Mouse Embryo Fibroblast Cell Line Research Articles

Phytoactive essential oils-composed water-free organogels: Development, characterization and proof of antibacterial activity

The objective of this study was to develop and analyze a gel-like formulation that does not contain water, made from natural compounds found in essential oils, which are known for their antimicrobial properties and ability to reduce inflammation. Lentisk tree (Pistacia lentiscus L.), yarrow (Achillea millefolium L.) and myrtle (Myrtus communis L.) grow in the mild climate of Eurasia and are receiving particular attention for their medicinal and aromatic value. Two distinct concentrations of selected essential oils (EOs) were tested, dispersed in organogels based on 3 % (m/V) Carbopol 974P NF in PEG 400. Prior to formulation development, selected EOs were characterized by mass spectroscopy to identify the dominating bioactives. A total of 32 compounds were quantified for lentisk leaf EO, 42 for yarrow EO, and 26 for myrtle leaf EO. The best formulations were found for the highest concentration of EOs (4 %, m/m) into Carbopol/PEG400 organogels. Organogels were produced by high-speed homogenization followed by microwave heating and showed acceptable pH between 5.8 and 6.0, being compatible with the skin, with an electrical conductivity highest for yarrow EO-based organogel (YIG4%, ∼64 mV), and the lowest for myrtle EO-based organogel (MIG4%, ∼48 mV). Texture analysis showed the highest adhesion properties for lentisk EO-based organogel (LIG4%), attributed to its higher viscosity recorded for the two tested temperatures (25 °C vs 37 °C). Rheological analysis confirmed that organogels have pseudoplastic behaviour. The antibacterial activity was compared between plain EOs and EOs-based organogels, with the yarrow EO and its organogel (YIG4%) depicting the highest activity against both tested bacterial strains (S. aureus and C. acnes). On the other hand, MIG4% contributed for the highest cell proliferation rate tested in a mouse embryo fibroblast cell line (3T3-L1), whereas with LIG4% the lowest viability value was recorded.

Comparing Metabolomic and Essential Oil Fingerprints of Citrus australasica F. Muell (Finger Lime) Varieties and Their In Vitro Antioxidant Activity.

Comparative chemical analyses among peel and pulp essential oils (EOs) and methanolic extracts of four Citrus australasica varieties (Red, Collette, Pink Ice, and Yellow Sunshine), and the hybrid Faustrime, were performed using GC-MS and UHPLC-DAD-HR-Orbitrap/ESI-MS. Peel and pulp extracts were also analysed for their in vitro antioxidant activity on a Balb/3T3 clone A31 mouse embryo fibroblast cell line. The results of peel and pulp EOs were mainly characterised by monoterpenes and sesquiterpenes, respectively. All peels displayed a higher total phenol content (TPC) than pulps, and consequently a greater antioxidant activity. Collette peels and Pink Ice pulps showed the highest amount of identified flavonoids (e.g., luteolin, isosakuranetin, and poncirin derivatives). Collette and Red peels were rich in anthocyanins (delphinidin and petunidin glycosides), exhibiting the maximum protective activity against induced oxidative damage. In conclusion, finger lime fruits are good sources of health-promoting phytocomplexes, with the Red, Collette, and Pink Ice varieties being the most promising.

Incorporation of Mycobacteriophage Fulbright into Polycaprolactone Electrospun Nanofiber Wound Dressing.

The Genus Mycobacterium includes pathogens known to cause disease in mammals such as tuberculosis (Mycobacterium tuberculosis) and skin infections (M. abscessus). M. smegmatis is a model bacterium that can cause opportunistic infections in human tissues and, rarely, a respiratory disease. Due to the emergence of multidrug-resistant bacteria, phage therapy is potentially an alternative way of treating these bacterial infections. As bacteriophages are specific to their bacterial host, it ensures that the normal flora is unharmed. Fulbright is a mycobacteriophage that infects the host bacteria M. smegmatis. The main goal of this study is to incorporate Mycobacteriophage Fulbright into a polycaprolactone (PCL) nanofiber and test its antimicrobial effect against the host bacteria, M. smegmatis. Stability tests conducted over 7 days showed that the phage titer does not decrease when in contact with PCL, making it a promising vehicle for phage delivery. Antimicrobial assays showed that PCL_Fulbright effectively reduces bacterial concentration after 24 h of contact. In addition, when stored at −20 °C, the phage remains viable for up to eleven months in the fiber. Fulbright addition on the nanofibrous mats resulted in an increase in water uptake and decrease in the mechanical properties (strength and Young’s modulus) of the membranes, indicating that the presence of phage Fulbright can greatly enhance the physical and mechanical properties of the PCL. Cytotoxicity assays showed that PCL_Fulbright is not cytotoxic to Balbc/3T3 mouse embryo fibroblast cell lines; thus, phage-incorporated PCL is a promising alternative to antibiotics in treating skin infections.

