Search results for: muscle stem cells

Authors: Narmeen Mehmood

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The current study was conducted to evaluate the pharmacognostic parameters, phytochemical analysis and antibacterial activity of the plant. Microscopic studies were carried out to determine various Pharmacognostic parameters. Section cutting of the leaf was also done. The study of the ariel parts of Beaumontia grandiflora resulted in the identification of fatty acids mixture and unsaponifiable matters. For the separation of various constituents of the plant, successive solvent extraction was carried out in a laboratory. Material and Methods: The study was carried out with all three extracts of Beaumontia grandiflora i.e. Petroleum ether, Chloroform and Methanol. For the separation of various constituents of the plant, successive solvent extraction was carried out in the laboratory. Raw data containing the measured zones of inhibition in mm was tabulated. Results: The microscopic studies showed the presence of Upper epidermis in surface view, Part of Lamina in section view, cortical parenchyma in longitudinal view, Parenchyma with collapsed tissues, Parenchyma Cells, Epidermal cells with a part of covering trichome, starch granules, reticulated thickened vessels, Transverse Section of leaf of Beaumontia grandiflora showed Upper Epidermis, Lower Epidermis, Hairs, Vascular Bundles, Parenchyma. Phytochemical analysis of leaves of Beaumontia grandiflora indicates that Alkaloids are present. There is a possibility of the presence of some bioactive components in the crude extracts due to which it shows strong activity. Petroleum ether extract shows a greater zone of inhibition at low concentrations. Conclusion: The alkaloids possess good antibacterial activity so the presence of alkaloids may be responsible for the antibacterial activity observed in the crude organic extract of Beaumontia grandiflora.

Keywords: successive solvent extraction, zone of inhibitions., microscopy, phytochemical analysis

Procedia PDF Downloads 27

Authors: Zhanara Suleimenova, Raushan Blieva, Aigerim Zhakipbekova, Inkar Tapenbayeva, Zhanar Narmuratova

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Green processes are based on innovative technologies that do not negatively affect the environment. Industrial enzymes originated from biological systems can effectively contribute to sustainable development through being isolated from microorganisms which are fermented using primarily renewable resources. Many widespread microorganisms secrete a significant amount of biocatalysts into the environment, which greatly facilitates the task of their isolation and purification. The ability to control the enzyme production through the regulation of their biosynthesis and the selection of nutrient media and cultivation conditions allows not only to increase the yield of enzymes but also to obtain enzymes with certain properties. In this regard, large potentialities are embedded in immobilized cells. Enzyme production technology in a secreted active form enabling industrial application on an economically feasible scale has been developed. This method is based on the immobilization of enzyme producers on a solid career. Immobilizing has a range of advantages: decreasing the price of the final product, absence of foreign substances, controlled process of enzyme-genesis, the ability of various enzymes' simultaneous production, etc. Design of proposed equipment gives the opportunity to increase the activity of immobilized cell culture filtrate comparing to free cells, growing in periodic culture conditions. Such technology allows giving a 10-times raise in culture productivity, to prolong the process of fungi cultivation and periods of active culture liquid generation. Also, it gives the way to improve the quality of filtrates (to make them more clear) and exclude time-consuming processes of recharging fermentative vials, that require manual removing of mycelium.

Keywords: industrial enzymes, immobilization, submerged fermentation, microscopic fungi

Procedia PDF Downloads 141

Authors: Gabriele Ciasca, Tanya E. Sassun, Eleonora Minelli, Manila Antonelli, Massimiliano Papi, Antonio Santoro, Felice Giangaspero, Roberto Delfini, Marco De Spirito

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Glioblastoma multiforme (GBM) is an extremely aggressive brain tumor, characterized by a diffuse infiltration of neoplastic cells into the brain parenchyma. Although rarely considered, mechanical cues play a key role in the infiltration process that is extensively mediated by the tumor microenvironment stiffness and, more in general, by the occurrence of aberrant interactions between neoplastic cells and the extracellular matrix (ECM). Here we provide a nano-mechanical characterization of the viscoelastic response of human GBM tissues by indentation-type atomic force microscopy. High-resolution elasticity maps show a large difference between the biomechanics of GBM tissues and the healthy peritumoral regions, opening possibilities to optimize the tumor resection area. Moreover, we unveil the nanomechanical signature of necrotic regions and anomalous vasculature, that are two major hallmarks useful for glioma staging. Actually, the morphological grading of GBM relies mainly on histopathological findings that make extensive use of qualitative parameters. Our findings have the potential to positively impact on the development of novel quantitative methods to assess the tumor grade, which can be used in combination with conventional histopathological examinations. In order to provide a more in-depth description of the role of mechanical cues in tumor progression, we compared the nano-mechanical fingerprint of GBM tissues with that of grade-I (WHO) meningioma, a benign lesion characterized by a completely different growth pathway with the respect to GBM, that, in turn hints at a completely different role of the biomechanical interactions.

Keywords: AFM, nano-mechanics, nanomedicine, brain tumors, glioblastoma

Procedia PDF Downloads 342

Authors: Le Xuan Cau

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Flash flood is occurred in short time rainfall interval: from 1 hour to 12 hours in small and medium basins. Flash floods typically have two characteristics: large water flow and big flow velocity. Flash flood is occurred at hill valley site (strip of lowland of terrain) in a catchment with large enough distribution area, steep basin slope, and heavy rainfall. The risk of flash floods is determined through Gridded Basin Flash Flood Potential Index (GBFFPI). Flash Flood Potential Index (FFPI) is determined through terrain slope flash flood index, soil erosion flash flood index, land cover flash floods index, land use flash flood index, rainfall flash flood index. Determining GBFFPI, each cell in a map can be considered as outlet of a water accumulation basin. GBFFPI of the cell is determined as basin average value of FFPI of the corresponding water accumulation basin. Based on GIS, a tool is developed to compute GBFFPI using ArcObjects SDK for .NET. The maps of GBFFPI are built in two types: GBFFPI including rainfall flash flood index (real time flash flood warning) or GBFFPI excluding rainfall flash flood index. GBFFPI Tool can be used to determine a high flash flood potential site in a large region as quick as possible. The GBFFPI is improved from conventional FFPI. The advantage of GBFFPI is that GBFFPI is taking into account the basin response (interaction of cells) and determines more true flash flood site (strip of lowland of terrain) while conventional FFPI is taking into account single cell and does not consider the interaction between cells. The GBFFPI Map of QuangNam, QuangNgai, DaNang, Hue is built and exported to Google Earth. The obtained map proves scientific basis of GBFFPI.

