Feasibility of excitation-emission fluorescence spectroscopy in tandem with chemometrics for quantitation of trans-resveratrol in vine-shoot ethanolic extracts.

BACKGROUND: trans-Resveratrol (TR) is a well-known phytochemical compound with important biological properties. It can be recovered from agri-food by-products or wastes, such as vine shoots. Once recovered, its concentration should be measured, possibly in a green, non-destructive, and efficient manner. With these premises, this work aimed to explore the feasibility of excitation-emission fluorescence spectroscopy combined with chemometrics for the analysis of TR in raw extracts obtained from vine shoots. A total of 75 extracts were produced and analyzed by ultra-performance liquid chromatography method with diode array detection (UPLC-DAD) and spectrofluorimetry. Then, the feasibility of two calibration strategies for TR quantitation was assessed - a parallel factor analysis (PARAFAC)-based calibration and the N-way partial least squares (NPLS) regression. RESULTS: The extracts showed variable TR content, the excitation/emission maxima of which were at around 305/390 nm, respectively. The best PARAFAC-based calibration allowed a root mean square error of prediction (RMSEP) of 22.57 mg L-1 , and a relative prediction deviation (RPD) of 2.91 to be obtained but a large number of PARAFAC components should be considered to improve the predictions. The results of the NPLS regression were slightly better, with a RMSEP of 19.47 mg L-1 , and an RPD of 3.33 in the best case. CONCLUSION: Fluorescence could be an alternative analytical technique to measure TR in complex samples. Chemometric tools allowed the identification of the TR signal in the fluorescence landscapes, which could be further used for its non-destructive quantitation. The need for a more accurate criterion for optimal PARAFAC complexity emerged. © 2024 Society of Chemical Industry.

5-Deazaalloxazine as photosensitizer of singlet oxygen and potential redox-sensitive agent.

Flavins are a unique class of compounds that combine the features of singlet oxygen generators and redox-dependent fluorophores. From a broad family of flavin derivatives, deazaalloxazines are significantly underdeveloped from the point of view of photophysical properties. Herein, we report photophysics of 5-deazaalloxazine (1a) in water, acetonitrile, and some other solvents. In particular, triplet excited states of 1a in water and in acetonitrile were investigated using ultraviolet-visible (UV-Vis) transient absorption spectroscopy. The measured triplet lifetimes for 1a were all on the microsecond time scale (≈ 60 µs) in deoxygenated solutions. The quantum yield of S1 → T1 intersystem crossing for 1a in water was 0.43 based on T1 energy transfer from 1a to indicaxanthin (5) acting as acceptor and on comparative actinometric measurements using benzophenone (6). 1a was an efficient photosensitizer for singlet oxygen in aerated solutions, with quantum yields of singlet oxygen in methanol of about 0.76, compared to acetonitrile ~ 0.74, dichloromethane ~ 0.64 and 1,2-dichloroethane ~ 0.54. Significantly lower singlet oxygen quantum yields were obtained in water and deuterated water (Ð¤Δ ~ 0.42 and 0.44, respectively). Human red blood cells (RBC) were used as a cell model to study the antioxidant capacity in vitro and cytotoxic activity of 1a. Fluorescence-lifetime imaging microscopy (FLIM) data were analyzed by fluorescence lifetime parameters and distribution for different parts of the emission spectrum. Comparison of multidimensional fluorescent properties of RBC under physiological-like and oxidative-stress conditions in the presence and absence of 1a suggests its dual activity as probe and singlet-oxygen generator and opens up a pathway for using FLIM to analyze complex intracellular behavior of flavin-like compounds. These new data on structure-property relationship contribute to the body of information required for a rational design of flavin-based tools for future biological and biochemical applications.

Bacterial Metabolites: A Link between Gut Microbiota and Dermatological Diseases.

