Search results for: In vitro

Authors: Krishna Poudel, Tahar Katuwal, Sujan Karki

Abstract:

A rapid propagation and acclimatization method of large cardamom was optimized in this study. Sprouted rhizome buds were collected. The excised rhizome bud explants were cultured on semi solid culture media. The explants were cultured on Murashige and Skoog’s (MS) medium supplemented with different concentration and combinations of BAP (6-Benzyl-amino-purine) and IBA (Indole-3-butyric acid) for shoot and root induction. Explants cultured on MS basal medium supplemented with 1.0 mg/l BAP + 0.5 gm/l IBA showed the highest rate of shoot multiplication. In vitro shoots were rooted on to the half-strength MS basal media supplemented with 0.5 mg/l IBA. Rooted shoots were transplanted in the screen house for hardening process. These hardened plants were subsequently shifted into the netted nursery for further multiplication process.

Keywords: concentration, explants, hardening, rhizome

Procedia PDF Downloads 208

Authors: Eun Jeong Yu, Inn Soo Kang, Tae Ki Yoon, Mi Kyoung Koong

Abstract:

Advanced female age is associated with higher cycle cancelation rates, lower clinical pregnancy rate, increased miscarriage and aneuploidy rates in IVF (In Vitro Fertilization) cycles. This retrospective cohort study was conducted at a Cha Fertility Center, Seoul Station. All fresh non-donor IVF cycles performed in women aged 40 years and above from January 2016 to December 2016 were reviewed. Donor/recipient treatment, PGD/PGS (Preimplantation Genetic Diagnosis/ Preimplantation Genetic Screening) were excluded from analysis. Of the 1,166 cycles from 753 women who completed ovulation induction, 1,047 were appropriate for the evaluation according to inclusion and exclusion criterion. IVF cycles were categorized according to age and grouped into the following 1-year age groups: 40, 41, 42, 43, 44, 45 and > 46. The mean age of patients was 42.4 ± 1.8 years. The median AMH (Anti-Mullerian Hormone) level was 1.2 ± 1.5 ng/mL. The mean number of retrieved oocytes was 4.9 ± 4.3. The clinical pregnancy rate and live birth rate in women > 40 years significantly decreased with each year of advancing age (p < 0.001). The clinical pregnancy rate decreased from 21% at the age of 40 years to 0% at ages above 45 years. Live birth rate decreased from 12.3% to 0%, respectively. There were no clinical pregnancy outcomes among 95 patients aged above 45 years of age. The overall miscarriage rate was 40.7% (range, 36.7%-70%). The transfer of at least one good quality embryo was associated with about 4-9% increased chance of a clinical pregnancy rate. Therefore, IVF in old age women less than 46 had a reasonable chance for successful pregnancy outcomes especially when good quality embryo is transferred.

Keywords: advanced maternal age, in vitro fertilization, pregnancy rate, live birth rate

Procedia PDF Downloads 113

Authors: Deepika Saini, Sandeep Jain, Ajay Kumar

Abstract:

The quinoline scaffold is one of the most widely studied in the discovery of derivatives with various heterocyclic moieties due to its potential antimalarial activities. In the present study, a chalcone series of quinoline derivatives clubbed with pyrazole were synthesized to evaluate their antimalarial property by in vitro schizont maturation inhibition assay against both chloroquine sensitive, 3D7 and chloroquine resistant, RKL9 strain of Plasmodium falciparum. Further, top five compounds were studied for in vivo preclinical study for antimalarial potential against P. berghei in Swiss albino mice. To understand the mechanism of synthesized analogues, they were screened computationally by molecular docking techniques. Compounds were docked into the active site of a protein receptor, Plasmodium falciparum Cysteine Protease Falcipain-2. The compounds were successfully synthesized, and structural confirmation was performed by FTIR, 1H-NMR, mass spectrometry and elemental analysis. In vitro study suggested that the compounds 5b, 5g, 5l, 5s and 5u possessed best antimalarial activity and further tested for in vivo screening. Compound 5u (CH₃ on both rings) with EC₅₀ 0.313 & 0.801 µg/ml against CQ-S & CQ-R strains of P. falciparum respectively and 78.01% suppression of parasitemia. The molecular docking studies of the compounds helped in understanding the mechanism of action against falcipain-2. The present study reveals the binding signatures of the synthesized ligands within the active site of the protein, and it explains the results from in vitro study in their EC₅₀ values and percentage parasitemia.

Keywords: antimalarial activity, chalcone, docking, quinoline

Procedia PDF Downloads 377

Authors: P. S. Percin, O. Inanli, S. Karakaya

Abstract:

Bitter melon (Momordica charantia) is a medicinal vegetable, which is used traditionally to remedy diabetes. Bitter melon contains several classes of primary and secondary metabolites. In traditional Turkish medicine bitter melon is used for wound healing and treatment of peptic ulcers. Nowadays, bitter melon is used for the treatment of diabetes and ulcerative colitis in many countries. The main constituents of bitter melon, which are responsible for the anti-diabetic effects, are triterpene, protein, steroid, alkaloid and phenolic compounds. In this study total phenolics, total carotenoids and β-carotene contents of mature and immature bitter melons were determined. In addition, in vitro α-amylase and α-glucosidase activities of mature and immature bitter melons were studied. Total phenolic contents of immature and mature bitter melon were 74 and 123 mg CE/g bitter melon respectively. Although total phenolics of mature bitter melon was higher than that of immature bitter melon, this difference was not found statistically significant (p > 0.05). Carotenoids, a diverse group of more than 600 naturally occurring red, orange and yellow pigments, play important roles in many physiological processes both in plants and humans. The total carotenoid content of mature bitter melon was 4.36 fold higher than the total carotenoid content of immature bitter melon. The compounds that have hypoglycaemic effect of bitter melon are steroidal saponins known as charantin, insulin-like peptides and alkaloids. α-Amylase is one of the main enzymes in human that is responsible for the breakdown of starch to more simple sugars. Therefore, the inhibitors of this enzyme can delay the carbohydrate digestion and reduce the rate of glucose absorption. The immature bitter melon extract showed α-amylase and α-glucosidase inhibitory activities in vitro. α-Amylase inhibitory activity was higher than that of α-glucosidase inhibitory activity when IC50 values were compared. In conclusion, the present results provide evidence that aqueous extract of bitter melon may have an inhibitory effect on carbohydrate breakdown enzymes.

