Physical Sciences and Mathematics Commons^™

Discrimination Between Human And Non-Human Blood Using Raman Spectroscopy And A Self-Reference Algorithm For Forensic Purposes : Method Expansion And Validation, Harrison Dickler

Legacy Theses & Dissertations (2009 - 2024)

Determining whether the origin of a bloodstain is human or non-human is an important capability of a forensic investigation. In their pioneering work, Bian et al. (Journal of biomedical optics, 2017) introduced a self-reference peak algorithm for the analysis of bloodstain Raman spectra and demonstrated a great potential of this approach for differentiating human and nonhuman blood. However, this work only used three non-human species in the creation of their model. This study investigates the capability of a self-referencing peak algorithm to discriminate between human and 18 non-human species based on the Raman spectra of blood samples. The intensity ratios …

Hemoglobin-Modified Core–Shell Fe3o4@Au Nanostructures For The Electrochemical Detection Of Acrylamide, Endang Saepudin, Tri Yuliani, Mochammad Arfin Fardiansyah Nasution, Munawar Khalil, Jong Wook Hong, Tribidasari Anggraningrum Ivandini

Makara Journal of Science

In this study, electrochemical detection of acrylamide using hemoglobin (Hb)-modified core–shell Fe₃O₄@Au nanostructures was conducted. Fe₃O₄ nanoparticles (~4.9 nm) and core–shell Fe₃O₄@Au (5.0–6.4 nm) nanostructures were successfully synthesized by the thermal decomposition method. Electrochemical investigation revealed that the optimum amount of Hb of 2 mg/mL could be immobilized in 0.1 M acetate buffer solution (pH = 6). Moreover, the detection of acrylamide using Fe₃O₄@Au/Hb was evaluated by the cyclic voltammetry technique. A linear calibration curve (R² = 0.98) in the concentration range of …

Oxygen Binding Thermodynamics Of Human Hemoglobin In The Red Blood Cell, Kyle K. Hill

Department of Chemistry: Dissertations, Theses, and Student Research

We report for the first time the binding constants and Hill numbers for oxygen in the red blood cell under physiological conditions. When compared to our results for hemoglobin in solution, our results show conclusively that hemoglobin binds oxygen more tightly and with lower co-operativity when packed in the red blood cell. At 18°C, these differences are striking: the respective half-saturation values are 15.57 µM (in red blood cells) and 18.83 µM (in solution), with corresponding Hill numbers of 2.475 (in red blood cells) and 2.949 (in solution). The optical complications that arise from high turbidity of red blood cell …

Electrochemical Sensor Based On Magnetic Electrode Modified With Magnetic Molecularly Imprinted Nanoparticles Immobilized Hemoglobin For Determination Of Hydrogen Peroxide, Yang Yuan, Jia-Xin Wang, Yu-Hua Cao

Journal of Electrochemistry

In this work, the surface-imprinted technique was used to prepare magnetic hemoglobin (Hb) imprinted nanoparticles, using Fe₃O₄@SiO₂ NPs as the carrier, Hb as the template molecule, and tetraethyl orthosilicate (TEOS) as the imprinted polymer monomer. The nanoparticles had a core-shell structure, with magnetic Fe₃O₄ NPs as the core and Hb imprinted polymers as the shell. Therefore, Hb could be concentrated and fixed on the surface of the magnetic imprinted nanoparticles (MMIPs NPs). Furthermore, MMIPs NPs were immobilized with chitosan (CS) on the surface of a magnetic electrode to constitute Hb enzyme-like biosensor …

Quantification Of Hemoglobin In Human Blood To Differentiate Individuals From One Another, Audrey Jeanne Auleley

Legacy Theses & Dissertations (2009 - 2024)

Blood is one of the principal sources of evidence in crime scene investigations. The level of hemoglobin in human blood is known to hardly fluctuate over time in a healthy adult patient. Using a method like the sodium lauryl sulfate (SLS) method for hemoglobin level determination is beneficial because SLS converts any other derivates of hemoglobin into methemoglobin. The amount of hemoglobin would be determined using the linear correlation between the absorbance of methemoglobin and its amount in blood samples. Those curves differ significantly from sample to sample. Therefore, a differentiation can be observed between the blood samples of different …

