Morphological and tissue characterization of the medicinal fungus Hericium coralloides by a structural and molecular imaging platform

In this study the potential of new imaging techniques such as Magnetic Resonance Imaging (MRI), Matrix-Assisted Laser Desorption/Ionization (MALDI) profiling mass spectrometry ("MALDI Profiling") and Fourier Transform Infrared (FTIR) spectroscopic imaging was evaluated to study morphological and molecular patterns of the potential medicinal fungus Hericium coralloides. For interpretation, the MALDI profiling, FTIR imaging and MRI results were correlated with histological information gained from Scanning Electron Microscopy (SEM) and Light Microscopy (LM). Additionally we tested several evaluation processes and optimized the methodology for use of complex FTIR images to monitor molecular patterns. It is demonstrated that the combination of these spectroscopic methods enables to gain a more distinct picture concerning morphology and distribution of active ingredients. We were able to obtain high quality FTIR imaging and MALDI-profiling results and to distinguish different tissue types with their chemical ingredients. Beside this, we have created a 3-D reconstruction of a mature Hericium basidioma, based on the MRI dataset: analyses allowed, for the first time, a realistic approximation of the "evolutionary effectiveness" of this bizarrely formed basidioma type, concerning the investment of sterile tissue and its reproductive output (production of basidiospores).     

Screening and evaluation of antioxidants from lees by micro-injector systems combined with a fluorescent probe,N-borylbenzyloxycarbonyl-3,7-dihydroxyphenoxazine,in living Drosophila

A HPLC-UV-FLD system and a micro-injector imaging system were used in the study of antioxidants from natural products.     

Diffusion of water–acetic acid mixtures in epoxy

The solubility and diffusion of water–acetic acid solutions in epoxy resins has been studied by nuclear magnetic resonance imaging (MRI) and spectroscopy (NMR), Fourier transform infrared (FTIR) spectroscopy, and light microscopy (LM). These techniques revealed the progression of a sharp diffusion front at a rate proportional to the square root of time. Both the swelling and rate of diffusion are dependent on acid dilution. At higher acid concentrations, hydrogen bonds were present, which has been interpreted as formation of acid dimers. The results indicate that molecular interactions play a major role in controlling the solubility and diffusion processes. The observation that voids can be filled up with penetrant strongly support the interpretation that molecular interactions rather than concentration gradient are responsible for these effects. © 1999 John Wiley & Sons, Inc. J Polym Sci B: Polym Phys 37: 3328–3336, 1999     

Fourier transform infrared imaging analysis in discrimination studies of squamous cell carcinoma

Oral squamous cell carcinoma (OSCC) of the oral cavity and oropharynx represents more than 95% of all malignant neoplasms in the oral cavity. Histomorphological evaluation of this cancer type is invasive and remains a time consuming and subjective technique. Therefore, novel approaches for histological recognition are necessary to identify malignancy at an early stage. Fourier transform infrared (FTIR) imaging has become an essential tool for the detection and characterization of the molecular components of biological processes, such as those responsible for the dynamic properties of tumor progression. FTIR imaging is a modern analytical technique enabling molecular imaging of a complex biological sample and is based on the absorption of IR radiation by vibrational transitions in covalent bonds. One major advantage of this technique is the acquisition of local molecular expression profiles, while maintaining the topographic integrity of the tissue and avoiding time-consuming extraction, purification, and separation steps. With this imaging technique, it is possible to obtain unique images of the spatial distribution of proteins, lipids, carbohydrates, cholesterols, nucleic acids, phospholipids, and small molecules with high spatial resolution. Analysis and visualization of FTIR imaging datasets are challenging and the use of chemometric tools is crucial in order to take advantage of the full measurement. Therefore, methodologies for this task based on the novel developed algorithm for multivariate image analysis (MIA) are often necessary. In the present study, FTIR imaging and data analysis methods were combined to optimize the tissue measurement mode after deparaffinization and subsequent data evaluation (univariate analysis and MIAs). We demonstrate that it is possible to collect excellent IR spectra from formalin-fixed paraffin-embedded (FFPE) tissue microarrays (TMAs) of OSCC tissue sections employing an optimised analytical protocol. The correlation of FTIR imaging to the morphological tissue features obtained by histological staining of the sections demonstrated that many histomorphological tissue patterns can be visualized in the colour images. The different algorithms used for MIAs of FTIR imaging data dramatically increased the information content of the IR images from squamous cell tissue sections. These findings indicate that intra-operative and surgical specimens of squamous cell carcinoma tissue can be characterized by FTIR imaging.     

