Search results for: in vitro metabolites

Authors: Nima Samie, Sekaran Muniandy, Zahurin Mohamed, M. S. Kanthimathi

Abstract:

Although number of indole containing compounds have been reported to have anticancer properties in vitro but only a few of them show potential as anticancer compounds in vivo. The current study was to evaluate the mechanism of cytotoxicity of selected indolic compound in vivo and in vitro. In this context, we determined the potency of the compound in the induction of apoptosis, cell cycle arrest, and cytoskeleton rearrangement. HT-29, WiDr, CCD-18Co, human monocyte/macrophage CRL-9855, and B lymphocyte CCL-156 cell lines were used to determine the IC50 of the compound using the MTT assay. Analysis of apoptosis was carried out using immunofluorescence, acridine orange/ propidium iodide double staining, Annexin-V-FITC assay, evaluation of the translocation of NF-kB, oxygen radical antioxidant capacity, quenching of reactive oxygen species content, measurement of LDH release, caspase-3/-7, -8 and -9 assays and western blotting. The cell cycle arrest was examined using flowcytometry and gene expression was assessed using qPCR array. Results displayed a potent suppressive effect on HT-29 and WiDr after 24 h of treatment with IC50 value of 2.52±0.34 µg/ml and 2.13±0.65 µg/ml respectively. This cytotoxic effect on normal, monocyte/macrophage and B-cells was insignificant. Dipping in the mitochondrial membrane potential and increased release of cytochrome c from the mitochondria indicated induction of the intrinsic apoptosis pathway by the compound. Activation of this pathway was further evidenced by significant activation of caspase-9 and 3/7. The compound was also shown to activate the extrinsic pathways of apoptosis via activation of caspase-8 which is linked to the suppression of NF-kB translocation to the nucleus. Cell cycle arrest in the G1 phase and up-regulation of glutathione reductase, based on excessive ROS production were also observed. These findings were further investigated for inhibitory efficiency of the compound on colonic aberrant crypt foci in male rats. Rats were divided in to 5 groups: vehicle, cancer control, positive control groups and the groups treated with 25 and 50 mg/kg of compounds for 10 weeks. Administration of compound suppressed total colonic ACF formation up to 73.4%. The results also showed that treatment with the compound significantly reduced the level of malondialdehyde while increasing superoxide dismutase and catalase activities. Furthermore, the down-regulation of PCNA and Bcl2 and the up-regulation of Bax was confirmed by immunohistochemical staining. The outcome of this study suggest sthat the indolic compound is a potent anti-cancer agent against colon cancer and can be further evaluated by animal trial.

Keywords: indolic compound, chemoprevention, crypt, azoxymethane, colon cancer

Procedia PDF Downloads 334

Authors: Thaís T. Valério Caetano, João Máximo De Siqueira, Carlos Alexandre Carollo, Arthur Ladeira Macedo, Vanessa C. Stein

Abstract:

Nicotiana glauca, commonly known as wild tobacco or tobacco bush, belongs to the Solanaceae family. It is native to South America but has become naturalized in various regions, including Australia, California, Africa, and the Mediterranean. N. glauca is listed in the Global Invasive Species Database (GISD) and the Invasive Species Compendium (CABI). It is known for producing pyridine alkaloids, including anabasine, which is highly toxic. Anabasine is predominantly found in the leaves and can cause severe health issues such as neuromuscular blockade, respiratory arrest, and cardiovascular problems when ingested. Mistaken identity with edible plants like spinach has resulted in food poisoning cases in Israel and Brazil. Anabasine, a minor alkaloid constituent of tobacco, may contribute to tobacco addiction by mimicking or enhancing the effects of nicotine. Therefore, it is essential to investigate the production pattern of anabasine and its relationship to the developmental stages of the plant. This study aimed to establish the relationship between the phenological plant age, cultivation place, and the increase in anabasine concentration, which can lead to human intoxication cases. In this study, N. glauca plants were collected from three different rural areas in Brazil for a year to examine leaves at various stages of development. Samples were also obtained from cultivated plants in Marilândia, Minas Gerais, Brazil, as well as from Divinópolis, Minas Gerais, Brazil, and Arraial do Cabo, Rio de Janeiro, Brazil. In vitro cultivated plants on MS medium were included in the study. The collected leaves were dried, powdered, and stored. Alkaloid extraction was performed using a methanol and water mixture, followed by liquid-liquid extraction with chloroform. The anabasine content was determined using HPLC-DAD analysis with nicotine as a standard. The results indicated that anabasine production increases with the plant's development, peaking in adult leaves during the reproduction phase and declining afterward. In vitro, plants showed similar anabasine production to young leaves. The successful adaptation of N. glauca in new environments poses a global problem, and the correlation between anabasine production and the plant's developmental stages has been understudied. The presence of substances produced by the plant can pose a risk to other species, especially when mistaken for edible plants. The findings from this study shed light on the pattern of anabasine production and its association with plant development, contributing to a better understanding of the potential risks associated with N. glauca and the importance of accurate identification.

Keywords: nicotiana glauca graham, global invasive species database, alkaloids, toxic

Procedia PDF Downloads 61

Authors: Yang Yang, Luciana Magalhães Rebelo Alencar, Martha Sahylí Ortega Pijeira, Beatriz da Silva Batista, Alefe Roger Silva França, Erick Rafael Dias Rates, Ruana Cardoso Lima, Sara Gemini-Piperni, Ralph Santos-Oliveira

Abstract:

Radium-²²³ dichloride ([²²³Rₐ]RₐCl₂) is an alpha particle-emitting radiopharmaceutical currently approved for the treatment of patients with castration-resistant prostate cancer, symptomatic bone metastases, and no known visceral metastatic disease. [²²³Rₐ]RₐCl₂ is bone-seeking calcium mimetic that bonds into the newly formed bone stroma, especially osteoblastic or sclerotic metastases, killing the tumor cells by inducing DNA breaks in a potent and localized manner. Nonetheless, the successful therapy of osteosarcoma as primary bone tumors is still a challenge. Nanomicelles are colloidal nanosystems widely used in drug development to improve blood circulation time, bioavailability, and specificity of therapeutic agents, among other applications. In addition, the enhanced permeability and retention effect of the nanosystems, and the renal excretion of the nanomicelles reported in most cases so far, are very attractive to achieve selective and increased accumulation in tumor site as well as to increase the safety of [²²³Rₐ]RₐCl₂ in the clinical routine. In the present work, [²²³Rₐ]RₐCl₂ nanomicelles were produced, characterized, in vitro evaluated, and compared with pure [²²³Rₐ]RₐCl2 solution using SAOS2 osteosarcoma cells. The [²²³Rₐ]RₐCl₂ nanomicelles were prepared using the amphiphilic copolymer Pluronic F127. The dynamic light scattering analysis of freshly produced [²²³Rₐ]RₐCl₂ nanomicelles demonstrated a mean size of 129.4 nm with a polydispersity index (PDI) of 0.303. After one week stored in the refrigerator, the mean size of the [²²³Rₐ]RₐCl₂ nanomicelles increased to 169.4 with a PDI of 0.381. Atomic force microscopy analysis of [223Rₐ]RₐCl₂ nanomicelles exhibited spherical structures whose heights reach 1 µm, suggesting the filling of 127-Pluronic nanomicelles with [²²³Rₐ]RₐCl₂. The viability assay with [²²³Rₐ]RₐCl₂ nanomicelles displayed a dose-dependent response as it was observed using pure [²²³Rₐ]RₐCl2. However, at the same dose, [²²³Rₐ]RₐCl₂ nanomicelles were 20% higher efficient in killing SAOS2 cells when compared with pure [²²³Rₐ]RₐCl₂. These findings demonstrated the effectiveness of the nanosystem validating the application of nanotechnology in targeted alpha therapy with [²²³Ra]RₐCl₂. In addition, the [²²³Rₐ]RaCl₂nanomicelles may be decorated and incorporated with a great variety of agents and compounds (e.g., monoclonal antibodies, aptamers, peptides) to overcome the limited use of [²²³Ra]RₐCl₂.

Keywords: nanomicelles, osteosarcoma, radium dichloride, targeted alpha therapy

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Authors: Jae Hong Kim, Sang Cheol Um, Jong Kook Lee

Abstract:

Strong and biocompatible wollastonite (CaSiO3) was fabricated by pressureless sintering at temperature range of 1250~ 1300 ℃ and phase transition of to β-wollastonite with an addition of MgSiO3. Starting pure α-wollastonite powder were prepared by solid state reaction, and MgSiO3 powder was added to α-wollastonite powder to induce the phase transition α to β-wollastonite over 1250℃. Sintered wollastonite samples at 1250℃ with 5 and 10 wt% MgSiO3 were α+β phase and β phase respectively, and showed higher densification rate than that of α or β-wollastonite, which are almost the same as the theoretical density. Hardness and Young’s modulus of sintered wollastonite were dependent on the apparent density and the amount of β-wollastonite. Young’s modulus (78GPa) of β-wollastonite added 10 wt% MgSiO3 was almost double time of sintered α-wollastonite. From the in-vitro test, biphasic (α+β) wollastonite with 5wt% MgSiO3 addition had good bioactivity in simulated body fluid solution.

Keywords: β-wollastonite, high density, MgSiO3, phase transition

Procedia PDF Downloads 565

Authors: Thaís T. Valério Caetano, Lívia de Carvalho Ferreira, João Máximo De Siqueira, Carlos Alexandre Carollo, Arthur Ladeira Macedo, Vanessa C. Stein

Abstract:

Nicotiana glauca, commonly known as wild tobacco or tobacco bush, belongs to the Solanaceae family. It is native to South America but has become naturalized in various regions, including Australia, California, Africa, and the Mediterranean. N. glauca is listed in the Global Invasive Species Database (GISD) and the Invasive Species Compendium (CABI). It is known for producing pyridine alkaloids, including anabasine, which is highly toxic. Anabasine is predominantly found in the leaves and can cause severe health issues such as neuromuscular blockade, respiratory arrest, and cardiovascular problems when ingested. Mistaken identity with edible plants like spinach has resulted in food poisoning cases in Israel and Brazil. Anabasine, a minor alkaloid constituent of tobacco, may contribute to tobacco addiction by mimicking or enhancing the effects of nicotine. Therefore, it is essential to investigate the production pattern of anabasine and its relationship to the developmental stages of the plant. This study aimed to establish the relationship between the phenological plant age, cultivation place, and the increase in anabasine concentration, which can lead to human intoxication cases. In this study, N. glauca plants were collected from three different rural areas in Brazil during a year to examine leaves at various stages of development. Samples were also obtained from cultivated plants in Marilândia, Minas Gerais, Brazil, as well as from Divinópolis, Minas Gerais, Brazil, and Arraial do Cabo, Rio de Janeiro, Brazil. In vitro cultivated plants on MS medium were included in the study. The collected leaves were dried, powdered, and stored. Alkaloid extraction was performed using a methanol and water mixture, followed by liquid-liquid extraction with chloroform. The anabasine content was determined using HPLC-DAD analysis with nicotine as a standard. The results indicated that anabasine production increases with the plant's development, peaking in adult leaves during the reproduction phase and declining afterward. In vitro, plants showed similar anabasine production to young leaves. The successful adaptation of N. glauca in new environments poses a global problem, and the correlation between anabasine production and the plant's developmental stages has been understudied. The presence of substances produced by the plant can pose a risk to other species, especially when mistaken for edible plants. The findings from this study shed light on the pattern of anabasine production and its association with plant development, contributing to a better understanding of the potential risks associated with N. glauca and the importance of accurate identification.

Keywords: alkaloid production, invasive species, nicotiana glauca, plant phenology

Procedia PDF Downloads 60

Authors: K. Khanafer

Abstract:

The aim of this investigation was to validate the fluid-structure interaction (FSI) model of type B aortic dissection with our experimental results from a bench-top-model. Another objective was to study the relationship between the size of a septectomy that increases the outflow of the false lumen and its effect on the values of the differential of pressure between true lumen and false lumen. FSI analysis based on Galerkin’s formulation was used in this investigation to study flow pattern and hemodynamics within a flexible type B aortic dissection model using boundary conditions from our experimental data. The numerical results of our model were verified against the experimental data for various tear size and location. Thus, CFD tools have a potential role in evaluating different scenarios and aortic dissection configurations.

Keywords: aortic dissection, fluid-structure interaction, in vitro model, numerical

Procedia PDF Downloads 257

Authors: M. R. Mogarekar, Shraddha V. More, Pankaj Kumar

Abstract:

Gastroenteritis, the third leading killer of children in India today is responsible for 13% of all deaths in children <5 years of age and kills an estimated 300,000 children in India each year. We decided to investigate parameters which can help in early disease detection and prompt treatment. Serum paraoxonase is calcium dependent esterase which is widely distributed among tissues such as liver, kidney, and intestine and is located in the chromosomal region 7q21.3 22.1. Studies show the presence of excessive reactive oxygen metabolites and antioxidant imbalance in the gastrointestinal tract leading to oxidative stress in gastroenteritis. To our knowledge, this is the first ever study done. The objective of present study is to investigate the role of paraoxonase 1 (PON 1) status i.e arylesterase and lactonase activities and Q192R polymorphism and conjugated dienes, in gastroenteritis of paediatric population. The study and control group consists of 40 paediatric patients with and without gastroenteritis. Paraoxonase arylesterase and lactonase activities were assessed and phenotyping was determined. Conjugated dienes were also assessed. PON 1 arylesterase activities in cases (61.494±13.220) and controls (70.942±15.385) and lactonase activities in cases (15.702±1.036) and controls (17.434±1.176) were significantly decreased (p<0.05). There is no significant difference of phenotypic distribution in cases and controls. Conjugated dienes were found significantly increased in patients (0.086±0.024) than the control group (0.064±0.019) (p<0.05). Paraoxonase 1 activities (arylesterase and lactonase) and conjugated dienes may be useful in risk assessment and management in gastroenteritis in paediatric population.

