Impact of ozone therapy on mouse liver mitochondrial function and antioxidant system.

Ozone therapy's efficacy might stem from the regulated and mild oxidative stress resulting from ozone's interactions with various biological elements. The present work aimed to characterize the hepatic mitochondrial response to ozone treatment and its relationship with the antioxidant system response. Two groups of mice were used: one control group and another injected intraperitoneally with an O3/O2 mixture (80 ml/kg) for 5 days. Mitochondrial respiration supported by different substrates was significantly inhibited, as well as complexes I and II/III, but not complex IV. The analysis of the electron transport chain complex activity showed significant inhibitions in complexes I and II/III but not in complex IV. These inhibitions can prevent mitochondrial reactive oxygen species (ROS) production. Additionally, there was a decline in glutathione content, unaccompanied by a rise in its oxidized form. The ozone-treated groups showed a significant increase in the activity of superoxide dismutase and glutathione peroxidase, while catalase and glutathione reductase experienced no significant alterations. Adenine nucleotides increased in the ozone group, but only the increase in adenosine diphosphate is significant, so the cell's energy charge is unaffected. This study shows that mitochondria may play a crucial role in ozone treatment. However, it also highlights the need for further studies to understand the molecular mechanism.

Mitochondrial Dysfunction in Skeletal Muscle of Rotenone-Induced Rat Model of Parkinson's Disease: SC-Nanophytosomes as Therapeutic Approach.

The development of new therapeutic options for Parkinson's disease (PD) requires formulations able to mitigate both brain degeneration and motor dysfunctions. SC-Nanophytosomes, an oral mitochondria-targeted formulation developed with Codium tomentosum membrane polar lipids and elderberry anthocyanin-enriched extract, promote significant brain benefits on a rotenone-induced rat model of PD. In the present work, the effects of SC-Nanophytosome treatment on the skeletal muscle tissues are disclosed. It is unveiled that the rotenone-induced PD rat model exhibits motor disabilities and skeletal muscle tissues with deficient activity of mitochondrial complexes I and II along with small changes in antioxidant enzyme activity and skeletal muscle lipidome. SC-Nanophytosome treatment mitigates the impairment of complexes I and II activity, improving the mitochondrial respiratory chain performance at levels that surpass the control. Therefore, SC-Nanophytosome competence to overcome the PD-related motor disabilities should be also associated with its positive outcomes on skeletal muscle mitochondria. Providing a cellular environment with more reduced redox potential, SC-Nanophytosome treatment improves the skeletal muscle tissue's ability to deal with oxidative stress stimuli. The PD-related small changes on skeletal muscle lipidome were also counteracted by SC-Nanophytosome treatment. Thus, the present results reinforces the concept of SC-Nanophytosomes as a mitochondria-targeted therapy to address the neurodegeneration challenge.

Assessing micro and nanoplastics toxicity using rodent models: Investigating potential mitochondrial implications.

Mitochondria's role as a central hub in cellular metabolism and signaling cascades is well established in the scientific community, being a classic marker of organisms' response to toxicant exposure. Nonetheless, little is known concerning the effects of emerging contaminants, such as microplastics, on mitochondrial metabolism. Micro- and nanoplastics present one of the major problems faced by modern societies. What was once an environmental problem is now recognized as an one-health issue, but little is known concerning microplastic impact on human health. Indeed, only recently, human exposure to microplastics was acknowledged by the World Health Organization, resulting in a growing interest in this research topic. Nonetheless, the mechanisms behind micro- and nanoplastics toxicity are yet to be understood. Animal models, nowadays, are the most appropriate approach to uncovering this knowledge gap. In the present review article, we explore investigations from the last two years using rodent models and reach to find the molecular mechanism behind micro- and nanoplastics toxicity and if mitochondria can act as a target. Although no research article has addressed the effects of mitochondria yet, reports have highlighted molecular and biochemical alterations that could be linked to mitochondrial function. Furthermore, certain studies described the effects of disruptions in mitochondrial metabolism, such as oxidative stress. Micro- and nanoplastics may, directly and indirectly, affect this vital organelle. Investigations concerning this topic should be encouraged once they can bring us closer to understanding the mechanisms underlying these particles' harmful effects on human health.

