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17 pages, 7769 KiB

Open AccessArticle

A Novel Approach Using Reduced Graphene Oxide for the Detection of ALP and RUNX2 Osteogenic Biomarkers

by Elena Alina Chiticaru and Mariana Ioniță

Curr. Issues Mol. Biol. 2024, 46(5), 4489-4505; https://doi.org/10.3390/cimb46050272 - 8 May 2024

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Abstract

In this work, we propose a new technique involving the modification of commercial screen-printed carbon electrodes with electrochemically reduced graphene oxide to serve as the starting point of a future electrochemical biosensor for the detection of two osteogenic biomarkers: alkaline phosphatase (ALP) and [...] Read more.

In this work, we propose a new technique involving the modification of commercial screen-printed carbon electrodes with electrochemically reduced graphene oxide to serve as the starting point of a future electrochemical biosensor for the detection of two osteogenic biomarkers: alkaline phosphatase (ALP) and Runt-related transcription factor 2 (RUNX2). The electrodes were characterized after each modification by cyclic voltammetry and electrochemical impedance spectroscopy, showing the appropriate electrochemical characteristics for each modification type. The results obtained from scanning electron microscopy, Raman spectroscopy, X-ray photoelectron spectroscopy, and contact angle measurements are well correlated with each other, demonstrating the successful modification of the electrodes with graphene oxide and its subsequent reduction. The bioreceptors were immobilized on the electrodes by physical adsorption, which was confirmed by electrochemical methods, structural characterization, and contact angle measurements. Finally, the functionalized electrodes were incubated with the specific target analytes and the detection relied on monitoring the electrochemical changes occurring after the hybridization process. Our results indicated that the pilot platform has the ability to detect the two biomarkers up to 1 nM, with increased sensitivity observed for RUNX2, suggesting that after further optimizations, it has a high potential to be employed as a future biosensor. Full article

(This article belongs to the Special Issue Molecular Imaging of Cells and Tissues)

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12 pages, 2270 KiB

Open AccessArticle

Effects of Letrozole Treatment and Vitamin C Supplementation on Morphology, Endoplasmic Reticulum Stress, Programmed Cell Death, and Oxidative Stress in the Small Intestine of Adult Male Rats

by Anna Pilutin, Sylwia Rzeszotek, Aleksandra Wilk, Klaudia Klimaszewska, Julia Łukasiewicz, Rufaro Lynnette Mafuta, Thanushan Nagendran, Rupia Ndambara and Barbara Wiszniewska

Curr. Issues Mol. Biol. 2024, 46(3), 1943-1954; https://doi.org/10.3390/cimb46030127 - 1 Mar 2024

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Abstract

Estrogens are hormones that play an important role in the digestive tract, including in men. Letrozole is an inhibitor of cytochrome P450 aromatase, an enzyme converting androgens to estrogens. The use of letrozole may cause oxidative stress and endoplasmic reticulum stress in the [...] Read more.

Estrogens are hormones that play an important role in the digestive tract, including in men. Letrozole is an inhibitor of cytochrome P450 aromatase, an enzyme converting androgens to estrogens. The use of letrozole may cause oxidative stress and endoplasmic reticulum stress in the cells. Factors modulating cellular stress may include vitamin C. The purpose of this study was to examine whether letrozole and/or vitamin C supplementation can affect the morphology of the small intestine, the parameters of endoplasmic reticulum stress, programmed cell death markers, and oxidative damage. Three-month-old male rats were divided into four groups and treated with the following: (I) CTRL—water; (II) CTRL+C—L-ascorbic acid; (III) LET—letrozole; and (IV) LET+C—letrozole + L-ascorbic acid. The morphometrical measurements included epithelial thickness, crypt and lumen area, crypt perimeter, nuclei number in the crypt, and the cell size of crypts. The expression levels of PERK, caspase-3, and catalase were determined. Significant differences in the morphometrical measurements and immunoexpression were observed. This may indicate that chronic treatment with letrozole can affect morphology and induce ER stress, oxidative stress, and programmed cell death in the epithelial cells of the small intestine of adult male rats. Vitamin C supplementation exerts an effect on some parameters of the molecular processes. Full article

(This article belongs to the Special Issue Molecular Imaging of Cells and Tissues)

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13 pages, 2490 KiB

Open AccessArticle

The Role of Nicotinic Receptors on Ca²⁺ Signaling in Bovine Chromaffin Cells

by Amparo Gil, Virginia González-Vélez, Luis Miguel Gutiérrez and José Villanueva

Curr. Issues Mol. Biol. 2024, 46(1), 808-820; https://doi.org/10.3390/cimb46010052 - 17 Jan 2024

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Abstract

Chromaffin cells have been used as a physiological model to understand neurosecretion in mammals for many years. Nicotinic receptors located in the cells’ membrane are stimulated by acetylcholine, and they participate in the exocytosis of chromaffin granules, releasing catecholamines in response to stress. [...] Read more.

