Search results for: red blood cell
5669 Blood Clot Emulsification via Ultrasonic Thrombolysis Device
Authors: Sun Tao, Lou Liang, Tan Xing Haw Marvin, Gu Yuandong Alex
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Patients with blood clots in their brains can experience problems with their vision or speech, seizures and general weakness. To treat blood clots, clinicians presently have two options. The first involves drug therapy to thin the blood and thus reduce the clot. The second choice is to invasively remove the clot using a plastic tube called a catheter. Both approaches carry a high risk of bleeding, and invasive procedures, such as catheter intervention, can also damage the blood vessel wall and cause infection. Ultrasonic treatment as a potential alternative therapy to break down clots is attracting growing interests due to the reduced adverse effects. To demonstrate the concept, in this investigation a microfabricated ultrasonic device was electrically packaged with printed circuit board to treat healthy human blood. The red blood cells could be broken down after 3-hour ultrasonic treatment.Keywords: microfabrication, blood clot, ultrasonic thrombolysis device, ultrasonic device
Procedia PDF Downloads 4515668 Potential Therapeutic Effect of Obestatin in Oral Mucositis
Authors: Agnieszka Stempniewicz, Piotr Ceranowicz, Wojciech Macyk, Jakub Cieszkowski, Beata Kuśnierz-Cabała, Katarzyna Gałązka, Zygmunt Warzecha
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Objectives: There are numerous strategies for the prevention or treatment of oral mucositis. However, their effectiveness is limited and does not correspond to expectations. Recent studies have shown that obestatin exhibits a protective effect and accelerates the healing of gastrointestinal mucosa. The aim of the present study was to examine the influence of obestatin administration on oral ulcers in rats. Methods: lingual ulcers were induced by the use of acetic acid. Rats were treated twice a day intraperitoneally with saline or obestatin(4, 8, or 16 nmol/kg/dose) for five days. The study determined: lingual mucosa morphology, cell proliferation, mucosal blood flow, and mucosal pro-inflammatory interleukin-1β level(IL-1β). Results: In animals without induction of oral ulcers, treatment with obestatin was without any effect. Obestatin administration in rats with lingual ulcers increased the healing rate of these ulcers. Obestatin given at the dose of 8 or 16 nmol/kg/dose caused the strongest and similar therapeutic effect. This result was associated with a significant increase in blood flow and cell proliferation in gingival mucosa, as well as a significant decrease in IL-1β level. Conclusions: Obestatin accelerates the healing of lingual ulcers in rats. This therapeutic effect is well-correlated with an increase in blood flow and cell proliferation in oral mucosa, as well as a decrease in pro-inflammatory IL-1β levels. Obestatin is a potentially useful candidate for the prevention and treatment of oral mucositis. Acknowledgment: Agnieszka Stempniewicz acknowledges the support of InterDokMed project no. POWR.03.02.00- 00-I013/16.Keywords: oral mucositis, ulcers, obestatin, lingual mucosa
Procedia PDF Downloads 735667 The Evaluation of Complete Blood Cell Count-Based Inflammatory Markers in Pediatric Obesity and Metabolic Syndrome
Authors: Mustafa M. Donma, Orkide Donma
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Obesity is defined as a severe chronic disease characterized by a low-grade inflammatory state. Therefore, inflammatory markers gained utmost importance during the evaluation of obesity and metabolic syndrome (MetS), a disease characterized by central obesity, elevated blood pressure, increased fasting blood glucose and elevated triglycerides or reduced high density lipoprotein cholesterol (HDL-C) values. Some inflammatory markers based upon complete blood cell count (CBC) are available. In this study, it was questioned which inflammatory marker was the best to evaluate the differences between various obesity groups. 514 pediatric individuals were recruited. 132 children with MetS, 155 morbid obese (MO), 90 obese (OB), 38 overweight (OW) and 99 children with normal BMI (N-BMI) were included into the scope of this study. Obesity groups were constituted using age- and sex-dependent body mass index (BMI) percentiles tabulated by World Health Organization. MetS components were determined to be able to specify children with MetS. CBC were determined using automated hematology analyzer. HDL-C analysis was performed. Using CBC parameters and HDL-C values, ratio markers of inflammation, which cover neutrophil-to-lymphocyte ratio (NLR), derived neutrophil-to-lymphocyte ratio (dNLR), platelet-to-lymphocyte ratio (PLR), lymphocyte-to-monocyte ratio (LMR), monocyte-to-HDL-C ratio (MHR) were calculated. Statistical analyses were performed. The statistical significance degree was considered as p < 0.05. There was no statistically significant difference among the groups in terms of platelet count, neutrophil count, lymphocyte count, monocyte count, and NLR. PLR differed significantly between OW and N-BMI as well as MetS. Monocyte-to HDL-C value exhibited statistical significance between MetS and N-BMI, OB, and MO groups. HDL-C value differed between MetS and N-BMI, OW, OB, MO groups. MHR was the ratio, which exhibits the best performance among the other CBC-based inflammatory markers. On the other hand, when MHR was compared to HDL-C only, it was suggested that HDL-C has given much more valuable information. Therefore, this parameter still keeps its value from the diagnostic point of view. Our results suggest that MHR can be an inflammatory marker during the evaluation of pediatric MetS, but the predictive value of this parameter was not superior to HDL-C during the evaluation of obesity.Keywords: children, complete blood cell count, high density lipoprotein cholesterol, metabolic syndrome, obesity
Procedia PDF Downloads 1305666 Global Analysis of HIV Virus Models with Cell-to-Cell
Authors: Hossein Pourbashash
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Recent experimental studies have shown that HIV can be transmitted directly from cell to cell when structures called virological synapses form during interactions between T cells. In this article, we describe a new within-host model of HIV infection that incorporates two mechanisms: infection by free virions and the direct cell-to-cell transmission. We conduct the local and global stability analysis of the model. We show that if the basic reproduction number R0 1, the virus is cleared and the disease dies out; if R0 > 1, the virus persists in the host. We also prove that the unique positive equilibrium attracts all positive solutions under additional assumptions on the parameters.Keywords: HIV virus model, cell-to-cell transmission, global stability, Lyapunov function, second compound matrices
Procedia PDF Downloads 5185665 Integrated Mathematical Modeling and Advance Visualization of Magnetic Nanoparticle for Drug Delivery, Drug Release and Effects to Cancer Cell Treatment
Authors: Norma Binti Alias, Che Rahim Che The, Norfarizan Mohd Said, Sakinah Abdul Hanan, Akhtar Ali
