Search results for: corneal epithelial cells

Authors: Chia-Jung Li, Kuan-Hao Tsui

Abstract:

As women age, the quality of their oocytes deteriorates irreversibly, leading to reduced fertility. To better understand the role of Ferroptosis-related genes in ovarian aging, we employed a multi-omics analysis approach, including spatial transcriptomics, single-cell RNA sequencing, human ovarian pathology, and clinical biopsies. Our study identified excess lipid peroxide accumulation in aging germ cells, metal ion accumulation via oxidative reduction, and the interaction between ferroptosis and cellular energy metabolism. We used multi-histological prediction of ferroptosis key genes to evaluate 75 patients with ovarian aging insufficiency and then analyzed changes in hub genes after supplementing with DHEA, Ubiquinol CoQ10, and Cleo-20 T3 for two months. Our results demonstrated a significant increase in TFRC, GPX4, NCOA4, and SLC3A2, which were consistent with our multi-component prediction. We theorized that these supplements increase the mitochondrial tricarboxylic acid cycle (TCA) or electron transport chain (ETC), thereby increasing antioxidant enzyme GPX4 levels and reducing lipid peroxide accumulation and ferroptosis. Overall, our findings suggest that supplementation intervention significantly improves IVF outcomes in senescent cells by enhancing metal ion and energy metabolism and enhancing oocyte quality in aging women.

Keywords: multi-omics, nutrients, ferroptosis, ovarian aging

Procedia PDF Downloads 77

Authors: Roman Matejka, Vaclav Prochazka, Tibor Izak, Jana Stepanovska, Martina Travnickova, Alexander Kromka

Abstract:

Cell-based impedance spectroscopy is suitable method for electrical monitoring of cell activity especially on substrates that cannot be easily inspected by optical microscope (without fluorescent markers) like decellularized tissues, nano-fibrous scaffold etc. Special sensor for this measurement was developed. This sensor consists of corning glass substrate with gold interdigitated electrodes covered with diamond layer. This diamond layer provides biocompatible non-conductive surface for cells. Also, a special PPFC flow cultivation chamber was developed. This chamber is able to fix sensor in place. The spring contacts are connecting sensor pads with external measuring device. Construction allows real-time live cell imaging. Combining with perfusion system allows medium circulation and generating shear stress stimulation. Experimental evaluation consist of several setups, including pure sensor without any coating and also collagen and fibrin coating was done. The Adipose derived stem cells (ASC) and Human umbilical vein endothelial cells (HUVEC) were seeded onto sensor in cultivation chamber. Then the chamber was installed into microscope system for live-cell imaging. The impedance measurement was utilized by vector impedance analyzer. The measured range was from 10 Hz to 40 kHz. These impedance measurements were correlated with live-cell microscopic imaging and immunofluorescent staining. Data analysis of measured signals showed response to cell adhesion of substrates, their proliferation and also change after shear stress stimulation which are important parameters during cultivation. Further experiments plan to use decellularized tissue as scaffold fixed on sensor. This kind of impedance sensor can provide feedback about cell culture conditions on opaque surfaces and scaffolds that can be used in tissue engineering in development artificial prostheses. This work was supported by the Ministry of Health, grants No. 15-29153A and 15-33018A.

Keywords: bio-impedance measuring, bioreactor, cell cultivation, diamond layer, gold interdigitated electrodes, tissue engineering

Procedia PDF Downloads 285

Authors: Rupinder Kaur, Parmjit S. Panesar, Ram S. Singh

Abstract:

Whey is the lactose rich by-product of the dairy industry, having good amount of nutrient reservoir. Most abundant nutrients are lactose, soluble proteins, lipids and mineral salts. Disposing of whey by most of milk plants which do not have proper pre-treatment system is the major issue. As a result of which, there can be significant loss of potential food and energy source. Thus, whey has been explored as the substrate for the synthesis of different value added products such as enzymes. β-galactosidase is one of the important enzymes and has become the major focus of research due to its ability to catalyze both hydrolytic as well as transgalactosylation reaction simultaneously. The enzyme is widely used in dairy industry as it catalyzes the transformation of lactose to glucose and galactose, making it suitable for the lactose intolerant people. The enzyme is intracellular in both bacteria and yeast, whereas for molds, it has an extracellular location. The present work was carried to utilize the whey for the production of β-galactosidase enzyme using both yeast and fungal cultures. The yeast isolate Kluyveromyces marxianus WIG2 and various fungal strains have been used in the present study. Different disruption techniques have also been investigated for the extraction of the enzyme produced intracellularly from yeast cells. Among the different methods tested for the disruption of yeast cells, SDS-chloroform showed the maximum β-galactosidase activity. In case of the tested fungal cultures, Aureobasidium pullulans NCIM 1050, was observed to be the maximum extracellular enzyme producer.

Keywords: β-galactosidase, fungus, yeast, whey

Procedia PDF Downloads 308

Authors: Yuliya Markava

Abstract:

The laser in situ keratomileusis (LASIK) is widely common surgical procedure, which has become an alternative for patients who are not satisfied with the performance of other correction methods. A high level of patient satisfaction functional outcomes after refractive surgery confirms the high reliability and safety of LASIK and provides a significant improvement in the quality of life and social adaptation. Purpose: To perform clinical analysis of the results of correction made to the excimer laser system SCHWIND AMARIS 500E in patients with different types of refractive anomalies. Materials and Methods: This was a retrospective analysis of 1581 operations (812 patients): 413 males (50.86%) and 399 females (49.14%) at the age from 18 to 47 years with different types of ametropia. All operations were performed on excimer laser SCHWIND AMARIS 500E in the LASIK procedure. Formation of the corneal flap was made by mechanical microkeratome SCHWIND. Results: Analyzing the structure of refractive anomalies: The largest number of interventions accounted for myopia: 1505 eyes (95.2%), of which about a low myopia: 706 eyes (44.7%), moderate myopia: 562 eyes (35.5 %), high myopia: eyes 217 (13.7%) and supermyopia: 20 eyes (1.3%). Hyperopia was 0.7% (11 eyes), mixed astigmatism: 4.1% (65 eyes). The efficiency was 80% (in patients with supermyopia) to 91.6% and 95.4% (in groups with myopia low and moderate, respectively). Uncorrected visual acuity average values before and after laser operation was in groups: a low myopia 0.18 (up 0.05 to 0.31) and 0.80 (up 0.60 to 1.0); moderate myopia 0.08 (up 0.03 to 0.13) and 0.87 ( up 0.74 to 1.0); high myopia 0.05 (up 0.02 to 0.08) and 0.83 (up 0.66 to 1.0); supermyopia 0.03 (up 0.02 to 0.04) and 0.59 ( up 0.34 to 0.84); hyperopia 0.27 (up 0.16 to 0.38) and 0.57 (up 0.27 to 0.87); mixed astigmatism of 0.35 (up 0.19 to 0.51) and 0.69 (up 0.44 to 0.94). In all cases, after LASIK indicators uncorrected visual acuity significantly increased. Reoperation was 4.43%. Significance: Clinical results of refractive surgery at the excimer laser system SCHWIND AMARIS 500E in different ametropia correction is characterized by high efficiency.

