miR-199a Downregulation as a Driver of the NOX4/HIF-1α/VEGF-A Pathway in Thyroid and Orbital Adipose Tissues from Graves′ Patients

Graves’ disease (GD) is an autoimmune thyroiditis often associated with Graves’ orbitopathy (GO). GD thyroid and GO orbital fat share high oxidative stress (OS) and hypervascularization. We investigated the metabolic pathways leading to OS and angiogenesis, aiming to further decipher the link between local and systemic GD manifestations. Plasma and thyroid samples were obtained from patients operated on for multinodular goiters (controls) or GD. Orbital fats were from GO or control patients. The NADPH-oxidase-4 (NOX4)/HIF-1α/VEGF-A signaling pathway was investigated by Western blotting and immunostaining. miR-199a family expression was evaluated following quantitative real-time PCR and/or in situ hybridization. In GD thyroids and GO orbital fats, NOX4 was upregulated and correlated with HIF-1α stabilization and VEGF-A overexpression. The biotin assay identified NOX4, HIF-1α and VEGF-A as direct targets of miR-199a-5p in cultured thyrocytes. Interestingly, GD thyroids, GD plasmas and GO orbital fats showed a downregulation of miR-199a-3p/-5p. Our results also highlighted an activation of STAT-3 signaling in GD thyroids and GO orbital fats, a transcription factor known to negatively regulate miR-199a expression. We identified NOX4/HIF-1α/VEGF-A as critical actors in GD and GO. STAT-3-dependent regulation of miR-199a is proposed as a common driver leading to these events in GD thyroids and GO orbital fats.

THE EFFECTS OF OXIDATIVE STRESS ON THE EYES OF SPRAGUE–DAWLEY RATS

An imbalance between oxidants and antioxidants in favor of the oxidants, potentially leading to damage, is termed as ‘oxidative stress’. Some studies indicate that reactive oxygen species (ROS) may increase in resistance of the aqueous humor drainage or induce the rearrangement of human trabecular meshwork and result in glaucoma finally. An eight-week d-galactose injection was used to mimic high oxidative stress rats. Behavior tests, fundus images, anterior chamber depth, anterior chamber diameter (ACDia) and pupil diameter (PD) were investigated. Intraocular pressure (IOP) measurement and serum were taken for antioxidant capability of superoxide dismutase (SOD) and the amount of nitric oxide (NO). After perfusion, Müller’s cells and astrocyte, microglial and inducible Nitric Oxide Synthase (iNOS) were labeled. The activities of SOD were decreased after induction. The spatial memory ability and muscular endurance were impaired through Morris water maze (MWM) and weight-loaded forced swimming test. These results demonstrated that oxidative stress was exacerbated by d-galactose. The IOP was gradually increased in the third week after d-galactose induction. Immunohistochemical stains showed that the density of Müller’s cells, astrocytes and microglia increased in d-galactose group. Microglia mostly became active. These data indicate that oxidative stress may result in the impairment of behavior, increase of Müller’s cells, astrocyte and microglia which is induced by d-galactose. The correlation between oxidative stress and elevation of IOP should be further studied. These may involve in the pathologies of aging or other neurodegenerative disease.

A Review on Lactoferrin and Central Nervous System Diseases

Central nervous system (CNS) diseases are currently one of the major health issues around the world. Most CNS disorders are characterized by high oxidative stress levels and intense inflammatory responses in affected tissues. Lactoferrin (Lf), a multifunctional iron-binding glycoprotein, plays a significant role in anti-inflammatory, antibacterial, antiviral, reactive oxygen species (ROS) modulator, antitumor immunity, and anti-apoptotic processes. Previous studies have shown that Lf is abnormally expressed in a variety of neurological diseases, especially neurodegenerative diseases. Recently, the promotion of neurodevelopment and neuroprotection by Lf has attracted widespread attention, and Lf could be exploited both as an active therapeutic agent and drug nanocarrier. However, our understanding of the roles of Lf proteins in the initiation or progression of CNS diseases is limited, especially the roles of Lf in regulating neurogenesis. This review highlights recent advances in the understanding of the major pharmacological effects of Lf in CNS diseases, including neurodegenerative diseases, cerebrovascular disease, developmental delays in children, and brain tumors.