Keywords: bitter melon, in vitro antidiabetic activity, total carotenoids, total phenols

Procedia PDF Downloads 213

Authors: Venkatalakshmi Ranganathan, Ong Hsin Ju, Tan Yinn Ming, Lim Kien Sin, Wong Man Ting, Venkata Srikanth Meka

Abstract:

The mucoadhesive buccal tablet is an oral drug delivery system which attached to the buccal surface for direct drug absorption into the systemic circulation and the unidirectional drug release is ensured by formulating a hydrophobic backing layer. The objective of present study was to formulate mucoadhesive atenolol bilayer buccal tablets by using sodium alginate, hydroxyethyl cellulose, and xanthan gum as mucoadhesive polymer and the technique applied was direct compression method. Ethyl cellulose was used as backing layer of the tablet. FTIR and DSC analysis were carried out to identify the drug polymer interactions. The prepared tablets were evaluated for physicochemical parameters, ex vivo mucoadhesion time and in-vitro drug release. The formulated tablets showed the average surface pH 6-7 which is favourable for oral mucosa. The formulation containing sodium alginate showed more than 90 % of drug release at the end of the 7 hours in vitro dissolution studies. The formulation containing xanthan gum showed more than 8 hours of mucoadhesion time and all formulation exhibited non fickian release kinetics. The present study indicates enormous potential of erodible mucoadhesive buccal tablet containing atenolol for systemic delivery with an added advantage of circumventing the hepatic first pass metabolism.

Keywords: atenolol, mucoadhesion, in vitro drug release, direct compression, ethyl cellulose

Procedia PDF Downloads 591

Authors: Aakanchha Jain, D. V. Kohli

Abstract:

Cubosomes are nanoparticles but instead of the solid particles, cubosomes are self-assembled liquid crystalline particles of certain surfactant with proper ratio of water with a microstructure that provides unique properties of practical interest. Cubosomes encapsulating Fluconazole were prepared by emulsification method and characterized for particle size, entrapment efficiency. The cubosomes prepared were 257.2±2.94 nm in size with drug entrapment efficiency of 66.2±2.69%. The optimized formulation characterized for shape and surface morphology by TEM and SEM analysis. SEM photograph showed the smooth surface of optimized cubosomes and TEM photograph revealed square somewhat circular intact shapes of cubosomes. MIC was determined by XTT based method and antifungal activity was determined in vitro. The cumulative percentage of Fnz from cubosomes permeated via dialysis membrane (MWCO 12-14 KD) showed a percent cumulative drug release of 76.86% while Fnz solution showed release up to 91.04% in 24 hours in PBS (pH 6.5)(p < 0.005).

Keywords: Candids albicans, cubosomes, fluconazole, topical delivery

Procedia PDF Downloads 272

Authors: Mardiati Zain, Jurnida Rahman, Khasrad, Erpomen

Abstract:

The aim of this experiment was to study the use of palm oil by products (oil palm fronds (OPF), palm oil sludge (POS) and palm kernel cake (PKC)), that supplemented with growth factor rumen microbes (Sapindus rarak and Sacharomyces cerevisiae) on digestibility and fermentation in vitro. Oil Palm Fronds was previously treated with 3% urea. The treatments consist of 50% OPF+ 30% POS+ 20% PKC as a control diet (A), B = A + 4% Sapindus rarak, C = A + 0.5 % Sacharomyces cerevisiae and D = A + 4% Sapindus rarak + 0.5% Sacharomyces cerevisiae. Digestibility of DM, OM, ADF, NDF, cellulose and rumen parameters (NH3 and VFA) of all treatments were significantly different (P < 0.05). Fermentation and digestibility treatment A were significantly lower than treatments B, C, and D. The result indicated that supplementation Sapindus rarak and S. cerevisiae were able to improve fermentability and digestibility of palm oil by product.

Keywords: palm oil by product, Sapindus rarak, Sacharomyces rerevisiae, fermentability, OPF ammoniated

Procedia PDF Downloads 656

Authors: Bouchenak Fatima, Lmegharbi Abdelbaki, Houssem Degaichia, Benrebiha Fatima

Abstract:

The essential oil composition of the leaf of Artemisia absinthium from region of Cherchell (The south of Algeria) was investigated by GC, GC-MS. 27 constituents were identified correspond to 84, 63% of the total oil. The major components are Thujone (60, 82%), Chamazulènel (16, 62%), ρ-cymène (4, 29%) and 2-carène (4.25%). The antimicrobial activity of oil was tested in vitro by two methods (agar diffusion and microdilution) on three plant pathogenic fungi. This oil has been tested for antimicrobial activity against three pathogenic fungi (Botrytis cinerea, Fusarium culmorum and Helminthosporium Sp.).The study of activity was evaluated by two methods: Method of diffusion in gelose and the minimum inhibitory concentration MIC. This oil exhibited an interesting antimicrobial activity. A preliminary study showed that this oil presented high toxicity against this fungus. These results, although preliminary show a good antifungal activity, to limit and inhibit stop the development of those pathogen agent.