Development of Chitosan Silver Nanocomposites: Its Characteristic Study and Toxicity Effect against 3T3-L1 Cell Line

Nanocomposite from the natural source is opened a wide area for the researchers to find a natural remedy to replace the chemicals or harmful products in all the fields of agricultural, food and medical fields. Here the biopolymer (chitosan) was extracted from the two white rot fungi of Pleurotus floridanus and Pleurotus djamor, and biologically synthesized with 1mM AgNO3 solution. Synthesized chitosan nanocomposite was characterized with UV-Visible study, FTIR, FESEM, XRD, EDAS for the confirmation based upon the peaks, functional group, crystalline nature, size, morphology and the percentage of elements respectively. Toxicity study was carried out using 3T3 L1 (Mouse embryo fibroblast Cell Line) normal Cell Line to find out the cytotoxicity effect of the chitosan nanocomposite and found that the nanocomposites were non toxic to the Cell Line.

Photocatalytic performance and cytotoxic activity of green-synthesized cobalt ferrite nanoparticles

In this study, cobalt ferrite nanoparticles (CoFe2O4 NPs) were prepared using aqueous solution of torajabin followed by calcination at temperatures of 600, 700, and 800 °C. Synthesized nanoparticles of size 20–60 nm were characterized using PXRD, UV–Vis, FESEM, EDX, and Raman analysis. VSM analysis affirmed ferrimagnetic behavior. The photocatalytic degradation activity of CoFe2O4 NPs was assessed on Acid Orange 7 (AO7) dye under simulated sunlight exposure where high level of photodegradation was displayed by CF-600 NPs, due to their small particle size. The cytotoxic activity of CoFe2O4 NPs was studied on breast cancer cell (MCF7), colon cancer cell (CaCo2), and mouse embryo fibroblast cell (NIH-3T3) lines using MTT assay; nanoparticles were found to be nontoxic against cancer and normal cell lines. Thus, synthesized CoFe2O4 NPs could be an ideal choice for medical applications like drug delivery, and non-medical usages such as photodegradation of pollutants.

Novel N-(2-Methoxydibenzofuran-3-yl)-2-aryloxyacetamide Derivatives: Synthesis and Biological Investigation

Background: Dibenzofuran ring is a typical heterocyle that is found in many natural sources and its derivatives exhibit a wide scale of biological applications similar to its analog ring systems; furan and benzofuran. Materials and Methods: Novel N-(2-methoxydibenzofuran-3-yl)-2-aryloxyacetamide derivatives (2a-l) were synthesized and evaluated for their cytotoxic activity against A549 lung cancer and NIH/3T3 mouse embryofibroblast cell lines. The inhibition percentages of cathepsin D, L, acetylcholinesterase (AChE) and butrylcholinesterase (BuChE) enzymes provoked by the compounds were also determined. Results and Discussion: Most of the compounds exhibited significant cytotoxicity whose IC50 values were identified lower than the tested lowest concentration (<3.90 μg/mL). Compound 2i against cathepsin D and compound 2k against cathepsin L displayed the highest inhibitory activity. Regrettably, the compounds demonstrated very weak AChE and BuChE inhibition. Conclusion: Compounds 2b, 2c, 2e, 2i and 2k exhibited the highest antiproliferative activity against A549 cell lines with selective profile. However, they did not display satisfying results on tested enzymes.