Keywords: paraoxonase 1 polymorphism, arylesterase, lactonase, conjugated dienes, p-nitrophenylacetate, DHC

Procedia PDF Downloads 293

Authors: Pouya Khaleghi

Abstract:

Dental implants are a successful treatment modality for restoring both function and aesthetics. Dental implant treatment has predictive results in the replacement of the lost teeth and has a high success rate even in the long term. The most important factor which is responsible for the positive course of implant treatment is the process of osseointegration between the implant structure and the host’s bone tissue. During recent years, many studies have focused on surgical and prosthetic factors, as well as the implant-related factors. However, implant failure still occurs despite the improvements that have led to the increased survival rate of dental implants, which suggests the possible role of some host-related risk factors. Vitamin D is a fat-soluble vitamin regulating calcium and phosphorus metabolism in tissues. The role of vitamin D in bone healing has been under investigation for several years. Vitamin D deficiency has also been associated with impaired and delayed callus formation and fractures healing; however, the role of vitamin D has not been clarified. Therefore, it is extremely important to study the phenomenon of a connection formed between bone tissue and the surface of a titanium implant and find correlations between the 25- hydroxycholecalciferol concentration in blood serum and the course of osseointegration. Because the processes of bone remodeling are very dynamic in the period of actual osseointegration, it is necessary to obtain the correct concentration of vitamin D3 metabolites in blood serum. In conclusion, the correct level of 25-hydroxycholecalciferol on the day of surgery and vitamin D deficiency treatment have a significant influence on the increase in the bone level at the implant site during the process of osseointegration assessed radiologically.

Keywords: implant, osseointegration, vitamin d, dental

Procedia PDF Downloads 151

Authors: Sudhir Kumar Sharma, Abiy D. Woldetsadik, Mazin Magzoub, Ramesh Jagannathan

Abstract:

It is well known that bioactive ceramics exhibit specific biological affinities, especially in the area of tissue re-generation. In this context, we report the development of an eminently scalable, novel, supercritical CO₂ based process for the fabrication of hierarchically porous 'soft' CaCO₃ scaffolds. Porosity at the macro, micro, and nanoscales was obtained through process optimization of the so-called 'coffee-ring effect'. Exposure of these CaCO₃ scaffolds to monocytic THP-1 cells yielded negligible levels of tumor necrosis factor-alpha (TNF-α) thereby confirming the lack of immunogenicity of the scaffolds. ECM protein treatment of the scaffolds showed enhanced adsorption comparable to standard control such as glass. In vitro studies using osteoblast precursor cell line, MC3T3, also demonstrated the cytocompatibility of hierarchical porous CaCO₃ scaffolds.

Keywords: supercritical CO2, CaCO3 scaffolds, coffee-ring effect, ECM proteins

Procedia PDF Downloads 282

Authors: Ipsita Biswas, Arnab Sarkar, Ashikur Rahaman, Gopeswar Mukherjee, Subhrangsu Chatterjee, Shamee Bhattacharjee, Deba Prasad Mandal

Abstract:

One of the major reasons for the high mortality rate of lung cancer is the substantial delays in disease detection at late metastatic stages. It is of utmost importance to understand the detailed molecular signaling and detect the molecular markers that can be used for the early diagnosis of cancer. Several studies explored the emerging roles of long noncoding RNAs (lncRNAs) in various cancers as well as lung cancer. A long non-coding RNA LINC00273 was recently discovered to promote cancer cell migration and invasion, and its positive correlation with the pathological stages of metastasis may prove it to be a potential target for inhibiting cancer cell metastasis. Comparing real-time expression of LINC00273 in various human clinical cancer tissue samples with normal tissue samples revealed significantly higher expression in cancer tissues. This long intergenic noncoding RNA was found to be highly expressed in human liver tumor-initiating cells, human gastric adenocarcinoma AGS cell line, as well as human non-small cell lung cancer A549 cell line. SiRNA and shRNA-induced knockdown of LINC00273 in both in vitro and in vivo nude mice significantly subsided AGS and A549 cancer cell migration and invasion. LINC00273 knockdown also reduced TGF-β induced SNAIL, SLUG, VIMENTIN, ZEB1 expression, and metastasis in A549 cells. Plenty of reports have suggested the role of microRNAs of the miR200 family in reversing epithelial to mesenchymal transition (EMT) by inhibiting ZEB transcription factors. In this study, hsa-miR-200a-3p was predicted via IntaRNA-Freiburg RNA tools to be a potential target of LINC00273 with a negative free binding energy of −8.793 kcal/mol, and this interaction was verified as a confirmed target of LINC00273 by RNA pulldown, real-time PCR and luciferase assay. Mechanistically, LINC00273 accelerated TGF-β induced EMT by sponging hsa-miR-200a-3p which in turn liberated ZEB1 and promoted prometastatic functions in A549 cells in vitro as verified by real-time PCR and western blotting. The similar expression patterns of these EMT regulatory pathway molecules, viz. LINC00273, hsa-miR-200a-3p, ZEB1 and TGF-β, were also detected in various clinical samples like breast cancer tissues, oral cancer tissues, lung cancer tissues, etc. Overall, this LINC00273 mediated EMT regulatory signaling can serve as a potential therapeutic target for the prevention of lung cancer metastasis.