Keywords: ArcObjects SDK for NET, basin average value of FFPI, gridded basin flash flood potential index, GBFFPI map

Procedia PDF Downloads 381

Authors: Viktorija Voidogaitė

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Catholic education is based on the belief that every human being is created in the image and likeness of God. It is also influenced by the idea that the Kingdom of Heaven belongs to the humble and vulnerable. These principles emphasize the importance of serving the most vulnerable members of the Church community and promoting inclusivity without discrimination. This perspective emphasizes the need to protect the weakest members with compassion. However, realizing such an ideal in practice proves challenging, as the shortcomings and errors prevalent in any society often stem from the actions of Christians within that society. The evolution of these connections is observed throughout the historical development of Catholic education. In some European countries, Catholic education has become elitist, with limited room for inclusivity. This creates a conspicuous gap between the principles of the Evangelical community and elite Catholic schools and gymnasiums. Some schools appear to be most inclined to educate only those students who best align with their profile, leaving those needing assistance on the margins. As we advance into the third decade of the 21st century, there emerges a fundamental consideration: whether individuals who can assist the underprivileged and the infirm are being emphasized. Yet, it remains an open question whether these individuals will also possess the willingness and capability to construct a community or society that is inclusive and accessible to all.

Keywords: inclusion, Catholic education, inclusive education, becoming

Procedia PDF Downloads 66

Authors: Asha Bhardwaj, Pushpendu Biswas

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Carbon Dots (CDs) are known to be absorbing in the UV and emitting in the blue to visible region [1-2]. As CDs have high bio compatibility, high emission efficiency, are environment friendly and are nontoxic in nature, they are a material of great importance for various biomedical as well as optoelectronic applications [3-6]. For bioimaging and photothermal therapy in cancer treatment CDs should be highly photostable, show interaction with NIR band [3], show quick excretion from the body and have high Quantum yields. NIR I stands for emission in the 650-950 nm region and NIR II stands for emission in the 1000-1700 nm range [4]. Penetration depth of NIR II is larger than that of NIR-1 or visible light [5]. Also, it shows heating effect which is beneficial for selective ablation of cancer cells while not harming the healthy cells. Therefore, efforts are being made by the scientific community to synthesize CDs which emit in the NIR II region. Here we report CDs emitting in all the three regions (Visible, NIR I and NIR II) of electromagnetic spectra ranging from 300-1150 nm. Wide range emissive CDs and Fe doped CDs are prepared by a one-step hydrothermal method. Fe concentration has been increased in steps to assess the contribution from Fe incorporation in the CD lattice [6]. The CDs emit in three wavelength ranges, from 300-600 nm, 600-800 nm (NIR I) and 900 – 1150 nm (NIR II). Such kind of broad emission behaviour in single system carbon dots is being reported for the first time. Further excitation wavelength (λex) dependent emission characteristics reveal that the emission peak wavelength values are unaffected by the changing excitation wavelength in the visible region. Also, NIR I and NIR II emission is observed only for 300 and 310 nm excitation, hinting towards two photon and three photon emission [4]. Emission from 650- 1150 nm is not observed for λex > 310 nm. Additionally, as expected the absorption spectra also ranges from 250-600 nm, as compared to commonly observed blue or UV absorption in CDs. The exquisite ultra-wide range emitting nontoxic CDs can find application not only as broad band emitters but also in photothermal therapy for cancer cell theranostics.

Keywords: Broad Emission, absorption, Carbon dots, NIR Emission

Procedia PDF Downloads 9

Authors: Jamal A. Hassan, Ali Q. Abdulrazzaq, Sadiq J. Abass

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In this study, both the photoelastic, as well as the finite element methods, are used to study the stress distribution within human vertebra (L4) under forces similar to those that occur during normal life. Two & three dimensional models of vertebra were created by the software AutoCAD. The coordinates obtained were fed into a computer numerical control (CNC) tensile machine to fabricate the models from photoelastic sheets. Completed models were placed in a transmission polariscope and loaded with static force (up to 1500N). Stresses can be quantified and localized by counting the number of fringes. In both methods the Principle stresses were calculated at different regions. The results noticed that the maximum von-mises stress on the area of the extreme superior vertebral body surface and the facet surface with high normal stress (σ) and shear stress (τ). The facets and other posterior elements have a load-bearing function to help support the weight of the upper body and anything that it carries, and are also acted upon by spinal muscle forces. The numerical FE results have been compared with the experimental method using photoelasticity which shows good agreement between experimental and simulation results.

Keywords: photoelasticity, stress, load, finite element

Procedia PDF Downloads 286

Authors: Mengjie Qu, Yi Wang, Jiawei Ding, Siyu Chen, Yanan Di

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Marine ecosystem is facing intensified multiple stresses caused by environmental contaminants from human activities. Xenobiotics, such as benzo(α)pyrene (BaP) have been discharged into marine environment and cause hazardous impacts on both marine organisms and human beings. As a filter-feeder, marine mussels, Mytilus spp., has been extensively used to monitor the marine environment. However, their genomic alterations induced by such xenobiotics are still kept unknown. In the present study, gills, as the first defense barrier in mussels, were selected to evaluate the genetic instability alterations induced by the exposure to BaP both in vivo and in vitro. Both random amplified polymorphic DNA (RAPD) assay and comet assay were applied as the rapid tools to assess the environmental stresses due to their low money- and time-consumption. All mussels were identified to be the single species of Mytilus coruscus before used in BaP exposure at the concentration of 56 μg/l for 1 & 3 days (in vivo exposure) or 1 & 3 hours (in vitro). Both RAPD and comet assay results were showed significantly increased genomic instability with time-specific altering pattern. After the recovery period in 'in vivo' exposure, the genomic status was as same as control condition. However, the relative higher genomic instabilities were still observed in gill cells after the recovery from in vitro exposure condition. Different repair mechanisms or signaling pathway might be involved in the isolated gill cells in the comparison with intact tissues. The study provides the robust and rapid techniques to exam the genomic stability in marine organisms in response to marine environmental changes and provide basic information for further mechanism research in stress responses in marine organisms.

Keywords: genotoxic impacts, in vivo/vitro exposure, marine mussels, RAPD and comet assay

Procedia PDF Downloads 280

Authors: Anca Maria Cimpean, Serban Comsa

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Chick embryo chorioallantoic membrane (CAM) is a well vascularised in vivo experimental model used as a platform for testing the behavior of different implants inserted on it from tumor fragments to therapeutic agents or various biomaterials. Five types of collagen-based biomaterials with 2D and 3D structure (MotifMesh, Optimaix2D, Optimaix3D, Dual Layer Collagen and Xenoderm) were implanted on CAM and continuously evaluated by stereomicroscope for up to 5 days post-implant with an emphasis of their ability to requisite and develop new blood vessels (BVs) followed by microscopic analysis. MotifMEsh did not induce any angiogenic response lacking to be invaded by BVs from the CAM, but it induced intense inflammatory response necrosis and fibroblastic reaction around the implant. Optimaix2D has good adherence. CAM with minimal or no inflammatory reaction, a good integration of the CAM between the collagen mesh’s fibers, consistent adhesion of the cells to the collagen fibers,and a good ability to form pseudo-vascular channels filled with cells. Optimaix3D induced the highest angiogenic effects on CAM. The material shows good integration on CAM. The collagen fibers of the material show the ability to organize themselves into linear and tubular structures. It is possible to see blood elements, especially at the periphery of the implant. Dual-layer collagen behaves similar to Optimaix 3D, while Xenoderm induced a moderate angiogenic effect on CAM. Based on these data, we may conclude that collagen-based materials have variable ability to requisite and develop new blood vessels. A proper selection of collagen-based biomaterial scaffolds may crucially influence the acquisition and development of blood vessels during angiogenesis assays.