Abstract PR-003: Radiosensitization by targeting the NPM1/RAD51 axis

Abstract NPM1 is a chaperone involved in base excision, translesion synthesis and homologous recombination (HR). In the presence of DNA DSBs pT199 NPM1 co-localizes with H2AX foci, a consequence of binding to K63-linked ubiquitinated histones that surround DSBs via its pT199 domain comprised of an acidic tract and adjacent ubiquitin interacting motif-like domain (1-3). SUMOlyated K263 NPM1 binds to and promotes Rad51 loading onto resected 3-prime DNA at the DSB. Failure to SUMOlyate K263NPM or inhibition of NPM1 reduces Rad51 filament formation, thus inhibiting HR (4). A major effort is underway to develop DNA damage response inhibitors. The novel chemical entity YTR107 is a 5-((N-benzyl-1H-indol-3-yl) methylene) pyrimidine-2, 4,6(1H,3H,5H) trione developed using forward chemical genetics and structure/function screening (5). Colony formation assays revealed that YTR107 radiosensitized 7 NSCLC lines (dose modifying factors of 1.5). Radiosensitization was NPM1-dependent as YTR107 did not radiosensitize an NPM1 null mouse embryo fibroblast cell line (3). Here we show that YTR107 inhibits radiation-induced NPM1 binding to Rad51, Rad 51 foci formation and repair of DNA DSBs.Tumor initiating cells (TICs) were radiosensitized by YTR107. YTR107 inhibited tumor growth (P = 0.027, 2-way ANOVA) and increased the survival of A549 tumor-bearing mice, determined 70 days after the last treatment. Survival of tumor-bearing mice was 60% for mice treated with YTR107and 2.2 Gy(q.d. x 7) vs 20% for mice treated with 2.2 Gy alone (q.d.x 7). Cox Proportional log rank hazard ratio for irradiated vs unirradiated tumor-bearing mice was 0.24 (P = 0.0034) while the log rank hazard ratio for YTR107 plus irradiation vs unirradiated (± YTR107) tumor-bearing mice was 0.17 (P = 0.0023). Thus, these data provide support for development of YTR107 as a therapeutic radiation sensitizer. Supported in part by R44CA228756. 1.Koike A, Nishikawa H, Wu W, Okada Y, Venkitaraman AR, Ohta T.Cancer Res 2010;70:6746-56 2. Sekhar KR, Benamar M, Venkateswaran A, Sasi S, Penthala NR, Crooks PA, et al. Int J Radiat Oncol Biol Phys 2014;89:1106-14 3. Sekhar KR, Reddy YT, Reddy PN, Crooks PA, Venkateswaran A, McDonald WH, et al. Clin Cancer Res 2011;17:6490-9 4. Xu R, Yu S, Zhu D, Huang X, Xu Y, Lao Y, et al. Nat Commun 2019;10:3812 5. Reddy YT, Sekhar KR, Sasi N, Reddy PN, Freeman ML, Crooks PA. Bioorg Med Chem Lett 2010;20:600-2 Citation Format: Michael L. Freeman, Geri Traver, Konjeti R. Sekhar, Peter A. Crooks. Radiosensitization by targeting the NPM1/RAD51 axis [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PR-003.

The mTORC1-mediated activation of ATF4 promotes protein and glutathione synthesis downstream of growth signals.

The mechanistic target of rapamycin complex 1 (mTORC1) stimulates a coordinated anabolic program in response to growth-promoting signals. Paradoxically, recent studies indicate that mTORC1 can activate the transcription factor ATF4 through mechanisms distinct from its canonical induction by the integrated stress response (ISR). However, its broader roles as a downstream target of mTORC1 are unknown. Therefore, we directly compared ATF4-dependent transcriptional changes induced upon insulin-stimulated mTORC1 signaling to those activated by the ISR. In multiple mouse embryo fibroblast and human cancer cell lines, the mTORC1-ATF4 pathway stimulated expression of only a subset of the ATF4 target genes induced by the ISR, including genes involved in amino acid uptake, synthesis, and tRNA charging. We demonstrate that ATF4 is a metabolic effector of mTORC1 involved in both its established role in promoting protein synthesis and in a previously unappreciated function for mTORC1 in stimulating cellular cystine uptake and glutathione synthesis.