Keywords: epithelial to mesenchymal transition, long noncoding RNA, microRNA, non-small-cell lung carcinoma

Procedia PDF Downloads 135

Authors: B. S. Muddukrishna, Karthik Aithal, Aravind Pai

Abstract:

Introduction: Preparation of binary cocrystals of Etraverine (ETR) by using Tartaric Acid (TAR) as a conformer was the main focus of this study. Etravirine is a Class IV drug, as per the BCS classification system. Methods: Cocrystals were prepared by slow evaporation technique. A mixture of total 500mg of ETR: TAR was weighed in molar ratios of 1:1 (371.72mg of ETR and 128.27mg of TAR). Saturated solution of Etravirine was prepared in Acetone: Methanol (50:50) mixture in which tartaric acid is dissolved by sonication and then this solution was stirred using a magnetic stirrer until the solvent got evaporated. Shimadzu FTIR – 8300 system was used to acquire the FTIR spectra of the cocrystals prepared. Shimadzu thermal analyzer was used to achieve DSC measurements. X-ray diffractometer was used to obtain the X-ray powder diffraction pattern. Shake flask method was used to determine the equilibrium dynamic solubility of pure, physical mixture and cocrystals of ETR. USP buffer (pH 6.8) containing 1% of Tween 80 was used as the medium. The pure, physical mixture and the optimized cocrystal of ETR were accurately weighed sufficient to maintain the sink condition and were filled in hard gelatine capsules (size 4). Electrolab-Tablet Dissolution tester using basket apparatus at a rotational speed of 50 rpm and USP phosphate buffer (900 mL, pH = 6.8, 37 ˚C) + 1% Tween80 as a media, was used to carry out dissolution. Shimadzu LC-10 series chromatographic system was used to perform the analysis with PDA detector. An Hypersil BDS C18 (150mm ×4.6 mm ×5 µm) column was used for separation with mobile phase comprising of a mixture of ace¬tonitrile and phosphate buffer 20mM, pH 3.2 in the ratio 60:40 v/v. The flow rate was 1.0mL/min and column temperature was set to 30°C. The detection was carried out at 304 nm for ETR. Results and discussions: The cocrystals were subjected to various solid state characterization and the results confirmed the formation of cocrystals. The C=O stretching vibration (1741cm-1) in tartaric acid was disappeared in the cocrystal and the peak broadening of primary amine indicates hydrogen bond formation. The difference in the melting point of cocrystals when compared to pure Etravirine (265 °C) indicates interaction between the drug and the coformer which proves that first ordered transformation i.e. melting endotherm has disappeared. The difference in 2θ values of pure drug and cocrystals indicates the interaction between the drug and the coformer. Dynamic solubility and dissolution studies were also conducted by shake flask method and USP apparatus one respectively and 3.6 fold increase in the dynamic solubility were observed and in-vitro dissolution study shows four fold increase in the solubility for the ETR: TAR (1:1) cocrystals. The ETR: TAR (1:1) cocrystals shows improved solubility and dissolution as compared to the pure drug which was clearly showed by solid state characterization and dissolution studies.

Keywords: dynamic solubility, Etraverine, in vitro dissolution, slurry method

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Authors: A. Y. Benabdellah, W. Lakhdari, A. Dahliz, Y. Bouchikh, A. Soud, R. M'lik, H. Hammi

Abstract:

The direct comparison tests on the culture medium, between Trichoderma sp. and Fusarium oxysporum f. sp. radicis-lycopersici revealed that the latest one could inhibit the growth of F. oxysporum mycelial over than 40% compared to the control and that after four days of incubation at 26° C. Moreover, beyond this period and at the end of six days, Trichoderma sp. invading the colonies of F. oxysporum on what it sporule, thus revealing its power is highly myco-parasitic. Almost similar results were obtained against Alternaria solani is also a pathogen which is not causing a lot of damage, but we found it more sensitive to Trichoderma sp. with a percentage of inhibition more than 50%. So due to the in vitro test of Trichoderma sp. against these aggressive pathogens by direct contact has been found that can inhibit their mycelial growth with high speed and a high inhibition rate.

Keywords: Trichoderma sp., Fusarium oxysporum f. sp. radicis-lycopersici, Alternaria solani, biological control, antagonist

Procedia PDF Downloads 365

Authors: Tiffany Baetens, Sophie Halliez, Luc Buée, Emiliano Pallecchi, Vincent Thomy, Steve Arscott

Abstract:

We demonstrate here a viable stretchable bio-microelectromechanical systems (BioMEMS) technology for use with biological studies concerned with the effect of high mechanical strains on living cells. An example of this is traumatic brain injury (TBI) where neurons are damaged with physical force to the brain during, e.g., accidents and sports. Robust, miniaturized integrated systems are needed by biologists to be able to study the effect of TBI on neuron cells in vitro. The major challenges in this area are (i) to develop micro, and nanofabrication processes which are based on stretchable substrates and to (ii) create systems which are robust and performant at very high mechanical strain values—sometimes as high as 100%. At the time of writing, such processes and systems were rapidly evolving subject of research and development. The BioMEMS which we present here is composed of an elastomer substrate (low Young’s modulus ~1 MPa) onto which is patterned robust electrodes and insulators. The patterning of the thin films is achieved using standard photolithography techniques directly on the elastomer substrate—thus making the process generic and applicable to many materials’ in based systems. The chosen elastomer used is commercial ‘Sylgard 184’ polydimethylsiloxane (PDMS). It is spin-coated onto a silicon wafer. Multistep ultra-violet based photolithography involving commercial photoresists are then used to pattern robust thin film metallic electrodes (chromium/gold) and insulating layers (parylene) on the top of the PDMS substrate. The thin film metals are deposited using thermal evaporation and shaped using lift-off techniques The BioMEMS has been characterized mechanically using an in-house strain-applicator tool. The system is composed of 12 electrodes with one reference electrode transversally-orientated to the uniaxial longitudinal straining of the system. The electrical resistance of the electrodes is observed to remain very stable with applied strain—with a resistivity approaching that of evaporated gold—up to an interline strain of ~50%. The mechanical characterization revealed some interesting original properties of such stretchable BioMEMS. For example, a Poisson effect induced electrical ‘self-healing’ of cracking was identified. Biocompatibility of the commercial photoresist has been studied and is conclusive. We will present the results of the BioMEMS, which has also characterized living cells with a commercial Multi Electrode Array (MEA) characterization tool (Multi Channel Systems, USA). The BioMEMS enables the cells to be strained up to 50% and then characterized electrically and optically.

Keywords: BioMEMS, elastomer, electrical impedance measurements of living cells, high mechanical strain, microfabrication, stretchable systems, thin films, traumatic brain injury

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Authors: Lucian Mocan, Teodora Mocan, Matea Cristian, Cornel Iancu

Abstract:

We present method of nanoparticle enhanced laser thermal ablation of Klebsiella pneumoniae infections, using gold nanoparticles combined with a specific growth factor and demonstrate its selective therapeutic efficacy. Ab (antibody solution) bound to GNPs (gold nanoparticles) was administered in vitro and determined the specific delivery of the nano-bioconjugate into the microorganism. The extent of necrosis was considerable following laser therapy, and at the same time, normal cells were not seriously affected. The selective photothermal ablation of the infected tissue was obtained after the selective accumulation of Ab bound to GNPs into bacteria following perfusion. These results may represent a major step in antibiotherapy treatment using nanolocalized thermal ablation by laser heating.