Keywords: chick embryo chorioallantoic membrane, collagen scaffolds, blood vessels, vascular microenvironment

Procedia PDF Downloads 195

Authors: Charn Loong Ng, Mamun Bin Ibne Reaz

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As elderly population grows globally, the percentage of people diagnosed with musculoskeletal disorder (MSD) increase proportionally. Electromyography (EMG) is an important biosignal that contributes to MSD’s clinical diagnose and recovery process. Conventional conductive electrode has many disadvantages in the continuous EMG measurement application. This research has design a new surface EMG biosensor based on the parallel-plate capacitive coupling principle. The biosensor is developed by using a double-sided PCB with having one side of the PCB use to construct high input impedance circuitry while the other side of the copper (CU) plate function as biosignal sensing metal plate. The metal plate is insulated using kapton tape for contactless application. The result implicates that capacitive biosensor is capable to constantly capture EMG signal without having galvanic contact to human skin surface. However, there are noticeable noise couple into the measured signal. Post signal processing is needed in order to present a clean and significant EMG signal. A complete design of single ended, non-contact, high input impedance, front end EMG biosensor is presented in this paper.

Keywords: contactless, capacitive, biosensor, electromyography

Procedia PDF Downloads 451

Authors: Ahsan Khan, Nazish Shah, Muhammad Salman

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The present study deals with the toxicological effects of copper, lead and chromium on brain and liver tissues of grass carp (Ctenopharyngodon idella). The average length of experimental fish was 8.5 ± 5.5 cm and weighed 9.5 ± 6.5 g. Grass carp was exposed to lethal concentration (LC₁₅) of copper, lead and chromium for 24, 48, 72 and 96 hours respectively. (LC₁₅) for copper was 1.5, 1.4, 1.2 and 1mgL⁻¹. Similarly, LC₁₅ of lead was 250, 235, 225 and 216mgL⁻¹ while (LC₁₅) for chromium was 25.5, 22.5, 20 and 18mgL⁻¹ respectively. During the time of exposure against various doses of heavy metals the grass carp showed some behavioral changes. In the initial stages of experiment, the rapid movements and gulping of air were observed. Several times the fish tried to jump to scat from the toxic median. In addition, the accumulation of heavy metals in different tissues of grass carp particularly in liver and brain tissues were observed. Lead was highly accumulated in brain tissue after the exposure of fish for 24 and 48 hours, while highly accumulated in liver tissues after the exposure of fish for 72 and 96 hours. Chromium was highly accumulated in the liver tissues after the exposure of fish for 24 hours while its accumulation was found highly in the brain tissues after the exposure of fish for 48, 72 and 96 hours. Similarly, accumulation of copper concentration was found highly in brain tissues after the exposure of 48 and 96 hours while its accumulation was high in liver tissues after the exposure of 24 and 72 hours. Comparatively maximum accumulation of lead was found in brain and liver tissues of grass carp followed by chromium and copper. Furthermore, accumulation of these metals caused many abnormalities like gliosis, destruction of cell, change in cell shape and shrinkage of cells in brain tissue while in liver tissues aggregation in hepatocytes, widen space between cells and also destruction of cell was observed. These experiments and observations can be useful to monitor the aquatic pollution and quality of aquatic environment system.

Keywords: brain, grass carp, liver, lethal concentration, toxicity

Procedia PDF Downloads 157

Authors: Aliaa M. Issa, Mahmoud N. ElRouby, Sahar A. S. Ahmad, Mahmoud M. El-Merzabani

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Sider honey is a type of honey produced by bees feeding on the nectar of Sider tree, Ziziphus spina-christi (L) Desf . Honey is an effective agent for preventing, inhibiting and treating the growth of human and animal cancer cell lines in vitro and in vivo. The aim of the present study was to evaluate the impact of different dilutions from crude Sider honey and different duration times of exposure on the growth of six tumor cell lines (human cervical cancer cell line, HeLa; human hepatocellular carcinoma cell line, HepG-2; human larynx carcinoma cell line, Hep-2; brain tumor cell line, U251) as well as one animal cancerous cell line (Ehrlich ascites carcinoma cells line, EAC) and one normal cell line, Homo sapiens, human, (WISH) CCL-25. Different concentrations and treatment durations with Sider honey were tested on the growth of several cancer cell lines types. Histopathological changes in the tumor masses, animal survival, apoptosis and necrosis of the used cancer cell lines (using flow cytometry) were evaluated. Sider honey was administers either to the tumor mass itself by intratumoral injection or via drinking water. One-way ANOVA test was used for the analysis of (the means + standard error) of the optical density obtained from the Elisa reader and flow cytometry. The study revealed that different concentrations of Sider honey affected the growth patterns of all the studied cancer cell lines as well as their histopathological changes, and it depended on the cell line nature and the concentration of honey used. It is obvious that the relative animal survival percentage (bearing Ehrlich ascites carcinoma, EAC cells) was proportionally increased with the increase in the used honey concentrations. The study of apoptosis and necrosis using the flow cytometry technique emphasized the viability results. In conclusion, Sider honey was effective as antitumor agent, in the used concentrations.

Keywords: antitumor, honey, sider, tumor cell lines

Procedia PDF Downloads 538

Authors: Meltem Ramoglu, Ertugrul Safran, Hikmet Ucgun, Busra Kepenek Varol, Hulya Nilgun Gurses

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Down’s syndrome (DS) is a human genetic disorder caused by the presence of all or part of an extra chromosome 21. Many patients with DS have musculoskeletal problems that affect weak muscle tone (hypotonia) and ligament laxity. This leads to excessive joint hypermobility and decreased position sense (proprioception). Lack of proprioception may cause balance problems. The aim of our study was to investigate how does joint hypermobility affect balance in patients with DS. Our study conducted with 13 DS patients age between 18 to 40 years. Demographic data were recorded. Beighton Hypermobility Score (BHS) was used to evaluate joint hypermobility. Balance score of participants was evaluated with Berg Balance Scale (BBS). Mean age of our participants was 29,8±3,57 year. Average score of body mass index and BHS were; 33,23 ±3,78 kg/m2 and 7,61±1,04, respectively. Out of a maximum possible score of 56 on the Berg Balance Scale, scores of participants with DS ranged from 36–51, with a mean of 43±4,45. Significant correlation was found between BHS and BBS (r: -,966, p=0.00). All of our participants have 6/9 or higher grade from BHS. As a conclusion of our study; joint hypermobility may affect balance score in patients with DS. The results suggest that people with DS have worse balance scores which affected by hypermobility. Further studies need larger population for more reliable results.

