64Cu-ATSM Predicts Efficacy of Carbon Ion Radiotherapy Associated with Cellular Antioxidant Capacity

Carbon ion radiotherapy is an emerging cancer treatment modality that has a greater therapeutic window than conventional photon radiotherapy. To maximize the efficacy of this extremely scarce medical resource, it is important to identify predictive biomarkers of higher carbon ion relative biological effectiveness (RBE) over photons. We addressed this issue by focusing on cellular antioxidant capacity and investigated 64Cu(II)-diacetyl-bis(N4-methylthiosemicarbazone) (64Cu-ATSM), a potential radioligand that reflects an over-reduced intracellular environment. We found that the carbon ion RBE correlated with 64Cu-ATSM uptake both in vitro and in vivo. High RBE/64Cu-ATSM cells showed greater steady-state levels of antioxidant proteins and increased capacity to scavenge reactive oxygen species in response to X-rays than low RBE/64Cu-ATSM counterparts; this upregulation of antioxidant systems was associated with downregulation of TCA cycle intermediates. Furthermore, inhibition of nuclear factor erythroid 2-related factor 2 (Nrf2) sensitized high RBE/64Cu-ATSM cells to X-rays, thereby reducing RBE values to levels comparable to those in low RBE/64Cu-ATSM cells. These data suggest that the cellular activity of Nrf2-driven antioxidant systems is a possible determinant of carbon ion RBE predictable by 64Cu-ATSM uptake. These new findings highlight the potential clinical utility of 64Cu-ATSM imaging to identify high RBE tumors that will benefit from carbon ion radiotherapy.

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Identification of Redox-Sensitive Transcription Factors as Markers of Malignant Pleural Mesothelioma

Cancers ◽

10.3390/cancers13051138 ◽

2021 ◽

Vol 13 (5) ◽

pp. 1138

Author(s):

Martina Schiavello ◽

Elena Gazzano ◽

Loredana Bergandi ◽

Francesca Silvagno ◽

Roberta Libener ◽

...

Keyword(s):

Oxidative Stress ◽

Transcription Factors ◽

Malignant Pleural Mesothelioma ◽

Antioxidant Defense ◽

Asbestos Exposure ◽

Predictive Biomarkers ◽

Antioxidant Systems ◽

Pleural Mesothelioma ◽

Related Factor ◽

Aggressive Cancer

Although asbestos has been banned in most countries around the world, malignant pleural mesothelioma (MPM) is a current problem. MPM is an aggressive tumor with a poor prognosis, so it is crucial to identify new markers in the preventive field. Asbestos exposure induces oxidative stress and its carcinogenesis has been linked to a strong oxidative damage, event counteracted by antioxidant systems at the pulmonary level. The present study has been focused on some redox-sensitive transcription factors that regulate cellular antioxidant defense and are overexpressed in many tumors, such as Nrf2 (Nuclear factor erythroid 2-related factor 2), Ref-1 (Redox effector factor 1), and FOXM1 (Forkhead box protein M1). The research was performed in human mesothelial and MPM cells. Our results have clearly demonstrated an overexpression of Nrf2, Ref-1, and FOXM1 in mesothelioma towards mesothelium, and a consequent activation of downstream genes controlled by these factors, which in turn regulates antioxidant defense. This event is mediated by oxidative free radicals produced when mesothelial cells are exposed to asbestos fibers. We observed an increased expression of Nrf2, Ref-1, and FOXM1 towards untreated cells, confirming asbestos as the mediator of oxidative stress evoked at the mesothelium level. These factors can therefore be considered predictive biomarkers of MPM and potential pharmacological targets in the treatment of this aggressive cancer.