Keywords: gold nanoparticles, Klebsiella pneumoniae, nanoparticle functionalization, laser irradiation, antibody

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Authors: Marek Gancarz, Agnieszka Nawrocka, Robert Rusinek, Marcin Tadla

Abstract:

Volatile organic compounds (VOCs) are the fungi metabolites in the gaseous form produced during improper storage of agricultural commodities (e.g. grain, food). The spoilt commodities produce a wide range of VOCs including alcohols, esters, aldehydes, ketones, alkanes, alkenes, furans, phenols etc. The characteristic VOCs and odours can be determined by using electronic nose (e-Nose) which contains a matrix of different kinds of sensors e.g. resistive sensors. The aim of the present studies was to determine relationship between response of the resistive sensors on the chosen volatiles and their concentration. According to the literature, it was chosen volatiles characteristic for the cereals: ethanol, 3-methyl-1-butanol and hexanal. Analysis of the sensor signals shows that a signal shape is different for the different substances. Moreover, each VOC signal gives information about a maximum of the normalized sensor response (R/Rmax), an impregnation time (tIM) and a cleaning time at half maximum of R/Rmax (tCL). These three parameters can be regarded as a ‘VOC fingerprint’. Seven resistive sensors (TGS2600-B00, TGS2602-B00, TGS2610-C00, TGS2611-C00, TGS2611-E00, TGS2612-D00, TGS2620-C00) produced by Figaro USA Inc., and one (AS-MLV-P2) produced by AMS AG, Austria were used. Two out of seven sensors (TGS2611-E00, TGS2612-D00) did not react to the chosen VOCs. The most responsive sensor was AS-MLV-P2. The research was supported by the National Centre for Research and Development (NCBR), Grant No. PBS2/A8/22/2013.

Keywords: agricultural commodities, organic compounds, resistive sensors, volatile

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Authors: Shivankar Agrawal, Indira Sarangthem

Abstract:

Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus (MRSA) remains one of the major causes of healthcare-associated and community-onset infections worldwide. Hence the search for non-toxic natural compounds having antibacterial activity has intensified for future drug development. The exploration of less studied niches of Earth can highly increase the possibility to discover novel bioactive compounds. Therefore, in this study, the cultivable fraction of fungi from the sediments of natural hot springs has been studied to mine potential fungal candidates with antibacterial activity against the human pathogen Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. We isolated diverse strains of thermophilic fungi from a collection of samples from sediment. Following a standard method, we isolated a promising thermophilic fungus strain IBSD19, identified as Acrophialophora levis, possessing the potential to produce an anti-Staphylococcus aureus agent. The growth conditions were optimized and scaled to fermentation, and its produced extract was subjected to chemical extraction. The ethyl acetate fraction was found to display significant activity against Staphylococcus aureus and MRSA with a minimum inhibitory concentration (MIC) of 0.5 mg/ml and 4 mg/ml, respectively. The cell membrane integrity assay and SEM suggested that the fungal metabolites cause bacteria clustering and further lysis of the cell.

Keywords: antibacterial activity, antioxidant, fungi, Staphylococcus aureus, MRSA, thermophiles

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Authors: L. Mallesha, C. S. Karthik, P. Mallu

Abstract:

A series of new [1-(substituted-benzoyl)-piperidin-4-yl]-(2,4-difluoro-phenyl)-methanone oxime derivatives, 3(a-f) were synthesized and characterized by different spectral studies. All compounds were evaluated for their in vitro antibacterial activity against bacterial strains. These compounds were screened for their antioxidant activity by DPPH• and Fe2+ chelating assay. Antiproliferative effects were evaluated using the MTT assay method against two human cancer cell lines and one astrocytoma brain tumor cell line. Compound 3b exhibited moderate antibacterial activity when compared with other compounds. All the compounds showed antioxidant activity, where compound 3f was the best radical scavenger and Fe2+ ion scavenger. Compounds, 3b, and 3d showed good activity on all cell lines, whereas the other compounds in the series exhibited moderate activity.

Keywords: Piperidine, antibacterial, antioxidant, antiproliferative

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Authors: Amruta D. S. Pathare, Indira Hinduja, Kusum Zaveri

Abstract:

Implantation failure (IF) even after the good-quality embryo transfer (ET) in the physiologically normal endometrium is the main obstacle in in vitro fertilization (IVF). Various microarray studies have been performed worldwide to elucidate the genes requisite for endometrial receptivity. These studies have included the population based on different phases of menstrual cycle during natural cycle and stimulated cycle in normal fertile women. Additionally, the literature is also available in recurrent implantation failure patients versus oocyte donors in natural cycle. However, for the first time, we aim to study the genomics of endometrial receptivity in IF patients under controlled ovarian stimulation (COS) during which ET is generally practised in IVF. Endometrial gene expression profiling in IF patients (n=10) and oocyte donors (n=8) were compared during window of implantation under COS by whole genome microarray (using Illumina platform). Enrichment analysis of microarray data was performed to determine dys-regulated biological functions and pathways using Database for Annotation, Visualization and Integrated Discovery, v6.8 (DAVID). The enrichment mapping was performed with the help of Cytoscape software. Microarray results were validated by real-time PCR. Localization of genes related to immune response (Progestagen-Associated Endometrial Protein (PAEP), Leukaemia Inhibitory Factor (LIF), Interleukin-6 Signal Transducer (IL6ST) was detected by immunohistochemistry. The study revealed 418 genes downregulated and 519 genes upregulated in IF patients compared to healthy fertile controls. The gene ontology, pathway analysis and enrichment mapping revealed significant downregulation in activation and regulation of immune and inflammation response in IF patients under COS. The lower expression of Progestagen Associated Endometrial Protein (PAEP), Leukemia Inhibitory Factor (LIF) and Interleukin 6 Signal Transducer (IL6ST) in cases compared to controls by real time and immunohistochemistry suggests the functional importance of these genes. The study was proved useful to uncover the probable reason of implantation failure being imbalance of immune and inflammatory regulation in our group of subjects. Based on the present study findings, a panel of significant dysregulated genes related to immune and inflammatory pathways needs to be further substantiated in larger cohort in natural as well as stimulated cycle. Upon which these genes could be screened in IF patients during window of implantation (WOI) before going for embryo transfer or any other immunological treatment. This would help to estimate the regulation of specific immune response during WOI in a patient. The appropriate treatment of either activation of immune response or suppression of immune response can be then attempted in IF patients to enhance the receptivity of endometrium.