Differentiation of individual human mesenchymal stem cells probed by FTIR microscopic imaging

Objective of this study is the novel application of Fourier transform infrared (FTIR) microscopic imaging to identify the differentiation state of individual human mesenchymal stem cells with or without osteogenic stimulation. IR spectra of several hundred single cells with lateral resolution of 5-10 microm were recorded using a FTIR imaging spectrometer coupled to a microscope with a focal plane array detector. A classification model based on linear discriminant analysis was trained to distinguish four cell types by their IR spectroscopic fingerprint. Without stimulation two cell types dominated, showing low or high levels of glycogen accumulation at the cell periphery. After stimulation, the protein composition in the cells changed and some cells started expressing calcium phosphate salts such as octacalciumphosphate, a precursor of the bone constituent hydroxyapatite. Few cells were identified which remained in their non-stimulated state. This study demonstrated for the first time that FTIR microscopic imaging can probe stem cell differentiation at the single cell level rapidly, non-destructively and with minimal preparation.     

Facile Synthesis of PEGylated Tungsten-based Nanoprobes for Gastric Computed Tomography Imaging

With the development of science and technology, PEGylated nanoprobes have attracted a great deal of interest in the molecular imaging area of modern medicine. These nanoprobes play an important role especially in diagnosis of gastric diseases. The transition metal oxides as gastric contrast agents based on PEGylated WO_3?x nanoprobes were designed in this work. The nanoprobes were developed via a high-temperature solvent thermal method, and then modified with polyethylene glycol (PEG) to improve stability and dispersivity of nanoprobes in water. The characterizations of PEGylated WO_3?x nanoprobes included scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FTIR) spectroscopy analysis etc. Moreover, in vivo toxicological studies were carried out with histopathology. Otherwise, as a contrast agent, multi-slice computed tomography (MDCT) imaging were illustrated both in vitro and in vivo assessment. The results indicated that PEGylated WO_3?x nanoprobes had great potential for gastric MDCT imaging in detection of gastric diseases.     

Unique evolution of spatial and dynamic heterogeneities on the glass transition behavior of PVPh/PEO blends

Glass transition behavior of hydrogen bonded polymer blends of poly(vinyl phenol)(PVPh) and poly(ethylene oxide)(PEO) is systematically investigated using normal differential scanning calorimetry(DSC) and recently developed multifrequency temperature-modulated DSC(TOPEM),in combination with Fourier transform infrared spectroscopy(FTIR) and nuclear magnetic resonance(NMR) techniques,focusing on the effect of the PEO molecular weight on the spatial and dynamic heterogeneity.It is found,for the first time,that both the glass transition temperature(T_g) and activity energy(E_a) of the blends strongly depend on PEO molecular weight,and a common turning point,which separates the rapid and slow increasing regions,can be found.The interchain hydrogen bonding interactions,both determined by FTIR measurements and obtained from the Kwei equation,decrease with increasing PEO molecular weight,indicating a decrease of the componential miscibility.A series of parameters related to the microscopic spatial and dynamic heterogeneity,such as the activity energy, fragility,nonexponential factor and the size of cooperatively rearranging regions,are calculated from frequency dependency complex heat capacity measured using TOPEM.It is found that each of these parameters monotonically changes with increasing the PEO molecular weight during the glass transition process,demonstrating that hydrogen bonding interaction is the key factor in controlling the spatial and dynamic heterogeneity,thus the glass transition.NMR relaxation results reveal the existence of obvious phase separation large than 5 nm,implying that the cooperatively rearranging regions should be closely related to the interphase region between the two components.The above obtained origin and evolution of spatial and dynamic heterogeneity provide a new insight into the glass transition behavior of polymer blends.     

Tautomerism in liquid 1,2,3-triazole: a combined energy-dispersive X-ray diffraction, molecular dynamics, and FTIR study

In this work, we report a multitechnique (energy-dispersive X-ray diffraction, computational methods, and FTIR spectroscopy) study of the tautomeric equilibrium of 1,2,3-triazole, one of the few small nitrogen-containing eterocycles liquid at room temperature. The T-2H form (C _2v symmetry) is found to be strongly favored in gas and solid phases, whereas the neat liquid gives diffraction patterns that can be interpreted satisfactorily with the structure functions calculated from some molecular dynamics results for both T-2H and T-1H tautomers, although the T-2H form gives a slightly better agreement.     

