Krishnan Anand

A549 lung cell line activity of biosynthesized silver nanoparticles using Albizia adianthifolia leaf.

Stable AgNPs were formed in vitro by reacting AgNO3 (aq) solution with the aqueous plant leaf extract. UV-vis revealed the surface plasmon resonance λmax at 448 nm and the absorbance steadily increased in intensity as a function of reaction time. Transmission electron microscope (TEM) and XRD studies were used to characterize the AgNPs; the size was 4-35 nm. Dynamic light scattering (DLS) was used as supporting evidence to determine hydrodynamic size and zeta potential recorded as 80.27 nm and -24.7 mV, respectively. FT-IR spectra suggest that AgNPs are capped with protein molecules and other water soluble phytocompounds such as saponins and glycosides which also behave as stabilizing agents; TEM images indicate a visible layer surrounding the AgNPs. Prominent absorption bands at 3380 and 1642 cm(-1) are assigned to alcohol and carbonyl groups, respectively. (1)H NMR of the neat aqueous plant extract indicates presence of a complex mixture of compounds; however the chemical shift at δ 6.0-8.0 and 1.0-4.0 ppm indicates the presence of few aromatic but abundant aliphatic compounds, respectively. Toxicity of AgNPs on lung cancer cells (A549) and normal healthy peripheral lymphocytes (PLs) at 10 μg/ml and 50 μg/ml was assessed using the MTT, ATP and lactate dehydrogenase assays. Viability data for A549 cells showed a 21% (10 μg/ml) and 73% (50 μg/ml) cell viability after 6h exposure to AgNPs compared to 117% (10 μg/ml) and 109% (50 μg/ml) cell viability of normal peripheral lymphocytes. Lactate dehydrogenase was only significantly altered at 50 μg/ml AgNPs treated cells from 2.43±0.04 units to 0.77±0.04 units.

Silver nanoparticles of Albizia adianthifolia: the induction of apoptosis in human lung carcinoma cell line

BackgroundSilver nanoparticles (AgNP), the most popular nano-compounds, possess unique properties. Albizia adianthifolia (AA) is a plant of the Fabaceae family that is rich in saponins. The biological properties of a novel AgNP, synthesized from an aqueous leaf extract of AA (AAAgNP), were investigated on A549 lung cells. Cell viability was determined by the MTT assay. Cellular oxidative status (lipid peroxidation and glutathione (GSH) levels), ATP concentration, caspase-3/-7, -8 and −9 activities were determined. Apoptosis, mitochondrial (mt) membrane depolarization (flow cytometry) and DNA fragmentation (comet assay) were assessed. The expression of CD95 receptors, p53, bax, PARP-1 and smac/DIABLO was evaluated by flow cytometry and/or western blotting.ResultsSilver nanoparticles of AA caused a dose-dependent decrease in cell viability with a significant increase in lipid peroxidation (5-fold vs. control; p = 0.0098) and decreased intracellular GSH (p = 0.1184). A significant 2.5-fold decrease in cellular ATP was observed upon AAAgNP exposure (p = 0.0040) with a highly significant elevation in mt depolarization (3.3-fold vs. control; p < 0.0001). Apoptosis was also significantly higher (1.5-fold) in AAAgNP treated cells (p < 0.0001) with a significant decline in expression of CD95 receptors (p = 0.0416). Silver nanoparticles of AA caused a significant 2.5-fold reduction in caspase-8 activity (p = 0.0024) with contrasting increases in caspase-3/-7 (1.7-fold vs. control; p = 0.0180) and −9 activity (1.4-fold vs. control; p = 0.0117). Western blots showed increased expression of smac/DIABLO (4.1-fold) in treated cells (p = 0.0033). Furthermore, AAAgNP significantly increased the expression of p53, bax and PARP-1 (1.2-fold; p = 0.0498, 1.6-fold; p = 0.0083 and 1.1-fold; p = 0.0359 respectively).ConclusionData suggests that AAAgNP induces cell death in the A549 lung cells via the mt mediated intrinsic apoptotic program. Further investigation is required to potentiate the use of this novel compound in cancer therapy trials.

Moringa oleifera Gold Nanoparticles Modulate Oncogenes, Tumor Suppressor Genes, and Caspase-9 Splice Variants in A549 Cells.