Keywords: artemisia absinthian, extraction process, chemical study, antifungal activity

Procedia PDF Downloads 438

Authors: Mahendran Ganesan, Sanjeet Kumar Verma, Zafar Iqbal, Ashish Chandran, Zakir Husain, Shama Afroz, Sana Shahid, Laiq Ur Rahman

Abstract:

Highly efficient in vitro regeneration system has been developed for Swertia chirayita (Roxb. ex Fleming) H. Karst, a high prized traditional medicinal plant to treat numerous ailments such as liver disorders, malaria and diabetes and are reported to have a wide spectrum of pharmacological properties. Its medicinal usage is well-documented in Indian pharmaceutical codex, the British and the American pharmacopeias, and in different traditional medicine such as the Ayurveda, Unani and Siddha medical systems. Nodal explants were cultured on MS medium supplemented with various phytohormones for multiple shoot induction. The nodal segments failed to respond in growth regulator free medium. All the concentrations of BAP, Kin and TDZ facilitated shoot bud break and multiple shoot induction. Among the various cytokinins tested, BAP was found to be more effective with respect to initiation and subsequent development of shoots. Of the various concentrations BAP tested, BAP at 4.0 mg/L showed the higher average number of shoot regeneration (10.80 shoots per explant). Kin at 4 mg/L and TDZ at 4 mg/L induced 5.70 and 04.5+0 shoots per explant, respectively. Further increase in concentration did not favour an increase in the number of shoots. However, these shoots failed to elongate further. Hence, addition of GA₃ (1 mg/L) was added to the above medium. This treatment resulted in the elongation of shoots (2.50 cm) and a further increase in the number of microshoots (34.20 shoots/explant). Roots were also induced in the same medium containing BAP (4 mg/L) + GA₃ (1 mg/L) + NAA (0.5 mg/L). In vitro derived plantlets with well-developed roots were transferred to the potting media containing garden soil: sand: vermicompost (2:1:1). Plantlets were covered with a polyethylene bag and irrigated with water. The pots were maintained at 25 ± 2ºC, and then the polyethylene cover was gradually loosened, thus dropping the humidity (65–70%). This procedure subsequently resulted in in vitro hardening of the plantlet.

Keywords: micropropagation, nodal explant, plant growth regulators, Swertia chirayita

Procedia PDF Downloads 86

Authors: Alaa M. M. Badr Eldin, Marwa I. Ezzat, Mohammed S. Sedeek, Manal S. Afifi, Omar M. Sabry

Abstract:

Cytotoxic activity of the total methanol extract against breast cancer cell line MCF-7 was amazing IC50 at 54 ug/ml. 130 polyphenolic compounds were tentatively identified in pomegranate peel (Punica granatum L.) methanol extract using HPLC-MS/MS technique. The antiestrogenic activity of the polyphenolic constituents found in pomegranate extract was confirmed experimentally in-vitro and by the in-silico molecular docking using gallagic acid, ellagic acid, and Punicalagin as these are considered model compounds confirmed in pomegranate peel extract. The methanolic extract was found to suppress ER, TGF-β, and NF-kB in-vitro gene expression strongly, and that was verified by qPCR and Western Blot gel electrophoresis techniques.

Keywords: HPLC-MS/MS, pomegranate, breast cancer, ovarian cancer, ER, TGF-β, NF-kB

Procedia PDF Downloads 71

Authors: Leto Leandra, Guaitini Caterina, Agosti Anna, Del Vecchio Lorenzo, Guarrasi Valeria, Cirlini Martina, Chiancone Benedetta

Abstract:

To the best of our knowledge, this study represents the first attempt to investigate the in vitro germination response of Rhus coriaria L., and its sprout chemical characterization. Rhus coriaria L., a species belonging to the Anacardiaceae family, is commonly called "sumac" and is cultivated, in different countries of the Mediterranean and the Middle East regions, to produce a spice with a sour taste, obtained from its dried and ground fruits. Moreover, since ancient times, many beneficial properties have been attributed to this plant that has been used, in the traditional medicine of several Asian countries, against various diseases, including liver and intestinal pathologies, ulcers and various inflammatory states. In the recent past, sumac was cultivated in the Southern regions of Italy to treat leather, but its cultivation was abandoned, and currently, sumac plants grow spontaneously in marginal areas. Recently, in Italy, the interest in this species has been growing again, thanks to its numerous properties; thus, it becomes imperative to deepen the knowledge of this plant. In this study, in order to set up an efficient in vitro seed germination protocol, sumac seeds collected from spontaneous plants grown in Sicily, an island in the South of Italy, were, firstly, subjected to different treatments, scarification (mechanical, physical and chemical), cold stratification and imbibition, to break their physical and physiological dormancy, then, treated seeds were in vitro cultured on media with different gibberellic acid (GA3) concentrations. Results showed that, without any treatment, only 5% of in vitro sown seeds germinated, while the germination percentage increased up to 19% after the mechanical scarification. A further significative improvement of germination percentages was recorded after the physical scarification, with (40.5%) or without (36.5%) 8 weeks of cold stratification, especially when seeds were sown on gibberellin enriched cultured media. Vitro-derived sumac sprouts, at different developmental stages, were chemically characterized, in terms of polyphenol and tannin content, as well as for their antioxidant activity, to evaluate this matrix as a potential novel food or as a source of bioactive compounds. Results evidenced how more developed sumac sprouts and, above all, their leaves are a wealthy source of polyphenols (78.4 GAE/g SS) and tannins (21.9 mg GAE/g SS), with marked antioxidant activity. The outcomes of this study will be of support the nursery sector and sumac growers in obtaining a higher number of plants in a shorter time; moreover, the sprout chemical characterization will contribute to the process of considering this matrix as a new source of bioactive compounds and tannins to be used in food and non-food sectors.