Keywords: effectiveness of laser correction, LASIK, refractive anomalies, surgical treatment

Procedia PDF Downloads 236

Authors: M. I. Tyletc, O. I. Podgornya, T. G. Shaposhnikova, S. V. Shabelnikov, A. G. Mittenberg, M. A. Daugavet

Abstract:

Ascidiacea is the most numerous class of the Tunicata subtype. These chordates' distinctive feature of the anatomical structure is a tunic consisting of cellulose fibrils, protein molecules, and single cells. The mechanisms of the tunic formation are not known in detail; tunic formation could be used as the model system for studying the interaction of cells with the extracellular matrix. Our model species is the ascidian Styela rustica, which is prevalent in benthic communities of the White Sea. As previously shown, the tunic formation involves morula blood cells, which contain the major 48 kDa protein p48. P48 participation in the tunic formation was proved using antibodies against the protein. The nature of the protein and its function remains unknown. The current research aims to determine the amino acid sequence of p48, as well as to clarify its role in the tunic formation. The peptides that make up the p48 amino acid sequence were determined by mass spectrometry. A search for peptides in protein sequence databases identified sequences homologous to p48 in Styela clava, Styela plicata, and Styela canopus. Based on sequence alignment, their level of similarity was determined as 81-87%. The correspondent sequence of ascidian Styela canopus was used for further analysis. The Styela rustica p48 sequence begins with a signal peptide, which could indicate that the protein is secretory. This is consistent with experimentally obtained data: the contents of morula cells secreted in the tunic matrix. The isoelectric point of p48 is 9.77, which is consistent with the experimental results of acid electrophoresis of morula cell proteins. However, the molecular weight of the amino acid sequence of ascidian Styela canopus is 103 kDa, so p48 of Styela rustica is a shorter homolog. The search for conservative functional domains revealed the presence of two Ca-binding EGF-like domains, thrombospondin (TSP1) and tyrosinase domains. The p48 peptides determined by mass spectrometry fall into the region of the sequence corresponding to the last two domains and have amino acid substitutions as compared to Styela canopus homolog. The tyrosinase domain (pfam00264) is known to be part of the phenoloxidase enzyme, which participates in melanization processes and the immune response. The thrombospondin domain (smart00209) interacts with a wide range of proteins, and is involved in several biological processes, including coagulation, cell adhesion, modulation of intercellular and cell-matrix interactions, angiogenesis, wound healing and tissue remodeling. It can be assumed that the tyrosinase domain in p48 plays the role of the phenoloxidase enzyme, and TSP1 provides a link between the extracellular matrix and cell surface receptors, and may also be responsible for the repair of the tunic. The results obtained are consistent with experimental data on p48. The domain organization of protein suggests that p48 is an enzyme involved in the tunic tunning and is an important regulator of the organization of the extracellular matrix.

Keywords: ascidian, p48, thrombospondin, tyrosinase, tunic, tunning

Procedia PDF Downloads 94

Authors: Taha Kadir Yesin, Hanyu Liu, Zhangfan Ding, Amit Singh, Qi Tian, Yuheng Zhang, Biswajyoti Borah, Junyu Chen, Anjali P. Kusumbe

Abstract:

The skeletal structure and bone marrow endothelium collectively form a critical functional unit essential for bone development, health, and aging. At the core of osteogenesis and bone formation lies the dynamic process of angiogenesis. In this study, we reveal a potent endogenous anabolic NCAM1-14.3.3. ζδ-derived- Peptide interaction, which stimulates bone angiogenesis and osteogenesis during homeostasis, aging, and age-related bone diseases. Employing high-resolution imaging and inducible cell-specific mouse genetics, our results elucidate the pivotal role of the NCAM1-14.3.3.ζδ-derived-Peptide interaction in driving the expansion of Clec14a+ angiogenic endothelial cells. Notably, Clec14a+ endothelial cells express key osteogenic factors. The NCAM1-14.3.3.ζδ-derived-Peptide interaction in osteoblasts drives osteoblast differentiation, ultimately contributing to the genesis of bone. Moreover, the NCAM1-14.3.3.ζδ-derived-Peptide interaction leads to a reduction in bone resorption. In age-associated vascular and bone loss diseases, stimulating the NCAM1-14.3.3.ζδ-derived-Peptide interaction not only promotes angiogenesis but also reverses bone loss. Consequently, harnessing the endogenous anabolic potential of the NCAM1-14.3.3.ζδ-derived-Peptide interaction emerges as a promising therapeutic modality for managing age-related bone diseases.

Keywords: endothelial cell, NCAM1, Clec14a, 14.3.3.ζδ

Procedia PDF Downloads 42

Authors: M. C. O. Ezeibe, F. I. O. Ezeibe

Abstract:

Reasons viral diseases (including AI, HIV/AIDS, and COVID-19), tumors (including Cancers and Prostrate enlargement), and antimicrobial-resistant infections (AMR) are difficult to cure are features of the pathogens which normal cells do not have or need (biomedical markers) have not been identified; medicines that can counter the markers have not been invented; strategies and mechanisms for their treatments have not been developed. When cells become abnormal, they acquire negative electrical charges, and viruses are either positively charged or negatively charged, while normal cells remain neutral (without electrical charges). So, opposite charges' electrostatic attraction is a treatment mechanism for viral diseases and tumors. Medicines that have positive electrical charges would mop abnormal (infected and tumor) cells and DNA viruses (negatively charged), while negatively charged medicines would mop RNA viruses (positively charged). Molecules of Aluminum-magnesium silicate [AMS: Al₂Mg₃ (SiO₄)₃], an approved medicine and pharmaceutical stabilizing agent, consist of nanoparticles which have both positive electrically charged ends and negative electrically charged ends. The very small size (0.96 nm) of the nanoparticles allows them to reach all cells in every organ. By stabilizing antimicrobials, AMS reduces the rate at which the body metabolizes them so that they remain at high concentrations for extended periods. When drugs remain at high concentrations for longer periods, their efficacies improve. Again, nanoparticles enhance the delivery of medicines to effect targets. Both remaining at high concentrations for longer periods and better delivery to effect targets improve efficacy and make lower doses achieve desired effects so that side effects of medicines are reduced to allow the immunity of patients to be enhanced. Silicates also enhance the immune responses of treated patients. Improving antimicrobial efficacies and enhancing patients` immunity terminate infections so that none remains that could develop resistance. Some countries do not have natural deposits of AMS, but they may have Aluminum silicate (AS: Al₄ (SiO₄)₃) and Magnesium silicate (MS: Mg₂SiO₄), which are also approved medicines. So, AS and MS were used to formulate an AMS-brand, named Medicinal synthetic AMS {Al₄ (SiO₄)₃ + 3Mg₂SiO₄ → 2Al₂Mg₃ (SiO₄)₃}. To overcome the challenge of AMS, AS, and MS being un-absorbable, Dextrose monohydrate is incorporated in MSAMS-formulations for the simple sugar to convey the electrically charged nanoparticles into blood circulation by the principle of active transport so that MSAMS-antimicrobial formulations function systemically. In vitro, MSAMS reduced (P≤0.05) titers of viruses, including Avian influenza virus and HIV. When used to treat virus-infected animals, it cured Newcastle disease and Infectious bursa disease of chickens, Parvovirus disease of dogs, and Peste des petits ruminants disease of sheep and goats. A number of HIV/AIDS patients treated with it have been reported to become HIV-negative (antibody and antigen). COVID-19 patients are also reported to recover and test virus negative when treated with MSAMS. PSA titers of prostate cancer/enlargement patients normalize (≤4) following treatment with MSAMS. MSAMS has also potentiated ampicillin trihydrate, sulfadimidin, cotrimoxazole, piparazine citrate and chloroquine phosphate to achieve ≥ 95 % infection-load reductions (AMR-prevention). At 75 % of doses of ampicillin, cotrimoxazole, and streptomycin, supporting MSAMS-formulations' treatments with antioxidants led to the termination of even already resistant infections.

