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Neuroendocrine neoplasms (NENs) are a diverse group of malignancies with a shared phenotype but varying prognosis and response to current treatments. Based on their morphological features and rate of proliferation, NENs can be classified into two main groups with a distinct clinical behavior and response to treatment: (i) well-differentiated neuroendocrine tumors (NETs) or carcinoids (with a low proliferation rate), and (ii) poorly differentiated small- or large-cell neuroendocrine carcinomas (NECs) (with a high proliferation rate). For certain NENs (such as pancreatic tumors, higher-grade tumors, and those with DNA damage repair defects), chemotherapy is the main therapeutic approach. Among the different chemotherapic agents, cisplatin and carboplatin, in combination with etoposide, have shown the greatest efficacy in treating NECs compared to NETs. The cytotoxic effects of cisplatin and carboplatin are primarily due to their binding to DNA, which interferes with normal DNA transcription and/or replication. Consistent with this, NECs, which often have mutations in pathways involved in DNA repair (such as Rb, MDM2, BRCA, and PTEN), have a high response to platinum-based chemotherapy. Identifying mutations that affect molecular pathways involved in the initiation and progression of NENs can be crucial in predicting the response to platinum chemotherapy. This review aims to highlight targetable mutations that could serve as predictors of therapeutic response to platinum-based chemotherapy in NENs.

Introduction: The prevalence of impostor syndrome among PhD students has increased rapidly in recent years, having very negative effects on their mental and psychological health as well as on their doctoral journey. This exploratory study aims to examine whether there is a causal correlation between impostor syndrome among PhD students and empathy among supervisors., Methods: This study encompasses 562 doctoral students (300 females, 262 males) and 152 Tunisian supervisors (68 females, 84 males)., Results: Employing ANOVA, significant influences on impostor syndrome emerge for gender, marital status, professional status, and the doctoral enrollment level ( p  < 0.001). Concurrently, supervisors' empathy is significantly affected by gender, marital status, and experience ( p  < 0.001). Linear regression establishes a noteworthy negative correlation ( p  = 0.045): a 1-unit increase in empathy correlates with a 0.122-unit decrease in impostor syndrome., Discussion: These findings underscore the intricate relationship between socio-professional factors, empathy, and impostor syndrome within the academic milieu, offering crucial insights for interventions and psychological support. The study aligns with the broader context of understanding mental health challenges in academia, emphasizing the imperative for ongoing support initiatives., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Slimi, Muscella, Marsigliante and Bahloul.)

Platinum-based chemotherapeutic agents are widely used in the treatment of cancer. However, their effectiveness is limited by severe adverse reactions, drug resistance, and poor water solubility. This study focuses on the synthesis and characterization of new water-soluble cationic monofunctional platinum(II) complexes starting from the [PtCl(η 1 -C 2 H 4 OEt)(phen)] (1, phen=1,10-phenanthroline) precursor, specifically [Pt(NH 3 )(η 1 -C 2 H 4 OEt)(phen)]Cl (2), [Pt(1-hexyl-1H-imidazole)(η 1 -C 2 H 4 OEt)(phen)]Cl (3), and [Pt(1-hexyl-1H-benzo[d]imidazole)(η 1 -C 2 H 4 OEt)(phen)]Cl (4), which deviate from traditional requirements for antitumor activity. These complexes were evaluated for their cytotoxic effects in comparison to cisplatin, using immortalized cervical adenocarcinoma cells (HeLa), human renal carcinoma cells (Caki-1), and normal human renal cells (HK-2). While complex 2 showed minimal effects on the cell lines, complexes 3 and 4 demonstrated higher cytotoxicity than cisplatin. Notably, complex 4 displayed the highest cytotoxicity in both cancer and normal cell lines. However, complex 3 exhibited the highest selectivity for renal tumor cells (Caki-1) among the tested complexes, compared to healthy cells (HK-2). This resulted in a significantly higher selectivity than that of cisplatin and complex 4. Therefore, complex 3 shows potential as a leading candidate for the development of a new generation of platinum-based anticancer drugs, utilizing biocompatible imidazole ligands while demonstrating promising anticancer properties., (© 2024 Wiley-VCH GmbH.)

The purpose of this study was to investigate the effect of the adapted basketball cycle on the perceived level of difficulty and physical enjoyment in girls and boys with overweight. Sixty-six students with overweight (aged 16-18 years) were randomly assigned to an experimental group (EG, n = 32 including 20 boys and 12 girls) and a control group (CG, n = 34; including 21 boys and 13 girls). Statistical analysis also showed that the levels of perceived difficulty decreased significantly after of seven-week of the adapted basketball cycle in both boys (Δ% = - 0.27; p < 0.001; and girls (Δ% = - 0.36; p < 0.001). There was no significant difference in students who participated in the traditional basketball cycle. A physical activity enjoyment Scale revealed that in girls, the level of enjoyment increased significantly (Δ% = + 0.27; P < 0.001) after an adapted basketball cycle. There was no significant change in physical enjoyment in boys EG (P = 0.808) and participants in the control groups. These results push us to opt more for adapted and motivational learning situations to make the teaching-learning process better, in students with overweight., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. No funding was received for this work., (© 2024 The Authors.)

Phase I clinical trials represent a critical point in drug development because the investigational medicinal product is being tested in humans for the first time. For this reason, it is essential to evaluate and identify the Maximum Tolerated Dose (MTD) and the safety of the new compound. To mitigate the possible risks associated with drug administration and treatment, the European Competent Authority issued various guidelines to provide provisions and harmonize risk management processes. In the UK and Italy, particular attention should be paid to the Medicines & Healthcare Products Regulatory Agency (MHRA) phase I accreditation scheme and the specific rules set by the Italian Drug Authority through the AIFA Determination no. 809/2015. Both reference documents are based on the concept of quality risk management while conducting phase I clinical studies. Moreover, the AIFA determination outlines specific requirements for those sites that want to conduct non-profit phase I clinical trials. Indeed, the document reports peculiar activities to the "Clinical Trial Quality Team", which is a team that should support the clinical site researchers in designing, starting, performing, and closing non-profit phase I studies. In this paper, we provide a general overview of the main European guidelines concerning the management of risks during phase I trials, focusing on the main peculiarities of the schemes and rules set by the MHRA and AIFA.

This study aimed to explore the effect of the motor game, "Clock Motor Games", on the improvement of "Reading and Recording of Time" (RRT) in children with Grade 1 mathematical learning difficulties (MLDs). A within-school cluster-randomized intervention study was conducted with 232 children (aged 6-7 years) with limited physical education experience (0.7 ± 0.3 years). The participants were divided into two groups: a control group, which received conventional teaching on time without any additional motor activities, and an experimental group, which incorporated the concept of time with the "Clock Motor Game", for 3 weeks. The Clock-Reading Test was administered before the intervention (T 0 ), immediately after each session (T 1 ), and five weeks after the intervention (T 2 ) in both groups. The results demonstrated that the experimental group exhibited significantly greater improvements in RRT performance compared to the control group (U = 4416.5; p < 0.001; r = 0.3; medium effect). Additionally, the experimental group was more likely to show progress and less likely to experience regression or stagnation compared to the control group (25% vs. 38.4%). The findings suggest that practicing "Clock Motor Games" can positively contribute to the RRT ability in children with Grade 1 MLD.