Keywords: bioactive compounds, germination pre-treatments, rhus coriaria l., tissue culture

Procedia PDF Downloads 59

Authors: Leto Leandra, Guaitini Caterina, Agosti Anna, Del Vecchio Lorenzo, Guarrasi Valeria, Cirlini Martina, Chiancone Benedetta

Abstract:

To the best of our knowledge, this study represents the first attempt to investigate the in vitro germination response of Rhus coriaria L. and its sprout chemical characterization. Rhus coriaria L., a species belonging to the Anacardiaceae family, is commonly called "sumac” and is cultivated, in different countries of the Mediterranean and the Middle East regions, to produce a spice with a sour taste, obtained from its dried and ground fruits. Moreover, since ancient times, many beneficial properties have been attributed to this plant that has been used, in the traditional medicine of several Asian countries, against various diseases, including liver and intestinal pathologies, ulcers, and various inflammatory states. In the recent past, sumac was cultivated in the Southern regions of Italy to treat leather, but its cultivation was abandoned, and currently, sumac plants grow spontaneously in marginal areas. Recently, in Italy, the interest in this species has been growing again, thanks to its numerous properties; thus, it becomes imperative to deepen the knowledge of this plant. In this study, in order to set up an efficient in vitro seed germination protocol, sumac seeds collected from spontaneous plants grown in Sicily, an island in the South of Italy, were, firstly, subjected to different treatments, scarification (mechanical, physical and chemical), cold stratification and imbibition, to break their physical and physiological dormancy, then, treated seeds were in vitro cultured on media with different gibberellic acid (GA3) concentrations. Results showed that, without any treatment, only 5% of in vitro sown seeds germinated, while the germination percentage increased up to 19% after the mechanical scarification. A further significative improvement of germination percentages was recorded after the physical scarification, with (40.5%) or without (36.5%) 8 weeks of cold stratification, especially when seeds were sown on gibberellin enriched cultured media. Vitro-derived sumac sprouts, at different developmental stages, were chemically characterized, in terms of polyphenol and tannin content, as well as for their antioxidant activity, to evaluate this matrix as a potential novel food or as a source of bioactive compounds. Results evidenced how more developed sumac sprouts and, above all, their leaves are a wealthy source of polyphenols (78.4 GAE/g SS) and tannins (21.9 mg GAE/g SS), with marked antioxidant activity. The outcomes of this study will be of support the nursery sector and sumac growers in obtaining a higher number of plants in a shorter time; moreover, the sprout chemical characterization will contribute to the process of considering this matrix as a new source of bioactive compounds and tannins to be used in food and non-food sectors.

Keywords: bioactive compounds, germination pre-treatments, rhus coriaria l., tissue culture

Procedia PDF Downloads 45

Authors: H. Pegram, R. Stevens, L. De Girolamo

Abstract:

Aligned electrospun nanofibres act as effective neuronal and glial cell scaffolds that can be layered to contain multiple sheets harboring different cell populations. This allows personalised biofunctional prostheses to be manufactured with both acellular and cellularised layers for the treatment of spinal cord injury. Additionally, the manufacturing route may be configured to produce in-vitro 3D cell based model of spinal cord injury to aid drug development and enhance prosthesis performance. The goal of this investigation was to optimise the multi-layer scaffold design parameters for prosthesis manufacture, to enable the development of multi-layer patient specific implant therapies. The work has also focused on the fabricating aligned nanofibre scaffolds that promote in-vitro neuronal and glial cell population growth, cell-to-cell interaction and long-term survival following trauma to mimic an in-vivo spinal cord lesion. The approach has established reproducible lesions and has identified markers of trauma and regeneration marked by effective neuronal migration across the lesion with glial support. The investigation has advanced the development of an in-vitro model of traumatic spinal cord injury and has identified a route to manufacture prostheses which target the repair spinal cord injury. Evidence collated to investigate the multi-layer concept suggests that physical cues provided by nanofibres provide both a natural extra-cellular matrix (ECM) like environment and controls cell proliferation and migration. Specifically, aligned nanofibre layers act as a guidance system for migrating and elongating neurons. On a larger scale, material type in multi-layer systems also has an influence in inter-layer migration as cell types favour different material types. Results have shown that layering nanofibre membranes create a multi-level scaffold system which can enhance or prohibit cell migration between layers. It is hypothesised that modifying nanofibre layer material permits control over neuronal/glial cell migration. Using this concept, layering of neuronal and glial cells has become possible, in the context of tissue engineering and also modelling in-vitro induced lesions.

Keywords: electrospinning, layering, lesion, modeling, nanofibre

Procedia PDF Downloads 96

Authors: Iram Siddique

Abstract:

Synthetic seed technology is an alternative to traditional micropropagation for production and delivery of cloned plantlets. Synthetic seeds were produced by encapsulating nodal segments of C. angustifolia in calcium alginate gel. 3% (w/v) sodium alginate and 100 mM CaCl2. 2H2O were found most suitable for encapsulation of nodal segments. Synthetic seeds cultured on half strength Murashige and Skoog (MS) medium supplemented with thidiazuron (5.0 µM) + indole -3- acetic acid (1.0 µM) produced maximum number of shoots (10.9 ± 0.78) after 8 weeks of culture exhibiting (78%) in vitro conversion response. Encapsulated nodal segments demonstrated successful regeneration after different period (1-6 weeks) of cold storage at 4 °C. The synthetic seeds stored at 4 °C for a period of 4 weeks resulted in maximum conversion frequency (93%) after 8 weeks when placed back to regeneration medium. The isolated shoots when cultured on half strength MS medium supplemented with 1.0 µM indole -3- butyric acid (IBA), produced healthy roots and plantlets with well developed shoot and roots were successfully hardened off in plastic pots containing sterile soilrite inside the growth chamber and gradually transferred to greenhouse where they grew well with 85% survival rate. Changes in the content of photosynthetic pigments, net photosynthetic rate (PN), superoxide dismutase (SOD) and catalase (CAT) activity in C. angustifolia indicated the adaptation of micropropagated plants to ex vitro conditions.

Keywords: biochemical studies, nodal segments, rooting, synthetic seeds, thidiazuron

Procedia PDF Downloads 332

Authors: Paresh M. Patel, Amit A. Patel, R. H. Parikh

Abstract:

The present investigation was undertaken to fabricate modified transport fluconazole that belongs to BCS class II and have a poor applicability on topical infection. So to improve topical application, transdermal spray could play a vital role by using ethyl cellulose and Eudragit® S100 as film-forming polymers. Concentration of Eudragit® S100, ethyl cellulose and permeation enhancer (camphor and menthol) were selected as independent variables, whereas drying time, viscosity and in-vitro drug release were selected as dependent variables in factorial design. The viscosity, drying time and in-vitro drug release of the optimize batch B15 was 40.1 cps, 47 sec. and 90.79% respectively. The film of optimized batch was flexible and dermal-adhesive.