The administration of methyl and butyl parabens interferes with the enzymatic antioxidant system and induces genotoxicity in rat testis: possible relation to male infertility.

Parabens are esters of p-hydroxybenzoic acid, used for decades as a preservative in many products, including agrochemicals, pharmaceuticals, foods and cosmetics. Concerns regarding parabens toxicity include adverse effects on endocrine activity, carcinogenesis, infertility, spermatogenesis, and adipogenesis. The present study aimed to investigate the in vivo administration of methyl and butylparaben at concentrations of 100 and 200 mg/kg body weight, by subcutaneous injection, in variable murinometric measurements, antioxidant systems and genotoxicity. The administration of parabens did not affect the consumption of water and food. However, there was a decrease in the weight of the testes and the seminal vesicle (p < 0.05). The administration of parabens caused an increase in superoxide dismutase for methylparaben (200 mg/kg) and both concentrations of butylparaben (p < 0.05). Catalase showed increased activity in all groups treated with parabens. In contrast, glutathione reductase and glutathione S-transferase suffered a decrease in the groups treated with both parabens. These results show that parabens, especially butyl, can affect the rat testis enzymatic antioxidant system, decreasing the cellular antioxidant capacity, which was confirmed by the decrease in the glutathione reducing power, expressed by the reduced glutathione/oxidized glutathione ratio. Therefore, an increase in lipid peroxidation was observed, which was significant in the case of butyl. Genetic Damage Indicator values show that butylparaben treatments displayed significantly higher values than the control. This study shows for the first time that parabens can induce genotoxicity in the rat male reproductive organ.

Effects of chronic exposure of naturally weathered microplastics on oxidative stress level, behaviour, and mitochondrial function of adult zebrafish (Danio rerio).

Microplastics (MPs) are a big and growing environmental concern, with studies showing sublethal to acute biological impacts on typical aquatic organisms. However, little is known about the biological effects of naturally weathered MPs, particularly focusing on mitochondria dysfunction as the key trigger of the biological effects. Therefore, in this study, naturally weathered MPs were produced from day-to-day life products, characterized, and chronically exposed (21 days) to adult zebrafish at the concentration of 0.1 and 1 mg/L. Locomotion and unconditioned anxiety-like behaviour was assessed. Mitochondrial respiration, membrane potential, mitochondrial complex activity and oxidative-related parameters were evaluated in the brain and liver. The results revealed the weathered MPs as a copolymer of propylene and ethylene that induced anxiety-like behaviour. There was an increase in brain catalase activity while the brain lactate dehydrogenase activity was inhibited after exposure to 1 mg/L. Brain glutathione levels were increased while their ratio was not affected. Mitochondrial respiratory chain complex Ⅱ and IV were also significantly decreased in the brain, although not compromising mitochondrial function. On the other hand, exposure to 1 mg/L caused a deficiency in liver mitochondrial respiration and decreased mitochondrial membrane potential, which were associated with the mitochondrial respiratory chain inhibition. An increase in hepatic superoxide dismutase and catalase activity was noticed, supporting the occurrence of ROS-induced ROS release as the potential trigger for the mitochondrial dysfunction. Overall, these findings highlight the potential indirect and cumulative environmental effects these particles may pose to aquatic ecosystems.

Long-Term Aerobic Training Improves Mitochondrial and Antioxidant Function in the Liver of Wistar Rats Preventing Hepatic Age-Related Function Decline.