Emotional intelligence (EI) has been recently the main target in research on sports psychology. The objectives of this study were to investigate the relation between self-report measures of emotional intelligence, pre-competitive anxiety, and performance among students during the physical education exam of the high school final year. This cross-sectional study was conducted on a sample of 350 students attending the physical education exam in the year 2021-2022. Analysis of the correlations between the scores of Emotional intelligence and anxiety showed that self-confidence was positively correlated with all dimensions of Emotional intelligence (Beta = 0.524; p  = 0.000). Multivariate linear regression analysis showed that the most related items to performance were self-confidence in a positive direction ( r  = 0.56; p  = 0.000) and negatively with somatic anxiety ( r  = -0, 39; p  = 0.000). Considering the Emotional intelligence subscales, hetero-emotional management was significantly positively associated with performance ( p  = 0.000) however emotional perception was negatively correlated with performance ( p  = 0.003). These results demonstrate the importance of social and emotional learning programs for improving self-confidence and better management of emotions during physical education and sports., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Zoghlami, Hawani, Khiari, Mnedla, Marsigliante, Elloumi and Muscella.)

This study aimed to depict the oxidative status variation in judokas during aerobic-dominant mixed effort (AeDME) and anaerobic-dominant mixed effort (AnDME). It is to be expected that the sporting commitment of Judo is a stimulus of oxidative stress leading to the recruitment of antioxidant responses. Blood samples were collected from 17 athletes at rest, immediately after a training session (AeDME) and after a 5-min bout (AnDME). AeDME and AnDME caused significant increases in malondialdehyde (MDA) (p < 0.01 and p < 0.001 respectively) and glutathione (GSH) (p = 0.018 and p < 0.001 respectively). Blood thiol concentrations decreased following AeDME and AnDME (p < 0.001) whilst catalase decreased significantly after AnDME (p = 0.026) only. Uric acid increased significantly after AnDME than after AeDME (p = 0.047) while, conversely, total bilirubin was higher after AnDME than after AeDME (p = 0.02). We may ultimately summarize that AeDME and AnDME caused oxidative stress, higher in AnDME, and some antioxidant response slightly higher in AnDME compared to AeDME. In sports, monitoring of oxidative stress status is recommended as part of the training regimen., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors. Published by Elsevier Ltd.)

Pancreatic cancer is one of the most lethal malignancies with an increasing incidence and a high mortality rate, due to its rapid progression, invasiveness, and resistance to anticancer therapies. In this work, we evaluated the antiproliferative and antimigratory activities of the two organometallic compounds, [Pt( η 1 -C 2 H 4 -OMe)(DMSO)(phen)]Cl ( 1 ) and [Pt( η 1 -C 2 H 4 -OEt)(DMSO)(phen)]Cl ( 2 ), on three human pancreatic ductal adenocarcinoma cell lines with different sensitivity to cisplatin (Mia PaCa-2, PANC-1, and YAPC). The two cationic analogues showed superimposable antiproliferative effects on the tested cells, without significant differences depending on alkyl chain length (Me or Et). On the other hand, they demonstrated to be more effective than cisplatin, especially on YAPC cancer cells. For the interesting cytotoxic activity observed on YAPC, further biological assays were performed, on this cancer cell line, to evaluate the apoptotic and antimetastatic properties of the considered platinum compounds ( 1 and 2 ). The cytotoxicity of 1 and 2 compounds appeared to be related to their intracellular accumulation, which was much faster than that of cisplatin. Both 1 and 2 compounds significantly induced apoptosis and cell cycle arrest, with a high accumulation of sub-G1 phase cells, compared to cisplatin. Moreover, phenanthroline-containing complexes caused a rapid loss of mitochondria membrane potential, ΔΨ M , if compared to cisplatin, probably due to their cationic and lipophilic properties. On 3D tumor spheroids, 1 and 2 significantly reduced migrated area more than cisplatin, confirming an antimetastatic ability., Competing Interests: The authors declare there are no conflicts of interest., (Copyright © 2023 Erika Stefàno et al.)

The feeling of pleasure during physical education (PE) could increase with physical activity participation and adherence for overweight children. While traditional games are known to have positive benefits on motor skill development, especially for children with poorer motor skills, and on the body mass of children, little is known about overweight children's feelings of enjoyment when playing these games. To identify a program of physical activity appreciated by overweight children, we tested the effect of different activities, namely soccer (SO), shot put (SP), and traditional motor games (TMGs), on the feelings of pleasure for 28 overweight male children (aged 12.11 ± 0.63 years, BMI 26.89 ± 0.15 kg·m -2 ) participating in a 3-month cross-over study. To measure affective responses to exercises, we applied a validated feeling scale. The data were collected during eight PE lessons, which were organized for each cycle taught. Post hoc pairwise comparisons revealed a significant difference (Z = -3.195, p < 0.01) between the mean feeling score reported after the SO cycle (2.48 ± 0.41) and that after the TMGs (3.04 ± 0.32). A similar significant difference (Z = -3.304, p < 0.01) was found between the mean feeling scores reported after the SP cycle (2.27 ± 0.32) and the TMGs (3.04 ± 0.32). There was no significant difference between the mean feeling scores reported after the SO (2.48 ± 0.41) and SP cycles (2.27 ± 0.32). In conclusion, the findings of the present study suggest that TMGs may have beneficial effects on the feelings of pleasure for overweight children. Therefore, TMGs might potentially be considered as an alternative to conventional physical activities.

This study aimed to evaluate AT1-R expression in normal and cancerous human kidneys, how these expressions are modified, and AT1-R functionality. AT-1R mRNA expression, determined by real-time PCR, was detected in all samples. AT-1R mRNA increased in well-differentiated cancer (G1, p < 0.01) and decreased 2.9-fold in undifferentiated cancer (G4, p < 0.001) compared with normal kidney tissues. Immunocytochemistry analysis showed that the AT-1R was expressed in the normal tubular epithelium. The glomerulus was also immunoreactive, and as expected, the smooth muscle cells of the vessel walls also expressed the receptor. A total of 35 out of 42 tumors were AT-1R positive, with the cell tumors showing varying numbers of immunoreactive cells, which were stained in a diffuse cytoplasmic and membranous pattern. Computer-assisted counting of the stained tumor cells showed that the number of AT-1R-positive cells increased in the well-differentiated cancers. The functionality of AT-1R was assessed in primary cultures of kidney epithelial cells obtained from three G3 kidney cancer tissues and corresponding histologically proven non-malignant tissue adjacent to the tumor. Indeed, Ang II stimulated, in a dose-dependent manner, the 24 h proliferation of normal kidney cells and cancer cells in the primary culture and phosphorylated extracellular regulated kinases 1 and 2. In conclusion, Ang II may be involved in the growth or function of neoplastic kidney tissue.