Effects Of Age, Mass, And Terrestrial Duration On Oxygen Store Development In Phocids And Otariids, Rachael M. Stevenson

HCNSO Student Capstones

Diving ontogeny among pinniped species was explored to determine how oxygen store development was affected by age, mass, and terrestrial duration between Phocidae and Otariidae. Distinct physiological and post-natal developmental differences exist between the two families which contribute to oxygen store and diving development. Blood oxygen data for six phocids and otariids and muscle oxygen data for two phocids and otariids were obtained. Body mass significantly (p

Erythrocytes And Vascular Function: Oxygen And Nitric Oxide, Christine C. Helms, M. T. Gladwin, Daniel B. Kim-Shapiro

Physics Faculty Publications

Erythrocytes regulate vascular function through the modulation of oxygen delivery and the scavenging and generation of nitric oxide (NO). First, hemoglobin inside the red blood cell binds oxygen in the lungs and delivers it to tissues throughout the body in an allosterically regulated process, modulated by oxygen, carbon dioxide and proton concentrations. The vasculature responds to low oxygen tensions through vasodilation, further recruiting blood flow and oxygen carrying erythrocytes. Research has shown multiple mechanisms are at play in this classical hypoxic vasodilatory response, with a potential role of red cell derived vasodilatory molecules, such as nitrite derived nitric oxide and …

Effects Of Mutations On The Molecular Dynamics Of Oxygen Escape From The Dimeric Hemoglobin Of Scapharca Inaequivalvis, Kevin Trujillo, Tasso Papagiannopoulos, Kenneth W. Olsen

Ken Olsen

Like many hemoglobins, the structure of the dimeric hemoglobin from the clam Scapharca inaequivalvis is a “closed bottle” since there is no direct tunnel from the oxygen binding site on the heme to the solvent. The proximal histidine faces the dimer interface, which consists of the E and F helicies. This is significantly different from tetrameric vertebrate hemoglobins and brings the heme groups near the subunit interface. The subunit interface is also characterized by an immobile, hydrogen-bonded network of water molecules. Although there is data which is consistent with the histidine gate pathway for ligand escape, these aspects of the …

Synthesis Of Multifunctional Polyacrylates And A Binding Group To Hemoglobin For The Treatment Of Traumatic Brain Injuries, Marina Michaud

Honors College Theses

Hemoglobin based oxygen carriers (HBOCs) hold promise as an effective emergency treatment of severe traumatic brain injuries (TBI). In the latest generation of HBOCs, polynitroxyl-pegylated hemoglobin (PNPH), cell-free hemoglobin is modified with TEMPO and PEG which reduce the toxicities associated with earlier generations of HBOCs. In our efforts to optimize the economic and therapeutic impacts of PNPH’s we have synthesized polydimethylaminoethyl methacrylate (poly-DMAEMA) under controlled living conditions via reverse addition-fragmentation chain transfer (RAFT) polymerization. The poly-DMAEMA was then successfully functionalized via quaternization of its NMe₂ groups using chloroacetate derivatives of the TEMPO and PEG. This process was quantitative and …

Recent Insights Into Nitrite Signaling Processes In Blood, Christine C. Helms, Xiaohua Liu, Daniel B. Kim-Shapiro

Physics Faculty Publications

Nitrite was once thought to be inert in human physiology. However, research over the past few decades has established a link between nitrite and the production of nitric oxide (NO) that is potentiated under hypoxic and acidic conditions. Under this new role nitrite acts as a storage pool for bioavailable NO. The NO so produced is likely to play important roles in decreasing platelet activation, contributing to hypoxic vasodilation and minimizing blood-cell adhesion to endothelial cells. Researchers have proposed multiple mechanisms for nitrite reduction in the blood. However, NO production in blood must somehow overcome rapid scavenging by hemoglobin in …

Binding Of Oxygen To Human Hemoglobin Within The Erythrocyte Using Icam Spectrophotometry, Kyle K. Hill