Most studies on the effects of physical exercise have focused on its influence on muscle tissue, forgetting its interference in liver function. Ageing leads to the progressive impairment of hepatic functions. Several biochemical and bioenergetics parameters were determined to test the impact of a lifelong aerobic training program in the hepatic age-related and the development of an adaptative response. Liver samples were collected from 28 male Wistar rats (4-week-old, 159.4 ± 11.9 g at the beginning of the protocol), randomly distributed into two groups: non-exercised or exercised and submitted to a treadmill exercise program (60 min/day, 5 days/week, at 70% of maximal running speed), for 24 (n = 9) or 54 weeks (n = 10). A maximal running speed test was performed to determine the training speed. Antioxidant enzyme activity, cellular redox status, oxidative stress, mitochondrial respiratory chain enzymes and respiratory activity were performed in liver samples. Lifelong exercise decreased the age-associated decline in mitochondrial dysfunction, increasing the respiratory rate in state 2 (mitochondrial respiration stimulated by the substrate in the absence of added ADP) (p = 0.03) and citrate synthase enzymatic activity (p = 0.007). Complex II (p < 0.0001) and IV (p < 0.001) showed a decrease in enzymatic activity. Ageing-related oxidative stress was also attenuated by physical exercise, as showed by the increase in first-line defense antioxidant enzymes (superoxide dismutase (p = 0.07) and catalase (p = 0.03)), decreased lipid peroxidation levels (p = 0.864 for total fraction, p = 0,27 for mitochondrial fraction) and higher glutathione reduced/oxidized ratio (p = 0.02). According to our results, the regular practice of exercise can prevent the liver's mitochondrial dysfunction and loss of antioxidant system efficacy that may arise from ageing, highlighting the benefit of lifelong aerobic exercise in preventing age-related hepatic impairment and associated diseases.

Pulegone and Eugenol Oral Supplementation in Laboratory Animals: Results from Acute and Chronic Studies.

Essential oils are natural compounds used by humans for scientific purposes due to their wide range of properties. Eugenol is mostly present in clove oil, while pulegone is the main constituent of pennyroyal oil. To guarantee the safe use of eugenol and pulegone for both humans and animals, this study addressed, for the first time, the effects of these compounds, at low doses (chronic toxicity) and high doses (acute toxicity), in laboratory animals. Thirty-five FVB/n female mice were randomly assigned to seven groups (n = 5): group I (control, non-additive diet); group II (2.6 mg of eugenol + 2.6 mg of pulegone); group III (5.2 mg of eugenol + 5.2 mg of pulegone); group IV (7.8 mg of eugenol + 7.8 mg of pulegone); group V (7.8 mg of eugenol); group VI (7.8 mg of pulegone); and group VII (1000 mg of eugenol + 1000 mg of pulegone). The compounds were administered in the food. Groups I to VI were integrated into the chronic toxicity study, lasting 28 days, and group VII was used in the acute toxicity study, lasting 7 days. Animals were monitored to assess their general welfare. Water and food intake, as well as body weight, were recorded. On the 29th day, all animals were euthanized by an overdose of ketamine and xylazine, and a complete necropsy was performed. Blood samples were collected directly from the heart for microhematocrit and serum analysis, as well as for comet assay. Organs were collected, weighed, and fixed in formaldehyde for further histological analysis and enzymatic assay. Eugenol and pulegone induced behavioral changes in the animals, namely in the posture, hair appearance and grooming, and in mental status. These compounds also caused a decrease in the animals' body weight, as well as in the food and water consumption. A mortality rate of 20% was registered in the acute toxicity group. Both compounds modulated the serum levels of triglycerides and alanine aminotransferase. Eugenol and pulegone induced genetic damage in all animals. Eugenol increased the activity of the CAT enzyme. Both compounds increased the GR enzyme at the highest dose. Moreover, pulegone administered as a single compound increased the activity of the GST enzyme. Histopathological analysis revealed inflammatory infiltrates in the lungs of groups II, III, and IV. The results suggest that eugenol and pulegone may exert beneficial or harmful effects, depending on the dose, and if applied alone or in combination.

Brain Effects of SC-Nanophytosomes on a Rotenone-Induced Rat Model of Parkinson's Disease-A Proof of Concept for a Mitochondria-Targeted Therapy.