Keywords: Eudragit, ethyl cellulose, fluconazole, transdermal spray

Procedia PDF Downloads 426

Authors: Muhammad Awais Afzal, Lorena Tuchscherr De Hauschopp, Christian Hübner

Abstract:

Introduction: Autophagy is an evolutionarily conserved lysosome-dependent degradation pathway, which can be induced by extrinsic and intrinsic stressors in living systems to adapt to fluctuating environmental conditions. In the context of inflammatory stress, autophagy contributes to the elimination of invading pathogens, the regulation of innate and adaptive immune mechanisms, and regulation of inflammasome activity as well as tissue damage repair. Lysosomes can be recycled from autolysosomes by the process of autophagic lysosome reformation (ALR), which depends on the presence of several proteins including Spatacsin. Thus ALR contributes to the replenishment of lysosomes that are available for fusion with autophagosomes in situations of increased autophagic turnover, e.g., during bacterial infections, inflammatory stress or sepsis. Objectives: We aimed to assess whether ALR plays a role for cell survival in an in-vitro bacterial infection model. Methods: Mouse embryonic fibroblasts (MEFs) were isolated from wild-type mice and Spatacsin (Spg11-/-) knockout mice. Wild-type MEFs and Spg11-/- MEFs were infected with Staphylococcus aureus (multiplication of infection (MOI) used was 10). After 8 and 16 hours of infection, cell viability was assessed on BD flow cytometer through propidium iodide intake. Bacterial intake by cells was also calculated by plating cell lysates on blood agar plates. Results: in-vitro infection of MEFs with Staphylococcus aureus showed a marked decrease of cell viability in ALR deficient Spatacsin knockout (Spg11-/-) MEFs after 16 hours of infection as compared to wild-type MEFs (n=3 independent experiments; p < 0.0001) although no difference was observed for bacterial intake by both genotypes. Conclusion: Suggesting that ALR is important for the defense of invading pathogens e.g. S. aureus, we observed a marked increase of cell death in an in-vitro infection model in cells with compromised ALR.

Keywords: autophagy, autophagic lysosome reformation, bacterial infections, Staphylococcus aureus

Procedia PDF Downloads 106

Authors: Hossein Mostafavi

Abstract:

A diversity of biological activities and pharmaceutical uses have been attributed to gallic acid derivatives such as antibacterial, anticancer, anti inflammatory. A series of gallic acid derivatives were synthesized, and their structure was confirmed by FT-IR, HNMR, CNMR, elemental analysis. In vitro biological activity of compounds was determined against Proteus vulgaris ATCC 7829, Escherichia coli ATCC 25922, as (Gram-negative) bacteria and bacillus cereus ATCC 11778, Staphylococus aureus ATCC 6538 as (Gram-positive) bacteria. Antibacterial susceptibility tests were done by use of the paper disc diffusion method on Mueller Hinton agar (Merck). Chloramiphenicol, Penicilline, Streptomycin and Tetracycline were standard reference antibiotics. The zone of inhibition against bacteria was measured after 24 hours at 37 °C. Compounds 3, 4, 5 were the main antibacterial compounds against Gram-negative bacteria but not Gram-positive.

Keywords: gallic acid derivatives, antibacterial, antibiotics, inhibition

Procedia PDF Downloads 105

Authors: Umme Hani, Mohammed Rahmatulla, Mohammed Ghazwani, Ali Alqahtani, Yahya Alhamhoom

Abstract:

Background and introduction: Neratinib is a potent anticancer drug used for the treatment of breast cancer. It is poorly soluble at higher pH, which tends to minimize the therapeutic effects in the lower gastrointestinal tract (GIT) leading to poor bioavailability. An attempt has been made to prepare and develop a gastro-retentive system of Neratinib to improve the drug bioavailability in the GIT by enhancing the gastric retention time. Materials and methods: In the present study a three-factor at two-level (23) factorial design based optimization was used to inspect the effects of three independent variables (factors) such as sodium alginate (A), sodium bicarbonate (B) and sodium citrate (C) on the dependent variables like in vitro gelation, in vitro floating, water uptake and percentage drug release. Results: All the formulations showed pH in the range 6.7 ±0.25 to 7.4 ±0.24, percentage drug content was observed to be 96.3±0.27 to 99.5 ±0.28%, in vitro gelation observed as gelation immediate remains for an extended period. Percentage of water uptake was in the range between 9.01±0.15 to 31.01±0.25%, floating lag time was estimated form 7±0.39 to 57±0.36 sec. F4 and F5 showed floating even after 12hrs. All formulations showed a release of around 90% drug release within 12hr. It was observed that the selected independent variables affect the dependent variables. Conclusion: The developed system may be a promising and alternative approach to augment gastric retention of drugs and enhances the therapeutic efficacy of the drug.

