•Raman spectroscopy on nuclear and membrane regions of single human neuroblastoma cells.•Measurements are performed after irradiation by clinically relevant X-ray doses.•The acquired spectra are ...analyzed by PCA and interval-PCA methods.•The most relevant effects are evidenced in the spectra from the cell nucleus region.•Potential markers of an apoptotic process could be singled out for some X- ray doses.
Raman micro-spectroscopy was performed in vitro on nuclear and membrane regions of single SH-SY5Y human neuroblastoma cells after irradiation by graded X-ray doses (2, 4, 6, 8 Gy). The acquired spectra were analyzed by principal component analysis (PCA) and interval-PCA (i-PCA) methods. Biochemical changes occurring in the different regions of single cells as a consequence of the radiation exposure were observed in cells fixed immediately after the irradiation. The most relevant effects arose from the analysis of the spectra from the cell nucleus region. The observed changes were discussed in terms of the modifications in the cell cycle, resulting in an increase in the DNA-related signal, a protein rearrangement and changes in lipid and carbohydrates profiles within the nucleus. Potential markers of an apoptotic process in cell population irradiated with 6 and 8-Gy X-ray doses could have been singled out. No significant effects were found in spectra from cells fixed 24 h after the irradiation, thus suggesting the occurrence of repairing processes of the X-ray induced damage.
•Cellular acute heat shock response based on Fourier transform-IR spectroscopy.•Early cellular response to heat shock induces rearrangements of cellular components.•Cytoplasmic heat shock protein ...genes are essential to counteract acute heat shock.•Oxidative stress is not involved in early cellular response to heat shock.•Activation of early defence mechanisms may be decisive for cellular thermotolerance.
Heat stress is a significant challenge in dairy cattle herds, affecting milk production and quality, and generating important changes at the cellular level. Most in vitro research on heat shock (HS) effects on dairy cow mammary cells was focused on medium-long-term effects. In recent years, Fourier transform-infrared (FT-IR) micro-spectroscopy has been increasingly used to study the effects of several external stresses on different cell lines, down to the level of single cellular components, such as DNA/RNA, lipids, and proteins. In this study, the possible changes at the biochemical and molecular level induced by acute (30 min-2 h) HS in bovine mammary epithelial (BME-UV1) cells were investigated. The cells were exposed to different temperatures, thermoneutral (TN, 37 °C) and HS (42 °C), and FT-IR spectra were acquired to analyse the effects of HS on biochemical characteristics of BME-UV1 cellular components (proteins, lipids, and DNA/RNA). Moreover, cell viability assay, reactive oxygen species production, and mRNA expression of heat shock proteins (HSPA1A, HSP90AA1, GRP78, GRP94) and antioxidant genes (SOD1, SOD2) by RT-qPCR were also analysed. The FT-IR results showed a change already at 30 min of HS exposure, in the content of long-chain fatty acids, which probably acted as a response to a modification of membrane fluidity in HS cells compared with TN cells. After 2 h of HS exposure, modification of DNA/RNA activity and accumulation of aggregated proteins was highlighted in HS cells. The gene expression analyses showed the overexpression of HSPA1A and HSP90AA1 starting from 30 min up to 2 h in HS cells compared with TN cells. At 2 h of HS exposure, also the overexpression of GRP94 was observed in HS cells. Acute HS did not affect cell viability, reactive oxygen species level, and SOD1 and SOD2 gene expression of BME-UV1 cells. According to the results obtained, cells initiate early defence mechanisms in case of acute HS and probably this efficient response capacity may be decisive for tolerance to heat stress of dairy cattle.
Olive oil is the main fat source of the Mediterranean diet. This seasoning ingredient is highly appreciated for its unique taste, functional properties, and positive impact on human health. The ...determination of chemical composition is a demanding task in order to fully characterize this precious food product, ensure its quality, and prevent fraudulent practices. Among innovative techniques proposed for the oil analysis, surface-enhanced Raman spectroscopy (SERS) can be an extremely useful tool for olive oil characterization. In this frame, we have investigated five noncommercial olive oils produced in different parts of South Italy by using a commercial Raman microspectroscopy apparatus and home-made signal-enhancing SERS substrates. A wavelet-based data analysis has allowed us to efficiently remove the background and the noise from the acquired spectra. The analysis of these SERS spectra has enabled the quantification of the relative contents of carotene, oleic acid, and phenols. These relative contents differ in the examined samples. In addition, SERS response in the lipid region has indicated differences in the relative abundance of saturated fatty acids. The present results confirm the validity of the SERS technique as a rapid, nondestructive, and reliable analytical technique for identifying olive oil bioactive components.
