Body Fluids

Hybridomas, Genetic Engineering of

Michael Butler , in Encyclopedia of Physical Science and Technology (Third Edition), 2003

I Introduction: The Nature of Antibodies

Antibodies are glycoproteins found in body fluids including blood, milk, and mucous secretions and serve an essential role in the immune system that protects animals from infection or the cytotoxic effects of foreign compounds. Antibodies will bind with high affinity to an invasive molecule. Normally the binding is to only part of a large molecule (the epitope) and so there may be many different antibodies for a particular compound. Antibodies have become essential tools for biological research because of their very specific recognition and affinity for one compound (the antigen). This has not only led to the use of antibodies in the recognition of specific cellular components but also to the development of routine diagnostic medical tests. More recently antibodies have been used as therapeutic agents for the treatment of human disease.

Each B-lymphocyte is capable of producing one type of antibody in response to a particular antigen which interacts with a cell surface receptor. Stimulation by an antigen causes growth and an expansion of the cell population capable of producing the corresponding antibody. The variety of antibodies present in any animal reflects the population of B-lymphocytes which have been stimulated by previous exposure to a range of antigens.

Antibodies are found in a specific protein fraction of blood called the gamma-globulin or the immunoglobulin fraction. They are synthesized by a subset of white blood cells—the B-lymphocytes. The molecular structures of the five major classes (isotypes) of immunoglobulins (IgM, IgD, IgG, IgE, and IgA) are shown in Fig. 1. The basic structural arrangement of two heavy associated with two light chains is similar for all the isotypes. However, each isotype is distinguished by different heavy chain structures which are of varying length, number of domains, and glycan structures. The glycans are indicated by the fork structures (∐). It is also to be noted that the IgM configuration consists of five basic structures linked as a pentamer.

FIGURE 1. Structures of immunoglobulin isotypes.

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Physical Medicine and Rehabilitation

J. Watkins , in Comprehensive Biomedical Physics, 2014

10.01.2.2 Intercellular Exchange

In multicellular organisms, the cells rely on the circulating body fluids such as blood to supply them with nutrients, oxygen, and other substances and to carry away waste products such as carbon dioxide. This involves exchange of nutrients, gases, and other substances between the vessels of the circulating body fluids and cells adjacent to the vessels, and between cells adjacent to each other. Intercellular exchange ensures that all cells are supplied with nutrients, gases, and other substances and can excrete waste products, even if the cells do not receive a direct supply of the circulating body fluids.

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Environmental Geochemistry

G.S. Plumlee , T.L. Ziegler , in Treatise on Geochemistry, 2007

9.07.5.8 Summary—Body Fluids from a Geochemical Perspective

As summarized in Table 4 and Figures 4–7 , various human body fluids with which Earth materials may come into contact exhibit a wide range of compositions, including their pH and Eh, electrolyte content, and concentrations of organic species such as amino acids, organic acids, and proteins. This compositional variability indicates that any given mineral or Earth material, and its contained metals, may potentially behave quite differently from a geochemical perspective depending upon the exposure pathway, the resulting body fluid(s) it encounters, and how it may modify body-fluid chemistry.

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Forensic Science, Applications of Mass Spectrometry*

Rodger L. Foltz , ... Dennis J. Crouch , in Encyclopedia of Spectroscopy and Spectrometry (Second Edition), 1999

Postmortem Toxicology

Postmortem forensic toxicology involves the analysis of drugs and poisons in body fluids and tissues collected from victims that have died under suspicious circumstances. Postmortem forensic toxicology is a key part of a death investigation in that it can provide information addressing the following questions and issues: (1) What drugs or toxins are present in the body? (2) Are the concentrations of the drugs or toxins in the lethal range? (3) Could drugs have caused impairment resulting in a fatal accident?

Currently in the United States there are no federally mandated regulations governing postmortem forensic toxicology. However, several forensic toxicology accreditation programmes have been established, and the Society of Forensic Toxicology and the American Academy of Forensic Sciences have published guidelines for forensic toxicology laboratory procedures and performance.

