The H9c2(2-1) cell-based sulforhodamine B assay is a non-animal alternative to evaluate municipal wastewater quality over time.

The present study validates the potential of the in vitro H9c2(2-1) cell-based sulforhodamine B (SRB) assay to evaluate the temporal variability of wastewater quality. The impact of effluent disposal on water quality and the efficiency of the wastewater treatment process were also assessed. To correlate standard analytical method results with in vitro results, a total of 16 physicochemical parameters, such as nutrients, pH, chemical oxygen demand, total suspended solids and metals, were determined in both raw and treated wastewater samples. Results revealed that the H9c2(2-1) cell-based SRB assay has an enormous potential to evaluate municipal wastewater quality over time and to discriminate influent and effluent toxic characteristics, as well as for water quality monitoring and surveillance of the efficacy of treatment processes. Finally, the gathered results alerted to the impact of phosphates in a biological system, leading us to recommend the selection of this parameter as a potential environmental health indicator.

Anticancer activity of gold nanobioconjugates synthesized from Elephantopus scaber (linn.) leaf extract.

Introduction: Medicinal plants are the major natural resources for the treatment of human ailments including cancer therapy. The current cancer treatments such as surgery, radiation, and chemotherapy affect normal cells too. Thus, treatments like synthesized nanoscale particles using plant extracts have proven to be potential anticancer agent. Aim of the Study: We hypothesize that the gold nanoparticles (AuNPs) synthesized using Elephantopus scaber hydro-methanolic extract may have anti-cancer activity along with their synergistic counterparts with adriamycin (ADR) on human breast cancer MCF-7: human breast cancer (A-549), human oral cancer (squamous cell carcinoma [SCC]-40), and COLO-205: human colon cancer cell lines. Materials and Methods: The phytosynthesized AuNPs were characterized for ultraviolet-visible (UV-Vis) spectroscopy, nanoparticle tracking analysis (NTA), X-ray diffraction, scanning electron microscopy, transmission electron microscopy (TEM), and Fourier transform infrared (FTIR) analysis. The anticancer ability of the AuNPs against human MCF-7, A-549, SCC-40, and COLO-205 through sulforhodamine B assay has been studied. Results: The synthesis of AuNPs was confirmed with the UV-Vis spectrophotometer with a peak at 540 nm. The FTIR analysis showed polyphenolic groups were major found to be the reduction and capping agent for AuNPs. According to the results obtained, AuNPs showed good anti-proliferative activity with GI50 <10 µg/ml on MCF-7 cancer cell line. The synergistic effect of AuNPs + ADR was even better for all the four cell lines than that of the AuNPs alone. Conclusion: The green synthesis of AuNPs is a simple, eco-friendly, and cost-effective method with dominantly spherical morphology ranging from 20 to 40 nm confirmed by NTA and TEM analysis. The study reveals the potent therapeutic value of the AuNPs.

A3 Adenosine Receptor Antagonists with Nucleoside Structures and Their Anticancer Activity.

The overexpression of the A3 adenosine receptor (AR) in a number of cancer cell types makes it an attractive target for tumor diagnosis and therapy. Hence, in the search for new A3AR ligands, a series of novel 2,N6-disubstituted adenosines (Ados) was synthesized and tested in radioligand binding and functional assays at ARs. Derivatives bearing a 2-phenethylamino group in the N6-position were found to exert higher A3AR affinity and selectivity than the corresponding N6-(2,2-diphenylethyl) analogues. 2-Chloro-N6-phenylethylAdo (15) was found to be a potent full A3AR agonist with a Ki of 0.024 nM and an EC50 of 14 nM, in a cAMP accumulation assay. Unlike 15, the other ligands behaved as A3AR antagonists, which concentration-dependently reduced cell growth and exerted cytostatic activity on the prostate cancer cell line PC3, showing comparable and even more pronounced effects with respect to the ones elicited by the reference full agonist Cl-IB-MECA. In particular, the N6-(2,2-diphenylethyl)-2-phenylethynylAdo (12: GI50 = 14 µM, TGI = 29 µM, and LC50 = 59 µM) showed the highest activity proving to be a potential antitumor agent. The cytostatic effect of both A3AR agonist (Cl-IB-MECA) and antagonists (12 and other newly synthesized compounds) confirm previous observations according to which, in addition to the involvement of A3ARs, other cellular mechanisms are responsible for the anticancer effects of these ligands.

