Matija MILANIC1,2, Rok HREN1,3,4, Jost STERGAR1,2, Urban SIMONCIC1,2 1Faculty of Mathematics and Physics, University of Ljubljana, Ljubljana, Republic of Slovenia, 2Jožef Stefan Institute, Ljubljana, Republic of Slovenia, 3Institute of Mathematics, Physics, and Mechanics, Ljubljana, Republic of Slovenia, 4Syreon Research Institute, Budapest, Hungary Physiol Res 2024 Mar 11;73(1):47-56. doi: 10.33549/physiolres.935138. PMID: 38466004 This paper has been retracted on the base of author ́s request.
- MeSH
- Adult MeSH
- Caffeine * administration & dosage MeSH
- Skin * drug effects MeSH
- Humans MeSH
- Young Adult MeSH
- Spectrum Analysis methods MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Young Adult MeSH
- Male MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
The GPCR signalling cascade is a key pathway responsible for the signal transduction of a multitude of physical and chemical stimuli, including light, odorants, neurotransmitters and hormones. Understanding the structural and functional properties of the GPCR cascade requires direct observation of signalling processes in high spatial and temporal resolution, with minimal perturbation to endogenous systems. Optical microscopy and spectroscopy techniques are uniquely suited to this purpose because they excel at multiple spatial and temporal scales and can be used in living objects. Here, we review recent developments in microscopy and spectroscopy technologies which enable new insights into GPCR signalling. We focus on advanced techniques with high spatial and temporal resolution, single-molecule methods, labelling strategies and approaches suitable for endogenous systems and large living objects. This review aims to assist researchers in choosing appropriate microscopy and spectroscopy approaches for a variety of applications in the study of cellular signalling. LINKED ARTICLES: This article is part of a themed issue Complexity of GPCR Modulation and Signaling (ERNST). To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v182.14/issuetoc.
- MeSH
- Humans MeSH
- Microscopy * methods MeSH
- Receptors, G-Protein-Coupled * chemistry metabolism MeSH
- Signal Transduction MeSH
- Spectrum Analysis * methods MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Review MeSH
Laser-induced breakdown spectroscopy (LIBS) is a promising technique for the readout of immunochemical assays utilizing indirect detection of labels (Tag-LIBS), typically based on nanoparticles. We have previously demonstrated that Tag-LIBS immunoassay employing yttrium-based photon-upconversion nanoparticles (UCNPs) can reach sensitivity similar to commonly used enzyme and fluorescence immunoassays. In this study, we report on further increasing the sensitivity of UCNP-based Tag-LIBS immunoassay by employing magnetic microbeads (MBs) as the solid phase in the determination of cancer biomarker prostate-specific antigen. Due to the possibility of analyte preconcentration, MBs enabled achieving a limit of detection (LOD) of 4.0 pg·mL-1, representing two orders of magnitude improvement compared with equivalent microtiter plate-based assay (LOD of 460 pg·mL-1). In addition, utilizing MBs opens up the possibility of an internal standardization of the LIBS readout by employing iron spectral lines, which improves the assay robustness by compensating for LIBS signal fluctuations and bead-bound immunocomplexes lost throughout the washing steps. Finally, the practical applicability of the technique was confirmed by the successful analysis of clinical samples, showing a strong correlation with the standard electrochemiluminescence immunoassay. Overall, MB-based Tag-LIBS was confirmed as a promising immunoassay approach, combining fast readout, multiplexing possibilities, and high sensitivity approaching upconversion luminescence scanning while avoiding the requirement of luminescence properties of labels.
- MeSH
- Immunoassay methods MeSH
- Lasers * MeSH
- Humans MeSH
- Limit of Detection * MeSH
- Microspheres MeSH
- Prostate-Specific Antigen * analysis immunology blood MeSH
- Spectrum Analysis methods MeSH
- Yttrium chemistry radiation effects MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Background: Recent research has linked the spread of microribonucleic acid (miRNA) to numerous disorders, either as a stimulant or an inhibitor. One of these is miRNA-22, which research has connected to oxidative stress and thyroid issues. However, the underlying mechanisms are unknown. This study investigates the expression of miRNA-22 in hypothyroid women and its relationship to the rise in oxidative stress in the patient population.Materials and Methods: 40 women patients with Hypothyroid and 40 in this study, healthy volunteers who served as controls were included. The levels of serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), and thyroid-stimulating hormone (TSH) were measured by sandwich assay, while free triiodothyronine (FT3) and thyroxine (T4) levels were measured competitive binding immunoenzymatic assay. To assess lipid profiles, an automated analyzer was employed. By enzyme-linked immunosorbent assay (ELISA), Interleukin 6 (IL-6) levels were measured. Malondialdehyde (MDA), superoxide dismutase activity (SOD), catalase activity (CAT), and advanced oxidation protein products (AOPPs), and assessed using a colorimetric technique. The quantitative polymerase chain reaction was used to evaluate the expression of serum miRNA-22.Results: Significantly more SOD and CAT activity was identified in patient groups than in the control group (P<0.05), also the patient group's AOPP and MDA concentrations were discovered to significantly outweigh those of the control group. (P< 0.05). IL-6 levels were significantly higher in the patient group than in (P<0.05) the control group. The level of miRNA-22 was higher in the sick group as compared to the control groups (P<0.05).Conclusions: The pathophysiology of oxidative stress brought on by hypothyroidism involves miRNA-22 expression, there is a reciprocal relationship between the increase in gene expression of the miRNA-22 and the increase in oxidative stress, which results in the disease's development.
