Development of a method based on ATR-FTIR spectroscopy to detect maple syrup adulterated with added syrups.

BACKGROUND: Food adulteration is a global concern, whether it takes place intentionally or incidentally. In Canada, maple syrup is susceptible to being adulterated with cheaper syrups such as corn, beet, cane syrups, and many more due to its high price and economic importance. RESULTS: In this study, the use of attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy was investigated to detect maple syrups adulterated with 15 different sugar syrups at different concentration levels. The spectra were collected in the range of 4000-650 cm-1 in the absorbance unit. These spectra were used to build six libraries and three models. A method that is capable of performing a qualitative library search using a similarity search, which is based on the first derivative correlation search algorithm, was developed. This method was further evaluated and proved to be able to capture adulterated and reject non-adulterated maple syrups, belonging to the color grades golden and amber maple syrups, with an accuracy of 93.9% and 92.3%, respectively. However, for the maple syrup belonging to the dark color grade, this method demonstrated low specificity of 33.3%, and for this reason it was only able to adequately detect adulterated samples from the non-adulterated ones with an accuracy of 81.4%. CONCLUSION: This simple and rapid method has strong potential for implementation in different stages of the maple syrup supply chain for early adulteration detection, particularly for golden and amber samples. Further evaluation and improvements are required for the dark color grade. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Multicenter Evaluation of Attenuated Total Reflectance Fourier Transform Infrared (ATR-FTIR) Spectroscopy-Based Method for Rapid Identification of Clinically Relevant Yeasts.

Fourier transform infrared (FTIR) spectroscopy has demonstrated applicability as a reagent-free whole-organism fingerprinting technique for both microbial identification and strain typing. For routine application of this technique in microbiology laboratories, acquisition of FTIR spectra in the attenuated total reflectance (ATR) mode simplifies the FTIR spectroscopy workflow, providing results within minutes after initial culture without prior sample preparation. In our previous central work, 99.7% correct species identification of clinically relevant yeasts was achieved by employing an ATR-FTIR-based method and spectral database developed by our group. In this study, ATR-FTIR spectrometers were placed in 6 clinical microbiology laboratories over a 16-month period and were used to collect spectra of routine yeast isolates for on-site identification to the species level. The identification results were compared to those obtained from conventional biochemical tests and/or matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Isolates producing discordant results were reanalyzed by routine identification methods, ATR-FTIR spectroscopy, and PCR gene sequencing of the D1/D2 and internal transcribed spacer (ITS) regions. Among the 573 routine clinical yeast isolates collected and identified by the ATR-FTIR-based method, 564 isolates (98.4%) were correctly identified at the species level, while the remaining isolates were inconclusive with no misidentifications. Due to the low prevalence of Candida auris in routine isolates, additional randomly selected C. auris (n = 24) isolates were obtained for evaluation and resulted in 100% correct identification. Overall, the data obtained in our multicenter evaluation study using multiple spectrometers and system operators indicate that ATR-FTIR spectroscopy is a reliable, cost-effective yeast identification technique that provides accurate and timely (∼3 min/sample) species identification promptly after the initial culture.

Evaluation of Two Portable Hyperspectral-Sensor-Based Instruments to Predict Key Soil Properties in Canadian Soils.

