Search results for: In vitro cytotoxicity

Authors: Ezzati Givi M., Hoveizi E., Nezhad Marani N.

Abstract:

Platinum-based anticancer therapeutics are the most widely used drugs in clinical chemotherapy but have major limitations and various side effects in clinical applications. Gonadotoxicity and sterility is one of the most common complications for cancer survivors, which seem to be drug-specific and dose-related. Therefore, many efforts have been dedicated to discovering a new structure of platinum-based anticancer agents with improved therapeutic index, fewer side effects. In this regard, new Pt(II)-phosphane complexes containing heterocyclic thionate ligands (PCTL) have been synthesized, which show more potent antitumor activities in comparison to cisplatin. Cisplatin, the best leading metal-based antitumor drug in the field, induces testicular toxicity on Leydig and Sertoli cells leading to serious side effects such as azoospermia and infertility. Therefore in the present study, we aimed to investigate the cytotoxicity effect of PCTL on mice TM4 Sertoli cells with particular emphasis on the role of autophagy in comparison to cisplatin. In this study, an MTT assay was performed to evaluate the IC50 of PCTL and to analyze the TM3 Leydig cell's viability. Cells morphology was evaluated via invert microscope and Changing in morphology for nuclei swelling or autophagic vacuoles formation were assessed by DAPI and MDC staining. Testosterone production in the culture medium was measured using an ELISA kit. Finally, the expression of Autophagy-related genes, Atg5, Beclin1 and p62, were analyzed by qPCR. Based on the obtained results by MTT, the IC50 value of PCTL was 50 μM in TM3 cells and cytotoxic effects was in a dose- and time-dependent manner. Cells morphological changes investigated by inverted microscopy, DAPI, and MDC staining which showed the cytotoxic concentrations of PCTL was significantly higher than cisplatin in the treated TM3 Leydig cells. The results of PCR showed a lack of expression of the p62, Atg5 and Beclin1 gene in TM3 cells treated with PCTL in comparison to cisplatin and control groups. It should be noted that the effects of 25 μM PCTL concentration on TM3 cells have been associated with increased testosterone production and secretion, which requires further study to explain the possible causes and involved molecular mechanisms. The results of the study showed that the PCTL had less-lethal effects on TM3 cells in comparison to cisplatin and probably did not induce autophagy in TM3 cells.

Keywords: platinum-based anticancer agents, cisplatin, Leydig TM3 cells, autophagy

Procedia PDF Downloads 128

Authors: Haidy N. Salem, Nada O. Kamel, Shahinaz N. Hassan, Sherif M. Elhefnawy

Abstract:

Aim: to assess the effect of using turmeric powder extract on changes of tooth color with extra-coronal and intra-coronal bleaching methods. Methods: Turmeric powder extract was weighted and mixed with two different hydrogen peroxide concentrations (3% and 6%) to be used as a bleaching agent. Thirty teeth were allocated into three groups (n=10): Group A: Bleaching agent (6%) was applied on the labial surface, Group B: Bleaching agent (3%) was applied inside the pulp chamber and Group C: Extra and intra-coronal bleaching techniques were used (6% and 3% respectively). A standardized access cavity was opened in the palatal surface of each tooth in both Groups B and C. Color parameters were measured using a spectrophotometer. Results: A statistically significant difference in color difference values (∆E*) and enamel brightness (∆L*) was found between Group C and each of Groups A and B. There was no statistically significant difference in (∆E*) and (∆L*) between Group A and Group B. The highest mean value of (∆E*) and (∆L*) was found in Group C, while the least mean value was found in Group B. Conclusion: Bleaching the external and internal tooth structure with low concentrations of hydrogen peroxide solution mixed with turmeric extract has a promising effect in color enhancement.

Keywords: bleaching, hydrogen peroxide, spectrophotometer, turmeric

Procedia PDF Downloads 118

Authors: Hachul Jung, Ahee Kim, Sanghoon Lee, Dahye Kwon, Songwoo Yoon, Jinhee Moon

Abstract:

We fabricated a wearable patch device including novel patch type flexible dry electrode based on carbon nanofibers (CNFs) and silicone-based elastomer (MED 6215) for real-time ECG monitoring. There are many methods to make flexible conductive polymer by mixing metal or carbon-based nanoparticles. In this study, CNFs are selected for conductive nanoparticles because carbon nanotubes (CNTs) are difficult to disperse uniformly in elastomer compare with CNFs and silver nanowires are relatively high cost and easily oxidized in the air. Wearable patch is composed of 2 parts that dry electrode parts for recording bio signal and sticky patch parts for mounting on the skin. Dry electrode parts were made by vortexer and baking in prepared mold. To optimize electrical performance and diffusion degree of uniformity, we developed unique mixing and baking process. Secondly, sticky patch parts were made by patterning and detaching from smooth surface substrate after spin-coating soft skin adhesive. In this process, attachable and detachable strengths of sticky patch are measured and optimized for them, using a monitoring system. Assembled patch is flexible, stretchable, easily skin mountable and connectable directly with the system. To evaluate the performance of electrical characteristics and ECG (Electrocardiography) recording, wearable patch was tested by changing concentrations of CNFs and thickness of the dry electrode. In these results, the CNF concentration and thickness of dry electrodes were important variables to obtain high-quality ECG signals without incidental distractions. Cytotoxicity test is conducted to prove biocompatibility, and long-term wearing test showed no skin reactions such as itching or erythema. To minimize noises from motion artifacts and line noise, we make the customized wireless, light-weight data acquisition system. Measured ECG Signals from this system are stable and successfully monitored simultaneously. To sum up, we could fully utilize fabricated wearable patch devices for real-time ECG monitoring easily.