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This paper discusses on the transportation of magnetic drug targeting through blood within vessels, tissues and cells. There are three integrated mathematical models to be discussed and analyze the concentration of drug and blood flow through magnetic nanoparticles. The cell therapy brought advancement in the field of nanotechnology to fight against the tumors. The systematic therapeutic effect of Single Cells can reduce the growth of cancer tissue. The process of this nanoscale phenomena system is able to measure and to model, by identifying some parameters and applying fundamental principles of mathematical modeling and simulation. The mathematical modeling of single cell growth depends on three types of cell densities such as proliferative, quiescent and necrotic cells. The aim of this paper is to enhance the simulation of three types of models. The first model represents the transport of drugs by coupled partial differential equations (PDEs) with 3D parabolic type in a cylindrical coordinate system. This model is integrated by Non-Newtonian flow equations, leading to blood liquid flow as the medium for transportation system and the magnetic force on the magnetic nanoparticles. The interaction between the magnetic force on drug with magnetic properties produces induced currents and the applied magnetic field yields forces with tend to move slowly the movement of blood and bring the drug to the cancer cells. The devices of nanoscale allow the drug to discharge the blood vessels and even spread out through the tissue and access to the cancer cells. The second model is the transport of drug nanoparticles from the vascular system to a single cell. The treatment of the vascular system encounters some parameter identification such as magnetic nanoparticle targeted delivery, blood flow, momentum transport, density and viscosity for drug and blood medium, intensity of magnetic fields and the radius of the capillary. Based on two discretization techniques, finite difference method (FDM) and finite element method (FEM), the set of integrated models are transformed into a series of grid points to get a large system of equations. The third model is a single cell density model involving the three sets of first order PDEs equations for proliferating, quiescent and necrotic cells change over time and space in Cartesian coordinate which regulates under different rates of nutrients consumptions. The model presents the proliferative and quiescent cell growth depends on some parameter changes and the necrotic cells emerged as the tumor core. Some numerical schemes for solving the system of equations are compared and analyzed. Simulation and computation of the discretized model are supported by Matlab and C programming languages on a single processing unit. Some numerical results and analysis of the algorithms are presented in terms of informative presentation of tables, multiple graph and multidimensional visualization. As a conclusion, the integrated of three types mathematical modeling and the comparison of numerical performance indicates that the superior tool and analysis for solving the complete set of magnetic drug delivery system which give significant effects on the growth of the targeted cancer cell.Keywords: mathematical modeling, visualization, PDE models, magnetic nanoparticle drug delivery model, drug release model, single cell effects, avascular tumor growth, numerical analysis
Procedia PDF Downloads 4285664 The Admitting Hemogram as a Predictor for Severity and in-Hospital Mortality in Acute Pancreatitis
Authors: Florge Francis A. Sy
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Acute pancreatitis (AP) is an inflammatory condition of the pancreas with local and systemic complications. Severe acute pancreatitis (SAP) has a higher mortality rate. Laboratory parameters like the neutrophil-to-lymphocyte ratio (NLR), red cell distribution width (RDW), and mean platelet volume (MPV) have been associated with SAP but with conflicting results. This study aims to determine the predictive value of these parameters on the severity and in-hospital mortality of AP. This retrospective, cross-sectional study was done in a private hospital in Cebu City, Philippines. One-hundred five patients were classified according to severity based on the modified Marshall scoring. The admitting hemogram, including the NLR, RDW, and MPV, was obtained from the complete blood count (CBC). Cut-off values for severity and in-hospital mortality were derived from the ROC. Association between NLR, RDW, and MPV with SAP and mortality were determined with a p-value of < 0.05 considered significant. The mean age for AP was 47.6 years, with 50.5% being male. Most had an unknown cause (49.5%), followed by a biliary cause (37.1%). Of the 105 patients, 23 patients had SAP, and 4 died. Older age, longer in-hospital duration, congestive heart failure, elevated creatinine, urea nitrogen, and white blood cell count were seen in SAP. The NLR was associated with in-hospital mortality using a cut-off of > 10.6 (OR 1.133, 95% CI, p-value 0.003) with 100% sensitivity, 70.3% specificity, 11.76% PPV and 100% NPV (AUC 0.855). The NLR was not associated with SAP. The RDW and MPV were not associated with SAP and mortality. The admitting NLR is, therefore, an easily accessible parameter that can predict in-hospital mortality in acute pancreatitis. Although the present study did not show an association of NLR with SAP nor RDW and MPV with both SAP and mortality, further studies are suggested to establish their clinical value.Keywords: acute pancreatitis, mean platelet volume, neutrophil-lymphocyte ratio, red cell distribution width
Procedia PDF Downloads 1255663 IL-21 Production by CD4+ Effector T Cells and Frequency of Circulating Follicular Helper T Cells Are Increased in Type 1 Diabetes Patients
Authors: Ferreira RC, Simons HZ, Thompson WS, Cutler AJ, Dopico XC, Smyth DJ, Mashar M, Schuilenburg H, Walker NM, Dunger DB, Wallace C, Todd JA, Wicker LS, Pekalski ML
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Type 1 diabetes is caused by autoimmune destruction of insulin-secreting beta cells in the pancreas. T cells are known to play an important role in this immune-mediated destruction; however, there is no general consensus regarding alterations in cytokine production or T cell subsets in peripheral blood of patients with type 1 diabetes. Using polychromatic flow cytometry of peripheral blood mononuclear cells (PBMCs), we assessed production of the proinflammatory cytokines IL-21, IFN-γ and IL-17 by memory CD4 T effector (Teff) cells in 69 patients with type 1 diabetes and 61 healthy donors. We found a 21.9% (95% CI 5.8, 40.2; p = 3.9 × 10(-3)) higher frequency of IL-21(+) CD45RA(-) memory CD4(+) Teffs in patients with type 1 diabetes (geometric mean 5.92% [95% CI 5.44, 6.44]) compared with healthy donors (geometric mean 4.88% [95% CI 4.33, 5.50]). In a separate cohort of 30 patients with type 1 diabetes and 32 healthy donors, we assessed the frequency of circulating T follicular helper (Tfh) cells in whole blood. Consistent with the increased production of IL-21, we also found a 14.9% increase in circulating Tfh cells in the patients with type 1 diabetes (95% CI 2.9, 26.9; p = 0.016). Analysis of IL-21 production by PBMCs from a subset of 46 of the 62 donors immunophenotyped for Tfh showed that frequency of Tfh cells was associated with the frequency of IL-21+ cells (r2 = 0.174, p = 0.004). These results indicate that increased IL-21 production is likely to be an aetiological factor in the pathogenesis of type 1 diabetes that could be considered as a potential therapeutic target.Keywords: T follicular helper cell, IL-21, IL-17, type 1 diabetes