In contrast with classic bench-top hyperspectral (multispectral)-sensor-based instruments (spectrophotometers), the portable ones are rugged, relatively inexpensive, and simple to use; therefore, they are suitable for field implementation to more closely examine various soil properties on the spot. The purpose of this study was to evaluate two portable spectrophotometers to predict key soil properties such as texture and soil organic carbon (SOC) in 282 soil samples collected from proportional fields in four Canadian provinces. Of the two instruments, one was the first of its kind (prototype) and was a mid-infrared (mid-IR) spectrophotometer operating between ~5500 and ~11,000 nm. The other instrument was a readily available dual-type spectrophotometer having a spectral range in both visible (vis) and near-infrared (NIR) regions with wavelengths ranging between ~400 and ~2220 nm. A large number of soil samples (n = 282) were used to represent a wide variety of soil textures, from clay loam to sandy soils, with a considerable range of SOC. These samples were subjected to routine laboratory soil analysis before both spectrophotometers were used to collect diffuse reflectance spectroscopy (DRS) measurements. After data collection, the mid-IR and vis-NIR spectra were randomly divided into calibration (70%) and validation (30%) sets. Partial least squares regression (PLSR) was used with leave one out cross-validation techniques to derive the spectral calibrations to predict SOC, sand, and clay content. The performances of the calibration models were reevaluated on the validation set. It was found that sand content can be predicted more accurately using the portable mid-IR spectrophotometer and clay content is better predicted using the readily available dual-type vis-NIR spectrophotometer. The coefficients of determination (R2) and root mean squared error (RMSE) were determined to be most favorable for clay (0.82 and 78 g kg-1) and sand (0.82 and 103 g kg-1), respectively. The ability to predict SOC content precisely was not particularly good for the dataset of soils used in this study with an R2 and RMSE of 0.54 and 4.1 g kg-1. The tested method demonstrated that both portable mid-IR and vis-NIR spectrophotometers were comparable in predicting soil texture on a large soil dataset collected from agricultural fields in four Canadian provinces.

Diagnosis of idiopathic amyotrophic lateral sclerosis using Fourier-transform infrared spectroscopic analysis of patient-derived skin.

One of the great challenges in identifying effective therapy in many neurodegenerative diseases, such as amyotrophic lateral sclerosis (ALS), is the lack of reliable biomarkers. In this study, we applied infrared imaging microspectroscopy, a valuable technique to investigate biomolecule fingerprints and secondary structure of proteins within biological tissue. We hypothesized that, since skin and CNS have the same embryonic origin, spectral differences associated with ALS-specific pathological events will be readily detectable through skin testing using this technique. Cells from healthy individuals and ALS patients were isolated from skin biopsies in order to generate tissue-engineered in vitro skin (TES). Infrared spectra of the generated TES were recorded using a focal-plane-array Fourier transform infrared (FPA-FTIR) spectrometer, and hierarchical cluster analysis of the spectral data was performed in order to establish clear differences between the tested TES specimens. Interestingly, our analyses showed that it was readily possible to discriminate ALS- and control-TES solely based on differences in associated FTIR spectra, mainly located between 1149 and 1473 cm-1, attributed to disruption of phospholipid cell membranes, extracellular matrix remodeling or cholesterol accumulation. Spectral differences within the TES samples may therefore be associated with disease state, paving the way for the identification of biomarkers in ALS.

Reagent-Free Identification of Clinical Yeasts by Use of Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy.

Invasive fungal infections by opportunistic yeasts have increased concomitantly with the growth of an immunocompromised patient population. Misidentification of yeasts can lead to inappropriate antifungal treatment and complications. Attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy is a promising method for rapid and accurate identification of microorganisms. ATR-FTIR spectroscopy is a standalone, inexpensive, reagent-free technique that provides results within minutes after initial culture. In this study, a comprehensive spectral reference database of 65 clinically relevant yeast species was constructed and tested prospectively on spectra recorded (from colonies taken from culture plates) for 318 routine yeasts isolated from various body fluids and specimens received from 38 microbiology laboratories over a 4-month period in our clinical laboratory. ATR-FTIR spectroscopy attained comparable identification performance with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). In a preliminary validation of the ATR-FTIR method, correct identification rates of 100% and 95.6% at the genus and species levels, respectively, were achieved, with 3.5% unidentified and 0.9% misidentified. By expanding the number of spectra in the spectral reference database for species for which isolates could not be identified or had been misidentified, we were able to improve identification at the species level to 99.7%. Thus, ATR-FTIR spectroscopy provides a new standalone method that can rival MALDI-TOF MS for the accurate identification of a broad range of medically important yeasts. The simplicity of the ATR-FTIR spectroscopy workflow favors its use in clinical laboratories for timely and low-cost identification of life-threatening yeast strains for appropriate treatment.

Development of an NDIR CO2 sensor-based system for assessing soil toxicity using substrate-induced respiration.

