Mycochemistry, antioxidant content, and antioxidant potentiality of the ethanolic extract of Pleurotus florida and its anti-cancerous effect on HeLa cancer cell line, and antitumor effect on HeLa-implanted mice.

Objectives: Cervical cancer is increasing worldwide and is becoming resistant to the existing drugs in clinical practice. Here, ethanolic extract of fruit body of Pleurotus florida was evaluated as antioxidant, anticancer agent against HeLa cell lines and anti-tumor against cervical cancer in mice model. Methods: Fruit bodies of P. florida in 90% ethanol, and the P. florida ethanolic extract (PFEE) was subsequently investigated for its antioxidant content and activity, anticancer properties against the cervical cancer cell line, HeLa, and antitumor activity against HeLa implanted mice. Results: The antioxidant activity bioassay showed that the IC50 of PFEE was 41.17 ± 1.42a µg/ml. The cytotoxicity assay revealed that PFEE caused inhibition of cell proliferation. At the highest dose (1,250 µg/ml) after 24 h, 48 h, or 72 h of treatment, the percentages of cell growth inhibition were 75.22%, 77.77%, and 84.65%, respectively. It revealed that PFEE-treated cells became rounded and the nuclei became fragmented. PFEE induced intracellular generation of reactive oxygen species and reduced the mitochondrial membrane potential. PFEE also led to an up regulation of the apoptotic genes for caspases-3, -9, and Bax, whereas Bcl-2 gene was down regulated, and it also promoted the expression of p53. Cell cycle analysis revealed that cell cycle was arrested at the G0/G1 checkpoint. PFEE suppressed metastasis and colonization. At a dosage of PFEE of 50 mg/kg of body weight, a 66.72% reduction in the size of tumors and an 87.44% reduction in the tumor weight were observed in mice. Conclusions: It has demonstrated that PFEE is a highly potent anti-cervical cancer agent in vitro and in vivo.

Accurate species identification of food-contaminating beetles with quality-improved elytral images and deep learning.

Food samples are routinely screened for food-contaminating beetles (i.e., pantry beetles) due to their adverse impact on the economy, environment, public health and safety. If found, their remains are subsequently analyzed to identify the species responsible for the contamination; each species poses different levels of risk, requiring different regulatory and management steps. At present, this identification is done through manual microscopic examination since each species of beetle has a unique pattern on its elytra (hardened forewing). Our study sought to automate the pattern recognition process through machine learning. Such automation will enable more efficient identification of pantry beetle species and could potentially be scaled up and implemented across various analysis centers in a consistent manner. In our earlier studies, we demonstrated that automated species identification of pantry beetles is feasible through elytral pattern recognition. Due to poor image quality, however, we failed to achieve prediction accuracies of more than 80%. Subsequently, we modified the traditional imaging technique, allowing us to acquire high-quality elytral images. In this study, we explored whether high-quality elytral images can truly achieve near-perfect prediction accuracies for 27 different species of pantry beetles. To test this hypothesis, we developed a convolutional neural network (CNN) model and compared performance between two different image sets for various pantry beetles. Our study indicates improved image quality indeed leads to better prediction accuracy; however, it was not the only requirement for achieving good accuracy. Also required are many high-quality images, especially for species with a high number of variations in their elytral patterns. The current study provided a direction toward achieving our ultimate goal of automated species identification through elytral pattern recognition.

Optimized imaging methods for species-level identification of food-contaminating beetles.

Identifying the exact species of pantry beetle responsible for food contamination, is imperative in assessing the risks associated with contamination scenarios. Each beetle species is known to have unique patterns on their hardened forewings (known as elytra) through which they can be identified. Currently, this is done through manual microanalysis of the insect or their fragments in contaminated food samples. We envision that the use of automated pattern analysis would expedite and scale up the identification process. However, such automation would require images to be captured in a consistent manner, thereby enabling the creation of large repositories of high-quality images. Presently, there is no standard imaging technique for capturing images of beetle elytra, which consequently means, there is no standard method of beetle species identification through elytral pattern analysis. This deficiency inspired us to optimize and standardize imaging methods, especially for food-contaminating beetles. For this endeavor, we chose multiple species of beetles belonging to different families or genera that have near-identical elytral patterns, and thus are difficult to identify correctly at the species level. Our optimized imaging method provides enhanced images such that the elytral patterns between individual species could easily be distinguished from each other, through visual observation. We believe such standardization is critical in developing automated species identification of pantry beetles and/or other insects. This eventually may lead to improved taxonomical classification, allowing for better management of food contamination and ecological conservation.

Anticancer Activity of Solvent Extracts of Hexogonia glabra against Cervical Cancer Cell Lines.