Dysbiosis has been identified in many dermatological conditions (e.g., psoriasis, atopic dermatitis, systemic lupus erythematosus). One of the ways by which the microbiota affect homeostasis is through microbiota-derived molecules (metabolites). There are three main groups of metabolites: short-chain fatty acids (SCFAs), tryptophan metabolites, and amine derivatives including trimethylamine N-oxide (TMAO). Each group has its own uptake and specific receptors through which these metabolites can exert their systemic function. This review provides up-to-date knowledge about the impact that these groups of gut microbiota metabolites may have in dermatological conditions. Special attention is paid to the effect of microbial metabolites on the immune system, including changes in the profile of the immune cells and cytokine disbalance, which are characteristic of several dermatological diseases, especially psoriasis and atopic dermatitis. Targeting the production of microbiota metabolites may serve as a novel therapeutic approach in several immune-mediated dermatological diseases.

Protective Effects of a Strawberry Ellagitannin-Rich Extract against Pro-Oxidative and Pro-Inflammatory Dysfunctions Induced by a High-Fat Diet in a Rat Model.

Due to the demonstrated intestinal microbial transformation of strawberry ellagitannins (ET) into bioactive metabolites, in the current study on rats, we hypothesised that the dietary addition of a strawberry ET-rich extract (S-ET) to a high-fat diet (HFD) would attenuate disturbances in the redox and lipid status as well as in the inflammatory response. We randomly distributed 48 Wistar rats into six groups and used two-way analysis of variance (ANOVA) to assess the effects of two main factors-diet type (standard and high-fat) and ET dosage (without, low, and 3× higher)-applied to rats for 4 weeks. In relation to the hypothesis, irrespective of the dosage, the dietary application of ET resulted in the desired attenuating effects in rats fed a HFD as manifested by decreased body weight gain, relative mass of the epididymal pad, hepatic fat, oxidized glutathione (GSSG), triglycerides (TG), total cholesterol (TC), and thiobarbituric acid-reactive substances (TBARS) concentrations as well as desired modifications in the blood plasma parameters. These beneficial changes were enhanced by the high dietary addition of ET, which was associated with considerably higher concentrations of ET metabolites in the urine and plasma of rats. The results indicated that S-ET could be effectively used for the prevention and treatment of metabolic disturbances associated with obesity, dyslipidaemia, redox status imbalance, and inflammation.

Non-destructive determination of strawberry fruit and juice quality parameters using ultraviolet, visible, and near-infrared spectroscopy.

BACKGROUND: The development of rapid methods for the determination of the soluble solids content (SSC) and total phenolic content (TPC) in fruit juices is of great interest. Soluble solids content is related to sensory attributes, whereas TPC is related to the antioxidant capacity of juices. The aim of this study was to develop and optimize the calibration models for the prediction of the SSC and TPC of strawberry juices from the spectra of fruit and juices. RESULTS: Near infrared (NIR) spectra were measured for strawberry fruit and ultraviolet (UV), visible (VIS), and NIR spectra were measured for juices. The partial least squares regression models were validated using the test sample set and their predictive ability was evaluated on the basis of determination coefficients (R2 P ) and root mean square error of prediction (RMSEP). For SSC the models with high predictive ability were obtained using spectra of fruit (R2 P = 0.929, RMSEP = 0.46%) or juices (R2 P = 0.979, RMSEP = 0.25%) in the NIR range. The optimal models for TPC were obtained using NIR spectra of fruit (R2 P = 0.834, RMSEP = 130.8 mg GA L-1 ) or UV-VIS-NIR spectra of juices (R2 P = 0.844, RMSEP = 126.7 mg GA L-1 ). CONCLUSION: The results show the potential of spectroscopy for predicting quality parameters of strawberry juices from the juice spectra itself or non-destructively from the fruit spectra. They may contribute to the development of fruit sorting systems to optimize their use in juice production, as well as fast-screening methods for quality control of juices. © 2019 Society of Chemical Industry.

Evaluation of total phenolic content in virgin olive oil using fluorescence excitation-emission spectroscopy coupled with chemometrics.

