Excellence in Research and Innovation for Humanity

International Science Index

Commenced in January 1999 Frequency: Monthly Edition: International Abstract Count: 40434

Biomedical and Biological Engineering

An Advanced Automated Brain Tumor Diagnostics Approach
During past few years, automation of tumor detection in magnetic resonance imaging (MRI) has become a challenging research area in the field of medical imaging. Automated tumor detection is required because there might be a shortage of skilled radiologists at a time of great need. In this study, an automated brain tumor detection and localization framework that can detect and localize brain tumors in magnetic resonance imaging are proposed. The aim of the study is to detect and to identify the tumor formation in the brain with using image processing techniques and Probabilistic Neural Network (PNN) method. In the experimental part of the study, 100 neuroimages are used to optimize our system, and 50 out-of-sample neuroimages are also used to test the approach. The preliminary results demonstrate the high classification accuracy for the neural network structure. Moreover, the preliminary results also motivate us to extend this framework to detect and localize the tumors in the other organs.
Alternative Epinephrine Injector to Combat Allergy Induced Anaphylaxis
One response during anaphylaxis is reduced blood pressure due to blood vessels relaxing and dilating. Epinephrine causes the blood vessels to constrict, which raises blood pressure to counteract the symptoms. When going through an allergic reaction, an Epinephrine injector is used to administer a shot of epinephrine intramuscularly. Epinephrine injectors have become an integral part of day-to-day life for people with allergies. Current Epinephrine injectors (EpiPen) are completely mechanical and have no sensors to monitor the vital signs of patients or give suggestions the optimal time for the shot. The EpiPens are also large and inconvenient to carry daily. The current price of an EpiPen is roughly 600$ for a pack of two. This makes carrying an EpiPen very expensive, especially when they need to be switched out when the epinephrine expires. This new design is in the form of a bracelet, which has the ability to inject epinephrine. The bracelet will be equipped with vital signs monitors that can aid the patient to sense the allergic reaction. The vital signs that would be of interest are blood pressure, heart rate and Electrodermal activity (EDA). The heart rate of the patient will be tracked by a photoplethysmograph (PPG) that is incorporated into the sensors. The heart rate is expected to increase during anaphylaxis. Blood pressure will be monitored through a radar sensor, which monitors the phase changes in electromagnetic waves as they reflect off of the blood vessel. EDA is under autonomic control. Allergen-induced anaphylaxis is caused by a release of chemical mediators from mast cells and basophils, thus changes the autonomic activity of the patient. So by measuring EDA, it will give the wearer an alert on how their autonomic nervous system is reacting. After the vital signs are collected, they will be sent to an application on a smartphone to be analyzed, which can then alert an emergency contact if the epinephrine injector on the bracelet is activated. Overall, this design creates a safer system by aiding the user in keeping track of their epinephrine injector, while making it easier to track their vital signs. Also, our design will be more affordable and more convenient to replace. Rather than replacing the entire product, only the needle and drug will be switched out and not the entire design.
Response of Planktonic and Aggregated Bacterial Cells to Water Disinfection with Photodynamic Inactivation
The interest in developing alternative techniques to obtain safe water, free from pathogens and hazardous substances, is growing in recent times. The photodynamic inactivation of microorganisms (PDI) is a promising ecologically-friendly and multi-target approach for water disinfection. It uses visible light as an energy source combined with a photosensitiser (PS) to transfer energy/electrons to a substrate or molecular oxygen generating reactive oxygen species, which cause cidal effects towards cells. PDI has mainly been used in clinical studies and investigations on its application to disinfect water is relatively recent. The majority of studies use planktonic cells. However, in their natural environments, bacteria quite often do not occur as freely suspended cells (planktonic) but in cell aggregates that are either freely floating or attached to surfaces as biofilms. Microbes can form aggregates and biofilms as a strategy to protect them from environmental stress. As aggregates, bacteria have a better metabolic function, they communicate more efficiently, and they are more resistant to biocide compounds than their planktonic forms. Among the bacteria that are able to form aggregates are members of the genus Pseudomonas, they are a very diverse group widely distributed in the environment. Pseudomonas species can form aggregates/biofilms in water and can cause particular problems in water distribution systems. The aim of this study was to evaluate the effectiveness of photodynamic inactivation in killing a range of planktonic cells including Escherichia coli DSM 1103, Staphylococcus aureus DSM 799, Shigella sonnei DSM 5570, Salmonella enterica and Pseudomonas putida DSM 6125, and aggregating cells of Pseudomonas fluorescens DSM 50090, Pseudomonas aeruginosa PAO1. The experiments were performed in glass Petri dishes, containing the bacterial suspension and the photosensitiser, irradiated with a multi-LED (wavelengths 430nm and 660nm) for different time intervals. The responses of the cells were monitored using the pour plate technique and confocal microscopy. The study showed that bacteria belonging to Pseudomonads group tend to be more tolerant to PDI. While E. coli, S. aureus, S. sonnei and S. enterica required a dosage ranging from 39.47 J/cm2 to 59.21 J/cm2 for a 5 log reduction, Pseudomonads needed a dosage ranging from 78.94 to 118.42 J/cm2, a higher dose being required when the cells aggregated.