Synthesis of New Pyrazole Hybrids as Potential Anticancer Agents with Xanthine Oxidase Inhibitory Activity.

AIMS: The study aims to synthesize hybrid molecules containing pyrazole and aryldiazenyl/arylhydrazono fragments with promising anticancer activity. BACKGROUND: The clinical effectiveness of anticancer drugs is limited by their adverse side effects and patient resistance. Therefore, the development of safer classes of drugs through rational drug design is imperative. OBJECTIVE: Considering the anticancer potential of the pyrazole moiety, the study was carried out with the objective of synthesizing some hybrid pyrazole derivatives with anticancer potential. METHODS: The anticancer potential of these pyrazolyl analogues were evaluated by sulforhodamine B assay using three cancer cell lines MCF-7, HepG2, and HCT-116. RESULTS: HCT-116 was the most sensitive cell line against these pyrazolyl analogues. Among these newly synthesised derivatives, 1-(4-((4-bromophenyl)diazenyl)-3,5-dimethyl-1H-pyrazol-1-yl)-2-(naphthalen-2-yloxy)ethan-1-one (5e) emerged as a promising anticancer agent (IC50 3.6-24.6 µM), having a xanthine oxidase inhibitory effect (IC50 10.87 µM). To obtain further insights into the binding interactions of these molecules, molecular docking studies were also carried out. CONCLUSION: In summary, our findings suggest that these hybrid pyrazolyl derivatives can be considered as potential lead molecules for anticancer agents.

Anti-EGFR-mAb and 5-Fluorouracil Conjugated Polymeric Nanoparticles for Colorectal Cancer.

BACKGROUND: Due to the higher intake of junk food and unhealthy lifestyle, the percentage of U.S. adults aged 50 to 75 years who were up-to-date with colorectal cancer screening increased 1.4 percentage points, from 67.4% in 2016 to 68.8% in 2018. This represents an additional 3.5 million adults screened for colorectal cancer. This is a severe concern of this research, and an attempt was made to prepare a target-specific formulation that could circumvent chemotherapy-related compilation and improvise higher cellular uptake. The fundamental agenda of this research was to prepare and develop Anti-EGFR mAb and 5-Fluorouracil (5-FU) fabricated polymeric nanoparticles for colorectal cancer. OBJECTIVE: The main objective of this research was to prepare and evaluate more target specific formulation for the treatment of colorectal cancer. PLGA and PEG-based polymeric nanoparticles are capable of preventing opsonization via the reticuloendothelial system. Hence, prepared polymeric nanoparticles are capable of higher cellular uptake. METHODS: The Poly(d,1-lactide-co-glycolide) (PLGA) and Polyethylene Glycol (PEG) were combined utilizing the ring-opening polymerization method. The presence of PEG prevents opsonization and distinguished blood concentration along with enhanced targeting. The presence of PLGA benefits in the sustained release of polymeric formulations. The optimized formulation (5-FU-PLGA- PEG-NP) was lyophilized using 4% trehalose (cryoprotectants) and conjugated with Anti- EGFR mAb on its surface to produce Anti-EGFR-5-FU-PLGA-PEG-NP; the final formulation, which increases target specificity and drug delivery system of nanoparticles. RESULTS: The spherical shaped optimized formulation, 5-FU-PLGA-PEG-NP-3 was found to have higher percentage drug entrapment efficacy (71.23%), higher percentage drug content (1.98 ± 0.34%) with minimum particles size (252.3nm) and anionic zeta potential (-31.23mV). The IC50 value of Anti-EGFR-5-FU-PLGA-PEG-NP was 1.01µg/mL after 48 hours incubation period in the HCT 116 cell line, indicating higher anticancer effects of the final formulation. CONCLUSION: From the outcomes of various experiments, it was concluded that Anti-EGFR-5-FUPLGA- PEG-NP has biphasic drug release kinetics, higher cellular uptake and higher cytotoxicity. Therefore, anti-EGFR-5-FU-PLGA-PEG-NP holds excellent potential for drug delivery to EGFR positive colorectal cancer cells.

