However, for the evaluation of certain proteins or other cellular components, routine formaldehyde fixation and paraffin-based histology are not suitable. Hard tissues are first exposed to different concentrations of ethanol, methyl methacrylate, and benzoyl peroxide for fixation and infiltration, and then are embedded into polyethylene methyl methacrylate (PMMA) along with a polymerization accelerator [37]. The products of VAT are driven to the liver via portal vein system, affecting its function. Raman spectroscopy: Basic principles and applications • Basic principles - Resonance Raman scattering - Surface Enhanced Raman Scattering (SERS) • Instrumentation -Spectrometer - Excitation sources • Raman in catalysis - In situ cells - In situ Raman (of working catalysts) C.V. Raman (1928) micromachining and CD recording are examples of applications of these processes. Gazi et al. Most X-rays have a wavelength ranging from 0.01 to 10 nanometers, corresponding to frequencies in the range 30 petahertz to 30 exahertz (3×1016 Hz to 3×1019 Hz) and energies in the range 100 eV to 100 keV, produced by the deceleration of high-energy … By Khayelihle Ncama, Lembe S. Magwaza, Asanda Mditshwa and Samson Z. Tesfay, By Héctor Herrera Hernández, Adriana M. Ruiz Reynoso, Juan C. Trinidad González, Carlos O. González Morán, José G. Miranda Hernández, Araceli Mandujano Ruiz, Jorge Morales Hernández and Ricardo Orozco Cruz. Publishing on IntechOpen allows authors to earn citations and find new collaborators, meaning more people see your work not only from your own field of study, but from other related fields too. Thermo. Alkohol und Nikotin: Frühintervention, Akutbehandlung und politische Maßnahmen PDF Kindle. The shift in the positions of the bands gives structural information. Control group (black), obese group (BFMI 852–856) (red), obesity related insulin resistant group (BFMI 860–861) (blue). The C-H stretching, amide I-IV, and PO42− anti-symmetric stretching regions were investigated. In future, these novel approaches will shed light on the internal diagnosis of obesity in clinical application. Laser induced fluorescence spectroscopy of human tissues for cancer diagnosis: Cancer is one of the most dreaded diseases of our time and has a very high incidence. Application of u.v. Therefore, disease-induced changes in the position, bandwidth, signal intensity, and area of the spectral bands of the system of interest, can be monitored by IR spectroscopy. Provenance of Mississippian Chert Sword-form Bifaces Illustrating Selection Diversity in Inter- regional Resource Procurement, QUANTIFYING REGIONAL VARIATION IN TERMINAL PLEISTOCENE ASSEMBLAGES FROM THE LOWER TENNESSEE RIVER VALLEY USING CHERT SOURCING, The Use of Dover-Like Tool Stone by Pre-Mississippian Peoples in the Black Bottom of Southern Illinois, “Non-destructive chert sourcing using Fourier Transform Infrared spectroscopy”, A CHERT SOURCING STUDY USING VISIBLE\ NEAR-INFRARED REFLECTANCE SPECTROSCOPY AT THE DOVER QUARRY SITES, TENNESSEE. Obesity might alter the electric activity in brain areas triggering appetite and craving. Disease states are able to induce changes in molecular composition, concentration, structure, and function of biomolecules, which are directly reflected in the vibrational spectral bands, and therefore, they can be evaluated by using vibrational spectroscopy techniques [2]. This instrument employs an interferometer and exploits the well-established mathematical process of Fourier-transformation. Hierarchical clustering of control, obese and obesity related insulin resistant groups in SCAT (A) and VAT (B) in the 1770–1720 cm−1 spectral region. Excess amount of energy is stored in the form of triglycerides in the lipid droplets of adipocytes and result in the increase in the number of adipocyte (hyperplasia) or enlargement in the size of adipocytes (hypertrophy). Clinical Application of NIRS. They differ in their degree of vascularization and innervation. Control group (black), obese group (BFMI 852–856) (red), obesity related insulin resistant group (BFMI 860–861) (blue). General band assignment of the FTIR spectrum of an adipose tissue. PCA analysis successfully operated to groups. The most dramatic lipid-related changes were found for γas(CH2) at 2930 cm−1, whereas the protein secondary changes occurred at the amide I band, for α–helix and β-sheet located at 1660 and 1630 cm−1, respectively. After the embedding procedure, 2-micron tissue sections are obtained with a cryotome [38, 39] and tissue samples are thaw-mounted on IR-transparent CaF2 or BaF2 windows for IR mapping and data processing. When the amount of BAT was decreased, lower unsaturation/saturation ratio, qualitatively longer