Keywords: endometrial receptivity, immune and inflammatory response, gene expression microarray, window of implantation

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Authors: Ramesh Subramani, Mathivanan Narayanasamy, William Aalbersberg

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It is well accepted by last 35 years of research and on-going programmes that marine environment harbours abundant and unique biodiversity, which is currently playing as an important source in bioprospecting. It has become apparent that marine microorganisms are lead in the biodiscovery. Among marine organisms, actinobacteria are a target phylum for discovering novel antibiotics against increasing the multi-drug resistant human pathogens because of these taxa representing for novel genera and species. Marine actinomycetes are a proven source of new antibiotic leads and novel enzymes with important industrial applications. A total of 183 streptomycete and 25 non-streptomycete strains were isolated from different marine samples collected from north-eastern part of the Indian Ocean. Among them, 111 isolates displayed antibacterial activity against human pathogens and 151 exhibited antifungal activity against phytopathogens. Importantly, most of them produced various extracellular enzymes and 58 of them produced exopolysaccharides. Totally eight small bioactive compounds and a thermostable alkaline protease have been purified from a selected strain, Streptomyces fungicidicus. Besides, our on-going studies on non-streptomycete strains (rare actinomycetes) are most likely promising resource for new and unique compounds against current emerging drug-resistant pathogens. We have just recognised the chemical diversity in marine microorganisms. Therefore it is worthwhile to continue the exploration of marine microorganisms for new drug leads, novel enzymes and other bioprospecting research.

Keywords: bioactive compounds, industrial enzymes, marine actinobacteria, microbial metabolites, marine natural products

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Authors: Gurpreet Singh, Hazoor Singh, Buta Singh Sidhu

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In the current investigation plasma spray technique was used for depositing hydroxyapatite (HA) and HA – silicon oxide (SiO2) coatings on 316L SS substrate. In HA-SiO2 coating, 20 wt% SiO2 was mixed with HA. The feedstock and coatings were characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM)/energy-dispersive X-ray spectroscopy (EDX) analyses. The corrosion resistance of the uncoated, HA coated and HA + 20 wt% SiO2 coated 316L SS was investigated by electrochemical corrosion testing in simulated human body fluid (Ringer’s solution). The influence of SiO2 (20 wt%) on corrosion resistance was determined. After the corrosion testing, the samples were analyzed by XRD and SEM/EDX analyses. The addition of SiO2 reduces the crystallinity of the coating. The corrosion resistance of the 316L SS was found to increase after the deposition of the HA + 20 wt% SiO2 and HA coatings.

Keywords: HA, SiO2, corrosion, Ringer’s solution, 316L SS

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Authors: J. Herrero, D. Puigserver, I. Nijenhuis, K. Kuntze, J. M. Carmona

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In the transition zone between aquifers and basal aquitards, the perchloroethene (PCE)-pools are more recalcitrant than those elsewhere in the aquifer. Although biodegradation of chloroethenes occur in this zone, it is a slow process and a remediation strategy is needed. The aim of this study is to demonstrate that combined strategy of biostimulation and in situ chemical reduction (ISCR) is more efficient than the two separated strategies. Four different microcosm experiments with sediment and groundwater of a selected field site where an aged pool exists at the bottom of a transition zone were designed under i) natural conditions, ii) biostimulation with lactic acid, iii) ISCR with zero-value iron (ZVI) and under iv) a combined strategy with lactic acid and ZVI. Biotic and abiotic dehalogenation, terminal electron acceptor processes and evolution of microbial communities were determined for each experiment. The main results were: i) reductive dehalogenation of PCE-pools occurs under sulfate-reducing conditions; ii) biostimulation with lactic acid supports more pronounced reductive dehalogenation of PCE and trichloroethene (TCE), but results in an accumulation of 1,2-cis-dichloroethene (cDCE); iii) ISCR with ZVI produces a sustained dehalogenation of PCE and its metabolites iv) combined strategy of biostimulation and ISCR results in a fast dehalogenation of PCE and TCE and a sustained dehalogenation of cisDCE. These findings suggest that biostimulation and ISCR with ZVI are the most suitable strategies for a complete reductive dehalogenation of PCE-pools in the transition zone and further to enable the dissolution of dense non-aqueous phase liquids.

Keywords: aged PCE-pool, anaerobic microcosm experiment, biostimulation, in situ chemical reduction, natural attenuation

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Authors: Ali Mirtaghavi, Andy Baldwin, Rajendarn Muthuraj, Jack Luo

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Recent advances in biomaterials have led to utilizing biopolymers to develop 3D scaffolds in tissue regeneration. One of the major challenges of designing biomaterials for 3D scaffolds is to mimic the building blocks similar to the extracellular matrix (ECM) of the native tissues. Biopolymer based aerogels obtained by freeze-drying have shown to provide structural similarities to the ECM owing to their 3D format and a highly porous structure with interconnected pores, similar to the ECM. Gelatin (GEL) is known to be a promising biomaterial with inherent regenerative characteristics owing to its chemical similarities to the ECM in native tissue, biocompatibility abundance, cost-effectiveness and accessible functional groups, which makes it facile for chemical modifications with other biomaterials to form biocomposites. Despite such advantages, gelatin offers poor mechanical properties, sensitive enzymatic degradation and high viscosity at room temperature which limits its application and encourages its use to develop biocomposites. Hydrophilic biomass-based cellulose nanofibrous (CNF) has been explored to use as suspension for biocomposite aerogels for the development of 3D porous structures with excellent mechanical properties, biocompatibility and slow enzymatic degradation. In this work, CNF biocomposite aerogels with various ratios of CNF:GEL) (90:10, 70:30 and 50:50) were prepared by freeze-drying technique, and their properties were investigated in terms of physicochemical, mechanical and biological characteristics. Epichlorohydrin (EPH) was used to investigate the effect of chemical crosslinking on the molecular interaction of CNF: GEL, and its effects on physicochemical, mechanical and biological properties of the biocomposite aerogels. Ultimately, chemical crosslinking helped to improve the mechanical resilience of the resulting aerogels. Amongst all the CNF-GEL composites, the crosslinked CNF: GEL (70:30) biocomposite was found to be favourable for cell attachment and viability. It possessed highly porous structure (porosity of ~93%) with pore sizes ranging from 16-110 µm, adequate mechanical properties (compression modulus of ~47 kPa) and optimal biocompatibility both in-vitro and in-vivo, as well as controlled enzymatic biodegradation, high water penetration, which could be considered a suitable option for wound healing application. In-vivo experiments showed improvement on inflammation and foreign giant body cell reaction for the crosslinked CNF: GEL (70:30) compared to the other samples. This could be due to the superior interaction of CNF with gelatin through chemical crosslinking, resulting in more optimal in-vivo improvement. In-vitro cell culture investigation on human dermal fibroblasts showed satisfactory 3D cell attachment over time. Overall, it has been observed that the developed CNF: GEL aerogel can be considered as a potential scaffold for soft tissue regeneration application.