Keywords: electrical charges, viruses, abnormal cells, aluminum-magnesium silicate

Procedia PDF Downloads 47

Authors: Thilivhali Emmanuel Tshikalange, Phiwokuhle Mamba

Abstract:

Ten medicinal plants used traditionally in the treatment of sexually transmitted diseases (STDs) and urinary tract infections (UTIs) were selected from an ethnobotanical database developed in Mpumalanga. The plants were investigated for their antimicrobial activity against five bacterial strains (Escherichia coli, Klebsiella oxytoca, Klebsiella pneumoniae, Neisseria gonorrhoeae and Staphylococcus aureus) and one fungal strain (Candida albicans). Eight of the plants inhibited the growth of all microorganisms at a concentration range of 0.4 mg/ml to 12.5 mg/ml. Acacia karroo showed the most promising antimicrobial activity, with a minimum inhibitory concentration (MIC) of 0.4 mg/ml on Staphylococcus aureus and 0.8 mg/ml on Neisseria gonorrhoeae. All ten plants were further investigated for their antioxidant activities using the DPPH scavenging method. Acacia karroo and Rhoicissus tridentata subsp. cuneifolia showed good antioxidant activity with IC50 values of 0.83 mg/ml and 0.06 mg/ml, respectively. The toxicity of plants was determined using the XTT reduction method against Vero cells. None of the ten plants showed toxicity on the cells. The obtained results confirmed that Acacia karroo and possibly Rhoicissus tridentata subsp. cuneifolia have the potential of being used as antimicrobial agents in the treatment of STDs and UTIs. These results support and validate traditional use of medicinal plants studied.

Keywords: antimicrobial, antioxidant, Neisseria gonorrhoeae, sexually transmitted diseases

Procedia PDF Downloads 318

Authors: Pankaj K. Jha, Ajeet K. Jha, Pravin Mishra

Abstract:

Goat raising is a popular livestock sub-commodity of mixed farming system in Nepal. Besides food and nutritional security, it has an important role in the economy of many peoples. Goat breeding through AI is commonly practiced worldwide. It is a very basic tool to speed up genetic improvement and increase productivity. For the goat genetic improvement program, the government of Nepal has imported some specialized exotic goat breeds and semen. Some progress has been made in the initiation of selective breeding within the local breeds and practice of AI with imported semen. Importance of AI in goats has drawn more attention among goat farmers. However, importing semen is not a permanent solution at national level; rather, it is more important to develop and establish its own frozen semen production technique. Semen quality and its relationship with fertility are said to be a major concern in animal production, hence accurate measurement of semen fertilizing potential is of great importance. The survivability of sperm cells depends on semen quality. Survivability of sperm cells is assessed through visual and microscopic evaluation of spermatozoal progressive motility and morphology. In Nepal, there is lack of scientific information on seminal attributes of buck semen, its dilution, cooling and freezing technique under management conditions of Nepal. Therefore, the objective of this review was to provide brief information about breeding system, semen production and artificial insemination in Nepalese goat.

Keywords: artificial insemination, goat, Nepal, semen

Procedia PDF Downloads 190

Authors: Rabah Iratni, Husain El Hasasna, Khawlah Athamneh, Halima Al Sameri, Nehla Benhalilou, Asma Al Rashedi

Abstract:

Background: Cancer therapies have witnessed great advances in the recent past, however, cancer continues to be a leading cause of death, with colorectal cancer being the fourth cause of cancer-related deaths. Colorectal cancer affects both sexes equally with poor survival rate once it metastasizes. Phytochemicals, which are plant derived compounds, have been on a steady rise as anti-cancer drugs due to the accumulation of evidences that support their potential. Here, we investigated the anticancer effect of Rhus coriaria on colon cancer cells. Material and Method: Human colon cancer HT-29 cell line was used. Protein expression and protein phosphorylation were examined using Western blotting. Transcription activity was measure using Quantitative RT-PCR. Human tumoral clonogenic assay was used to assess cell survival. Senescence was assessed by the senescence-associated beta-galactosidase assay. Results: Rhus coriaria extract (RCE) was found to significantly inhibit the viability and colony growth of human HT-29 colon cancer cells. RCE induced senescence and cell cycle arrest at G1 phase. These changes were concomitant with upregulation of p21, p16, downregulation of cyclin D1, p27, c-myc and expression of Senescence-associated-β-Galactosidase activity. Moreover, RCE induced non-canonical beclin-1independent autophagy and subsequent apoptotic cell death through activation of activation caspase 8 and caspase 7. The blocking of autophagy by 3-methyladenine (3-MA) or chloroquine (CQ) reduced RCE-induced cell death. Further, RCE induced DNA damage, reduced mutant p53 protein level and downregulated phospho-AKT and phospho-mTOR, events that preceded autophagy. Mechanistically, we found that RCE inhibited the AKT and mTOR pathway, a regulator of autophagy, by promoting the proteasome-dependent degradation of both AKT and mTOR proteins. Conclusion: Our findings provide strong evidence that Rhus coriaria possesses strong anti-colon cancer activity through induction of senescence and autophagic cell death, making it a promising alternative or adjunct therapeutic candidate against colon cancer.

Keywords: autophagy, proteasome degradation, senescence, mTOR, apoptosis, Beclin-1

Procedia PDF Downloads 239

Authors: Kyriaki Hatziagapiou, Konstantinos Bethanis, Olti Nikola, Elias Christoforides, Eleni Koniari, Eleni Kakouri, George Lambrou, Christina Kanaka-Gantenbein

Abstract:

Glioblastoma multiforme (GBM) remains a serious health challenge, as current therapeutic modalities continue to yield unsatisfactory results, with the average survival rarely exceeding 1-2 years. Natural compounds still provide some of the most promising approaches for discovering new drugs. The non-psychotropic cannabidiol (CBD) deriving from Cannabis sativa L. provides such promise. CBD is endowed with anticancer, antioxidant, and genoprotective properties as established in vitro and in in vivo experiments. CBD’s selectivity towards cancer cells and its safe profile suggest its usage in cancer therapies. However, the bioavailability of oral CBD is low due to poor aqueous solubility, erratic gastrointestinal absorption, and significant first-pass metabolism, hampering its therapeutic potential and resulting in a variable pharmacokinetic profile. In this context, CBD can take great advantage of nanomedicine-based formulation strategies. Cyclodextrins (CDs) are cyclic oligosaccharides used in the pharmaceutical industry to incorporate apolar molecules inside their hydrophobic cavity, increasing their stability, water solubility, and bioavailability or decreasing their side effects. CBD-inclusion complexes with CDs could be a good strategy to improve its properties, like solubility and stability to harness its full therapeutic potential. The current research aims to study the potential cytotoxic effect of CBD and CBD-CDs complexes CBD-RMβCD (randomly methylated β-cyclodextrin) and CBD-HPβCD (hydroxypropyl-b-CD) on the A172 glioblastoma cell line. CBD is diluted in 10% DMSO, and CBD/CDs solutions are prepared by mixing solid CBD, solid CDs, and dH2O. For the biological assays, A172 cells are incubated at a range of concentrations of CBD, CBD-RMβCD and CBD-HPβCD, RMβCD, and HPβCD (0,03125-4 mg/ml) at 24, 48, and 72 hours. Analysis of cell viability after incubation with the compounds is performed with Alamar Blue viability assay. CBD’s dilution to DMSO 10% was inadequate, as crystals are observed; thus cytotoxicity experiments are not assessed. CBD’s solubility is enhanced in the presence of both CDs. CBD/CDs exert significant cytotoxicity in a dose and time-dependent manner (p < 0.005 for exposed cells to any concentration at 48, 72, and 96 hours versus cells not exposed); as their concentration and time of exposure increases, the reduction of resazurin to resofurin decreases, indicating a reduction in cell viability. The cytotoxic effect is more pronounced in cells exposed to CBD-HPβCD for all concentrations and time-points. RMβCD and HPβCD at the highest concentration of 4 mg/ml also exerted antitumor action per se since manifesting cell growth inhibition. The results of our study could afford the basis of research regarding the use of natural products and their inclusion complexes as anticancer agents and the shift to targeted therapy with higher efficacy and limited toxicity. Acknowledgments: The research is partly funded by ΙΚΥ (State Scholarships Foundation) – Post-doc Scholarships-Partnership Agreement 2014-2020.

Keywords: cannabidiol, cyclodextrins, glioblastoma, hydroxypropyl-b-Cyclodextrin, randomly-methylated-β-cyclodextrin

Procedia PDF Downloads 156

Authors: Fadilah Aleanizy

Abstract:

Colicins are a family of bacterial toxins that kill Escherichia coli and other closely related species. The mode of action of colicins involves binding to an outer membrane receptor and translocation across the cell envelope, leading to cytotoxicity through specific targets. The mechanism of colicin cytotoxicity includes a non-specific endonuclease activity or depolarization of the cytoplasmic membrane by pore-forming activity. For Group A colicins, translocation requires an interaction between the N-terminal domain of the colicin and a series of membrane- bound and periplasmic proteins known as the Tol system (TolB, TolR, TolA, TolQ, and Pal and the active domain must be translocated through the outer membranes. Protein-protein interactions are intrinsic to virtually every cellular process. The transient protein-protein interactions of the colicin include the interaction with much more complicated assemblies during colicin translocation across the cellular membrane to its target. The potassium release assay detects variation in the K+ content of bacterial cells (K+in). This assays is used to measure the effect of pore-forming colicins such as ColA on an indicator organism by measuring the changes of the K+ concentration in the external medium (K+out ) that are caused by cell killing with a K+ selective electrode. One of the goals of this work is to employ a quantifiable in-vivo method to spot which Tol protein are more implicated in the interaction with colicin A as it is translocated to its target.

Keywords: K+ efflux, Colicin A, Tol-proteins, E. coli

Procedia PDF Downloads 393

Authors: Bhumika Wadhwa, Fayaz Malik

Abstract:

The serine/threonine protein kinase B (PKB) also known as Akt, is one of the multifaceted kinase in human kinome, existing in three isoforms. Akt plays a vital role in phosphoinositide 3-kinase (PI3K) mediated oncogenesis in various malignancies and is one of the attractive targets for cancer drug discovery. The functional significance of an individual isoform of Akt is not redundant in cancer cell proliferation and metastasis instead Akt isoforms play distinct roles during metastasis; thereby regulating EMT. This study aims to determine isoform specific functions of Akt in cancer. The results obtained suggest that Akt1 restrict tumor invasion, whereas Akt2 promotes cell migration and invasion by various techniques like MTT, wound healing and invasion assay. Similarly, qRT-PCR also revealed that Akt3 has shown promising results in promoting cancer cell migration. Contrary to pro-oncogenic properties attributed to Akt, it is to be understood how various isoforms of Akt compensates each other in the regulation of common pathways during cancer progression and drug resistance. In conclusion, this study aims to target selective isoforms which is essential to inhibit cancer. However, the question now is whether, and how much, Akt inhibition will be tolerated in the clinic remains to be answered and the experiments will have to address the question of which combinations of newly devised Akt isoform specific inhibitors exert a favourable therapeutic effect in in vivo models of cancer to provide the therapeutic window with minimal toxicity.

Keywords: Akt isoforms, cancer, drug resistance, epithelial mesenchymal transition

Procedia PDF Downloads 239

Authors: Nafiseh Mollaei, Reihaneh Kardehi Moghaddam

Abstract:

In case of Diabetes Mellitus the controlling of insulin is very difficult. This illness is an incurable disease affecting millions of people worldwide. Glucose is a sugar which provides energy to the cells. Insulin is a hormone which supports the absorption of glucose. Fuzzy control strategy is attractive for glucose control because it mimics the first and second phase responses that the pancreas beta cells use to control glucose. We propose two control algorithms a type-1 fuzzy controller and an interval type-2 fuzzy method for the insulin infusion. The closed loop system has been simulated for different patients with different parameters, in present of the food intake disturbance and it has been shown that the blood glucose concentrations at a normoglycemic level of 110 mg/dl in the reasonable amount of time. This paper deals with type 1 diabetes as a nonlinear model, which has been simulated in MATLAB-SIMULINK environment. The novel model, termed the Augmented Minimal Model is used in the simulations. There are some uncertainties in this model due to factors such as blood glucose, daily meals or sudden stress. In addition to eliminate the effects of uncertainty, different control methods may be utilized. In this article, fuzzy controller performance were assessed in terms of its ability to track a normoglycemic set point (110 mg/dl) in response to a [0-10] g meal disturbance. Finally, the development reported in this paper is supposed to simplify the insulin delivery, so increasing the quality of life of the patient.

Keywords: interval type-2, fuzzy controller, minimal augmented model, uncertainty

Procedia PDF Downloads 413

Authors: Zhang Zhenzhen

Abstract:

Valproate (VPA) is commonly used in the treatment of bipolar disorder and epilepsy. The mechanism is complicated, including its ability to inhibit histone deacetylases (HDACs). Here, we show that VPA attenuated VEGF gene expression and the morphological changes in choroidal neovascularization (CNV) induced by photocoagulation in retina. C57BL/6 mice were injected subcutaneously at 300mg/kg twice daily with VPA before insult. Vascular endothelial growth factor (VEGF)-A and VEGF-B were examined in the eyes of VPA-treated mice and in human retinal pigment epithelial cell lines (ARPE-19) exposed to VPA. In addition, CNV was induced by photocoagulation in mice injected with VPA, and the volume of CNV was compared by fluorescence-labeled choroidal flat mount. Morphological changes were analyzed on stained histological sections. Western blot analysis was used to determine protein levels of VEGF-A and VEGF-B, and acetylation of histone H3 in each group. VPA injected intraperitoneally attenuated the VEGF-A and VEGF-B expression in the retina, accompanied by the hyperacetylation of retina tissue, indicating that VPA acts directly on retina tissues through acetylation to reduce the expression of VEGF. VPA also attenuated the VEGF-A mRNA expression in the retinal pigment epithelium showed by immunohistochemistry. Moreover, the administration of VPA significantly attenuated photocoagulation-induced CNV in mice. These results demonstrate that VPA attenuated VEGF production in retina associated with choroidal neovascularization possibly via the HDAC inhibition.