Gold nanoparticles (AuNPs) facilitate cancer cell recognition and can be manufactured by green synthesis using nutrient rich medicinal plants such as Moringa oleifera (MO). Targeting dysregulated oncogenes and tumor suppressor genes is crucial for cancer therapeutics. We investigated the antiproliferative effects of AuNP synthesized from MO aqueous leaf extracts (MLAuNP) in A549 lung and SNO oesophageal cancer cells. A one‐pot green synthesis technique was used to synthesise MLAuNP. A549, SNO cancer cells and normal peripheral blood mononuclear cells (PBMCs) were exposed to MLAuNP and CAuNP to evaluate cytotoxicity (MTT assay); apoptosis was measured by phosphatidylserine (PS) externalization, mitochondrial depolarization (ΔΨm) (flow cytometry), caspase‐3/7, −9 activity, and ATP levels (luminometry). The mRNA expression of c‐myc, p53, Skp2, Fbw7α, and caspase‐9 splice variants was determined using qPCR, while relative protein expression of c‐myc, p53, SRp30a, Bax, Bcl‐2, Smac/DIABLO, Hsp70, and PARP‐1 were determined by Western blotting. MLAuNP and CAuNP were not cytotoxic to PBMCs, whilst its pro‐apoptotic properties were confirmed in A549 and SNO cells. MLAuNP significantly increased caspase activity in SNO cells while MLAuNP significantly increased PS externalization, ΔΨm, caspase‐9, caspase‐3/7 activities, and decreased ATP levels in A549 cells. Also, p53 mRNA and protein levels, SRp30a (P = 0.428), Bax, Smac/DIABLO and PARP‐1 24 kDa fragment levels were significantly increased. Conversely, MLAuNP significantly decreased Bcl‐2, Hsp70, Skp2, Fbw7α, c‐myc mRNA, and protein levels and activated alternate splicing with caspase‐9a splice variant being significantly increased. MLAuNP possesses antiproliferative properties and induced apoptosis in A549 cells by activating alternate splicing of caspase‐9. J. Cell. Biochem. 117: 2302–2314, 2016.

Sorption isotherms, kinetic and optimization process of amino acid proline based polymer nanocomposite for the removal of selected textile dyes from industrial wastewater

In this article, adsorption and kinetic studies were carried out on three textile dyes, namely Reactive Blue 222 (RB 222), Reactive Red 195 (RR 195) and Reactive Yellow 145 (RY 145). The dyes studied in a mixture were adsorbed under various conditions onto PRO-BEN, a bentonite modified with a new cationic proline polymer (l-proline-epichlorohydrin polymer). The proline polymer was characterized by 1H NMR, Fourier transform infrared spectroscopy (FT-IR), dynamic light scattering (DLS) and TEM. The PRO-BEN composite was characterized by FT-IR, dynamic light scattering (DLS) (zeta potential), TEM imaging, SEM/EDX and X-ray photoelectron spectroscopy (characterize the binding energy). During adsorption studies, factors involving pH, temperature, the initial concentrations of the dyes and the quantity of PRO-BEN used during adsorption were established. The results revealed that the adsorption mechanism was categorized by the Langmuir type 1 isotherm. The adsorption data followed the pseudo-second order kinetic model. The intraparticle diffusion model indicated that adsorption did not only depend on the intraparticle diffusion of the dyes. The thermodynamic parameters verified that the adsorption process was spontaneous and exothermic. The Gibbs free energy values indicated that physisorption had occurred. Successful adsorption of dyes from an industrial effluent was achieved. Desorption studies concluded that PRO-BEN desorbed the dyes better than alumina. This can thereby be viewed as a recyclable remediation material. The PRO-BEN composite could be a cost efficient alternative towards the removal of organic dyes in wastewater treatment.

Design, synthesis, anticancer, antimicrobial activities and molecular docking studies of novel quinoline bearing dihydropyridines

A new series of eight quinoline bearing dihydropyridine derivatives (A1-A8) were synthesized in high yield and in short reaction time by a four component reaction of 2-chloro-3-fomyl quinoline, malononitrile, arylamines and dimethyl acetylenedicarboxylate in the presence of a catalytic amount of triethylamine. The compounds were fully characterized by IR, NMR and GC-MS. These compounds were screened for potential biological activity in an A549 lung cancer cell line and were also evaluated for their antibacterial activities against Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 29213 whilst their molecular docking properties in an enzymatic system were also determined. Compounds A2, A3, A4 and A8 showed anti-proliferative activity; with A4 having the highest toxicity at 250μg/mL and A8 has high toxicity at 125, 250 and 500μg/mL, respectively. Antibacterial results indicated that A4 have significant activity against tested microorganisms at the minimum inhibitory concentration (MIC) values of 32μg/mL against Pseudomonas aeruginosa and Escherichia coli, and 16μg/mL against Staphylococcus aureus. Docking of A1 with human mdm2 indicated the lowest binding energy (-6.111Kcal/mol) thereby showing strong affinity of the ligand molecule with the receptor which has been stabilized by strong hydrogen bond interactions in the binding pocket. This confirms that A1 is a better inhibitor for E3 ubiquitin-protein ligase mdm2.