3,4,5-Trisubstituted-1,2,4-triazole Derivatives as Antiproliferative Agents: Synthesis, In vitro Evaluation and Molecular Modelling

Background: Cancer is the name given to various diseases that are mainly uncontrolled, related to cell growth and can affect various organs. Among them, lung cancer is the one, which, in its earliest stages, is difficult to diagnose, and it is asymptomatic until the disease progresses. Triazole ring is an important heterocyclic ring known with various pharmacological activities. Objective: It is aimed to synthesize and characterize novel 1,2,4-triazole derivatives and screen them for in vitro antiproliferative activity and binding analysis through docking studies. Method: In this study, we have synthesized new 2-[[5-[(4-aminophenoxy)methyl]-4-phenyl-4H- 1,2,4-triazol-3-yl]thio]-N-(substituted aryl)acetamide (5a-h) derivatives and investigated their anticancer activities against human lung cancer (A549) and mouse embryo fibroblast cell lines (NIH/3T3) by MTT, flow cytometric, caspase-3 and matrix metalloproteinase-9 (MMP-9) inhibition assays. Results: Compounds 5f, 5g and 5h showed the highest cytotoxicity and caused significant apoptosis. These compounds inhibited MMP-9, slightly whereas they did not effect caspase-3. Conclusion: 5f namely, N-(5-acetyl-4-methylthiazol-2-yl)-2-((5-((4-aminophenoxy)methyl)-4- phenyl-4H-1,2,4-triazol-3-yl)thio)acetamide exhibited as the most active compound with selective cytotoxicity and the highest MMP-9 inhibition. Besides, molecular modelling assessment was signified that antiproliferative activity of the compounds 5f, 5g and 5h was through a slight MMP-9 inhibition pathway.

Abstract 6164: Small-molecule targeting of the NPM1/Rad51 repair pathway for the radiosensitization of cancer

Abstract Nucleophosmin1 (NPM1) is rapidly recruited to sites of DNA double-strand breaks and is required for radiation-induced Rad51 foci formation. Inhibition of NPM1 or Rad51 is a radiosensitizing event. YTR107 is a small molecule, identified in a forward chemical genetics functional structure/activity screening, that binds to the amino terminus of NPM1 and prevents NPM1 recruitment to damaged chromatin. In silico docking studies of YTR107 and the crystal structure of the NPM1 N-terminal oligomerization domain revealed that YTR107 docked to monomeric NPM1 in a pocket crucial for forming an interface between neighboring subunits of the NPM1 pentamer. YTR107 inhibited radiation-induced Rad51 foci formation and repair of DNA DSBs, as measured by a neutral comet assay. Colony formation assays revealed that YTR107 radiosensitized 7 NSCLC lines, HT29 colorectal cancer cells, D54 glioblastoma cells, PANC1 pancreatic cancer cells, and 2 breast cancer cell lines (dose modifying factors of > 1.5; cells irradiated with 300 kVp/10mA X-rays @ 2 Gy/min). Radiosensitization was found to be NPM1-dependent as YTR107 did not radiosensitize an NPM1 null mouse embryo fibroblast cell line. YTR107 produced a 4.7 fold increase in HT29 xenograft growth delay (time required for the tumor to increase 4-fold in volume (p = 0.001) following 10 mg/kg YTR107 administered 30 min prior to administering 3 Gy to the tumor for 7 consecutive days). YTR107 increased the survival of A549 tumor-bearing mice determined 70 days after the last treatment. Survival of tumor-bearing mice was 60% for mice treated with YTR107 and 3 Gy vs 20% for mice treated with 3 Gy alone, p = 0.0012 (0 or 20 mg/kg YTR107 administered 1 hr prior to 0 or 2.2 Gy being administered to the tumor for 7 consecutive days). Female C57BL/6J mice were administered 0 or 10 mg/kg i.p. YTR107 for 5 consecutive days. Thirty five days after injection mice were euthanized and subjected to necropsy by a veterinarian trained in veterinary pathology. Gross and histological examination of liver, lung, thymus, heart, spleen, cerebellum, pancreas, small intestine, kidney, and adrenal gland did not reveal evidence of toxicity. Both a complete blood count and a white blood cell differential count were performed. No significant differences were noted. Mice were subjected to five daily injections of 30 mg/kg YTR107. This dose level was also tolerated without overt toxicity after 35 days. Mice were injected with 0 or 30 mg/kg YTR107 (i.p.) and 30 min later administered 0 or 16-Gy to the thorax. Pulmonary fibrotic lesions were quantified using respiratory-gated micro CT 136 days later. YTR107 did not increase the severity of radiation-induced lung fibrosis and did not affect body weights or life span of unirradiated or irradiated mice (p > 0.05) over the 136-day study. Thus, this study provides support for development of YTR107 as a therapeutic radiation sensitizer and was supported in part by R44CA228756-02 Citation Format: Konjeti R. Sekhar, Geri Traver, Peter A. Crooks, Michael L. Freeman. Small-molecule targeting of the NPM1/Rad51 repair pathway for the radiosensitization of cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6164.