Comparison of the Effects of Rod Types of Rigid Fixation Devices on the Loads in the Lumbar Spine: A Finite Element Analysis
We developed new design of rod of pedicle screw system that is beneficial in maintaining the spacing between the vertebrae and assessed the performance of the posterior fixation screw systems by numerical analysis according to the range of motion (flexion, extension, lateral bending, and axial rotation) of the vertebral column after inserting the pedicle screws. The simulation results showed that the conventional rod was the most low equivalent stress value among implant units in the case of flexion, extension and lateral bending of the vertebrae. In all cases except the torsional rotation, the results showed that the stress level of the single and double rounded rod exceeded about 30% to 70% compare to the conventional rod. Therefore, this product is not suitable for actual application in the field yet and it seems that product design optimization is necessary. Acknowledgement: This research was supported by the Ministry of Trade, Industry & Energy (MOTIE), Korea Institute for Advancement of Technology (KIAT) through the Encouragement Program for The Industries of Economic Cooperation Region.
Design and Implementation of a Wearable Artificial Kidney Prototype for Home Dialysis
Hemodialysis is a life-preserving treatment for a number of patients with kidney failure. The standard procedure of hemodialysis is three times a week during the hemodialysis procedure, the patient usually suffering from many inconvenient, exhausting feeling and effect on the heart and cardiovascular system are the most common signs. This paper provides a solution to reduce the previous problems by designing a wearable artificial kidney (WAK) taking in consideration a minimization the size of the dialysis machine. The WAK system consists of two circuits: blood circuit and dialysate circuit. The blood from the patient is filtered in the dialyzer before returning back to the patient. Several parameters using an advanced microcontroller and array of sensors. WAK equipped with visible and audible alarm system to aware the patients if there is any problem.
Distinguishing between Bacterial and Viral Infections Based on Peripheral Human Blood Tests Using Infrared Microscopy and Multivariate Analysis
Viral and bacterial infections are responsible for variety of diseases. These infections have similar symptoms like fever, sneezing, inflammation, vomiting, diarrhea and fatigue. Thus, physicians may encounter difficulties in distinguishing between viral and bacterial infections based on these symptoms. Bacterial infections differ from viral infections in many other important respects regarding the response to various medications and the structure of the organisms. In many cases, it is difficult to know the origin of the infection. The physician orders a blood, urine test, or 'culture test' of tissue to diagnose the infection type when it is necessary. Using these methods, the time that elapses between the receipt of patient material and the presentation of the test results to the clinician is typically too long ( > 24 hours). This time is crucial in many cases for saving the life of the patient and for planning the right medical treatment. Thus, rapid identification of bacterial and viral infections in the lab is of great importance for effective treatment especially in cases of emergency. Blood was collected from 50 patients with confirmed viral infection and 50 with confirmed bacterial infection. White blood cells (WBCs) and plasma were isolated and deposited on a zinc selenide slide, dried and measured under a Fourier transform infrared (FTIR) microscope to obtain their infrared absorption spectra. The acquired spectra of WBCs and plasma were analyzed in order to differentiate between the two types of infections. In this study, the potential of FTIR microscopy in tandem with multivariate analysis was evaluated for the identification of the agent that causes the human infection. The method was used to identify the infectious agent type as either bacterial or viral, based on an analysis of the blood components [i.e., white blood cells (WBC) and plasma] using their infrared vibrational spectra. The time required for the analysis and evaluation after obtaining the blood sample was less than one hour. In the analysis, minute spectral differences in several bands of the FTIR spectra of WBCs were observed between groups of samples with viral and bacterial infections. By employing the techniques of feature extraction with linear discriminant analysis (LDA), a sensitivity of ~92 % and a specificity of ~86 % for an infection type diagnosis was achieved. The present preliminary study suggests that FTIR spectroscopy of WBCs is a potentially feasible and efficient tool for the diagnosis of the infection type.
Fabrication of Optical Tissue Phantoms Simulating Human Skin and Their Application
Although various optical tissue phantoms (OTPs) simulating human skin have been actively studied, their completeness is unclear because skin tissue has the intricate optical property and complicated structure disturbing the optical simulation. In this study, we designed multilayer OTP mimicking skin structure, and fabricated OTP models simulating skin-blood vessel and skin pigmentation in the skin, which are useful in Biomedical optics filed. The OTPs were characterized with the optical property and the cross-sectional structure, and analyzed by using various optical tools such as a laser speckle imaging system, OCT and a digital microscope to show the practicality. The measured optical property was within 5% error, and the thickness of each layer was uniform within 10% error in micrometer scale.
Green Synthesis and Characterization of Zinc and Ferrous Nanoparticles for Their Potent Therapeutic Approach
Green nanotechnology is the most researched field in the current scenario. Herein we study the synthesis of Zinc and Ferrous nanoparticles using Moringa oleifera leaf extracts. Our protocol using established protocols heat treatment of plant extracts along with the solution of copper sulphate in the ratio of 1:1. The leaf extracts of Moringa oleifera were prepared in deionized water. Copper sulfate solution (1mM) was added to this, and the change in color of the solution was observed indicating the formation of Cu nanoparticles. The as biosynthesized Cu nanoparticles were characterized with the help of Scanning Electron Microscopy (SEM), and Fourier Transforms Infrared Spectroscopy (FTIR). It was observed that the leaf extracts of Moringa oleifera can reduce copper ions into copper nanoparticles within 8 to 10 min of reaction time. The method thus can be used for rapid and eco-friendly biosynthesis of stable copper nanoparticles. Further, we checked their antimicrobial and antioxidant potential, and it was observed that maximum antioxidant activity was observed for the particles prepared using the heating method. The maximum antibacterial activity was observed in Streptomyces grisveus particles and in Triochoderma Reesei for the maximum antifungal activity. At present, we are engaged in studying the anti-inflammatory activities of these as prepared nanoparticles.