Mitochondria are an attractive target to fight neurodegenerative diseases due to their important functions for cells and the particularly close relationship between the functional connectivity among brain regions and mitochondrial performance. This work presents a mitochondria-targeted therapy designed to modulate the functionality of the mitochondrial respiratory chain and lipidome, parameters that are affected in neurodegeneration, including in Parkinson's disease (PD). This therapy is supported by SC-Nanophytosomes constructed with membrane polar lipids, from Codium tomentosum, and elderberry anthocyanin-enriched extract, from Sambucus nigra L. SC-Nanophytosomes are nanosized vesicles with a high negative surface charge that preserve their properties, including anthocyanins in the flavylium cation form, under conditions that mimic the gastrointestinal tract pH changes. SC-Nanophytosomes, 3 µM in phospholipid, and 2.5 mg/L of EAE-extract, delivered by drinking water to a rotenone-induced PD rat model, showed significant positive outcomes on disabling motor symptoms associated with the disease. Ex vivo assays were performed with two brain portions, one comprising the basal ganglia and cerebellum (BG-Cereb) and the other with the cerebral cortex (C-Cortex) regions. Results showed that rotenone-induced neurodegeneration increases the α-synuclein levels in the BG-Cereb portion and compromises mitochondrial respiratory chain functionality in both brain portions, well-evidenced by a 50% decrease in the respiratory control rate and up to 40% in complex I activity. Rotenone-induced PD phenotype is also associated with changes in superoxide dismutase and catalase activities that are dependent on the brain portion. Treatment with SC-Nanophytosomes reverted the α-synuclein levels and antioxidant enzymes activity to the values detected in control animals. Moreover, it mitigated mitochondrial dysfunction, with positive outcomes on the respiratory control rate, the activity of individual respiratory complexes, and the fatty acid profile of the membrane phospholipids. Therefore, SC-Nanophytosomes are a promising tool to support mitochondria-targeted therapy for neurodegenerative diseases.

Multicomponent exercise program effects on fitness and cognitive function of elderlies with mild cognitive impairment: Involvement of oxidative stress and BDNF.

Regular exercise has been shown to be one of the most important lifestyle influences on improving functional performance, and decreasing morbidity and all-cause mortality among older people. However, although there is some evidence on the effects of aerobic training on oxidative stress, there is little information regarding the effects of multicomponent exercise (dual-task training) and combination of exercise with cognitive stimulation on oxidative stress. In this context, the aim of this study was to verify the effects of a multicomponent exercise program on physical fitness and cognitive function in the elderly with mild cognitive impairment and determine the role of oxidative stress and brain-derived neurotrophic factor (BDNF). At baseline, 37 elderly nursing home residents with mild cognitive impairment were divided into two groups: the control group (CG, n = 12, 81.8 years) and the experimental group (EG, n = 25, 83.2 years). These elderlies followed multicomponent exercise training for 24 weeks, with two sessions per week and 45-50 min per session. The exercises included both aerobic and strength exercises, considering functional movements and light to moderate intensity. Cognitive stimulation comprehended exercises based on word games, puzzles, mathematical calculations, forward and backward counting, computer exercises, exergames, and games on a balanced platform. Physical assessments (weight, height, and body mass index), health and functional parameters (fitness tests: chair stand, arm curls, chair sit-and-reach, eight feet up-and-go, back scratch, 6-min walking, feet together, semi-tandem, and full tandem), lipid profile (total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and triglycerides), measures of lipid peroxidation damage, thiobarbituric acid reactive substances (TBARS), total antioxidant capacity (TAC), and BDNF were measured in plasma, based on which analyses were performed before and after the 24 weeks of the multicomponent exercise intervention. The results showed an overall improvement in physical and functional performance. Regarding biochemical measures, multicomponent exercises lead to a significant decrease in oxidative damage. The results indicate that multicomponent exercise training induces benefits in functional capacity and reduces damage due to oxidative stress.

Mitochondria research and neurodegenerative diseases: On the track to understanding the biological world of high complexity.

From the simple unicellular eukaryote to the highly complex multicellular organism like Human, mitochondrion emerges as a ubiquitous player to ensure the organism's functionality. It is popularly known as "the powerhouse of the cell" by its key role in ATP generation. However, our understanding of the physiological relevance of mitochondria is being challenged by data obtained in different fields. In this review, a short history of the mitochondria research field is presented, stressing the findings and questions that allowed the knowledge advances, and put mitochondrion as the main player of safeguarding organism life as well as a key to solve the puzzle of the neurodegenerative diseases.

An Integrative Approach to Characterize the Early Phases of Dimethylhydrazine-Induced Colorectal Carcinogenesis in the Rat.