A versatile cell-friendly approach to produce PLA-based 3D micro-macro-porous blends for tissue engineering scaffolds

In this study we instituted an innovative, cost-effective, green and versatile methodology to produce a series of PLA-based open-pore porous blends with high porosity and interconnectivity as well physico-mechanical properties suitable for tissue engineering application. Parent poly-l-lactic acid (PLA) blend was prepared by melt-blending using crosslinked sodium polyacrylate particles as a porogen, commonly used as superabsorbent polymer (SAP). The obtained biphasic systems showed a regular distribution of SAP particles with diameters up to about 50 µm and, most importantly, retained their superabsorbent ability within the thermoplastic PLA based matrices that facilitated swelling followed by leaching out from PLA based matrices in aqueous environment generating very high porosity. Very importantly, versatility of this developed methodology was judged by accommodating different polymers, such as, poly (3-hydroxybutyrate) (PHB) poly (ɛ-caprolactone) (PCL) or poly (ethylene glycol) (PEG) or wood-cellulose microfiber (SP) to generate monophasic, biphasic, plasticized or reinforced blends, respectively, under identical benign condition. These blends were analyzed morphologically, thermally and mechanically to evaluate the degree of miscibility, thermal stability and mechanical property to apply as scaffolds in tissue engineering. Finally, all these scaffolds allowed good cell adhesion and proliferation during culture of mouse embryo fibroblasts cell line. Hence, this methodology of producing PLA-based polymeric system stunned with processability to accommodate other biocompatible polymers allows selectively modifying biomaterial properties for target application and appears very promising platform for several applications, particularly for scaffold production in tissue engineering.

A Versatile Cell-Friendly Approach to Produce PLA-Based 3D Micro-Macro-Porous Blends for Tissue Engineering Scaffolds

In this study we instituted an innovative, cost-effective, green and versatile methodology to produce a series of PLA-based open-pore porous blends with high porosity and interconnectivity as well physico-mechanical properties suitable for tissue engineering application. Parent poly-L-lactic acid (PLA) blend was prepared by melt-blending using crosslinked sodium polyacrylate particles as a porogen, commonly used as superabsorbent polymer (SAP). The obtained biphasic systems showed a regular distribution of SAP particles with diameters up to about 50 μm and, most importantly, retained their superabsorbent ability within the thermoplastic PLA based matrices that facilitated swelling followed by leaching out from PLA based matrices in aqueous environment generating very high porosity. Very importantly, versatility of this developed methodology was judged by accommodating different polymers, such as, poly (3-hydroxybutyrate) (PHB) poly (ɛ-caprolactone) (PCL) or poly (ethylene glycol) (PEG) or wood-cellulose microfiber (SP) to generate monophasic, biphasic, plasticized or reinforced blends, respectively, under identical benign condition. These blends were analyzed morphologically, thermally and mechanically to evaluate the degree of miscibility, thermal stability and mechanical property to apply as scaffolds in tissue engineering. Finally, all these scaffolds allowed good cell adhesion and proliferation during culture of mouse embryo fibroblasts cell line. Hence, this methodology of producing PLA-based polymeric system stunned with processability to accommodate other biocompatible polymers allows selectively modifying biomaterial properties for target application and appears very promising platform for several applications, particularly for scaffold production in tissue engineering.