Department of Chemistry: Dissertations, Theses, and Student Research

Many of the spectrophotometric techniques used to determine the properties of intracellular human hemoglobin cannot be utilized due to the turbidity of erythrocyte suspensions. An Integrating Cavity Absorption Meter, or ICAM, allows for absorption measurements of strongly scattering samples in the visible-light region of the spectrum. The spectrum of oxygenated hemoglobin within erythrocytes is significantly different from the absorption spectrum of oxygenated hemoglobin in solution. Studies of the oxygen binding to hemoglobin in erythrocytes allowed the four sequential binding constants (Adair constants) to be determined and compared with those of hemoglobin in solution. The Adair constants for hemoglobin in solution …

Utilizing In Silico And/Or Native Esi Approaches To Provide New Insights On Haptoglobin/Globin And Haptoglobin/Receptor Interactions, Ololade Fatunmbi

Doctoral Dissertations

Haptoglobin (Hp), an acute phase protein, binds free hemoglobin (Hb) dimers in one of the strongest non-covalent interactions known in biology. This interaction protects Hb from causing potentially severe oxidative damage and limiting nitric oxide bioavailability. Once Hb/Hp complexes are formed, they proceed to bind CD163, a cell surface receptor on macrophages leading to complex internalization and catabolism. Myoglobin, (Mb) a monomeric protein, that is normally found in the muscle but can be released into the blood in high concentrations during myocardial injury, is homologous to Hb and shares many conserved Hb/Hp interface residues. Both monomeric Hb and Mb species …

Hydrogen Exchange Mass Spectrometry For Studying Protein-Ligand Interactions, Modupeola A. Sowole

Electronic Thesis and Dissertation Repository

Hydrogen deuterium exchange (HDX) coupled with mass spectrometry is widely used for probing protein structure and dynamics. Protein-ligand interactions usually induce a reduction in the measured HDX rates an effect that may be ascribed to stabilization of the protein structure. This work aims to improve the general understanding of the changes in HDX patterns associated with ligand binding.

We initially applied HDX for studying differences between oxy-hemoglobin (Oxy-Hb) and aquomet-hemoglobin (Chapter 2). The results show that the α and β subunits respond differently to the oxy to aquomet transition with the heme binding pocket being destabilized in both cases. The …

Structural Studies Of A Circularly Permuted Human Hemoglobin Containing Low O₂-Affinity Mutations, Rachel Hubbard

Scholars Week

Our research is focused on the production of a hemoglobin-based oxygen carrier (HBOC) which can be used as a therapeutic in the event of acute blood loss. The administration of cell-free hemoglobin is associated with severe adverse effects due to dissociation of the tetrameric α₂β₂ complex into αβ heterodimers. Our approach to designing an effective HBOC is based on a recombinant circularly permuted human hemoglobin in which all of the subunits are linked in a single-chain fashion. This design would prevent the dissociation of the tetramer and allow for the biosynthesis of polymeric hemoglobins of defined mass. Preliminary ligand binding …

Effects Of Mutations On The Molecular Dynamics Of Oxygen Escape From The Dimeric Hemoglobin Of Scapharca Inaequivalvis, Kevin Trujillo, Tasso Papagiannopoulos, Kenneth W. Olsen

Chemistry: Faculty Publications and Other Works

Like many hemoglobins, the structure of the dimeric hemoglobin from the clam Scapharca inaequivalvis is a “closed bottle” since there is no direct tunnel from the oxygen binding site on the heme to the solvent. The proximal histidine faces the dimer interface, which consists of the E and F helicies. This is significantly different from tetrameric vertebrate hemoglobins and brings the heme groups near the subunit interface. The subunit interface is also characterized by an immobile, hydrogen-bonded network of water molecules. Although there is data which is consistent with the histidine gate pathway for ligand escape, these aspects of the …

Preparation Of Surface Molecularly Imprinted Magnetic Nanoparticles For Hemoglobin Sensing, Chun-Fang Huang, Gui-Hong Yao, Jian-Ding Qiu