BACKGROUND: Determination of the total phenolic content (TPC) in olive oils is of great interest, as phenolic compounds affect the health benefits, sensory attributes and oxidative stability of olive oils. The aim of this study was to explore the feasibility of direct front-face fluorescence measurements coupled with chemometrics for developing multivatiate models for discrimination between virgin olive oils with low and high TPC and determination of TPC concentration. RESULTS: Parallel factor analysis and principal component analysis of fluorescence excitation-emission matrices (EEMs) of virgin olive oils revealed different fluorescent properties for samples with low and high TPC. A perfect discrimination of oils with low and high TPC was achieved using partial least squares (PLS) discriminant analysis. The best regression model for the prediction of TPC was based on the PLS analysis of the unfolded entire EEMs (R2  = 0.951, RPD = 4.0). CONCLUSIONS: The results show the potential of fluorescence spectroscopy for direct screening of virgin olive oils for TPC. This may contribute to the development of fast screening methods for TPC assessment, providing an alternative to conventional assays. The procedure is environmentally friendly and fulfils the requirements for green analytical chemistry. © 2018 Society of Chemical Industry.

The effects of rapeseed meal and legume seeds as substitutes for soybean meal on productivity and gastrointestinal function in rabbits.

The aim of this study was to determine the effects of soybean meal (SBM) substitution by a mixture of rapeseed meal (RSM), white lupine seeds (WLS) and pea seeds (PS) on productivity, nutrient digestibility, nitrogen retention and gastrointestinal function in Hyplus rabbits. The Control diet (SBM15) contained 15% SBM, whereas Diet SBM7.5 contained 7.5% SBM, 5% RSM, 4% WLS and 3% PS. In Diet SBM0, SBM was completely replaced by RSM, WLS and PS (10%, 8% and 6%, respectively). A production trial was performed on 90 Hyplus rabbits aged from 35 to 84 d (45 each sex; 953 ± 4.6 g). A digestion and balance trial was conducted on 30 rabbits. Additionally, several parameters of the gastrointestinal tracts from eight animals from each group were analysed, where special attention was paid to the enzymatic activity of microbiota and the short-chain fatty acids concentration in caecum and colon. The experimental diets did not cause significant differences regarding performance parameters evaluated in vivo and post-mortem, and in the nutrient and energy digestibility or nitrogen retention. The observed changes in the enzymatic activity of large gut microbiota, including the selective increase in secretion of glycoside hydrolases by bacterial cells, seem to be responsible for the unchanged growth performance of rabbits fed diets where SBM was substituted by a mixture of RSM, WLS and PS. The obtained results indicate that in rabbit diets SBM may be, partially or completely, successfully replaced by a feed mixture of RSM, WLS and PS.

Preparation and Analysis of Histological Slides of Rat and Mouse Eyeballs to Evaluate the Retina.

A rodent eyeball is a powerful tool for researching the pathomechanisms of many ophthalmic diseases, such as glaucoma, hypertensive retinopathy, and many more. Preclinical experiments enable researchers to examine the efficacy of novel drugs, develop new methods of treatment, or seek new pathomechanisms involved in the disease's onset or progression. A histological examination provides a lot of information necessary to assess the effects of the conducted experiments and can reveal degeneration, tissue remodeling, infiltration, and many other pathologies. In clinical research, there is rarely any chance of obtaining eye tissue suitable for a histological examination, which is why researchers should take advantage of the opportunity offered by the examination of eyeballs from rodents. This manuscript presents a protocol for the histological preparation of rodent eyeballs' sections. The procedure is presented for the eyeballs of mice and rats and has the following steps: (i) harvesting the eyeball, (ii) preserving the eyeball for further analysis, (iii) processing the tissue in paraffin, (iv) preparing slides, (v) staining with hematoxylin and eosin, (vi) assessing the tissue under a light microscope. With the proposed method, the retina can be easily visualized and assessed in detail.

The etiology, diagnostics, and treatment of the spasm of the near reflex - a narrative review.

Physiological adaptation of the eye to the visual perception of near objects consists of the "near triad": convergence, accommodation, and pupil miosis. Normally, these tend to revert when one stops fixating on a near object. Spasm of the near reflex (SNR) is a pathological phenomenon, which manifests itself by the persistence of the above-mentioned adjustments, which prevents the eye from returning to its relaxed state. In this narrative review, we aim to summarize the etiology, diagnostics, treatment, and prevention of SNR. The literature review was performed by searching online databases. The clinical presentation of SNR is diverse; it presents as isolated accommodative spasm more frequently than impairment of all three components of the near triad. Patients usually present with fluctuations in visual acuity, blurred vision, diplopia, and asthenopia. The etiology is not fully understood. Potential causes include neuroanatomic, organic, and psychogenic disorders. The diagnosis is clinical, based on the constellation of symptoms and assessment of the near triad. The diagnostic golden standard is a cycloplegic examination of refraction, preferably using cyclopentolate hydrochloride (1%, 0.5%, or 0.1% solution). The first-line treatment requires the administration of a cycloplegic drug in combination with plus lenses, flipper lenses, optical fogging, or miotics. For secondary cases, causal treatment should be implemented. Prevention of SNR should be based on eliminating modifiable risk factors. We propose including screening for SNR symptoms in every ophthalmic examination, especially among patients with psychogenic or neural disorders, after brain trauma, or young adults spending much time in front of computer screens.