Unlike workplace drug testing, in which urine samples are analysed for a relatively small number of drugs and drug metabolites, postmortem forensic toxicology often involves analysis for a wide variety of drugs and poisons in many different specimens, including gastric contents, vitreous humor, blood, liver, spleen, muscle and brain tissue. Analysis of blood is particularly important because drug concentrations in blood that are high enough to cause death are known for most drugs. However, drug concentrations in blood are generally lower than in urine or in tissues.

Before extraction and analysis, tissues must be homogenized. Water or buffer solutions such as 0.1   M sodium phosphate, pH 6, are added to the tissue sample prior to homogenization. It is important to record the weight of the tissue and the volume of the fluid in which the tissue is homogenized. This information will be used to express the amount of drug per gram of tissue.

Extraction efficiency and sample cleanliness can be a particular challenge in the analysis of postmortem samples because the samples are often collected from decomposed bodies. Products from decomposition can reduce the extraction efficiency and can produce interfering peaks. Also, tissue homogenates contain lipid material that must be separated from the drug analyte prior to GC-MS analysis. Basic drugs can be efficiently separated from lipid material by back-extraction. In this procedure the drug is extracted from the tissue homogenate into a water-immiscible organic solvent and then back-extracted into a dilute acid solution while the neutral lipid material remains in the organic solvent. The dilute acid solution is then made basic and the drug is re-extracted into an organic solvent.

Another cleanup procedure is to reconstitute the initial extract residue in a mixture of hexane and ethanol–water (80:20 v/v). Most drugs will partition into the ethanol–water layer and lipid compounds will partition into the hexane layer. Precipitation of proteins from the tissue homogenates prior to extraction is also helpful for improving cleanliness of the extract and increasing the extraction efficiency. Reagents that precipitate proteins include acetonitrile, methanol and trichloroacetic acid. Digestion of tissue samples with proteolytic enzymes such as subilisin A can also enhance the recovery of drugs in the extraction procedure.

Following extraction, the analyte is derivatized, if necessary, and analysed by GC-MS. As discussed previously, electron ionization continues to be favoured. However, use of chemical ionization is increasing because it often provides better sensitivity. Also, a chemical ionization mass spectrum will frequently provide valuable complementary information. For example, Figure 2 shows electron ionization and chemical ionization mass spectra for two similar drugs, amitriptyline and cyclobenzaprine. The electron ionization mass spectra for the two drugs are virtually identical owing to the prominence of the common fragment ion at m/z 58. The positive-ion chemical ionization mass spectra show protonated molecule ion peaks that differ by two daltons, thereby permitting identification of the two drugs.

Figure 2. Comparison of the electron ionization and positive-ion chemical ionization mass spectra of cyclobenzaprine and amitriptyline.

Of course, the relevant questions in a death investigation cannot be answered by mass spectrometric analysis and other analytical data alone. Final interpretation should always include careful consideration of all the information relating to the case.

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Mass Spectrometry in Forensic Science

Jan Schuberth , in Encyclopedia of Physical Science and Technology (Third Edition), 2003

II.A Toxicology

The search for drug substances, pesticides, poisons, and their metabolites in body fluids from living persons or in postmortem organs presents an important and difficult task for the chemist. In addition to the fact that the forensic scientist most often does not know what intoxicant to look for, the main reason for using MS to begin with is the large number of possible toxic substances. In toxicology work at the Poison Center in Munich, as many as 8000 different substances have, in fact, been reported in 40,000 investigated objects.

Figure 9 shows an outline of the usual MS approach for searching a biological sample taken from a human for alien compounds with pharmacological effects. The example selected is a real-life incident of reckless driving by a motorist apprehended by the police on the suspicion of being under the influence of some drug or drugs. An extract of the blood sample was injected into a gas chromatograph/mass spectrometer focused on a broad range of mass fragments. The mass chromatogram at A in the figure is made up of the total ion current (the sum of all fragments recorded) and showed no clear peaks indicative of any drug substances. To raise the signal-to-noise ratio, the total ion current was reconstructed with the sum of the m/z 91 and 92, and then a peak appeared on the new mass chromatogram at B.