Keywords: carbon nanofibers, ECG monitoring, flexible dry electrode, wearable patch

Procedia PDF Downloads 185

Authors: Niranjan Koirala, Sumangala Darsandhari, Hye Jin Jung, Jae Kyung Sohng

Abstract:

Sakuranetin, an antifungal agent and genkwanin, an anti-inflammatory agent, are flavonoids with several potential pharmaceutical applications. To produce such valuable flavonoids in large quantity, an Escherichia coli cell factory has been created. E. coli harboring O-methyltransferase (SaOMT2) derived from Streptomyces avermitilis was employed for regiospecific methylation of naringenin and apigenin. In order to increase the production via biotransformation, metK gene was overexpressed and the conditions were optimized. The maximum yield of sakuranetin and genkwanin under optimized conditions was 197 µM and 170 µM respectively when 200 µM of naringenin and apigenin were supplemented in the separate cultures. Furthermore, sakuranetin was purified in large scale and used as a substrate for in vitro glycosylation by YjiC to produce glucose and galactose derivatives of sakuranetin for improved solubility. We also found that unlike naringenin, sakuranetin effectively inhibits α-melanocyte stimulating hormone (α-MSH)-stimulated melanogenesis in B16F10 melanoma cells. In addition, genkwanin more potently inhibited angiogenesis than apigenin. Based on our findings, we speculate that these compounds warrant further investigation in vivo as potential new therapeutic anti-carcinogenic, anti-melanogenic and anti-angiogenic agents.

Keywords: anti-carcinogenic, anti-melanogenic, glycosylation, methylation

Procedia PDF Downloads 609

Authors: Oluwafemi Ojo, Omotade Oloyede

Abstract:

This study seeks to investigate the antioxidative properties and the ability of aqueous, ethanolic and ethyl acetate extracts from Ocimum gratissimum (OG) leaves to inhibit some pro-oxidants (Fe2+ and sodium nitroprusside) induced lipid peroxidation in rat’s liver homogenates in vitro. The ability of the extracts to inhibit 25 µM FeSO4 and 7.0 µM sodium nitroprusside induced lipid peroxidation in isolated rat’s liver was determined. The results of the study revealed that both pro-oxidants caused a significantly decrease in (p < 0.05) accumulation of lipid peroxides. However, aqueous extract of OG shows a high ability to inhibit lipid production in the liver induced with SNP than Fe2+. Ethanolic and ethyl acetate extract of OG which shows a high ability to inhibit lipid production more when induced with Fe2+ than SNP. However, ethyl acetate fraction of OG shows a higher inhibitory effect on both Fe2+ and SNP induced lipid peroxidation in rat’s liver. This applies to its significantly higher extractable phytochemicals. Therefore, Fe II and sodium nitroprusside induced oxidative stress could be managed by dietary intake of Ocimum gratissimum leaves.

Keywords: antioxidative, pro-oxidants, lipid peroxidation, Ocimum gratissimum

Procedia PDF Downloads 479

Authors: Ashish Jain, Satish Nayak

Abstract:

Transdermal iontophoretic drug delivery system is viable drug delivery platform technology and has a strong market worldwide. Transdermal drug delivery system is particularly desirable for therapeutic agents that need prolonged administration at controlled plasma level. This makes appropriateness to antihypertensive and anti-diabetic agents for their transdermal development. Controlled zero order absorption, easily termination of drug delivery and easy to administration also support for popularity of transdermal delivery. In this current research iontophoretic delivery of various anti diabetic agents like glipizide, glibenclamide and glimepiride were carried out. The experiments were carried out at different drug concentrations and different current densities using cathodal iontophoresis. Diffusion cell for iontophoretic permeation study was modified according to Glikfield Design. Pig skin was used for in vitro permeation study and for the in-vivo study New Zealand rabbits were used. At all concentration level iontophoresis showed enhanced permeation rate compared to passive controls. Iontophoretic transports of selected drugs were found to be increased with the current densities. Results showed that target permeation rate for selected drugs could be achieved with the aid of iontophoresis by increasing the area in an appreciable range.

Keywords: transdermal, iontophoresis, pig skin, rabbits, glipizide, glibeclamide

Procedia PDF Downloads 384

Authors: Pugazhenthan Thangaraju

Abstract:

Background: Since the pandemic began, more than millions of people have become infected with COVID-19. Globally, researchers are working for safe and effective treatments for this disease. Remdesivir is a drug that has been approved for the treatment of COVID-19. Many aspects are still being considered that may influence the future use of remdesivir. Aim: To assess the safety and efficacy of Remdesivir in hospitalized adult patients diagnosed with moderate and severe COVID-19. Methods: It was a record-based retrospective cohort study conducted between April 1st, 2020 and June 30th, 2021 at the tertiary care teaching hospital All India Institutes of Medical Sciences (AIIMS), Raipur Results: There were a total of 10,559 medical records of COVID-19 patients of which 1034 records were included in this study. Overall, irrespective of the survival status, there was statistical significant difference observed between the WHO score at the time of admission and discharge. Clinical improvement among the survivors was found to be statistically significant. Conclusion: Remdesivir's potential efficacy against coronaviruses has so far been limited to in vitro studies and animal models. However, information about COVID-19 is rapidly expanding. Several clinical trials for the treatment of COVID-19 with remdesivir are now underway. However, the findings of this study support remdesivir as a promising agent in the fight against SARS-CoV-2.