Cobalt boron nitride: A novel heterogeneous catalyst for the synthesis of medicinally important α-amino quinoline phosphonates

GRAPHICAL ABSTRACT ABSTRACT A novel cobalt supported on boron nitride (CoBNT) heterogeneous catalyst for the synthesis of α-amino quinoline phosphonates (AQPs) is reported in the present work. The CoBNT was synthesised by simply mixing boron nitride in a solution of cobalt acetate, under an inert atmosphere for 7 d followed by filtration; the yield was 94%. It exhibited excellent catalytic properties for the synthesis of 16 novel AQPs in a one pot mixture containing 2-methoxy 3-formyl quinoline, aniline derivatives and diethyl phosphite. Reactions were rapid, products were easily worked-up and were obtained in more than 90% yield. The CoBNT also exhibited higher catalytic activity than conventional catalysts and was re-used five times without significant decrease in catalytic activity.

Cytotoxic Effect of a Novel Synthesized Carbazole Compound on A549 Lung Cancer Cell Line

Increased death rates due to lung cancer have necessitated the search for potential novel anticancer compounds such as carbazole derivatives. Carbazoles are aromatic heterocyclic compounds with anticancer, antibacterial and anti-inflammatory activity. The study investigated the ability of the novel carbazole compound (Z)-4-[9-ethyl-9aH-carbazol-3-yl) amino] pent-3-en-2-one (ECAP) to induce cytotoxicity of lung cancer cells and its mechanism of action. ECAP was synthesized as a yellow powder with melting point of 240-247 °C. The 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), lipid peroxidation and comet assays were used to assess the cytotoxic effect of the compound on A549 lung cancer cells. Protein expression was determined using western blots, apoptosis was measured by luminometry (caspase-3/7, -8 and -9) assay and flow cytometry was used to measure phosphatidylserine (PS) externalisation. ECAP induced a p53 mediated apoptosis of lung cancer cells due to a significant reduction in the expression of antioxidant defence proteins (Nrf2 and SOD), Hsp70 (p < 0.02) and Bcl-2 (p < 0.0006), thereby up-regulating reactive oxygen species (ROS) production. This resulted in DNA damage (p < 0.0001), up-regulation of Bax expression and caspase activity and induction of apoptosis in lung cancer cells. The results show the anticancer potential of ECAP on lung cancer.

Moringa oleifera gold nanoparticles modulate oncogenes, tumor suppressor genes, and Caspase-9 splice variants in A549 cells