The eco-toxicological indicators used to evaluate soil quality complement the physico-chemical criteria employed in contaminated site remediation, but their cost, time, sophisticated analytical methods and in-situ inapplicability pose a major challenge to rapidly detect and map the extent of soil contamination. This paper describes a sensor-based approach for measuring potential (substrate-induced) microbial respiration in diesel-contaminated and non-contaminated soil and hence, indirectly evaluates their microbial activity. A simple CO2 sensing system was developed using an inexpensive non-dispersive infrared (NDIR) CO2 sensor and was successfully deployed to differentiate the control and diesel-contaminated soils in terms of CO2 emission after glucose addition. Also, the sensor system distinguished glucose-induced CO2 emission from sterile and control soil samples (p ≤ 0.0001). Significant effects of diesel contamination (p ≤ 0.0001) and soil type (p ≤ 0.0001) on glucose-induced CO2 emission were also found. The developed sensing system can provide in-situ evaluation of soil microbial activity, an indicator of soil quality. The system can be a promising tool for the initial screening of contaminated environmental sites to create high spatial density maps at a relatively low cost.

Dapagliflozin diminishes memory and cognition impairment in Streptozotocin induced diabetes through its effect on Wnt/ß-Catenin and CREB pathway.

Diabetic neuropathy is a chronic condition that affects a significant number of individuals with diabetes. Streptozotocin injection intraperitoneally to rodents produces pancreatic islet ß-cell destruction causing hyperglycemia, which affect the brain leading to memory and cognition impairment. Dapagliflozin may be able to reverse beta-cell injury and alleviate this impairment. This effect may be via neuroprotective effect or possible involvement of the antioxidant, and anti-apoptotic properties. Forty rats were divided into four groups as follows: The normal control group, STZ-induced diabetes group, STZ-induced diabetic rats followed by treatment with oral dapagliflozin group and normal rats treated with oral dapagliflozin. Behavioral tests (Object location memory task and Morris water maze) were performed. Serum biomarkers (blood glucose and insulin) were measured and then the homeostatic model assessment for insulin resistance (HOMA-IR) was calculated. In the hippocampus the followings were determined; calmodulin, ca-calmodulin kinase Ⅳ (CaMKIV), protein kinase A (PKA) and cAMP-responsive element-binding protein to determine the transcription factor CREB and its signaling pathway also Wnt signaling pathway and related parameters (WnT, B-catenin, lymphoid enhancer binding factor LEF, glycogen synthase kinase 3ß). Moreover, nuclear receptor-related protein-1, acetylcholine and its hydrolyzing enzyme acetylcholine esterase, oxidative stress parameter malondialdehyde (MDA) and apoptotic parameter caspase-3 were determined. STZ was able to cause destruction to pancreatic ß-cells which was reflected on glucose levels causing diabetes. Diabetic neuropathy was clear in the rats performing the behavioral tests. Memory and cognition parameters in the hippocampus were negatively affected. Oxidative stress and apoptotic parameter were elevated while the electrical activity was declined. Dapagliflozin was able to reverse the previously mentioned parameters and behavior. Thus, to say dapagliflozin significantly showed neuroprotective action along with antioxidant, and anti-apoptotic properties.

Fourier Transform Infrared Spectroscopy as a Tool to Study Milk Composition Changes in Dairy Cows Attributed to Housing Modifications to Improve Animal Welfare.

Animal welfare status is assessed today through visual evaluations requiring an on-farm visit. A convenient alternative would be to detect cow welfare status directly in milk samples, already routinely collected for milk recording. The objective of this study was to propose a novel approach to demonstrate that Fourier transform infrared (FTIR) spectroscopy can detect changes in milk composition related to cows subjected to movement restriction at the tie stall with four tie-rail configurations varying in height and position (TR1, TR2, TR3 and TR4). Milk mid-infrared spectra were collected on weekly basis. Long-term average spectra were calculated for each cow using spectra collected in weeks 8-10 of treatment. Principal component analysis was applied to spectral averages and the scores of principal components (PCs) were tested for treatment effect by mixed modelling. PC7 revealed a significant treatment effect (p = 0.01), particularly for TR3 (configuration with restricted movement) vs. TR1 (recommended configuration) (p = 0.03). The loading spectrum of PC7 revealed high loadings at wavenumbers that could be assigned to biomarkers related to negative energy balance, such as ß-hydroxybutyrate, citrate and acetone. This observation suggests that TR3 might have been restrictive for cows to access feed. Milk FTIR spectroscopy showed promising results in detecting welfare status and housing conditions in dairy cows.