A new classification of cardio-oncology syndromes

Increasing evidence suggests a multifaceted relationship exists between cancer and cardiovascular disease (CVD). Here, we introduce a 5-tier classification system to categorize cardio-oncology syndromes (COS) that represent the aspects of the relationship between cancer and CVD. COS Type I is characterized by mechanisms whereby the abrupt onset or progression of cancer can lead to cardiovascular dysfunction. COS Type II includes the mechanisms by which cancer therapies can result in acute or chronic CVD. COS Type III is characterized by the pro-oncogenic environment created by the release of cardiokines and high oxidative stress in patients with cardiovascular dysfunction. COS Type IV is comprised of CVD therapies and diagnostic procedures which have been associated with promoting or unmasking cancer. COS Type V is characterized by factors causing systemic and genetic predisposition to both CVD and cancer. The development of this framework may allow for an increased facilitation of cancer care while optimizing cardiovascular health through focused treatment targeting the COS type.

Effects of Binahong (Anredera cordifolia (Tenore) Steenis) Extracts on the Levels of Malondialdehyde (MDA) in Cataract Goat Lenses

Cataracts are one of the most causes of blindness in the world. Oxidative stress can form pathological conditions such as cataracts. This oxidative stress ability can be measured by the malondialdehyde (MDA) biomarker. Binahong leaves (Anredera cordifolia (Tenore) Steenis) are native plants from Indonesia that are used to treat various diseases including cataract treatment. Binahong leaf (Anredera cordifolia (Tenore) Steenis) has a high amount of flavonoids and is rich in antioxidants that can be used to treat cataracts. Objective. The purpose of this study was to assess the effect of binahong leaf extract on the levels of MDA in a goat lens with cataract-induced material. Method. As many as possible, 40 goat eye lenses were divided into several groups, namely, group I normal lenses as controls (glucose 5.5 mM), group II lenses were cataract induced with glucose concentration of 55 mM, group III lenses with glucose 55 mM + binahong leaf extract (100 μg/ml), group IV lens with glucose 55 mM + binahong leaf extract (200 μg/ml), and group V lens with glucose 55 mM + quercetin (positive control). Biochemical parameters measured in the lens homogenate are malondialdehyde lens morphology in all groups’ observations and comparisons made. Results. The results of the study found that the lens group with the addition of binahong extract showed more results transparency compared to lens groups induced by glucose concentrations of 55 mM). This shows that the diabetic cataract group experienced high oxidative stress due to the accumulation of sorbitol compounds derived from glucose which caused turbidity in the goat eye lens and increased levels of lens MDA. Binahong levels at concentrations of 100 or 200 can inhibit MDA production. Conclusion. Binahong (Anredera cordifolia (Tenore) Steenis) extract has the ability to inhibit the production of MDA levels. In glucose-induced goat lenses, binahong extract and quercetin show antioxidant and anticataract properties.

Protective Effects of Fucoxanthin on Hydrogen Peroxide-Induced Calcification of Heart Valve Interstitial Cells

Cardiovascular diseases such as atherosclerosis and aortic valve sclerosis involve inflammatory reactions triggered by various stimuli, causing increased oxidative stress. This increased oxidative stress causes damage to the heart cells, with subsequent cell apoptosis or calcification. Currently, heart valve damage or heart valve diseases are treated by drugs or surgery. Natural antioxidant products are being investigated in related research, such as fucoxanthin (Fx), which is a marine carotenoid extracted from seaweed, with strong antioxidant, anti-inflammatory, and anti-tumor properties. This study aimed to explore the protective effect of Fx on heart valves under high oxidative stress, as well as the underlying mechanism of action. Rat heart valve interstitial cells under H2O2-induced oxidative stress were treated with Fx. Fx improved cell survival and reduced oxidative stress-induced DNA damage, which was assessed by cell viability analysis and staining with propidium iodide. Alizarin Red-S analysis indicated that Fx has a protective effect against calcification. Furthermore, Western blotting revealed that Fx abrogates oxidative stress-induced apoptosis via reducing the expression of apoptosis-related proteins as well as modulate Akt/ERK-related protein expression. Notably, in vivo experiments using 26 dogs treated with 60 mg/kg of Fx in combination with medical treatment for 0.5 to 2 years showed significant recovery in their echocardiographic parameters. Collectively, these in vitro and in vivo results highlight the potential of Fx to protect heart valve cells from high oxidative stress-induced damage.