This study aimed to characterize an animal model of colorectal cancer (CRC) in the early stages of disease development. Twenty-nine male Wistar rats were divided into two control groups (CTRL1 and CTRL2), receiving EDTA-saline injections and two induced groups (CRC1 and CRC2), receiving 1,2-dimethylhydrazine (DMH) injections for seven consecutive weeks. CRC1 and CTRL1 were euthanized at the 11th week, while CRC2 and CTRL2 were euthanized at the 17th week. DMH treatment decreased microhematocrit values and IL-6, ghrelin, and myostatin serum levels. Histopathological analysis of intestinal sections showed that DMH-treated rats were characterized by moderate to severe epithelial dysplasia. An adenoma was observed in one animal (CRC2 group), and the presence of inflammatory infiltrate at the intestinal level was primarily observed in DMH-treated animals. DMH also induced Ki-67 immunoexpression. The gut microbiota analysis showed a higher abundance of Firmicutes, Clostridia, Clostridiales, Peptostreptococcaceae, Blautia, Romboutsia, and Clostridium sensu stricto in CRC than CTRL rats, whereas Prevotellaceae, Prevotella, Akkermansia, and Lactobacillus levels were more prevalent in CTRL animals. Our results suggest that this model could be helpful to investigate chemoprevention in the early stages of CRC.

Sperm DNA damage and seminal antioxidant activity in subfertile men.

Supraphysiological ROS levels can lead to apoptosis, lipid peroxidation, and DNA and protein damage. This pilot study aimed to investigate the sperm oxidative damage in subfertile men, to describe the relationship between the antioxidant system and ROS. Sixty-four semen samples were categorised according to the evaluated routine parameters (WHO, WHO laboratory manual for the examination and processing of human semen, 2010). Results were cross-referenced with the DNA damage [Comet (n = 53) and TUNEL (n = 49) assays], antioxidant enzyme activity [SOD (n = 51), CAT (n = 48) and GST (n = 48)], and content of total thiols (n = 36), lipid hydroperoxides (n = 35) and MDA (n = 31). Compared to pathospermic samples, normozoospermic presented 40%-45% fewer spermatozoa with fragmented DNA, 19% fewer hydroperoxides, and slightly higher total thiols and MDA levels. Asthenozoospermic/asthenoteratozoospermic samples had the lowest GST activity. SOD and CAT showed a similar trend. Our results evidenced significant positive correlations between DNA damage and immotile spermatozoa; SOD and CAT, GST and total thiols; CAT and GST; total thiols and sperm concentration; and MDA levels and head/midpiece abnormalities and hydroperoxides. This work contributes to the existing body of knowledge by showing that the oxidative status correlates with the classic sperm analysis parameters. Oxidative stress and DNA damage evaluation might be a valuable diagnostic and prognostic tool in cases of idiopathic male subfertility.

Parabens enhance the calcium-dependent testicular mitochondrial permeability transition: Their relevance on the reproductive capacity in male animals.

Parabens, alkyl ester derivatives from p-hydroxybenzoic acid, are extensively used as antimicrobial preservatives. Nonetheless, due to its widespread and massive employment, several studies highlighted the association between parabens and alterations in the reproductive system. This study aimed to relate the adverse effect of the most commonly used parabens in testis mitochondria with male fertility. From all the parabens used, propyl and butyl were the ones that most negatively decreased the respiratory control ratio. In the case of butyl, inhibitions of 20% and 60% were observed, respectively, at the lowest and highest concentration, when compared to the control group. The membrane potential was only significantly affected by propyl (14%) and butyl (31%), and at a concentration of 250 µM. Succinate dehydrogenase, cytochrome c oxidase, and ATPase activities showed a nonsignificant decrease. Cytochrome c reductase, on the other hand, showed statistically significant inhibitions for both propyl (56%) and butylparaben (55%). The susceptibility to the mitochondrial permeability transition pore (MPTP) opening was increased by all parabens, although this increase was markedly significant for propyl and butyl. These results show that the susceptibility of mitochondria to parabens is dependent on the alkyl chain length and parabens hydrophobicity, and the main mitochondrial target is Complex II-III and MPTP. Hence, this study demonstrates the contribution of parabens exposition to the inhibition of testis mitochondrial function and their putative noxious effect on the male reproductive system.

Use of Parabens (Methyl and Butyl) during the Gestation Period: Mitochondrial Bioenergetics of the Testes and Antioxidant Capacity Alterations in Testes and Other Vital Organs of the F1 Generation.