Keywords: retina, acetylation, chorodial neovascularization, vascular endothelial growth factor

Procedia PDF Downloads 178

Authors: Emmanuel Billy

Abstract:

This work is in the CABRISS project (H2020 projects) which aims at developing innovative cost-effective methods for the extraction of materials from the different sources of PV waste: Si based panels, thin film panels or Si water diluted slurries. Aluminum, silicon, indium, and silver will especially be extracted from these wastes in order to constitute materials feedstock which can be used later in a closed-loop process. The extraction of metals from silicon solar cells is often an energy-intensive process. It requires either smelting or leaching at elevated temperature, or the use of large quantities of strong acids or bases that require energy to produce. The energy input equates to a significant cost and an associated CO2 footprint, both of which it would be desirable to reduce. Thus there is a need to develop more energy-efficient and environmentally-compatible processes. Thus, ‘ionometallurgy’ could offer a new set of environmentally-benign process for metallurgy. This work demonstrates that ionic liquids provide one such method since they can be used to dissolve and recover silver. The overall process associates leaching, recovery and the possibility to re-use the solution in closed-loop process. This study aims to evaluate and compare different ionic liquids to leach and recover silver. An electrochemical analysis is first implemented to define the best system for the Ag dissolution. Effects of temperature, concentration and oxidizing agent are evaluated by this approach. Further, a comparative study between conventional approach (nitric acid, thiourea) and the ionic liquids (Cu and Al) focused on the leaching efficiency is conducted. A specific attention has been paid to the selection of the Ionic Liquids. Electrolytes composed of chelating anions are used to facilitate the lixiviation (Cl, Br, I,), avoid problems dealing with solubility issues of metallic species and of classical additional ligands. This approach reduces the cost of the process and facilitates the re-use of the leaching medium. To define the most suitable ionic liquids, electrochemical experiments have been carried out to evaluate the oxidation potential of silver include in the crystalline solar cells. Then, chemical dissolution of metals for crystalline solar cells have been performed for the most promising ionic liquids. After the chemical dissolution, electrodeposition has been performed to recover silver under a metallic form.

Keywords: electrodeposition, ionometallurgy, leaching, recycling, silver

Procedia PDF Downloads 229

Authors: Yilei Xue, Yat-Hing Ham, Wenting Qiu, Wan Chan, Stefan Nagl

Abstract:

Polydopamine (PDA) is a popular material in biological and medical applications due to its excellent biocompatibility, outstanding physicochemical properties, and facile fabrication. In this project, a new sandwich-structured PDA and silver (Ag) hybrid material named PDA-Ag-PDA was synthesized and characterized layer-by-layer, where silver nanoparticles (Ag NPs) are wrapped in PDA coatings, using SEM, AFM, 3D surface metrology, and contact angle meter. The silver loading capacity is positively proportional to the roughness value of the initial PDA film. This designed film was subsequently integrated within a digital microfluidic (DMF) platform coupling with an oxygen sensor layer for on-chip antibacterial assay. The concentration of E. coli was quantified on DMF by real-time monitoring oxygen consumption during E. coli growth with the optical oxygen sensor layer. The PDA-Ag-PDA coating shows an 99.9% reduction in E. coli population under non-nutritive condition with 1-hour treatment and has a strong growth inhibition of E. coliin nutrient LB broth as well. Furthermore, PDA-Ag-PDA film maintaining a low cytotoxicity effect to human cells. After treating with PDA-Ag-PDA film for 24 hours, 82% HEK 293 and 86% HeLa cells were viable. The SERS enhancement factor of PDA-Ag-PDA is estimated to be 1.9 × 104 using Rhodamine 6G (R6G). Multifunctional PDA-Ag-PDA coating provides an alternative platform to conjugate biomolecules and perform biological applications on DMF, in particular, for the adhesive protein and cell study.

Keywords: polydopamine, silver nanoparticles, digital microfluidic, optical sensor, antimicrobial assay, SERS

Procedia PDF Downloads 81

Authors: Somnath Mahato, Minarul Islam Sarkar, Luis Guillermo Gerling, Joaquim Puigdollers, Asit Kumar Kar

Abstract:

Nanostructured Zinc Oxide (ZnO) thin films have been successfully deposited on indium tin oxide (ITO) coated glass substrates by a simple two electrode electrodeposition process at constant potential. The preparative parameters such as deposition time, deposition potential, concentration of solution, bath temperature and pH value of electrolyte have been optimized for deposition of uniform ZnO thin films. X-ray diffraction studies reveal that the prepared ZnO thin films have a high preferential oriented c-axis orientation with compact hexagonal (wurtzite) structure. Surface morphological studies show that the ZnO films are smooth, continuous, uniform without cracks or holes and compact with nanorod-like structure on the top of the surface. Optical properties reveal that films exhibit higher absorbance in the violet region of the optical spectrum; it gradually decreased in the visible range with increases in wavelength and became least at the beginning of NIR region. The photoluminescence spectra shows that the observed peaks are attributed to the various structural defects in the nanostructured ZnO crystal. The microstructural and optical properties suggest that the electrodeposited ZnO thin films are suitable for application in photosensitive devices such as photovoltaic solar cells photoelectrochemical cells and light emitting diodes etc.

Keywords: electrodeposition, microstructure, optical properties, ZnO thin films

Procedia PDF Downloads 301

Authors: Juan Jose Filgueira, Daniela Londono-Serna, Liliana Maria Hoyos

Abstract:

Carnation (Dianthus caryophyllus) is one of the most important products of exportation in the floriculture industry worldwide. Fusariosis is the disease that causes the highest losses on farms, in particular the one produced by Fusarium oxysporum f.sp. dianthi, called vascular wilt. Gene identification and metabolic routes of the genes that participate in the building of the plant response to Fusarium are some of the current targets in the carnation breeding industry. The techniques for the identifying of resistant genes in the plants, is the analysis of the transcriptome obtained during the host-pathogen interaction. In this work, we report the cell transcriptome of different varieties of carnation that present differential response from Fusarium oxysporum f.sp. dianthi attack. The cells of the different hybrids produced in the outbreeding program were cultured in vitro and elicited with the parasite in a dual culture. The isolation and purification of mRNA was achieved by using affinity chromatography Oligo dT columns and the transcriptomes were obtained by using Illumina NGS techniques. A total of 85,669 unigenes were detected in all the transcriptomes analyzed and 31,000 annotations were found in databases, which correspond to 36.2%. The library construction of genic expression techniques used, allowed to recognize the variation in the expression of genes such as Germin-like protein, Glycosyl hydrolase family and Cinnamate 4-hydroxylase. These have been reported in this study for the first time as part of the response mechanism to the presence of Fusarium oxysporum.