Evaluation of selected parameters of inflammation, coagulation system, and formation of extracellular neutrophil traps (NETs) in the perioperative period in patients undergoing endovascular treatment of thoracoabdominal aneurysm with a branched device (t-Branch).

Extracellular Neutrophils Traps (NETs) and their formation, known as NETosis, have become pivotal in the pathogenesis of aortic aneurysm development. This study investigates the NETosis markers with the assessment of selected parameters of inflammation and coagulation system in patients with thoracoabdominal aortic aneurysms in the pre-and postop period undergoing t-Branch stent-graft implantation. The study included 20 patients with thoracoabdominal aortic aneurysms. Three markers double-stranded DNA (dsDNA), single-stranded DNA (ssDNA), and citrullinated H3 histones (Cit-H3) were tested at three-time points from patients' blood. The parameters of NETosis, inflammation, and coagulation system were examined in the preoperative period (within 24 h before surgery) and in the postoperative period (on the 3rd and 5th postoperative day). Free-circulating DNA (cfDNA) was isolated from the blood using the MagMAXTM Cell-Free DNA Extraction Kit. Double-stranded DNA (dsDNA) and single-stranded DNA (ssDNA) were then quantified using the Qubit dsDNA HS Assay Kit and the Qubit ssDNA Assay Kit. Cit-H3 concentration was determined by enzyme immunoassay ELISA (Cayman). The results revealed the significance of NETs secretion in response to the complex processes after stent-graft implantation. All NET markers increased shortly after surgery, with histones being the first to return to preoperative levels. The lack of normalization of dsDNA and ssDNA levels to preoperative levels by the last postoperative blood collection demonstrates NETs reorganization. The increase in the number of neutrophils was not related to the expansion of postoperative NETosis. The study reveals a new marker of NETosis, ssDNA, that has not been studied so far. The implantation of a stent graft in a patient with TAAA triggers an inflammatory response manifested by an increase in inflammatory parameters. One of the hallmarks of inflammation is the activation of neutrophil extracellular traps.

Tetramethylalloxazines as efficient singlet oxygen photosensitizers and potential redox-sensitive agents.

Tetramethylalloxazines (TMeAll) have been found to have a high quantum yield of singlet oxygen generation when used as photosensitizers. Their electronic structure and transition energies (S0 → Si, S0 → Ti, T1 → Ti) were calculated using DFT and TD-DFT methods and compared to experimental absorption spectra. Generally, TMeAll display an energy diagram similar to other derivatives belonging to the alloxazine class of compounds, namely π,π* transitions are accompanied by closely located n,π* transitions. Photophysical data such as quantum yields of fluorescence, fluorescence lifetimes, and nonradiative rate constants were also studied in methanol (MeOH), acetonitrile (ACN), and 1,2-dichloroethane (DCE). The transient absorption spectra were also analyzed. To assess cytotoxicity of new compounds, a hemolytic assay was performed using human red blood cells (RBC) in vitro. Subsequently, fluorescence lifetime imaging experiments (FLIM) were performed on RBC under physiological and oxidative stress conditions alone or in the presence of TMeAll allowing for pinpointing changes caused by those compounds on the intracellular environment of these cells.

Effect of dietary supplementation with unprocessed and ethanol-extracted apple pomaces on caecal fermentation, antioxidant and blood biomarkers in rats.