Oesophageal cancer ranks the ninth most common malignancy and the sixth most frequent cause of cancer death in the world, with geographic variations in incidence and histological subtypes. In China, oesophageal squamous cell carcinoma (ESCC) accounts for >90% of the total incidence of oesophageal cancer; approximately 300 000 new cases of oesophageal cancer are diagnosed every year in the world of which almost half originate in the high-incidence regions of China.1 According to the Hong Kong Cancer Registry in 2012, ESCC ranked the tenth most frequent cancer death among all other cancers for both sexes. The current treatment modalities for ESCC achieve relatively suboptimal survival and cure rates.2 To further improve the management of this disease, novel prognostic markers for ESCC need to be identified. Gene amplification and overexpression have been suggested to be the major genomic aberrations involved in the pathogenesis of ESCC. Previously, our group reported a novel oncogene JK-1 in ESCC located in the chromosomal region 5p15.1-2; it frequently shows amplification in ESCC and other solid tumours.3 Our collaborators also reported the overexpression of JK-1 mRNA in colorectal tumours, providing the first evidence of the significance of JK-1 mRNA overexpression in gastrointestinal cancer.4 Nonetheless, there are no studies of JK-1 protein expression in ESCC or its correlation with clinicopathological features. Identifying novel histopathologial tumour markers for ESCC is important. Correlating the levels of these protein signals in tissues with the respective clinicopathological features enables better management of the disease. Detection of Hong Kong Med J 2018;24(Suppl 3):S41-4 HMRF project number: 01121886Results: Significant differences in serum leptin levels between the control and the study group were observed. No differences between the pre and post-operative serum leptin levels. Immunohistochemistry studies showed that leptin and its receptors were positively expressed in the majority of breast cancer tissue and this expression was significantly correlated with the distant metastasis of the breast cancer.T aim of the present study is to explore the possible mechanism of ID4 gene in breast tumorigenesis by observing the methylation status of ID4 gene promoter in breast cancer and its relationship with clinical pathological characteristics. The methylation level of ID4 promoter region in breast tumor (n=40) and normal tissue (n=20) specimens was detected by pyrosequencing and the correlation between the methylation level of ID4 gene and clinical pathological characteristics was analyzed. The methylation level of ID4 in MM-453 cell line before and after demethylation treatment was detected by pyrosequencing and the mRNA expression of ID4 was detected by RT-PCR. The methylation level of ID4 promoter region in breast tumor tissue was (31.16±1.5%) and significantly higher than that in normal breast tissue (19.89±0.22%). The methylation level of ID4 gene in ER positive breast cancer tissue was (36.57±1.97%) and significantly higher than that in ER negative group (27.91±1.83%). After the demethylation treatment in MM-453 cell line, the methylation level of ID4 gene decreased and the mRNA expression of ID4 increased remarkably. ID4 gene may play an important role in the breast cancer formation by a variety way including gene promoter hypermethylation, especially in ER positive breast cancer.C cancer is one of the most prevalent and deadly female cancers worldwide and is especially common in developing countries. Various surveys have been conducted in the recent years that revealed various insights into the knowledge, perceptions and practices of the different communities regarding cervical cancer. Studies clearly indicated that women in different communities especially in lower resource settings have limited knowledge about cervical cancer, it’s link with Human Papilloma Virus (HPV), prevention and even less or not at all familiar with screening and HPV vaccine. That lack of awareness may result in poor utilization of HPV vaccine and screening services, even when these are available. The prevention of cervical cancer needs a number of factors to be effectively in place like the awareness of cervical cancer etiology, barriers including intrapersonal, interpersonal and institutional barriers to assessing health care need to be removed, knowledge about the fact that HPV vaccination prevents most of the cervical cancer and screening can detect precancerous lesions which can be mitigated by treatment. Therefore, there is a major need to develop educational intervention programs to address HPV vaccine safety concerns and educate the community by targeting the risk population on risk factors for cervical cancer and practices related to its prevention and early detection. Improvement of cervical health literacy level is a major step towards the prevention and early detection of this dreadful disease.A object segmentation is one of the most important tasks in image analysis. While a universal schema for this task is seemingly not shortly attainable, practical solutions under different circumstances have been developed in the past couple of decades. Most of the previous works were based on machine learning. In contrast, our agenda differs from them in that we try to achieve satisfactory segmentation with a limited number of examples. Our idea is to leverage object matching in the segmentation model, thereby object segmentation is automated or object matching becomes precise in pixel level. In this talk, I’ll introduce two of my recent works in this direction. Specifically, I’ll introduce how object matching model can be integrated with two different types of segmentation models, namely the Markov random field model and the active contour model, to achieve automatic object segmentation. Discussions on their uses in radiographic image analysis would be encouraged.L biopsies are non-invasive blood diagnostic tests that detect circulating tumor cells (CTCs) and/or fragments of tumor DNA that are shed into the blood from the primary tumor and from metastatic sites. This approach can have an enormous diagnostic and treatment implication for oncology that can transform clinical oncology practice and that it is part of the personalized medicine. Whereas tumor genome sequencing is already central to inform treatment decisions and the management of oncological patients, the liquid biopsy may represent the non-invasive approach to monitor tumor genomic changes in real time. This will allow clinicians to ensure that the therapy they have selected based on a particular molecular target, remains relevant and eventually observe the emergence of any resistance. Eventually, it would be possible to observe if any new molecular targets appear that could be suitable for a different treatment. All this could help to provide patients with the right treatment for the right target without delay. Liquid biopsies also present us with a unique opportunity to move forward with our understanding of metastatic disease development and they may help to identify signaling pathways involved in cell invasiveness and metastatic competence. Moreover these tests have the possibility to be used in screening programs at least for some kind of cancers. At the end, the liquid biopsy can revolutionize cancer care, providing clinicians with rapid access to information on a molecular level at diagnosis, thereby optimizing treatment choices.B circulating microRNAs are known as novel and validated biomarkers for application in early detection of diseases including cancers. The aim was to design and fabricate precise sensors for quantification of miRNAs in real samples (serum/plasma) without any sample manipulation. Two sensors (an electrochemical biosensor and nanobiosensor) were developed for quantification of miR-155, as a biomarker related to the early phase of breast cancer expression. Both sensors showed high specificity and selectivity, so that they could discriminate between target miR-155 and single-base mismatch, three-base mismatch, completely unmatched oligonucleotides. Reproducibility, stability and functionality of the sensors in the real samples (serum/plasma) were found to be significant. Both sensors had a low detection limit and wide linear ranges with simplicity, less consumed time and low cost, which were superior to the most of previously published works. It seems our method can be introduced to be used in clinical labs and also detection of other miRNA biomarkers through other optimizations.