FIGURE 9. Search of blood sample for toxics. The mass chromatogram at A shows the total ion current (the sum of all fragments recorded) and the reconstructed mass chromatogram at B shows the ions with the sum of m/z 91 and 92. The mass spectra at C–F depict the library search for identifying the peak at B. The unknown analyte's mass spectrum is, after background subtraction, displayed at C. The three hottest candidates in the library along with their names and CAS (Chemical Abstracts Service) numbers are shown at D–F. At G are shown the chemical formula of the first ranked candidate and the value for how well the mass spectrum of the candidate fits with that of the analyte and vice versa. A value of 1000 indicates identical mass spectra; zero, no fragments in common.

In the next step of the analytical process, the substance generating the peak at B was to be identified, which was achieved by comparing the mass spectrum of the analyte at C with mass spectra in an on-line library. Out of ten candidates picked by the program, three possible ones are shown at D, E, and F. Even though the mass spectrum of the analyte best fitted that of methylbenzene, it also matched nearly as well the mass spectra of the two other candidates. In addition to the recorded fragments at m/z 91 and 92, the final identification of the analyte was based on the fact that the retention time for the analyte was the same as that for methylbenzene. To hold up to legal scrutiny, proof of the analyte identity generally must indicate that at least two fragments and the retention time are the same as for the suggested substance. In the example here, the motorist suspected of being under the influence of drugs was probably a "sniffer," who had inhaled paint thinner or some other solvent containing toluene (methylbenzene) before driving his car.

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WIND-INDUCED VIBRATIONS

T. Kijewski , ... A. Kareem , in Encyclopedia of Vibration, 2001

Aeroelastic Effects

The determination of wind-induced loads and response discussed previously did not account for aeroelastic effects, which can sometimes have significant contributions to the structural response. Response deformations can alter the aerodynamic forces, thus setting up an interaction between the elastic response and aerodynamic forces commonly referred to as aeroelasticity. Aeroelastic contributions to the overall aerodnamic loading are distinguished from other unsteady loads by recognizing that aeroelastic loads vanish when there is no structural motion. Different types of aeroelastic effects are commonly distinguished from each other. They include vortex-induced vibration, galloping, flutter, and aerodynamic damping.

As alluded to earlier, aerodynamically bluff cross-sections shed vortices at a frequency governed by the nondimensional Strouhal number, St:

[19] S t = f s b U ¯

where f s is the shedding frequency (in Hz). The shedding of vortices generates a periodic variation in the pressure over the surface of the structure. When the frequency of this variation approaches one of the natural frequencies of a structure, vortex-induced vibration can occur. The magnitudes of these vibrations are governed both by the structure's inherent damping characteristics and by the mass ratio between the structure and the fluid it displaces. These two effects are often combined in the Scruton number defined as:

[20] S c = 4 π ζ m ρ b 2

where m is the mass per unit length of the structure.

Vortex-induced vibration is more complex than a mere resonant forcing problem. Nonlinear interaction between the body motion and its wake results in the 'locking in' of the wake to the body's oscillation frequency over a larger velocity range than would be predicted using the Strouhal number. Vortex-induced vibration, therefore, occurs over a range of velocities that increases as the structural damping decreases.

Galloping occurs for structures of certain cross-sections at frequencies below those of vortex-induced vibration. One widely known example of galloping is the large acrosswind amplitudes exhibited by power lines when freezing rain has resulted in a change of their cross-section. Analytically, galloping is considered a 'quasisteady' phenomenon because knowledge of the static aerodynamic coefficients of a given structure (i.e., mean lift and drag forces on a stationary model) allows quite reliable prediction of galloping behavior.

Stability of aeroelastic interactions is of crucial importance. The attenuation of structural oscillations by both structural and aerodynamic damping characterizes stable flow-structure interactions. In an unstable scenario, the motion-induced loading is further reinforced by the body motion, possibly leading to catastrophic failure. Such unstable interactions involve extraction of energy from the fluid flow such that aerodynamic effects cancel structural damping. Flutter is the term given to this unstable situation, which is a common design issue for long-span bridges.

Depending on the phase of the force with respect to the motion, self-excited forces can be associated with the displacement, the velocity, or the acceleration of the structure. Because of these associations, these forces can be thought of as 'aerodynamic contributions' to stiffness, damping, and mass, respectively. In addition to stiffness and damping, aeroelastic effects can couple modes that are not coupled structurally. Whenever the combined aeroelastic action on various modes results in negative damping for a given mode, flutter occurs. By means of structural dynamics considerations and aerodynamic tailoring, flutter must be avoided for the wind velocity range of interest. Even without resulting in flutter, aeroelastic effects can have a significant effect on response.