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275.Calmodulin-dependent nuclear import pathway of the testis-determining factor SRY

Reproduction Fertility and Development ◽

10.1071/srb04abs275 ◽

2004 ◽

Vol 16 (9) ◽

pp. 275

Author(s):

G. Kaur ◽

A. Delluc-Clavieres ◽

I. Poon ◽

D. A. Jans

Keyword(s):

Nuclear Import ◽

Protein Import ◽

Fluorescent Protein ◽

Fluorescent Proteins ◽

Gonadal Development ◽

Nuclear Accumulation ◽

Related Factor ◽

Testis Development ◽

New Findings

Modulation of the nuclear entry of transcription factors (TFs) and chromatin components is a means by which eukaryotic cells can regulate gene expression in response to extracellular signals and the cell cycle during differentiation and development. TFs and chromatin components access the nucleus through nuclear localisation sequences (NLSs), which mediate interaction with components of the cellular nuclear import machinery, such as members of the importin superfamily. The Ca2+-binding protein calmodulin (CaM ) has previously been shown to bind at or near NLSs in several nuclear-localising proteins that have important roles in testis development including the Y chromosome-encoded HMG-domain-carrying chromatin remodelling factor SRY, and related factor SOX9, both of which are key regulators of gonadal development. SRY function in the nucleus of somatic cells of the fetal gonad, in particular, is essential for development of a testis in males. Here we present new findings implicating a role for CaM in modulating SRY nuclear accumulation, whereby treatment of transfected cells with CaM antagonists significantly reduces nuclear accumulation of green fluorescent protein (GFP)-fusion proteins encoding either full length SRY or the SRY HMG domain alone. An in vitro nuclear transport assay using bacterially expressed fluorescent proteins showed similar results, with native gel electrophoresis/fluorimaging and fluorescence polarisation assays, indicating direct binding of CaM to the SRY HMG domain in Ca2+-dependent fashion. Since clinical mutations resulting in sex reversal occur within SRY's CaM-binding NLS, these results may shed new insight into CaM-dependent pathways of nuclear protein import, and how this may relate to testis development.

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In vitro evaluation of photon and raster-scanned carbon ion radiotherapy in combination with gemcitabine in pancreatic cancer cell lines

Journal of Radiation Research ◽

10.1093/jrr/rrt052 ◽

2013 ◽

Vol 54 (suppl 1) ◽

pp. i113-i119 ◽

Cited By ~ 22

Author(s):

R. A. El Shafie ◽

D. Habermehl ◽

S. Rieken ◽

A. Mairani ◽

L. Orschiedt ◽

...

Keyword(s):

Pancreatic Cancer ◽

Cell Lines ◽

Cancer Cell ◽

Pancreatic Cancer Cell ◽

Cancer Cell Lines ◽

Carbon Ion Radiotherapy ◽

In Vitro Evaluation ◽

Carbon Ion

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Carnosine Protects Macrophages against the Toxicity of Aβ1-42 Oligomers by Decreasing Oxidative Stress

Biomedicines ◽

10.3390/biomedicines9050477 ◽

2021 ◽

Vol 9 (5) ◽

pp. 477

Author(s):

Giuseppe Caruso ◽

Cristina Benatti ◽

Nicolò Musso ◽

Claudia G. Fresta ◽

Annamaria Fidilio ◽

...

Keyword(s):