Procedia PDF Downloads 3815662 Expression of ULK-1 mRNA in Human Peripheral Blood Mononuclear Cells from Patients with Alzheimer's Disease
Authors: Ali Bayram, Remzi Yiğiter
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Objective: Alzheimer's disease (AD), the most common cause of dementia, is a progressive neurodegenerative disease. At present, diagnosis of AD is rather late in the disease. Therefore, we attempted to find peripheral biomarkers for the early diagnosis of AD. Herein, we conducted a study to investigate the unc-51 like autophagy activating kinase-1 (ULK1) mRNA expression levels in human peripheral blood mononuclear cells from patients with Alzheimer's disease. Method: To determine whether ULK1 gene expression are altered in AD patients, we measured their gene expression in human peripheral blood cell in 50 patients with AD and 50 age and gender matched healthy controls by quantitative real-time PCR technique. Results: We found that both ULK1 gene expression in peripheral blood cell were significantly decreased in patients with AD as compared with controls (p <0.05). Lower levels of ULK1 gene expression were significantly associated with the increased risk for AD. Conclusions: Serine/threonine-protein kinase involved in autophagy in response to starvation. Acts upstream of phosphatidylinositol 3-kinase PIK3C3 to regulate the formation of autophagophores, the precursors of autophagosomes. Part of regulatory feedback loops in autophagy: acts both as a downstream effector and negative regulator of mammalian target of rapamycin complex 1 (mTORC1) via interaction with RPTOR. Activated via phosphorylation by AMPK and also acts as a regulator of AMPK by mediating phosphorylation of AMPK subunits PRKAA1, PRKAB2, and PRKAG1, leading to negatively regulate AMPK activity. May phosphorylate ATG13/KIAA0652 and RPTOR; however such data need additional evidences. Plays a role early in neuronal differentiation and is required for granule cell axon formation. Alzheimer is the most common neurodegenerative disease. Our results provide useful information that the ULK1 gene expression is decreased in the neurodegeneration and AD patients with, indicating their possible systemic involvement in AD.Keywords: Alzheimer’s sisease, ULK1, mRNA expression, RT-PCR
Procedia PDF Downloads 3985661 Feasibility of Leukemia Cancer Treatment (K562) by Atmospheric Pressure Plasma Jet
Authors: Mashayekh Amir Shahriar, Akhlaghi Morteza, Rajaee Hajar, Khani Mohammad Reza, Shokri Babak
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A new and novel approach in medicine is the use of cold plasma for various applications such as sterilization blood coagulation and cancer cell treatment. In this paper a pin-to-hole plasma jet suitable for biological applications is investigated, characterized and the possibility and feasibility of cancer cell treatment is evaluated. The characterization includes power consumption via Lissajous method, thermal behavior of plasma using Infra-red camera as a novel method, Optical Emission Spectroscopy (OES) to determine the species that are generated. Treatment of leukemia cancer cells is also implemented and MTT assay is used to evaluate viability.Keywords: Atmospheric Pressure Plasma Jet (APPJ), Plasma Medicine, Cancer cell treatment, leukemia, Optical Emission
Procedia PDF Downloads 6615660 Numerical Analysis of Core-Annular Blood Flow in Microvessels at Low Reynolds Numbers
Authors: L. Achab, F. Iachachene
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In microvessels, red blood cells (RBCs) exhibit a tendency to migrate towards the vessel center, establishing a core-annular flow pattern. The core region, marked by a high concentration of RBCs, is governed by significantly non-Newtonian viscosity. Conversely, the annular layer, composed of cell-free plasma, is characterized by Newtonian low viscosity. This property enables the plasma layer to act as a lubricant for the vessel walls, efficiently reducing resistance to the movement of blood cells. In this study, we investigate the factors influencing blood flow in microvessels and the thickness of the annular plasma layer using a non-miscible fluids approach in a 2D axisymmetric geometry. The governing equations of an incompressible unsteady flow are solved numerically through the Volume of Fluid (VOF) method to track the interface between the two immiscible fluids. To model blood viscosity in the core region, we adopt the Quemada constitutive law which is accurately captures the shear-thinning blood rheology over a wide range of shear rates. Our results are then compared to an established theoretical approach under identical flow conditions, particularly concerning the radial velocity profile and the thickness of the annular plasma layer. The simulation findings for low Reynolds numbers, demonstrate a notable agreement with the theoretical solution, emphasizing the pivotal role of blood’s rheological properties in the core region in determining the thickness of the annular plasma layer.Keywords: core-annular flows, microvessels, Quemada model, plasma layer thickness, volume of fluid method
Procedia PDF Downloads 575659 Tumour-Associated Tissue Eosinophilia as a Prognosticator in Oral Squamous Cell Carcinoma
Authors: Karen Boaz, C. R. Charan
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Background: The infiltration of tumour stroma by eosinophils, Tumor-Associated Tissue Eosinophilia (TATE), is known to modulate the progression of Oral Squamous Cell Carcinoma (OSCC). Eosinophils have direct tumoricidal activity by release of cytotoxic proteins and indirectly they enhance permeability into tumor cells enabling penetration of tumoricidal cytokines. Also, eosinophils may promote tumor angiogenesis by production of several angiogenic factors. Identification of eosinophils in the inflammatory stroma has been proven to be an important prognosticator in cancers of mouth, oesophagus, larynx, pharynx, breast, lung, and intestine. Therefore, the study aimed to correlate TATE with clinical and histopathological variables, and blood eosinophil count to assess the role of TATE as a prognosticator in Oral Squamous Cell Carcinoma (OSCC). Methods: Seventy two biopsy-proven cases of OSCC formed the study cohort. Blood eosinophil counts and TNM stage were obtained from the medical records. Tissue sections (5µm thick) were stained with Haematoxylin and Eosin. The eosinophils were quantified at invasive tumour front (ITF) in 10HPF (40x magnification) with an ocular grid. Bryne’s grading of ITF was also performed. A subset of thirty cases was also assessed for association of TATE with recurrence, involvement of lymph nodes and surgical margins. Results: 1) No statistically significant correlation was found between TATE and TNM stage, blood eosinophil counts and most parameters of Bryne’s grading system. 2) Statistically significant relation of intense degree of TATE was associated with the absence of distant metastasis, increased lympho-plasmacytic response and increased survival (diseasefree and overall) of OSCC patients. 3) In the subset of 30 cases, tissue eosinophil counts were higher in cases with lymph node involvement, decreased survival, without margin involvement and in cases that did not recur. Conclusion: While the role of eosinophils in mediating immune responses seems ambiguous as eosinophils support cell-mediated tumour immunity in early stages while inhibiting the same in advanced stages, TATE may be used as a surrogate marker for determination of prognosis in oral squamous cell carcinoma.Keywords: tumour-associated tissue eosinophilia, oral squamous cell carcinoma, prognosticator, tumoral immunity