Keywords: adults, balance, Down's syndrome, joint hypermobility

Procedia PDF Downloads 331

Authors: Suneeta Gireesh Panicker

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Aim: Aminopeptidase P (APP) is an enzyme that has specificity for proline, can specifically cleave Xaa-Proline peptides and is a metallo-aminopeptidase. The bonds nearby to the imino acid proline are tough to cleave by many peptidases, but APP can specifically break peptide bonds engaged with proline. Membrane-bound form and a cytosolic form are the two forms in which this enzyme exists. The exact physiological function of APP remains unclear and hence the present work attempts to determine it. Methods: In the present study, the expression pattern of cytosolic Aminopeptidase P (DAP) was determined in all the embryonic stages and larval stages of wild-type Drosophila by using polyclonal monospecific antibodies. To show the presence of DAP RNA in embryonic and larval stages, RNA in situ hybridization was performed. DAP promoter-LacZ fusion reporter gene vector was used to construct transgenic embryos to study the regulation pattern of DAP. To study the DAP expression profile, a transgenic fly consisting of a DAP promoter with β-gal and GFP reporter genes in front of it was constructed. Results: DAP protein expression was observed in neuroectodermal cells, posterior midgut primordium, proctodeum, ventral neuroblast and primordial stomatogastric nervous system. It was observed in the ventral cord and midgut in stage 12. The completely developed embryos showed the intense occurrence of it in the ventral cord and gut region. The eye-antennal disc, wing disc and leg disc also showed the presence of DAP protein. LacZ expression in transgenic embryos also showed the same pattern. Conclusion: Similar to various known multiple-functional proteins, DAP could be one with different functions at different stages and in different cells. Data presented here designates DAP functions in the early embryonic and imaginal dics differentiation and development, suggesting that it may be required for the metabolism of proteins like neuropeptides and tachykinins.

Keywords: aminopeptidase P, in situ hybridization, transgenic fly, embryonic stages

Procedia PDF Downloads 87

Authors: V. Endo, A. G. Silva Sobrinho, F. A. Almeida, N. L. L. Lima, G. M. Manzi, L. G. A. Cirne, N. M. B. L. Zeola

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We used 24 Ile de France lambs, weighing between 15 and 32 kg (BW). Treatments were supplemented with concentrate: “in nature” sugarcane (IN), sugarcane hydrolyzed using 0.6% calcium oxide (CaO) under aerobic condition (AER), and sugarcane hydrolyzed using 0.6% CaO under anaerobic condition (ANA), constituting a completely randomized design with eight repetitions per treatment. Lambs were housed in individual stalls and fed into the through, allowing 10% of leftovers. Lambs were slaughtered when body weight reached 32 kg. The following parameters were determined on Longissimus lumborum muscle of hot and cold carcasses: pH and color, 45 minutes and 24 hours after slaughtering. Qualitative analysis of the meat were performed in the loins, water-holding capacity (WHC), cooking loss (CL), and shear force (SF). We used a completely randomized design with three treatments and eight repetitions. Means were compared by Tukey test at 5% significance. A higher value for redness (a*) 45 minutes after slaughter (10.48) was found for lambs fed sugarcane hydrolyzed under anaerobic conditions. The other qualitative characteristics of meat were not affected by treatments (P >0.05). The comparison of meat quality resulting from the treatments shows that it is possible to feed in nature sugarcane to lambs, thus waiving hydrolyses process and the spending with alkalizing agent.

Keywords: oxide, hydrolysis, meat quality, pH

Procedia PDF Downloads 565

Authors: Yuriko Takayama, Norihiro Kato

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Quorum sensing (QS) is a cell-to-cell communication system in bacteria to regulate expression of target genes. In gram-negative bacteria, activation on QS is controlled by a concentration increase of N-acylhomoserine lactone (AHL), which can diffuse in and out of the cell. Effective control of QS is expected to avoid virulence factor production in infectious pathogens, biofilm formation, and antibiotic production because various cell functions in gram-negative bacteria are controlled by AHL-mediated QS. In this research, we applied cyclodextrins (CDs) as artificial hosts for the AHL signal to reduce the AHL concentration in the culture broth below its threshold for QS activation. The AHL-receptor complex induced under the high AHL concentration activates transcription of the QS-target gene. Accordingly, artificial reduction of the AHL concentration is one of the effective strategies to inhibit the QS. A hydrophobic cavity of the CD can interact with the acyl-chain of the AHL due to hydrophobic interaction in aqueous media. We studied N-hexanoylhomoserine lactone (C6HSL)-mediated QS in Serratia marcescens; accumulation of C6HSL is responsible for regulation of the expression of pig cluster. Inhibitory effects of added CDs on QS were demonstrated by determination of prodigiosin amount inside cells after reaching stationary phase, because production of prodigiosin depends on the C6HSL-mediated QS. By adding approximately 6 wt% hydroxypropyl-β-CD (HP-β-CD) in Luria-Bertani (LB) medium prior to inoculation of S. maecescens AS-1, the intracellularly accumulated prodigiosin was drastically reduced to 7-10%, which was determined after the extraction of prodigiosin in acidified ethanol. The AHL retention ability of HP-β-CD was also demonstrated by Chromobacterium violacuem CV026 bioassay. The CV026 strain is an AHL-synthase defective mutant that activates QS solely by adding AHLs from outside of cells. A purple pigment violacein is induced by activation of the AHL-mediated QS. We demonstrated that the violacein production was effectively suppressed when the C6HSL standard solution was spotted on a LB agar plate dispersing CV026 cells and HP-β-CD. Physico-chemical analysis was performed to study the affinity between the immobilized CD and added C6HSL using a quartz crystal microbalance (QCM) sensor. The COOH-terminated self-assembled monolayer was prepared on a gold electrode of 27-MHz AT-cut quartz crystal. Mono(6-deoxy-6-N, N-diethylamino)-β-CD was immobilized on the electrode using water-soluble carbodiimide. The C6HSL interaction with the β-CD cavity was studied by injecting the C6HSL solution to a cup-type sensor cell filled with buffer solution. A decrement of resonant frequency (ΔFs) clearly showed the effective C6HSL complexation with immobilized β-CD and its stability constant for MBP-SpnR-C6HSL complex was on the order of 102 M-1. The CD has high potential for engineered control of QS because it is safe for human use.