The global prevalence of childhood and adolescent overweight and obesity increases rapidly. Physical activity plays a major role in the prevention of obesity. The present study aimed to analyze the effect of adapted basketball sessions according to the empathic capacity of adolescent girls with overweight. Forty-two girls with overweight (age: 16.09 ± 0.85; years; height: 1.64 ± 0.67 m: weight: 73.02 ± 0.61 kg; BMI: 27.15 ± 1.37) volunteered to participate in the study and were randomly assigned to the experimental group (EG, n  = 21) and control group (CG, n  = 21). EG was submitted to a basketball intervention adapted to students with obesity while the CG performed classic basketball exercises for 7 weeks. Each week girls had 2 basketball teaching-learning sessions, lasting 50 min. The participants' empathy was assessed before and after the intervention using the Favre CEC. The results showed that adaptation intervention was associated with a significant emotional contagion decrease (Δ% = 0.466) and splitting with emotions (Δ% = 0.375), and with an empathy increase (Δ% = 1.387), in EG compared to CG. No significant difference was assessed in the empathy CG, before and after the intervention. This study demonstrated that adapted physical education classes could be an effective strategy to improve empathetic skills and inclusion of overweight girls as well as a means to prevent obesity., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Slimi, Marsigliante, Ciardo, Bahloul, Selmi, Jebabli and Muscella.)

This study aimed to evaluate the effectiveness of physically active breaks of a total duration of 10 min a day, introduced during curricular lessons, together with a 10 min physical activity intervention during the daily school recess period on obesity prevention, fitness, cognitive function, and psychological well-being in school-aged children. A sample of 310 children (139 boys vs. 171 girls), aged between 8 and 10 years (9.82 ± 0.51), was selected. Our strategy was implemented over a 6-month period and the participants were randomly assigned to either the intervention group ( n = 157) or the non-intervention (control) group ( n =153). In the intervention group, a significant decrease ( p < 0.05) in body mass index, waist circumference, waist-height ratio, and relative body fat mass was achieved after the intervention (T1) compared to the values measured before intervention (T0); in the control group, no differences emerged between T0 and T1 for any of the parameters considered. We found a significant increase in the intervention group in standing long jump, Ruffier, and sit and reach test scores ( p < 0.001 for all). At T0, cognitive test scores did not differ between the girls and boys or between the intervention and control groups; instead at T1, significant differences were observed in the two groups regarding the total number of responses and the concentration performance scores ( p < 0.001). Consistently, in the intervention group, well-being levels significantly increased between T0 and T1 ( p < 0.001). Finally, the intervention had significant effects on the children regardless of gender. We may therefore conclude that schools should create more opportunities for teachers and students to introduce intervention strategies to promote regular PA during school recess.

We investigated the effect of the role of the joker in children with obesity (OCs) on integration and physio-psychological responses during small-sided games (SSG) training programs. Sixteen OC students (age 13.8 ± 0.73 years) performed training programs consisting of two sessions a week for three weeks. The experimental protocol consisted of 16 teams of 4 children (3 of normal weight and 1 OC). The 16 teams were divided into 2 groups, one with an OC playing as the joker (SSG-J) and the other group with OC playing as non-joker (SSG-NJ). Maximum heart rates (HRmax), blood lactate concentration [La] and OMNI-Child perceived exertion were measured at the end of each SSG. A physical activity enjoyment Scale (PACES) was accomplished during physical activity for the evaluation of feelings in OCs. Additionally, the profile of mood states (POMS) was measured before and after the SSG-J and SSG-NJ programs. HRmax, [La], perceived exertion, and PACES scores were significantly higher after the SSG-J compared with SSG-NJ (increments of 6.4%, 31.7%, 19.5% and 18.1%, respectively). The score of the POMS variables was positively increased in the presence of jokers. The vigor score increased by 30%, while tension and total mood disturbance scores decreased by 27.6% and 4.5%, respectively. These findings suggest that the joker role could be effective in improving integration, physical enjoyment, physiological responses and mood states in OCs when a team game is used during PE sessions. PE teachers could then program joker exercises with the aim of improving OCs' physical commitment willingness to play.

Background: Phase 1 clinical trials represent a critical phase of drug development because new candidate therapeutic agents are tested for the first time on humans. Therefore, international guidelines and local laws have been released to mitigate and control possible risks for human health in agreement with the declaration of Helsinki and the international Good Clinical Practice principles. Despite numerous scientific works characterizing the registered clinical trials on ClinicalTrials.gov, the main features and trends of registered phase 1 clinical trials in Europe have not been investigated. This study is aimed at assessing the features and the temporal trend of distribution of phase 1 clinical studies, carried out in the five largest European countries over a ten-year period (2012-2021), and to evaluate the impact of the Italian regulatory framework on the activation of such studies., Methods: The main data and characteristics of phase 1 clinical studies registered on the ClinicalTrials.gov database for France, Germany, Italy, Spain and the United Kingdom have been investigated and subsequently compared. The above-mentioned countries were selected based on similarities in terms of demographic and Gross Domestic Product (GDP) data available on official government websites. (3) Results: A total number of 6878 phase 1 clinical trials were registered for the five selected countries in the ClinicalTrials.gov database during the ten years analyzed; the studies were predominantly randomized (39.33%) and for-profit (76.64%). The most represented area of investigations was oncology (52.15%), followed by hematology (24.99%) and immunology (12.04%). The variability observed between the analyzed countries showed that the UK, Germany and France presented the highest reduction in the number of phase 1 clinical trials, while for Spain and Italy, a stable/increased trend was observed, although with a lower number of trials registered on the ClinicalTrials.gov database. (4) Conclusions: Italy displayed the lowest number of registered phase 1 clinical trials, even though it showed a stable trend over the years. In this regard, the Italian regulatory framework must urgently be adapted to that of other European countries (Spain has been the first country to implement the new Regulation (EU) No 536/2014) and streamline the process of clinical trial application to increase the attractiveness of the country. Moreover, nonprofit phase 1 clinical trials (which represent 19.81% of the total number of phase 1 clinical trials registered in Italy vs. 80.19% of profit phase 1 clinical studies) should be promoted and supported by the institutions, even from a financial point of view, to allow independent researchers to develop new therapeutic drugs.

Background : We had previously synthetized a macromolecular prodrug consisting of oxidized Alginate and dopamine (AlgOx-Da) for a potential application in Parkinson disease (PD). Methods : In the present work, we aimed at gaining an insight into the interactions occurring between AlgOx-Da and SH-SY5Y neuronal cell lines in view of further studies oriented towards PD treatment. With the scope of ascertaining changes in the external and internal structure of the cells, multiple methodologies were adopted. Firstly, fluorescently labeled AlgOx-Da conjugate was synthetized in the presence of fluorescein 5(6)-isothiocyanate (FITC), providing FITC-AlgOx-Da, which did not alter SH-SY5Y cell viability according to the sulforhodamine B test. Furthermore, the uptake of FITC-AlgOx-Da by the SH-SY5Y cells was studied using scanning near-field optical microscopy and assessments of cell morphology over time were carried out using atomic force microscopy. Results : Notably, the AFM methodology confirmed that no relevant damage occurred to the neuronal cells. Regarding the effects of DA on the intracellular reactive oxygen species (ROS) production, AlgOx-Da reduced them in comparison to free DA, while AlgOx did almost not influence ROS production. Conclusions : these findings seem promising for designing in vivo studies aiming at administering Oxidized Alginate Dopamine Conjugate for PD treatment.