Comprehensive Analysis of Electrohysterography Signal Features in Term and Preterm Labor
Premature birth, defined as birth before 37 completed weeks of gestation is a leading cause of neonatal morbidity and mortality and has long-term adverse consequences for health. It has recently been reported that the worldwide preterm birth rate is around 10%. The existing measurement techniques for diagnosing preterm delivery include tocodynamometer, ultrasound and fetal fibronectin. However, they are subjective, or suffer from high measurement variability and inaccurate diagnosis and prediction of preterm labor. Electrohysterography (EHG) method based on recording of uterine electrical activity by electrodes attached to maternal abdomen, is a promising method to assess uterine activity and diagnose preterm labor. The purpose of this study is to analyze the difference of EHG signal features between term labor and preterm labor. Free access database was used with 300 signals acquired in two groups of pregnant women who delivered at term (262 cases) and preterm (38 cases). Among them, EHG signals from 38 term labor and 38 preterm labor were preprocessed with band-pass Butterworth filters of 0.08–4Hz. Then, EHG signal features were extracted, which comprised classical time domain description including root mean square and zero-crossing number, spectral parameters including peak frequency, mean frequency and median frequency, wavelet packet coefficients, autoregression (AR) model coefficients, and nonlinear measures including maximal Lyapunov exponent, sample entropy and correlation dimension. Their statistical significance for recognition of two groups of recordings was provided. The results showed that mean frequency of preterm labor was significantly smaller than term labor (p < 0.05). 5 coefficients of AR model showed significant difference between term labor and preterm labor. The maximal Lyapunov exponent of early preterm (time of recording < the 26th week of gestation) was significantly smaller than early term. The sample entropy of late preterm (time of recording > the 26th week of gestation) was significantly smaller than late term. There was no significant difference for other features between the term labor and preterm labor groups. Any future work regarding classification should therefore focus on using multiple techniques, with the mean frequency, AR coefficients, maximal Lyapunov exponent and the sample entropy being among the prime candidates. Even if these methods are not yet useful for clinical practice, they do bring the most promising indicators for the preterm labor.
Prevalence and Associated Factors of Stunting among 6-59 Months Children in Pastoral Community of Korahay Zone, Somali Regional State, Ethiopia 2016
Background: Stunting is one of the most important public health problems in Ethiopia with an estimated 44.4% of children less than five years of age are stunted. Thus, this study aimed to assess prevalence and associated factors of stunting among 6-59 months children in pastoral community of Korahay Zone, Somali Regional State, Ethiopia. Objective of the study: To assess prevalence and associated factors of stunting among 6-59 months children in pastoral community of Korahay Zone, Somali Regional State, Ethiopia, 2016. Methods: Community based cross sectional study design was done among 770 children in pastoral community of Korahay Zone. Systematic sampling techniques were used to select households and took child mother pair from each selected households. Data was collected using pre-tested and structured questionnaire. Odds ratio with 95% confidence interval was used to assess level of significance. Result: Prevalence of stunting among 6-59 months age children was 31.9%. Sex (AOR: 1.47, 95%CI 1.02, 2.11), age (AOR: 2.10, 95%CI 1.16, 3.80), maternal education (AOR: 3.42, 95%CI 1.58, 7.41), maternal occupation (AOR: 3.10, 95%CI 1.85, 5.19), monthly income (AOR: 1.47, 95%CI 1.03, 2.09), PNC visits (AOR: 1.59, 95%CI 1.07, 2.37), source of water (AOR: 3.41, 95%CI 1.96, 5.93), toilet availability (AOR: 1.71, 95%CI 1.13, 2.58), first milk feeding (AOR: 3.37, 95%CI 2.27, 5.02) and bottle feeding (AOR: 2.07, 95%CI 1.34, 3.18) were significant predictors of stunting. Conclusion and recommendations: Prevalence of stunting among 6-59 months children was high 31.9%. Lack maternal education, not feeding first milk, unsafe water supply, absence toilet availability and bottle feeding can increase the risk of stunting. So, educating mothers on child feeding practice, sanitation and important of first milk can reduce stunting.
A Constitutive Model of Ligaments and Tendons Accounting for Fiber-Matrix Interaction
In this study, a new constitutive model is developed to describe the hyperelastic behavior of collagenous tissues with a parallel arrangement of collagen fibers such as ligaments and tendons. The model is formulated using a continuum approach incorporating the structural changes of the main tissue components: collagen fibers, proteoglycan-rich matrix and fiber-matrix interaction. The mechanical contribution of the interaction between the fibers and the matrix is simply expressed by a coupling term. The structural change of the collagen fibers is incorporated in the constitutive model to describe the activation of the fibers under tissue straining. Finally, the constitutive model can easily describe the stress-stretch nonlinearity which occurs when a ligament/tendon is axially stretched. This study shows that the interaction between the fibers and the matrix contributes to the mechanical tissue response. Therefore, the model may lead to a better understanding of the physiological mechanisms of ligaments and tendons under axial loading.