Olive oil is a major fat source of the Mediterranean diet. For its unique functional and technological properties, olive oil is highly appreciated all over the world. Very sensitive techniques are ...currently required to determine chemical composition, to evaluate olive oil authenticity and to quantify vegetable adulterants or degradation compounds. A class of techniques that can be particularly interesting in olive oil characterization is represented by those based on light scattering. These techniques can provide important information on physical properties, conservation state and possible adulteration without complicate or time expensive procedures. Among these, static and dynamic light scattering, diffuse wave spectroscopy, different kinds of Raman spectroscopy are the most used. In this short review, basic concepts about the experimental aspects of these techniques are presented together with some of the most generally used data analysis procedures. Some selected examples of the most interesting applications of these techniques are also proposed.
► The potential of Raman spectroscopy for glucose quantitative determination was investigated. ► Micro-Raman spectra in the 600–1600cm−1 region of four sport drinks were recorded. ► A multivariate ...analysis based on the interval Partial Least Square approach was used. ► Glucose concentration results were in good agreement with the values given by a biochemical assay.
The potential of Raman spectroscopy with excitation in the visible as a tool for quantitative determination of single components in food industry products was investigated by focusing the attention on glucose content in commercial sport drinks. At this aim, micro-Raman spectra in the 600–1600cm−1 wavenumber shift region of four sport drinks were recorded, showing well defined and separated vibrational fingerprints of the various contained sugars (glucose, fructose and sucrose). By profiting of the spectral separation of some peculiar peaks, glucose content was quantified by using a multivariate statistical analysis based on the interval Partial Least Square (iPLS) approach. The iPLS model needed for data analysis procedure was built by using glucose aqueous solutions at known sugar concentrations as calibration data. This model was then applied to sport drink spectra and gave predicted glucose concentrations in good agreement with the values obtained by using a biochemical assay. These results represent a significant step towards the development of a fast and simple method for the on-line glucose quantification in products of food and beverage industry.
Aloe barbadensis Miller, known as Aloe Vera, requires limited irrigation depending on the capacity of the soil to retain humidity, since it is a CAM species and thus naturally adapted to conditions ...of dryness and high temperatures. Therefore, we postulated that plants of Aloe Vera plants under conditions of water deficit should improve their water use efficiency (WUE) by performing osmotic adjustment (OA) with a temporal correlation between WUE and OA. The objective of the investigation was to determine the effect of water restriction on the WUE and OA of
A. barbadensis under different water treatments. 18-month old Aloe Vera plants were cultivated in pots with a soil substrate that was a mixture of equal parts of sand and organic matter with 18% of FC and 9% of permanent wilting point. To determine the effects of the soil humidity on plant WUE and OA, four treatments were arranged in a complete random design with four repetitions; these were 100%, 75%, 50% and 25% of FC, which correspond to an evatranspiration of 11.4, 9.6, 4.0 and 1.7
L per plant, respectively. The water treatments were maintained by frequent irrigation. The following variables were determined: dry matter, leaf water potential, relative water content (RWC), amount of gel produced, sap flow, proline content, soluble and total sugars and oligo and polyfructans. Aloe Vera increased WUE with increasing water deficit; the sap flow rate decreased with water restrictions, and the plants performed osmotic adjustment by increasing the synthesis of proline, soluble and total sugars as well as the amounts of oligo and polyfructans, mainly polymers of β-(2
→
6) kestotriose, changing from the inulin type to the neofructan type. The plants most and less irrigated (100% and 25% of FC) were the groups with lowest WUE. The plants irrigated with 75% of FC presented the best WUE in terms of dry mass and amount of gel produced by a litre of supplied water.
An optical model for Raman response of thin metal films has been developed, taking into account attenuation effects and multi-reflections occurring at film edges. Film thickness and surface ...morphology of nanometer thin NbN films have been inferred in the framework of this model using micro-Raman measurements. Results have been compared with those obtained by means of spectroscopic ellipsometry. The absolute value of the thickness is determined with a precision better than 20% on nanometric scale allowing us to control surface morphology with high accuracy also on large areas.
•A micro-Raman spectroscopy method for evaluating film thickness is presented.•Raman signal attenuation and multi-reflections effects in thin films were studied.•Superconductor NbN thin films on MgO substrates have been fabricated and studied.•Thin NbN films have been investigated by micro-Raman spectroscopy and ellipsometry.