Purification, characterization and anticancer evaluation of l-methioninase from Trichoderma harzianum.

The Trichoderma harzianum l-methioninase was purified 7.15-fold with a recovery of 47.9% and the specific activity of 74.4 U/mg of protein. The purified enzyme has an apparent molecular mass of 48 kDa on SDS-PAGE and exhibited maximum activity at pH 8 and 35 °C. The enzyme was catalytically stable below 50 °C and at a pH range of 6.0-8.5. The thermal inactivation of l-methioninase exhibited first-order kinetics with the k value between 5.71 × 10-4 min-1 and 1.83 × 10-2 min-1. The studies on thermodynamic parameters of l-methioninase indicated the compaction and aggregation of the enzyme molecule during denaturation. This is the first report of thermodynamic analysis of thermal inactivation in l-methioninase. The enzyme activity was enhanced by Li+ and inhibited by Cu2+, Co2+, Fe2+, Hydroxylamine and PMSF. The purified enzyme showed K m , V max and k cat value of 1.19 mM, 21.27 U/mg/min and 16.11 s-1, respectively. The l-methioninase inhibited the growth of human cell lines hepatocellular carcinoma (Hep-G2) and breast carcinoma (MCF-7) with IC50 values of 14.12 µg/ml and 20.07 µg/ml, respectively. The in vivo antitumor activity of l-methioninase was evaluated against DAL cell lines bearing in Swiss albino mice. The enzyme effectively reduced tumor volume, packed cell volume, viable cell count and restored hematological parameters, serum enzyme and lipid profile to normal levels compared to DAL control mice. The present study has demonstrated the high efficacy of Trichoderma harzianum l-methioninase against cancer cell lines in vitro and in vivo conditions. The purified l-methioninase has significant thermal stability and better catalytic properties than the enzyme purified from other sources.

Antiproliferative activity of Camellia sinensis mediated silver nanoparticles on three different human cancer cell lines.

INTRODUCTION: Cancer is one of the leading causes of mortality in the world and there are many types of cancer. The current treatments against cancer are surgery, chemotherapy, and radiation therapy, but these come with varied side effects as they harm noncancer cells too. Therefore, search for new treatments is one of the important research areas nowadays and nanoparticles are one such potential anticancer agent. AIM OF THE STUDY: We hypothesized that silver nanoparticles and tea extract may have anticancer activities along with their synergistic counterparts with adriamycin (ADR) on HT-29 human colon cancer cell line, MCF-7 human breast cancer cell line, and MOLT-4 human leukemia cancer cell line. MATERIALS AND METHODS: The biosynthesized silver nanoparticles were characterized by ultraviolet-visible spectroscopy, nanoparticle tracking analyzer (NTA), X-ray diffraction, scanning electron microscopy, transmission electron microscopy (TEM), and Fourier transform infrared analysis. The cytotoxic activity was measured using the sulforhodamine B assay protocol on the HT-29, MCF-7, and MOLT-4 cell lines. RESULTS: The synthesized AgNPs gave absorption maxima at 415 nm, with four different diffraction peaks (°2θ values) corresponding to the face centered cubic silver lines. Our results showed that AgNPs exhibited the highest cytotoxic activity at 20 µg/mL concentration against all the three cell lines followed by the combination of AgNPs+ADR. CONCLUSION: The superior activity of the silver nanoparticles may be due to its spherical shape and smaller particle size 10-30 nm as confirmed from NTA and TEM analysis. The data obtained in the study reveal the potent therapeutic value of biogenic silver nanoparticles.