Connecting MRI Physics to Glioma Microenvironment: Comparing Simulated T2-Weighted MRI Models of Fixed and Expanding Extracellular Space
Glioblastoma Multiforme (GBM), the most common primary brain tumor, often presents with hyperintensity on T2-weighted or T2-weighted fluid attenuated inversion recovery (T2/FLAIR) magnetic resonance imaging (MRI). This hyperintensity corresponds with vasogenic edema, however there are likely many infiltrating tumor cells within the hyperintensity as well. While MRIs do not directly indicate tumor cells, MRIs do reflect the microenvironmental water abnormalities caused by the presence of tumor cells and edema. The inherent heterogeneity and resulting MRI features of GBMs complicate assessing disease response. To understand how hyperintensity on T2/FLAIR MRI may correlate with edema in the extracellular space (ECS), a multi-compartmental MRI signal equation which takes into account tissue compartments and their associated volumes with input coming from a mathematical model of glioma growth that incorporates edema formation was explored. The reasonableness of two possible extracellular space schema was evaluated by varying the T2 of the edema compartment and calculating the possible resulting T2s in tumor and peripheral edema. In the mathematical model, gliomas were comprised of vasculature and three tumor cellular phenotypes: normoxic, hypoxic, and necrotic. Edema was characterized as fluid leaking from abnormal tumor vessels. Spatial maps of tumor cell density and edema for virtual tumors were simulated with different rates of proliferation and invasion and various ECS expansion schemes. These spatial maps were then passed into a multi-compartmental MRI signal model for generating simulated T2/FLAIR MR images. Individual compartments’ T2 values in the signal equation were either from literature or estimated and the T2 for edema specifically was varied over a wide range (200 ms – 9200 ms). T2 maps were calculated from simulated images. T2 values based on simulated images were evaluated for regions of interest (ROIs) in normal appearing white matter, tumor, and peripheral edema. The ROI T2 values were compared to T2 values reported in literature. The expanding scheme of extracellular space is had T2 values similar to the literature calculated values. The static scheme of extracellular space had a much lower T2 values and no matter what T2 was associated with edema, the intensities did not come close to literature values. Expanding the extracellular space is necessary to achieve simulated edema intensities commiserate with acquired MRIs.
Electrochemically Reduced Graphene Oxide Modified Boron-Doped Diamond Paste Electrode on Paper-Based Analytical Device for Simultaneous Determination of Norepinephrine and Serotonin
In this work, we demonstrate a novel electrochemically reduced graphene oxide (ERGO) modified boron-doped diamond paste (BDDP) electrode on paper-based analytical devices (PADs) for simultaneous determination of norepinephrine (NE) and serotonin (5-HT). The BDD paste electrode was easily constructed by filling BDD paste in small channels, which made in transparency film sheets using a CO₂ laser etching 
system. The counter and reference electrodes were fabricated on paper by in-house screen-printing and then combined with BDD paste microelectrode. The electrochemical characterization of the device was investigated by cyclic voltammetry (CV). Differential pulse voltammetry (DPV) was employed for the simultaneous determination of NE and 5-HT. The ERGO-modified BDDP electrode displayed excellent electrocatalytic activities toward the oxidation of NE and 5-HT and strong function for resolving the overlapping voltammetric responses of NE and 5-HT into two well-defined voltammetric peaks. This device was capable of simultaneously detecting NE and 5-HT in wide concentration ranges and with a low limit of detections. In addition, it has the advantages in terms of ease of use, low cost, and disposability.
Precise Spatially Selective Photothermolysis Skin Treatment by Multiphoton Absorption
Conventional laser treatment of skin diseases and cosmetic surgery is based on the principle of one-photon absorption selective photothermolysis which relies strongly on the difference in the light absorption between the therapeutic target and its surrounding tissue. However, when the difference in one-photon absorption is not sufficient, collateral damage would occur due to indiscriminate and nonspecific tissue heating. To overcome this problem, we developed a spatially selective photothermolysis method based on multiphoton absorption in which the heat generation is restricted to the focal point of a tightly focused near-infrared femtosecond laser beam aligned with the target of interest. A multimodal optical microscope with co-registered reflectance confocal imaging (RCM), two-photon fluorescence imaging (TPF), and second harmonic generation imaging (SHG) capabilities was used to perform and monitor the spatially selective photothermolysis. Skin samples excised from the shaved backs of euthanized NODSCID mice were used in this study. Treatments were performed by focusing and scaning the laser beam in the dermis with a 50µm×50µm target area. Treatment power levels of 200 mW to 400 mW and modulated pulse trains of different duration and period were experimented. Different treatment parameters achieved different degrees of spatial confinement of tissue alterations as visualized by 3-D RCM/TPF/SHG imaging. At 200 mW power level, 0.1 s pulse train duration, 4.1 s pulse train period, the tissue damage was found to be restricted precisely to the 50µm×50µm×10µm volume, where the laser focus spot had scanned through. The overlying epidermis/dermis tissue and the underneath dermis tissue were intact although there was light passing through these regions.
Barnard Feature Point Detector for Low-Contractperiapical Radiography Image
In dental clinics, the dentists use the periapical radiography image to assess the effectiveness of endodontic treatment of teeth with chronic apical periodontitis. Periapical radiography images are taken at different times to assess alveolar bone variation before and after the root canal treatment, and furthermore to judge whether the treatment was successful. Current clinical assessment of apical tissue recovery relies only on dentist personal experience. It's hard to have the same standard and objective interpretations due to the dentist or radiologist personal background and knowledge. If periapical radiography images at the different time could be registered well, the endodontic treatment could be evaluated. In the image registration area, it is necessary to assign representative control points to the transformation model for good performances of registration results. However, detection of representative control points (feature points) on periapical radiography images is generally very difficult. Regardless of which traditional detection methods are practiced, sufficient feature points may not be detected due to the low-contrast characteristics of the x-ray image. Barnard detector is an algorithm for feature point detection based on grayscale value gradients, which can obtain sufficient feature points in the case of gray-scale contrast is not obvious. However, the Barnard detector would detect too many feature points, and they would be too clustered. This study uses the local extrema of clustering feature points and the suppression radius to overcome the problem, and compared different feature point detection methods. In the preliminary result, the feature points could be detected as representative control points by the proposed method.