Antimicrobial activity of various essential oils and their application in active packaging of frozen vegetable products.

Essential oils (EOs) have potential utility as clean-label food preservatives due to their antimicrobial and antioxidant properties. In this study, various EOs were screened for their antimicrobial activities against Listeria grayi in vitro. The susceptibility of L. monocytogenes to select EOs was compared with that of L. grayi. The effectiveness of the selected EOs in inhibiting the growth of L. grayi on vegetable products was also investigated. The results showed that cinnamon and oregano EOs and carvacrol were effective in the vapor phase in inhibiting the growth of L. grayi as well as L. monocytogenes, with the susceptibility of L. monocytogenes to cinnamon EO being slightly higher than that of L. grayi. The packaging of green peppers with cellulose stickers impregnated with cinnamon EO at 556 µL/Lheadspace reduced the Listeria count to 1 log CFU/g after 2 days of storage as compared to 7.5 log CFU/g for controls.

Effect of the homogenization technique on the enumeration of psychrotrophic bacteria in food absorbent pads.

Four methods were tested for enumerating bacteria present in the absorbent pads (AP) used in packaging chicken and other meats. Viable counts were ascertained at day 0 and day 7 (d0 and d7, respectively). Sampling bacterial cells from AP were carried out using a countertop blender, Stomacher, sonication, and blender in combination to sonication. The release of bacterial cells by breaking down the AP with the blender resulted in the highest CFU counts. At d0, a bacterial recovery rate of 94% was obtained with the blender, while the recovery rates using Stomacher or sonication alone were 58% and 73%, respectively. At d7, the Stomacher treatment also gave the lowest colony forming unit (CFU) values in the AP incubated at 7 °C. Sonication of the AP prior to homogenization with the blender did not increase CFU counts. Results suggested that breaking down the AP with a blender gives higher CFU levels than the Stomacher, which is the most commonly used technique for this purpose.

Protein-biomolecule interactions play a major role in shaping corona proteome: studies on milk interacted dietary particles.

Despite the significance of surface absorbed proteins in determining the biological identity of nanoparticles (NPs) entering the human body, little is known about the surface corona and factors that shape their formation on dietary particles used as food additives. In this study, food grade NPs of silica and titania and their food additive counterparts (E551 and E171) were interacted with milk proteins or with skimmed milk and the levels of protein adsorption were quantified. Characteristics of proteins correlating with their level of adsorption to NPs were determined using partial least squares regression analysis. Results from individual protein-particle interactions revealed the significance of factors such as zeta potential, hydrophobicity and hydrodynamic size of particles, and protein characteristics such as the number of beta strands, isoelectric points, the number of amino acid units (Ile, Tyr, Ala, Gly, Pro, Asp, and Arg), and phosphorylation sites on their adsorption to particles. Similar regression analysis was performed to identify the characteristics of twenty abundant and enriched proteins (identified using LC-MS/MS analysis) for their association with the surface corona of milk-interacted particles. Contrary to individual protein-particle interactions, protein characteristics such as helices, turns, protein structures, disulfide bonds, the number of amino acid units (Cys, Met, Leu, and Trp), and Fe binding sites were significant for their association with the surface corona of milk interacted particles. This difference in factors identified from individual proteins and milk interacted particles suggested possible interactions of proteins with surface adsorbed biomolecules as revealed by scanning transmission X-ray microscopy and other biochemical assays.

Cellulose Nanocrystals Derived from Textile Waste through Acid Hydrolysis and Oxidation as Reinforcing Agent of Soy Protein Film.