We investigated the effects of high-intensity interval (HIIT) and small-sided games (SSG) training programs on physiological and psychological responses in young soccer players. Twenty-four male soccer players (age 16.7 ± 0.9 years) were divided into two groups (HIIT and SSG) and completed a 6-week training programs consisting of two training sessions a week. HIIT consisted of intermittent runs lasting 15 s at the 110% maximum aerobic speed followed by a passive recovery lasting 15 s; SSG was instead structured in a 4 versus 4 players games on a playing field of 25 × 35 m. The muscular power of the lower body was assessed before and after each training session using the 5-jump test to leg length, and two incremental field tests (VAMEVAL test and modified agility t -test). Our results show that HIIT and SSG have similar beneficial effects on the variables connected to the soccer-specific performance and the endurance with little influence on neuromuscular performances. Psychological responses were assessed using the "physical activity enjoyment scale", the Hooper index and the POMS variables. Results showed that SSG offers greater perceived enjoyment than what can be achieved with HIIT. In conclusion, SSG can be considered an effective, more motivating, and fun soccer-specific training that brings about physiological responses and neuromuscular adjustments in young soccer players.

Childhood obesity represents a serious public health problem and this study evaluates the effectiveness of a 6-month educational intervention on lifestyle, nutrient adequacy, and diet quality in the school setting in improving the knowledge and behavior of primary school children regarding correct eating habits. The strategy was implemented over a 6-month period and participants were randomly assigned to either the intervention group ( n = 200) or the non-intervention group (control group, n = 197). Participants had a mean body mass index of 18.3 ± 2.7 kg/m 2 and its variation in the intervention group (-2.7 ± 0.5 kg/m 2 ) was significantly different from that in the control group (3.41 ± 0.8 kg/m 2 ). In the experimental group, there were significant differences between the proportion of children who were overweight, underweight, normal weight, or obese before and after intervention ( p < 0.05). The best results were seen in the female sex, and after the intervention, there were no more girls with obesity. Furthermore, there were significant waist circumference decrement effects in the intervention group compared to the control group ( p < 0.05). Finally, many of the participating children acquired healthy eating habits. Therefore, the quantitative results obtained suggest that a school intervention program represents an effective strategy to prevent and improve the problem of childhood overweight and obesity., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Marsigliante, Ciardo, Di Maglie, My and Muscella.)

The purpose of this study was to investigate the relationship between mental fatigue induced by a demanding cognitive task and impaired physical performance in endurance due to a higher perception of effort. A total of 12 healthy adults and volunteers, who had previously practiced endurance activities for 4 to 8 h per week, performed a one-hour cognitive task involving either the process of response inhibition (Stroop task) or not (visualization of a documentary as control task), then 20 min of pedaling on a cycle ergometer at a constant perception of effort while cardio-respiratory and neuromuscular functions were measured. The Stroop task induces subjective feelings of mental fatigue (vigor: 3.92 ± 2.61; subjective workload: 58.61 ± 14.57) compared to the control task (vigor: 5.67 ± 3.26; p = 0.04; subjective workload: 32.5 ± 10.1; p = 0.005). This fatigue did not act on the produced perceived effort, self-imposed, and did not affect the cardio-respiratory or neuromuscular functions during the subsequent physical task whose type was medium-term endurance. Regardless of the mental condition, the intensity of physical effort is better controlled when the participants in physical activity control their perception of effort. Mental fatigue does not affect subsequent physical performance but estimated perceived exertion, which increases with the intensity and duration of the exercise.

In order to maximize adaptations and to avoid nonfunctional overreaching syndrome or noncontact injury, coaches in high-performance sports must regularly monitor recovery before and after competitions/important training sessions and maintain well-being status. Therefore, quantifying and evaluating psychometric states of recovery during the season in sports teams such as soccer is important. Over the last years, there has been substantial growth in research related to psychometric states of recovery in soccer. The increase in research on this topic is coincident with the increase in popularity obtained by subjective monitoring of the pre-fatigue state of the players before each training sessions or match with a strong emphasis on the effects of well-being or recovery state. Among the subjective methods for players' control, the Hooper index (HI) assesses the quality of sleep during the previous night, overall stress, fatigue, and delayed-onset muscle soreness. Additionally, the total quality of recovery (TQR) scale measures recovery status. The HI and TQR recorded before each training session or match were affected by the variability of training load (TL) and influenced the physical and technical performances, and the affective aspects of soccer players. Researchers have recommended wellness monitoring soccer players' psychometric state of recovery before each training session or match in order to detect early signs of fatigue and optimize high-level training performance. This method allows for better detecting signs of individual fatigue and allows coaches to adapt and readjust the TL, and avoid physical and technical gaps in order to improve the performance of soccer players.

NMR-based metabolomics is a very effective tool to assess the tumor response to drugs by providing insights for their mode of action. Recently, a novel Pt(II) complex, [Pt(ƞ 1 -C 2 H 4 OMe)(DMSO)(phen)] + (phen =  1,10-phenanthroline), Pt-EtOMeSOphen, was synthesized and studied for its antitumor activity against eight human cancer cell lines. Pt-EtOMeSOphen showed higher cytotoxic effects than cisplatin in most of the cancer cell lines and in particular against the neuroblastoma cell line (SH-SY5Y). In this study, the mechanism of action of Pt-EtOMeSOphen on SH-SY5Y cells was investigated using 1 H NMR-based metabolomics and compared with cisplatin. The observed time response of SH-SY5Y cells under treatment revealed a faster action of Pt-EtOMeSOphen compared with cisplatin, with a response already observed after six hours of exposure, suggesting a cytosolic target. NMR-based metabolomics demonstrated a peculiar alteration of the glutathione metabolism pathway and the diacylglycerol expression., Competing Interests: The authors declare there are no conflicts of interest., (Copyright © 2022 Federica De Castro et al.)

There is very little about the impact that sports training has on female football referees. Therefore, we determined the effects of a 40-week physical preparation, including a full football season, on plasma testosterone and cortisol concentrations and physical performance in female football referees. Plasma cortisol and testosterone concentrations were assayed together with fitness tests at the beginning of the training period (T0, in September), after 8 weeks from T0 (T1), at the mid of the season (T2, 24 weeks after T0), and at the end of the season (T3, in June, 40 weeks after T0). Plasma cortisol increased during the first period and up to T2 (from 15.4 ± 4.7 to 28.5 ± 3.9 µg/dl; p < 0.001), and then decreased at the end of the season (T3: 16.0 ± 2.4 µg/dl). Plasma testosterone concentration in T0 was 14.2±0.37 µg/dl and increased in T1 (57.1 ± 3.7 µg/dl) and T2 (47 ± 3.7 µg/dl) and then decreased in T3 (33.5 ± 2.8 µg/dl). Resting testosterone levels in women were very low (14,2 ± 0.37 µg/dl) (Figure 3c). Testosterone increased in T1 (57.1 ± 3.7 µg/dl) and T2 (47 ± 3.7 µg/dl) whilst, at the end of the season, its concentration decreased (33.5 ± 2.8 µg/dl) (Figure 3c). Significant improvements were observed in all physical performances during the observed period (ANOVA, p < 0.05). Finally, testosterone and cortisol concentrations significantly (p < 0.0001 for both) correlated with maximal oxygen consumption. In T1, testosterone concentration was also significantly correlated with running speed test (p < 0.001). In conclusion, training induces endocrine changes in order to maintain body homeostasis in women referees. It is important that coaches and sports scientists regularly observe changes in endocrine function induced by training and matches in female referees, because they can help maximize referees' performance and limit cases of overtraining., (© 2022 The Authors. Physiological Reports published by Wiley Periodicals LLC on behalf of The Physiological Society and the American Physiological Society.)