Accelerated Molecular Simulation: A Convolution Approach
Computational Drug Design is often based on molecular dynamics simulations of molecular systems. Molecular dynamics can be used to simulate, e.g., the binding and unbinding event of a small drug-like molecule with regard to the active site of an enzyme or a receptor. However, the time-scale of the overall binding event is many orders of magnitude longer than the time-scale of simulation. Thus, there is a need to speed-up molecular simulations. In order to speed up simulations, the molecular dynamics trajectories have to be 'steared' out of local minimizers of the potential energy surface – the so-called metastabilities – of the molecular system. Increasing the kinetic energy (temperature) is one possibility to accelerate simulated processes. However, with temperature, the entropy of the molecular system increases, too. But this kind 'stearing' is not directed enough to stear the molecule out of the minimum toward the saddle point. In this article, we give a new mathematical idea, how a potential energy surface can be changed in such a way, that entropy is kept under control while the trajectories are still steared out of the metastabilities. In order to compute the unsteared transition behaviour based on a steared simulation, we propose to use extrapolation methods. In the end we mathematically show, that our method accelerates the simulations along the direction, in which the curvature of the potential energy surface changes the most, i.e., from local minimizers towards saddle points.
Screening of Thyroid Stimulating Hormone Using Paper-Based Lateral Flow Device
A paper-based lateral flow device for screening thyroid stimulating hormone (TSH) is reported. A sandwich immunoassay was performed using two mouse monoclonal TSH antibodies (anti-hTSH 5403 and 5404) as immobilized and labeled antibodies for capturing TSH samples. Test (anti-hTSH 5403) and control (goat anti-Mouse IgG) lines were fabricated on nitrocellulose membrane (NCM) using ballpoint pen printed with a speed of 3 cm/s and thickness setting of 1. The novel gold nanoparticles europium complex (AuNPs@Eu) was used as fluorescence label compared to conventional AuNPs label. The results obtained with this device can be visually assessed by the naked eyes and under UV hand lamps, and quantitative analysis can be performed using the ImageJ program. The limit of detection (LOD) under UV hand lamps (0.1 µIU/mL) provided 50-fold greater sensitivity than AuNPs (5 µIU/mL), which is suitable for both hypothyroidism and hyperthyroidism screening within 30 min. A linear relationship between the red intensity and the logarithmic concentrations of TSH was observed with a good correlation (R²=0.992). Furthermore, the device can be effectively applied for screening TSH in the spiked human serum with recovery range of 96.80-104.45% and RSD of 2.18-3.63%. Therefore, the developed device is an alternative method for TSH screening which provides a lot of advantages including low cost, short time analysis, ease of use, disposability, portability, and on-site measurement.
Production of Recombinant Human Serum Albumin in Escherichia coli: A Crucial Biomolecule for Biotechnological and Healthcare Applications
Human Serum Albumin (HSA) is one of the most demanded therapeutic protein with immense biotechnological applications. The current source of HSA is human blood plasma. Blood is a limited and an unsafe source as it possesses the risk of contamination by various blood derived pathogens. This issue led to exploitation of various hosts with the aim to obtain an alternative source for the production of the rHSA. But, till now no host has been proven to be effective commercially for rHSA production because of their respective limitations. Thus, there exists an indispensable need to promote non-animal derived rHSA production. Of all the host systems, Escherichia coli is one of the most convenient hosts which has contributed in the production of more than 30% of the FDA approved recombinant pharmaceuticals. E. coli grows rapidly and its culture reaches high cell density using inexpensive and simple substrates. The fermentation batch turnaround number for E. coli culture is 300 per year, which is far greater than any of the host systems available. Therefore, E. coli derived recombinant products have more economical potential as fermentation processes are cheaper compared to the other expression hosts available. Despite of all the mentioned advantages, E. coli had not been successfully adopted as a host for rHSA production. The major bottleneck in exploiting E. coli as a host for rHSA production was aggregation i.e. majority of the expressed recombinant protein was forming inclusion bodies (more than 90% of the total expressed rHSA) in the E. coli cytosol. Recovery of functional rHSA form inclusion body is not preferred because it is tedious, time consuming, laborious and expensive. Because of this limitation, E. coli host system was neglected for rHSA production for last few decades. Considering the advantages of E. coli as a host, the present work has targeted E. coli as an alternate host for rHSA production through resolving the major issue of inclusion body formation associated with it. In the present study, we have developed a novel and innovative method for enhanced soluble and functional production of rHSA in E.coli (~60% of the total expressed rHSA in the soluble fraction) through modulation of the cellular growth, folding and environmental parameters, thereby leading to significantly improved and enhanced -expression levels as well as the functional and soluble proportion of the total expressed rHSA in the cytosolic fraction of the host. Therefore, in the present case we have filled in the gap in the literature, by exploiting the most well studied host system Escherichia coli which is of low cost, fast growing, scalable and ‘yet neglected’, for the enhancement of functional production of HSA- one of the most crucial biomolecule for clinical and biotechnological applications.
Biophysical Characterization of Archaeal Cyclophilin Like Chaperone Protein
Chaperones are proteins that help other proteins fold correctly, and are found in all domains of life i.e., prokaryotes, eukaryotes and archaea. Various comparative genomic studies have suggested that the archaeal protein folding machinery appears to be highly similar to that found in eukaryotes. In case of protein folding; slow rotation of peptide prolyl-imide bond is often the rate limiting step. Formation of the prolyl-imide bond during the folding of a protein requires the assistance of other proteins, termed as peptide prolyl cis-trans isomerases (PPIases). Cyclophilins constitute the class of peptide prolyl isomerases with a wide range of biological function like protein folding, signaling and chaperoning. Most of the cyclophilins exhibit PPIase enzymatic activity and play active role in substrate protein folding which classifies them as a category of molecular chaperones. Till date, there is not very much data available in the literature on archaeal cyclophilins. We aim to compare the structural and biochemical features of the cyclophilin protein from within the three domains to elucidate the features affecting their stability and enzyme activity. In the present study, we carry out in-silico analysis of the cyclophilin proteins to predict their conserved residues, sites under positive selection and compare these proteins to their bacterial and eukaryotic counterparts to predict functional divergence. We also aim to clone and express these proteins in heterologous system and study their biophysical characteristics in detail using techniques like CD and fluorescence spectroscopy. Overall we aim to understand the features contributing to the folding, stability and dynamics of the archaeal cyclophilin proteins.