PEG-PCL Nanocapsules Containing Curcumin: Validation of HPLC Method for Analyzing Drug-loading Efficiency, Stability Testing and Cytotoxicity on NIH-3T3 Cell Line

Curcumin (CUR) shows potential use for treating cancer. However, CUR has low solubility and reduced bioavailability, which limit its clinical effect. Therefore, the development of nanocarriers can overcome these problems and can ensure the desired pharmacological effect. In addition, it is mandatory to prove the quality, the efficacy, and the safety for a novel nanomedicine to be approved. In that sense, this paper aimed (a) to prepare CUR-loaded polyethylene glycol-poly(ε-caprolactone) nanocapsules; (b) to validate an analytical method by high performance liquid chromatography (HPLC) for quantifying CUR in these nanoformulations; (c) to evaluate the physicochemical stability of these formulations; and to investigate their cytotoxicity on NIH-3T3 mouse fibroblast cells. The HPLC method was specific to CUR in the loaded nanocapsules, linear (r = 0.9994) in a range of 10.0 to 90.0 µg.mL-1 with limits of detection and quantification of 0.160 and 0.480 µg.mL-1, respectively. Precision was demonstrated by a relative standard deviation lower than 5%. Suitable accuracy (102.37 ± 0.92%) was obtained. Values of pH, particle size, polydispersity index, and zeta potential presented no statistical difference (p > 0.05) for CUR-loaded nanoparticles. No cytotoxicity was observed against NIH-3T3 mouse embryo fibroblast cell line using both the tetrazolium salt and sulforhodamine B assays. In conclusion, a simple and inexpensive HPLC method was validated for the CUR quantification in the suspensions of nanocapsules. The obtained polymeric nanocapsules containing CUR showed suitable results for all the performed assays and can be further investigated as a feasible novel approach for cancer treatment.

Preparation, characterization, and biological evaluation of retinyl palmitate and Dead Sea water loaded nanoemulsions toward topical treatment of skin diseases

Millions of people suffer from different types of skin diseases worldwide. In the last decade, the development of nanocarriers has been the focus of the pharmaceutical and cosmetic industries to enhance the performance of their products, and to meet consumers’ demands. Several delivery systems have been developed to improve the efficiency and minimize possible side effects. In this study, retinyl palmitate and Dead Sea water loaded nanoemulsions were developed as carriers to treat skin conditions such as photoaging, psoriasis, or atopic dermatitis. Toxicity profiles were carried out by means of viability, cell membrane asymmetry study, evaluation of oxidative stress induction (reactive oxygen species), and inflammation via cytokines production with a human keratinocyte cell line (HaCaT) and a mouse embryo fibroblasts cell line (BALB/3T3). Results showed that loaded nanoemulsions were found to be non-cytotoxic under the conditions of the study. Furthermore, no oxidative stress induction was observed. Likewise, an efficacy test of these loaded nanoemulsions was also tested on human skin organ cultures, before and after ultraviolet B light treatment. Viability and caspase-3 production assessment, in response to the exposure of skin explants to the loaded nanoemulsions, indicated non-toxic effects on human skin in culture, both with and without ultraviolet B irradiation. Further the ability of loaded nanoemulsions to protect the skin against ultraviolet B damage was assessed on skin explants reducing significantly the apoptotic activation after ultraviolet B irradiation. Our promising results indicate that the developed loaded nanoemulsions may represent a topical drug delivery system to be used as an alternative treatment for recurrent skin diseases.

Two metabolites isolated from endophytic fungus Coniochaeta sp. F-8 in Ageratina adenophora exhibit antioxidative activity and cytotoxicity

Ageratina adenophora is an invasive plant of global importance and has a broad distribution in the Western Himalayas. Endophtytic fungus Coniochaeta sp.F-8 was found in Ageratina adenophora. In this paper, we aim to investigate the antioxidative activity and cytotoxicity of the metabolites from Coniochaeta sp.F-8. Consequently, two compounds Phomoxanthone A and Penialidin A were isolated from the endophytic fungus of Ageratina adenophora for the first time. The structures of compounds were identified by IR, NMR and ESI-MS methods. Moreover, both of those compounds showed considerable antioxidative activity in vitro and resulted cytotoxicity in mouse embryo fibroblasts cell line Balb/c3T3. The present study provides a theoretical foundation for the development and utilization of endophytic fungi in Ageratina adenophora as a medicinal substance.