- MeSH
- Biomarkers blood MeSH
- Chemistry Techniques, Analytical methods instrumentation MeSH
- Adult MeSH
- Hormones blood MeSH
- Hypothyroidism * blood MeSH
- Humans MeSH
- MicroRNAs * blood MeSH
- Oxidative Stress MeSH
- Spectrum Analysis methods instrumentation MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Female MeSH
- Publication type
- Clinical Study MeSH
Correlative imaging of cutaneous tumors provides additional information to the standard histopathologic examination. However, the joint progress in the establishment of analytical techniques, such as Laser-Induced Breakdown Spectroscopy (LIBS) and Laser Ablation Inductively Coupled Plasma Mass Spectrometry (LA-ICP-MS) in clinical practice is still limited. Their combination provides complementary information as it is also shown in our study in terms of major biotic (Ca, Mg, and P) and trace (Cu and Zn) elements. To elucidate changes in the elemental composition in tumors, we have compiled a set of malignant tumors (Squamous Cell Carcinoma, Basal Cell Carcinoma, Malignant Melanoma, and Epithelioid Angiosarcoma), one benign tumor (Pigmented Nevus) and one healthy-skin sample. The data processing was based on a methodological pipeline involving binary image registration and affine transformation. Thus, our paper brings a feasibility study of a practical methodological concept that enables us to compare LIBS and LA-ICP-MS results despite the mutual spatial distortion of original elemental images. Moreover, we also show that LIBS could be a sufficient pre-screening method even for a larger number of samples according to the speed and reproducibility of the analyses. Whereas LA-ICP-MS could serve as a ground truth and reference technique for preselected samples.
- MeSH
- Carcinoma, Basal Cell diagnostic imaging MeSH
- Mass Spectrometry methods MeSH
- Laser Therapy MeSH
- Lasers MeSH
- Humans MeSH
- Melanoma diagnostic imaging pathology MeSH
- Skin Neoplasms * diagnostic imaging pathology MeSH
- Nevus, Pigmented diagnostic imaging MeSH
- Spectrum Analysis methods MeSH
- Carcinoma, Squamous Cell diagnostic imaging pathology MeSH
- Trace Elements analysis MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Diagnostika a verifikace plicních malignit směřuje v poslední době hlavně cestou miniinvazivních technik. V některých případech je verifikace zatížená nedostatečným množstvím odebraného materiálu. Chirurgická verifikace je proto v některých případech nevyhnutelná. Klíč k úspěchu je kombinace vhodně zvolené zobrazovací a diagnostické techniky kombinovaná s erudicí lékaře. I v následujícím článku je místo zvolené verifikace méně tradiční, naštěstí ale s úspěšným diagnostickým závěrem.
x
- MeSH
- Endoscopic Ultrasound-Guided Fine Needle Aspiration * methods MeSH
- Bronchoscopy methods MeSH
- Microscopy, Confocal MeSH
- Middle Aged MeSH
- Humans MeSH
- Neoplasm Metastasis diagnostic imaging diagnosis MeSH
- Adrenal Gland Neoplasms * diagnosis MeSH
- Lung Neoplasms diagnostic imaging MeSH
- Tomography, Optical Coherence MeSH
- Spectrum Analysis methods MeSH
- Check Tag
- Middle Aged MeSH
- Humans MeSH
- Female MeSH
- Publication type
- Case Reports MeSH
The diagnosis of malignant melanoma, often an inconspicuous but highly aggressive tumor, is most commonly done by histological examination, while additional diagnostic methods on the level of elements and molecules are constantly being developed. Several studies confirmed differences in the chemical composition of healthy and tumor tissue. Our study presents the potential of the LIBS (Laser-Induced-Breakdown Spectroscopy) technique as a diagnostic tool in malignant melanoma (MM) based on the quantitative changes in elemental composition in cancerous tissue. Our patient group included 17 samples of various types of malignant melanoma and one sample of healthy skin tissue as a control. To achieve a clear perception of results, we have selected two biogenic elements (calcium and magnesium), which showed a dissimilar distribution in cancerous tissue from its healthy surroundings. Moreover, we observed indications of different concentrations of these elements in different subtypes of malignant melanoma, a hypothesis that requires confirmation in a more extensive sample set. The information provided by the LIBS Imaging method could potentially be helpful not only in the diagnostics of tumor tissue but also be beneficial in broadening the knowledge about the tumor itself.