The present 4-week study on growing Wistar rats was aimed at assessing the potential advantages of dietary supplementation with apple pomace containing both fibre and polyphenols, which enables nutritional exploitation of the physiological traits of both compounds. A total of twenty-four rats, assigned to the control (C), group fed with a diet supplemented with 14% of processed apple pomace (A) and group fed with a diet supplemented with 15% unprocessed apple pomace (AP) groups, were fed the following iso-fibrous diets: control, ethanol-extracted and unprocessed apple pomaces (low and high level of dietary polyphenols, i.e. 0·002 and 0·018%, respectively). To measure the animal response, parameters describing the caecal fermentation, antioxidative status and lipoprotein profile of rats were assessed. Both dietary apple pomaces were found to significantly (P≤0·05) decrease caecal pH and ammonia concentration, microbial ß-glucuronidase activity as well as to increase caecal SCFA concentration in comparison to the control diet. The unprocessed pomace did not suppress caecal fermentation. Unlike the extracted one, the dietary apple pomace rich in polyphenols significantly (P≤0·05 v. C group) increased erythrocyte superoxide dismutase activity and serum antioxidant capacity of lipid-soluble substances and significantly (P≤0·05 v. C group) decreased amounts of thiobarbituric acid-reactive substances in liver tissue. Moreover, the 4-week administration of the AP diet to rats evoked a significant decrease in serum glucose concentration (P≤0·05 v. C and A groups). In conclusion, the results demonstrated that the polyphenol-rich fibre complexes from apple pomace exerted positive effects on gastrointestinal physiology and antioxidant status of rats.

Acid-base equilibriums of lumichrome and its 1-methyl, 3-methyl, and 1,3-dimethyl derivatives.

Lumichrome photophysical properties at different pH were characterized by UV-vis spectroscopy and steady-state and time-resolved fluorescence techniques, in four forms of protonation/deprotonation: neutral form, two monoanions, and dianion. The excited-state lifetimes of these forms of lumichrome were measured and discussed. The results were compared to those obtained for similar forms of alloxazine and/or isoalloxazine, and also to those of 1-methyl- and 3-methyllumichrome and 1,3-dimethyllumichrome. The absorption, emission, and synchronous spectra of lumichrome, 1-methyl- and 3-methyllumichrome, and 1,3-dimethyllumichrome at different pH were measured and used in discussion of fluorescence of neutral and deprotonated forms of lumichrome. The analysis of steady-state and time-resolved spectra and the DFT calculations both predict that the N(1) monoanion and the N(1,3) dianion of lumichrome have predominantly isoalloxazinic structures. Additionally, we confirmed that neutral lumichrome exists in its alloxazinic form only, in both the ground and the excited state. We also confirmed the existence and the alloxazinic structure of a second N(3) monoanion. The estimated values of pK(a) = 8.2 are for the equilibrium between neutral lumichrome and alloxazinic and isoalloxazinic monoanions, with proton dissociation from N(1)-H and N(3)-H groups proceeding at the almost the same pH, while the second value pK(a) = 11.4 refers to the formation of the isoalloxazinic dianion in the ground state.

Color and luminescence stability of selected dental materials in vitro.

PURPOSE: To study luminescence, reflectance, and color stability of dental composites and ceramics. MATERIALS AND METHODS: IPS e.max, IPS Classic, Gradia, and Sinfony materials were tested, both unpolished (as-cast) and polished specimens. Coffee, tea, red wine, and distilled water (control) were used as staining drinks. Disk-shaped specimens were soaked in the staining drinks for up to 5 days. Color was measured by a colorimeter. Fluorescence was recorded using a spectrofluorometer, in the front-face geometry. Time-resolved fluorescence spectra were recorded using a laser nanosecond spectrofluorometer. RESULTS: The exposure of the examined dental materials to staining drinks caused changes in color of the composites and ceramics, with the polished specimens exhibiting significantly lower color changes as compared to unpolished specimens. Composites exhibited lower color stability as compared to ceramic materials. Water also caused perceptible color changes in most materials. The materials tested demonstrated significantly different initial luminescence intensities. Upon exposure to staining drinks, luminescence became weaker by up to 40%, dependent on the drink and the material. Time-resolved luminescence spectra exhibited some red shift of the emission band at longer times, with the lifetimes in the range of tens of nanoseconds. CONCLUSIONS: Unpolished specimens with a more developed surface have lower color stability. Specimens stored in water develop some changes in their visual appearance. The presently proposed methods are effective in evaluating the luminescence of dental materials. Luminescence needs to be tested in addition to color, as the two characteristics are uncorrelated. It is important to further improve the color and luminescence stability of dental materials.