hydrocarbon acyl chain length of lipids and higher amount of triglycerides were obtained in both adipose tissues of mice lines as seen in Figure 4. Nonalcoholic fatty liver disease (NAFLD) is a frequent lesion associated with obesity, diabetes, and the metabolic syndrome. Since, this analytical technique provides qualitative and quantitative information as a rapid, accurate, noninvasive, cost-effective, and operator-independent for identification of the spectral differences arising from pathological or environmental conditions. The current study clearly revealed the power of FTIR microspectroscopy in the precise determination of obesity-induced structural and functional changes in inguinal and gonadal adipose tissue of mice lines [34]. However, application of IR spectroscopy to obesity research is quite limited so far. These features were more pronounced in a high grade of steatosis. Albanien. Excessive fat accumulation results in changes in cellular and structural remodeling processes and causes alterations in the endocrine and metabolic systems. SCAT possesses higher concentrations of leptin and adiponectin. Pages 21-35 . The ratio of the sample to alkali halide is important; generally, sample to halide ratio is chosen as 1:100. IR spectrometers have been commercially available since the 1940s [16]. Recent innovations in spectroscopy and microspectroscopy contributed to evaluation, characterization, and differentiation of the obesity. In these situations, sections should be embedded in optimum cutting temperature (OCT) medium and cryo-sectioned to retain the most sensitive and undisrupted insight into tissue architecture. PiSystems … It is also employed in forensic analysis in civil and criminal analysis. Before producing an IR sample in solution, a suitable solvent must be chosen. The band positions and bandwidths of the spectral bands are measured from the center of weight (0.80× peak height position). This can be performed by stepwise hydrolysis accompanied with chromatography. In recent years, scientific researchers have continued to increase the performance of this technique on clinical cases such as cancers and metabolic disorders. The disk thickness is important; thin disks are fragile and difficult to handle, while thick disks transmit too little radiation. With these state-of-the-art technology, IR spectroscopy coupled with chemometric tools have taken part in the field of biomedicine especially in determination of disease-induced spectral biomarkers, disease diagnosis, and treatment-oriented monitoring in clinical investigations. Peptides are converted into amino alcohols which are volatile in nature. Available from: Infrared spectroscopy and microscopy in obesity research, Experimental sampling protocol of IR spectroscopy and microscopy, Data analysis of IR spectroscopy and microscopy, Application of IR spectroscopy and microscopy in obesity research, N–H and O–H symmetric stretching: mainly N–H stretching (amide A) of proteins with the little contribution from O–H stretching of polysaccharides, carbohydrates and water, Olefinic = CH stretching vibration: unsaturated lipids, cholesterol esters, Department of Physiology, Faculty of Medicine, Altinbas University, Turkey, Graduate Program in Biomedical Sciences, Institute of Health Sciences, Altinbas University, Turkey, Department of Biophysics, Faculty of Medicine, Altinbas University, Turkey. Various techniques are available for sampling of liquids. Therefore, WAT paves the way for different metabolic diseases and presents great detrimental effect to human health [49, 50]. If there is a distortion in the spectral bands that may be encountered in cell studies, Mie scattering correction should be performed. In another work [74], the potential of FTIR microspectroscopy for grading steatosis on frozen tissue sections was evaluated. The degree of unsaturation was obtained from the ratio of the integrated area of unsaturated lipids (2992–3020 cm−1) to the integrated area of saturated lipids (2830–2980 cm−1), showed stepwise decreases going from colder-exposed to warmer-exposed BAT. In the other study of the same group, lipid profiles in terms of the content and structure of skeletal muscle and adipose tissues were determined to understand better the characteristics of juvenile-onset spontaneous obesity without high fat diet induction. Our readership spans scientists, professors, researchers, librarians, and students, as well as business professionals. The prevalence of obesity has increased drastically in recent decades and reached global epidemic dimensions [3, 4]. The authors are only responsible for the content and writing of this chapter. Finally, the future direction of the field