A variety of aquatic training regimens have been found to be beneficial for individuals with autism spectrum disorders (ASD) in multiple domains. This study investigated and compared the efficacy of two aquatic training regimens (technical vs. game-based) on gross motor skills, stereotypy behavior and emotion regulation in children with ASD. Twenty-two autistic children were randomly assigned into three groups: two experimental groups performed either a technical aquatic program or a game-based aquatic program and a control group. Participants were assessed before and after an 8-week training period, with the Test of Gross Motor Development, the stereotypy subscale of the Gilliam Autism Rating Scale, and the Emotion Regulation Checklist. A significant effect for time was found in gross motor skills and stereotypy behavior in both experimental groups. An improvement in gross motor skills was observed in both experimental groups compared to the control group. A small pre-post change effect in emotion functioning was found in all groups. No significant differences were observed between the experimental groups in all assessed variables. Our findings provide additional evidence suggesting the effectiveness of beneficial effects of aquatic activities on the motor and social skills that underpin the hypothesis that motor and intellectual domains are highly interrelated in autistic children.

This prospective cohort study aimed to evaluate whether COVID-19 lockdown caused biological, psychological, and/or physical performance variations in footballers. We compared the 2018/2019 and 2019/2020 seasons evaluating the plasma volume, hematological parameters, iron/ferritin, creatine kinase, vitamin D, cortisol, testosterone, and physiological state of players of the Italian football major league (Serie A). Measurements were performed before the preparatory period (T0), at the beginning (T1) and in the middle (T2) of the championship, and in March (T3) and at the end of season (T4). The results showed that in the 2019/2020 season affected by the lockdown, the weight, BMI, and fat mass percentage were higher than in the previous season. Hematocrit, hemoglobin, red blood cells, and ferritin decreased during both seasons, more significantly than in the regular season. During both seasons, creatine kinase increased from T2 whilst iron concentrations decreased in T3. Testosterone increased in both seasons from T0 to T3 and returned to initial levels at T4; cortisol increased in T2 and T3 during the 2018/2019 season but not during the COVID-19 season. Physical performance tests revealed differences associated with lockdown. Thus, although from a medical point of view, none of the evaluated changes between the two seasons were clinically relevant, training at home during lockdown did not allow the players to maintain the jumping power levels typical of a competitive period.

The global prevalence of childhood obesity is high. Obesity main causes are linked to sedentary lifestyles. Increasing physical activity (PA) and reducing sedentary activities are recommended to prevent and treat obesity. This study aimed to evaluate the effectiveness of a 6-month school PA intervention on obesity prevention and healthy behaviors in school-aged children. Participating students (10-11 years of age) were randomly divided into an intervention group and a control group. Children in the intervention group (n = 80) participated in a multicomponent PA that included improvement in extracurricular physical activities (with an additional 40 min per day for 5/6 days per week). Children (n = 80) in the control group participated in usual practice. Participants had mean body mass index of 19.7 ± 2.9 kg/m 2 , and 33.7% of them were overweight or with obesity at T0. The change in body mass index in intervention group (-2.4 ± 0.6 kg/m 2 ) was significantly different from that in control group (3.01 ± 1.8 kg/m 2 ). The effects on waist circumference, waist-to-height ratio, and physic fitness were also significant in intervention group compared with control group (all p < 0.05). Furthermore, there is a significant decrease in overweight or children with obesity in the experimental group (to 17.5%, p < 0.05). These findings suggest that a school-based intervention program represents an effective strategy for decreasing the number of overweight and children with obesity., (© 2022 The Authors. Physiological Reports published by Wiley Periodicals LLC on behalf of The Physiological Society and the American Physiological Society.)

Salvia clandestina L. is a wild perennial species present in the Salento area of Italy. Here, we examined the in vitro effects of an aqueous extract of S. clandestina L. on the MG-63 osteosarcoma cell line. The extract reduced osteosarcoma cell viability mainly by way of apoptosis, as we observed (1) upregulation of gene and protein expression of p53, cyclin-dependent kinase inhibitors p21 WAF1 and p27 Kip1 , and proapoptotic BAX; (2) activation of caspases; and (3) induction of a sub-G 1 peak in the cell cycle. The mitogen-activated protein kinases (MAPKs) JNK1/2 and p38 are activated and involved in the intracellular effects of the S. clandestina extract, as preincubation with the JNK1/2 inhibitor SP600125 or the p38 inhibitor SB203580 significantly decreased S. clandestina extract-induced cytotoxicity and inhibited increase in p53, p21 WAF1 , p27 Kip1 , and BAX. SP600125 also inhibited mRNA levels for all the aforementioned proteins, while SB203580 only affected p53 mRNA. Furthermore, S. clandestina extract treatment counteracted epithelial-to-mesenchymal transition, inhibited cell migration, and decreased the expression and activity of matrix metalloproteinase MMP2. In addition, S. clandestina extract enhanced the cytotoxic activity of cisplatin on MG-63 cells through downregulation of the Akt/PKB protein kinase. We conclude that S. clandestina extract may be a novel agent for osteosarcoma treatment., (© 2021 The Authors. Annals of the New York Academy of Sciences published by Wiley Periodicals LLC on behalf of New York Academy of Sciences.)

As no study has explored the impact of physical stress on hypothalamic-pituitary-gonadal axis hormones over a long period, the purpose of this study was to determine the effects of the football season period on plasma cortisol and testosterone concentrations and referee's physical performances. Physical tests and plasma cortisol and testosterone concentrations were assayed before the beginning of the training period, just after the training period, at the middle of the season, and at the end of the season, in 29 male football referees and 30 healthy control subjects. Results showed significant differences in hormone concentrations at the four-time points evaluated. Plasma cortisol increased during the first training period from 15.8 ± 3.8 to 21.7 ± 5.1 µg/dl (p < 0.001), then decreased during the season and at the end of it was 18.7 ± 2.4 µg/dl. Before the beginning of the training period, plasma testosterone concentration was 386.1 ± 58.8 ng/dl; after the training period, it increased to 572.2 ± 88.1 ng/dl (p < 0.001) and then returned to baseline levels at the end of the season. Between the start of the training period and the end of the season, significant differences were observed in physical performances of referees. Plasma cortisol and testosterone levels significantly (p < 0.0001 for both) correlated with Yo-Yo intermittent recovery test level 1 (YYIRT1) and maximal oxygen consumption (VO 2max ) at the end of the training period. In the middle season, plasma testosterone concentration only significantly (p < 0.0001) correlated with YYIRT1 and VO 2max . These data underline the importance of set up training protocols that present the prospective to favor positive physiological adaptations., (© 2021 The Authors. Physiological Reports published by Wiley Periodicals LLC on behalf of The Physiological Society and the American Physiological Society.)