Keywords: acylhomoserine lactone, cyclodextrin, intracellular signaling, quorum sensing

Procedia PDF Downloads 241

Authors: Anuj Kumar, Kummara M. Rao, Sung S. Han

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Bone tissue engineering has emerged as a potentially alternative method for localized bone defects or diseases, congenital deformation, and surgical reconstruction. The designing and the fabrication of the ideal scaffold is a great challenge, in restoring of the damaged bone tissues via cell attachment, proliferation, and differentiation under three-dimensional (3D) biological micro-/nano-environment. In this case, hydrogel system composed of high hydrophilic 3D polymeric-network that is able to mimic some of the functional physical and chemical properties of the extracellular matrix (ECM) and possibly may provide a suitable 3D micro-/nano-environment (i.e., resemblance of native bone tissues). Thus, this proposed hydrogel system is highly permeable and facilitates the transport of the nutrients and metabolites. However, the use of hydrogels in bone tissue engineering is limited because of their low mechanical properties (toughness and stiffness) that continue to posing challenges in designing and fabrication of tough and stiff hydrogels along with improved bioactive properties. For this purpose, in our lab, polyacrylamide-based hybrid hydrogels were synthesized by involving sodium alginate, cellulose nanocrystals and silica-based glass using one-step free-radical polymerization. The results showed good in vitro apatite-forming ability (biomineralization) and improved mechanical properties (under compression in the form of strength and stiffness in both wet and dry conditions), and in vitro osteoblastic (MC3T3-E1 cells) cytocompatibility. For in vitro cytocompatibility assessment, both qualitative (attachment and spreading of cells using FESEM) and quantitative (cell viability and proliferation using MTT assay) analyses were performed. The obtained hybrid hydrogels may potentially be used in bone tissue engineering applications after establishment of in vivo characterization.

Keywords: bone tissue engineering, cellulose nanocrystals, hydrogels, polyacrylamide, sodium alginate

Procedia PDF Downloads 152

Authors: Eugenia Vomvoridi-Ivanovic

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It is broadly assumed that participation in field experiences will help pre-service teachers (PSTs) bridge theory to practice. However, this is often not the case since PSTs who are placed in classrooms with large numbers of students from diverse linguistic, cultural, racial, and ethnic backgrounds (culturally diverse students (CDS)) usually observe ineffective mathematics teaching practices that are in contrast to those discussed in their teacher preparation program. Over the past decades, the educational research community has paid increasing attention to investigating out-of-school learning contexts and how participation in such contexts can contribute to the achievement of underrepresented groups in Science, Technology, Engineering, and mathematics (STEM) education and their expanded participation in STEM fields. In addition, several research studies have shown that students display different kinds of mathematical behaviors and discourse practices in out-of-school contexts than they do in the typical mathematics classroom since they draw from a variety of linguistic and cultural resources to negotiate meanings and participate in joint problem solving. However, almost no attention has been given to exploring these contexts as field experiences for pre-service mathematics teachers. The purpose of this study was to explore how participation in a community based after-school field experience promotes understanding of the content pedagogy concepts introduced in elementary mathematics methods courses, particularly as they apply to teaching mathematics to CDS. This study draws upon a situated, socio-cultural theory of teacher learning that centers on the concept of learning as situated social practice, which includes discourse, social interaction, and participation structures. Consistent with exploratory case study methodology, qualitative methods were employed to investigate how a cohort of twelve participating pre-service teacher's approach to pedagogy and their conversations around teaching and learning mathematics to CDS evolved through their participation in the after-school field experience, and how they connected the content discussed in their mathematics methods course with their interactions with the CDS in the after-school. Data were collected over a period of one academic year from the following sources: (a) audio recordings of the PSTs' interactions with the students during the after-school sessions, (b) PSTs' after-school field-notes, (c) audio-recordings of weekly methods course meetings, and (d) other document data (e.g., PST and student generated artifacts, PSTs' written course assignments). The findings of this study reveal that the PSTs benefitted greatly through their participation in the after-school field experience. Specifically, after-school participation promoted a deeper understanding of the content pedagogy concepts introduced in the mathematics methods course and gained a greater appreciation for how students learn mathematics with understanding. Further, even though many of PSTs' assumptions about the mathematical abilities of CDS were challenged and PSTs began to view CDSs' cultural and linguistic backgrounds as resources (rather than obstacles) for learning, some PSTs still held negative stereotypes about CDS and teaching and learning mathematics to CDS in particular. Insights gained through this study contribute to a better understanding of how informal mathematics learning contexts may provide a valuable context for pre-service teacher's learning to teach mathematics to CDS.

Keywords: after-school mathematics program, pre-service mathematical education of teachers, qualitative methods, situated socio-cultural theory, teaching culturally diverse students

Procedia PDF Downloads 131

Authors: Muhammad Isah Legbo

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The phytochemical screening and antimicrobial activity of Nauclea lotifolia fruit, leaf and stem-bark extracts at various concentration of (20.0,10.0, 5.0, and 2.5 mg/ml) were evaluated against some pathogenic microorganisms such as Escherichia coli, Klebsiella pneumoniae, Salmonella typhi, Staphylococcus aureus, Aspergillus niger and Candida albicans. The antimicrobial activity was assayed using agar well diffusion method. The result obtained show appreciable inhibitory effort of acetone, aqueous and methanolic extracts of Nauclea lotifolia. However, result obtained was less active compared to that of the control antibiotic (Ciprofloxacillin). The minimum inhibitory concentration (MIC) was determined using serial doubling dilution method and ranged from 5.0-10.0mg/ml, while the minimum bactericidal concentration (MBC) was determined by plating various dilution of extracts without turbidity and the result ranged from 5.0-7.5mg/ml. The phytochemical screening revealed the presence of alkaloid, anthraquinones, flavonoids, resin, steroid and saponin. The activities of the plant extract therefore justify their utilization in the treatment of various ailments associated with the test organism.

Keywords: Nauclea, lotifolia, antimicrobial, pathogens, saponin, extract

Procedia PDF Downloads 415

Authors: Meiyan Xing, Cenran Li, Liang Xiang

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Impacts of excess sludge vermicompost on the germination and early growth of plant were tested. The better effect of cow dung vermicompost (CV) on seed germination and seedling growth proved that cow dung was indeed the preferred additive in sludge vermicomposting as reported by plentiful researchers worldwide. The effects and the best amount of application of CV were further discussed. Results demonstrated that seed germination and seedling growth (seedlings number, plant height, stem diameter) were the best and heavy metal (Zn, Pb, Cr and As) contents of plant were the lowest when soil amended with CV by 15%. Additionally, CV fostered higher contents of chlorophyll a and chlorophyll b compared to the control when concentration ranged from 5 to 15%, thereafter a slight increase in chlorophyll content was observed form 15% to 25%. Thus, CV at the optimum proportion of 15% could serve as a feasible and satisfactory way of sludge agricultural utilization of sewage sludge. In summary, sewage sludge can be gainfully utilized in producing organic fertilizer via vermicomposting, thereby not only providing a means of sewage sludge treatment and disposal, but also stimulating the growth of plant and the ability to resist disease.