will be mentioned. When a beam of radiation enters a crystal, if the angle of incidence at the interface between the sample and crystal is greater than the critical angle, incident radiation will undergo total internal reflection [33]. In KBr pellet technique, since the system is not exactly in its natural aqueous environment, some alterations in macromolecular structure may occur. The presence of bound water molecules is not only required for the structural stability of the lipid bilayer [27], but also to control the dynamics and structure of lipid bilayer phases [28] and proteins [29, 30]. Other techniques include skinfold thickness, dual energy X-ray absorptiometry, bioelectrical impedance, plethysmography, underwater weighing, computed tomography, and near-IR interactance. For example, the shift in the position of CH2 symmetric and anti-symmetric bands monitors lipid order/disorder, which gives information about the flexibility of the lipid acyl chains [20, 24]. In another study, FTIR spectroscopy was used to determine physio-chemical properties of pectin extracted from the apple pomace that was proposed to inhibit pancreatic lipase (steapsin). Alkali halide disk method is one of the most favorable methods to examine solid samples. Vibrational spectroscopy includes Raman spectroscopy, IR spectroscopy, and Tera Hertz(THz) spectroscopy. There are several issues in obesity area that need to be clarified. As PhD students, we found it difficult to access the research we needed, so we decided to create a new Open Access publisher that levels the playing field for scientists across the world. [72] investigated the biochemical changes caused by cold exposure using FTIR imaging technique in the brown and subcutaneous white adipose tissues (BAT and s-WAT) of 6-week-old C57BL6 mice exposed to different temperatures for 10 days. A variable aperture is placed in this image plane. Especially in ATR mode, samples are directly put on the crystal without any sample preparation step. An important consideration in the choice of IR cells is the type of window material. Seminar on Applications of U.V. Indeed, as the grade of steatosis was lowered, a decrease in glycogen content with a concomitant increase in lipids was observed in comparison to normal liver. The detailed analyses are performed from the subtracted native spectra. Such materials include Diamond, zinc selenide (ZnSe), germanium (Ge), silicon (Si), and thallium/iodide. The rats were fed with diets containing menhaden oil (MO) or coconut oil (CO) for 4 weeks. The shift in the position of the C=O stretching and the PO2− anti-symmetric double stretching bands gives information about hydration status of these functional groups [20, 24]. The risk of diseases is determined according to cutoff values of 102 cm (40 in) for men and 88 cm (35 in) for females (WHO). Cross-linking between RS and COS was confirmed by FTIR spectroscopy analysis. In recent years, since BAT have been proposed as a potential therapeutic target for obesity and related metabolic diseases, targeting BAT, thermogenesis and monitoring BAT metabolism might hold a possible therapeutic potential for these metabolic diseases. Download PDF Copy; View Supplier Profile; Request Quote; Sponsored by Avantes BV Jul 2 2020. Help us write another book on this subject and reach those readers. The dramatic biochemical changes occurred in the nonsteatotic part of the tissue, suggested that the whole tissue reflects the grade of steatosis [74]. The study showed the extracted pectin has potential use in the anti-obesity formulations [79]. Consequently, adipose tissue serves as a main site for activation of the inflammatory response in obesity. In others, you let the sample first absorb light and then measure the wavelength of light emitted. The content of total fat, unsaturated fat, lipid to protein ratio, and collagen and collagen integrity were measured in tissue of interest and the results of IR analysis were revealed differences in the biomolecular composition of adipose and liver tissues among high fat diet-fed BXD RI strains that reflect genetic variation. Adipose tissue depots are classified into white adipose tissue (WAT) and brown adipose tissue (BAT), each of them has unique structure and function. (2020, November 20). Both surface plasmon resonance (SPR) spectroscopy and its counterpart, localized surface plasmon resonance (LSPR) spectroscopy, have been accepted as important means for carrying out not only nanostructure characterization but also label-free chemical and biological sensing. It also allows the detection of lower concentrations of biological components ranging from biomolecules, membranes, cells, tissues, and body fluids. Infrared