- MeSH
- Magnesium * analysis MeSH
- Lasers * MeSH
- Humans MeSH
- Melanoma * pathology diagnostic imaging diagnosis chemistry MeSH
- Skin Neoplasms * pathology diagnostic imaging MeSH
- Spectrum Analysis * methods MeSH
- Calcium analysis MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Caffeine is the most widely consumed psychoactive substance worldwide, affecting numerous tissues and organs, with notable impacts on the central nervous system, heart, and blood vessels. The effect of caffeine on vascular smooth muscle cells is an initial transient contraction followed by significant vasodilatation. In this study we investigate the use of diffuse reflectance spectroscopy (DRS) for monitoring of vascular changes in human skin induced by caffeine consumption. DRS spectra were recorded on volar sides of the forearms of eight healthy volunteers at time intervals of 0, 30, 60, 120, and 180 min after consumption of caffeine, while one subject served as a negative control. Analytical diffusion approximation solutions for diffuse reflectance from three-layer structures were used to assess skin composition (e.g. dermal blood volume fraction and oxygen saturation) by fitting these solutions to experimental data. The results demonstrate that cutaneous vasodynamics induced by caffeine consumption can be monitored by DRS, while changes in the control subject not consuming caffeine were insignificant.
- MeSH
- Caffeine * MeSH
- Skin * blood supply MeSH
- Humans MeSH
- Spectrum Analysis methods MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- MeSH
- Densitometry * methods MeSH
- Humans MeSH
- Osteoporosis MeSH
- Spectrum Analysis methods MeSH
- Check Tag
- Humans MeSH
Hepatocelulární karcinom (HCC) je nejčastější primární zhoubný nádor jater, který se ve většině případů rozvíjí v terénu jaterní cirhózy různé etiologie. Jen časná stadia onemocnění jsou indikována k chirurgické, potenciálně kurativní léčbě. Jen přibližně třetina HCC je zachycena v časných stadiích. Screeningovým vyšetřením rizikových skupin pacientů je ultrasonografie jater (USG) v 6měsíčních intervalech. USG má vedle svých známých výhod i značné limitace. Senzitivita USG pro časná stadia HCC se pohybuje pouze okolo 60 %. Vzhledem k tomu a i proto, že se jedná o expert dependentní metodu, existuje naléhává potřeba objektivního biomarkeru HCC. Alfa-fetoprotein je obecně vnímán jako biomarker HCC, nicméně jeho senzitivita a specificita pro účely diagnostiky či dokonce screeningu je nedostatečná. Byly zkoumány i jiné další biomarkery s různými výsledky, ale žádný nedosáhl efektivity USG. Poměrně novým přístupem k této problematice je spektroskopie krevní plazmy, která prokázala svoji účinnost u různých onemocnění. Z pohledu HCC se spektroskopii krevní plazmy věnovalo jen málo studií. Autoři prezentují v přehledu i vlastní práci, kdy spektroskopie krevní plazmy dosáhla senzitivity a specificity v odlišení nemocných s jaterní cirhózou bez HCC a s ním 88 %, respektive 90 %. Přes veškeré snahy nebyl dosud dostatečně spolehlivý a validovaný biomarker HCC použitelný pro účely časné diagnostiky a screeningu identifikován.
Hepatocellular carcinoma (HCC) is the most common primary malignant tumor of the liver, which in most cases develops in the field of liver cirrhosis of various etiologies. Only the early stages of the disease are indicated for surgical, potentially curative treatment. Only about a third of cases of HCC are caught in the early stages. The screening examination of risk groups of patients is liver ultrasonography (USG) at 6-month intervals. Besides its well-known advantages, USG has significant limitations. The sensitivity of USG for the early stages of HCC is only around 60 %. Considering this and also because it is an expert-dependent method, there is an urgent need for an objective HCC biomarker. Alpha-fetoprotein is generally perceived as a biomarker of HCC, however, its sensitivity and specificity for diagnostic or even screening purposes are insufficient. Other biomarkers have been investigated with varying results, but none have reached the effectiveness of USG. A relatively new approach to this problem is blood plasma spectroscopy, which has proven its effectiveness in various diseases. From the perspective of HCC, few studies have focused on blood plasma spectroscopy. In the review, the authors also present their work, where blood plasma spectroscopy achieved a sensitivity and specificity of 88 % and 90 %, respectively, in differentiating patients with liver cirrhosis without and with HCC. Despite all efforts, a sufficiently reliable and validated biomarker of HCC usable for early diagnosis and screening has not yet been identified.
- MeSH
- alpha-Fetoproteins analysis MeSH
- Early Detection of Cancer methods MeSH
- Carcinoma, Hepatocellular * diagnosis MeSH
- Liver Cirrhosis complications MeSH
- Carcinogenesis MeSH
- Plasma MeSH
- Humans MeSH
- Biomarkers, Tumor analysis classification MeSH
- Spectrum Analysis methods MeSH
- Check Tag
- Humans MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
- Review MeSH