Oxidative Stress ◽

Therapeutic Potential ◽

Radical Scavenging ◽

Antioxidant Systems ◽

Trypan Blue Exclusion ◽

New Findings ◽

Pre Treatment ◽

The Brain

Carnosine (β-alanyl-L-histidine) is a naturally occurring endogenous peptide widely distributed in excitable tissues such as the brain. This dipeptide has well-known antioxidant, anti-inflammatory, and anti-aggregation activities, and it may be useful for treatment of neurodegenerative disorders such as Alzheimer’s disease (AD). In this disease, peripheral infiltrating macrophages play a substantial role in the clearance of amyloid beta (Aβ) peptides from the brain. Correspondingly, in patients suffering from AD, defects in the capacity of peripheral macrophages to engulf Aβ have been reported. The effects of carnosine on macrophages and oxidative stress associated with AD are consequently of substantial interest for drug discovery in this field. In the present work, a model of stress induced by Aβ1-42 oligomers was investigated using a combination of methods including trypan blue exclusion, microchip electrophoresis with laser-induced fluorescence, flow cytometry, fluorescence microscopy, and high-throughput quantitative real-time PCR. These assays were used to assess the ability of carnosine to protect macrophage cells, modulate oxidative stress, and profile the expression of genes related to inflammation and pro- and antioxidant systems. We found that pre-treatment of RAW 264.7 macrophages with carnosine counteracted cell death and apoptosis induced by Aβ1-42 oligomers by decreasing oxidative stress as measured by levels of intracellular nitric oxide (NO)/reactive oxygen species (ROS) and production of peroxynitrite. This protective activity of carnosine was not mediated by modulation of the canonical inflammatory pathway but instead can be explained by the well-known antioxidant and free-radical scavenging activities of carnosine, enhanced macrophage phagocytic activity, and the rescue of fractalkine receptor CX3CR1. These new findings obtained with macrophages challenged with Aβ1-42 oligomers, along with the well-known multimodal mechanism of action of carnosine in vitro and in vivo, substantiate the therapeutic potential of this dipeptide in the context of AD pathology.

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Effect of Cooking Methods on the Antioxidant Capacity of Plant Foods Submitted to In Vitro Digestion–Fermentation

Antioxidants ◽

10.3390/antiox9121312 ◽

2020 ◽

Vol 9 (12) ◽

pp. 1312

Author(s):

Beatriz Navajas-Porras ◽

Sergio Pérez-Burillo ◽

Álvaro Jesús Valverde-Moya ◽

Daniel Hinojosa-Nogueira ◽

Silvia Pastoriza ◽

...

Keyword(s):

Antioxidant Capacity ◽

Thermal Processing ◽

In Vitro Digestion ◽

Antioxidant Systems ◽

Cooking Methods ◽

Antioxidant Compounds ◽

Plant Foods ◽

Total Antioxidant ◽

Endogenous Antioxidant

The antioxidant capacity of foods is essential to complement the body’s own endogenous antioxidant systems. The main antioxidant foods in the regular diet are those of plant origin. Although every kind of food has a different antioxidant capacity, thermal processing or cooking methods also play a role. In this work, the antioxidant capacity of 42 foods of vegetable origin was evaluated after in vitro digestion and fermentation. All foods were studied both raw and after different thermal processing methods, such as boiling, grilling roasting, frying, toasting and brewing. The cooking methods had an impact on the antioxidant capacity of the digested and fermented fractions, allowing the release and transformation of antioxidant compounds. In general, the fermented fraction accounted for up to 80–98% of the total antioxidant capacity. The most antioxidant foods were cocoa and legumes, which contributed to 20% of the daily antioxidant capacity intake. Finally, it was found that the antioxidant capacity of the studied foods was much higher than those reported by other authors since digestion–fermentation pretreatment allows for a higher extraction of antioxidant compounds and their transformation by the gut microbiota.

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Impact of Hypoxia on Carbon Ion Therapy in Glioblastoma Cells: Modulation by LET and Hypoxia-Dependent Genes

Cancers ◽

10.3390/cancers12082019 ◽

2020 ◽

Vol 12 (8) ◽

pp. 2019 ◽

Cited By ~ 2

Author(s):

Samuel Valable ◽

Aurélie N. Gérault ◽

Gaëlle Lambert ◽

Marine M. Leblond ◽

Clément Anfray ◽

...

Keyword(s):