spectroscopy is widely used in industry as well as in research. Each PC describes the spectral variability among samples in decreasing order. Chemical shifts (what type is it) c. Integration (how many are there) 2. X-Ray Spectroscopy- Principle, Instrumentation and Applications X-rays make up X-radiation, a form of electromagnetic radiation. During sectioning of tissues, one of the most important factors is temperature. Built by scientists, for scientists. Bizeau et al. The IR detector may detect IR light at a single point, a linear array or a focal plane array to view different sections of the sample. PHAR 3152 Pharmaceutical Analysis II APPLICATIONS OF UV/VISIBLE SPECTROSCOPY TO PHARMACEUTICAL QUANTITATIVE ANALYSIS Reference: Pharmaceutical Analysis, A textbook for Pharmacy Students and Pharmaceutical Chemists 2nd Edition, David G. Watson, 2005 Assay Example 1: Furosemide (frusemide) in tablet form A typical example of a straightforward tablet … Based on this fact, vibrational spectroscopy has been considered as a golden tool in the characterization of molecular structure. In vector normalization, the sum of all intensity values squared of normalized mean-centered spectrum is equal to 1. [70] determined the total content of trans fatty acids (TFA) in subcutaneous, retroperitoneal, and visceral fat of morbidly obese and nonobese patients subjected to bariatric surgery or plastic and abdominal surgery. In some cases, light of different wavelengths is shined on a sample and the wavelengths that get absorbed most strongly are measured. BAT is rich in protein content and mitochondria than that of WAT. This study revealed that, FTIR imaging is a promising technique to quantify cold-induced biochemical changes in BAT and s-WAT in a label-free manner [72]. Moreover, useful accessories, such as temperature controller systems and the attenuated total reflectance (ATR) mode are available to increase the fields of applications of FTIR spectroscopy and microscopy. The data suggest that mid-IR spectroscopy combined with statistical analysis allows identifying serum mid-IR signatures that reflect the liver status during NAFLD [73]. Hydrogen Nuclear Magnetic Resonance a. Splitting or coupling (what’s next to what) b. FTIR spectroscopy and synchrotron X-ray fluorescence (SRXRF) microprobe techniques show that brain areas implicated appetite control, upon experimental treatment by either anodal-type tDCS (atDCS) or cathodal-type tDCS (ctDCS). In this method, the two spectra with the highest similarity (i.e., spectra with the smallest spectral distance) are merged into a cluster and then the distances between this cluster and all other spectra are calculated. APPLICATIONS OF IR SPECTROSCOPY. APPLICATIONS OF RAMAN SPECTROSCOPY by ROSEMARY SANCHES Submitted to the Department of Biology on August 1977 in partial fulfillment of the requirements for the Degree of Master of Science ABSTRACT I The frequency of vibration of the C = 0 group in dimethylformamide, N-methylformamide, N-methylacetamide, acetone and methylacetate has been measured in a number of solvents. Increased circulating levels of inflammatory markers are associated with obesity-related pathologies. The absorbance in the spectral maps was represented in color-coded images, where low absorption was represented in blue and high absorption was represented in red color. In order to remove any artifact that arise from the thickness of the sample, generally in cell and tissue studies, area ratios of the specific bands are used [41, 42]. Our team is growing all the time, so we’re always on the lookout for smart people who want to help us reshape the world of scientific publishing. The infiltration and embedding medium for soft tissues is paraffin. This could originate from it possessing a lower amount of brown adipose tissue. How? Some of the major applications of IR spectroscopy are as follows: 1. IR spectroscopy is a technique based on the study of absorption of IR radiation, which causes transition between vibrational energy states in the molecule. Spectral differences indicated differences in lipid structure and content of BFMI lines, which may originate from different insulin sensitivity levels of the lines. Both genetic and environmental factors contribute to the development of metabolic complications, which result in a significant health and economic burden. This inflammatory state is associated with increased accumulation of macrophages in adipose tissues along with the production of inflammatory cytokines and their secretion into the circulation (e.g., adiponectin, leptin, resistin, IL6, IL10, TNFα, macrophage migration inhibitory factor [51], and components of the renin-angiotensin system [52]). Obesity is one of the main factors that increases the risk of many diseases and contributes to functional disabilities in tissues such as adipose, liver, and muscle. The authors report no financial conflicts of interest. Since the SCAT is more accessible than the VAT for medical interventions, this type of adipose tissue can be used preferably in biopsies and bariatric operations [43]. The most significant advance in IR spectroscopy is the introduction of Fourier-transform methodology into the spectrometers. Fatty liver, itself, stimulates a local subacute inflammatory state by production of inflammatory mediators such as TNFα or IL-6 that contribute to hepatic and systemic insulin resistance [62]. Detection of Impurities UV absorption spectroscopy is one of the best methods for determination of impurities in organic molecules. It is a well-known technique for reducing the dimensionality of multivariate data while preserving most of the variance [43]. Mass spectroscopy has an important application in analysis of sequence of amino acids in proteins and peptides, that is, analysis of structure of proteins and peptides, and this is employed increasingly. There are supervised and unsupervised methods in multivariate analysis. It is mandatory to correct differences in spectral intensity that arises from heterogeneity in sample thickness or inaccuracies in positioning. Infrared (IR) spectroscopy is a vibrational spectroscopic technique based on the absorption of infrared radiation by matters that excite vibrations of molecular bonds. 08028 Barcelona. If absorbed energy is plotted as a function of wavenumber, an IR spectrum is observed. Furthermore, FTIR spectra may give information about the disease-induced changes at very early stage, earlier than they become visible. In addition, with constant improvements in computers, and coupling with microscope to the system, IR spectroscopy has made further great strides. Biospectroscopy is an analytical multistep process, including sample preparation, spectral acquisition, pre-processing, and computational analysis. TFA were measured by FTIR-ATR spectroscopy. The secretions of SCAT are released into the general circulation [48]. Complementary H-1 NMR measurements confirmed the findings from this ratio in BAT. To this end we have integrated the discus-sion of IR and Raman group frequencies into different classes of organic groups. These properties can be used for obtaining functional group information or monitoring molecules in different conditions such as disease states. ATR is accepted to require little or no sample preparation, and is one of the most versatile sampling techniques. Bortolotto et al. The findings provided evidence for the development of IR biochemical obesity markers and showed the potential of FTIR microspectroscopy in the evaluation of the effectiveness of the drug in the treatment of obese patients [33]. UV Spectroscopy- Principle, Instrumentation, Applications. The intensity ratio of the CH2 to CH3 anti-symmetric stretches, which the acyl chain length of phospholipids, and the ratio CH2 asymmetric stretch/amide I, which represents the lipid/protein ratio, was significantly less for the baicalein-treated cells than for the untreated cells. By making research easy to access, and puts the academic needs of the researchers before the business interests of publishers. Different kinds of embedding media can be used for soft and hard tissues. GRAMS/AI Spectroscopy Software... provides a set of complementary, fully integrated applications that increase productivity across all areas of spectroscopic analysis. First, visual inspection of the spectra should be done to remove bad quality spectra. In the case of nutrition overload, function of ER is impaired, protein folding is disturbed, ER stress develops and activates a signaling network called the unfolded protein response (UPR) to restore ER homeostasis [64, 65]. Masatsugu Niwayama, Yutaka Yamashita. Then, mixture is ground with an agate mortar and subjected to a pressure of 10 ton in-2 (1.575 x 105 kg m-2) in an evacuated die. The discriminatory power of these techniques in diagnosis of obesity will be discussed. Aboualizadeh et al. This sinters the mixture and produces clear transparent disks. [78] evaluated nano lipo-vesicles phytosomal thermogel of Soybean [Glycine max (L.) Merrill], for its anti-obesity action on body weight gain, adipose tissue size, and lipid profile data by FTIR spectroscopy. The muscle (longissimus, quadriceps) and adipose (inguinal, gonadal) tissues of 10-week-old male DBA/2 J and Berlin Fat Mouse inbred (BFMI) lines (BFMI856, BFMI860, BFMI861) fed with a standard breeding diet were used. IR spectroscopy products