Keywords: 3D scaffolds, aerogels, Biocomposites , tissue engineering

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Authors: Asghar Ebrahimzadeh, Sattar Malekian, Mohammad Ali Aazami, Mohammad Bagher Hassanpouraghdam

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Roses are of main ornamental flowers worldwide. Rosa damascena Mill., besides being an ornamental plant, has major pharmaceutical, cosmetic and fragrance applications. Traditional propagation methods of the plant are using suckers, cutting and grafting. In the present experiment, we used the different explants (leaf section, petioles and nodal cutting) for the optimization of this high-valued ornamental from a native clonal plant. Diverse explants were acquired from mature plants during the growing season and were planted on MS medium supplemented with different hormonal combinations. 70% alcohol and sodium hypochloride were utilized for the surface sterilization. For proliferation, BAP and BA (1-5 mg L-1) and NAA (1-2 mg L-1) were tested. The highest proliferation rate was afforded from MS medium supplemented with 1.5 mg L-1 BA and 5 mg L-1 BAP. Callogenesis from leaf samples and petioles was the best with 1/2 MS medium enriched with 1mg L-1 BAP and 4 mg L-1 2,4-D. Rooting was occurred with the highest frequency in a medium containing 0.1 mg L-1 IBA.

Keywords: Rosa damascene, micropropagation, petiole, IBA, BAP

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Authors: Yuva Bellik, Selles Mohamed Amar

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The beneficial health effects including antioxidant properties of mint (Mentha spicata) and honey bees (Apis mellifera) have been extensively studied. However, there is no data about the effects of their associated use. In this study the total phenolic and flavonoid contents for individual extracts of mint and honey and their combination were determined. The antioxidant activity was investigated by using reducing power, 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2´- azinobis-(3-ethylbenzothiazoline-6-sulphonic acid diamonium salt (ABTS), and chelating power methods. The results showed that individual extracts contained important quantity of phenolics and flavonoids and their combination was found to produce best antioxidant activity. A significant linear correlation between the phenolic/flavonoid contents and antioxidant activity, especially with reducing power and free radical scavenging abilities, was observed.

Keywords: honey, mint, synergy, antioxidant activity

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Authors: Labiba Zerari Bourafa, Djebar Mohamed Reda, Berrebah Houria, Khadri Sihem, Chiheb Linda

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The study of the effect of metal dust (pollutants) was performed on higher plant white beans Phaseolus vulgaris; the experience took place in cellular toxicology laboratory (in vitro culture). The seeds of the bean Phaseolus vulgaris are cultured in a metal contaminated dust medium (a single treatment by different increasing doses), at a rate of 10 seeds per box, for 10 days. The measurement of morpho-metric parameters is performed during the first 96 hours that follow the germination; while the dosage of the proline, the protein content and histological sections are formed on the tenth day (240 h). All morpho-metric and biochemical parameters measured were highly disturbed by metal dust; histological sections confirm this disurbance.

Keywords: conductive fabrics, metal dust, osmoticums, roots, Phaseolus vulgaris

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Authors: Ryan D. Martinus

Abstract:

Inflammation of the endothelium is an important process leading to diabetic atherosclerosis. However, the molecular mechanisms by which diabetes contributes to endothelial inflammation remain to be established. Using In-vitro cultured Human cells and Hsp60 specific ELISA assays, we show that Hsp60 is not only induced in Human monocyte cells under hyperglycaemic conditions but that the Hsp60 is also secreted from these cells. Furthermore, we also demonstrate that the Hsp60 secreted from these monocyte cells is also able to activate Toll-like receptor-4 (TLR4) from Human endothelial cells. This suggests that a potential link may exist between the hyperglycaemia-induced expression of Hsp60 in monocyte cells and vascular inflammation. Circulating levels of Hsp60 due to mitochondrial stress in diabetes patients could, therefore, be an important modulator of inflammation in endothelial cells and thus contribute to the increased incidences of atherosclerosis in diabetes mellitus.

Keywords: mitochondria, Hsp60, inflammation, diabetes mellitus

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Authors: Awatif Boumaza, Abderraouf Hilali, Hayat Talbi, Houda Sbayou

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The methanolic extract of Zygophyllum cornutun coss, an Algerian medicinal plant, was screened to the presence of mutagenic activity and genotoxic effect using the Ames test (Salmonella/microsome) and the micronucleus assay respectively. Positive results were obtained with both tests. The Ames test showed mutagenic activity in the presence of microsomal activation, while negative result was observed without microsomal activation. In the micronucleus test, two parameters were evaluated: the frequency of the micronucleus that increased in a dose dependent way and the proliferation index that decreased according to the micronucleus frequency. Even that further studies must be carried out, the mutagenic activity and the genotoxic effect of Zygophyllum cornutum should be taken in consideration when used as therapeutic plant.

Keywords: ames test, micronucleus test, mutagenic activity, genotoxicity, Zygophyllum cornutum

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Authors: M. Nerantzaki, I. Koliakou, D. Bikiaris

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This study evaluates the hypothesis that the incorporation of fibrous hydroxyapatite nanoparticles (nHA) with high crystallinity and high aspect ratio, synthesized by hydrothermal method, into Poly(butylene succinate) (PBSu), improves the bioactivity of the aliphatic polyester and affects new bone growth inhibiting resorption and enhancing bone formation. Hydroxyapatite nanorods were synthesized using a simple hydrothermal procedure. First, the HPO42- -containing solution was added drop-wise into the Ca2+-containing solution, while the molar ratio of Ca/P was adjusted at 1.67. The HA precursor was then treated hydrothermally at 200°C for 72 h. The resulting powder was characterized using XRD, FT-IR, TEM, and EDXA. Afterwards, PBSu nanocomposites containing 2.5wt% (nHA) were prepared by in situ polymerization technique for the first time and were examined as potential scaffolds for bone engineering applications. For comparison purposes composites containing either 2.5wt% micro-Bioglass (mBG) or 2.5wt% mBG-nHA were prepared and studied, too. The composite scaffolds were characterized using SEM, FTIR, and XRD. Mechanical testing (Instron 3344) and Contact Angle measurements were also carried out. Enzymatic degradation was studied in an aqueous solution containing a mixture of R. Oryzae and P. Cepacia lipases at 37°C and pH=7.2. In vitro biomineralization test was performed by immersing all samples in simulated body fluid (SBF) for 21 days. Biocompatibility was assessed using rat Adipose Stem Cells (rASCs), genetically modified by nucleofection with DNA encoding SB100x transposase and pT2-Venus-neo transposon expression plasmids in order to attain fluorescence images. Cell proliferation and viability of cells on the scaffolds were evaluated using fluoresce microscopy and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) assay. Finally, osteogenic differentiation was assessed by staining rASCs with alizarine red using cetylpyridinium chloride (CPC) method. TEM image of the fibrous HAp nanoparticles, synthesized in the present study clearly showed the fibrous morphology of the synthesized powder. The addition of nHA decreased significantly the contact angle of the samples, indicating that the materials become more hydrophilic and hence they absorb more water and subsequently degrade more rapidly. In vitro biomineralization test confirmed that all samples were bioactive as mineral deposits were detected by X-ray diffractometry after incubation in SBF. Metabolic activity of rASCs on all PBSu composites was high and increased from day 1 of culture to day 14. On day 28 metabolic activity of rASCs cultured on samples enriched with bioceramics was significantly decreased due to possible differentiation of rASCs to osteoblasts. Staining rASCs with alizarin red after 28 days in culture confirmed our initial hypothesis as the presence of calcium was detected, suggesting osteogenic differentiation of rACS on PBSu/nHAp/mBG 2.5% and PBSu/mBG 2.5% composite scaffolds.