OBJECTIVE: In this study, we aimed to harness some solvent extracts of one wild mushroom Hexagonia glabra and test their anti-cancer activity against cervical human cell lines, namelyHeLa, SiHa, and CaSki. METHODS: It includes cell morphological study by microscope, nuclear morphology by DAPI staining under fluorescence microscopy, apoptosis assay by fluorescence technique, anti-proliferation by MTT assay and expression of apoptotic and anti-apoptotic genes by Western blotting and cell cycle analysis was done. RESULTS: The selected cervical cancer cells were treated separately with 150 µg/mL of three extracts, namely of ethanolic (EE), ethyl acetate (EAE), and water extract (WE) and exhibited features like round, shrink and dead. All extracts caused apoptosis in cell lines and EE had the highest effect in this regard. The percentages of apoptotic cells in HeLa, SiHa and CaSki, at the same concentration of EE were 79.23, 75.42, and 76.36% respectively. Cytotoxicity assay showed that all three extracts (50 - 250 µg/mL) were potent for inhibition of cell growth of three cell lines and again EE had the highest effect. The percentages of cell growth inhibition in HeLa, SiHa, and CaSki cells treated with EE at 24 h at 50 µg/mL were 45.79±4.11, 41.66±4.03, and 36.72±2.67, while they were 74.23±7.45, 62.31±5.97, and 54.23±5.04 at 150 µg/mL concentration. At 250 µg/mL concentration, the percentages of cell growth inhibition were 94.25 ±8.11, 90.02 ±8.67, and 85.43±6.21, respectively. The expression of apoptotic gene (Caspase 3, 9) and tumor guard gene (p53), as their proteins in Western blotting increased . However, anti-apoptotic BcL2 gene of all cell lines was decreased following treatment with extracts. In addition, the cell cycle analysis (CaSki cell) showed that treatment (EE) arrested at G2/M check point cell cycle. CONCLUSION: All extracts of this mushroom were active in arresting growth of three cell lines and EE had the highest effect, indicating that this mushroom can be a valuable source of anticancer agents.

Estimating Bacterial Concentrations in Fibrous Substrates Through a Combination of Scanning Electron Microscopy and ImageJ.

Conventional signal-based microanalytical techniques for estimating bacterial concentrations are often susceptible to false signals. A visual quantification, therefore, may compliment such techniques by providing additional information and support better management decisions in the event of outbreaks. Herein, we explore a method that combines electron microscopy (EM) and image-analysis techniques and allows both visualization and quantification of pathogenic bacteria adherent even to complex nonuniform substrates. Both the estimation and imaging parameters were optimized to reduce the estimation error ( E, %) to close to ±5%. The method was validated against conventional microbiological techniques such as the use of optical density, flow cytometry, and quantitative real-time PCR (qPCR). It could easily be tailored to estimate different species of pathogens, such as Escherichia coli O157, Listeria innocua, Staphylococcus aureus, Enterococcus faecalis, and Bacillus anthracis, on samples similar to those in real-time contamination scenarios. The present method is sensitive enough to detect ∼100 bacterial CFU/mL but has the potential to estimate even lower concentrations with increased imaging and computation times. Overall, this imaging-based method may greatly complement any signal-based pathogen-detection technique, especially in negating false signals, and therefore may significantly contribute to the field of analytical microbiology and biochemistry.

Comparing SVM and ANN based Machine Learning Methods for Species Identification of Food Contaminating Beetles.

Insect pests, such as pantry beetles, are often associated with food contaminations and public health risks. Machine learning has the potential to provide a more accurate and efficient solution in detecting their presence in food products, which is currently done manually. In our previous research, we demonstrated such feasibility where Artificial Neural Network (ANN) based pattern recognition techniques could be implemented for species identification in the context of food safety. In this study, we present a Support Vector Machine (SVM) model which improved the average accuracy up to 85%. Contrary to this, the ANN method yielded ~80% accuracy after extensive parameter optimization. Both methods showed excellent genus level identification, but SVM showed slightly better accuracy  for most species. Highly accurate species level identification remains a challenge, especially in distinguishing between species from the same genus which may require improvements in both imaging and machine learning techniques. In summary, our work does illustrate a new SVM based technique and provides a good comparison with the ANN model in our context. We believe such insights will pave better way forward for the application of machine learning towards species identification and food safety.

Liquid Crystal Elastomer Microspheres as Three-Dimensional Cell Scaffolds Supporting the Attachment and Proliferation of Myoblasts.

We report that liquid crystal elastomers (LCEs), often portrayed as artificial muscles, serve as scaffolds for skeletal muscle cell. A simultaneous microemulsion photopolymerization and cross-linking results in nematic LCE microspheres 10-30 µm in diameter that when conjoined form a LCE construct that serves as the first proof-of-concept for responsive LCE muscle cell scaffolds. Confocal microscopy experiments clearly established that LCEs with a globular, porous morphology permit both attachment and proliferation of C2C12 myoblasts, while the nonporous elastomer morphology, prepared in the absence of a microemulsion, does not. In addition, cytotoxicity and proliferation assays confirm that the liquid crystal elastomer materials are biocompatible promoting cellular proliferation without any inherent cytotoxicity.