Keywords: neratinib, 2³ factorial design, sodium alginate, floating, in situ gelling system

Procedia PDF Downloads 115

Authors: Carolina S. Monteiro, Eugénia Pinto, Miguel A. Faria, Sara C. Cunha

Abstract:

About half of the world's population eats rice daily, making it the primary food source for billions of people. Besides its nutrition potential, rice can be a significant route of exposure to many contaminants. Mycotoxins are an example of such contaminants that can be present in rice. Among them, ochratoxin (OTA), citrinin (CIT), and zearalenone (ZEN) are frequently reported in rice. During digestion, only a fraction of mycotoxins from food can be absorbed (bioaccessible fraction), influencing their ability to cause toxic effects. Insufficient knowledge of the bioavailability of mycotoxins, alone and in combination, may hinder an accurate risk assessment of contaminants ingested by humans. In this context, two different rice (Oryza sativa) varieties, Carolino white and Carolino brown, both with and without turmeric, were boiled and individually spiked with OTA, CIT, and ZEN plus with its combination. Subsequently, samples were submitted to the INFOGEST harmonized in vitro digestion protocol to evaluate the bioaccessibility of mycotoxins. Afterward, the in vitro intestinal transport of the mycotoxins, both alone and in combination, was evaluated in digests of Carolino white rice with and without turmeric. Assays were performed with a monolayers of of Caco-2 and HT-29 cells. Bioaccessibility of OTA and ZEN, alone and in combination, were similar in Carolino white and brown rice with or without turmeric. For CIT, when Carolino white rice was used, the bioaccessibility was higher alone than in combination (62.00% vs. 25.00%, without turmeric; 87.56% vs. 53.87%, with turmeric); however, with Carolino brown rice was the opposite (66.38% vs. 75.20%, without turmeric; 43.89% vs. 59.44%, with turmeric). All the mycotoxins, isolated, reached the higher bioaccessibility in the Carolino white rice with turmeric (CIT: 87.56%; OTA: 59.24%; ZEN: 58.05%). When mycotoxins are co-present, the higher bioaccessibility of each one varies with the type of rice. In general, when turmeric is present, bioaccessibility increases, except for CIT, using Carolino brown rice. Concerning the intestinal absorption in vitro, after 3 hours of transport, all mycotoxins were detected in the basolateral compartment being thus transported through the cells monolayer. ZEN presented the highest fraction absorbed isolated and combined, followed by CIT and OTA. These findings highlight that the presence of other components in the complex dietary matrix, like turmeric, and the co-presence of mycotoxins can affect its final bioavailability with obvious implications for health risk. This work provides new insights to qualitatively and quantitatively describe mycotoxin in rice fate during human digestion and intestinal absorption and further contribute to better risk assessment.

Keywords: bioaccessibility, digestion, intestinal absorption, mycotoxins

Procedia PDF Downloads 26

Authors: Mohamed E. Khalifa, Adil A. Gobouri

Abstract:

Potential synthesis of a series of 3-amino-4-arylazothiophene derivatives from reaction of 2-cyano-2-phenylthiocarbamoyl acetamide and the appropriate α-halogenated reagents, followed by coupling with different aryl diazonium salts (Japp-Klingemann reaction), and another series of 5-arylazo-thiazol-2-ylcarbamoyl-thiophene derivatives from base-catalyzed intramolecular condensation of 5-arylazo-2-(N-chloroacetyl)amino-thiazole with selected B-keto compounds (Thorpe-Ziegler reaction) was performed. The biological activity of the two series was studied in vitro. Their versatility for pharmaceutical purposes was reported, where they displayed remarkable activities against selected pathogenic microorganisms; Bacillus subtilize, Staphylococcus aureus (Gram positive bacteria), Escherichia coli, Pseudomonas aeruginosa (Gram negative bacteria) and Aspergillus flavus, Candida albicans (fungi) with various degrees related to their chemical structures.

Keywords: thiophene, 2-aminothiazole, compounds, antioxidant, antitumor, antimicrobial

Procedia PDF Downloads 313

Authors: Wassim Guermazi, Saoussan Boukhris, Neila Annabi Trabelsi, Tarek Rebai, Alya Sellami-Kamoun, Habib Ayadi

Abstract:

This work investigates the protective effects of the microalga Halamphora sp. extract (H. Ext) as a natural product on lead-intoxicated liver and kidney human cells in vitro and in vivo on rats wistar. HepG2 cells line derived from human hepatocellular carcinoma and HEK293 cells line derived from human embryonic kidney were used for the in vitro study. The analysis of the fatty acids methyl esters of the extract was performed by a GC/MS. Four groups of rats, each of which was composed of six animals, were used for the in vivo experiment. The pretreatment of HepG2 and HEK293 cells line with the extract (100 µg mL-1) significantly (p < 0.05) protected against cytotoxicity induced by lead exposure. In vivo, the biochemical parameters in serum, namely malondialdehyde level (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activities, were measured in supernatants of organ homogenates. H. Ext was found to be rich in fatty acids, essentially palmitic and palmitoleic accounting respectively 29.46% and 42.07% of total fatty acids. Both in vitro and in vivo, the co-treatment with H. Ext allowed the protection of the liver and kidney cells structure, as well as the significant preservation of normal antioxidant and biochemical parameters in rats. Halamphora extract rich in fatty acids has been proven to be effective in protection against Pb-induced toxicity.

Keywords: microalga extract, human cells line, fatty acid, lead exposure, oxidative stress, rats

Procedia PDF Downloads 53

Authors: Dinakaran Venkatachalam, Paul Chambers, Kavitha Kongara, Preet Singh

Abstract:

The objective of this study is to formulate different topical preparations containing articaine and to investigate their permeation through goat skin. Initially, articaine and its hydrochloride salt were compared for in vitro permeation using Franz cell model. Goat skin samples were collected after euthanizing male goat kids purchased from the dairy goat farmers. Subcutaneous fat was removed and the skin was mounted on the donor chamber (orifice area 1.00 cm²) and drugs were applied onto the epidermis. Phosphate buffer saline (pH 7.4) was used to maintain sink condition in the receptor chamber (8 ml) of the Franz cell. Samples (0.4 ml) were collected at various intervals over 24 hours after each sampling equal volume of PBS was replaced in the receptor chamber. Articaine in the collected samples were quantified using LC/MS. The results suggested that articaine free base permeates better than its hydrochloride salt through goat skin. This study results support the fact that local anesthetics in its base form are lipophilic and thus penetrates faster through cell membranes than their salts. Later, articaine free base was formulated either using ethanol and octyl salicylate or dimethyl sulfoxide (DMSO) as penetration enhancers and was compared for in vitro permeation. The transdermal flux of articaine in the formulation containing DMSO was approximately 3.8 times higher than that of the formulation containing ethanol and octyl salicylate. Further studies to evaluate the local anesthetic efficacy of the topical formulation containing articaine for dermal anesthesia in animals have been planned.