Chromaffin cells have been used as a physiological model to understand neurosecretion in mammals for many years. Nicotinic receptors located in the cells’ membrane are stimulated by acetylcholine, and they participate in the exocytosis of chromaffin granules, releasing catecholamines in response to stress. In this work, we discuss how the participation of nicotinic receptors and the localization of active zones in the borders of the cytoskeleton can generate local calcium signals leading to secretion. We use a computational model of a cytoskeleton cage to simulate Ca²⁺ levels in response to voltage and acetylcholine pulses. We find that nicotinic receptors are able to enhance the differences between local and average calcium values, as well as the heterogeneous distributions around the active zones, producing a non-linear, highly localized Ca²⁺ entry that, although consisting of a few ions, is able to improve secretion responses in chromaffin cells. Our findings emphasize the intricate interplay among nicotinic receptors, the cytoskeleton, and active zones within chromaffin cells as an example of Ca²⁺-dependent neurosecretion in mammals. Full article

(This article belongs to the Special Issue Molecular Imaging of Cells and Tissues)

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9 pages, 4766 KiB

Open AccessCommunication

Comparison of Retinal Metabolic Activity and Structural Development between rd10 Mice and Normal Mice Using Multiphoton Fluorescence Lifetime Imaging Microscopy

by Erin Su, Niranjana Kesavamoorthy, Jason A. Junge, Mengmei Zheng, Cheryl Mae Craft and Hossein Ameri

Curr. Issues Mol. Biol. 2024, 46(1), 612-620; https://doi.org/10.3390/cimb46010039 - 6 Jan 2024

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Abstract

Fluorescence lifetime imaging microscopy (FLIM) is a technique that analyzes the metabolic state of tissues based on the spatial distribution of fluorescence lifetimes of certain interacting molecules. We used multiphoton FLIM to study the metabolic state of developing C57BL6/J and rd10 retinas based [...] Read more.

Fluorescence lifetime imaging microscopy (FLIM) is a technique that analyzes the metabolic state of tissues based on the spatial distribution of fluorescence lifetimes of certain interacting molecules. We used multiphoton FLIM to study the metabolic state of developing C57BL6/J and rd10 retinas based on the fluorescence lifetimes of free versus bound nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate (NAD(P)H), with free NAD(P)H percentages suggesting increased glycolysis and bound NAD(P)H percentages indicating oxidative phosphorylation. The mice were sacrificed and enucleated at various time points throughout their first 3 months of life. The isolated eyecups were fixed, sectioned using a polyacrylamide gel embedding technique, and then analyzed with FLIM. The results suggested that in both C57BL6/J mice and rd10 mice, oxidative phosphorylation initially decreased and then increased, plateauing over time. This trend, however, was accelerated in rd10 mice, with its turning point occurring at p10 versus the p30 turning point in C57BL6/J mice. There was also a noticeable difference in oxidative phosphorylation rates between the outer and inner retinas in both strains, with greater oxidative phosphorylation present in the latter. A greater understanding of rd10 and WT metabolic changes during retinal development may provide deeper insights into retinal degeneration and facilitate the development of future treatments. Full article

(This article belongs to the Special Issue Molecular Imaging of Cells and Tissues)

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24 pages, 3435 KiB

Open AccessArticle

Off the Beaten Path in Oncology: Active Brown Adipose Tissue by Virtue of Molecular Imaging

by Wael Jalloul, Mihaela Moscalu, Roxana Moscalu, Despina Jalloul, Irena Cristina Grierosu, Teodor Ionescu, Cati Raluca Stolniceanu, Vlad Ghizdovat, Veronica Mocanu, Radu Iliescu, Ioana Pavaleanu and Cipriana Stefanescu

Curr. Issues Mol. Biol. 2023, 45(10), 7891-7914; https://doi.org/10.3390/cimb45100499 - 28 Sep 2023

Cited by 1 | Viewed by 1681

Abstract

Brown Adipose Tissue (BAT) is considered beneficial in diabetes and obesity, but it can also have negative effects such as its implication in tumours’ pathogenesis and the development of Cancer-induced Cachexia. Since ¹⁸F-FDG PET/CT is a common molecular imaging modality used in [...] Read more.