Blood lipoproteins are formed by various amounts of cholesterol (C), triglycerides (TGs), phospholipids, and apolipoproteins (Apos). ApoA1 is the major structural protein of high-density lipoprotein (HDL), accounting for ~70% of HDL protein, and mediates many of the antiatherogenic functions of HDL. Conversely, ApoB is the predominant low-density lipoprotein (LDL) Apo and is an indicator of circulating LDL, associated with higher coronary heart disease (CHD) risk. Thus, the ratio of ApoB to ApoA1 (ApoB/ApoA1) is used as a surrogate marker of the risk of CHD related to lipoproteins. Elevated or abnormal levels of lipids and/or lipoproteins in the blood are a significant CHD risk factor, and several studies support the idea that aerobic exercise decreases CHD risk by partially lowering serum TG and LDL-cholesterol (LDL-C) levels and increasing HDL-C levels. Exercise also exerts an effect on HDL-C maturation and composition and on reverse C transport from peripheral cells to the liver to favor its catabolism and excretion. This process prevents atherosclerosis, and several studies showed that exercise training increases heart lipid metabolism and protects against cardiovascular disease. In these and other ways, it more and more appears that regular exercise, nutrition, and strategies to modulate lipid profile should be viewed as an integrated whole. The purpose of this review is to assess the effects of endurance training on the nontraditional lipid biomarkers, including ApoB, ApoA1, and ApoB/ApoA1, in CHD risk.

Following peripheral nerve injury, remnant Schwann cells adopt a migratory phenotype and remodel the extracellular matrix allowing axonal regrowth. Although much evidence has demonstrated that TGF-β1 promotes glioma cell motility and induces the expression of extracellular matrix proteins, the effects of TGF-β1 on Schwann cell migration has not yet been studied. We therefore investigated the cellular effects and the signal transduction pathways evoked by TGF-β1 in rattus norvegicus neuronal Schwann RSC96 cell. TGF-β1 significantly increased migration and invasion of Schwann cells assessed by the wound-healing assay and by cell invasion assay. TGF-β1-enhanced migration/invasion was blocked by inhibition of MMP-2 and MMP-9. Consistently, by real-time and western blot analyses, we demonstrated that TGF-β1 increased MMP-2 and MMP-9 mRNA and protein levels. TGF-β1 also increased MMPs activities in cell growth medium, as shown by gelatin zymography. The selective TGF-β Type I receptor inhibitor SB431542 completely abrogated any effects by TGF-β1. Indeed, TGF-β1 Type I receptor activation provoked the cytosol-to-nucleus translocation of SMAD2 and SMAD3. SMAD2 knockdown by siRNA blocked MMP-2 induction and cell migration/invasion due to TGF-β1. TGF-β1 also provoked phosphorylation of MAPKs extracellular regulated kinase 1/2 and JNK1/2. Both MAPKs were upstream to p65/NF-kB inasmuch as both MAPKs' inhibitors PD98059 and SP600125 or their down-regulation by siRNA significantly blocked the TGF-β1-induced nuclear translocation of p65/NF-kB. In addition, p65/NF-κB siRNA knockdown inhibited the effects of TGF-β1 on both MMP-9 and cell migration/invasion. We conclude that TGF-β1 controls RSC96 Schwann cell migration and invasion through MMP-2 and MMP-9 activities. MMP-2 is controlled by SMAD2 whilst MMP-9 is controlled via an ERK1/2-JNK1/2-NF-κB dependent pathway., (© 2019 International Society for Neurochemistry.)

To test the hypothesis that football training would be accompanied by physiological adaptations and hormonal changes, we analyzed the effects of a whole football season on physical fitness and hormonal concentrations in youth football players. Male football players (n = 29, age 16.51 ± 0.7 years) in a regional professional league and male healthy control subjects (n = 30, age 17.1 ± 1 years) participated to the study. Blood cortisol, testosterone, and growth hormone (hGH) concentrations were assayed before the beginning of the training period (T0), just after the training period (T1), at the middle of the season (T2), and at the end of the season (T3). In each period physical tests and anthropometric measurements were also performed. Results showed significant differences in basal values of cortisol, testosterone, and growth hormone (hGH) in the four time points evaluated (P < 0.01). In addition, the concentrations of hGH were higher in the soccer players group than in control subjects (P < 0.001). Between the start of the training period and the end of the football season significant differences were observed in the anthropometric characteristics and in the physical form of the football players. Furthermore, the hormonal status was significantly correlated with the indicators of the lower limb power (squat-jump [SqJ], and counter-movement-jump [CMJ]) and those of aerobic performance (Yo-Yo intermittent recovery test level 1 (YYIRT1) and maximal oxygen consumption (VO 2max )).These data underscore the importance of establishing training protocols that present the potential to promote positive adaptations without, at the same time, provoking overtraining of young players., (© 2019 Wiley Periodicals, Inc.)

Mitochondrial DNA encodes some proteins of the oxidative phosphorylation enzymatic complex, playing an important role in aerobic ATP production; therefore, it can contribute to the ability to respond to endurance exercise training. The accumulation of mitochondrial mutations and the migratory processes of populations have given a great contribution to the development of haplogroups with a different distribution in the world. Several studies have shown the important role of gene polymorphisms in aerobic performance. In this review, some mitochondrial haplogroups and multiple rare alleles were taken into consideration and could be linked to the athlete's physical performance of different ethnic groups., (Copyright © 2019 Elsevier B.V. and Mitochondria Research Society. All rights reserved.)

Malignant pleural mesothelioma (MPM) is an aggressive malignant tumor in which cisplatin therapy is commonly used, although its effectiveness is limited. It follows that research efforts dedicated to identify promising combinations that can synergistically kill cancer cells are needed. Because we recently demonstrated that ADP inhibits the proliferation of ZL55 cells, an MPM-derived cell line obtained from bioptic samples of asbestos-exposed patients. Our objective in this study was to investigate the hypothesis that ADP also potentiates the cytotoxic activity of cisplatin. Results show that in ZL55 cells ADP enhanced (a) the cytotoxicity of cisplatin by 12-fold, (b) the restraint of cell clonogenic potential cisplatin-mediated, and (c) the number of apoptotic cells. Cisplatin, but not ADP, caused caspases activation; nevertheless, poly(ADP-ribose) polymerase-1 was not only cleaved in cisplatin-treated cells but also in cells treated with ADP alone. Furthermore, ADP, but not cisplatin, decreased mTOR and 6SK phosphorylations. Both ADP and cisplatin increased p53 protein, but ADP was also able to enhance p53 messenger RNA. P53 silencing resulted in a very large decrement of cell death induced by ADP or by cisplatin and reverted ADP effects on mTOR/S6K phosphorylation, suggesting that activated p53 may act as a negative regulator of mTOR. Consistently, the inhibition of mTOR by rapamycin also sensitized cells to cisplatin, and the effects of cisplatin plus rapamycin were identical to those obtained with cisplatin plus ADP. These findings suggest that the combination of ADP and cisplatin may be a promising strategy for the clinical treatment of cisplatin-resistant MPM., (© 2018 Wiley Periodicals, Inc.)