Keywords: articaine, dermal anesthesia, local anesthetic, transdermal

Procedia PDF Downloads 207

Authors: Emílie Kučerová, Tereza Veverková, Marina Morozovová, Eva Kudová, Jitka Viktorová

Abstract:

The blood-brain barrier (BBB) is a key element regulating the transport of substances between the blood and the central nervous system (CNS). The BBB protects the CNS from potentially harmful substances and maintains a suitable environment for nervous activity in the CNS, but at the same time, it represents a significant obstacle to the entry of drugs into the CNS. Pharmacoresistant epilepsy is a form of epilepsy that cannot be suppressed using two (or more) appropriately chosen antiepileptic drugs. In many cases, pharmacoresistant epilepsy is characterized by an increased concentration of efflux pumps on the luminal sides of the endothelial cells that form the BBB and an increased number of drug-metabolizing enzymes in the BBB cells, thereby preventing the effective transport of antiepileptic drugs into the CNS. Currently, a number of scientific groups are focusing on the preparation and improvement of BBB models in vitro in order to study cell interactions or transport mechanisms. However, in pathological conditions such as pharmacoresistant epilepsy, there are changes in BBB structure, and current BBB models are insufficient for related research. Our goal is to develop a suitable BBB model for pharmacoresistant epilepsy in vitro and use it to test the transfer of potential antiepileptic drugs. This model is created by co-culturing immortalized human cerebral microvascular endothelial cells, human vascular pericytes and immortalized human astrocytes. The BBB in vitro is cultivated in the form of a 2D transwell model and the integrity of the barrier is verified by measuring transendothelial electrical resistance (TEER). From the current results, a contact cell arrangement with the cultivation of endothelial cells on the upper side of the insert and the co-cultivation of astrocytes and pericytes on the lower side of the insert is selected as the most promising for BBB model cultivation. The pharmacoresistance of the BBB model is achieved by long-term cultivation of endothelial cells in an increasing concentration of selected antiepileptic drugs, which should lead to increased production of efflux pumps and drug-metabolizing enzymes. The pharmacoresistant BBB model in vitro will be further used for the screening of substances that could act both as antiepileptics and at the same time as inhibitors of efflux pumps in endothelial cells. This project was supported by the Technology Agency of the Czech Republic (TACR), Personalized Medicine: Translational research towards biomedical applications, No. TN02000109 and by the Academy of Sciences of the Czech Republic (AS CR) – grant RVO 61388963.

Keywords: antiepileptic drugs, blood-brain barrier, efflux transporters, pharmacoresistance

Procedia PDF Downloads 24

Authors: T. Scattolin, E. Cavarzerani, F. Visentin, F. Rizzolio

Abstract:

Organopalladium compounds have recently attracted attention for their high stability even under physiological conditions and, above all, for their remarkable in vitro cytotoxicity towards cisplatin-resistant cell lines. Among the organopalladium derivatives, those bearing at least one N-heterocyclic carbene ligand (NHC) and the Pd(II)-η³-allyl fragment have exhibited IC₅₀ values in the micro and sub-micromolar range towards several cancer cell lines in vitro and in some cases selectivity towards cancerous vs. non-tumorigenic cells. Herein, a selection of allyl and indenyl palladates were synthesized using a solvent-free method consisting of grinding the corresponding palladium precursors with different saturated and unsaturated azolium salts. All compounds have been fully characterized by NMR, XRD and elemental analyses. The intramolecular H, Cl interaction has been elucidated and quantified using the Voronoi Deformation Density scheme. Most of the complexes showed excellent cytotoxicity towards ovarian cancer cell lines, with I₅₀ values comparable to or even lower than cisplatin. Interestingly, the potent anticancer activity was also confirmed in a high-serous ovarian cancer (HGSOC) patient-derived tumoroid, with a clear superiority of this class of compounds over classical platinum-based agents. Finally, preliminary enzyme inhibition studies of the synthesized palladate complexes against the model TrxR show that the compounds have high activity comparable to or even higher than auranofin and classical Au(I) NHC complexes. Based on such promising data, further in vitro and in vivo experiments and in-depth mechanistic studies are ongoing in our laboratories.

Keywords: anticancer activity, palladium complexes, organoids, indenyl and allyl ligands

Procedia PDF Downloads 64

Authors: F. Belkhiri, A. Baghiani, S. Boumerfeg, N. Charef, S. Khennouf, L. Arrar

Abstract:

The present study was conducted to evaluate the in vitro antioxidant and antibacterial properties of methanolic extracts from roots of Tamus communis L. (TCRE), which is a plant used in traditional medicine in Algeria. The antioxidant potential of pattern was evaluated using tow complementary techniques, inhibition of free radical DPPH and the test of β-Carotene/linoleic acid. The antioxidant test indicates that non-polar fractions of TCRE (chloroform and ethyl acetate fractions) were more active than the polar fractions. Among these fractions, the chloroform extract appear in the DPPH test an IC50 of (18.89 µg/ml) comparable to that of BHT (18.6 µg/ml). This fraction was able to inhibiting the oxidation of β-Carotene with a percentage of inhibition (89.84 %). In antibacterial test, non-polar fractions showed antibacterial activity very important compared with the polar fractions. These fractions have inhibited the growth of four from nine bacterial strains, causing zones of inhibition from 08 to 23 mm of diameter.