Keywords: cow dung vermicompost, seed germination, seedling growth, sludge utilization

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Authors: Ramin Ghasemi Shayan, Morteza Janebifam

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Tumors are regularly inhomogeneous. Provincial varieties in death, metabolic action, multiplication and body part are watched. There’s expanding proof that strong tumors may contain subpopulations of cells with various genotypes and phenotypes. These unmistakable populaces of malignancy cells can connect during a serious way and may contrast in affectability to medications. Most tumors show organic heterogeneity1–3 remembering heterogeneity for genomic subtypes, varieties inside the statement of development variables and genius, and hostile to angiogenic factors4–9 and varieties inside the tumoural microenvironment. These can present as contrasts between tumors in a few people. for instance, O6-methylguanine-DNA methyltransferase, a DNA fix compound, is hushed by methylation of the quality advertiser in half of glioblastoma (GBM), adding to chemosensitivity, and improved endurance. From the outset, there includes been specific enthusiasm inside the usage of dissemination weighted imaging (DWI) and dynamic complexity upgraded MRI (DCE-MRI). DWI sharpens MRI to water dispersion inside the extravascular extracellular space (EES) and is wiped out with the size and setup of the cell populace. Additionally, DCE-MRI utilizes dynamic obtaining of pictures during and after the infusion of intravenous complexity operator. Signal changes are additionally changed to outright grouping of differentiation permitting examination utilizing pharmacokinetic models. PET scan modality gives one of a kind natural particularity, permitting dynamic or static imaging of organic atoms marked with positron emanating isotopes (for example, 15O, 18F, 11C). The strategy is explained to a colossal radiation portion, which points of confinement rehashed estimations, particularly when utilized together with PC tomography (CT). At long last, it's of incredible enthusiasm to quantify territorial hemoglobin state, which could be joined with DCE-CT vascular physiology estimation to create significant experiences for understanding tumor hypoxia.

Keywords: heterogeneity, computerized tomography scan, magnetic resonance imaging, PET

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Authors: Hossein Khara, Mahmoud Sayyadborani, Mohammad Sayyadborani

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In the present study, we examined the effects of different dietary levels of ascorbic acid (vitamin C) and α-tocopherol (vitamin E) and their combinations on growth, survival and some haematological and immunological parameters of Caspian brown trout, Salmo trutta caspius juveniles. 15 experimental treatments and one control group with three replicates were considered for experiment. The experimental treatments were fish fed by experimental diets containing different levels of Vit C and E as follow: T1: Vit E (20 mg.kg diet -1) + Vit C (100 mg.kg diet -1), T2: Vit E (30 mg.kg diet -1) + Vit C (100 mg.kg diet -1), T3: Vit E (40 mg.kg diet -1) + Vit C (100 mg.kg diet -1), T4: Vit E (20 mg.kg diet -1) + Vit C (200 mg.kg diet -1), T5: Vit E (30 mg.kg diet -1) + Vit C (200 mg.kg diet -1), T6: Vit E (40 mg.kg diet -1) + Vit C (200 mg.kg diet -1), T7: Vit E (20 mg.kg diet -1) + Vit C (300 mg.kg diet -1), T8: Vit E (30 mg.kg diet -1) + Vit C (300 mg.kg diet -1), T9: Vit E (40 mg.kg diet -1) + Vit C (300 mg.kg diet -1), T10: Vit C (100 mg.kg diet -1), T11: Vit C (200 mg.kg diet -1), T12: Vit C (300 mg.kg diet -1), T13: Vit E (20 mg.kg diet -1), T14: Vit E (30 mg.kg diet -1) T15: Vit E (40 mg.kg diet -1). Also a non-vitamin supplemented was considered as control group. Growth parameters were measured monthly and serum parameters assayed at the end of the experiment. According to our results, Vit C and E improved survival and growth parameters including specific growth rate (SGR), weight gain percent (WG%) and biomass. The highest values of these parameters obtained in T8, T9 and T8 respectively. The lowest FCR obtained in T8. The haematological parameters including red blood cells (RBCs), white blood cells (WBCs), haematocrit (Hct) and haemoglobin (Hb) were higher in vitamin treated groups than control group with highest values in T8. In T13, WBC values were higher compared to other experimental groups. The immunological parameters including lysozyme activity, Immunoglobulin (IgM) and total immunoglobulin (TIg) were significantly higher in vitamin supplemented groups than in control group. In this regard the highest values of these parameters were found in T12. The lowest values of TIg and lysozyme activity were observed in control group and fish fed by only vitamin E i.e. T13, T14 and T15. In conclusion, our results show that Vit C and E in combination or only can improve growth, survival, haematological and immunological indices of Caspian brown trout.

Keywords: vitamins E, vitamins C, growth, survival, haematological parameters, immunological parameters

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Authors: Arun Kharate, Sarata Kumar Sahu, Susen Kumar Panda, Niranjan Sahoo, H. K. Panda

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In the present study, an attempt has been made for isolation and identification of feline panleukopaenia virus (FPLV) from wild felids of Nandankanan zoo, Odisha, India, along with prevalence study of FPLV. Fecal samples collected from wild felids (26 tigers, 22 lions, 5 leopards, 3 hyenas, 1 jaguar, 2 foxes and 1 wild cat) were subjected to hemagglutinnation test and fluorescent antibody test. In hemagglutinnation test 13 (50%) samples from tiger, 14 (63.63%) samples from lions, 1 (20%) sample from leopards, 1 (50%) from fox, 3 (100%) samples from hyenas and 1 (100%) sample from wild cat were positive. On fluorescent antibody test (FAT), 15 (57.69%) samples from tiger, 18 (81.81%) from lions, 2 (40%) from leopards, 1 (50%) from fox, 3 (100%) from hyenas and 1 (100%) from wild cat were positive. FPLV was isolated using MDBK cell line and preliminary characterization was done on the basis of characteristic cytopathic effect. The virus samples were quantified through titration in MDBK cells. Serological confirmation of FPLV isolates was carried out by HI test, micro-SNT and indirect-ELISA. Physico-chemical characters like pH and temperature resistance along molecular identification using specific FPLV primers was carried out. Seroprevalence study of 36 serum samples employing HI test, micro SNT and indirect-ELISA revealed prevalence of 38.8, 44.4 and 72.2% respectively. During study period an adult tigress and a tiger cub died suspected of feline panleukopenia. The necropsy findings in both animals showed hemorrhagic gastroenteritis. The cytological examination revealed presence of intranuclear inclusion bodies in the intestinal epithelial cells. Spleen, mesenteric lymph node and intestine were positive for feline panleukopenia by FAT. The investigation revealed that feline panleukopenia was prevalent in wild felines of Nandankanan zoo.