Since the mid-1920s, parabens have been widely used as antimicrobial preservatives in processed foods and beverages, pharmaceuticals, and cosmetic products. Paraben use continues to generate considerable controversy, both in the general population and in the scientific community itself. The primary purpose of our study was to determine whether parabens (methyl and butyl at concentrations of 100 and 200 mg/kg body weight by subcutaneous injection) during pregnancy of adult female Wistar rats can have an impact on the F1 generation. As far as we know, we are the first to demonstrate that using parabens during pregnancy has negative repercussions on the mitochondrial bioenergetics and antioxidant activity of testicular germ cells in the F1 generation. Our study showed that there was a 48.7 and 59.8% decrease in the respiratory control index with 100 and 200 mg/kg of butylparaben, respectively. Cytochrome c oxidase activity was significantly inhibited (45 and 51%) in both groups. In addition, 200 mg/kg butylparaben promoted a marked decrease in citrate synthase activity, indicating that mitochondrial content decreased in the germ cells, especially spermatocytes and spermatids. Mitochondrial ROS production increased in groups exposed to parabens in a concentration-dependent manner, especially the butyl one (102 and 130%). The groups exposed to butylparaben showed an increase in superoxide dismutase (SOD) and catalase (CAT) activities, while glutathione reductase (GR) and glutathione S-transferase (GST) decreased. With methylparaben, only differences in SOD and GR were observed; for the latter, this only occurred with the highest concentration. The glutathione (GSH)/glutathione disulfide (GSSG) ratio did not undergo any significant change. However, there was a considerable increase in hydroperoxide content in animals exposed to butylparaben, with 100 and 200 mg/kg resulting in 98.6 and 188% increase, respectively. Furthermore, several other organs also showed alterations in antioxidant capacity due to paraben use. In summary, our study demonstrates that paraben use during pregnancy will cause severe changes in the mitochondrial bioenergetics and antioxidant capacity of testicular germ cells and the antioxidant capacity of several other F1 generation organs.

Regulation of cognition: Validation of a short scale for Portuguese first-year university students / Regulación de la cognición: Validación de una escala corta para estudiantes universitarios portugueses de primer año

The quality of learning in Higher Education is particularly dependent on students' skills in regulating their cognition. This regulation requires cognitive and metacognitive skills as well as motivational dimensions. Due to its relevance in explaining students' academic achievement and developing lifelong learning skills, it´s important to increase research in the area. This study aims to adapt and validate a short version of the Regulation of Cognition of Metacognitive Awareness Inventory to first-year Portuguese university students. A sample of 360 students was considered and was identified a three-dimensional structure (Planning, 4 items; Strategies, 7 items; and Monitoring and evaluation, 7 items) with a second-order factor (Regulation of Cognition). The internal consistency values of the reduced scale are within the acceptable parameters for a self-report scale and the correlations with academic achievement at the end of the first year of the university guarantee the predictive validity of the scale. This short version of regulation of cognition measure allows its use in research with other instruments in larger studies and can function as a diagnostic / screening tool to help students in higher education learning challenges

La calidad del aprendizaje en la Educación Superior depende, especialmente, de las habilidades de los estudiantes para regular su cognición. Esta regulación requiere habilidades cognitivas y metacognitivas, así como dimensiones motivacionales. Dada su relevancia en el rendimiento académico y el desarrollo de habilidades para el aprendizaje a lo largo de la vida, es importante aumentar la investigación en el campo. Este estudio pretende adaptar y validar una versión abreviada de la dimensión Regulación de la Cognición del Metacognitive Awareness Inventory para estudiantes universitarios portugueses de primer año. Se empleó una muestra de 360 estudiantes y se identificó una estructura tridimensional (Planificación, 4 ítems; Estrategias, 7 ítems; y Monitoreo y evaluación, 7 ítems) con un factor de segundo orden (Regulación de la cognición). Los valores de consistencia interna de la escala reducida son aceptables para una escala de auto-informe y las correlaciones con el logro académico al final del primer año de la universidad garantizan su validez predictiva. Esta versión abreviada para medir la regulación de la cognición puede usarse en investigación junto con otros instrumentos en estudios más amplios y puede funcionar como una herramienta de diagnóstico para ayudar a los estudiantes en los desafíos del aprendizaje en la enseñanza superior

Elucidating the mechanisms of action of parecoxib in the MG-63 osteosarcoma cell line.

