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Vibrational characterization of granulosa cells from patients affected by unilateral ovarian endometriosis: New insights from infrared and Raman microspectroscopy
摘要： Endometriosis is a chronic gynaecological disease characterised by the presence of endometrial cells in extra-uterine regions. One of the main factors impacting on the fertility of women affected by endometriosis is the poor oocyte quality. Granulosa Cells (GCs) regulate oocyte development and maintain the appropriate microenvironment for the acquisition of its competence; hence, the dysregulation of these functions in GCs can lead to severe cellular damages also in oocytes. In this study, luteinized GCs samples were separately collected from both ovaries of women affected by Unilateral Ovarian Endometriosis and analyzed by infrared and Raman microspectroscopy. The spectral data were compared with those of GCs from women with diagnosis of tubal, idiopathic or male infertility (taken as control group). The coupling of these two spectroscopic techniques sheds new light on the alteration induced by this pathology on GCs metabolism and biochemical composition. In fact, the study revealed similar biochemical modifications in GCs from both ovaries of women affected by unilateral ovarian endometriosis, such as the alteration of the protein pattern, the induction of oxidative stress mechanisms, and the deregulation of lipid and carbohydrate metabolisms. These evidences suggest that unilateral endometriosis impairs the overall ovarian functions, causing alterations not only in the ovary with endometriotic lesions but also in the contralateral “healthy” one.
关键词： Unilateral Ovarian Endometriosis,Multivariate analysis,FTIR microspectroscopy,Raman microspectroscopy
更新于2025-09-23 15:23:52
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Carotenoids co-localize with hydroxyapatite, cholesterol, and other lipids in calcified stenotic aortic valves. Ex vivo Raman maps compared to histological patterns
摘要： Unlike its application for atherosclerotic plaque analysis, Raman microspectroscopy was sporadically used to check the sole nature of bioapatite deposits in stenotic aortic valves, neglecting the involvement of accumulated lipids/lipoproteins in the calcific process. Here, Raman microspectroscopy was employed for examination of stenotic aortic valve leaflets to add information on nature and distribution of accumulated lipids and their correlation with mineralization in the light of its potential precocious diagnostic use. Cryosections from surgically explanted stenotic aortic valves (n=4) were studied matching Raman maps against specific histological patterns. Raman maps revealed the presence of phospholipids/triglycerides and cholesterol, which showed spatial overlapping with one another and Raman-identified hydroxyapatite. Moreover, the Raman patterns correlated with those displayed by both von-Kossa-calcium- and Nile-blue-stained serial cryosections. Raman analysis also provided the first identification of carotenoids, which co-localized with the identified lipid moieties. Additional fit concerned the distribution of collagen and elastin. The good correlation of Raman maps with high-affinity staining patterns proved that Raman microspectroscopy is a reliable tool in evaluating calcification degree, alteration/displacement of extracellular matrix components, and accumulation rate of different lipid forms in calcified heart valves. In addition, the novel identification of carotenoids supports the concept that valve stenosis is an atherosclerosis-like valve lesion, consistently with their previous Raman microspectroscopical identification inside atherosclerotic plaques.
关键词： lipids,Valve calcification,Raman microspectroscopy,stenosis,carotenoids
更新于2025-09-23 15:22:29
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Determination of the microplastics emission in the effluent of a municipal waste water treatment plant using Raman microspectroscopy
摘要： Samples from the secondary clarifier effluent of a waste water treatment plant (serving 98500 inhabitants) were analyzed to determine the microplastics (MP) emission. The samples were collected using a stainless steel centrifugal pump and filtered through a 10 mm stainless steel cartridge filter. Microplastics particles (MPPs) and microplastics fibers (MPFs) were recovered by chemical and physical sample purification. To remove natural organic matter, the samples were first subjected to oxidative treatment with H2O2 and NaClO. Inorganic materials were subsequently removed by density separation in ZnCl2 (r = 1.9 g/cm3) using a centrifuge. Special centrifuge tubes were developed for this purpose. Sample analysis was performed on a Si filter by Raman micro-spectroscopy. Particles with a diameter (dp) ≥ 10 mm were analyzed. The results were differentiated by dry and wet weather samples. On average, 5900 MPPs m?3 were identified in the effluent on wet weather days compared to 3000 MPPs m?3 on dry weather days. Most of the MPPs detected were in the 30 mm < dp < 100 mm size range. The MPFs ranged between 100 mm and 1000 mm in length. While most of the MPFs were of PET origin, the MPPs consisted mainly of PET, PP, PE and PS.
关键词： Raman microspectroscopy,Oxidative treatment,Si filter,Centrifugation,Microplastics emission,WWTP effluent
更新于2025-09-23 15:22:29
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Spectral features of nuclear DNA in human sperm assessed by Raman Microspectroscopy: Effects of UV-irradiation and hydration
摘要： Raman Microspectroscopy represents an innovative tool for the assessment of sperm biochemical features otherwise undetectable by routine semen analysis. Previously, it was shown that induced DNA damage can be detected in smeared sperm by this technique. This novel readout may be of value for clinical settings especially if it can be transferred to living cells. Yet, starting with living sperms this study was carried-out using a variety of conditions to disclose the Raman features of sperm nuclei under different hydration conditions and UV exposure. Human sperm were immobilized and Raman spectra were obtained from individual sperm as repeated measurements. To create conditions with controlled DNA damage, sperm samples were exposed to ultraviolet light. Several media were used to evaluate their effect on Raman spectra in aqueous conditions. To substantiate differences between the experimental conditions, the spectra were analyzed by Principal Component Analysis. We observed that spectra of sperm nuclei obtained in different solutions showed a qualitatively unchanged spectral pattern showing the principal signals related to DNA. Evaluating the effect of ultraviolet light generated the finding that spectra representing DNA damage were only observed in dry conditions but not in aqueous medium. Thus, Raman microspectroscopy was successfully applied for sperm analysis in different conditions, among them in live spermatozoa in aqueous solution during the initial measurement, revealing the principle use of this technique. However, implementation of Raman spectroscopy as a technique for clinical sperm analysis and selection may be especially relevant when DNA evaluation can be established using live sperm.
关键词： hydration,DNA damage,sperm,UV-irradiation,Raman Microspectroscopy
更新于2025-09-23 15:21:21