Keywords: Feline panleukopenia, fluorescent antibody test, hemagglutination test, indirect-ELISA, Nandankanan zoo

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Authors: H. El-Messiry, M. El-Zainy, D. Ghazy

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Diabetes is a metabolic disease that results in a variety of oral health complications. Green tea is a natural antioxidant proved to have powerful effects against diabetes. The aim of this study was to compare between the effect of insulin and green tea on the Parotid gland of streptozotocin induced diabetic Albino rats by using light and transmission electron microscopy. Forty male Albino rats were divided into control group and diabetic groups. The diabetic group received a single injection of 40 mg/kg of streptozotocin intra-peritoneal under anesthesia and was further subdivided into three subgroups: The diabetic untreated subgroup which was untreated for two weeks, the insulin treated subgroup which has received insulin subcutaneously in a daily dose of 5 IU/kg body weight/day for two weeks and a green tea treated subgroup received a daily dose of 1 ml/ 100 gm body weight intragastrically for two weeks. Rats were terminated and parotid glands were dissected and processed for light and transmission electron microscopic examination. Histological examination of the diabetic untreated subgroup revealed acinar cells with pyknotic and hyperchromatic nuclei with cytoplasmic vacuolations. Ultrastructurally, acinar cells showed nuclear pleomorphism, dilated rough endoplasmic reticulum and swollen mitochondria with damaged cristae. Inflammatory cell infiltration was detected both histologically and ultrastructurally. Ducts showed signs of degeneration with loss of their normal outline and stagnated secretion within the lumen. However, insulin and green tea treated subgroups showed minimal degenerative damage and were almost similar to the control with minimal changes. Treatment of the parotid gland of the streptozotocin induced diabetic rats with GT was closely comparable to the traditional insulin therapy in reducing signs of histological and ultrastructural damage.

Keywords: diabetes, green tea, insulin, parotid

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Authors: H. Akman, A. Topal

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This work was conducted in greenhouse conditions in order to investigate root biomass growth of two bread wheat, two durum wheat and two barley cultivars that were grown in irrigated and dry lands, respectively. This work was planned with four replications at a Completely Randomized Block Design in 2011-2012 growing season. In the study, root biomass growth was evaluated at stages of stem elongation, complete of anthesis and full grain maturity. Results showed that there were significant differences between cultivars grown at dry and irrigated lands in all growth stages in terms of root biomass (P < 0.01). According to research results, all of growth stages, dry typed-bread and durum wheats generally had higher root biomass than irrigated typed-cultivars, furthermore that dry typed-barley cultivar, had higher root biomass at GS 31 and GS 69, however lower at GS 92 than Larende. In all cultivars, root biomass increased between GS 31 and GS 69 so that dry typed-cultivars had more root biomass increase than irrigated typed-cultivars. Root biomass of bread wheat increased between GS 69 and GS 92, however root biomass of barley and durum wheat decreased.

Keywords: bread and durum wheat, barley, root biomass, different growth stage

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Authors: Manon Giraud, Francois-Damien Delapierre, Guenaelle Jasmin-Lebras, Cecile Feraudet-Tarisse, Stephanie Simon, Claude Fermon

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Early diagnosis or detection of harmful substances at low level is a growing field of high interest. The ideal test should be cheap, easy to use, quick, reliable, specific, and with very low detection limit. Combining the high specificity of antibodies-functionalized magnetic beads used to immune-capture biologic objects and the high sensitivity of a GMR-based sensors, it is possible to even detect these biologic objects one by one, such as a cancerous cell, a bacteria or a disease biomarker. The simplicity of the detection process makes its use possible even for untrained staff. Giant Magneto Resistance (GMR) is a recently discovered effect consisting in the electrical resistance modification of some conductive layers when exposed to a magnetic field. This effect allows the detection of very low variations of magnetic field (typically a few tens of nanoTesla). Magnetic nanobeads coated with antibodies targeting the analytes are mixed with a biological sample (blood, saliva) and incubated for 45 min. Then the mixture is injected in a very simple microfluidic chip and circulates above a GMR sensor that detects changes in the surrounding magnetic field. Magnetic particles do not create a field sufficient to be detected. Therefore, only the biological objects surrounded by several antibodies-functionalized magnetic beads (that have been captured by the complementary antigens) are detected when they move above the sensor. Proof of concept has been carried out on NS1 mouse cancerous cells diluted in PBS which have been bonded to magnetic 200nm particles. Signals were detected in cells-containing samples while none were recorded for negative controls. Binary response was hence assessed for this first biological model. The precise quantification of the analytes and its detection in highly diluted solution is the step now in progress.

Keywords: early diagnosis, giant magnetoresistance, lab-on-a-chip, submicron particle

Procedia PDF Downloads 249

Authors: Elisabetta Ceretti, Silvia Bonizzoni, Alberto Bonetti, Milena Villarini, Marco Verani, Maria Antonella De Donno, Sara Bonetta, Umberto Gelatti

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Introduction: Air pollution is a global problem. In 2013, the International Agency for Research on Cancer (IARC) classified air pollution and particulate matter as carcinogenic to human. The study of the health effects of air pollution in children is very important because they are a high-risk group in terms of the health effects of air pollution and early exposure during childhood can increase the risk of developing chronic diseases in adulthood. The MAPEC_LIFE (Monitoring Air Pollution Effects on Children for supporting public health policy) is a project founded by EU Life+ Programme which intends to evaluate the associations between air pollution and early biological effects in children and to propose a model for estimating the global risk of early biological effects due to air pollutants and other factors in children. Methods: The study was carried out on 6-8-year-old children living in five Italian towns in two different seasons. Two biomarkers of early biological effects, primary DNA damage detected with the comet assay and frequency of micronuclei, were investigated in buccal cells of children. Details of children diseases, socio-economic status, exposures to other pollutants and life-style were collected using a questionnaire administered to children’s parents. Child exposure to urban air pollution was assessed by analysing PM0.5 samples collected in the school areas for PAHs and nitro-PAHs concentration, lung toxicity and in vitro genotoxicity on bacterial and human cells. Data on the chemical features of the urban air during the study period were obtained from the Regional Agency for Environmental Protection. The project created also the opportunity to approach the issue of air pollution with the children, trying to raise their awareness on air quality, its health effects and some healthy behaviors by means of an educational intervention in the schools. Results: 1315 children were recruited for the study and participate in the first sampling campaign in the five towns. The second campaign, on the same children, is still ongoing. The preliminary results of the tests on buccal mucosa cells of children will be presented during the conference as well as the preliminary data about the chemical composition and the toxicity and genotoxicity features of PM0.5 samples. The educational package was tested on 250 children of the primary school and showed to be very useful, improving children knowledge about air pollution and its effects and stimulating their interest. Conclusions: The associations between levels of air pollutants, air mutagenicity and biomarkers of early effects will be investigated. A tentative model to calculate the global absolute risk of having early biological effects for air pollution and other variables together will be proposed and may be useful to support policy-making and community interventions to protect children from possible health effects of air pollutants.