More than 10 million tons of textile waste are disposed through landfill every year in North America. The disposal of textile waste via landfill or incineration causes environmental problems and represents a waste of useful resources. In this work, we explored the possibility to directly extract cellulose nanocrystals (CNCs) from untreated textile waste through two methods, namely sulfuric acid hydrolysis and three-step oxidization. CNCs with cellulose Iß crystalline structure and rod-like shape were successfully obtained. The aspect ratios of CNCs prepared from acid hydrolysis and oxidization were 10.00 ± 3.39 and 17.10 ± 12.85, respectively. Their application as reinforcing agent of soybean protein isolate (SPI) film was evaluated. With the addition of 20% CNCs, the composite film maintained the high transparency, while their water vapor barrier property, tensile strength, and Young's modulus were significantly improved. This research demonstrates a promising approach to recycle textile waste, and more value-added applications based on the derived CNCs could be expected.

The influence of SLC22A1 rs622342 and ABCC8 rs757110 genetic variants on the efficacy of metformin and glimepiride combination therapy in Egyptian patients with type 2 diabetes.

Background: The incidence of Type 2 Diabetes Mellitus (T2DM) in Egypt is considered one of the highest in the world. Metformin and Sulfonylureas are usually prescribed together due to their efficacy and their relatively low cost. Organic cation transport 1, encoded by SLC22A1 gene, is the main transporter of metformin into hepatocytes, which is considered metformin site of action. Sulfonylureas enhance insulin release from pancreatic B-cells through binding to sulfonylurea receptor 1, encoded by ABCC8 gene. Single nucleotide polymorphisms in the SLC22A1 and ABCC8 genes might affect the response of each drug. Aims: To investigate the influence of SLC22A1 rs622342 (A>C) and ABCC8 rs757110 (A>C) genetic variants on the efficacy of metformin and glimepiride combination therapy in Egyptian T2DM patients. Methods: Observational cross-sectional study in which patients receiving metformin and glimepiride combination therapy for at least 6 months were included for genotyping and classified into either responders or non-responders, based on their HbA1C level. Results: A total of 127 patients were included and genotyped. They were divided into 93 responders (HbA1C<7%) and 34 non-responders (HbA1C≥7%). Minor allele frequencies for rs622342 and rs757110 were 0.189 and 0.271, respectively. Only SLC22A1 rs622342 variant was found to be associated with the response of combination therapy, in which AA alleles carriers were 2.7-times more responsive to metformin than C allele carriers (Recessive model, odds ratio = 2.718, p = 0.025, 95% CI = 1.112-6.385). Conclusion: Genotyping of rs622342 can be useful in predicting the response to metformin in combination therapy in Egyptian T2DM patients.

A Comparative Analysis of Different Grades of Silica Particles and Temperature Variants of Food-Grade Silica Nanoparticles for Their Physicochemical Properties and Effect on Trypsin.

While silica particles are used extensively in food products, different grades and temperature variants of silica particles have not been compared for their physiochemical and biological properties. Different grades of silica (food-grade nanoparticles (FG-NPs), nonfood-grade nanoparticles (NFG-NPs), and food-grade micron particles (FG-MPs)) and the temperature variants generated by exposing FG-NPs to wet heating, dry heating, and refrigeration were compared for their physicochemical properties and interaction with trypsin. FG-NPs were similar to NFG-NPs and FG-MPs in their elemental composition and amorphous nature but had relatively less branched and ring siloxane groups than the latter ones. There were subtle but noticeable changes in the agglomeration behavior and relative abundance of different silica groups in FG-NPs exposed to food-handling temperatures. Secondary structure and function of trypsin were negatively impacted by FG-NPs and their temperature variants. Silica particles showed a "mixed-type inhibition" of trypsin resulting in partial digestion of bovine serum albumin. In conclusion, our studies showed differences in the surface chemistry of different grades of silica particles and temperature variants of FG-NPs and their negative impact on the structure and function of trypsin.

Cell Seeding on UV-C-Treated 3D Polymeric Templates Allows for Cost-Effective Production of Small-Caliber Tissue-Engineered Blood Vessels.