Characterization and Classification of Direct and Commercial Strawberry Beverages Using Absorbance-Transmission and Fluorescence Excitation-Emission Matrix Technique.

The subject of this study was to characterize the absorption and fluorescence spectra of various types of strawberry beverages and to test the possibility of distinguishing between direct juices and pasteurized commercial products on the basis of their spectral properties. An absorbance and transmission excitation-emission matrix (A-TEEMTM) technique was used for the acquisition of spectra. The obtained spectra were analyzed using chemometric methods. The principal component analysis (PCA) revealed differences in both the absorption spectra and excitation-emission matrices (EEMs) of two groups of juices. The parallel factor analysis (PARAFAC) enabled the extraction and characterization of excitation and emission profiles and the relative contribution of four fluorescent components of juices, which were related to various groups of polyphenols and nonenzymatic browning products. Partial least squares-discriminant analysis (PLS-DA) models enabled 100% correct class assignment using the absorption spectra in the visible region, unfolded EEMs, and set of emission spectra with excitation at wavelengths of 275, 305, and 365 nm. The analysis of variable importance in projection (VIP) suggested that the polyphenols and nonenzymatic browning products may contribute significantly to the differentiation of commercial and direct juices. The results of the research may contribute to the development of fast methods to test the quality and authenticity of direct and processed strawberry juices.

Effect of the dietary polyphenolic fraction of chicory root, peel, seed and leaf extracts on caecal fermentation and blood parameters in rats fed diets containing prebiotic fructans.

The aim of this 28 d experiment was to examine the physiological response of growing rats to a dietary combination of prebiotic chicory fructans, with polyphenols originating from different parts of the chicory plant, i.e. roots, root peels, seeds and leaves. A total of forty rats were assigned to groups fed the following diets characterised by a similar content of oligofructose and inulin: control, with 10 % of a root extract (a low level of dietary polyphenols, 0·05 %), with 6·5 % of a root peel extract (a medium level of dietary polyphenols, 0·107 %), with a combination of 8 % of a peel extract and 0·8 % of a seed extract (a high level of dietary polyphenols, 0·208 %) and with 2·5 % of a leaf extract (a medium level of dietary polyphenols, 0·106 %, with chicoric acid constituting half of them). Chicory seeds are the richest source of polyphenols, especially abundant in dicaffeoylquinic acids. When applied as a dietary supplement, the mixture of monocaffeoylquinic and dicaffeoylquinic acids, from the extracts made of roots, root peels and seeds, elicited more favourable changes in parameters of the antioxidative status of the body and in the activity of bacterial ß-glucuronidase in the faeces and caecal digesta. In turn, the extract from chicory leaves, containing considerable quantities of chicoric acid and polyphenolic glycosides, apart from chlorogenic acids, also triggered desirable changes in the lipid profile of the blood serum. The high concentration of polyphenols in the extracts examined enables their application as dietary supplements to be administered in low doses.

Strawberry phenolic extracts effectively mitigated metabolic disturbances associated with high-fat ingestion in rats depending on the ellagitannin polymerization degree.

In the present experiment it was hypothesised that dietary strawberry ellagitannin-rich extracts would mitigate negative consequences associated with consumption. Therefore, two extracts rich in dimeric (D-ET) or monomeric (M-ET) ellagitannins (ETs) were added to a standard or high-fat diet fed to rats for four weeks. The D-ET-rich extract contained 82.3% polyphenols, and the M-ET/D-ET ratio was 40 : 60, while the M-ET-rich extract contained 88.0% and 96 : 4, respectively. The experimental feeding with high-fat diets containing extracts resulted in beneficial mitigating effects in the lipid profile, redox status of the rat's liver and blood plasma. According to the accepted hypothesis, the obtained results pointed at increased desired hepatic and plasma modifications when the extract was rich in M-ET, as indicated by favourable changes in the hepatic fat content, GSH and GSSG concentrations and GSH/GSSG ratio as well as blood plasma FRAP, ACL, HDL-cholesterol, and atherogenic coefficient values. These changes were partly connected to the fact that M-ET was more prone vs. D-ET to intestinal microbial conversion into respective metabolites. The urinary daily excretion of ET metabolites and their blood plasma concentrations were higher in rats fed with M-ET vs. D-ET-rich diets. To conclude, the metabolic action of the M-ET-rich extract in the normalization of high-fat-induced disturbances was more pronounced.