Keywords: Carnation, Fusarium, vascular wilt, transcriptome

Procedia PDF Downloads 129

Authors: S. Ilahi, S. Almosni, F. Chouchene, M. Perrin, K. Zelazna, N. Yacoubi, R. Kudraweic, P. Rale, L. Lombez, J. F. Guillemoles, O. Durand, C. Cornet

Abstract:

Great efforts have been dedicated to obtain high quality of GaAsPN. The properties of GaAsPN have played a great part on the development of solar cells devices based in Si substrate. The incorporation of N in GaAsPN that having a band gap around of 1.7 eV is of special interest in view of growing in Si substrate. In fact, post-growth and rapid thermal annealing (RTA) could be an effective way to improve the quality of the layer. Then, the influence of growth conditions and post-growth annealing on optical and thermal parameters is considered. We have used Photothermal deflection spectroscopy PDS to investigate the impact of rapid thermal annealing on thermal and optical properties of GaAsPN. In fact, the principle of the PDS consists to illuminate the sample by a modulated monochromatic light beam. Then, the absorbed energy is converted into heat through the nonradiative recombination process. The generated thermal wave propagates into the sample and surrounding media creating a refractive-index gradient giving rise to the deflection of a laser probe beam skimming the sample surface. The incident light is assumed to be uniform, and only the sample absorbs the light. In conclusion, the results are promising revealing an improvement in absorption coefficient and thermal conductivity.

Keywords: GaAsPN absorber, photothermal defelction technique PDS, photonics on silicon, thermal conductivity

Procedia PDF Downloads 340

Authors: Saifullah K., Muhammad Naziruddin Saifullah, Muhammad N.

Abstract:

The bio control potential and mechanism of P. chlamydosporia against Meloidogyne incognita was evaluated in the present study. Under invitro conditions, P. chlamydosporia was tested for parasitism of eggs and females of M. incognita. The results indicated that this fungus parasitized 87% eggs and 82% females. Culture filtrate (CF) of P. chlamydosporia was tested for its larvicide activity against M. incognita 2nd stage juvenile. The maximum mortality was 97.3% at 100% concentration of the culture filtrate while minimum mortality was 7.3% in 25% concentration after 24 hrs. The result of the pot experiment proved that P. chlamydosporia has reduced the incidence of RKN and improved all tested agronomic growth parameters. The treatment with inoculated M. incognita alone reduced plant height, fresh shoot, and fresh root weight by 44.7%, 29.8%, and 32.8% respectively over uninoculated healthy control. Histopathological studies on the interaction of Pochonia chlamydosporia and Meloidogyne incognita on tomato roots revealed anatomical changes among treatments. Less number of galls with small in size and scarcer abnormalities in the vascular cylinder was observed in plants inoculated with P. chlamydosporia and M. incognita than the plants treated with nematode only. The fungus was seen in in the intercellular spaces of cortical and epidermal cells while the vascular bundles of the plant remain intact, inoculated only with P. chlamydosporia. In the infected roots, many mature females were seen which feed on giant cells. The findings also revealed that control healthy plants were not affected and no histological changes were noted.

Keywords: histopathology, Pochonia chlamydosporia, Meloidogyne incognita, tomato

Procedia PDF Downloads 87

Authors: Mohamed H. El-Newehy, Badr M. Thamer, Nasser A. M. Barakat, Mohammad A.Abdelkareem, Salem S. Al-Deyab, Hak Y. Kim

Abstract:

In this study, the influence of nitrogen doping on the electrocatalytic activity of carbon nanofibers with nickel nanoparticles toward methanol oxidation is introduced. The modified carbon nanofibers have been synthesized from calcination of electrospun nanofiber mats composed of nickel acetate tetrahydrate, poly(vinyl alcohol) and urea in argon atmosphere at 750oC. The utilized physicochemical characterizations indicated that the proposed strategy leads to form carbon nanofibers having nickel nanoparticles and doped by nitrogen. Moreover, due to the high-applied voltage during the electrospinning process, the utilized urea chemically bonds with the polymer matrix, which leads to form nitrogen-doped CNFs after the calcination process. Investigation of the electrocatalytic activity indicated that nitrogen doping NiCNFs strongly enhances the oxidation process of methanol as the current density increases from 52.5 to 198.5 mA/cm2 when the urea content in the original electrospun solution was 4 wt% urea. Moreover, the nanofibrous morphology exhibits distinct impact on the electrocatalytic activity. Also, nitrogen-doping enhanced the stability of the introduced Ni-based electrocatalyst. Overall, the present study introduces effective and simple strategy to modify the electrocatalytic activity of the nickel-based materials.

Keywords: electrospinning, methanol electrooxidation, fuel cells, nitrogen-doping, nickel

Procedia PDF Downloads 413

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

Abstract:

The deer velvet antler is well known for its traditional medicinal value and is widely used in the clinic. It has been considered to possess bone-strengthening activity. The goal of this study was to investigate the anti-osteoporotic effect of deer antler velvet on ovariectomized rats (OVX), and their possible mechanism of the action. In the first step, the in vitro effects of DAE on bone loss were determined. The proliferation, collagen content and alkaline phosphatase (ALP) activity of human osteoblastic MG-63 cells and osteoclastogenesis from bone marrow-derived precursor cells were measured. The in vivo experiment confirmed the positive effect of DAE on bone tissue. 3-month old female Sparague-Dawley rats were either sham operated or OVX, and administered DAE (20 and 100 mg/kg) for 4 weeks. DAE increased MG-63 cell proliferation and ALP activity in a dose-dependent manner. Collagen content was also increased by DAE treatment. However, the effect of DAE on bone resorption was not observed. OVX rats supplemented with DAE showed osteoprotective effects as the bone ALP level was increased and c-terminal telopeptide level was decreased by 100 mg/kg DAE treatment compared with OVX controls. Moreover, the tartrate-resistant acid phosphatase-5b level was also decreased by DAE treatment. The present study suggests that DAE is effective in preventing bone loss in OVX rats, and may be potential therapeutic agents for the treatment of postmenopausal osteoporosis.