Keywords: Remdesivir, COVID-19, SARS-CoV-2, antiviral, RNA-dependent RNA polymerase, viral pneumonia

Procedia PDF Downloads 65

Authors: Vishal Kumar Gupta

Abstract:

Purpose: This study has been undertaken to develop novel pH sensitive interpenetrating network hydrogel beads. Methods: The pH sensitive PAAM-g-Guar gum copolymer was synthesized by free radical polymerization followed by alkaline hydrolysis. Beads of guar gum-grafted-polyacrylamide and sodium Carboxy methyl cellulose (Na CMC) loaded with Pantoprazole sodium were prepared and evaluated for pH sensitivity, swelling properties, drug entrapment efficiency and in vitro drug release characteristics. Seven formulations were prepared for the drug with varying polymer and cross linker concentrations. Results: The grafting and alkaline hydrolysis reactions were confirmed by FT-IR spectroscopy. Differential scanning calorimetry was carried out to know the compatibility of encapsulated drug with the polymers. Scanning electron microscopic study revealed that the IPN beads were spherical. The entrapment efficiency was found to be in the range of 85-92%. Particle size analysis was carried out by optical microscopy. As the pH of the medium was changed from 1.2 to 7.4, a considerable increase in swelling was observed for all beads. Increase in the copolymer concentration showed sustained the drug release up to 12 hrs. Drug release from the beads followed super case II transport mechanism. Conclusion: It was concluded that guar gum-acrylamide beads, cross-linked with aluminum chloride offer an opportunity for controlled drug release of pantoprazole sodium.

Keywords: IPN, hydrogels, DSC, SEM

Procedia PDF Downloads 269

Authors: Ali Fattahi Bafghi, Abolghasem Siyadatpanah, Farzaneh Mirzaei, Fahimeh Pournasir, Roghayeh Norouzi, Maria De Lourdes Pereira

Abstract:

Leishmaniasis caused by Leishmania major is one of the main infectious diseases that affect populations in developing countries around the world. We assessed the effectiveness of berberine chloride nano-liposome (BcNLs) against L. major promastigotes in vitro. Nano-liposomal berberine chloride was prepared using the thin-film hydration method and characterized based on encapsulation efficiency, size, and zeta potential. Anti-Leishmania effect of different concentrations (0.05-60 µg/ml) of BcNLs as studied in L. major [MRHO/IR/75/ER] at 24, 48, and 72 h using the hemocytometer technique. Berberine chloride was successfully loaded into nano-liposomes with an encapsulation efficiency of 85.54%. The surface charge of nanoparticles is neutral, and the morphology of nano-liposomal berberine chloride is spherical without any agglomeration. Cell viability assay was performed on the HFF cell line to show the biocompatibility of liposome nanoparticles. IC50 of BcNPs at 24, 48, and 72 h against L. major were found to be 7.6, 5.96, and 3.19 µg/ml, respectively. BcNLs showed a significant anti-Leishmania effect and induced a better and more tangible effect on the survival of L. major promastigotes and could be suitable candidates for further investigation. The results showed that the BcNLs agent is effective against L. major promastigotes and may be a promising alternative to current treatments.

Keywords: Leishmania major, berberine chloride, nano-liposomes, cutaneous leishmaniasis

Procedia PDF Downloads 151

Authors: E. Roshan Ara Begum, K. Bhavani, K. Subachitra, C. Kirthika, R. Shenbagarathai

Abstract:

In recent years, there has been a growing concern for the production of chitosan blend nanofibrous scaffold for its favorable physicochemical properties which mimic the native extracellular matrix (ECM) both morphologically and chemically. Therefore, this study focused on production of β-chitosan(β-Cts) and Poly(vinyl alcohol)(PVA) blend nanofibrous scaffold by electrospinning. β-Cts was extracted from the squid pen waste of locally available squid variety Loligo duvauceli (Indian Squid). To the best of our knowledge, there are no reports on nanofibers preparation from the extracted β-Cts. Both the β-Cts and PVA polymers were mixed in two different proportions (30:70 and 40:60 respectively. The electrospun nanofibrous scaffolds were characterized by SEM, swelling property, in vitro enzymatic degradation, and hemo, biocompatibility properties. β-Cts/PVA nanofibers scaffolds had an average fiber diameter of 120 to 550nm.Among the two different β-Cts/PVA blends nanofibers the β-Cts/PVA (40:60) blend fibers demonstrated favourable tissue engineering properties. The β-Cts/PVA (40:60) blend nanofibers exhibited a swelling ratio of 36 ± 2.5% with mass loss percentage of 20 ± 2.71% after 4 weeks of degradation. It has exhibited good hemocompatible properties. HEK-293(Human Embryonic Kidney) cells lines were able to adhere and proliferate well in the β-Cts/PVA blends nanofibers. All these results indicated that electrospun β-Cts/PVA blends nanofibers are a suitable scaffold to be used for tissue engineering purposes.

Keywords: β-chitosan, electrospinning, nanofibers, poly(vinyl alcohol) (PVA)

Procedia PDF Downloads 235

Authors: Yunhui Chen, Damon Kent, Matthew Dargusch

Abstract:

Synthetic scaffolds are a highly promising new approach to replace both autografts and allografts to repair and remodel damaged bone tissue. Biocompatible porous titanium scaffold was manufactured through a powder metallurgy approach. Magnesium powder was used as space holder material which was compacted with titanium powder and removed during sintering. Evaluation of the porosity and mechanical properties showed a high level of compatibility with human bone. Interconnectivity between pores is higher than 95% for porosity as low as 30%. The elastic moduli are 39 GPa, 16 GPa and 9 GPa for 30%, 40% and 50% porosity samples which match well to that of natural bone (4-30 GPa). The yield strengths for 30% and 40% porosity samples of 315 MPa and 175 MPa are superior to that of human bone (130-180 MPa). In-vitro cell culture tests on the scaffold samples using Human Mesenchymal Stem Cells (hMSCs) demonstrated their biocompatibility and indicated osseointegration potential. The scaffolds allowed cells to adhere and spread both on the surface and inside the pore structures. With increasing levels of porosity/interconnectivity, improved cell proliferation is obtained within the pores. It is concluded that samples with 30% porosity exhibit the best biocompatibility. The results suggest that porous titanium scaffolds generated using this manufacturing route have excellent potential for hard tissue engineering applications.