Tumor Hypoxia ◽

Physical Factors ◽

X Rays ◽

Erk Activation ◽

Carbon Ion ◽

Hypoxic Environment ◽

Ion Therapy ◽

High Let ◽

A Cell

Tumor hypoxia is known to limit the efficacy of ionizing radiations, a concept called oxygen enhancement ratio (OER). OER depends on physical factors such as pO2 and linear energy transfer (LET). Biological pathways, such as the hypoxia-inducible transcription factors (HIF), might also modulate the influence of LET on OER. Glioblastoma (GB) is resistant to low-LET radiation (X-rays), due in part to the hypoxic environment in this brain tumor. Here, we aim to evaluate in vitro whether high-LET particles, especially carbon ion radiotherapy (CIRT), can overcome the contribution of hypoxia to radioresistance, and whether HIF-dependent genes, such as erythropoietin (EPO), influence GB sensitivity to CIRT. Hypoxia-induced radioresistance was studied in two human GB cells (U251, GL15) exposed to X-rays or to carbon ion beams with various LET (28, 50, 100 keV/µm), and in genetically-modified GB cells with downregulated EPO signaling. Cell survival, radiobiological parameters, cell cycle, and ERK activation were assessed under those conditions. The results demonstrate that, although CIRT is more efficient than X-rays in GB cells, hypoxia can limit CIRT efficacy in a cell-type manner that may involve differences in ERK activation. Using high-LET carbon beams, or targeting hypoxia-dependent genes such as EPO might reduce the effects of hypoxia.

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Protective Effect of Cocoa Bean Shell against Intestinal Damage: An Example of Byproduct Valorization

Antioxidants ◽

10.3390/antiox10020280 ◽

2021 ◽

Vol 10 (2) ◽

pp. 280

Author(s):

Daniela Rossin ◽

Letricia Barbosa-Pereira ◽

Noemi Iaia ◽

Barbara Sottero ◽

Alice Costanza Danzero ◽

...

Keyword(s):

Antioxidant Capacity ◽

Inflammatory Responses ◽

Radical Scavenging ◽

Ice Cream ◽

Cholesterol Oxidation ◽

Cocoa Bean ◽

Related Factor ◽

Intestinal Damage ◽

Food Ingredient

Background: Cocoa bean shell (CBS), a main byproduct of cocoa processing, represents a source of components such as polyphenols and methylxanthines, which have been associated with a reduced risk of several diseases. Therefore, CBS has potential application as a food ingredient. Intestinal mucosa is exposed to immune and inflammatory responses triggered by dietary agents, such as oxysterols, which derive from cholesterol oxidation and are pro-oxidant compounds able to affect intestinal function. We aimed at investigating the capability of the Forastero cultivar CBS, added or not added to ice cream, to protect against the intestinal barrier damage induced by a dietary oxysterol mixture. Methods: Composition and antioxidant capacity of in vitro digested CBS and CBS-enriched ice cream were analyzed by high-performance liquid chromatography and 1,1-diphenyl-2-picryl-hydrazyl radical-scavenging assay, respectively. CaCo-2 cells differentiated into enterocyte-like monolayer were incubated with 60 µM oxysterol mixture in the presence of CBS formulations. Results: The oxysterol mixture induced tight junction impairment, interleukin-8 and monocyte chemoattractant protein-1 cell release, and oxidative stress-related nuclear factor erythroid 2 p45-related factor 2 response Nrf2. Both CBSs protected cells from these adverse effects, probably thanks to their high phenolic content. CBS-enriched ice cream showed the highest antioxidant capacity. Theobromine, which is in high concentrations of CBS, was also tested. Although theobromine exerted no effect on Nrf2 expression, its anti-inflammatory cooperating activity in CBS effect cannot be excluded. Conclusions: Our findings suggest that CBS-enriched ice cream may be effective in the prevention of gut integrity damage associated with oxidative/inflammatory reactions.

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Effect of Cooking Methods on the Antioxidant Capacity of Foods of Animal Origin Submitted to In Vitro Digestion-Fermentation

Antioxidants ◽

10.3390/antiox10030445 ◽

2021 ◽

Vol 10 (3) ◽

pp. 445

Author(s):

Beatriz Navajas-Porras ◽

Sergio Pérez-Burillo ◽

Álvaro Valverde-Moya ◽

Daniel Hinojosa-Nogueira ◽

Silvia Pastoriza ◽

...