Journal of Electrochemistry

Surface molecularly imprinted magnetic nanoparticles (NPs) were prepared by employing dopamine (DA) as a functional monomer, hemoglobin (Hb) as a template, H₂PtCl₆ as an oxidant triggered DA polymerization on the surface of the Fe₃O₄ NPs. The Hb and the formed platinum nanoparticles (PtNPs) were embedded into the polydopamine (PDA). After removal of the Hb, the surface molecularly imprinted magnetic nanoparticles (imprinted Fe₃O₄/PDA-PtNPs) were formed, and then simply immobilized on the magnetic glassy carbon electrode (imprinted Fe₃O₄/PDA-PtNPs/MGCE) for Hb sensing. The imprinted Fe₃O₄/PDA-PtNPs …

Polyethylene Glycol Containing Rompolymers For The Modification Of Neuro-Protective Hemoglobin, Andrea L. Mccollum

Honors College Theses

Polyethylene glycol (PEG) has shown the ability to improve compatibility when used in combination with cell-free hemoglobin in the treatment of traumatic brain injuries. It has been demonstrated that the covalently bonded PEG increases the hydrodynamic radius of the hemoglobin and hence generates a physical barrier while slowing down the oxygen delivery of the strongly oxidative hemoglobin. In this context, I have been working on the development of synthetic pathways to incorporate PEG into monomers and polymers through both direct modification of (7-oxa)norbornene derivatives and post polymerization modification. Starting from the (7-oxa)norbornene anhydride derivatives, we have developed pathways to cationic …

Mechanisms Of Hemolysis-Associated Platelet Activation, Christine C. Helms, M. Marvel, W. Zhao, M. Stahle, R. Vest, G. J. Kato, J. S. Lee, G. Christ, M. T. Gladwin, R. R. Hantgan, D. B. Kim-Shapiro

Physics Faculty Publications

Background

Intravascular hemolysis occurs after blood transfusion, in hemolytic anemias, and in other conditions, and is associated with hypercoagulable states. Hemolysis has been shown to potently activate platelets in vitro and in vivo, and several mechanisms have been suggested to account for this, including: (i) direct activation by hemoglobin (Hb); (ii) increase in reactive oxygen species (ROS); (iii) scavenging of nitric oxide (NO) by released Hb; and (iv) release of intraerythrocytic ADP.

Objective

To elucidate the mechanism of hemolysis-mediated platelet activation.

Methods

We used flow cytometry to detect PAC-1 binding to activated platelets for in vitro experiments, and a …

Hemoglobin-Mediated Nitric Oxide Signaling, Christine C. Helms, D. B. Kim-Shapiro

Physics Faculty Publications

The rate that hemoglobin reacts with nitric oxide (NO) is limited by how fast NO can diffuse into the heme pocket. The reaction is as fast as any ligand/protein reaction can be and the result, when hemoglobin is in its oxygenated form, is formation of nitrate in what is known as the dioxygenation reaction. As nitrate, at the concentrations made through the deoxygenation reaction, is biologically inert, the only role hemoglobin was once thought to play in NO signaling was to inhibit it. However, there are now several mechanisms that have been discovered by which hemoglobin may preserve, control, and …

Protein Structure And Interactions Studied By Electrospray Mass Spectrometry, Jiangjiang Liu

Electronic Thesis and Dissertation Repository

Since the emergence of electrospray ionization (ESI) mass spectrometry (MS) as a tool for protein structural studies, this area has experienced tremendous growth. ESI-MS is highly sensitive, and it allows the analysis of biological systems ranging in size from a few atoms to large multi-protein complexes. This work aims to solve questions in protein structural biology by using ESI-MS in conjunction with other techniques.

We initially apply ESI-MS for studying the monomeric protein cytochrome c (Chapter 2). The physical reasons underlying the irreversible thermal denaturation of this protein remain controversial. By utilizing deconvoluted charge state distributions, oxidative modifications were found …

A Hemoglobin-Based Multifunctional Therapeutic: Polynitroxylated Pegylated Hemoglobin, Carleton C. J. Hsia, Li Ma