To evaluate whether the use of inhaled nitric oxide (iNO)200 improves respiratory function.
This retrospective cohort study used data from pregnant patients hospitalized with severe bilateral ...coronavirus disease 2019 (COVID-19) pneumonia at four teaching hospitals between March 2020 and December 2021. Two cohorts were identified: 1) those receiving standard of care alone (SoC cohort) and 2) those receiving iNO200 for 30 minutes twice daily in addition to standard of care alone (iNO200 cohort). Inhaled nitric oxide, as a novel therapy, was offered only at one hospital. The prespecified primary outcome was days free from any oxygen supplementation at 28 days postadmission. Secondary outcomes were hospital length of stay, rate of intubation, and intensive care unit (ICU) length of stay. The multivariable-adjusted regression analyses accounted for age, body mass index, gestational age, use of steroids, remdesivir, and the study center.
Seventy-one pregnant patients were hospitalized for severe bilateral COVID-19 pneumonia: 51 in the SoC cohort and 20 in the iNO200 cohort. Patients receiving iNO200 had more oxygen supplementation-free days (iNO200: median interquartile range, 24 23-26 days vs standard of care alone: 22 14-24 days, P=.01) compared with patients in the SoC cohort. In the multivariable-adjusted analyses, iNO200 was associated with 63.2% (95% CI 36.2-95.4%; P<.001) more days free from oxygen supplementation, 59.7% (95% CI 56.0-63.2%; P<.001) shorter ICU length of stay, and 63.6% (95% CI 55.1-70.8%; P<.001) shorter hospital length of stay. No iNO200-related adverse events were reported.
In pregnant patients with severe bilateral COVID-19 pneumonia, iNO200 was associated with a reduced need for oxygen supplementation and shorter hospital stay.
Glucose oxidase (GOD) immobilized into sol–gel matrices was studied by using Micro-Attenuated Total Reflection Fourier Transform Infrared (micro-ATR FT-IR) spectroscopy in order to characterize ...enzyme distribution and secondary structure in systems with valuable potentialities in amperometric and optical biosensing. Spectra were acquired in the 4000–600cm−1 frequency region and the analysis of specific fingerprints in the FT-IR spectra evidenced that the enzyme was actually immobilized in the matrix. The enzyme spatial distribution was obtained by examining the amide I and amide II band region of spectra from defined sample positions. The deconvolution of the amide I band in terms of lorentzian functions provided information on the secondary structure of the immobilized GOD. By this approach a macroscopic preservation of GOD activity upon immobilization was evidenced along with the existence of some matrix sites with locally inactivated GOD. To our knowledge this is the first example of point-by-point characterization of conformational changes of immobilized enzyme by means of micro-ATR infrared spectroscopy, thus confirming that this technique can be usefully employed for a non- or minimally-invasive detailed micro-characterization of catalytic supports in order to improve their functionality.
► Sol–gel immobilized GOD has been investigated by means of FT-IR microspectroscopy. ► Secondary structure of immobilized GOD has been obtained at defined spatial positions. ► An overall preservation of the GOD activity upon immobilization has been shown. ► Some matrix sites where there is locally inactivated GOD have been evidenced. ► Enzyme distribution and activity of catalytic supports have been well-characterized.
The of stingless bees creation is an agricultural activity that has grown in recent years. In the Brazilian Northeast, Melipona subnitida is one of the most important species in the Semiarid region. ...Among the products, wax is a substance produced by worker bees and is the main material used in the construction of laying discs and food pots. The present work aims to study the preference and non-preference of Jandaíra bees to different types of wax, their internal behaviors and the flight activity of these bees. The experiments were conducted at the Federal Institute of Paraíba, Sousa campus, in October 2023. Four Jandaíra bee colonies were used, kept in boxes, a northeastern model, with a glass inner side, allowing observations inside the boxes. Circular containers containing different types of wax were placed in each box. After 12:00 o’clock the containers containing the least and most preferred waxes were inverted. Bees were counted in each of the containers every thirty minutes. Additionally, the entry and exit of bees were counted (for a period of 10 minutes every 30 minutes). The data were subjected to the Q-square test using the R Core (2003) software. Uruçú beeswax was the most preferred over the others. In the study of flight activity, peaks of bee departures were observed in the late morning and early afternoon. Peaks were observed at 08:20, 11:20 and 15:20. It can be concluded that: Uruçu beeswax was the most preferred among those studied, in the morning and afternoon, followed by Africanized beeswax. During flight activity, peaks in departures and arrivals can be seen, varying depending on the time of day and between boxes.
PDF