Different types of tumors often present an overexpression of cyclooxygenase-2. The aim of this study was to evaluate the effects of parecoxib (NSAID, cyclooxygenase-2 selective inhibitor) in the behavior of the human osteosarcoma MG-63 cell line, concerning several biological features. Cells were exposed to several concentrations of parecoxib for 48 hours. Cell viability/proliferation, cyclooxygenase-2 expression, morphologic alterations, membrane integrity, cell cycle evaluation, cell death and genotoxicity were evaluated. When compared with untreated cells, parecoxib led to a marked decrease in cell viability/proliferation, in COX-2 expression and changes in cell morphology, in a concentration-dependent manner. Cell recuperation was observed after incubation with drug-free medium. Parecoxib exposure increased lactate dehydrogenase release, an arrest of the cell cycle at S-phase and G2/M-phase, as well as growth of the sub-G0/G1-fraction and increased DNA damage. Parecoxib led to a slight increase of necrosis regulated cell death in treated cells, and an increase of autophagic vacuoles, in a concentration-dependent manner. In this study, parecoxib showed antitumor effects in the MG-63 human osteosarcoma cells. The potential mechanism was inhibiting cell proliferation and promoting necrosis. These results further suggested that parecoxib might be a potential candidate for in-vivo studies.

The Cyclooxigenase-2 Inhibitor Parecoxib Prevents Epidermal Dysplasia in HPV16-Transgenic Mice: Efficacy and Safety Observations.

Carcinogenesis induced by high-risk human papillomavirus (HPV) involves inflammatory phenomena, partially mediated by cyclooxigenase-2. In pre-clinical models of HPV-induced cancer, cyclooxygenase-2 inhibitors have shown significant efficacy, but also considerable toxicity. This study addresses the chemopreventive effect and hepatic toxicity of a specific cyclooxigensase-2 inhibitor, parecoxib, in HPV16-transgenic mice. Forty-three 20 weeks-old female mice were divided into four groups: I (HPV16-/-, n = 10, parecoxib-treated); II (HPV16-/- n = 11, untreated); III (HPV16+/-, n = 11, parecoxib-treated) and IV (HPV16+/-, n = 11, untreated). Parecoxib (5.0 mg/kg once daily) or vehicle was administered intraperitoneally for 22 consecutive days. Skin lesions were classified histologically. Toxicological endpoints included genotoxic parameters, hepatic oxidative stress, transaminases and histology. Parecoxib completely prevented the onset of epidermal dysplasia in HPV16+/- treated animals (0% versus 64% in HPV16+/- untreated, p = 0.027). Parecoxib decreases lipid peroxidation (LPO) and superoxide dismutase (SOD) activity and increases the GSH:GSSG ratio in HPV16+/- treated animals meaning that oxidative stress is lower. Parecoxib increased genotoxic stress parameters in wild-type and HPV16-transgenic mice, but didn't modify histological or biochemical hepatic parameters. These results indicate that parecoxib has chemopreventive effects against HPV16-induced lesions while maintaining an acceptable toxicological profile in this model.

The Academic Self-Regulation Questionnaire: a study with Portuguese elementary school children.

BACKGROUND: Several studies have focused on the assessment of students' motivation because this construct is very important to understand students' learning and how to enhance it. The Academic Self-Regulation Questionnaire (SRQ-A), based on the self-determination theory is a self-report instrument developed to access the reasons why students do their school work. However, there is no Portuguese version of this questionnaire for late elementary students. The primary goal of this research was to analyze the psychometric properties of a Portuguese version of SRQ-A in the domain of Mathematics with elementary school children. METHODS: Participants were 341 elementary school children ranging from 8 to 11 years old from the third and fourth grades. The Portuguese version of the SRQ-A included 24 items assessing four regulatory styles (external, introjected, identified, and intrinsic) in three behavioral categories (homework, classwork, and answering questions in mathematics lessons). To examine the psychometric properties of the instrument, we conducted an exploratory structural equation modeling (ESEM), measured gender and grade invariance, and calculated internal consistency indexes and temporal stability. RESULTS: ESEM analyses supported the original multidimensional structure of the measure with four regulatory styles using a reduced version of the instrument with 16 items. Correlations between the four regulatory styles revealed a simplex pattern consistent with the continuum of self-determination theory. Results showed adequate internal consistency for all regulatory styles (α ≥ .73; CR ≥ .76) and temporal stability (4-month test-retest ≥ .43). The questionnaire showed measurement and structural invariance across gender and grade. Finally, some gender differences were observed; on average, boys scored higher than girls in external regulation. No differences were observed between grades. CONCLUSIONS: Our findings suggest that the Portuguese version of the SRQ-A has good psychometric properties providing adequate support for its use in educational research on motivational styles, including studies concerning gender and grade differences in self-regulation.