Keywords: bone ALP, c-terminal telopeptide, deer antler, osteoporosis, ovariectomy, tartrate-resistant acid phosphatase-5b

Procedia PDF Downloads 232

Authors: Tauseef Ahmad, Muhammad Ishaq, Mathew Eapen, Ahyoung Park, Sam Karpiniec, Vanni Caruso, Rajaraman Eri

Abstract:

Fucoidans are complex, fucose-rich sulfated polymers discovered in brown seaweeds. Fucoidans are popular around the world, particularly in the nutraceutical and pharmaceutical industries, due to their promising medicinal properties. Fucoidans have been shown to have a variety of biological activities, including anti-inflammatory effects. They are known to inhibit inflammatory processes through a variety of mechanisms, including enzyme inhibition and selectin blockade. Inflammation is a part of the complicated biological response of living systems to damaging stimuli, and it plays a role in the pathogenesis of a variety of disorders, including arthritis, inflammatory bowel disease, cancer, and allergies. In the current investigation, various fucoidan extracts from Undaria pinnatifida, Fucus vesiculosus, Macrocystis pyrifera, Ascophyllum nodosum, and Laminaria japonica were assessed for inhibition of pro-inflammatory cytokine production (TNF-α, IL-1β, and IL-6) in LPS induced human macrophage cell line (THP-1) and human peripheral blood mononuclear cells (PBMCs). Furthermore, we also sought to catalogue these extracts based on their anti-inflammatory effects in the current in-vitro cell model. Materials and Methods: To assess the cytotoxicity of fucoidan extracts, MTT (3-[4,5-dimethylthiazol-2-yl]-2,5, -diphenyltetrazolium bromide) cell viability assay was performed. Furthermore, a dose-response for fucoidan extracts was performed in LPS induced THP-1 cells and PBMCs after pre-treatment for 24 hours, and levels of TNF-α, IL-1β, and IL-6 cytokines were measured using Enzyme-Linked Immunosorbent Assay (ELISA). Results: The MTT cell viability assay demonstrated that fucoidan extracts exhibited no evidence of cytotoxicity in THP-1 cells or PBMCs after 48 hours of incubation. The results of the sandwich ELISA revealed that all fucoidan extracts suppressed cytokine production in LPS-stimulated PBMCs and human THP-1 cells in a dose-dependent manner. Notably, at lower concentrations, the lower molecular fucoidan (5-30 kDa) extract from Macrocystis pyrifera was a highly efficient inhibitor of pro-inflammatory cytokines. Fucoidan extracts from all species including Undaria pinnatifida, Fucus vesiculosus, Macrocystis pyrifera, Ascophyllum nodosum, and Laminaria japonica exhibited significant anti-inflammatory effects. These findings on several fucoidan extracts provide insight into strategies for improving their efficacy against inflammation-related diseases. Conclusion: In the current research, we have successfully catalogued several fucoidan extracts based on their efficiency in LPS-induced macrophages and PBMCs in downregulating the key pro-inflammatory cytokines (TNF-, IL-1 and IL-6), which are prospective targets in human inflammatory illnesses. Further research would provide more information on the mechanism of action, allowing it to be tested for therapeutic purposes as an anti-inflammatory medication.

Keywords: fucoidan, PBMCs, THP-1, TNF-α, IL-1β, IL-6, inflammation

Procedia PDF Downloads 44

Authors: Tayyaba Bari, Muhammad Hamza Anjum, Samra Kanwal, Fakhera Ikram

Abstract:

Osteoarthritis, a degenerative condition, affects more than 213 million individuals globally. Since articular cartilage has no or limited vessels, therefore, after deteriorating, it is unable to rejuvenate. Traditional approaches for cartilage repair, like autologous chondrocyte implantation, microfracture and cartilage transplantation are often associated with postoperative complications and lead to further degradation. Decellularized human umbilical cord has gained interest as a viable treatment for cartilage repair. Decellularization removes all cellular contents as well as debris, leaving a biologically active 3D network known as extracellular matrix (ECM). This matrix is biodegradable, non-immunogenic and provides a microenvironment for homeostasis, growth and repair. UC derived bioink function as 3D scaffolding material, not only mediates cell-matrix interactions but also adherence, proliferation and propagation of cells for 3D organoids. This study comprises different physical, chemical and biological approaches to optimize the decellularization of human umbilical cord (UC) tissues followed by the solubilization of these tissues to bioink formation. The decellularization process consisted of two cycles of freeze thaw where the umbilical cord at -20˚C was thawed at room temperature followed by dissection in small sections from 0.5 to 1cm. Similarly decellularization with ionic and non-ionic detergents Sodium dodecyl sulfate (SDS) and Triton-X 100 revealed that both concentrations of SDS i.e 0.1% and 1% were effective in complete removal of cells from the small UC tissues. The results of decellularization was further confirmed by running them on 1% agarose gel. Histological analysis revealed the efficacy of decellularization, which involves paraffin embedded samples of 4μm processed for Hematoxylin-eosin-safran and 4,6-diamidino-2-phenylindole (DAPI). ECM preservation was confirmed by Alcian Blue, and Masson’s trichrome staining on consecutive sections and images were obtained. Sulfated GAG’s content were determined by 1,9-dimethyl-methylene blue (DMMB) assay, similarly collagen quantification was done by hydroxy proline assay. This 3D bioengineered scaffold will provide a typical atmosphere as in the extracellular matrix of the tissue, which would be seeded with the mesenchymal cells to generate the desired 3D ink for in vitro and in vivo cartilage regeneration applications.

Keywords: umbilical cord, 3d printing, bioink, tissue engineering, cartilage regeneration

Procedia PDF Downloads 77

Authors: Nada A. El-Shahaw, Anas A.Salem, M. Kobeisy, Hoda M. Shabaan

Abstract:

Reactive oxygen species (ROS) is collective term both oxygen radical, such superoxide (O₂•), hydroxyl(OH•), peroxyl (RO₂), and hydroperoxyl (HO₂•), and certain non-radical oxidizing agents, such as hydrogen peroxide (H₂O₂), hypochlorous acid (HOCL), and ozone (O₃), that can be convert easily to radical. The importance of antioxidants is shown here punicalagin. Punicalagin is preventing the harmful effect of (ROS) in all cells, specially gonadal cells. So, the aim of study was to investigate effects of punicalagin (PL) on maternal live body weight (MLBW), number of services/conception (NS), conception rate (CR), gestation length (GL), kindling rate (KR), total litter size (TLS), live litter size (LLS), kit weight (KW), progesterone (P4) and estradiol-17 (E2) concentrations at 1st and 2nd pregnancy of young does. A total of 28 healthy virgin does (6 months old) were divided into 2 equal groups. Group I, each doe, was injected IM with 100 ug PL twice/week pre-mating and one time 3 days post-mating. Group II, each doe was injected IM with sterilized water (control). Blood samples were taken at pre-mating, mating, post-mating, throughout pregnancy, and immediately post-kindling for assaying P4 and E2. All does were naturally mated with fertile bucks. Results revealed that PL displayed their significant impacts on MLBW, NS/conception, CR, GL, KR, TLS, LLS, KWs (birth and weaning), P4 and E2 concentrations either at 1ˢᵗ/2ⁿᵈ pregnancy or both of them. Conclusively, PL improved pregnancy outcomes of young do particularly at 2ⁿᵈ pregnancy and could be recommended in rabbit's farms.