Although an association between cancer progression and matrix metalloproteinase (MMP) 2 and MPP9 expression has been known, the expression, nuclear localization, and physiologically controlled activation of these two MMPs have not been investigated in malignant mesothelioma cells. We examined the expression and intracellular localization of MMP2/9 in ZL55 malignant mesothelioma cells, as well as their regulation by ADP. Using real-time PCR, we showed that activation of the P2Y1 receptor by ADP increased the expression of MMP2/9 mRNAs; MMP2/9 collected from conditioned media also showed an increase in activity; and ADP induced the nuclear localization of MMP2/9. The effects of ADP on transcription of the MMPs were due to activation of c-Src, Akt, and NF-κB, while ERK1/2 phosphorylation was needed for the increase in enzymatic activity and the regulation of nuclear import. We also showed that the nuclear localization of MMP2/9 induced by ADP causes the cleavage and inactivation of poly-ADP-ribose polymerase-1. These findings may help to elucidate the mechanisms regulating MMP2/9 activation in ZL55 human epithelioid mesothelioma cells, and perhaps other cells. Therapeutic approaches that promote ADP accumulation in a tumor environment may constitute an effective means to induce anticancer activity., (© 2018 New York Academy of Sciences.)

The novel [Pt( O , O' -acac)(γ-acac)(DMS)], Ptac2S, Pt(II) complex has recently gained increasing attention as a potential anticancer agent for its pharmacological activity shown in different tumor cell lines, studied both in vitro and in vivo. The mechanism of action of Ptac2S, operating on non-genomic targets, is known to be very different from that of cis -[PtCl₂(NH₃)₂], cisplatin, targeting nucleic acids. In this work, we evaluated the cytotoxicity of Ptac2S on the cisplatin resistant Epithelial Ovarian Carcinoma (EOC), SKOV-3 cells, by the MTT assay. A ¹H-NMR metabolomic approach coupled with multivariate statistical analysis was used for the first time for Ptac2S to figure out the biological mechanisms of action of the complex. The metabolic variations of intracellular metabolites and the composition of the corresponding extracellular culture media were compared to those of cisplatin (cells were treated at the IC 50 doses of both drugs). The reported comparative metabolomic analysis revealed a very different metabolic profile between Ptac2S and cisplatin treated samples, thus confirming the different mechanism of action of Ptac2S also in the Epithelial Ovarian Carcinoma (EOC), SKOV-3 cells line. In particular, higher levels of pyruvate were observed in Ptac2S treated, with respect to cisplatin treated, cells (in both aqueous and culture media). In addition, a very different lipid expression resulted after the exposure to the two drugs (Ptac2S and cisplatin). These results suggest a possible explanation for the Ptac2S ability to circumvent cisplatin resistance in SKOV-3 cells.

The relation between the tumor and its microenvironment is one of the most interesting and less understood issues. Recently, we showed a role of CCL20 chemokine in proning the healthy tissue neighboring the tumor to carcinogenesis. Besides, tumor-secreted CCL20 induced proliferation, migration, and EMT of healthy cells. In this context, we have studied here if CCL20 had effects on the migration of cancer cells and the intracellular pathways used in breast epithelial cells in primary culture. Using molecular (siRNA) and pharmacological (inhibitors) techniques, we found multiple signaling kinases to be activated and involved in CCL20-induced tumor breast cell migration. CCL20 provoked a 2.5-fold increase of cell migration and invasion; CCL20 also enhanced MMP- 2 and MMP-9 mRNAs/protein expression and activities. Cell migration and invasiveness due to CCL20 significantly decreased when MMP-2 and MMP-9 were inhibited in CCL20-stimulated cells. CCL20 controlled MMP-2 expression through the JAK2/STAT3 pathway, while the expression of MMP-9 occurred by PKC-α that activated, consequently, c-Src, Akt, and finally NF-kB. These results reveal a role for CCL20 also in tumor breast cell and point to CCL20 as a novel therapeutic target in cancer., (© 2017 Wiley Periodicals, Inc.)

An altered expression of microRNAs (miRNAs) contributes both to the development of cancer and to the progression of the disease. Malignant tumours and tumour cell lines have widespread deregulated expressions of miRNAs compared to normal tissues. In this study, we investigated the expression profiles of 340 mammalian miRNAs in 93 cases of multiform glioblastoma (primary and secondary glioblastoma tumours), by means of DNA microarrays. We show that the expression profiles of 10 miRNAs can distinguish primary from secondary glioblastoma types. Moreover, we found elevated miR-155 levels in primary and secondary glioblastoma tissues as well as in glioblastoma primary cultures. We hypothesised that γ-aminobutyric acid A receptor 1 (GABRA1) is a miR-155 target, and studied the correlation between miR-155 up-regulation and the GABRA1 protein in cultured glioblastoma cells by miRNA silencing. We show that a decrease in miR-155 expression to normal levels restores the expression of GABRA1, making glioblastoma cells sensitive to signals that inhibit cell proliferation mediated by GABRA1. In conclusion, the expression patterns of different miRNAs characterise primary and secondary glioblastomas. The aberrant overexpression of miR-155 contributes to the malignant phenotype of glioblastoma cells removing growth inhibition.

We developed an array-based subtractive hybridization system for one-step high-throughput subtraction. We printed subtractor RNA up to 10.000 times obtaining an excellent contact surface using a little amount of RNA. During hybridization cDNA, common to subtractor and target samples, remains attached to slide immobilized RNA, leaving free in solution target specific cDNA which after retrieval is cloned.

Introduction: TWO TYPES OF GLIOMATOSIS CEREBRI EXIST: Type I and Type II. We report the results of a histological and genetic study of two cases of gliomatosis cerebri Type II, correlating these results with therapy and prognosis., Case Presentation: Two patients, a 52-year-old man (Patient 1) and a 76-year-old man (Patient 2) with gliomatosis cerebri II were admitted to our institution; they underwent surgical treatment and received radiotherapy and chemotherapy. At the 24-month follow-up, Patient 1 was still alive, while Patient 2 had died. The poor prognosis of Patient 2 was underlined by molecular analysis which showed that the angiogenesis related genes VCAM1 and VEGF were overexpressed, reflecting the high degree of neovascularization., Conclusion: Genes involved in drug resistance and metallothioneins were highly expressed in Patient 2 and this, associated with unmethylated O6-methylguanine methyltransferase, can explain the lack of response to chemotherapy.