Systemic chemical characterization of Lemna minor by UHPLC-Q-Exactive Orbitrap MS coupled with parallel reaction monitoring

Lemna minor L. (LM) has been used for measles opacity, rubella itching, edema, and oliguria, and the main active ingredients were flavonoids, namely, apigenin, apigenin-7-O-glucoside, and luteolin-7-O-glucoside. However, few systematic analyses of their constituents have been performed; thus, it was necessary to establish a fast and efficient method to identify the chemical composition of LM. In this study, the UHPLC-Q-Exactive Orbitrap mass spectrometry coupled with parallel reaction monitoring was established. Finally, a total of 112 constituents, including 30 dipeptides, 28 nucleosides, 11 amino acids, 10 organic acids, 10 flavonoids, and 23 other compounds, were identified by MS, diagnostic fragment ions, and retention time. One hundred one of those chemicals were first found in LM, which was very beneficial for the further development and utilization of nutriments and the medicinal use of LM.     

Drying dissipative structures of lightly cross-linked poly(2-vinyl pyridine) cationic gel spheres stabilized with poly(ethylene glycol) in the deionized aqueous suspension

Drying dissipative patterns of deionized and colloidal crystal-state suspensions of the cationic gel spheres of lightly cross-linked poly(2-vinyl pyridine) stabilized with poly(ethylene glycol) were observed on a cover glass, a watch glass, and a Petri glass dish. Convectional patterns were recognized with the naked eyes. The broad rings were observed in the drying pattern and their size and width decreased as gel concentration decreased. Formation of the monodispersed agglomerated particles and their ordered arrays were observed. This work clarified the formation of the drying microscopic structures of (a) ordered rings, (b) flickering ordered spoke-lines, (c) net structure, and (d) lattice-like ordered structures of the agglomerated particles. The ordering of the agglomerated particles of the cationic gel spheres is similar to that of the anionic thermo-sensitive gel spheres of poly(N-isopropyl acrylamide). The role of the electrical double layers around the agglomerated particles and the interaction of the particles with the substrates during dryness are important for the ordering. The microscopic drying patterns of gel spheres were different from those of linear-type polymers and also from typical colloidal hard spheres, though the macroscopic patterns such as broad ring formation at the edges were similar to each other. The addition of sodium chloride shifted the microscopic patterns from lattice to net structures.     

Raman mapping and FTIR imaging of lung tissue: congenital cystic adenomatoid malformation

Congenital cystic adenomatoid malformations (CCAMs) are benign masses of non-functional lung tissue developing from an overgrowth of the terminal bronchioles with subsequent suppressing of alveolar growth. For the first time CCAMs are studied by Raman mapping and Fourier transform infrared (FTIR) imaging. Both vibrational spectroscopic methods are able to analyze the biochemical composition of tissues and their pathological changes at the molecular level. Cryosections were prepared on calcium fluoride substrates from CCAMs and from normal lung tissue of two infant patients who underwent surgery. Raman maps were collected at a step size of 100 microm in order to assess the whole tissue section and at a smaller step size of 10 microm in order to resolve details in selected areas. FTIR images were collected in the macroscopic and microscopic modes. Data sets were segmented by cluster analysis and the mean spectra of each cluster were compared. At low lateral resolution a lower red blood cell content and higher lipid content were found in CCAMs than in normal lung tissue. At higher lateral resolution, accumulations of lipids and glycogen were identified in CCAMs. The lipid aggregates contain a high concentration of phosphatidylcholine. It is discussed how the combined application of Raman mapping and FTIR imaging might improve the differential diagnosis of lung malformations and how both modalities might be applied to other bioanalytical and biomedical problems in the future.     

Catalyst optimization for enhanced propylene formation in the methanol-to-olefins reaction

ZSM-5 zeolites possessing different chemical compositions, acidities and crystal sizes, were synthesized and characterized by XRD, SEM and FTIR. Those as-prepared acid catalysts were tested in the conversion of methanol into light olefins at 673 K under atmospheric pressure. Propylene-to-ethylene ratios above 5 were achieved over ZSM-5 zeolite catalyst prepared via the fluoride route. This promising methanol-to-propylene (MTP) catalyst was designed both at the molecular scale, exhibiting low Brønsted acid site density, and at the microscopic level since large crystals (25 μm) with few defects were obtained. Aiming in a possible industrial application, the synthesis medium has to be seriously modified, thus avoiding harmful hydrofluoric acid use. Hydrochloric and phosphoric acids were therefore chosen as alternatives and resulted in the same ZSM-5 crystal morphologies and similar catalytic performance.     

