detection limit of 2-3 ppb and quantification limit 8.5 ppb-200 ppm that required 2 h 30 min to perform. In view of better quantification limit, Method 2 was chosen for further tests required to validate its applicability in real samples. The feasibility of Method 2 was reconfirmed in fresh mackerel fish, and canned fish (tuna and salmon) with a similar detection limits but with low amplified fluorescence signals and sufficient levels of histamine recovery from fresh mackerel (73.50-99.98%), canned tuna (79.08-103.74%) and salmon (74.56-99.02%). The specificity and method accuracy were expressed as % CV in the range 5.34%-8.48%. Overall, the developed multi-well sensing system (Method 2) showed satisfactory specificity, cost effectiveness, rapidity, and stability for monitoring histamine toxicity as a practical food diagnostic device.

Carvacrol encapsulated nanocarrier/ nanoemulsion abrogates angiogenesis by downregulating COX-2, VEGF and CD31 in vitro and in vivo in a lung adenocarcinoma model.

Nanoemulsion-based synthesis has been introduced to enhance the bioavailability of natural compounds at target sites for their various biomedical applications. In this study, we synthesized carvacrol nanoemulsion (CN) an oil-in-water (O/W) as a nano-emulsion vehicle system by using ultrasonication emulsification for anti-angiogenesis therapy formulated by combining MCT, lecithin, and polysorbate 80 at the O/W interface called carvacrol encapsulated nanoemulsion (CEN). The diameter of CEN determined by TEM analysis was 105.32 nm. The hydrodynamic droplet size was 101.0 nm with a -39.38-mV zeta potential. The stability of the synthesized CEN was approved till 100 days without any change in diameter size distribution and encapsulation efficiency. We evaluated the role of CEN on angiogenesis in lung adenocarcinoma A549 cells both in vitro and in vivo and observed that it reduced the growth and MMP levels of A549 cells in a dose-dependent manner. Exposure to CEN decreased the activation of MAPK p38 as well as ERK. Moreover, we found that CEN reduced the expression of VEGF and CD31 in A549 cells both in vitro and in vivo. Our in-silico study also indicated the binding of carvacrol to COX-2 and VEGF at the active and allosteric sites of CD31 with low binding energy. Overall, CEN induced anti-angiogenic effects in A549 cells in vitro, in silico, and in vivo, thereby establishing its potential as targeted drug delivery vehicle against angiogenesis.

The effect of biogenic manufactured silver nanoparticles on human endothelial cells and zebrafish model.

Human health and environment have been continuously getting exposure to toxic chemicals including nanomaterial; therefore, nontoxicity has recently attracted huge amount of attention. In this study, RU-AgNPs were synthesized by a green synthesis procedure and evaluated for their toxicity in human umbilical vein endothelial cells (HUVECs) as well as on zebrafish embryos via apoptotic pathway. The synthesized RU-AgNPs were average in size (20-25 nm) with a negative surface charge of -13.43 mV. As a result, RU-AgNPs potentiated the formation of reactive oxygen species (ROS) in HUVECs as confirmed by the results of immunoblotting analysis using apoptotic markers, such as Bax, Bcl2, and cytochrome C. Moreover, the induction of apoptosis in HUVECs was also authenticated in a dose-dependent manner after the treatment with RU-AgNPs by the Incucyte analysis. In vivo trials conducted on zebrafish visualized the mortality, malformation, and imbalanced in the heart rate, and cell death of the whole embryo, including severe morphological changes in the yolk sac and the tail of zebrafish. Furthermore, the results of western blot analysis demonstrated the increasing intensity of apoptotic biomarkers such as Bax, Bcl2, and Cyto C, including enhanced production of ROS, validating the cell death in zebrafish larvae. In addition, chemically functionalized silver nanoparticles found to be more cytotoxic than biogenic functionalized silver nanoparticles. Above-mentioned findings clearly demonstrate that Ru-AgNPs cause the toxicity via ROS-induced apoptotic pathway. Therefore, it is necessary to decide RU-AgNPs toxicity levels before being used in any biomedical application.