Keywords: punicalagin, pregnancy, estradiol-17β, progesterone, does

Procedia PDF Downloads 96

Authors: Isalira Peroba Ramos, Cherley Borba Vieira de Andrade, Grazielle Suhett, Camila Salata, Paulo Cesar Canary, Guilherme Visconde Brasil, Antonio Carlos Campos de Carvalho, Regina Coeli dos Santos Goldenberg

Abstract:

Background: The therapeutic options for patients with cancer now include increasingly complex combinations of medications, radiation therapy (RT), and surgical intervention. Many of these treatments have important potential adverse cardiac effects and are likely to have significant effects on patient outcomes. Cell therapy appears to be promising for the treatment of chronic and degenerative diseases, including cardiomyopathy induced by RT, as the current therapeutic options are insufficient. Aims: To evaluate the potential of bone marrow mesenchymal cells (BMMCs) in radioinduced cardiac damage Methods: Female Wistar rats, 3 months old (Ethics Committee 054/14), were divided into 2 groups, non-treated irradiated group (IR n=15) and irradiated and BMMC treated (IRT n=10). Echocardiography was performed to evaluate heart function. After euthanasia, 3 months post treatment; the left ventricle was removed and prepared for RT-qPCR (VEGF and Pro Collagen I) and histological (picrosirius) analysis. Results: In both groups, 45 days after irradiation, ejection fraction (EF) was in the normal range for these animals (> 70%). However, the BMMC treated group had EF (83.1%±2.6) while the non-treated IR group showed a significant reduction (76.1%±2.6) in relation to the treated group. In addition, we observed an increase in VEGF gene expression and a decrease in Pro Collagen I in IRT when compared to IR group. We also observed by histology that the collagen deposition was reduced in IRT (10.26%±0.83) when compared to IR group (25.29%±0.96). Conclusions: Treatment with BMMCs was able to prevent ejection fraction reduction and collagen deposition in irradiated animals. The increase of VEGF and the decrease of pro collagen I gene expression might explain, at least in part, the cell therapy benefits. All authors disclose no financial or personal relationships with individuals or organizations that could be perceived to bias their work. Sources of funding: FAPERJ, CAPES, CNPq, MCT.

Keywords: mesenchymal cells, radioation, cardiotoxicity, bone marrow

Procedia PDF Downloads 238

Authors: Amina Ben Abla, Sylvie Changotade, Geraldine Rohman, Guilhem Boeuf, Cyrine Dridi, Ahmed Elmarjou, Florence Dufour, Didier Lutomski, Abdellatif Elm’semi

Abstract:

Understanding cell adhesion and interaction with the extracellular matrix is essential for biomedical and biotechnological applications, including the development of biomaterials. In recent years, numerous biomaterials have emerged and were used in the field of tissue engineering. Nevertheless, the lack of interaction of biomaterials with cells still limits their bio-integration. Thus, the design of bioactive biomaterials to improve cell attachment and proliferation is of growing interest. In this study, bio-functionalized material was developed combining a synthetic polymer scaffold surface with selected domains of type III human fibronectin (FNIII-DOM) to promote cell adhesion and proliferation. Bioadhesive ligand includes cell-binding domains of human fibronectin, a major ECM protein that interacts with a variety of integrins cell-surface receptors, and ECM proteins through specific binding domains were engineered. FNIII-DOM was produced in bacterial system E. coli in 5L fermentor with a high yield level reaching 20mg/L. Bioactivity of the produced fragment was validated by studying cellular adhesion of human cells. The adsorption and immobilization of FNIII-DOM onto the polymer scaffold were evaluated in order to develop an innovative biomaterial.

Keywords: biomaterials, cellular adhesion, fibronectin, tissue engineering

Procedia PDF Downloads 130

Authors: Laura Gantley, Vanessa M. Conn, Stuart Webb, Kirsty Kirk, Marta Gabryelska, Duncan Holds, Brett W. Stringer, Simon J. Conn

Abstract:

Trinucleotide repeat disorders comprise ~20 severe, inherited human neuromuscular and neurodegenerative disorders, which are a result of an abnormal expansion of repetitive sequences in the DNA. The most common of these, Huntington’s disease, results from the expansion of the CAG repeat region in exon 1 of the HTT gene via an unknown mechanism. Non-coding RNAs have been implicated in the initiation and progression of many diseases; thus, we focus on one circular RNA (circRNA) molecule arising from non-canonical splicing (back splicing) of HTT pre-mRNA. This circRNA and its mouse orthologue were transgenically overexpressed in human cells (SHSY-5Y and HEK293T) and mouse cells (Mb1), respectively. High-content imaging and flow cytometry demonstrated the overexpression of this circRNA reduces cell proliferation, reduces nuclear size independent of cellular size, and alters cell cycle progression. Analysis of protein by western blot and immunofluorescence demonstrated no change to HTT protein levels but altered nuclear-cytoplasmic distribution without impacting the expansion of the HTT repeat region. As these phenotypic and genotypic changes are found in Huntington’s disease patients, these results may suggest that this circRNA may play a functional role in the progression of Huntington’s disease.

Keywords: cell biology, circular RNAs, Huntington’s disease, molecular biology, neurodegenerative disorders

Procedia PDF Downloads 83

Authors: Miaomiao Zhang, Sabrina Beckmann, Haluk Ertan, Rocky Chau, Mike Manefield

Abstract:

Phenazines are a large class of nitrogen-containing aromatic heterocyclic compounds, which are almost exclusively produced by bacteria from diverse genera including Pseudomonas and Streptomyces. Phenazine-1-carboxylic acid (PCA) as one of 'core' phenazines are converted from chorismic acid before modified to other phenazine derivatives in different cells. Phenazines have attracted enormous interests because of their multiple roles on biocontrol, bacterial interaction, biofilm formation and fitness of their producers. However, in spite of ecological importance, degradation as a part of phenazines’ fate only have extremely limited attention now. Here, to isolate PCA-degrading bacteria, 200 mg L-1 PCA was supplied as sole carbon, nitrogen and energy source in minimal mineral medium. Quantitative PCR and Reverse-transcript PCR were employed to study abundance and activity of functional gene MFORT 16269 in PCA degradation, respectively. Intermediates and products of PCA degradation were identified with LC-MS/MS. After enrichment and isolation, a PCA-degrading strain was selected from soil and was designated as Rhodanobacter sp. PCA2 based on full 16S rRNA sequencing. As determined by HPLC, strain PCA2 consumed 200 mg L-1 (836 µM) PCA at a rate of 17.4 µM h-1, accompanying with significant cells yield from 1.92 × 105 to 3.11 × 106 cells per mL. Strain PCA2 was capable of degrading other phenazines as well, including phenazine (4.27 µM h-1), pyocyanin (2.72 µM h-1), neutral red (1.30 µM h-1) and 1-hydroxyphenazine (0.55 µM h-1). Moreover, during the incubation, transcript copies of MFORT 16269 gene increased significantly from 2.13 × 106 to 8.82 × 107 copies mL-1, which was 2.77 times faster than that of the corresponding gene copy number (2.20 × 106 to 3.32 × 107 copies mL-1), indicating that MFORT 16269 gene was activated and played roles on PCA degradation. As analyzed by LC-MS/MS, decarboxylation from the ring structure was determined as the first step of PCA degradation, followed by cleavage of nitrogen-containing ring by dioxygenase which catalyzed phenazine to nitrosobenzene. Subsequently, phenylhydroxylamine was detected after incubation for two days and was then transferred to aniline and catechol. Additionally, genomic and proteomic analyses were also carried out for strain PCA2. Overall, the findings presented here showed that a newly isolated strain Rhodanobacter sp. PCA2 was capable of degrading phenazines through decarboxylation and cleavage of nitrogen-containing ring, during which MFORT 16269 gene was activated and played important roles.

Keywords: decarboxylation, MFORT16269 gene, phenazine-1-carboxylic acid degradation, Rhodanobacter sp. PCA2

Procedia PDF Downloads 207