measure the transition between stretching vibrations (symmetrical and anti-symmetrical types) and bending vibrations. It is a powerful method for investigating structural, functional, and compositional changes in biomolecules, cells, and tissues. The waist circumference (WC) is also used to monitor central obesity and measured at a level midway between the lowest rib and the iliac crest. The KBr pellet technique has been extensively used to study a variety of biological systems in dry stated [31, 32]. A new absorption peak at 1560.88 cm−1 appeared in the COS-RS complex spectra, and the shape of typical absorption peaks for starch from 1180 to 953 cm−1 also changed. In the literature, visceral adipose tissue has been implicated as a major risk factor for insulin resistance, type 2 diabetes [48, 58], cardiovascular disease [59], stroke [60], and metabolic syndrome [61]. Brief introduction to this section that descibes Open Access especially from an IntechOpen perspective, Want to get in touch? Near-infrared spectroscopy (NIRS) is a spectroscopic method that uses the near-infrared region of the electromagnetic spectrum (from 780 nm to 2500 nm). Different from other methods, Ward’s algorithm tries to find as homogeneous groups as possible. There are several orders of derivative; first order is according to slope of the peak and second order is according to curvature radius. To date our community has made over 100 million downloads. Due to its technical problems such as the difficulty in determining bony prominences, this method identifies obesity risk poorly. Spectroscopy is based on the interactions between matter (sample analyzed) and energy from the portion of electromagnetic spectrum. The bandwidth of the CH2 stretching bands gives lipid fluidity information. PCA can be used to define groups based on spectral similarity/variance as classification method. The results suggested that the consumption of COS-RS complexes exerted a more efficient recovery of insulin sensitivity compared to individual treatments [77]. The adipose tissue samples were obtained from 10 weeks old male control (DBA/2 J) (n = 6) and four different obese BFMI mice lines (n = 6 per group). The other factors while choosing solvent are that it should be as nonpolar as possible to minimize solute–solvent interactions; and it should not strongly absorb IR radiation. Application Of Nmr Spectroscopy.pdf - Free download Ebook, Handbook, Textbook, User Guide PDF files on the internet quickly and easily. Fundamentals and Applications of Laser Induced Breakdown Spectroscopy. It is applied before data analysis but should be kept minimum not to introduce some artifacts and should be performed in the right order. PDF generated at: Sat, 30 May 2009 17:33:36 UTC Spectroscopy: Principles, Theory, Techniques and Applications. For COS analysis, in IR spectroscopy, the absorption peak at 3416.33 cm−1 belonged to O-H and N-H stretching vibrations, 2922.80 cm−1 corresponding to a C-H stretching vibration, and 1566.70 cm−1 originated from -NH2 absorption peak were used. Usually, the spacer is made of polytetrafluoroethylene (PTFE, Teflon) with a variety of thickness is used. As seen from Figure 3, successful discrimination of the obese, obesity-related insulin resistant and control groups was achieved with high sensitivity and specificity. Rsad2, which may modulate lipid droplet contents and lipid biosynthesis, and Colec11, which may play a role in apoptotic cell clearance and maintenance of adipose tissue were suggested as potential quantitative trait candidate genes. Spectroscopy NMR, IR, MS, UV-Vis Main points of the chapter 1. A drop of liquid is sandwiched, using suitable spacer, between two IR windows that are then mounted in a cell holder. Stability of body fat is key point to maintain metabolic homeostasis. This procedure is repeated until only one big cluster is left. This spectrum is quite complex and contains several bands, which arise from different functional groups belonging to biomolecules of the system. Furthermore, PCA is capable to obtain loadings plots to identify the main contributory variables (i.e., wavenumbers) that determine the discriminating wavenumbers throughout the IR spectrum. When the applied IR energy is equal to the energy difference between vibrational states, absorption of IR radiation takes place and a peak is observed. In this section, the basic ideas and definitions associated with IR spectroscopy and microscopy will be described. It is known that ER stress is linked with obesity-related pathologies including insulin resistance, type 2 diabetes, hypertension, and nonalcoholic fatty liver disease [5, 65, 66]. 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