There is a strong clinical need to develop small-caliber tissue-engineered blood vessels for arterial bypass surgeries. Such substitutes can be engineered using the self-assembly approach in which cells produce their own extracellular matrix (ECM), creating a robust vessel without exogenous material. However, this approach is currently limited to the production of flat sheets that need to be further rolled into the final desired tubular shape. In this study, human fibroblasts and smooth muscle cells were seeded directly on UV-C-treated cylindrical polyethylene terephthalate glycol-modified (PETG) mandrels of 4.8 mm diameter. UV-C treatment induced surface modification, confirmed by Fourier-transform infrared spectroscopy (FTIR) analysis, was necessary to ensure proper cellular attachment and optimized ECM secretion/assembly. This novel approach generated solid tubular conduits with high level of cohesion between concentric cellular layers and enhanced cell-driven circumferential alignment that can be manipulated after 21 days of culture. This simple and cost-effective mandrel-seeded approach also allowed for endothelialization of the construct and the production of perfusable trilayered tissue-engineered blood vessels with a closed lumen. This study lays the foundation for a broad field of possible applications enabling custom-made reconstructed tissues of specialized shapes using a surface treated 3D structure as a template for tissue engineering.

Evaluation of Fourier transform infrared spectroscopy for the rapid identification of glycopeptide-intermediate Staphylococcus aureus.

OBJECTIVES: To evaluate Fourier transform infrared (FTIR) spectroscopy as a rapid method for distinguishing glycopeptide-intermediate Staphylococcus aureus (GISA) from glycopeptide-susceptible methicillin-resistant S. aureus (MRSA) and to compare three data analysis methods. METHODS: First-derivative normalized spectra of dried films of bacterial growth on Que-Bact Universal Medium No. 2 were examined by singular value decomposition to identify key spectral regions. Region selection was analysed by principal component analysis (PCA), self-organizing maps (SOMs) and the K-nearest neighbour (KNN) algorithm. The initial data set included 35 GISA (including GISA Mu50 and heterogeneous GISA Mu3) and 25 epidemic MRSA. The regions were then tested using enlarged data sets that included 22 sporadic and 85 additional epidemic MRSA. RESULTS: Epidemic MRSA and GISA/hGISA were separated into two distinct clusters on the basis of spectral data from regions 1352-1315 and 1480-1460 cm(-1), the former providing 100% correct classification by all three analyses and the latter providing 96.67% correct by PCA, 98.34% by SOM and 100% by KNN. The 1480-1460 cm(-1) region was more effective for distinguishing GISA/hGISA from a set combining sporadic and epidemic MRSA, with two GISA/hGISA and four sporadic MRSA misclassified by PCA and SOM (92.69% correct), while the KNN method misclassified three of the four sporadic MRSA (93.90% correct). The addition of 85 other epidemic MRSA this set increased the fraction of correctly classified isolates to 96.41% and 97.01% by PCA, SOM and KNN, respectively. CONCLUSIONS: As only 6 of 167 isolates were misclassified, FTIR spectroscopy may provide means of rapid and accurate identification of GISA and hGISA among isolates of MRSA.

Epidemiological typing of methicillin-resistant Staphylococcus aureus strains by Fourier transform infrared spectroscopy.

A rapid and simple typing system is needed for controlling the spread of epidemic methicillin-resistant Staphylococcus aureus (MRSA), currently one of the most widespread multi-resistant nosocomial pathogens in Canadian hospitals. Fourier transform infrared (FTIR) spectroscopy was used to subtype 85 isolates representing five strains of epidemic Canadian MRSA (CMRSA). Spectral fingerprints of whole cells grown on Que-Bact(R) Universal Medium No. 2 were transformed to first derivative peak-height normalized files and examined visually and by singular-value decomposition (SVD). Distinguishing spectral regions were processed by principal component analysis (PCA), self-organizing map and K-nearest neighbor supervised cluster analysis. Among the visually identified regions, 1070-1050 and 1155-1137 cm(-1) were found suitable for discrimination of CMRSA-4 and CMRSA-2 respectively, while CMRSA-1, CMRSA-3, and CMRSA-5 each exhibited distinctive spectral profiles in the 1123-1094 cm(-1) region. The combination, 1123-1094, 1174-1154 and 2904-2864 cm(-1) separated the five CMRSA with 84.6% correct classification by PCA. Five clusters were also obtained using the SVD-selected regions 1096-1066, 1118-1090 and 2914-2880 cm(-1), with 87.8% correct classification based on visual examination of the PCA scores plot and 97% based on supervised cluster analysis. These results demonstrate that FTIR spectroscopy has considerable potential as a rapid (1-hour) and simple method for MRSA strain typing and monitoring in clinical settings.