Review of FTIR microspectroscopy applications to investigate biochemical changes in C. elegans

Caenorhabditis elegans nematode has emerged as a model organism paving the ways for multidisciplinary research in biomedical, environmental toxicology, aging, metabolism, obesity, and drug discovery. The wide range of applications of this model organism are attributed to C. elegans’ unique features: C. elegans are inexpensive, easy to grow and maintain in a laboratory, has a short lifespan, and has a small body size. With this increased interest, the need for analytical techniques to assess the biochemical information on intact worms continues to grow. Fourier Transform Infrared (FTIR) microspectroscopy is considered as a powerful technique that can be used to determine the chemical structure and composition of various materials, including biological samples. Furthermore, the development of focal plane array detectors has made this technique attractive to study complex biological systems such as whole nematodes. This review focuses on the use of FTIR microspectroscopy to study C. elegans. The first published work on the use of FTIR microspectroscopy to study a complex whole animal was reported in 2004. Since then, very few other studies were carried out. The objective of this review is to summarize work conducted to date using FTIR microspectroscopy to study nematodes and to discuss the information that can be gained by using this technique. This could allow scientists to add this technique to the arsenal of techniques already in use for C. elegans studies.     

Screening of Potential Anti-Thrombotic Ingredients from Salvia miltiorrhiza in Zebrafish and by Molecular Docking

Background: Danshen (DS), the dry root of Salvia miltiorrhiza Bge., has been used in traditional Chinese medicine (TCM) for many years to promote blood circulation and to inhibit thrombosis. However, the active ingredients responsible for the anti-thrombotic effect and the underlying mechanisms are yet to be fully elucidated. Methods: Molecular docking was used to predict the active ingredients in DS and their potential targets by calculating the scores of docking between DS ingredients and thrombosis-related proteins. Then, a chemical-induced zebrafish thrombosis model was applied to confirm their anti-thrombotic effects. Result: The molecular docking results indicated that compared to the control ligand, higher docking scores were observed for several compounds in DS, among which salvianolic acid B (SAB), lithospermic acid (LA), rosmarinic acid (MA), and luteolin-7-O-β-d-glucoside (LG) could attenuate zebrafish caudal vein thrombosis and recover the decrease in heart red blood cells (RBCs) in a dose-dependent manner. Conclusions: Our study showed that it is possible to screen the potential active components in natural products by combining the molecular docking method and zebrafish in vivo model.     

Trends in the Use of Botanicals in Anti-Aging Cosmetics

Botanical ingredients have been used for thousands of years in skincare for their convenience as well as the diversity and abundance in compounds with biological activity. Among these, polyphenols and especially flavonoids have gained increasing prominence due to their antioxidant and anti-inflammatory properties. In this study, the most used botanical preparations in anti-aging products marketed in 2011 were determined. The analysis was repeated in 2018 for new and reformulated products. The scientific evidence for their application as active ingredients in anti-aging cosmetics and their flavonoid content was also compiled by searching in online scientific databases. Overall, in 2018, there was a noticeable increase in the use of botanical preparations in anti-aging cosmetics. However, the top three botanical species in both years were Vitis vinifera, Butyrospermum parkii, and Glycine soja, which is consistent with the greater amount of scientific evidence supporting their efficacy. Regarding the function of botanical preparations, there is a clear preference for DNA-protecting ingredients. The most prevalent flavonoids were flavan-3-ols, proanthocyanidins, and anthocyanins. This study provided an updated overview of the market trends regarding the use of botanicals in anti-aging products and documented the state of the art of scientific evidence for the most used plants.     

Thermoanalytical study of some anti-inflammatory analgesic agents

Thermogravimetry (TG), differential thermal analysis (DTA), differential scanning calorimetry (DSC) as well as X-ray diffraction powder (DRX) patterns and Fourier transformed infrared spectroscopy (FTIR) were used to study ketoprofen, ibuprofen, and naproxen. The chemical or physical properties of the studied compounds were established and when possible by X-ray powder diffractometry and/or infrared spectroscopy were used. In this investigation, quantum chemical approach was used to determine the molecular structures using Becke three-parameter hybrid method and the Lee–Yang–Par (LYP) correlation functional. The performed molecular calculations in this work were done using the Gaussian 03 routine. Theoretical calculations help in interpretations of FTIR spectra supplying structural and physicochemical parameters.     