Keywords: air pollution exposure, biomarkers of early effects, children, public health policy

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Authors: Hediyeh Talebi, Shokoofeh Ghiam, Changiz Eslahchi

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Traditionally, Alzheimer’s disease research has focused on genes with significant fold changes, potentially neglecting subtle but biologically important alterations. Our study introduces an integrative approach that highlights genes crucial to underlying biological processes, regardless of their fold change magnitude. Alzheimer's Single-cell RNA-seq data related to the peripheral blood mononuclear cells (PBMC) was extracted from the Gene Expression Omnibus (GEO). After quality control, normalization, scaling, batch effect correction, and clustering, differentially expressed genes (DEGs) were identified with adjusted p-values less than 0.05. These DEGs were categorized based on cell-type, resulting in four datasets, each corresponding to a distinct cell type. To distinguish between cells from healthy individuals and those with Alzheimer's, an adversarial autoencoder with a classifier was employed. This allowed for the separation of healthy and diseased samples. To identify the most influential genes in this classification, the weight matrices in the network, which includes the encoder and classifier components, were multiplied, and focused on the top 20 genes. The analysis revealed that while some of these genes exhibit a high fold change, others do not. These genes, which may be overlooked by previous methods due to their low fold change, were shown to be significant in our study. The findings highlight the critical role of genes with subtle alterations in diagnosing Alzheimer's disease, a facet frequently overlooked by conventional methods. These genes demonstrate remarkable discriminatory power, underscoring the need to integrate biological relevance with statistical measures in gene prioritization. This integrative approach enhances our understanding of the molecular mechanisms in Alzheimer’s disease and provides a promising direction for identifying potential therapeutic targets.

Keywords: alzheimer's disease, single-cell RNA-seq, neural networks, blood biomarkers

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Authors: Kuladip Jana

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Benzo(a)pyrene [B(a)P] is an environmental toxicant present mostly in cigarette smoke and car exhaust, is an aryl hydrocarbon receptor (AhR) ligand that exerts its toxic effects on both male and female reproductive systems. In this study, the effect of B(a)P at different doses (0.1, 0.25, 0.5, 1 and 5 mg /kg body weight) was studied on male reproductive system of rat. A significant decrease in cauda epididymal sperm count and motility along with the presence of sperm head abnormalities and altered epididymal and testicular histology were documented following B(a)P treatment. B(a)P treatment resulted apoptotic sperm cells as observed by TUNEL and Annexin V-PI assay with increased ROS, altered sperm mitochondrial membrane potential (ΔΨm) with a simultaneous decrease in the activity of antioxidant enzymes and GSH status. TUNEL positive apoptotic cells also observed in testis as well as isolated germ and Leydig cells following B(a)P exposure. Western Blot analysis revealed the activation of p38MAPK, cytosolic translocation of cytochrome-c, up-regulation of Bax and inducible nitric oxide synthase (iNOS) with cleavage of PARP and down-regulation of BCl2 in testis upon B(a)P treatment. The protein and mRNA levels of testicular key steroidogenesis regulatory proteins like StAR, cytochrome P450 IIA1 (CYPIIA1), 3β HSD, 17β HSD showed a significant decrease in a dose dependent manner while an increase in the expression of cytochrome P450 1A1 (CYP1A1), Aryl hydrocarbon Receptor (AhR), active caspase- 9 and caspase- 3 following B(a)P exposure. We conclude that exposure of benzo(a)pyrene caused testicular gamatogenic and steroidogenic disorders by induction of oxidative stress, inhibition of StAR and other steroidogenic enzymes along with activation of p38MAPK and initiated caspase-3 mediated germ and Leydig cell apoptosis.The possible protective role of naturally occurring phytochemicals against B(a)P induced testicular toxicity needs immediate consideration. Curcumin and resveratrol separately were found to protect against B(a)P induced germ cell apoptosis, and their combinatorial effect was more significant. Our present study in isolated testicular germ cell population from adult male Wistar rats, highlighted their synergistic protective effect against B(a)P induced germ cell apoptosis. Curcumin-resveratrol co-treatment decreased the expression of pro-apoptotic proteins like cleaved caspase 3,8,9, cleaved PARP, Apaf1, FasL, tBid. Curcumin-resveratrol co-treatment decreased Bax/Bcl2 ratio, mitochondria to cytosolic translocation of cytochrome c and activated the survival protein Akt. Curcumin-resveratrol decreased the expression of p53 dependent apoptotic genes like Fas, FasL, Bax, Bcl2, Apaf1.Curcumin-resveratrol co-treatment thus prevented B(a)P induced germ cell apoptosis. B(a)P induced testicular ROS generation and oxidative stress were significantly ameliorated with curcumin and resveratrol. Curcumin-resveratrol co-treatment prevented B(a)P induced nuclear translocation of AhR and CYP1A1 production. The combinatorial treatment significantly inhibited B(a)P induced ERK 1/2, p38 MAPK and JNK 1/2 activation. B(a)P treatment increased the expression of p53 and its phosphorylation (p53 ser 15). Curcumin-resveratrol co-treatment significantly decreased p53 level and its phosphorylation (p53 ser 15). The study concludes that curcumin-resveratrol synergistically modulated MAPKs and p53, prevented oxidative stress, regulated the expression of pro and anti-apoptotic proteins as well as the proteins involved in B(a)P metabolism thus protected germ cells from B(a)P induced apoptosis.

Keywords: benzo(a)pyrene, germ cell, apoptosis, oxidative stress, resveratrol, curcumin

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Authors: M. P. Dandekar, A. P. Bharne, P. T. Borkar, D. M. Kokare, N. K. Subhedar

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Ethanol ingestion by the mother ensue adverse consequences for her offspring. Herein, we examine the behavioral phenotype and neural substrate of the offspring of the mother on ethanol. Female rats were fed with ethanol-containing liquid diet from 8 days prior of conception and continued till 25 days post-parturition to coincide with weaning. Behavioral changes associated with anxiety, depression and learning and memory were assessed in the offspring, after they attained adulthood (day 85), using elevated plus maze (EPM), forced swim (FST) and novel object recognition tests (NORT), respectively. The offspring of the alcoholic mother, compared to those of the pair-fed mother, spent significantly more time in closed arms of EPM and showed more immobility time in FST. Offspring at the age of 25 and 85 days failed to discriminate between novel versus familiar object in NORT, thus reflecting anxiogenic, depressive and amnesic phenotypes. Neuropeptide cocaine- and amphetamine-regulated transcript peptide (CART) is known to be involved in central effects of ethanol and hence selected for the current study. Twenty-five days old pups of the alcoholic mother showed significant augmentation in CART-immunoreactivity in the cells of Edinger-Westphal (EW) nucleus and lateral hypothalamus. However, a significant decrease in CART-immunoreactivity was seen in nucleus accumbens shell (AcbSh), lateral part of bed nucleus of the stria terminalis (BNSTl), locus coeruleus (LC), hippocampus (CA1, CA2 and CA3), and arcuate nucleus (ARC) of the pups and/or adults offspring. While no change in the CART-immunoreactive fibers of AcbSh and BNSTl, CA2 and CA3 was noticed in the 25 days old pups, the CART-immunoreactive cells in EW and paraventricular nucleus (PVN), and fibers in the central nucleus of amygdala of 85 days old offspring remained unaffected. We suggest that the endogenous CART system in these discrete areas, among other factors, may be a causal to the abnormalities in the next generation of an alcoholic mother.

Keywords: anxiety, depression, CART, ethanol, immunocytochemistry

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