Keywords: scaffolds, MG-63 cell culture, titanium, space holder

Procedia PDF Downloads 235

Authors: Ravi R. Patel, Vasudev R. Thakkar

Abstract:

Use of plant growth-promoting rhizobacteria (PGPR) to increase the production and decrees disease occurrence is a recent method in agriculture. An RA2 rhizospheric culture was isolated from peanut rhizosphere from Junagadh region of Gujarat, India and showed different direct and indirect plant growth promoting activity like indole acetic acid, gibberellic acid, siderophore, hydrogen cyanide, Ammonia and (1-Aminocyclopropane-1-Carboxylate) deaminase production, N2 fixation, phosphate and potassium solubilization in vitro. RA2 was able to protect peanut germinating seedling from A. niger infection and reduce collar rot disease incidence 60-35% to 72-41% and increase germination percentage from 70-82% to 75-97% in two varieties GG20 and GG2 of peanut. RA2 was found to induce resistance in A. hypogaea L. seedlings via induction of different defense-related enzymes like phenylalanine ammonia lyase, peroxidase, polyphenol oxidase, lipoxygenase and pathogenesis related protein like chitinase, ß – 1,3- glucanase. Jasmonic acid one of the major signaling molecules of inducing systemic resistance was also found to induced due to RA2 treatments. RA2 bacterium was also promoting peanut growth and reduce A. niger infection in pot studies. 16S rDNA sequence of RA2 showed 99 % homology to Azotobacter species.

Keywords: plant growth promoting rhizobacteria, peanut, aspergillus niger, induce systemic resistance

Procedia PDF Downloads 242

Authors: Shokooh Moghadam, Mohammad Taghi Khorasani, Sajjad Seifi Mofarah, M. Daliri

Abstract:

Through the recent two decades, the use of magnetic-property materials with the aim of target cell’s separation and eventually cancer treatment has incredibly increased. Numerous factors can alter the efficacy of this method on curing. In this project, the effect of magnetic field on adhesion of PDL and L929 cells on nanocomposite of iron oxide/PHB with different density of iron oxides (1%, 2.5%, 5%) has been studied. The nanocamposite mentioned includes a polymeric film of poly hydroxyl butyrate and γ-Fe2O3 particles with the average size of 25 nanometer dispersed in it and during this process, poly vinyl alcohol with 98% hydrolyzed and 78000 molecular weight was used as an emulsion to achieve uniform distribution. In order to get the homogenous film, the solution of PHB and iron oxide nanoparticles were put in a dry freezer and in liquid nitrogen, which resulted in a uniform porous scaffold and for removing porosities a 100◦C press was used. After the synthesis of a desirable nanocomposite film, many different tests were performed, First, the particles size and their distribution in the film were evaluated by transmission electron microscopy (TEM) and even FTIR analysis and DMTA test were run in order to observe and accredit the chemical connections and mechanical properties of nanocomposites respectively. By comparing the graphs of case and control samples, it was established that adding nano particles caused an increase in crystallization temperature and the more density of γ-Fe2O3 lead to more Tg (glass temperature). Furthermore, its dispersion range and dumping property of samples were raised up. Moreover, the toxicity, morphologic changes and adhesion of fibroblast and cancer cells were evaluated by a variety of tests. All samples were grown in different density and in contact with cells for 24 and 48 hours within the magnetic fields of 2×10^-3 Tesla. After 48 hours, the samples were photographed with an optic and SEM and no sign of toxicity was traced. The number of cancer cells in the case of sample group was fairly more than the control group. However, there are many gaps and unclear aspects to use magnetic field and their effects in cancer and all diseases treatments yet to be discovered, not to neglect that there have been prominent step on this way in these recent years and we hope this project can be at least a minimum movement in this issue.

Keywords: nanocomposite, cell attachment, magnetic field, cytotoxicity

Procedia PDF Downloads 259

Authors: Kang-Hyun Leem, Myung-Gyou Kim, Hye Kyung Kim

Abstract:

Velvet antler (VA), the immature antlers of male deer, is traditionally used for thousands of years in Asian countries, such as Korea, China, Taiwan, and Mongolia. It has been considered to improve immune system and physical strength. The goal of this study was to investigate the immunomodulatory effect of deer antler velvet using in vitro system. In the first step, the effects of VA (70% ethanol extract) on the proliferation of splenocytes, bone marrow cell, and macrophages were determined. Next, the effect of VA on the production of nitric oxide and phagocytic activity in macrophage were measured. The results showed that VA treatment increased concanavalin-A stimulated splenocyte, bone marrow cells, and macrophage proliferation in a dose dependent manner. VA at 50 and 100 ug/mL concentrations significantly enhanced the concanavalin-A stimulated splenocyte proliferation by 8.8% and 18.5%, respectively. The proliferation of bone marrow cells, isolated from 5wk-old ICR mice, were increased by 25.2% and 46.5% by 50 and 100 ug/mL VA treatment. RAW 264.7 cell proliferation reached peak value at 50 ug/mL of VA treatment exhibiting 108% of the basal value. Nitric oxide production by RAW 264.7 macrophage cells was slightly reduced by VA treatment but was not statistically significant. Moreover, the phagocytic activity of macrophages was enhanced by VA treatment. These results indicate that VA is effective in immune system.

Keywords: deer antler, splenocyte, bone marrow cells, macrophage proliferation, phagocytosis

Procedia PDF Downloads 272

Authors: Yantao Li, Jun Zheng

Abstract:

Food poison caused by consumption of contaminated food, especially seafood, is one of most serious public health threats worldwide. Vibrio parahaemolyticus is emerging bacterial pathogen and the leading cause of human gastroenteritis associated with food poison, especially in the southern coastal region of China. To successfully cause disease in host, bacterial pathogens need to overcome the host-derived stresses encountered during infection. One of the toxic chemical species elaborated by the host is nitric oxide (NO). NO is generated by acidified nitrite in the stomach and by enzymes of the inducible NO synthase (iNOS) in the host cell, and is toxic to bacteria. Bacterial pathogens have evolved some mechanisms to battle with this toxic stress. Such mechanisms include genes to sense NO produced from immune system and activate others to detoxify NO toxicity, and genes to repair the damage caused by toxic reactive nitrogen species (RNS) generated during NO toxic stress. However, little is known about the NO resistance in V. parahaemolyticus. In this study, a transposon coupled with next generation sequencing (Tn-seq) technology will be utilized to identify genes for NO resistance in V. parahaemolyticus. Our strategy will include construction the saturating transposon insertion library, transposon library challenging with NO, next generation sequencing (NGS), bioinformatics analysis and verification of the identified genes in vitro and in vivo.