Less-toxic rearrangement products of NX-toxins are formed during storage and food processing.

A new type A trichothecene mycotoxin, NX-2, was previously reported to be produced by North American isolates of the cereal pathogen Fusarium graminearum. Here we describe the isolation and structural characterization of a rearrangement product, called NX2-M1, and related compounds with different acetylation patterns (NX3-M1 and NX4-M1). In the NX-M1 derivatives, the epoxide ring is opened, and a covalent bridge between C-10 and C-12 of the trichothecene backbone is formed. In vitro translation assays showed that NX3-M1 is less toxic for eukaryotic ribosomes than NX-3. NX3-M1 also has a greatly reduced cytotoxic potential on two tested human colon cell lines. Formation of NX3-M1 can therefore be regarded as a detoxification reaction. The related F. graminearum mycotoxin deoxynivalenol (DON), which is frequently occurring worldwide, is very stable during food processing. Testing NX-3 at different pH-values and temperature conditions, as well as under conditions that simulate the storage of infected grains and bread-making process, revealed a strongly reduced stability of NX-3 and concurrent formation of NX3-M1. Although the NX-3 formed in planta is as toxic as DON, the extensive formation of the non-toxic rearrangement product should be taken into account for risk assessment of this emerging food contaminant.

Synthesis of 17ß-N-arylcarbamoylandrost-4-en-3-one derivatives and their anti-proliferative effect on human androgen-sensitive LNCaP cell line.

In this study, we report the synthesis and anti-proliferative effect of a set of eight androst-4-ene-3-one derivatives with different arylcarbamoyl groups at C-17. The novel compounds were prepared from commercially available 3ß-hydroxy-5-pregnen-20-one and evaluated against the androgen-sensitive human prostate adenocarcinoma LNCaP cell line. The cancerous cells were exposed to 50 µM of each compound and the proliferating agent testosterone (T) or dihydrotestosterone (DHT). The most potent compounds from this assay were further tested against the androgen-insensitive PC3 cell line. We also demonstrate the activity of these compounds on rat peripheral blood mononuclear cells for comparison. Both 17ß-N-[3,5-bis(trifluoromethyl)phenylcarbamoyl]androst-4-ene-3-one (6f) and 17ß-N-(1,3-thiazol-2-ylcarbamoyl)androst-4-ene-3-one (6g) exhibited a higher growth inhibitory effect than commercially available drugs finasteride, flutamide and ketoconazole on LNCaP cells in the presence and absence of androgens. In addition, 6f and 6g demonstrated high potency on PC3 cells suggesting an androgen-independent anti-proliferative effect. Moreover, the novel compounds showed a small effect on rat mononuclear cells, an indication of low toxicity.

Synthesis, characterization and in vitro evaluation of methotrexate conjugated fluorescent carbon nanoparticles as drug delivery system for human lung cancer targeting.

Nanotechnology based cancer therapeutics have rapidly advanced towards the solution of many limitations associated with other drug delivery agents such as nonspecific distribution within the body, low water solubility and non-biocompatibility. Carbon nanoparticles have demonstrated unique properties that are useful to combat with these issues, including their properties dependent on size, high stability in different solvents, compatible size for drug delivery and ease of surface modifications. Fluorescent carbon nanoparticles with good water solubility were obtained from a carbohydrate source by acid assisted ultrasonic treatment at 35kHz for 4h. This simple and economical method can be used for large scale production. Electron microscopic, spectroscopic and thermo gravimetric analysis techniques were used to characterize these carbon nanoparticles. Functionalized CNPs were further conjugated with anticancer drug-methotrexate and used as fluorescent nano-carriers. In this research work, we determined the in vitro bioactivity of CNPs-methotrexate conjugates by lactate dehydrogenase assay, cell adhesion assay and sulforhodamine B assay in human lung carcinoma cell line (H157). The CNPs showed promising biocompatibility and CNPs-MTX conjugates demonstrated potent cytotoxic effects and high anticancer activities in human lung cancer cell line.