Apple Pomace as a Source of Bioactive Polyphenol Compounds in Gluten-Free Breads

Gluten-free products based on starch and hydrocolloids are deficient in nutrients and do not contain pro-health substances. Therefore, they should be enriched in raw materials naturally rich in antioxidants, especially if they are intended for celiac patients, prone to high oxidative stress. Apart from the traditionally used pseudo-cereals, seeds, vegetables and fruits, innovative substrates such as the by-product (especially in Poland) dry apple pomace could be applied. The study material consisted of gluten-free bread enriched with apple pomace. The content of individual polyphenols, the content of total polyphenol and flavonoids, and also the antioxidant potential of the bread were determined by the UPLC-PDA-MS/MS methods. It was observed that apple pomace was a natural concentrate of bioactive substances from the group of polyphenols. In summary, gluten-free bread with 5% content of apple pomace showed the highest organoleptic scores and contained high levels of phenolic compounds. The values of total phenolic content, and the amounts of flavonoids, phenolic acids and phloridzin in this bread were 2.5, 8, 4 and 21 times higher in comparison to control.

Multifunctional Molecule, JM-20, Reverses Aluminum Chloride-Induced Memory Impairment and Neuronal Damage in Rats

Alzheimer's disease (AD) is a neurodegenerative disease that worsens with aging. Today, there is a worldwide effort to find new drugs that could delay the onset, slow the progression, or improve symptoms of AD. Oral administration of aluminum to rodents recapitulates some pathological alterations observed in AD, being considered a convenient tool for modeling and testing the efficacy of new therapeutics. Our previous studies have shown that JM-20, a dihydropyridine and benzodiazepine hybrid molecule protected memory in an environment with cholinergic dysfunction, high oxidative stress, hyperactivation of acetylcholinesterase (AChE) enzyme, and mitochondrial damage produced by scopolamine. In order to gain further insight into the effects on JM-20 on AD pathology, we evaluated the protective effects of JM-20 after chronic AlCl3 administration to rats, and assessed several types of episodic memory alterations and associated-pathological mechanisms, including mitochondrial dysfunction, AChE hyperactivity, inflammation, and apoptosis-related proteins. We used behavioral tasks to test spatial, working an emotional- associative memory, as well as molecular, enzymatic and histological assays to evaluate selected biochemical parameters. Our study showed that JM-20 prevented memory decline alongside with the inhibition of aluminum-induced alterations of oxidative parameters, and increase of AChE activity and of tumor necrosis factor alpha (TNF-α) levels. JM-20 also preserved anti-apoptotic proteins and protected against axonal and neuronal damaged in the hippocampus and prefrontal cortex. Altogether, our findings expanded our understanding of the ability of JM-20 to preserve essential types of memory in rats under neurotoxic conditions, and suggest its potential capacity to counteract etiological factors of AD by breaking the progression of key neurodegeneration-associated steps in a rat model of the disease.

Iron acquisition across outer membrane: The role of major outer membrane protein Slr1908 under iron supplemented conditions in Synechocystis 6803

The Ton-B dependent outer membrane (OM) transporters are responsible for active iron (Fe) import in Synechocystis sp. strain PCC 6803 (S. 6803 or WT) under Fe depletion. However, the mechanism of Fe acquisition under Fe supplemented conditions remains uncharacterised. In the present study, functional role of OMP Slr1908 in S. 6803 was addressed by insertional mutagenesis. The Δslr1908 cells exhibited slower growth in the first week in comparison to the WT and displayed an absorption and 77K fluorescence spectrum typical of Fe deficiency. Indeed, the mutant had ∼ 80% less Fe as confirmed by atomic absorption spectroscopy and 55Fe-radiotracer uptake. The iron deficiency was paralleled with low Mn content. The mutant had low SOD content as well as activity, less cytochromes, less chlorophyll content, less Fv/ Fm, lower ETRII and high oxidative stress in comparison to the WT at the end of first week. Interestingly, the mutant showed transcriptional upregulation of iron stress induced protein isiA and isiB signifying intracellular Fe deficiency. Upregulation of OMP Slr0042 was also observed at RNA and protein level. The results indicate that Slr1908 is a major Fe uptake OMP in S. 6803 the deletion of which leads to initial slow growth that gets partially offset by induction of other Fe importing OMPs.