Keywords: vibrio parahaemolyticus, nitric oxide, tn-seq, virulence

Procedia PDF Downloads 264

Authors: Salem Abdalla

Abstract:

Objective: Dipyrone is a widely used, well tolerated analgesic drug which, however, is compromised by agranulocytosis as an adverse effect. Subsequent to no enzymatic hydrolysis, the primary metabolic step is N-demethylation of 4-methylaminoantipyrine (4-MAA) to 4-aminoantipyrine (4-AA). The aim of the present study was to identify the cytochrome P-450 enzyme (CYP) mediating this reaction. Methods: We identified the relevant CYP using virus expressed isolated rat liver microsomes with chemical inhibition studies. The substrate of 4-methylaminantipyrine was employed at six different concentrations (25, 50, 100, 400, 800, and 1200 µmol/l) with varying concentrations of selective inhibitors of CYP1A2 (furafylline, fluvoxamine), CYP3A4 (ketoconazole), CYP2A6 (coumarin), CYP2D6 (quinidine), CYP2C19 (omeprazole, fluvoxamine, tranylcypromine), CYP2C9 (sulfaphenazole), and CYP1A1 (alpha-naphthoflavone). 4-MAA and 4-AA were analyzed by HPLC, and enzyme kinetic parameters (Km and Vmax) were determined by regression (Sigma plot 9.0). Results: The N-demethylation of 4-MAA by microsomes prepared from baculovirus-expressing human CYP was pronounced with CYP2C19. Intrinsic clearances of the most active enzymes were 0.092, 0.027, and 0.026 for the CYP enzymes 2C19, 2D6, and 1A2, respectively. Metabolism by rat liver microsomes was strongly inhibited by omeprazole (IC50 of 0.05). Conclusion: The enzyme CYP2C19 apparently has an important role in N-demethylation of 4-methylaminoantipyrine which should be further analyzed in clinical studies and which may also be interesting concerning the agranulocytosis.

Keywords: dipyrone, 4-methylaminoantipyrine (4-MAA), 4- aminoantipyrine (4-AA), metabolism, human CYP2C19

Procedia PDF Downloads 238

Authors: Amine Ez-Zoubi, Abdellah Farah

Abstract:

Because of their effectiveness and environmental safety, many essential oils have been investigated as biopesticides. Nevertheless, the encapsulation process is necessary to improve its physical, chemical, and biological properties. Therefore, the purpose of this paper was to study the physicochemical characteristics, and antifungal activity of the Artemisia Herba-Alba essential oil (HAEO) encapsulated in succinic acid modified β-CD (SACD). A yellowish oil was obtained from plant A. Herba-Alba using hydrodistillation and GC-MS was used to identify the chemical composition, in which α-Thujone (65.0%) was the main component in HAEO. The succinic acid has been esterified via the hydroxyl groups in β-CD to produce SACD. In addition, the inclusion complex formation of HAEO and SACD was generated according to the co-precipitation method and was analyzed by several techniques. The antifungal activity in vitro was examined against Botrytis cinerea by direct contact with a potato dextrose agar culture medium. At a 0.1 % concentration, the HAEO in encapsulated form showed higher potential for the control of B. cinerea when compared to the EO in free form (38.34 to 12%). Thus, these results produced evidence that the encapsulation of EOs in SACD can be useful for the development of B.cinerea inhibitors and a promising alternative biopesticide.

Keywords: Artemisia Herba-Alba essential oil, succinic acid modified β-cyclodextrin, inclusion complex, co-precipitation, Botrytis cinerea, direct contact

Procedia PDF Downloads 87

Authors: Da-Ming Huang, Ya-Ting Tsai, Shyh-Hau Wang

Abstract:

Ultrasound signals backscattered from the soft tissues are mainly depending on the size, density, distribution, and other elastic properties of scatterers in the interrogated sample volume. The quantitative analysis of ultrasonic backscattering is frequently implemented using the statistical approach due to that of backscattering signals tends to be with the nature of the random variable. Thus, the statistical analysis, such as Nakagami statistics, has been applied to characterize the density and distribution of scatterers of a sample. Yet, the accuracy of statistical analysis could be readily affected by the receiving signals associated with the nature of incident ultrasound wave and acoustical properties of samples. Thus, in the present study, efforts were made to explore such effects as the ultrasound operational modes and attenuation of biological tissue on the estimation of corresponding Nakagami statistical parameter (m parameter). In vitro measurements were performed from healthy and pathological fibrosis porcine livers using different single-element ultrasound transducers and duty cycles of incident tone burst ranging respectively from 3.5 to 7.5 MHz and 10 to 50%. Results demonstrated that the estimated m parameter tends to be sensitively affected by the use of ultrasound operational modes as well as the tissue attenuation. The healthy and pathological tissues may be characterized quantitatively by m parameter under fixed measurement conditions and proper calibration.