Procedia PDF Downloads 2505658 The Influence of Bacteriocins Producing Lactic Acid Bacteria Multiplied in an Alternative Substrate on Calves Blood Parameters
Authors: E. Bartkiene, V. Krungleviciute, J. Kucinskiene, R. Antanaitis, A. Kucinskas
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In calves less than 10-day-old, infection commonly cause severe diarrhoea and high mortality. To prevention of calves diseases a common practice is to treat calves with prophylactic antibiotics, in this case the use of lactic acid bacteria (LAB) is promising. Often LAB strains are incubated in comercial de Man-Rogosa-Sharpe (MRS) medium, the culture are centrifuged, the cells are washing with sterile water, and this suspension is used as a starter culture for animal health care. Juice of potatoe tubers is industrial wastes, wich may constitute a source of digestible nutrients for microorganisms. In our study the ability of LAB to utilize potatoe tubers juice in cell synthesis without external nutrient supplement was investigated, and the influence of multiplied LAB on calves blood parameters was evaluated. Calves were selected based on the analogy principle (treatment group (n=6), control group (n=8)). For the treatment group 14 days was given a 50 ml of fermented potatoe tubers juice containing 9.6 log10 cfu/ml of LAB. Blood parameters (gas and biochemical) were assessed by use of an auto-analyzers (Hitachi 705 and EPOC). Before the experiment, blood pH of treatment group calves was 7.33, control – 7.36, whereas, after 14 days, 7.28 and 7.36, respectively. Calves blood pH in the treatment group remained stable over the all experiment period. Concentration of PCO2 in control calves group blood increased from 63.95 to 70.93, whereas, in the treatment group decreased from 63.08 to 60.71. Concentration of lactate in the treatment group decreased from 3.20 mmol/l to 2.64 mmol/l, whereas, in control - increased from 3.95 mmol/l to 4.29 mmol/l. Concentration of AST in the control calves group increased from 50.18 IU/L to 58.9 IU/L, whereas, in treatment group decreased from 49.82 IU/L to 33.1 IU/L. We conclude that the 50 ml of fermented potatoe tubers juice containing 9.6 log10 cfu/ml of LAB per day, by using 14 days, reduced risk of developing acidosis (stabilizes blood pH (p < 0.05)), reduces lactates and PCO2 concentration (p < 0.05) and risk of liver lesions (reduces AST concentration (p < 0.005)) in blood of calves.Keywords: alternative substrate, blood parameters, calves, lactic acid bacteria
Procedia PDF Downloads 3155657 Correlation of Hematological Indices with Fasting Blood Glucose Level and Anthropometric Measurements in Geriatric Diabetes Mellitus Subjects in Lagos State University Teaching Hospital, Ikeja, Lagos, Nigeria
Authors: Dada. O.Akinola, Uche. I. Ebele, Bamiro .A.Rafatu, Akinbami A. Akinsegun, Dada O. Adeyemi, Adeyemi. O. Ibukun, Okunowo O.Bolanle, Abdulateef O. Kareem, Ibrahim.N. Ismaila, Dosu Rihanat
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Background: Hyperglycaemia alters qualitatively and quantitatively all the full blood count parameters. The alterations among other factors are responsible for the macrovascular and microvascular complications associated with diabetes mellitus (DM). This study is aimed at correlating haematological parameters in DM subjects with their fasting blood glucose (FBG) and anthropometric parameters. Materials and Methods: This was a cross-sectional study of participants attending DM clinic of Lagos State University Teaching Hospital (LASUTH), Ikeja. The study recruited one hundred and two (102) DM subjects and one hundred (100) non-DM controls. Venous blood samples were collected for full blood count (FBC) assay while FBG was done, structured questionnaires were administered, and anthropometric measurements of all participants were done. Data were analyzed with Statistical Package for Social Science (SPSS) version 23. P was set at ≤0.05. Results: The mean age of DM patients was 64.32± 11.31 years. Using a haemoglobin concentration cut-off of 11g/dl, 39.2%, and 13% DM and control participants respectively had values lower than 11g/dl. A total of 22.5% and 3% of DM and controls respectively gave a history of previous blood transfusion.White blood cells count and platelet count means were (6.12±1.60 and 5.30±7.52,p=0.59) and (213.31±73.58 and 228.91±73.21,p = 0.26) *109/L in DM subjects and controls respectively. FBG and all the anthropometric data in DM subjects were significantly higher than in controls. Conclusions: The prevalence of anaemia in DM subjects was three times higher than in controls. The white blood cell count was higher but not statistically significant in DM compared with controls. But platelet count was higher but not statistically significant in controls compared with DM subjects.Keywords: haematological profile, diabetes mellitus, anthropometric data, fasting blood glucose
Procedia PDF Downloads 855656 Development of Monitoring Blood Bank Center Based PIC Microcontroller Using CAN Communication
Authors: Kaiwan S. Ismael, Ergun Ercelebi, Majeed Nader
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This paper describes the design and implementation of a hardware setup for online monitoring of 24 refrigerators inside blood bank center using the microcontroller and CAN bus for communications between each node. Due to the security of locations in the blood bank hall and difficulty of monitoring of each refrigerator separately, this work proposes a solution to monitor all the blood bank refrigerators in one location. CAN-bus system is used because it has many applications and advantages, especially for this system due to easy in use, low cost, providing a reduction in wiring, fast to repair and easily expanding the project without a problem.Keywords: control area network (CAN), monitoring blood bank center, PIC microcontroller, MPLAB IDE
Procedia PDF Downloads 4865655 Single-Cell Visualization with Minimum Volume Embedding
Authors: Zhenqiu Liu
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Visualizing the heterogeneity within cell-populations for single-cell RNA-seq data is crucial for studying the functional diversity of a cell. However, because of the high level of noises, outlier, and dropouts, it is very challenging to measure the cell-to-cell similarity (distance), visualize and cluster the data in a low-dimension. Minimum volume embedding (MVE) projects the data into a lower-dimensional space and is a promising tool for data visualization. However, it is computationally inefficient to solve a semi-definite programming (SDP) when the sample size is large. Therefore, it is not applicable to single-cell RNA-seq data with thousands of samples. In this paper, we develop an efficient algorithm with an accelerated proximal gradient method and visualize the single-cell RNA-seq data efficiently. We demonstrate that the proposed approach separates known subpopulations more accurately in single-cell data sets than other existing dimension reduction methods.Keywords: single-cell RNA-seq, minimum volume embedding, visualization, accelerated proximal gradient method