Py-GC/MS, GC/MS and FTIR investigations on LATE Roman-Egyptian adhesives from opus sectile: New insights into ancient recipes and technologies

An analytical protocol based on optical microscopy, Fourier transforms infrared spectroscopy (FTIR), analytical pyrolysis in the presence of hexamethyldisilazane followed by gas chromatographic/mass spectrometric analysis (Py-GC/MS) and gas chromatography/mass spectrometry after alkaline hydrolysis, solvent extraction and trimethylsilylation (GC/MS) was used in the chemical characterisation of the original adhesives used to fix monochrome and mosaic glass and stone plaques coming from the Late Roman archaeological site of Antinoopolis (Egypt).FTIR analysis demonstrated the presence of calcite fragments, and Py-GC/MS and GC/MS analyses provided detailed molecular compositions, highlighting the presence of a wide range of compound classes including diterpenoid acids, tricyclic abietanes with a high degree of aromatisation, mid- and long-chain monocarboxylic fatty acids, mono- and di-hydroxy acids, α,ω-dicaboxylic fatty acids, n-alkanols, and n-alkanes. Characteristic biomarkers and their distribution patterns indicated the presence of pine pitch in all the adhesives, which in some cases was admixed with beeswax and brassicaceae seed oil.The results provided new insights into the complex recipes used by artisans in ancient Egypt in the production of adhesives and in the sophisticated manufacture of opus sectile decorations.     

Characterisation and Classification of Foodborne Bacteria Using Reflectance FTIR Microscopic Imaging

This work investigates the application of reflectance Fourier transform infrared (FTIR) microscopic imaging for rapid, and non-invasive detection and classification between Bacillus subtilis and Escherichia coli cell suspensions dried onto metallic substrates (stainless steel (STS) and aluminium (Al) slides) in the optical density (OD) concentration range of 0.001 to 10. Results showed that reflectance FTIR of samples with OD lower than 0.1 did not present an acceptable spectral signal to enable classification. Two modelling strategies were devised to evaluate model performance, transferability and consistency among concentration levels. Modelling strategy 1 involves training the model with half of the sample set, consisting of all concentrations, and applying it to the remaining half. Using this approach, for the STS substrate, the best model was achieved using support vector machine (SVM) classification, providing an accuracy of 96% and Matthews correlation coefficient (MCC) of 0.93 for the independent test set. For the Al substrate, the best SVM model produced an accuracy and MCC of 91% and 0.82, respectively. Furthermore, the aforementioned best model built from one substrate was transferred to predict the bacterial samples deposited on the other substrate. Results revealed an acceptable predictive ability when transferring the STS model to samples on Al (accuracy = 82%). However, the Al model could not be adapted to bacterial samples deposited on STS (accuracy = 57%). For modelling strategy 2, models were developed using one concentration level and tested on the other concentrations for each substrate. Results proved that models built from samples with moderate (1 OD) concentration can be adapted to other concentrations with good model generalization. Prediction maps revealed the heterogeneous distribution of biomolecules due to the coffee ring effect. This work demonstrated the feasibility of applying FTIR to characterise spectroscopic fingerprints of dry bacterial cells on substrates of relevance for food processing.     

Inclusional association as studied by the drying dissipative structure. Part 3. Drying pattern of aqueous mixtures of β-cyclodextrin and n-alkyltrimethylammonium bromide

Macroscopic and microscopic drying patterns were observed on a cover glass and a watch glass during the course of dryness of aqueous mixtures of β-cyclodextrin (βCD) with n-alkyltrimethylammonium bromide (alkyl: n-decyl, n-dodecyl, n-tetradecyl, and n-hexadecyl). Macroscopic patterns of the broad rings and the spoke lines appeared. The inclusional association constants of βCD with the cationic surfactants were determined from the bent points in the curves of the drying broad-ring size vs. surfactant concentration when a finite amount of cyclodextrin presents. The short and thick rod-like microscopic drying patterns were observed for the inclusional complexes.     

Fourier transform infrared spectroscopy supported by multivariate statistics in compatibility study of atenolol with excipients

The objective of this study was to use Fourier transform infrared spectroscopy (FTIR) and multivariate statistics to investigate compatibility/incompatibility of atenolol as a representative of active pharmaceutical ingredients and excipients, such as β-cyclodextrin, methylcellulose, starch and chitosan, when used in solid dosage formulations. Two-component physical mixtures consisting of atenolol and selected excipients were studied by FTIR spectroscopy and two methods of multivariate statistical analysis – principal component analysis (PCA) and cluster analysis (CA), which were used as a supplementary tool for interpretation of the FTIR spectra. Taking into account variability explained by the first two principal components, the results of PCA were visualized in the form of a bi-dimensional scatterplot. A lack of interaction was confirmed by two distinct clusters created by both atenolol and a particular excipient with their mixtures. In the case of CA, lack of interaction between both ingredients was also indicated by two large clusters at a level of 33 or 66% of the maximum distance. The results of the investigations show that with the exception of β-cyclodextrin, the remaining excipients are compatible with atenolol. These findings were confirmed by complementary methods, such as differential scanning calorimetry, thermogravimetry and X-ray powder diffraction.