Keyword(s):

Antioxidant Capacity ◽

In Vitro Digestion ◽

Bioactive Molecules ◽

Animal Origin ◽

Antioxidant Systems ◽

Cooking Methods ◽

Antioxidant Compounds ◽

Microbial Fermentation ◽

Endogenous Antioxidant

The human body is exposed to oxidative damage to cells and though it has some endogenous antioxidant systems, we still need to take antioxidants from our diet. The main dietary source of antioxidants is vegetables due to their content of different bioactive molecules. However, there are usually other components of the diet, such as foods of animal origin, that are not often linked to antioxidant capacity. Still, these foods are bound to exert some antioxidant capacity thanks to molecules released during gastrointestinal digestion and gut microbial fermentation. In this work, the antioxidant capacity of 11 foods of animal origin has been studied, submitted to different culinary techniques and to an in vitro digestion and gut microbial fermentation. Results have shown how dairy products potentially provide the highest antioxidant capacity, contributing to 60% of the daily antioxidant capacity intake. On the other hand, most of the antioxidant capacity was released during gut microbial fermentation (90–98% of the total antioxidant capacity). Finally, it was found that the antioxidant capacity of the studied foods was much higher than that reported by other authors. A possible explanation is that digestion–fermentation pretreatment allows for a higher extraction of antioxidant compounds and their transformation by the gut microbiota. Therefore, although foods of animal origin cannot be compared to vegetables in the concentration of antioxidant molecules, the processes of digestion and fermentation can provide some, giving animal origin food some qualities that could have been previously unappreciated.

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Sodium Tanshinone IIA Sulfonate Attenuates Erectile Dysfunction in Rats with Hyperlipidemia

Oxidative Medicine and Cellular Longevity ◽

10.1155/2020/7286958 ◽

2020 ◽

Vol 2020 ◽

pp. 1-13 ◽

Cited By ~ 1

Author(s):

Liren Zhong ◽

Wei Ding ◽

Qingyu Zeng ◽

Binglin He ◽

Haibo Zhang ◽

...

Keyword(s):

Oxidative Stress ◽

Erectile Dysfunction ◽

Antioxidant Capacity ◽

Erectile Function ◽

Tanshinone Iia ◽

Heme Oxygenase 1 ◽

Related Factor ◽

Sprague Dawley ◽

Sodium Tanshinone Iia Sulfonate

Hyperlipidemia is considered one of the most important risk factors for erectile dysfunction (ED). To determine the effect of sodium tanshinone IIA sulfonate (STS) as an antioxidant agent on ED in high-fat diet- (HFD-) induced hyperlipidemia in rats and to investigate if STS administration could improve erectile function via hydrogen sulfide (H2S) production by inhibition of oxidative stress. Hyperlipidemia was induced in Sprague-Dawley rats by feeding HFD for 16 weeks. The rats were randomly divided into 3 groups: control, HFD, and HFD treated with STS (10 mg/kg/day for 12 weeks, intraperitoneal injection). Erectile function including intracavernosal pressure (ICP), H2S production, and antioxidant capacity was assessed. In addition, cavernosal smooth muscle cells (CSMC) isolated from SD rats were pretreated with STS in vitro and exposed to H2O2. Expressions of nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1), activity of antioxidant enzymes, and H2S-generating enzymes within CSMC were examined. ICP was significantly decreased in HFD rats compared with control. In addition, decreased H2S production and expression of cystathionine ɣ-lyase (CSE) and cystathionine β-synthase (CBS) associated with increased oxidative stress were observed in the penile tissue of HFD rats. However, all these changes were reversed by 16 weeks after STS administration. STS also increased antioxidant defense as evidenced by increased expression of Nrf2/HO-1 in the penile tissue of HFD rats. In CSMC, pretreatment with STS attenuated the decreased expression of CSE and CBS and H2S production by H2O2. STS exerted similar protective antioxidative effect as shown in the in vivo hyperlipidemia model. The present study demonstrated the redox effect of STS treatment on ED via increased H2S production in HFD-induced hyperlipidemia rat model by increased antioxidant capacity via activation of the Nrf2/HO-1 pathway, which provides STS potential clinical application in the treatment of hyperlipidemia-related ED.

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