Keywords: antioxidant activity, antibacterial activity, Tamus communis L., polar fractions

Procedia PDF Downloads 542

Authors: Djemai Zoughlache Soumia, Yahia Mouloud, Lekbir Adel, Meslem Meriem, Maouchi Madiha, Bahi Ahlem, Benbia Souhila

Abstract:

Natural extracts is known for their contents of biologically active molecules. In this context, we attempted to evaluate the antioxidant activity of the methanolic extract prepared from the bark of the roots of Zizyphus lotus. The quantitative analysis based on the dosage, phenolic compounds, flavonoids and tannins provided following values: 0.39 ± 0.007 ug EAG/mg of extract for phenolic compounds, 0.05 ± 0.02ug EQ/mg extract for flavonoids and 0.0025 ± 7.071 E-4 ECT ug/mg extract for tannins. The study of the antioxidant activity by the DPPH test in vitro showed a powerful antiradical power with an IC50 = 8,8 ug/ml. For the DPPH test in vivo we used two rats lots, one lot with a dose of 200 mg/kg of the methanol extract and a control lot. We found a significant difference in antiradical activity with p < 0.05.

Keywords: Zizyphus lotus, antioxidant activity, DPPH, phenolic compounds, flavonoids, tannins

Procedia PDF Downloads 473

Authors: P. Karpagam, Babu Joseph, P. Ashok Kumar

Abstract:

The incidence of Candida infections is increasing worldwide. The serious nature of these infections is compounded by increasing levels of drug resistance. Pure cultures of the Candida sp. were obtained from clinical isolates and fresh garlic extracts were obtained by extraction techniques. The antifungal activity of garlic extract was investigated in an in vitro system. The extract (100%, 75% and 50%) showed significant antifungal activity against Candida, whereas, low concentration (25%) of the extract showed less antifungal activity against the test organism. Antifungal activities of honey and lemon juice were tested against the Candida; however, the growth was not inhibited by these extracts. On the other hand honey and lemon when combined with garlic exhibited a good antifungal activity. The study thus confirms the antifungal properties of garlic extract along with additives like honey and lemon have significant antifungal activity against isolates of Candida species.

Keywords: Candida, garlic extract, lemon, synergitic antifungal activity

Procedia PDF Downloads 212

Authors: Dupe Olufunke Ogunbosoye, Thaofik Badmos Mustapha, Lanre Shaffihy Adeaga, R. O. Imam

Abstract:

Efforts have been made to reduce ruminants' methane emissions while improving animal productivity. Hence, an experiment was conducted to investigate the in vitro fermentation pattern, methane production, and performance of West African dwarf (WAD) rams-fed diets at graded levels of ginger. Sixteen (16) rams were randomly allocated into four dietary treatments with four animals per treatment in a completely randomized design for 84 days. Ginger powder was added at 0.00%, 0.25%, 0.50% and 0.75% as T1, T2, T3 and T4 respectively. The results indicated that at the 24-hour diet incubation, gas production, methane, metabolizable energy (ME), organic matter digestibility (OMD), and short-chain fatty acids (SCFA) concentrations decreased with the increasing level of ginger. Conversely, the sheep-fed T4 recorded the highest daily weight gain (47.61g/day), while the least daily weight gain (17.86g/day) was recorded in ram-fed T1. The daily weight gain of the rams fed T3 and T4 was similar but significantly different from the daily weight gain in T1 (17.86g/day) and T2 (29.76g/day). Daily feed intake was not significantly different across the treatments. T4 recorded the best response regarding feed conversion ratio (18.59) compared with other treatments. Based on the results obtained, rams fed T4 perform best in terms of growth and methane production. It is therefore concluded that the addition of ginger powder into the diet of sheep up to 0.75% enhances the growth rate of WAD sheep and reduces enteric methane production to create a smart nutrition system in ruminant animal production.

Keywords: enteric methane, growth, in vitro, sheep, nutrition system

Procedia PDF Downloads 48

Authors: Arash Khorasani Esmaeili, Rosna Mat Taha, Sadegh Mohajer

Abstract:

Seeds of Trifolium pratense (Red clover) were exposed in vitro for 6 weeks to six levels of lead (Pb) concentrations (0, 50, 100, 150, 200, 250 µM) to analyze the effects on growth, total chlorophyll and total protein contents of grown plants against the lead accumulation. The growth of plants was negatively affected by various levels of lead treatment. The fresh and dry weights, as well as lengths of shoots and roots of grown plants under various lead treatments, were found significantly lower in comparison with the control plants. Total chlorophyll and total soluble protein contents of grown plants under lower concentrations of lead treatment did not show significant differences when compared with the control plants, although they were affected significantly in higher levels of lead accumulation (150-250 µM).

Keywords: trifolium pratense, lead accumulation, chlorophyll content, protein content

Procedia PDF Downloads 406

Authors: Nacera Baali, Zahia Belloum, Souad Ameddah, Fadila Benayache, Samir Benayache, Chantal Wrutniak-Cabello

Abstract:

Allurement of herbs as health beneficial foods and as a source material for the development of new drugs, has led to greater furtherance in the study of herbal medicines during recent years. In the present study, in vitro antioxidant, free radical scavenging capacity, and hepatoprotective activity of butanolic extract from Genista quadriflora Munby (G.quadriflora) were evaluated using established in vitro models such as DPPH radical and hydrogen peroxide radical scavenging activities and antilipidperoxidation ability. Interestingly, the extract showed considerable in vitro antioxidant and free radical scavenging activities in a dose-dependent manner when compared to the standard antioxidant which verified the presence of antioxidant compound in extract tested. The hepatoprotective potential of G.quadriflora extract was also evaluated in male Wistar rats against paracetamol (APAP) induced liver damage. Therapy of G.quadriflora showed the liver protective effect on biochemical and histopathological alterations. Moreover, histological studies also supported the biochemical finding, that is, the maximum improvement in the histoarchitecture of the liver. Results revealed that G.quadriflora extract could protect the liver against APAP-induced oxidative damage by possibly increasing the antioxidant protection mechanism in rats. These findings are of great importance in view of the availability of the plant and its observed possible diverse applications in medicine and nutrition.

Keywords: genista quadriflora munby, antioxidant, liver, paracetamol, oxidative stress

Procedia PDF Downloads 436