Brown Adipose Tissue (BAT) is considered beneficial in diabetes and obesity, but it can also have negative effects such as its implication in tumours’ pathogenesis and the development of Cancer-induced Cachexia. Since ¹⁸F-FDG PET/CT is a common molecular imaging modality used in cancer assessment, we aim to study the ¹⁸F-FDG BAT biodistribution in oncological patients and look for possible correlations between BAT activity and different malignancies as well as the patient’s weight status. After analysing the total number of oncological ¹⁸F-FDG PET/CT scans between 2017 and 2021, we selected patients with active BAT. Based on their BMI, the selected patients were divided into nonobese (NO) vs. overweight and obese (OOB). OOB SUV_maxlean body mass(LBM) had the highest mean values in supraclavicular, latero-cervical, and paravertebral vs. mediastinal and latero-thoracic localisations in NO. BMI was positively correlated with latero-cervical and supraclavicular SUV_max(LBM) but negatively correlated with latero-thoracic and abdominal SUV_max(LBM). Considering the age of the patients, SUV_max(LBM) decreases in the latero-cervical, paravertebral, and abdominal regions. In addition, the males presented lower SUV_max(LBM) values. SUV_max(LBM) was not affected by the treatment strategy or the oncological diagnosis. To conclude, it is mandatory to take into consideration the BAT particularities and effects on weight status in order to optimise the clinical management of oncological patients. Full article

(This article belongs to the Special Issue Molecular Imaging of Cells and Tissues)

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11 pages, 1707 KiB

Open AccessArticle

Flow Cytometry Detection of Anthracycline-Treated Breast Cancer Cells: An Optimized Protocol

by Giulia Catitti, Simone De Fabritiis, Davide Brocco, Pasquale Simeone, Domenico De Bellis, Simone Vespa, Serena Veschi, Laura De Lellis, Nicola Tinari, Fabio Verginelli, Marco Marchisio, Alessandro Cama, Antonia Patruno and Paola Lanuti

Curr. Issues Mol. Biol. 2023, 45(1), 164-174; https://doi.org/10.3390/cimb45010013 - 28 Dec 2022

Cited by 7 | Viewed by 2653

Abstract

The use of anthracycline derivatives was approved for the treatment of a broad spectrum of human tumors (i.e., breast cancer). The need to test these drugs on cancer models has pushed the basic research to apply many types of in vitro assays, and, [...] Read more.

The use of anthracycline derivatives was approved for the treatment of a broad spectrum of human tumors (i.e., breast cancer). The need to test these drugs on cancer models has pushed the basic research to apply many types of in vitro assays, and, among them, the study of anthracycline-induced apoptosis was mainly based on the application of flow cytometry protocols. However, the chemical structure of anthracycline derivatives gives them a strong autofluorescence effect that must be considered when flow cytometry is used. Unfortunately, the guidelines on the analysis of anthracycline effects through flow cytometry are lacking. Therefore, in this study, we optimized the flow cytometry detection of doxorubicin and epirubicin-treated breast cancer cells. Their autofluorescence was assessed both by using conventional and imaging flow cytometry; we found that all the channels excited by the 488 nm laser were affected. Anthracycline-induced apoptosis was then measured via flow cytometry using the optimized setting. Consequently, we established a set of recommendations that enable the development of optimized flow cytometry settings when the in vitro assays of anthracycline effects are analyzed, with the final aim to reveal a new perspective on the use of those in vitro tests for the further implementation of precision medicine strategies in cancer. Full article

(This article belongs to the Special Issue Molecular Imaging of Cells and Tissues)

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Review

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14 pages, 991 KiB

Open AccessReview

MALDI Imaging Mass Spectrometry of High-Grade Gliomas: A Review of Recent Progress and Future Perspective

by Alen Rončević, Nenad Koruga, Anamarija Soldo Koruga, Željko Debeljak, Robert Rončević, Tajana Turk, Domagoj Kretić, Tatjana Rotim, Zdravka Krivdić Dupan, Damir Troha, Marija Perić and Tihana Šimundić

Curr. Issues Mol. Biol. 2023, 45(2), 838-851; https://doi.org/10.3390/cimb45020055 - 18 Jan 2023

Cited by 5 | Viewed by 3262

Abstract

Glioblastoma (GBM) is the most common malignancy of the brain with a relatively short median survival and high mortality. Advanced age, high socioeconomic status, exposure to ionizing radiation, and other factors have been correlated with an increased incidence of GBM, while female sex [...] Read more.