Procedia PDF Downloads 2285654 Induction of HIV-1 Resistance: The New Approaches Based on Gene Modification and Stem Cell Engineering
Authors: Alieh Farshbaf
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Introduction: Current anti-retroviral drugs have some restrictions for treatment of HIV-1 infection. The efficacy of retroviral drugs is not same in different infected patients and the virus rebound from latent reservoirs after stopping them. Recently, the engineering of stem cells and gene therapy provide new approaches to eliminate some drug problems by induction of resistance to HIV-1. Literature review: Up to now, AIDS-restriction genes (ARGs) were suitable candidate for gene and cell therapies, such as cc-chemokine receptor-5 (CCR5). In this manner, CCR5 provide effective cure in Berlin and Boston patients by inducing of HIV-1 resistance with allogeneic stem cell transplantation. It is showed that Zinc Finger Nuclease (ZFN) could induce HIV-1 resistance in stem cells of infected patients by homologous recombination or non-end joining mechanism and eliminate virus loading after returning the modified cells. Then, gene modification by HIV restriction factors, as TRIM5, introduced another gene candidate for HIV by interfering in infection process. These gene modifications/editing provided by stem cell futures that improve treatment in refractory disease such as HIV-1. Conclusion: Although stem cell transplantation has some complications, but in compare to retro-viral drugs demonstrated effective cure by elimination of virus loading. On the other hand, gene therapy is cost-effective for an infected patient than retroviral drugs payment in a person life-long. The results of umbilical cord blood stem cell transplantation showed that gene and cell therapy will be applied easier than previous treatment of AIDS with high efficacy.Keywords: stem cell, AIDS, gene modification, cell engineering
Procedia PDF Downloads 3025653 Isolation and Classification of Red Blood Cells in Anemic Microscopic Images
Authors: Jameela Ali Alkrimi, Abdul Rahim Ahmad, Azizah Suliman, Loay E. George
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Red blood cells (RBCs) are among the most commonly and intensively studied type of blood cells in cell biology. The lack of RBCs is a condition characterized by lower than normal hemoglobin level; this condition is referred to as 'anemia'. In this study, a software was developed to isolate RBCs by using a machine learning approach to classify anemic RBCs in microscopic images. Several features of RBCs were extracted using image processing algorithms, including principal component analysis (PCA). With the proposed method, RBCs were isolated in 34 second from an image containing 18 to 27 cells. We also proposed that PCA could be performed to increase the speed and efficiency of classification. Our classifier algorithm yielded accuracy rates of 100%, 99.99%, and 96.50% for K-nearest neighbor (K-NN) algorithm, support vector machine (SVM), and neural network ANN, respectively. Classification was evaluated in highly sensitivity, specificity, and kappa statistical parameters. In conclusion, the classification results were obtained for a short time period with more efficient when PCA was used.Keywords: red blood cells, pre-processing image algorithms, classification algorithms, principal component analysis PCA, confusion matrix, kappa statistical parameters, ROC
Procedia PDF Downloads 4055652 Evaluation of Promoter Hypermethylation in Tissue and Blood of Non-Small Cell Lung Cancer Patients and Association with Survival
Authors: Ashraf Ali, Kriti Upadhyay, Puja Sohal, Anant Mohan, Randeep Guleria
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Background: Gene silencing by aberrant promoter hypermethylation is common in lung cancer and is an initiating event in its development. Aim: To evaluate the gene promoter hypermethylation frequency in serum and tissue of lung cancer patients. Method: 95 newly diagnosed untreated advance stage lung cancer patients and 50 cancer free matched controls were studied. Bisulfite modification of tissue and serum DNA was done; modified DNA was used as a template for methylation-specific PCR analysis. Survival was assessed for one year. Results: Of 95 patients, 82% were non-small cell lung cancer (34% squamous cell carcinoma, 34% non-small cell lung cancer and 14% adenocarcinoma) and 18% were small cell lung cancer. Biopsy revealed that tissue of 89% and 75% of lung cancer patients and 85% and 52% of controls had promoter hypermethylated for MGMT (p=0.35) and p16(p<0.001) gene, respectively. In serum, 33% and 49% of lung cancer patients and 28% and 43% controls were positive for MGMT and p16 gene. No significant correlation was found between survival and clinico-pathological parameters. Conclusion: High gene promoter methylation frequency of p16 gene in tissue biopsy may be linked with early stages of carcinogenesis. Appropriate follow-up is required for confirmation of this finding.Keywords: lung cancer, MS- PCR, methylation, molecular biology
Procedia PDF Downloads 1955651 Lead in The Blood and Hypertension in Indonesia: A Systematic Review
Authors: Ainia Nurul Aqida
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Lead is one of the sources of air pollution. The use of lead on motor vehicle fuels resulted in the increasing contamination of lead in the air. The polluted air that has been inhaled by many people, especially guards and sellers of retail gasoline filling stations. The impact is increased levels of lead in blood. One result is an increase in blood pressure that causes hypertension. This research would like to know the relationship between blood lead levels in the incidence of hypertension in Indonesia. The method used in this study is a systematic review of the three journals have been published in the year 2007 to the year 2010 with the total sample is 312 samples. Odd ratio values obtained in the first article was OR = 6.50 pvalue = 0.000, CI = 95 % (2.89 to 14.60), and the second article was obtained OR 2.619 (95 % CI: 0.944 to 7.625) pvalue = 0.028, and the third article was obtained 0.002 r = 0.324 R2 = 10.5 %. Over all, there is a relationship between blood lead levels with the incidence of hypertension in Indonesia.Keywords: lead, blood, air pollution, hypertension
Procedia PDF Downloads 3245650 Monitoring Blood Pressure Using Regression Techniques
Authors: Qasem Qananwah, Ahmad Dagamseh, Hiam AlQuran, Khalid Shaker Ibrahim