Department of Physics and Astronomy Faculty Publications

Polynitroxylated pegylated hemoglobin (PNPH) as a multifunctional therapeutic takes advantage of the ability of hemoglobin (Hb) to transport oxygen, the antioxidative stress activities from the redox coupling of nitroxide and heme iron, and the hypercolloid properties of pegylation. The published preclinical data demonstrating that PNPH acts as a neurovascular protective multifunctional therapeutic in an animal model simulating prehospital resuscitation of traumatic brain injury (TBI) with hemorrhagic shock (HS) are reviewed. Preliminary results on the potential utility of PNPH for neurovascular protection in thrombolytic stroke therapy and for correction of vascular dysfunction through transfusion in sickle-cell disease (SCD) are also discussed. …

Determination Of Lidocaine Based On Electrocatalysis Of A Chemically Modified Electrode, Gamze Tan, Gülçi̇n Bolat, Mehmet Ali̇ Onur, Serdar Abaci

Turkish Journal of Chemistry

The potential application of biochemically modified electrodes was investigated for construction of a drug biosensor. The electrocatalytic response of lidocaine was examined through modified electrodes using the electron transfer ability of hemoglobin (Hb) immobilized on gold film modified with a self-assembled monolayer (SAM) of 3-mercaptopropionic acid (MPA). According to the results of voltammetric studies, Au/MPA/Hb electrodes exhibited high sensitivity and good electrocatalytic activity for the reduction of lidocaine, motivating us to construct drug biosensors. With this study, a new electroanalytical procedure is proposed for the determination of lidocaine level in medical applications or therapeutic formulations containing lidocaine as the active …

Influence Of Electric Field On The Adsorption And Bioactivity Of Hemoglobin On A Macroporous Gold Electrode, Cui-Hong Wang, Yi-Bing Kong, Xing-Hua Xia

Journal of Electrochemistry

The adsorption and bioactivity of proteins at interfaces have been widely studied due to their important role in the construction of biosensors,bioelectronics and biofuel cells.Interfacial electric field is one of the important factors which could affect the adsorption and bioactivity of proteins at materials surfaces.It could dramatically change the adsorption density,molecular conformation and orientation at material surfaces.In this paper,the influence of interfacial electric field on the adsorption kinetics and bioactivity of hemoglobin(Hb) on a three-dimensional(3D) macroporous gold electrode surface has been studied using electrochemical methods and infrared spectroscopy.It was found that the interfacial electric field created excess surface charge which …

From Powder To Solution: Hydration Dependence Of Human Hemoglobin Dynamics Correlated To Body Temperature, A. M. Stadler, I. Digel, J. P. Embs, T. Unruh, Moeava Tehei, G. Zaccai, G. Büldt, G. M. Artmann

Faculty of Science - Papers (Archive)

A transition in hemoglobin (Hb), involving partial unfolding and aggregation, has been shown previously by various biophysical methods. The correlation between the transition temperature and body temperature for Hb from different species, suggested that it might be significant for biological function. In order to focus on such biologically relevant human Hb dynamics, we studied the protein internal picosecond motions as a response to hydration, by elastic and quasielastic neutron scattering. Rates of fast diffusive motions were found to be significantly enhanced with increasing hydration from fully hydrated powder to concentrated Hb solution. In concentrated protein solution, the data revealed that …

Direct Electrochemistry Of Hemoglobin Immobilized On The Electrode Modified With Ionic Liquid And Kgm, Liu Yang, Xia-Qin Wu, Wen-Jing Hou, Lin-Fei Gu, Rong Wang, Xiao-Ming Guo, Zong-Rang Zhang

Journal of Electrochemistry

Hemoglobin(Hb) has been immobilized on a glassy carbon electrode(GCE) surface with konjac glucomannan(KGM) and room temperature ionic liquid(RTIL).And which electrochemical behaviors have been investigated by cyclic voltammetry.The results shown that the entrapped Hemoglobin undergoes fast direct electron transfer reactions with formal potential of-0.38V(vs.SCE) and the E0' is linearly dependent on solution pH with a slope value of 51 mV/pH,due to the electron transfer of HbFe3+/HbFe2+ redox couple companied with a proton transfer.The electrocatalytic reduction of oxygen was also explored at the Hb-KGM/RTIL/Nafion/ GC electrode.