Effect of Pioglitazone on Intracellular Na+ Homeostasis in Metabolic Syndrome-Induced Cardiomyopathy in Male Rats
Metabolic syndrome, is associated impaired blood glucose level, insulin resistance, dyslipidemia caused by abdominal obesity. Also, it is related with cardiovascular risk accumulation and cardiomyopathy. The hypothesis of this study was to examine the effect of thiazolidinediones such as pioglitazone which is widely used insulin-sensitizing agents that improve glycemic control, on intracellular Na+ homeostasis in metabolic syndrome-induced cardiomyopathy in male rats. Male Wistar-Albino rats were randomly divided into three groups, namely control (Con, n=7), metabolic syndrome (MetS, n=7) and pioglitazone treated metabolic syndrome group (MetS+PGZ, n=7). Metabolic syndrome was induced by providing drinking water that was 32% sucrose, for 18 weeks. All of the animals were exposed to a 12 h light – 12 h dark cycle. Abdominal obesity and glucose intolerance had measured as a marker of metabolic syndrome. Intracellular Na+ ([Na+]i) is an important modulator of excitation–contraction coupling in heart. [Na+]i at rest and [Na+]i during pacing with electrical field stimulation in 0.2 Hz, 0.8 Hz, 2.0 Hz stimulation frequency were recorded in cardiomyocytes. Also, Na+ channel current (INa) density and I-V curve were measured to understand [Na+]i homeostasis. In results, high sucrose intake, as well as the normal daily diet, significantly increased body mass and blood glucose level of the rats in the metabolic syndrome group as compared with the non-treated control group. In MetS+PZG group, the blood glucose level and body inclined to decrease to the Con group. There was a decrease in INa density and there was a shift both activation and inactivation curve of INa. Pioglitazone reversed the shift to the control side. Basal [Na+]i either MetS and Con group were not significantly different, but there was a significantly increase in [Na+]i in stimulated cardiomyocytes in MetS group. Furthermore, pioglitazone had not effect on basal [Na+]i but it reversed the increase in [Na+]i in stimulated cardiomyocytes to the that of Con group. Results of the present study suggest that pioglitazone has a significant effect on the Na+ homeostasis in the metabolic syndrome induced cardiomyopathy in rats. All animal procedures and experiments were approved by the Animal Ethics Committee of Ankara University Faculty of Medicine (2015-2-37).
A Cadaveric Study of Branching Pattern of Arch of Aorta and Its Clinical Significance in Nepalese Population
Background: The arch of aorta is a large artery that arches over the root of the left lung and connects the ascending aorta and descending aorta. It is situated in the superior mediastinum behind the manubrium sterni. It gives off three major branches i.e. brachiocephalic trunk, left common carotid artery and left subclavian artery arising from the superior surface of arch of aorta from right to left. Material and Methods: This was a descriptive study. It was carried out in 44 cadavers, obtained during dissections for undergraduates of Department of Anatomy, Chitwan Medical College, Bharatpur, Chitwan, between March 2015 to October 2016. Cadavers of both sexes were included in the present study. The arch of aorta was dissected and exposed according to the methods described by Romanes in Cunningham’s manual of practical anatomy. Results: Out of 44 dissected cadavers, 35 (79.54%) were male and 9 (20.46%) were female cadavers. The normal branching pattern of the arch of aorta was encountered in 28 (63.64%) cadavers and the remaining 16 (36.36%) cadavers showed variations in the branching pattern of arch of aorta. Two different types of variations on the branching pattern of arch of aorta were noted in the present study, in which 12 (27.27%) cadavers had common trunk of the Arch of Aorta. In 3 (5.00%) male cadavers, we found the origin of the Thyroid ima artery. This variation was noted in 1(1.66%) female cadaver. Conclusion: The present study carried out on adult human cadavers’ revealed wide variations in the branching pattern of the arch of ao rta. These variations are of clinical significance and also very useful for the anatomists, radiologists, anesthesiologists, surgeons for practice during angiography, instrumentation, supra-aortic thoracic, head and neck surgery.
Folding Pathway and Thermodynamic Stability of Monomeric GroEL
Chaperonin GroEL is a tetradecameric Escherichia coli protein having identical subunits of 57 kDa. The elucidation of thermodynamic parameters related to stability for the native GroEL is not feasible as it undergoes irreversible unfolding because of its large size (800kDa) and multimeric nature. Nevertheless, it is important to determine the thermodynamic stability parameters for the highly stable GroEL protein as it helps in folding and holding of many substrate proteins during many cellular stresses. Properly folded monomers work as building-block for the formation of native tetradecameric GroEL. Spontaneous refolding behavior of monomeric GroEL makes it suitable for protein-denaturant interactions and thermodynamic stability based studies. The urea mediated unfolding is a three state process which means there is the formation of one intermediate state along with native and unfolded states. The heat mediated denaturation is a two-state process. The unfolding process is reversible as observed by the spontaneous refolding of denatured protein in both urea and head mediated refolding processes. Analysis of folding/unfolding data provides a measure of various thermodynamic stability parameters for the monomeric GroEL. The proposed mechanism of unfolding of monomeric GroEL is a three state process which involves formation of one stable intermediate having folded apical domain and unfolded equatorial, intermediate domains. Research in progress is to demonstrate the importance of specific residues in stability and oligomerization of GroEL protein. Several mutant versions of GroEL are under investigation to resolve the above mentioned issue.