Keywords: biomaterials, hydroxyapatite nanorods, poly(butylene succinate), scaffolds

Procedia PDF Downloads 292

Authors: Katarína Koňariková, Georgios A. Perdikaris, Lucia Andrezálová, Zdeňka Ďuračková, Lucia Laubertová, Helena Gbelcová, Ingrid Žitňanová

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Introduction: The continuous demand for new anti-cancer drugs has stimulated chemotherapeutic research based on the use of essential metalloelements with the aim to develop potential drugs with lower toxicity and higher antiproliferative activity against tumors. Copper(II) and its complexes play an important role as suitable species for antiproliferative tests. Objectives: The central objective of the current study was to investigate the potential in vitro anti-proliferative effects of N-salicylidene-L-glutamato copper (II) complexes and molecular mechanism of apoptosis induced by tested complexes. In our project we tested N-salicylidene-L-glutamato copper (II) complexes ZK1 - [Cu(N-salicylidene-L-glutamato)(H2O)2].H2O; MK0 - ([Cu2(N-sal-D,L-glu)2(isoquinoline)2].2H2O); MK1 - [Cu(N-salicylidene-5-methyl-L-glutamato)(H2O)].H2O; MK3 - transbis(ethanol)tetrakis(imidazol)Cu(II)(2+)bis(N-salicylidene-D,L-glutamato-N,O)-KO:KO´-(imidazol); MK5 - [Cu(N-salicylidene-D,L- glutamato)(2-methylimidazol] at concentration range 0.001-100 µmol/L against human colon carcinoma cell line HT-29 and human breast carcinoma cell line MCF-7. Methods: Viability was assessed by direct counting of 0.4% trypan blue dye-excluding cells after 24, 48 and 72 hour cultivations with or without copper complex and by MTT assay. To analyze the type of cell death and its mechanism induced by our copper complex we used different methods. To distinguish apoptosis from necrosis we used electrophoretic analysis, to study the activity of caspases 8 and 9 – luminometric analysis and caspase activity 3 colorimetric assay. Results: The observed anti-proliferative effect of the copper complexes appeared to be dose-, time- and cell line- dependent. Human colon carcinoma cells HT-29 appeared to be more sensitive to the complex MK0 ([Cu2(N-sal-D,L-glu)2(isoquinoline)2].2H2O) than to ZK1 ([Cu(N-salicylidene-L-glutamato)(H2O)2].H2O) and MK1 ([Cu(N-salicylidene-5-methyl-L-glutamato)(H2O)].H2O)). Human colon carcinoma cells HT-29 appeared to be more sensitive to the complex than human breast carcinoma cells MCF-7. IC50 decreased with time of incubation (24, 48 and 72h) for HT-29, but increased for MCF-7. By electrophoresis we found apoptotic cell death induced by our copper complexes in HT-29 at concentrations 1, 10, 50 and 100 µmol/L after 48h (ZK1) and 72h (MK0, MK1) and in MCF-7 we did not find apoptosis. We also studied molecular mechanism of apoptosis in HT-29 induced by copper complexes. We found active caspase 9 in HT-29 after ZK1 ([Cu(N-salicylidene-L-glutamato)(H2O)2].H2O) and MK1 ([Cu(N-salicylidene-5-methyl-L-glutamato)(H2O)].H2O)) influence and active caspase 8 after MK0 ([Cu2(N-sal-D,L-glu)2(isoquinoline)2].2H2O) influence. Conclusion: Our copper complexes showed cytotoxic activities against human colon carcinoma cells HT-29 and breast cancer cell line MCF-7 in vitro. Apoptosis was activated by mitochondrial pathway (intrinsic pathway) in case of ZK1 [Cu(N-salicylidene-L-glutamato)(H2O)2].H2O; MK1 [Cu(N-salicylidene-5-methyl-L-glutamato)(H2O)].H2O; MK3 - transbis(ethanol)tetrakis(imidazol)Cu(II)(2+)bis(N-salicylidene-D,L-glutamato-N,O)-KO:KO´-(imidazol) and MK5 - [Cu(N-salicylidene-D,L- glutamato)(2-methylimidazol] copper complexes and by death receptors (extrinsic pathway) in case of MK0 [Cu2(N-sal-D,L-glu)2(isoquinoline)2].2H2O copper complex in HT-29.

Keywords: apoptosis, copper complex, cancer, carcinoma cell line

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Authors: Anne Elain, Magali Le Fellic

Abstract:

Halogenated Aromatic Compounds (HAC) are major organic pollutants that are detected in several environmental compartments as a result of their widespread use as solvents, pesticides and other industrial chemicals. The degradation of HAC simultaneously at low temperature and under saline conditions would be useful for remediation of polluted sites. Hence, microbial processes based on the metabolic activities of anaerobic bacteria are especially attractive from an economic and environmental point of view. Metabolites are generally less toxic, less likely to bioaccumulate and more susceptible for further degradation. Studies on biological reductive dehalogenation have largely been restricted to chlorinated compounds while relatively few have focussed on other HAC i.e., fluorinated, brominated or iodinated compounds. The objectives of the present work were to investigate the biodegradation of a mixture of triiodoaromatic molecules in industrial wastewater by an enriched bacterial consortium. Biodegradation of the mixture was studied during batch experiments in an anaerobic reactor. The degree of mineralization and recovery of halogen were monitored by HPLC-UV, TOC analysis and potentiometric titration. Providing ethanol as an electron donor was found to stimulate anaerobic reductive dehalogenation of HAC with a deiodination rate up to 12.4 mg.L-1 per day. Sodium chloride even at high concentration (10 mM) was found to have no influence on the degradation rates nor on the microbial viability. An analysis of the 16S rDNA (MicroSeq®) revealed that at least 6 bacteria were predominant in the enrichment, including Pseudomonas aeruginosa, Pseudomonas monteilii, Kocuria rhizophila, Ochrobacterium anthropi, Ralstonia pickettii and Rhizobium rhizogenes.

Keywords: halogenated aromatics, anaerobic biodegradation, deiodination, bacterial consortium

Procedia PDF Downloads 157