Protein-induced configuration transitions of polyelectrolyte-modified liquid crystal droplets.

Liquid crystal (LC) droplets dispersed in aqueous solution have emerged as an optical probe for sensing the adsorption and interaction of biological species at the LC/aqueous interface. In this paper, we modify the surface of 4-n-pentyl-4'-cyanobiphenyl (5CB) LC droplets by the adsorption of positively charged poly(diallyldimethylammonium chloride) (PDADMAC) and poly(ethylenimine) (PEI) with different molecular weights at the 5CB/water interface. The PDADMAC and PEI-modified 5CB droplets show a radial director configuration in aqueous solution with salt concentrations above 150 mM. The adsorption of negatively charged bovine serum albumin (BSA) on the positively charged PDADMAC and PEI-modified 5CB droplets through electrostatic interaction can induce the radial-to-bipolar configuration transition of the 5CB inside the droplets. We find that the concentration of BSA required to induce the configuration transition increases linearly with the decrease of the molecular weight of PDAMAC and PEI. Our results highlight the capability of the director configuration of LC droplets as an optical probe for sensing the interaction between proteins and polyelectrolytes at the LC/aqueous interface.

Optical detection of lithocholic acid with liquid crystal emulsions.

The concentration level of bile acids is a clinical biomarker for the diagnosis of intestinal diseases because individuals suffering from intestinal diseases have a sharply increased concentration of bile acids at micromolar levels. Here, we report the detection of lithocholic acid (LCA) in aqueous solution by using surfactant-stabilized 4-n-pentyl-4'-cyanobiphenyl (5CB) liquid crystal droplets as an optical probe. We find that the surfactant adsorbed at the 5CB/water interface can be replaced by LCA, triggering a radial-to-bipolar configuration transition of the 5CB in the droplets. By simply observing the LCA-triggered transition with a polarizing optical microscope, micromolar levels of LCA in aqueous solution can be detected. The detection limit and selectivity of surfactant-stabilized 5CB droplets for LCA depend on the chain length and headgroup of the surfactants used for stabilizing 5CB droplets.

Longitudinal zipping/unzipping of self-assembled organic tubes.

Stimuli-responsive organic tubes are an attractive supramolecular assembly which has potential applications as a controlled release vehicle. We synthesize a smart organic tube by the coassembly of lithocholic acid (LCA) and taurolithocholic acid (TLCA) in aqueous solution. The coassembled LCA/TLCA tubes can be longitudinally unzipped into flat sheets by capillary force after being dehydrated on substrates. Consequently, the encapsulated guest molecules are released from the unzipping tubes. After the release of guest molecules, the flat sheets can be zipped back into hollow tubes upon hydration with aqueous solution. The zipping/unzipping LCA/TLCA tubes provide a new type of delivery vehicles, which may have potential for surface decontaminations.

Study of mechanism of enhanced antibacterial activity by green synthesis of silver nanoparticles.

The extensive use of silver nanoparticles needs a synthesis process that is greener without compromising their properties. The present study describes a novel green synthesis of silver nanoparticles using Guava (Psidium guajava) leaf extract. In order to compare with the conventionally synthesized ones, we also prepared Ag-NPs by chemical reduction. Their optical and morphological characteristics were thoroughly investigated and tested for their antibacterial properties on Escherichia coli. The green synthesized silver nanoparticles showed better antibacterial properties than their chemical counterparts even though there was not much difference between their morphologies. Fourier transform infrared (FTIR) spectroscopic analysis of the used extract and as-synthesized silver nanoparticles suggests the possible reduction of Ag(+) by the water-soluble ingredients of the guava leaf like tannins, eugenol and flavonoids. The possible reaction mechanism for the reduction of Ag(+) has been proposed and discussed. The time-dependent electron micrographs and the simulation studies indicated that a physical interaction between the silver nanoparticles and the bacterial cell membrane may be responsible for this effect. Based on the findings, it seems very reasonable to believe that this greener way of synthesizing silver nanoparticles is not just an environmentally viable technique but it also opens up scope to improve their antibacterial properties.

Director configuration transitions of polyelectrolyte coated liquid-crystal droplets.

Liquid-crystal droplets are of great interest because of their large surface areas, rich phases, and tunable optical properties. The director configuration of liquid-crystal droplets provides a unique optical sign to detect the events occurring at the droplet surface. In this article, we report the alternating bipolar/radial configuration transitions of 4-n-pentyl-4'-cyanobiphenyl (5CB) droplets triggered by the layer-by-layer coating of negatively charged poly(styrenesulfonate sodium (PSS) and positively charged poly(diallyldimethylammonium chloride) (PDADMAC) on the droplet surface. The alternating configuration transitions are due to the interactions of the 5CB with polar versus nonpolar PDADMAC/PSS multilayer coatings. Furthermore, we find that the coating of PDADMAC/PSS multilayers makes the director configuration of the 5CB in the droplets sensitive to environmental salt concentrations.