Study of Dermatoglyphics Pattern in Patient with Hypertension
Introduction: Dermatoglyphics is the science which deals with the study of dermal ridge configuration on the digits, palms and soles. It is grooved by ridges and forms variety of configurations. The aim of the study was to identify dermal ridge patterns on fingertip of hypertensive patients and in normal population and to compare patterns among them. Methods: The subjects of the study were 130 hypertensives and 130 non-hypertensives cases of Kathmandu Valley aged between 40 to 80 years. Case history was recorded after consent finger prints were taken. Different parameters as whorl, loop, arch and composite patterns were studied and analysed. Result: It revealed, increased whorl pattern in hypertensive. It showed 65.69% whorl, 29.23% loop and 5.07% arch patterns in right hand of hypertensive people. In control, it was found to be 34.46% whorl, 58.15% loop and 5.38% arch patterns respectively. Similarly in left hand 63.69% whorl, 32% loop and 4.30% arch in hypertensive group. In control group it was 60.15% as loop, 35.69% as whorl and 15% as arch. Discussion: Based on findings of the result, it was concluded that the whorl, loop and arch patterns observed as 65.69%, 29.23% and 5.07% respectively in hypertensive cases in right hand. Similarly in left hand, it was found to be 4.30% as arch, 32% as loop and 63.69% as whorl patterns, but in normotensive subjects these patterns were recorded as 36.43%, 58.15%, 5.38% in right hand and 35.69%, 60.15%, 4.15% in left hand as whorl, loop and arch respectively.
Feature Selection of Personal Authentication Based on EEG Signal for K-Means Cluster Analysis Using Silhouettes Score
Personal authentication based on electroencephalography (EEG) signals is one of the important field for the biometric technology. More and more researchers have used EEG signals as data source for biometric. However, there are some disadvantages for biometrics based on EEG signals. The proposed method employs entropy measures for feature extraction from EEG signals. Four type of entropies measures, sample entropy (SE), fuzzy entropy (FE), approximate entropy (AE) and spectral entropy (PE), were deployed as feature set. In a silhouettes calculation, the distance from each data point in a cluster to all another point within the same cluster and to all other data points in the closest cluster are determined. Thus silhouettes provide a measure of how well a data point was classified when it was assigned to a cluster and the separation between them. This feature renders silhouettes potentially well suited for assessing cluster quality in personal authentication methods. In this study, “silhouettes scores” was used for assessing the cluster quality of k-means clustering algorithm is well suited for comparing the performance of each EEG dataset. The main goals of this study are: (1) to represent each target as a tuple of multiple feature sets, (2) to assign a suitable measure to each feature set, (3) to combine different feature sets, (4) to determine the optimal feature weighting. Using precision/recall evaluations, the effectiveness of feature weighting in clustering was analyzed. EEG data from 22 subjects were collected. Results showed that: (1) It is possible to use fewer electrodes (3-4) for personal authentication. (2) There was the difference between each electrode for personal authentication (p< 0.01). (3) There is no significant difference for authentication performance among feature sets (except feature PE). Conclusion: The combination of k-means clustering algorithm and silhouette approach proved to be an accurate method for personal authentication based on EEG signals.
Automatic Differentiation of Ultrasonic Images of Cystic and Solid Breast Lesions
In most cases, typical cysts are easily recognized at ultrasonography. The specificity of this method for typical cysts reaches 98%, and it is usually considered as gold standard for typical cyst diagnosis. However, it is necessary to have all the following features to conclude the typical cyst: clear margin, the absence of internal echoes and dorsal acoustic enhancement. At the same time, not every breast cyst is typical. It is especially characteristic for protein-contained cysts that may have significant internal echoes. On the other hand, some solid lesions (predominantly malignant) may have cystic appearance and may be falsely accepted as cysts. Therefore we tried to develop the automatic method of cystic and solid breast lesions differentiation. Materials and methods. The input data were the ultrasonography digital images with the 256-gradations of gray color (Medison SA8000SE, Siemens X150, Esaote MyLab C). Identification of the lesion on these images was performed in two steps. On the first one, the region of interest (or contour of lesion) was searched and selected. Selection of such region is carried out using the sigmoid filter where the threshold is calculated according to the empirical distribution function of the image brightness and, if necessary, it was corrected according to the average brightness of the image points which have the highest gradient of brightness. At the second step, the identification of the selected region to one of lesion groups by its statistical characteristics of brightness distribution was made. The following characteristics were used: entropy, coefficients of the linear and polynomial regression, quantiles of different orders, an average gradient of brightness, etc. For determination of decisive criterion of belonging to one of lesion groups (cystic or solid) the training set of these characteristics of brightness distribution separately for benign and malignant lesions were received. To test our approach we used a set of 217 ultrasonic images of 107 cystic (including 53 atypical, difficult for bare eye differentiation) and 110 solid lesions. All lesions were cytologically and/or histologically confirmed. Visual identification was performed by trained specialist in breast ultrasonography. Results. Our system correctly distinguished all (107, 100%) typical cysts, 107 of 110 (97.3%) solid lesions and 50 of 53 (94.3%) atypical cysts. On the contrary, with the bare eye it was possible to identify correctly all (107, 100%) typical cysts, 96 of 110 (87.3%) solid lesions and 32 of 53 (60.4%) atypical cysts. Conclusion. Automatic approach significantly surpasses the visual assessment performed by trained specialist. The difference is especially large for atypical cysts and hypoechoic solid lesions with the clear margin. This data may have a clinical significance.