Keywords: ultrasound backscattering, statistical analysis, operational mode, attenuation

Procedia PDF Downloads 323

Authors: Zerihun Tesfaye Nigusse

Abstract:

Objective: This study evaluates the antileishmanial activity of the methanolic extract of Aloe otallensis (A. otallensis) on the promastigote stage of Leishmaniadonovani (L. donovani) as compared to standard drugs and to screen its phytochemical constituents. Methods: Phytochemical screening was done by using the method mentioned by Evans and Trease on methanolic extract of the exudates of Aloe otallensis leaves. The extract was also evaluated for in vitro antileishmanial activity against L. donavani, which is found in the Parasitology Unit of Black Lion Hospital. The result was compared to standard drugs of sodium stibogluconate, milfostin and paramomycin. Results: The extract has good antileishmanial activity with an IC50 of 0.123 0 μg/mL on L. donovani (AM 563). The experimental data showed that relatively, it had better activity than paramomycin and milfostin but less activity than sodium stibogluconate. The data analyses were done by GraphPad Prism version 5 software after it was read by an ELISA reader at the wavelength of 650 nm. The phytochemical screening of the exudates of A. otallensis showed the presence of phenol, alkaloid and saponin. Conclusions: The methanol extract of the exudates of A.otallensishas a good anti- leishmaniasis activity and this may be attributed to phenol, alkaloid and saponin present in the plant. But it needs further analysis for the conformation of which constituent presents in high concentration to know which one has the strongest effect.

Keywords: anti leshimaniasis, aloe otallensis, leshimania ethiopica, IC50

Procedia PDF Downloads 38

Authors: Berat Krasniqi, Shpëtim Thaçi, Miribane Dërmaku-Sopjani, Sokol Abazi, Mentor Sopjani

Abstract:

The Mediterranean native plant, Sideritis raeseri Boiss. & Heldr. (Lamiaceae), also known as "mountain tea," has a long history of use in traditional medicine. The effects of an ethanol extract of Sideritis raeseri (SR) on uterus smooth muscle activity are evaluated in this study, and the underlying mechanism is identified. S. raeseri extract (SRE) was made from air-dried components of the SR shoot system. At 37°C, the SRE (0.5-2 mg/mL) was tested on isolated rabbit uterus rings that were suspended in a Krebs solution-filled organ bath and bubbled with a mixture of 95% O₂ and 5% CO₂. The SRE alone relaxed the muscle contraction in a concentration-dependent manner in uterine rings in in vitro tests. SRE also decreased Ca²⁺-induced contractions in the uterus by a large amount when the uterus was depolarized with carbachol (CCh, 1µM), K⁺ (80 mM), or contracted by oxytocin (5 nM). The potential involvement of NO-dependent or independent cGMP mechanisms in the uterine actions of SR was investigated. For this purpose, L-NAME (NO synthase inhibitor, 100 M) or bradykinin (NO synthase stimulator, 100 nM), or indomethacin (cyclooxygenase inhibitor, 10µM) decreased the impact of SRE. These results suggest that NO-dependent signaling is involved in SRE's mediated uterine relaxant effect. Data suggests that SRE could be a powerful tocolytic agent that reduces uterine activity and could be used to treat a number of uterine conditions.

Keywords: Sideritis raeseri, uterus, alternative medicine, intracellular mechanisms

Procedia PDF Downloads 116

Authors: Astha Saxena

Abstract:

The present study is based in Indian subcontinent and aims at exploring students’ conceptions about ethical issues related to Biotechnology at both high school and undergraduate level. The data collection methods involved taking classroom notes, recording students’ observations and arguments, and focussed group discussions with students. The data was analysed using classroom discourse analysis and interpretive approaches. The findings depicted different aspects of students’ thinking, meaning making and ethical understanding with respect to complex bioethical issues such as genetically modified crops, in-vitro fertilization (IVF), human genomic project, cloning, etc., at high school as well as undergraduate level. The paper offers a comparative account of students’ arguments with respect to ethical issues in biotechnology at the high school & undergraduate level, where it shows a clear gradation in their ethical understanding from high school to undergraduate level, which can be attributed to their enhanced subject-matter knowledge. The nature of students’ arguments reveal that there is more reliance on the utilitarian aspect of these biotechnologies as against a holistic understanding about a particular bioethical issue. This study has implications for science teachers to delve into students’ thinking and notions about ethical issues in biotechnology and accordingly design appropriate pedagogical approaches.

Keywords: ethical issues, biotechnology, ethical understanding, argument, ethical reasoning, pedagogy

Procedia PDF Downloads 80

Authors: Dagim Ali Hussen, Adnan A. Bekhit, Ariaya Hymete

Abstract:

In this study, several pyridine derivatives were synthesized and evaluated for their in vitro antimicrobial activity against gram-positive bacteria (S. aureus and B. Cereus), gram-negative bacteria (P. aeruginosa and E. coli) and fungus (C. albican and A niger). The intermediate chalcone derivative 2a,b was synthesized by condensation of pyrazole aldehydes 1a,b with acetophenone in alcoholic KOH. Cyclization of 2a,b with ethyl cyanoacetate ad ammonium acetate resulted in pyridine carbonitrile derivatives 3a,b. Furthermore, condensation of pyridine-4-carboxaldeyhe with different amino-derivatives gave rise to pyridine derivatives 5a,b, 6a,b. The oxadiazole derivative 7a was prepared by cyclization of 6a with acetic anhydride. Characterization of the synthesized compound was performed using IR, 1H NMR, 13C NMR spectra and elemental microanalyses. The antimicrobial results revealed that compounds 5a, 6b and 7a exhibited half fold antibacterial activity compared to ampicillin, against B. cereus. On the other hand, compound 3b showed an equivalent activity compared to miconazole against candida albican (CANDAL 03) and to clotrimazole against the clinical isolate candida albican 6647. Moreover, this compound 3b was further tested for its acute toxicity profile. The results showed that oral LD50 is more that 300 mg/kg and parentral LD50 is more than 100 mg/kg. Compound 3b is a good candidate for antifungal agent with good toxicity profile, and deserves more chemical derivatization and clinical study.