Wind Tunnel Testing

Despite the obvious advances of computational capabilities over the years, the complexity of the bluff body fluid-structure interaction problems concerning civil engineering structures has precluded numerical solutions for the flow around structures. Thus, wind tunnels remain, at this juncture, the most effective means of estimating wind effects on structures. However, it should be noted that not all structures require wind tunnel testing. For many conventional structures, for example, low-rise buildings, code-based estimates may well suffice. Wind tunnel testing may be necessary, however, when dealing with a novel design or a design for which dynamic and aeroelastic effects are difficult to anticipate. Examples of such structures include, but are not limited to, long-span bridges and tall buildings.

Wind tunnel testing of a given structure first involves appropriate modeling of the wind environment, necessitating various scaling considerations. Geometric scaling is based on the boundary layer height, the scale of turbulence, and the scale of the surface roughness all constrained by the size of the wind tunnel itself. Ideally, these lengths should hold to the same scaling ratio – a performance that can be approached when the boundary layer is simulated over a long fetch with scaled floor roughness. Dynamic scaling requires Reynolds number equality between the wind tunnel and the prototype. Without extraordinary measures, this is most often not possible and must be kept in mind when interpreting results. Velocity scaling is most often obtained from elastic forces of the structure and inertial forces of the flow. Kinematic scaling involves appropriate distributions of the mean velocity and turbulence intensity and can be achieved with flow manipulation in the wind tunnel.

Both active and passive means are available to generate turbulent boundary layers. While active devices such as air jets, flapping vanes and airfoils are capable of generating a wide range of turbulence parameters, passive devices are cheaper and more efficient to implement. Passive devices include spires, fences, grids, and floor roughness. Depending on the length and cross-sectional size of the tunnel, surrounding terrain may also be modeled.

Once an appropriate incident flow has been generated, there are several options for obtaining aerodynamic load data for the structure of interest. Pressure measurements can be performed on the surface of a model, forces can be quantified from the base of a lightweight, rigid model, or forces can be obtained from an aeroelastic model of the structure. Pressure measurements are capable of quantifying localized loading on a structure's surface. Issues such as fatigue loads for cladding panels and panel anchor and glass failure require such localized analysis.

Integrated loads on a structure are often estimated with high-frequency base balances. These devices are generally integrated into a rotating section of the floor of a wind tunnel. A lightweight model of the structure is mounted on the balance for measuring wind loads over a range of incidence angles. The low mass of the model is necessary to ensure that the natural frequency of the model-balance system is well above any expected wind forcing frequency. A primary advantage of this approach is that modal force spectra are obtained directly and can be used in subsequent analytical estimations of building response. As long as the structural geometry does not change, the forces can be used to analyze the effects of internal structural design changes without the need for further wind tunnel tests.

Aeroelastic models allow interaction between structural motion and aerodynamic forces. Such models can be constructed as continuous or discrete models. Continuous models require specialized materials having structural properties matching those of the prototype. Discrete models are simpler to implement and consist of an internal spine to account for structural dynamic feature, with an external cladding that maintains proper geometric scaling with the prototype. Dynamic response of both buildings and bridges can be estimated utilizing such models.

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Surface Activity in Drug Action

In Studies in Interface Science, 2005

2.7.4 Prostaglandins

The prostaglandins are among the most prevalent autacoids and have been detected in almost every tissue and body fluid; they produce, in minute amounts, a remarkably broad spectrum of effects that embrace practically every biological function. No other autacoids show more numerous and diverse effects than do prostaglandins. The CMC values for prostaglandin E1 (3.85 × 10−8 M) and prostaglandin F2α (1.93 × 10−8 M) are reported in literature [250]. Transport through liquid membranes generated by lecithin, cholesterol and lecithin-cholesterol mixtures has been studied in the presence of prostaglandins. The data indicate that prostaglandins in association with cholesterol may be responsible for the aqueous pores present in the lipid bilayers controlling passive transport through biomembranes. The data further indicate that the presence of cholesterol in each of the two constituent monolayers of the lipid bilayer is essential for pore formation by prostaglandins [251].