The Academic Self-Regulation Questionnaire: a study with Portuguese elementary school children

Abstract Background: Several studies have focused on the assessment of students' motivation because this construct is very important to understand students' learning and how to enhance it. The Academic Self-Regulation Questionnaire (SRQ-A), based on the self-determination theory is a self-report instrument developed to access the reasons why students do their school work. However, there is no Portuguese version of this questionnaire for late elementary students. The primary goal of this research was to analyze the psychometric properties of a Portuguese version of SRQ-A in the domain of Mathematics with elementary school children. Methods: Participants were 341 elementary school children ranging from 8 to 11 years old from the third and fourth grades. The Portuguese version of the SRQ-A included 24 items assessing four regulatory styles (external, introjected, identified, and intrinsic) in three behavioral categories (homework, classwork, and answering questions in mathematics lessons). To examine the psychometric properties of the instrument, we conducted an exploratory structural equation modeling (ESEM), measured gender and grade invariance, and calculated internal consistency indexes and temporal stability. Results: ESEM analyses supported the original multidimensional structure of the measure with four regulatory styles using a reduced version of the instrument with 16 items. Correlations between the four regulatory styles revealed a simplex pattern consistent with the continuum of self-determination theory. Results showed adequate internal consistency for all regulatory styles (α ≥ .73; CR ≥ .76) and temporal stability (4-month test-retest ≥ .43). The questionnaire showed measurement and structural invariance across gender and grade. Finally, some gender differences were observed; on average, boys scored higher than girls in external regulation. No differences were observed between grades. Conclusions: Our findings suggest that the Portuguese version of the SRQ-A has good psychometric properties providing adequate support for its use in educational research on motivational styles, including studies concerning gender and grade differences in self-regulation.

Biochemical and transcriptional analyses of cadmium-induced mitochondrial dysfunction and oxidative stress in human osteoblasts.

Cadmium (Cd) accumulation is known to occur predominantly in kidney and liver; however, low-level long-term exposure to Cd may also result in bone damage. Few studies have addressed Cd-induced toxicity in osteoblasts, particularly upon cell mitochondrial energy processing and putative associations with oxidative stress in bone. To assess the influence of Cd treatment on mitochondrial function and oxidative status in osteoblast cells, human MG-63 cells were treated with Cd (up to 65 µM) for 24 or 48 h. Intracellular reactive oxygen species (ROS), lipid and protein oxidation and antioxidant defense mechanisms such as total antioxidant activity (TAA) and gene expression of antioxidant enzymes were analyzed. In addition, Cd-induced effects on mitochondrial function were assessed by analyzing the activity of enzymes involved in mitochondrial respiration, membrane potential (ΔΨm), mitochondrial morphology and adenylate energy charge. Treatment with Cd increased oxidative stress, concomitantly with lipid and protein oxidation. Real-time polymerase chain reaction (qRT-PCR) analyses of antioxidant genes catalase (CAT), glutathione peroxidase 1 (GPX1), glutathione S-reductase (GSR), and superoxide dismutase (SOD1 and SOD2) exhibited a trend toward decrease in transcripts in Cd-stressed cells, particularly a downregulation of GSR. Longer treatment with Cd (48 h) resulted in energy charge states significantly below those commonly observed in living cells. Mitochondrial function was affected by ΔΨm reduction. Inhibition of mitochondrial respiratory chain enzymes and citrate synthase also occurred following Cd treatment. In conclusion, Cd induced mitochondrial dysfunction which appeared to be associated with oxidative stress in human osteoblasts.