Glioblastoma (GBM) is the most common malignancy of the brain with a relatively short median survival and high mortality. Advanced age, high socioeconomic status, exposure to ionizing radiation, and other factors have been correlated with an increased incidence of GBM, while female sex hormones, history of allergies, and frequent use of specific drugs might exert protective effects against this disease. However, none of these explain the pathogenesis of GBM. The most recent WHO classification of CNS tumors classifies neoplasms based on their histopathological and molecular characteristics. Modern laboratory techniques, such as matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry, enable the comprehensive metabolic analysis of the tissue sample. MALDI imaging is able to characterize the spatial distribution of a wide array of biomolecules in a sample, in combination with histological features, without sacrificing the tissue integrity. In this review, we first provide an overview of GBM epidemiology, risk, and protective factors, as well as the recent WHO classification of CNS tumors. We then provide an overview of mass spectrometry workflow, with a focus on MALDI imaging, and recent advances in cancer research. Finally, we conclude the review with studies of GBM that utilized MALDI imaging and offer our perspective on future research. Full article

(This article belongs to the Special Issue Molecular Imaging of Cells and Tissues)

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12 pages, 2022 KiB

Open AccessCase Report

Characteristic of Endometrial Stromal Sarcoma by Algorithm of Potential Biomarkers for Uterine Mesenchymal Tumor

by Takuma Hayashi, Kenji Sano, Nobuo Yaegashi, Kaoru Abiko and Ikuo Konishi

Curr. Issues Mol. Biol. 2023, 45(8), 6190-6201; https://doi.org/10.3390/cimb45080390 - 25 Jul 2023

Cited by 2 | Viewed by 1554

Abstract

The benign tumor uterine leiomyoma (UL) develops from the smooth muscle tissue that constitutes the uterus, whereas malignant tumor uterine sarcoma develops from either the smooth muscle tissue or stroma and is different from UL and endometrial cancer. Uterine sarcoma is broadly classified [...] Read more.

The benign tumor uterine leiomyoma (UL) develops from the smooth muscle tissue that constitutes the uterus, whereas malignant tumor uterine sarcoma develops from either the smooth muscle tissue or stroma and is different from UL and endometrial cancer. Uterine sarcoma is broadly classified into three types: uterine leiomyosarcoma, endometrial stromal sarcoma (ESS), and carcinosarcoma. Although uterine leiomyosarcoma and ESS are both classified as uterine sarcoma, they significantly differ in terms of their sites of occurrence, symptoms, and treatment methods. Uterine leiomyosarcoma develops from the muscle tissue constituting the wall of the uterus and accounts for approximately 70% of all uterine sarcoma cases. In contrast, ESS develops from the stromal tissue beneath the endometrium and accounts for approximately 25% of all uterine sarcoma cases. ESS is classified as either low grade (LG) or high grade (HG). This case report aimed to highlight the importance of histopathologic examinations based on surgical specimens. Herein, we reported the case of a 45-year-old woman suspected of having submucosal leiomyoma of the uterus based on imaging results. Transvaginal ultrasonography and endometrial biopsy or partial dilation and curettage were performed. Contrast-enhanced magnetic resonance imaging (MRI) revealed a 32-mm mass projecting from the posterior wall of the uterus into the uterine cavity. T2-weighted imaging revealed a low signal within the mass; thus, submucosal UL was suspected. Histopathologic examination of surgical specimens obtained from a patient suspected of having submucosal UL after contrast-enhanced MRI indicated that the patient had ESS. Despite the remarkable advancements in medical imaging technology, the accuracy of contrast-enhanced MRI for detecting uterine mesenchymal tumors is limited. Therefore, histopathologic diagnosis based on surgical specimens should be performed when medical grounds for diagnosing a benign tumor on contrast-enhanced MRI are lacking. Full article

(This article belongs to the Special Issue Molecular Imaging of Cells and Tissues)

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