A new heterotropic vascularized model of total urinary bladder transplantation in a rat model.

This study developed a new procedure of urinary bladder transplantation on a rat model (n = 40). Heterotopic urinary bladder transplantation (n = 10) in the right groin vessels was performed. Direct urinary bladder examination, microangiography, histological analysis, and India ink injection were performed to evaluate the proposed method's functionality. Observation time was four weeks. One week after the procedure, the graft survival rate was 80%, two urinary bladders were lost due to anastomosis failure. The rest of the grafts survived two weeks without any complications. Lack of transitional epithelium or smooth muscle layer loss and lack of inflammatory process development were observed. This study was performed in order to obtain the necessary knowledge about urinary bladder transplantation. The proposed technique offers a new approach to the existing orthotropic models.

Application of multidimensional and conventional fluorescence techniques for classification of beverages originating from various berry fruit.

The objectives of this study were to characterize fluorescence of beverages from berry fruit, including chokeberry, blackcurrant, raspberry and strawberry, and to develop classification models based on different types of fluorescence spectra to identify beverages depending on the fruit species. Total fluorescence spectra (excitation-emission matrices, EEMs) and total synchronous fluorescence spectra (TSFS) were recorded for a series of commercial berry fruit beverages. An analysis of EEMs using parallel factor analysis (PARAFAC) revealed four components characterized by the excitation/emission maxima at 275/326, 319/410, 414/600, and 360/460 nm, respectively. Based on the spectral profiles, these components were assigned to various groups of phenolic compounds. Partial least squares discriminant analysis was used to develop the classification models. The analysis was performed on PARAFAC scores, unfolded EEMs (uEEMs), unfolded TSFS (uTSFS), and additionally on conventional emission spectra (EMS) measured at particular excitation wavelengths and single synchronous fluorescence spectra (SFS). The classification models with the same average classification error of 4.86% were obtained for the analysis of both the entire uEEMs and uTSFS. Among models based on the individual spectra, the lowest error of 4.42% was obtained for SFS measured at Δλ = 40 nm, and an error of 7.64% was obtained for EMS measured at the excitation wavelength of 360 nm. The classification model based on the PARAFAC scores had the highest error of 15.27%. The present results show good potential of fluorescence as rapid and reagent-free tool for authenticity evaluation of berry beverages.

Monitoring Virgin Olive Oil Shelf-Life by Fluorescence Spectroscopy and Sensory Characteristics: A Multidimensional Study Carried Out under Simulated Market Conditions.

The control of virgin olive oil (VOO) freshness requires new tools that reflect the diverse chemical changes that take place during the market period. Fluorescence spectroscopy is one of the techniques that has been suggested for controlling virgin olive oil (VOO) freshness during its shelf-life. However, a complete interpretation of fluorescence spectra requires analyzing multiple parameters (chemical, physical-chemical, and sensory) to evaluate the pace of fluorescence spectral changes under moderate conditions with respect to other changes impacting on VOO quality. In this work, four VOOs were analyzed every month with excitation-emission fluorescence spectra. The same samples were characterized with the concentration of fluorophores (phenols, tocopherols, chlorophyll pigments), physical-chemical parameters (peroxide value, K232, K270, free acidity), and sensory attributes (medians of defects and of the fruity attribute). From the six components extracted with parallel factor analysis (PARAFAC), two components were assigned to chlorophyll pigments and those assigned to tocopherols, phenols, and oxidation products were selected for their ability to discriminate between fresh and aged oils. Thus, the component assigned to oxidation products correlated with K270 in the range 0.80-0.93, while the component assigned to tocopherols-phenols correlated with the fruity attribute in the range 0.52-0.90. The sensory analysis of the samples revealed that the changes of these PARAFAC components occurred at the same time as, or even before, the changes of the sensory characteristics.