Mechanisms Of Hemoglobin Adaptation To High Altitude Hypoxia, Jay F. Storz, Hideaki Moriyama

Hideaki Moriyama Publications

Evidence from a number of vertebrate taxa suggests that modifications of hemoglobin (Hb) function may often play a key role in mediating an adaptive response to high altitude hypoxia. The respiratory functions of Hb are a product of the protein’s intrinsic O₂-binding affinity and its interactions with allosteric effectors such as protons, chloride ions, CO₂, and organic phosphates. Here we review several case studies involving high altitude vertebrates where it has been possible to identify specific mechanisms of Hb adaptation to hypoxia. In addition to comparative studies of Hbs from diverse animal species, functional studies of …

Mechanisms Of Hemoglobin Adaptation To High Altitude Hypoxia, Jay F. Storz, Hideaki Moriyama

Hideaki Moriyama Publications

Evidence from a number of vertebrate taxa suggests that modifications of hemoglobin (Hb) function may often play a key role in mediating an adaptive response to high altitude hypoxia. The respiratory functions of Hb are a product of the protein’s intrinsic O₂-binding affinity and its interactions with allosteric effectors such as protons, chloride ions, CO₂, and organic phosphates. Here we review several case studies involving high altitude vertebrates where it has been possible to identify specific mechanisms of Hb adaptation to hypoxia. In addition to comparative studies of Hbs from diverse animal species, functional studies of …

Voltammetric Behavior And Electrocatalytic Properties Of Hemoglobin At Glassy Carbon Electrode Modified With Lauric Acid, Xin-Rui Miao, Xu-Hong Zhang, Ying Xie

Journal of Electrochemistry

The electrochemical behavior of hemoglobin at glassy carbon(GC)electrode modified with Lauric Acid was studied.In the potential of +0.6～-0.7 V(vs.Ag/AgCl),an irreversible cathodic peak was observed at-0.3 V at the scan rate of 20 mV·s-1.In the medium of 0.02 mol·L-1 KH2PO4-Na2HPO4(pH 7.0),ip increased with the increasing concentration of Hb and the value of pH.The peak currents were proportional to the concentrations of hemoglobin in the range of 1.00×10-8～5.00×10-9 mol·L-1 and 1.92×10-6～2.06×10-7 mol·L-1.The data was analyzed in details by simulating in order to obtain more information and reaction mechanism of Hb on modified electrode.The electrode can be used as an approach for determining …

Vascular Morphometry Of The Retina In Antarctic Fishes Is Dependent Upon The Level Of Hemoglobin In Circulation, Jody M. Wujcik

Electronic Theses and Dissertations

Antarctic notothenioids express the circulating oxygen-binding protein hemoglobin (Hb) over a broad range of blood concentrations. White-blooded icefishes (Suborder: Notothenioidei, Family: Channichthyidae) are the only known adult vertebrates to lack Hb completely. In addition to its role in oxygen transport, Hb is the primary reactant in degradation of nitric oxide (NO). Thus, NO should be degraded at a slower rate in Hb-lacking icefishes than in Hb-expressing notothenioids, leading to higher steady-state levels of NO in the former group. Increased levels of NO should stimulate upregulation of angiogenesis, the growth and proliferation of new blood vessels from existing vasculature. Based upon …

Electrochemical Behaviours Of Hemoglobin-Ddab-Modified Powder Microelectrode And The Application Of The Powder Microelectrode In Determination Of Nitric Oxide, Zhi-Mou Guo, Huan Liu, Jian Chen, Wen-Ting Zhang

Journal of Electrochemistry

The electrochemical behaviours of DDAB-Hb-PME were studied in this paper. The (apparent) surface concentration of Hb was enhanced greatly by using the powder microelectrode (technique). NO reacts with Hb and absorbs on the surface of the modified carbon powder. Therefore, the output of the electrode was enhanced greatly by employing PME with higher reaction surface (area). The sensitivity of DDAB-Hb-PME is 3.1 mA/μmol·L~(-1)·cm~(2), which is much larger than the values obtained with the traditional electrodes. The detection limit is 9 nmol·L~(-1) and the linear response range is 9～100 nmol·L~(-1).