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Blood pressure helps the physicians greatly to have a deep insight into the cardiovascular system. The determination of individual blood pressure is a standard clinical procedure considered for cardiovascular system problems. The conventional techniques to measure blood pressure (e.g. cuff method) allows a limited number of readings for a certain period (e.g. every 5-10 minutes). Additionally, these systems cause turbulence to blood flow; impeding continuous blood pressure monitoring, especially in emergency cases or critically ill persons. In this paper, the most important statistical features in the photoplethysmogram (PPG) signals were extracted to estimate the blood pressure noninvasively. PPG signals from more than 40 subjects were measured and analyzed and 12 features were extracted. The features were fed to principal component analysis (PCA) to find the most important independent features that have the highest correlation with blood pressure. The results show that the stiffness index means and standard deviation for the beat-to-beat heart rate were the most important features. A model representing both features for Systolic Blood Pressure (SBP) and Diastolic Blood Pressure (DBP) was obtained using a statistical regression technique. Surface fitting is used to best fit the series of data and the results show that the error value in estimating the SBP is 4.95% and in estimating the DBP is 3.99%.Keywords: blood pressure, noninvasive optical system, principal component analysis, PCA, continuous monitoring
Procedia PDF Downloads 1615649 Cytotoxic Activity of Extracts from Hibiscus sabdariffa Leaves against Women’s Cancer Cell Lines
Authors: Patsorn Worawattananutai, Srisopa Ruangnoo, Arunporn Itharat
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Hibiscus sabdariffa (HS) leaves are vegetables which are extensively used as blood tonic and laxatives in Thai traditional medicine. They are popularly used as healthy sour soup for prevention of chronic diseases such as cancer. Therefore, the cytotoxic activity of different extracts of fresh and dried Hibiscus sabdariffa leaves were investigated via the sulforhodamine B (SRB) assay against three types of women’s cancer cell lines, namely the human cervical adenocarcinoma cell line (HeLa), the human ovarian adenocarcinoma cell line (SKOV-3), and the human breast adenocarcinoma cell line (MCF-7). Extraction methods were squeezing, boiling with water and maceration with 95% or 50% ethanol. The 95% ethanolic extracts of Hibiscus sabdariffa dry leaves (HSDE95) showed the highest cytotoxicity against all types of women’s cancer cell lines with the IC50 values in range 7.51±0.33 to 12.13±1.85 µg/ml. Its IC50 values against SKOV-3, HeLa and MCF-7 were 7.51±0.33, 9.44±1.41 and 12.13±1.85 µg/ml, respectively. In these results, this extract can be classified as “active” according to the NCI guideline which indicated that IC50 values of the active cytotoxic plant extracts have to be beneath 20 µg/ml. Thus, HSDE95 was concluded to be a potent cytotoxic drug for all women’s cancer cells. This extract should be further investigated to isolate active compounds against women’s cancer cells.Keywords: breast adenocarcinoma, cervical adenocarcinoma, cytotoxic activity, Hibiscus sabdariffa, ovarian adenocarcinoma
Procedia PDF Downloads 6025648 Evaluation of Different Anticoagulant Effects on Flow Properties of Human Blood Using Falling Needle Rheometer
Authors: Hiroki Tsuneda, Takamasa Suzuki, Hideki Yamamoto, Kimito Kawamura, Eiji Tamura, Katharina Wochner, Roberto Plasenzotti
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Flow property of human blood is one of the important factors on the prevention of the circulatory condition such as a high blood pressure, a diabetes mellitus, and a cardiac infarction. However, the measurement of flow property of human blood, especially blood viscosity, is not so easy, because of their coagulation or aggregation behaviors after taking a sample from blood vessel. In the experiment, some kinds of anticoagulant were added into the human blood to avoid its solidification. Anticoagulant used in the blood test has been chosen for each purpose of blood test, for anticoagulant effect on blood is different mechanism for each. So that, there is a problem that the evaluation of measured blood property with different anticoagulant is so difficult. Therefore, it is so important to make clear the difference of anticoagulant effect on the blood property. In the previous work, a compact-size falling needle rheometer (FNR) has been developed in order to measure the flow property of human blood such as a flow curve, an apparent viscosity. It was found that FNR system can apply to a rheometer or a viscometry for various experimental conditions for not only human blood but also mammalians blood. In this study, the measurements of human blood viscosity with different anticoagulant (EDTA and Heparin) were carried out using newly developed FNR system. The effect of anticoagulant on blood viscosity was also tested by using the standard liquid for each. The accuracy on the viscometry was also tested by using the standard liquid for calibrating materials (JS-10, JS-20) and observed data have satisfactory agreement with reference data around 1.0% at 310K. The flow curve of six males and females with different anticoagulant were measured using FNR. In this experiment, EDTA and Heparin were chosen as anticoagulant for blood. Heparin can inhibit the coagulation of human blood by activating the body of anti-thrombin. To examine the effect of human blood viscosity on anticoagulant, flow curve was measured at high shear rate (>350s-1), and apparent viscosity of each person were determined with different anticoagulant. The apparent viscosity of human blood with heparin was 2%-9% higher than that with EDTA. However, the difference of blood viscosity for two anticoagulants for same blood was different for each. Further discussion, we need the consideration of effect on other physical property, such as cellular component and plasma component.Keywords: falling-needle rheometer, human blood, viscosity, anticoagulant
Procedia PDF Downloads 4425647 Blood Pressure and Anthropometric Measurements: A Correlational Study
Authors: Abdul-Monim Batiha, Manar AlAzzam, Mohammed ALBashtawy, Loai Tawalbeh, Ahmad Tubaishat, Fadwa N. Alhalaiqa
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Background: Obesity is the major modifiable risk factor for many chronic illnesses especially high blood pressure. Objectives: To evaluate the relationship between anthropometric indices and high blood pressure, and which one was most strongly correlated with high blood pressure in Jordanian population. Methods: A cross-sectional study was conducted with a total 622 students and workers from three Jordanian universities. Results: Nearly half of the participant are overweight (34.7%) and obese (15.4%) and hypertension was detected among 138 (22.2%) of the participants. Linear correlation was significant (p<0.01) between both systolic blood pressure and diastolic blood pressure for all anthropometric indices, except for A body shape index and diastolic blood pressure was significant at p< 0.05. Stepwise multiple linear regression analysis was used to assess the influence of age and anthropometric measurements. Conclusions: The waist circumference was the only independent predictor of hypertension, showing that this simple measurement may be an importance marker of high blood pressure in Jordanian population.Keywords: anthropometric indices, Jordan, blood pressure, cross-sectional study, obesity, hypertension, waist circumference
Procedia PDF Downloads 2885646 Up-Regulation of SCUBE2 Expression in Co-Cultures of Human Mesenchymal Stem Cell and Breast Cancer Cells