Boojum and stripe textures in long-range orientationally ordered monolayers on solid substrates.

Long-range organization of molecular tilt azimuth is a striking feature in monolayers at the air-water interface. We show that the boojum and stripe textures of pentadecanoic acid (PDA) with the continuous variations of molecular tilt azimuth formed at the air-water interface at temperatures lower than room temperature can be preserved after being transferred to glass substrates at low dipping speeds. The long-range tilt order in the transferred boojums and stripes is resolved by frictional force microscopy at room temperature, suggesting that the tilt order is "frozen" through the interaction of PDA molecules with the glass surface. The transferred stripe structure can be used as a unique alignment layer to induce a continuously azimuthal orientation of nematic liquid crystals.

MWCNTs as reinforcing agent to the hap-gel nanocomposite for artificial bone grafting.

The essence of this investigation is to explore MWCNTs as reinforcing agents to strengthen Hap-Gel nanocomposites for artificial bone grafting applications without significantly compromising their biocompatibility. Hap-Gelatin composites, reinforced with various proportions of MWCNTs, were synthesized to optimize the MWCNT content in the composites which yield commendable improvement in the strength. The morphological studies reveal that the MWCNTs act as templates for nucleation of Hap crystals. The biocompatibility of MWCNT reinforced Hap-Gelatin composites were evaluated in animal model through the histopathological investigation of tissues from skin, kidney, and liver. On histopathological examination, no noticeable alteration due to toxicity was found for lower concentration of MWCNTs. Mild reversible changes in the liver and tubular damage in kidney have been observed for higher concentration (4 wt % of MWCNTs). It can be inferred from the findings that MWCNTs, in proportions less than 4%, can successfully be used to reinforce the Hap-Gel nanocomposite to improve its mechanical properties. However, how safe would these CNT reinforced bone implants would be when used for prolonged period in actual physiological conditions needs to be investigated further.

Characterization of antiplatelet properties of silver nanoparticles.

Thrombotic disorders have emerged as serious threat to society. As anticoagulant and thrombolytic therapies are usually associated with serious bleeding complications, the focus has now shifted to regulating and maintaining platelets in an inactive state. In the present study we show that nanosilver has an innate antiplatelet property and effectively prevents integrin-mediated platelet responses, both in vivo and in vitro, in a concentration-dependent manner. Ultrastructural studies show that nanosilver accumulates within platelet granules and reduces interplatelet proximity. Our findings further suggest that these nanoparticles do not confer any lytic effect on platelets and thus hold potential to be promoted as antiplatelet/antithrombotic agents after careful evaluation of toxic effects.

Characterization of biomimetically synthesized Hap-Gel nanocomposites as bone substitute.

There is an increasing demand for an affordable and easy-to-fabricate material to help patients having a long bone gap. In this paper, we describe the biomimetic synthesis of Hap-Gel in situ nanocomposite powders with varied proportions. Their biocompatibility and bone regeneration abilities were assessed on a rabbit model. The use of Hap crystals and Gel molecule, the soluble form of bone protein, makes the nanocomposites comparable to natural bone in constituents. The application of biomimetic principles improves crystal morphology and the interaction of Hap crystals with the Gel molecules as seen through in vitro characterizations. Out of the various compositions studied, one with 80:20 proportions of Hap to Gel proved to be closest to the characteristics of natural bone. The immunological response to this composite, assessed through intradermal inoculation, did not reveal any reaction. The in vivo implantation studies in the femoral condyle of the animals, as assessed by serial post-operative follow-up radiography and the histological evaluation, revealed a good biocompatibility and bone-regeneration ability of the material. Thus, nanocomposites of Hap-Gel have a great potential for serving as an effective and affordable biomaterial for bone grafting applications.

Retracted: Characterization of enhanced antibacterial effects of novel silver nanoparticles.

In the present study, we report the preparation of silver nanoparticles in the range of 10-15 nm with increased stability and enhanced anti-bacterial potency. The morphology of the nanoparticles was characterized by transmission electron microscopy. The antibacterial effect of silver nanoparticles used in this study was found to be far more potent than that described in the earlier reports. This effect was dose dependent and was more pronounced against gram-negative bacteria than gram-positive organisms. Although bacterial cell lysis could be one of the reasons for the observed antibacterial property, nanoparticles also modulated the phosphotyrosine profile of putative bacterial peptides, which could thus affect bacterial signal transduction and inhibit the growth of the organisms.