Nanotechnology-Based Treatment of Liver Cancer
We present method of Nanoparticle enhanced laser thermal ablation of HepG2 cells (Human hepatocellular liver carcinomacell line), using gold nanoparticles combuned with a specific growth factor and demonstrate its selective therapeutic efficacy usig ex vivo specimens. Ex vivo-perfused liver specimens were obtained from hepatocellular carcinoma patients similarly to the surgical technique of transplantation. Ab bound to GNPs was inoculated intra-arterially onto the resulting specimen and determined the specific delivery of the nano-bioconjugate into the malignant tissue by means of the capillary bed. The extent of necrosis was considerable following laser therapy and at the same time surrounding parenchyma was not seriously affected. The selective photothermal ablation of the malignant liver tissue was obtained after the selective accumulation of Ab bound to GNPs into tumor cells following ex-vivo intravascular perfusion. These unique results may represent a major step in liver cancer treatment using nanolocalized thermal ablation by laser heating.
Preparation and Characterizations of Hydroxyapatite-Sodium Alginate Nanocomposites for Biomedical Applications
Polymer-inorganic nanocomposites are presently impacting diverse areas, specifically in biomedical sciences. In this research, hydroxyapatite-sodium alginate has been prepared, and characterized, with emphasis on the influence of sodium alginate on its characteristics. In situ wet chemical precipitation method was used in the preparation. The prepared nanocomposite was characterized with Fourier Transform Infrared spectroscopy (FTIR), Scanning Electron Microscopy (SEM), with image analysis, and X-Ray Diffraction (XRD). The FTIR study shows peaks characteristics of hydroxyapatite and confirmed formation of the nanocomposite via chemical interaction between sodium alginate and hydroxyapatite. Image analysis shows the nanocomposites to be of irregular morphologies which did not show significant change with increasing sodium alginate addition, while particle size decreased with increase in sodium alginate addition (359.46 nm to 109.98 nm). From the XRD data, both the crystallite size and degree of crystallinity also decreased with increasing sodium alginate composition (32.36 nm to 9.47 nm and 72.87% to 1.82% respectively), while the specific surface area and microstrain increased with increasing sodium alginate composition (0.0041 to 0.0139 and 58.99 m²/g to 201.58 m²/g respectively). The results show that the formulation with 50%wt of sodium alginate (HASA-50%wt), possess exceptional characteristics for biomedical applications such as drug delivery.
Thermosensitive Polypeptide Hydrogel for Immunosuppressive Drug Delivery in Vascularized Composite Allotransplantation
Tacrolimus is an immunosuppressive agent which is usually applied to patients who received allotransplantation to reduce immune rejection. The strong hydrophobicity of tacrolimus makes it difficult to be applied in aqueous system. It has been studied to be encapsulated by micelles. However, the low efficiency of encapsulation in micelles is an obstacle for further application. Polypeptide thermosensitive hydrogel is considered as a great candidate for drug delivery because of several advantages, such as excellent biocompatibility and biodegradability, low concentration for gelation and sensitive response temperature change. In this study, we synthesized a thermo-responsive polypeptide hydrogel by copolymerizing poloxamer (PLX) and poly(ʟ-alanine) with ʟ-lysine segments at the both ends to form PLX-b-poly(ʟ-alanine-lysine) (P-Lys-Ala-PLX) copolymers. Poly(ʟ-alanine) is the hydrophobic chain of P-Lys-Ala-PLX copolymers which was designed to capture the hydrophobic agents. The synthesis was examined by 1H NMR and showed that P-Lys-Ala-PLX copolymers was successfully synthesized. P-Lys-Ala-PLX has a sol-gel behavior with low concentration of 3-7 wt% and has high degree of drug encapsulation. The experiment of drug release also showed that the tacrolimus released constantly over time. This study indicates that P-Lys-Ala-PLX is a high potential injectable drug carrier for biomedical application.
The Study of Dissolving Microneedle Patch for Androgenetic Alopecia
Microneedle patch is a painless transdermal drug delivery method; it could solve some problems in traditional drug delivery such as digestive system causing drug metabolism and subcutaneous injection causing some side effects. Coating drug on or loading drug in microneedle can carry active ingredient through stratum corneum, also can control dose well when microneedle patch is applied on localized topical area. We used hyaluronic acid to fabricate dissolvable microneedle patch and encapsulated minoxidil into microneedles. Minoxdil is a drug for an exterior use that can be used to treat Androgenetic alopecia, but related commercial products have some shortcomings, for example, propylene glycol which is used to soften stratum corneum cause skin allergic reaction, comparing chemical promotion, microneedle patch provide a physical way to make drugs through nature barrier of the skin. In this research, we designed a two-step process to fabricate microneedle patch, that can effectively reduce drug waste, and gentle production process could maintain drug activity well. We use Fluorescein Isothiocyanate Isomer I in microneedles to understand the distribution of the drug in the needle body, also do In vitro test on cadaver to make sure patch has enough mechanical strength to penetrate stratum corneum. In this study, we may determine that germinal MNs patch is a potential commodity.
Concept of Transforaminal Lumbar Interbody Fusion Cage Insertion Device
Transforaminal lumbar interbody fusion (TLIF) surgeries have nowadays became popular for treatment of degenerated spinal disorders. The interbody fusion technique like TLIF maintains load bearing capacity of the spine and a suitable disc height. Currently many techniques have been introduced to cure Spondylolisthesis. This surgery provides greater rehabilitation of degenerative spines. While performing this TLIF surgery existing methods use guideway, which is a troublesome surgery technique as the use of two separate instruments is required to perform this surgery. This paper presents a concept which eliminates the use of guideway. This concept also eliminates problems that occur like reverting the cage. The concept discussed in this paper also gives high accuracy while performing surgery.