Authors: Hirowati Ali, Aisyah Ellyanti, Dewi Rusnita, Septelia Inawati Wanandi
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Stem cell has been known for its potency to be differentiated in many cells. Recently stem cell has been used for many treatment of degenerative medicine. It is still controversy whether stem cell can be used for therapy or these cells can activate cancer stem cell. SCUBE2 is a novel secreted and membrane-anchored protein which has been reported to its role in better prognosis and inhibition of cancer cell proliferation. Our study aims to observe whether stem cell can up-regulate SCUBE2 gene in MCF7 breast cancer cell line. We used in vitro study using MCF-7 cell treated with stem cell derived from placenta Wharton's jelly which has been known for its stemness and widely used. Our results showed that MCF-7 cell line grows up rapidly in 6-well culture dish. Stem cell was cultured in 6-well dish. After 50%-60% MCF-7 confluence, we co-cultured these cells with stem cells for 24 hours and 48 hours. We hypothesize SCUBE2 gene which is previously known for its higher expression in better prognosis of breast cancer, is up-regulated after stem cells addition in MCF7 culture dishes.Keywords: breast cancer cells, inhibition of cancer cells, mesenchymal stem cells, SCUBE2
Procedia PDF Downloads 3405645 Effect of Far Infrared and Endothelial Cell Growth Supplement on Human Umbilical Vascular Endothelial Cells
Authors: Ming-Tzu Tsai, Jui-Ting Hsu, Chia-Chieh Lin, Feng-Tsai Chiang, Cheng-Chin Huang
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Far infrared (FIR), an invisible and short electromagnetic waves ranges from 6-14 μm also defines as the “growth ray.” Although the mechanism of FIR is still unknown, most data have suggested that FIR could accelerate the skin microcirculation by elevating the blood flow and nitric-oxide (NO) synthesis. In this present work, the effect of FIR irradiation and endothelial cell growth supplement (ECGS) on human umbilical vascular endothelial cells (HUVECs) was evaluated. To understand whether the cell viability and NO production of HUVECs affected by NO, cells with/without ECGS were treated in the presence or absence of L-NAME, an eNOS inhibitor. For FIR exposure, FIR-emitted ceramic powders consisted of a variety of well-mixed metal oxides were developed. The results showed that L-NAME did had a strong effect on the inhibition of NO production, especially in the ECGS-treated group. However, the cell viability of each group was rarely affected in the presence of L-NAME. Cells with the incubation of ECGS showed much higher cell viability compared to the control. Moreover, NO production of HUVECs exposed to FIR irradiation was significantly inhibited in the presence of L-NAME. It suggested that NO could play a role modulating the downstream signals of HUVECs during FIR exposure.Keywords: far-infrared irradiation (FIR), nitric oxide (NO), endothelial nitric oxide synthase (eNOS), endothelial cell growth supplement (ECGS)
Procedia PDF Downloads 4295644 Human Immuno-Deficiency Virus Co-Infection with Hepatitis B Virus and Baseline Cd4+ T Cell Count among Patients Attending a Tertiary Care Hospital, Nepal
Authors: Soma Kanta Baral
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Background: Since 1981, when the first AIDS case was reported, worldwide, more than 34 million people have been infected with HIV. Almost 95 percent of the people infected with HIV live in developing countries. As HBV & HIV share similar routes of transmission by sexual intercourse or drug use by parenteral injection, co-infection is common. Because of the limited access to healthcare & HIV treatment in developing countries, HIV-infected individuals are present late for care. Enumeration of CD4+ T cell count at the time of diagnosis has been useful to initiate the therapy in HIV infected individuals. The baseline CD4+ T cell count shows high immunological variability among patients. Methods: This prospective study was done in the serology section of the Department of Microbiology over a period of one year from august 2012 to July 2013. A total of 13037 individuals subjected for HIV test were included in the study comprising of 4982 males & 8055 females. Blood sample was collected by vein puncture aseptically with standard operational procedure in clean & dry test-tube. All blood samples were screened for HIV as described by WHO algorithm by Immuno-chromatography rapid kits. Further confirmation was done by biokit ELISA method as per the manufacturer’s guidelines. After informed consent, HIV positive individuals were screened for HBsAg by Immuno-chromatography rapid kits (Hepacard). Further confirmation was done by biokit ELISA method as per the manufacturer’s guidelines. EDTA blood samples were collected from the HIV sero-positive individuals for baseline CD4+ T count. Then, CD4+ T cells count was determined by using FACS Calibur Flow Cytometer (BD). Results: Among 13037 individuals screened for HIV, 104 (0.8%) were found to be infected comprising of 69(66.34%) males & 35 (33.65%) females. The study showed that the high infection was noted in housewives (28.7%), active age group (30.76%), rural area (56.7%) & in heterosexual route (80.9%) of transmission. Out of total HIV infected individuals, distribution of HBV co-infection was found to be 6(5.7%). All co- infected individuals were married, male, above the age of 25 years & heterosexual route of transmission. Baseline CD4+ T cell count of HIV infected patient was found higher (mean CD4+ T cell count; 283cells/cu.mm) than HBV co-infected patients (mean CD4+ T cell count; 91 cells/cu.mm). Majority (77.2%) of HIV infected & all co-infected individuals were presented in our center late (CD4+ T cell count;< 350/cu. mm) for diagnosis and care. Majority of co- infected 4 (80%) were late presented with advanced AIDS stage (CD4+ count; <200/cu.mm). Conclusions: The study showed a high percentage of HIV sero-positive & co- infected individuals. Baseline CD4+ T cell count of majority of HIV infected individuals was found to be low. Hence, more sustained and vigorous awareness campaigns & counseling still need to be done in order to promote early diagnosis and management.Keywords: HIV/AIDS, HBsAg, co-infection, CD4+
Procedia PDF Downloads 2175643 In vitro Regeneration of Neural Cells Using Human Umbilical Cord Derived Mesenchymal Stem Cells
Authors: Urvi Panwar, Kanchan Mishra, Kanjaksha Ghosh, ShankerLal Kothari