Rapid identification of coagulase-negative staphylococci by Fourier transform infrared spectroscopy.

Coagulase-negative staphylococci (CNS), frequently associated with both community-acquired and nosocomial bloodstream infections, must be distinguished from Staphylococcus aureus for clinical purposes. Conventional methods are too laborious and time-consuming and often lack sensitivity to CNS. Fourier transform infrared (FTIR) spectroscopy combined with the use of a universal growth medium (Que-Bact Universal Medium No. 2) and chemometrics was evaluated for its potential as a rapid and simple clinical tool for making this distinction. FTIR spectra of 11 methicillin-sensitive and 11 methicillin-resistant CNS isolates as well as 25 methicillin-sensitive, 47 methicillin-resistant, 34 borderline oxacillin-resistant and 35 glycopeptide intermediate S. aureus isolates were obtained from dried films of stationary-phase cells grown on the universal medium. Principal component analysis (PCA), self-organizing maps, and the K-nearest neighbor algorithm were employed to cluster the different phenotypes based on similarity of their FTIR spectra. PCA of the first-derivative normalized spectral data from a single narrow region (2888-2868 cm(-1)) yielded complete differentiation of CNS from both methicillin-sensitive and methicillin-resistant S. aureus. The rate of correct classification was somewhat reduced, from 100% to 90%, after inclusion of borderline oxacillin-resistant and glycopeptide intermediate S. aureus strains in the data set. Differentiation based on the data in broader spectral regions was much less reliable. The results of this study indicate that with proper spectral region selection, FTIR spectroscopy and cluster analysis may provide a simple and accurate means of CNS species identification.

Fourier transform infrared study of lipoxygenase conformation in organic solvent media.

The secondary structure of commercially purified soybean lipoxygenase (EC 1.13.11.12) was investigated in selected monophasic organic solvents, including chloroform, methanol, acetonitrile, hexane, and octane. The Fourier transform infrared (FT-IR) spectra of the enzyme obtained in chloroform, methanol, and acetonitrile showed an absorption band at 1617 cm(-1) indicative of significant protein aggregation, whereas spectra of lipoxygenase in hexane and octane exhibited substantially less aggregate formation. Variable-temperature infrared studies of lipoxygenase in D(2)O show that the predominately alpha-helical structure of the protein undergoes an irreversible transition to intermolecular beta-sheet at and above 65 degrees C. Chemical imaging technology employing an FT-IR spectrometer equipped with an infrared microscope and a focal-plane array detector was used to examine the changes in the secondary structure of lipoxygenase at the water-hexane interface in the presence and absence of substrate. The secondary structure of lipoxygenase at the hexane-water interface was comparable to that of the structure of lipoxygenase in D(2)O after exposure of lipoxygenase solution to hexane.

Heterospectral two-dimensional correlation spectroscopy of mid-infrared and Fourier self-deconvolved near-infrared spectra of sugar solutions.

The mid- and near-infrared (mid-IR and NIR) spectra of aqueous solutions of glucose and fructose, fructose and galactose, and glucose and galactose were recorded and analyzed by heterospectral two-dimensional correlation spectroscopy (H2D-CS) to determine characteristic NIR wavelengths for each sugar. Fourier self-deconvolution (FSD) was applied to the NIR spectra prior to H2D-CS analysis to help resolve the strongly overlapping sugar absorptions. Examination of the H2D-CS data gave characteristic absorption wavelengths for glucose, fructose, and galactose. The wavelengths identified by H2D-CS were then used to develop multiple linear regression (MLR) calibrations for the quantitative analysis of mixtures of the three sugars in solution. This approach gave comparable results to MLR calibrations based on wavelengths selected by examination of the first- and second-derivative spectra of solutions of the individual sugars.