Some Thiazole Derivatives Combined with Different Heterocycles: Cytotoxicity Evaluation and Apoptosis Inducing Studies.

Thiazole ring is an outstanding structure found in many biologically active compounds and clinically available drugs. Because of synthesis simplicity of its derivatives and having a wide range of biological aspects along with high effectiveness, new thiazole derivatives have been studied by medicinal chemists since many years. Some thiazole compounds combined with different heterocyclic rings were acquired in this study. Novel 5-(4-substituted benzylidene)-2-[(4,5-dimethylthiazol-2-yl)amino]thiazol-4(5H)-one derivatives (4a-g) and 2-(heteroaryl-2/3/5-yl)thio-N-(4,5-dimethylthiazol-2-yl)acetamide (4h-p) derivatives were synthesized starting from 2-amino-4,5-dimethylthiazole. The obtained compounds were evaluated to determine their antiproliferative activity. Final compounds (4a-g) were obtained with a ring closure reaction; other final compounds (4h-p) were acquired via the reaction of mercapto heterocycles with Hantzsch thiazole synthesis intermediate. To evaluate antiproliferative activity of them, the compounds were tested on A549 adenocarcinomic human alveolar basal epithelial cells line, C6 rat glioma cell line and NIH/3T3 mouse embryo fibroblast cell line according to the conventional MTT method. Besides, the selected compounds were studied to find out which pathway cell deaths caused via Annexin V/PI staining. Compounds 4f, 4j and 4p exhibited the highest cytotoxicity on A549 with a non-toxic profile. Also, 4f, 4h, 4j and 4p were determined as the most antiproliferative compounds on C6 cell line. Furthermore, compound 4p induced apoptosis of A549 cell with a level of 23.5% and compound 4h induced C6 cell with a level of 37.5%. Considering the structure of the compounds, although thiazolidine containing compounds 4a-g exhibited higher activity in general, compounds 4p containing 5-chlorobenzothiazole moiety showed the highest cytotoxicity. It could be expressed with the conspicuous antitumor efficiency of benzothiazole ring.

Strawberry extract attenuates oxidative stress in 3T3-L1 cells

BACKGROUND: High levels of reactive oxygen species (ROS) within the adipose tissue promote a disturbed redox balance and influence its function, impairing adipogenesis, inducing insulin resistance and stimulating adipocyte hypertrophy. Supplementation with antioxidant rich foods can reverse some of these effects. Strawberry is well known as a good source of phytochemicals; however, whether strawberry suppresses increased oxidative stress in 3T3-L1 cells remain unclear. OBJECTIVE: The purpose of the present work was to determine the antioxidant potential of a strawberry extract in 3T3-L1 mouse embryo fibroblast cell line. METHODS: 3T3-L1 pre-adipocytes were induced to differentiate into adipocytes in the presence or absence of different concentrations of the strawberry extract. At the end of the differentiation period, intracellular ROS production, thiobarbituric acid reactive substances (TBARS) content, as well as superoxide dismutase (SOD) and catalase (CAT) activities and gene expressions were evaluated. RESULTS: In this study, we confirmed that strawberry extract markedly inhibited increased-oxidative stress in 3T3L1 cells by suppressing intracellular ROS production and decreasing TBARS content. Likewise, SOD and CAT activities and gene expressions were increased. CONCLUSIONS: This paper provides evidence that strawberry extract is able to scavenging free radicals and activate endogenous defense systems, highlighting its potential capacity to modulate obesity induced- inflammatory states.

Synthesis of New Benzothiazole Acylhydrazones as Anticancer Agents.