Keywords: antifungal, antimicrobial, Candida albican, pyridine

Procedia PDF Downloads 498

Authors: Nattawit Thiapairat

Abstract:

Multiple diseases have been linked to excessive levels of free radicals, which cause tissue or cell damage as a result of oxidative stress. Many plants are sources of high antioxidant activity. Derris scandens has a high amount of phenolic and flavonoid contents which demonstrated good biological activities. This study focused on the antioxidant activity of polyphenols extracted from D. scandens. This study performs total flavonoids content and various antioxidant assays, which were 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging capacity assays. The total flavonoid content of D. scandens extract was determined and expressed as quercetin equivalents (QE)/g measured by the aluminum chloride colorimetric method. The antioxidant activity of D. scandens extract was also determined by DPPH and ABTS assays. In the DPPH assay, vitamin C was used as a positive control, whereas Trolox was used as a positive control in the ABTS assay. The half-maximal inhibitory concentration (IC50) values for D. scandens extract from DPPH and ABTS assays were 41.79 μg/mL ± 0.783 and 29.42 μg/mL ± 0.890, respectively, in the DPPH assay. To conclude, D. scandens extract consists of a high amount of total phenolic content, which exhibits a significant antioxidant activity. However, further investigation regarding antioxidant activity such as SOD, ROS, and RNS scavenging assays and in vivo experiments should be performed.

Keywords: ABTS assay, antioxidant activity, Derris scandens, DPPH assays, total flavonoid content

Procedia PDF Downloads 213

Authors: Ahmad Shah Farhat

Abstract:

Background: Prenatal asphyxia or birth asphyxia is the medical situation resulting from a newborn infant that lasts long enough during the birth process to cause physical harm, usually to the brain. Human umbilical cord blood (UCB) is a well-established source of hematopoietic stem/progenitor cells (HSPCs) for allogeneic stem cell transplantation. These can be used clinically to care for children with malignant diseases. Low O2 can cause in proliferation and differentiation of stem cells. Method: the cord blood of 11 infants with 3-5 Apgar scores or need to cardiac pulmonary Resuscitation as an asphyxia group and ten normal infants with more than 8 Apgar scores as the normal group was collected, and after isolating hematopoietic stem cells, the cells were cultured in enriched media for 14 days to compare the numbers of colonies by microscope. Results: There was a significant difference in the number of RBC precursor colonies (red colonies) in cultured media with 107 cord blood hematopoietic stem cells of infants who were exposed to hypoxemia in two wells of palate. There was not a significant difference in the number of white cell colonies in the two groups in the two wells of the plate. Conclusion: Hypoxia in the perinatal period can cause the increase of hematopoietic stem cells of cord blood, special red precursor stem cells in vitro, like an increase of red blood cells in the body when exposed to low oxygen conditions. Thus, it will be usable.

Keywords: asphyxia, neonre, stem cell, red cell

Procedia PDF Downloads 77

Authors: Sayaka Ono, Ryutaro Imai, Tomoko Ehara, Tetsuya Matsumoto, Hajime Matsumura

Abstract:

Background: Wound pH affects a number of important factors in wound healing. We previously measured the pH value of the exudates collected from second-degree burns and found that the increase in pH was observed in the burn wounds in which colonized by Staphylococcus spp., and the increase in pH was evident prior to the clinical findings of local infection. To investigate the relationship between the changes of pH value and bacterial growth, we performed in vitro study using Pseudomonas aeruginosa and liquid medium as a locally infected wound equivalent model. Methods: Pseudomonas aeruginosa standard strain (ATCCR 10145TM) was cultured at 37 °C environment in Luria Broth Miller medium. The absorbance rate which means the amount of bacteria was measured by a microplate reader 2300EnSpireTM). The pH was measured using pH-indicator strips (MColorpHastTM). The statistical analysis was performed using the product-moment correlation coefficient of Pearson's. Results: The absorbance rate and pH value were increased along with culture period. There was a positive correlation between pH value and absorbance rate (n = 27, Pearson's r = 0.985). Moreover, there was a positive correlation between pH value and the culture period (n = 18, Pearson's r = 0.901). The bacteria was well growth in the media from pH 6.6 to pH 8.0 and the pH of culture media converged at 8 -9 along with the bacterial growth. Conclusion: From these results, we conclude that pH value of the wound is correlated with the number of viable bacteria and bacterial growth periods.

Keywords: colonization, potential of hydrogen, Pseudomonas aeruginosa, wound

Procedia PDF Downloads 279

Authors: Orapan Paecharoenchai, Tanasait Ngawhirunpat, Theerasak Rojanarata, Auayporn Apirakaramwong, Praneet Opanasopit

Abstract:

Cationic niosomes formulated with Span20, cholesterol and novel synthesized spermine-cationic lipids (2-hydrocarbon tail and 4- hydrocarbon tail) in a molar ratio of 2.5:2.5:1 can mediate high gene transfection in vitro. However, the uptake mechanisms of these systems are not well clarified. In the present study, effect of endocytic inhibitors on the transfection efficiency of niosomes/DNA complexes was determined on a human cervical carcinoma cell line (HeLa cells) using the inhibitors of macropinocytosis (wortmannin), clathrin- and caveolae-mediated endocytosis (methyl-β-cyclodextrin), clathrin-mediated endocytosis (chlorpromazine), caveolae-mediated endocytosis (genistein and filipin), cytosolic transfer (ammonium chloride) and microtubules polymerization (nocodazole). The transfection of niosomes with 2-hydrocarbon tail lipid was blocked by nocodazole, genistein, ammonium chloride and filipin, respectively, whereas, the transfection of niosomes with 4-hydrocarbon tail lipid was blocked by nocodazole, genistein, ammonium chloride, methyl-β-cyclodextrin and filipin, respectively. It can be concluded that these niosomes/DNA complexes were internalized predominantly by endocytosis via clathrin and caveolae-independent pathway.