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Background: Day-by-day the increasing prevalence of neurodegenerative diseases have become a global issue to manage them by medical sciences. The adult neural stem cells are rare and require an invasive and painful procedure to obtain it from central nervous system. Mesenchymal stem cell (MSCs) therapies have shown remarkable application in treatment of various cell injuries and cell loss. MSCs can be derived from various sources like adult tissues, human bone marrow, umbilical cord blood and cord tissue. MSCs have similar proliferation and differentiation capability, but the human umbilical cord-derived mesenchymal stem cells (hUCMSCs) are proved to be more beneficial with respect to cell procurement, differentiation to other cells, preservation, and transplantation. Material and method: Human umbilical cord is easily obtainable and non-controversial comparative to bone marrow and other adult tissues. The umbilical cord can be collected after delivery of baby, and its tissue can be cultured using explant culture method. Cell culture medium such as DMEMF12+10% FBS and DMEMF12+Neural growth factors (bFGF, human noggin, B27) with antibiotics (Streptomycin/Gentamycin) were used to culture and differentiate mesenchymal stem cells into neural cells, respectively. The characterisations of MSCs were done with Flow Cytometer for surface markers CD90, CD73 and CD105 and colony forming unit assay. The differentiated various neural cells will be characterised by fluorescence markers for neurons, astrocytes, and oligodendrocytes; quantitative PCR for genes Nestin and NeuroD1 and Western blotting technique for gap43 protein. Result and discussion: The high quality and number of MSCs were isolated from human umbilical cord via explant culture method. The obtained MSCs were differentiated into neural cells like neurons, astrocytes and oligodendrocytes. The differentiated neural cells can be used to treat neural injuries and neural cell loss by delivering cells by non-invasive administration via cerebrospinal fluid (CSF) or blood. Moreover, the MSCs can also be directly delivered to different injured sites where they differentiate into neural cells. Therefore, human umbilical cord is demonstrated to be an inexpensive and easily available source for MSCs. Moreover, the hUCMSCs can be a potential source for neural cell therapies and neural cell regeneration for neural cell injuries and neural cell loss. This new way of research will be helpful to treat and manage neural cell damages and neurodegenerative diseases like Alzheimer and Parkinson. Still the study has a long way to go but it is a promising approach for many neural disorders for which at present no satisfactory management is available.Keywords: bone marrow, cell therapy, explant culture method, flow cytometer, human umbilical cord, mesenchymal stem cells, neurodegenerative diseases, neuroprotective, regeneration
Procedia PDF Downloads 2025642 Efficient Pre-Processing of Single-Cell Assay for Transposase Accessible Chromatin with High-Throughput Sequencing Data
Authors: Fan Gao, Lior Pachter
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The primary tool currently used to pre-process 10X Chromium single-cell ATAC-seq data is Cell Ranger, which can take very long to run on standard datasets. To facilitate rapid pre-processing that enables reproducible workflows, we present a suite of tools called scATAK for pre-processing single-cell ATAC-seq data that is 15 to 18 times faster than Cell Ranger on mouse and human samples. Our tool can also calculate chromatin interaction potential matrices, and generate open chromatin signal and interaction traces for cell groups. We use scATAK tool to explore the chromatin regulatory landscape of a healthy adult human brain and unveil cell-type specific features, and show that it provides a convenient and computational efficient approach for pre-processing single-cell ATAC-seq data.Keywords: single-cell, ATAC-seq, bioinformatics, open chromatin landscape, chromatin interactome
Procedia PDF Downloads 1565641 Ultrasensitive Hepatitis B Virus Detection in Blood Using Nano-Porous Silicon Oxide: Towards POC Diagnostics
Authors: N. Das, N. Samanta, L. Pandey, C. Roy Chaudhuri
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Early diagnosis of infection like Hep-B virus in blood is important for low cost medical treatment. For this purpose, it is desirable to develop a point of care device which should be able to detect trace quantities of the target molecule in blood. In this paper, we report a nanoporous silicon oxide sensor which is capable of detecting down to 1fM concentration of Hep-B surface antigen in blood without the requirement of any centrifuge or pre-concentration. This has been made possible by the presence of resonant peak in the sensitivity characteristics. This peak is observed to be dependent only on the concentration of the specific antigen and not on the interfering species in blood serum. The occurrence of opposite impedance change within the pores and at the bottom of the pore is responsible for this effect. An electronic interface has also been designed to provide a display of the virus concentration.Keywords: impedance spectroscopy, ultrasensitive detection in blood, peak frequency, electronic interface
Procedia PDF Downloads 4035640 Direct Assessment of Cellular Immune Responses to Ovalbumin with a Secreted Luciferase Transgenic Reporter Mouse Strain IFNγ-Lucia
Authors: Martyna Chotomska, Aleksandra Studzinska, Marta Lisowska, Justyna Szubert, Aleksandra Tabis, Jacek Bania, Arkadiusz Miazek
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Objectives: Assessing antigen-specific T cell responses is of utmost importance for the pre-clinical testing of prototype vaccines against intracellular pathogens and tumor antigens. Mainly two types of in vitro assays are used for this purpose 1) enzyme-linked immunospot (ELISpot) and 2) intracellular cytokine staining (ICS). Both are time-consuming, relatively expensive, and require manual dexterity. Here, we assess if a straightforward detection of luciferase activity in blood samples of transgenic reporter mice expressing a secreted Lucia luciferase under the transcriptional control of IFN-γ promoter parallels the sensitivity of IFNγ ELISpot assay. Methods: IFN-γ-LUCIA mouse strain carrying multiple copies of Lucia luciferase transgene under the transcriptional control of IFNγ minimal promoter were generated by pronuclear injection of linear DNA. The specificity of transgene expression and mobilization was assessed in vitro using transgenic splenocytes exposed to various mitogens. The IFN-γ-LUCIA mice were immunized with 50mg of ovalbumin (OVA) emulsified in incomplete Freund’s adjuvant three times every two weeks by subcutaneous injections. Blood samples were collected before and five days after each immunization. Luciferase activity was assessed in blood serum. Peripheral blood mononuclear cells were separated and assessed for frequencies of OVA-specific IFNγ-secreting T cells. Results: We show that in vitro cultured splenocytes of IFN-γ-LUCIA mice respond by 2 and 3 fold increase in secreted luciferase activity to T cell mitogens concanavalin A and phorbol myristate acetate, respectively but fail to respond to B cell-stimulating E.coli lipopolysaccharide. Immunization of IFN-γ-LUCIA mice with OVA leads to over 4 fold increase in luciferase activity in blood serum five days post-immunization with a barely detectable increase in OVA-specific, IFNγ-secreting T cells by ELISpot. Second and third immunizations, further increase the luciferase activity and coincidently also increase the frequencies of OVA-specific T cells by ELISpot. Conclusions: We conclude that minimally invasive monitoring of luciferase secretions in blood serum of IFN-γ-LUCIA mice constitutes a sensitive method for evaluating primary and memory Th1 responses to protein antigens. As such, this method may complement existing methods for rapid immunogenicity assessment of prototype vaccines.Keywords: ELISpot, immunogenicity, interferon-gamma, reporter mice, vaccines
Procedia PDF Downloads 172