Colacicco and Basu have reported the correlations between molecular structure and surface function of six prostaglandins in a model membrane system. Using spread films at the air/water interface, they determined surface pressure and surface potential of PGs A1, A2, E1, E2, F1α and F2α All the prostaglandins formed films with low pressure (0 to 9 dynes/cm) and relatively low surface potentials (10 to 250 mV) [252]. Roseman and Yalkowsky have reported the physicochemical properties of prostaglandin F2α (trimethamine salt): solubility behavior, surface properties, and ionization constants [253].

Monomolecular film compression-relaxation behavior was examined for select dinoprost C-15 alkyl esters. Higher homologs of the series such as palmitate and decanoate esters yielded stable expanded monolayers that exhibited minimal relaxation of surface pressure during noncompression. Their limiting molecular areas were consistent with a Hirschfelder model projection in which the prostaglandin moiety assumes a horizontal orientation at the interface with its alkyl ester chain oriented vertical to the surface plane. Shorter chain homologs such as hexanoate, valerate, butyrate, propionate, and acetate also formed expanded monolayers but exhibited increased instability with decreased alkyl chain length, as reflected in their lower surface pressure development during compression and significant relaxation of pressure during noncompression. Such instability can be tied to their increased solubility in the sub-phase solution and higher desorption rate from the interface [254].

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Polymers for Medical Applications

D.W. Hutmacher , in Encyclopedia of Materials: Science and Technology, 2001

5 In Vivo Degradation, Resorption, and Metabolization

In vivo , implant materials are exposed to various body tissues and fluids. The main constituents of body fluids are aqueous solutions containing proteins, enzymes, and salts. In vivo degradation and resorption of aliphatic polyesters is described as a loss of physical and/or chemical integrity resulting from the interaction of a material with living tissue, due to the hydrolysis process. From physical and physiochemical points of view, enzymes which are large molecules cannot penetrate into solid synthetic polymers. Poly(α-hydroxy acids) have ester linkages which are likely to be hydrolyzed by aqueous medium alone or by esterase enzymes. From studies of the effect of an aqueous medium on several suture materials it can be concluded that the degradation and resorption kinetics depend upon the total time spent in an aqueous environment, regardless of whether it is in vivo or in vitro. This implies that the biodegradation process of a bioresorbable polyesters is purely a hydrolytic process and that enzymes have no effect on it. However, these studies show that the degradation by-products during the resorption process may be metabolized by enzymes. It can be concluded that there is no significant enzyme involvement in the early stages of aliphatic polyester degradation and resorption. The involvement can be more pronounced in the later stages however, as erosion and physical fragmentation occur. In contrast, for polymers in the rubbery state, enzymes can play a significant role in their degradation and resorption process.

In general, when in vitro and in vivo degradation and resorption kinetics are compared, or when enzyme-free and enzyme-containing buffer media are compared, as is generally done in the literature, differences can be expected for many reasons other than enzymic degradation of macromolecules. It is often quite difficult to compare exact degradation and resorption kinetics from various independent in vitro and in vivo studies. The rates of in vitro and in vivo degradation and resorption of polymers can be influenced by the factors listed in Table 1. Therefore, bioresorbability and biocompatibility depend very much on the same factors (see also Biodegradation of Polymers ).

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Preliminary Examination of Evidence

Eric Stauffer , ... Reta Newman , in Fire Debris Analysis, 2008

10.4.4 Body Fluids

DNA, like fingerprints, can provide a strong associative link between an item of evidence and an individual. In some samples, body fluids or even skin cells containing DNA may be present. They could become very pertinent to the outcome of the investigation. Exploitation of DNA evidence often starts with a detection and identification of the stains, however, trace levels of DNA, also referred to as contact DNA will not appear as stain. This is then followed by an extraction of biological materials, and finally their analysis. The detection of body fluids usually is performed first by a naked eye observation of the item for obvious stains or discoloration. Then, an alternate light source in the UV range or other selected wavelengths is used. This is followed by a chemical detection of the different body fluids, which consists of applying different chemical reagents on the items and observing either a change of coloration or a chemiluminescence. Once stains have been located, the forensic scientist typically swabs the surface with a small piece of humidified cotton or cuts a piece of the item, which will then be used for confirmatory testing and the subsequent DNA analysis.