Keywords: cellular internalization, cationic niosomes, gene carriers, spermine-cationic lipids

Procedia PDF Downloads 456

Authors: Li-Yu Lee, Yu-Shuan Chen, Jun Sheng Wang, I-Ming Chu

Abstract:

Microneedle patch is a painless transdermal drug delivery method, It could solve some problems in traditional drug delivery such as digestive system causing drug metabolism and subcutaneous injection causing some side effects. Coating drug on or loading drug in microneedle can carry active ingredient through stratum corneum, also can control dose well when microneedle patch apply on localized topical area. We used hyaluronic acid to fabricate dissolvable microneedle patch and encapsulated minoxidil into microneedles. Minoxdil is a drug for exterior use that can be used to treat Androgenetic alopecia, but related commercial products have some shortcomings, for example, propylene glycol which is used to soften stratum corneum cause skin allergic reaction, comparing chemical promotion, microneedle patch provide physical way to make drugs through nature barrier of skin. In this research, we designed a two-step process to fabricate microneedle patch, that can effectively reduce drug waste, and gentle production process could maintain drug activity well. We also do in vitro test on cadaver to make sure patch has enough mechanical strength to penetrate stratum corneum. In the release test and animal test, we found microneedle patch has higher delivery efficiency than tradition way. In this study, we may determine that germinal MNs patch is a potential commodity.

Keywords: dissolving microneedles, androgenetic alopecia, minoxidil, transdermal drug delivery

Procedia PDF Downloads 279

Authors: Katharigatta N. Venugopala, Melendhran Pillay, Bander E. Al-Dhubiab

Abstract:

Tuberculosis (TB) infection is caused mainly by Mycobacterium tuberculosis (MTB) and it is one of the most threatening and wide spread infectious diseases in the world. Benzothiazole derivatives are found to have diverse chemical reactivity and broad spectrum of pharmacological activity. Some of the important pharmacological activities shown by the benzothiazole analogues are antitumor, anti-inflammatory, antimicrobial, anti-tubercular, anti-leishmanial, anticonvulsant and anti-HIV properties. Keeping all these facts in mind in the present investigation it was envisaged to synthesize a series of novel {2-(benzo[d]-thiazol-2-yl-methoxy)-substitutedaryl}-(substitutedaryl)-methanones (4a-f) and characterize by IR, NMR (1H and 13C), HRMS and single crystal x-ray studies. The title compounds are investigated for in vitro anti-tubercular activity against two TB strains such as H37Rv (ATCC 25177) and MDR-MTB (multi drug resistant MTB resistant to Isoniazid, Rifampicin and Ethambutol) by agar diffusion method. Among the synthesized compounds in the series, test compound {2-(benzo[d]thiazol-2-yl-methoxy)-5-fluorophenyl}-(4-chlorophenyl)-methanone (2c) was found to exhibit significant activity with MICs of 1 µg/mL and 2 µg/mL against H37Rv and MDR-MTB, respectively when compared to standard drugs. Single crystal x-ray studies was used to study intra and intermolecular interactions, including polymorphism behavior of the test compounds, but none of the compounds exhibited polymorphism behavior.

Keywords: benzothiazole analogues, characterization, crystallography, anti-TB activity

Procedia PDF Downloads 281

Authors: Patcharakamon Nooeaid, Piyachat Chuysrinuan

Abstract:

Electrospun natural polymers-based nanofibers are one of the most interesting materials used in tissue engineering and drug delivery applications. Bead-on-string nanofibers have gained considerable interest for sustained drug release. Vancomycin was used as the model drug and sodium alginate (SA)/soy protein isolated (SPI) as the polymer blend to fabricate the bead-on-string nanofibers by aqueous-based electrospinning. The bead-on-string SA/SPI nanofibers were successfully fabricated by the addition of poly(ethylene oxide) (PEO) as a co-blending polymer. SA-PEO with mass ratio of 70/30 showed the best spinnability with continuous nanofibers without the occurrence of beads. Bead structure formed with the addition of SPI and bead number increased with increasing SPI content. The electrospinning of 80/20 SA-PEO/SPI was obtained as a great promising bead-on-string nanofibers for drug loading, while the solution of 50/50 was not able to obtain continuous fibers. In vitro release tests showed that a more sustainable release profile up to 14 days with less initial burst release on day 1 could be obtained from the bead-on-string fibers than from smooth fibers with uniform diameter. In addition, vancomycin-loaded beaded fibers inhibited the growth of Staphylococcus aureus (S. aureus) bacteria. Therefore, the SA-PEO/SPI nanofibers showed the potential to be used as biomaterials for tissue engineering and drug delivery.

Keywords: bead-on-string fibers, electrospinning, drug delivery, tissue engineering

Procedia PDF Downloads 334

Authors: Dana A. Thal, Heike Kahlert, Fritz Scholz

Abstract:

Thiol molecules are known to easily form self-assembled monolayers (SAM) on Au surfaces. Depending on the thiol’s structure, surface modifications via SAM can be used for electrode sensor development. In the presented work, 1-decanethiol coated polycrystalline Au electrodes were applied to indirectly assess the radical scavenging potential of plant compounds and extracts. Different plant compounds with reported antioxidant properties as well as an extract from the plant Gynostemma pentaphyllum were tested for their effectiveness to prevent SAM degradation on the sensor electrodes via photolytically generated radicals in aqueous media. The SAM degradation was monitored over time by differential pulse voltammetry (DPV) measurements. The results were compared to established antioxidant assays. The obtained data showed an exposure time and concentration dependent degradation process of the SAM at the electrode’s surfaces. The tested substances differed in their capacity to prevent SAM degradation. Calculated radical scavenging activities of the tested plant compounds were different for different assays. The presented method poses a simple system for radical scavenging evaluation and, considering the importance of the test system in antioxidant activity evaluation, might be taken as a bridging tool between in-vivo and in-vitro antioxidant assay in order to obtain more biologically relevant results in antioxidant research.

Keywords: alkanethiol SAM, plant antioxidant, polycrystalline Au, radical scavenger

Procedia PDF Downloads 298