We demonstrated previously that in serum-starved MCF-7 breast cancer cell line, Ang II increased Na+/K+ATPase activity and activated the protein kinase C zeta (PKC-zeta) (Muscella et al., 2002 J Endocrinol 173:315-323; 2003 J Cell Physiol 197:61-68.). The aim of the present study was to investigate the modulation of the activity of the Na+/K+ATPase by PKC-zeta in MCF-7 cells. Here, using serum-starved MCF-7 cells, we have demonstrated that the effect of Ang II on the Na+/K+ATPase activity was inhibited by a synthetic myristoylated peptide with sequences based on the endogenous PKC-zeta pseudosubstrate region (zeta-PS) and by high doses of GF109203X, inhibitor of PKCs. When MCF-7 cells, grown in 10% fetal bovine serum (FBS), were stimulated with Ang II a dose- and time-dependent inhibition of the Na+/K+ATPase activity was obtained. Under this growth condition we found that mRNAs for AT1, AT2, and for Na+/K+ATPase alpha1 and alpha3 subunits were unchanged; besides both the activity of the Na+/K+ATPase and the level of PKC-zeta also were unaffected by the serum. The atypical PKC-iota level (present in very low abundance in serum-starved MCF-7) was increased and Ang II provoked its translocation from the cytosol to plasma membrane. PKC-zeta was localized to the membrane, and upon Ang II treatment its cellular localization did not change. The Ang II-mediated decrease of the Na+/K+ATPase activity was inhibited by high doses of GF109203X but not by zeta-PS, thus indicating that such effect was not due to PKC-zeta activity. The treatment of cells with PKC-iota antisense oligodeoxynucleotides inhibited the effects of Ang II on the Na+/K+ATPase activity. Additionally, the effect of Ang II on Na+/K+ATPase activity was also blocked by the phosphatidylinositol 3-kinase (PI3K) inhibitors, wortmannin and LY294002, and by the actin depolymerizing agents, cytochalasin D. In conclusion, in MCF-7 cells Ang II modulates the Na+/K+ATPase activity by both atypical PKC-zeta/-iota. The effects of Ang II are opposite depending upon the presence of the serum-sensitive PKC-iota, with the inhibitory effect possibly due to the redistribution of sodium pump from plasma membrane to the inactive intracellular pool., (Copyright 2005 Wiley-Liss, Inc.)

Objective: We recently reported that the sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) 2b is the SERCA form preferentially expressed in rat thyroid. Moreover, SERCA2b expression dramatically decreases in virally transformed, highly tumorigenic, PC Cl3 thyroid cells. These results suggest that, in the thyroid, SERCA2b, in addition to its housekeeping role, is linked to differentiation and is a regulated gene. We therefore sought to study the effect of TSH, the main regulator of thyroid function, on SERCA2b expression and activity., Methods: PC Cl3 cells were hormone starved in low-serum medium and stimulated for long (48 h) or short (1, 2 and 4 h) times. SERCA2b expression and activity were evaluated by Northern and Western blots, Ca2+-ATPase activity and Ca2+ store content., Results: In PC Cl3 cells, SERCA2b mRNA and protein were induced twofold by a 48-h long treatment with TSH. Long-term elevation (48 h) of intracellular cAMP levels, by forskolin or 8-Br-cAMP, had similar effects on SERCA2b mRNA and protein. We also measured Ca2+-ATPase activity and Ca2+ store content. Both long (48 h) and short (0.5-1 h) treatments with TSH, forskolin or 8-Br-cAMP induced a marked increase of SERCA2b activity. This effect was completely abolished by H89, a specific inhibitor of cAMP-dependent protein kinase A (PKA). TSH and 8-Br-cAMP increased Ca2+ store content after both long (48 h) and short (1-2 h) treatments., Conclusions: These data suggested that TSH/cAMP acts as an important regulator of both SERCA2b expression and activity in the thyroid system, through PKA activation.

We examined the signalling pathways responsible for the Ang II induction of growth in MCF-7 human breast cancer cells. Ang II in MCF-7 cells induced: (a) the translocation from the cytosol to membrane and nucleus of atypical protein kinase C-zeta (PKC-zeta) but not of PKC-alpha, -delta, - epsilon and -eta; (b) the expression of c-fos mRNA and protein; (c) the phosphorylation of the extracellular signal-regulated protein kinases 1 and 2 (ERK1/2). All these effects were due to the activation of the Ang II type I receptor (AT1) since they were blocked by the AT1 antagonist losartan. The Ang II-stimulated ERK1/2 phosphorylation was blocked by (a) high doses of staurosporine, inhibitor of PKC-zeta, and by a synthetic myristoylated peptide with sequences based on the endogenous PKC-zeta pseudosubstrate region (zeta-PS); (b) PD098059, a mitogen-activated protein kinase kinase inhibitor (MAPKK/MEK); and, moreover, (c) the inhibitors of phosphoinositide 3-kinases (PI3K), LY294002 and wortmannin, thus indicating that PI3K may act upstream of ERK1/2. The Ang II-evoked c-fos induction was blocked only by high doses of staurosporine and by zeta-PS whilst PD098059, LY294002 and wortmannin were ineffective, thus indicating that c-fos induction is not due to ERK1/2 activity. When the epidermal growth factor-receptor (EGFR) tyrosine kinase activity was inhibited by the use of its inhibitor AG1478, Ang II was still able to induce ERK1/2 phosphorylation and c-fos expression, therefore proving that the transactivation of EGFR was not required for these Ang II effects in MCF-7 cells. The previously reported proliferation of MCF-7 cells induced by Ang II was blocked by PD098059 and by wortmannin in a dose-dependent manner, thereby indicating that in MCF-7 cells the PI3K and ERK pathways mediate the mitogenic signalling of AT1. Our results suggest that in MCF-7 cells Ang II activates multiple signalling pathways involving PKC-zeta, PI3K and MAPK; of these pathways only PKC-zeta appears responsible for the induction of c-fos., (Copyright 2003 Wiley-Liss, Inc.)

It was previously shown that in rat thyroid PC-Cl3 cell line, a purinergic P2Y receptor increases the concentration of free cytosolic Ca(2+) ([Ca(2+)](i)) via phospholipase C activation. We here studied whether in a transformed cell line (PC-E1Araf) derived from parental PC-Cl3 cells, ATP is still able to transduce the [Ca(2+)](i)-based intracellular signal.We demonstrate the expression of mRNA for P2Y2 in both PC-Cl3 and PC-E1Araf cells; mRNAs for P2Y1, P2Y4, P2Y6 and P2Y11 were absent. In both cell lines activation of P2Y2 receptor provokes a transient increase in [Ca(2+)](i) followed by a lower sustained phase persisting for over 5min in PC-Cl3 and only 1.5 min in PC-E1Araf cells. In both cell lines the [Ca(2+)](i) reached a plateau level significantly higher than the basal [Ca(2+)](i) level persisting for over 10 min. Removal of extracellular Ca(2+) reduced the initial transient response to ATP in PC-Cl3, but not in PC-E1Araf cells, and completely abolished the plateau phase in both cell lines. In the presence of extracellular Ca(2+) thapsigargin (TG) caused a rise in [Ca(2+)](i) significantly higher in PC-Cl3 than transformed PC-E1Araf cells, while in Ca(2+)-free medium the effect of TG was similar in both cell lines. The capacitative Ca(2+)-entry in PC-Cl3 resulted significantly higher than in PC-E1Araf cells. Further studies were performed in order to investigate whether the different effects of ATP on [Ca(2+)](i) was due to variation in divalent cation plasma membrane permeability. PC-E1Araf cells showed a much lower permeability to Ca(2+), Ba(2+), Sr(2+), Mn(2+), and Co(2+) that may be responsible for the differences in purinergic Ca(2+) signaling pathway with respect to parental PC-Cl3 cells.