The three great enemies of DNA are heat, light, and humidity [12]. All three of these elements are present in a fire. However, even under such conditions, DNA samples have survived fires. Sweet and Sweet report a case in which DNA was extracted from teeth of the incinerated remains of a homicide victim [13]. It also has been shown that even after a fire, blood can be detected using luminol [14]. If the fire is not too intense on the stains, the blood still reacts to the luminol. The same authors noticed that soot and bloodstains looked very alike after a fire and could be confused. They recommended the use of an [14]: "alternate light source (ALS) at 415 nm in conjunction with a Tiffen Orange 21 filter and 580 nm in conjunction with a Tiffen 1 (25) Red filter […]."

When preserving fire debris in airtight containers as recommended for fire debris analysis, the humidity factor may accelerate the degradation of DNA. When heating the sample at 80°C overnight for a passive headspace concentration extraction, DNA will be further degraded. If enough material was present in the sample prior to the manipulation by the fire debris analyst, the DNA extraction and analysis will not necessarily be jeopardized. But it is usually not possible to estimate the amount of nondegraded DNA by just looking at the sample. Therefore, when body fluids may be present in the debris, it is recommended to first observe it with the naked eye under natural and selective light prior to the extraction of ILR. One exception would be if the debris contains highly volatile compounds for which exposure to the air could destroy the ILR content. Fortunately, this kind of situation is not often encountered. And if the ILR are extremely volatile, the extraction can always be carried out at room temperature, thus avoiding the need for heating. The criminalist performing the observation of the item for body fluid stains should do it in the shortest amount of time possible. A good practice would consist of freezing the sample prior to its manipulation, which would decrease the loss of volatile compounds during its examination by lowering the vapor pressure of the liquids. Then, if stains are located, the swabbing or cutting is performed prior to ILR extraction. However, the chemical detection of body fluids on the debris itself is not recommended prior to ILR extraction.

Continuing advances in the science of DNA analysis have resulted in techniques that are extremely sensitive. Consequently, if a DNA analysis is to be conducted on evidence collected from a fire scene, all aspects of its handling and collection should be done with this in mind in order to avoid contamination. Incendiary devices or delay mechanisms are often suitable for DNA analysis, and may provide a useful link between a device used to set the fire and the individual who committed the crime. A properly trained criminalist should be aware of the potential sources of probative DNA from evidence associated with a fire. In any case, when dealing with samples susceptible to containing DNA evidence, the fire debris analyst should contact the appropriate specialist and discuss the situation to prioritize the evidence examination. In the meantime, the debris should be frozen to slow down, if not prevent, the degradation of the potential DNA.

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Medical Applications of Laser Spectroscopy

S. Andersson-Engels , ... S. Svanberg , in Laser Spectroscopy, 1989

1. Medical Analytical Laser Spectroscopy

Several powerful laser spectroscopic techniques now complement conventional atomic absorption and emission spectroscopy techniques which are routinely used for the analysis of constituents in body fluids. Elements of interest may be alkali ions or heavy metals (toxicology). Laser-enhanced ionization (LEI) spectroscopy, resonance ionization spectroscopy (RIS) and resonance ionization mass spectroscopy (RIMS) provide unprecedented sensitivity and accuracy in atomic analysis. Liquid chromatography and capillary zone electrophoresis with fluorescence detection have, in the same way, increased the sensitivity and selectivity in molecular analysis. Fluorescence techniques are also widely used in biology and medicine for immunoassay and DNA sequencing using fluorescent tags. Similar techniques are used in cytofluorometry and automatic cell sorting.

Another technique, which provides real time diagnostics is laser-induced breakdown spectroscopy (LIBS). A beam from a powerful pulsed laser focussed onto tissue gives rise to a hot plasma emitting atomic and ionic lines. Such techniques have been used to characterize the vessel wall in connection with atherosclerotic plaque ablation and for the characterization of gall- and kidney stones in connection with stone lithotripsy.

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