During the last five decades, a large number of BT (Benzothiazole) derivatives formed one of the eligible structures in medicinal chemistry as anticancer agents. Most of the studies reveal that various substitutions at specific positions on BT scaffold modulate the antitumor property. The potential of BTs encouraged us to synthesize a number of new 2-((5-substitutedbenzothiazol-2-yl)thio)-N’-(2-(4-(substitutedphenyl)ethylidene)acetohydrazide derivatives and investigate their probable anticancer activity. 4-Substitued benzaldehyde derivatives (1a–1e) were afforded by the reaction of appropriate secondary amine and 4-fluorobenzaldehyde in DMF. Equimolar quantitates of 5-substitutedbenzothiazole-2-thiol, ethyl chloroacetate and K2CO3 were refluxed in acetone to obtain 2-((5-substitutedbenzothiazol-2-yl)thio)acetate derivatives (2a,2b), which reacted with excess of hydrazine hydrate to get 2-((5-substitutebenzothiazol-2-yl)thio)acetohydrazides (3a,3b). In the last step, 2-((5-substitutedbenzothiazol-2-yl)thio)-N’-(4-substitutedbenzylidene)acetohydrazide derivatives (4a–4j) were synthesized by the reaction of 1a–1e and 3a–3b in EtOH. The anticancer activity of target compounds was evaluated in three steps. First, an MTT test (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) was performed to observe cytotoxic activity of the compounds against carcinogenic C6 (Rat brain glioma cell line), A549 (Human lung adenocarcinoma epithelial cell line), MCF-7 (Human breast adenocarcinoma cell line), and HT-29 (Human colorectal adenocarcinoma cell line) cancer cell lines. Healthy NIH3T3 (Mouse embryo fibroblast cell line) cells were also subjected to MTT assay to determine selectivity of the compounds towards carcinogenic cell lines. Secondly, inhibitory effects of selected compounds 4d, 4e, and 4h on DNA synthesis of C6 cells were investigated. Finally, flow cytometric analysis were performed to identify the death pathway of the carcinogenic cells.

Diclofop-methyl: A phenoxy propionate herbicide with multiple toxic effects in mouse embyro fibroblast (NIH/3T3) cell line

Diclofop-methyl is a selective post-emergence graminicide from the phenoxy propionate group of herbicides to be developed for control of wild oats, millets, and other annual grass weeds. Diclofop-methyl usage is limited in various grass weed species due to its toxic effect and exposure risks. However, total annual usage of is approximately 750.000 pounds in United States, and more in Asia. Therefore, we aimed to investigate diclofopmethyl’s toxic potentials in vitro and the following assays were used; MTT assay for cytotoxicity, comet assay for genotoxicity, generation of reactive oxygen species (ROS), malondialdehyde (MDA) and glutathione (GSH) for the potential of oxidative damage in mouse embryo fibroblast (NIH/3T3) cell line. Diclofop-methyl was observed to reduce the cell viability in a concentration manner and the half maximal inhibitory concentration (IC50) value was 301.7 μM. Diclofop-methyl caused DNA damage and oxidative stress at the concentrations between 12.5-400 μM. Tail intensities were at the range of 1.24- 58.21% with increasing concentrations, which are approximately ≤ 1.63-fold of the negative control. Also, MDA levels were increased ≥ ³11.4-fold of the negative control that denotes lipid peroxidation was induced. However, there was no significant increment in the ROS and GSH levels at all concentrations. In view of the fact that ROS has not been detected, despite its level of MDA proffers, the idea that oxidative damage may have been caused by other mechanisms. Our results indicate that diclofopmethyl was cytotoxic, genotoxic and might have oxidative damage potential in vitro conditions.

Cytotoxic effect of green tea leaf extract on tumor cell line

The study was conducted to evaluate antitumor effects of green tea (Camellia sinensis) extracts (aqueous and methanolic) on Rhabdomyosarcoma; cell line and a normal cell line; mouse embryo fibroblast; Chemical detections of green tea extracts revealed that the aqueous and methanolic extracts were positive for flavonoids, alkaloids, phenol and glycosides. The percentage growth inhibition of five plant concentrations (50, 100, 250, 500 and 1000 µg/ml) were assessed in vitro using tumor cell lines Rhabdomyosarcoma and normal cell line mouse embryonic fibroblasts. The results revealed that the five concentrations of the plant extracts showed anti-tumor properties in a concentration-dependent manner, and the methanolic extract recorded better values of percentage growth inhibition than the aqueous extract in Rhabdomyosarcoma cell lines, while, less percentage growth inhibition values were recorded in the mouse embryo fibroblast cell line.