Konstantinos Feidantsis

Starvation and re-feeding affect Hsp expression, MAPK activation and antioxidant enzymes activity of European sea bass (Dicentrarchus labrax).

In the context of food deprivation in fish (wild and farmed), understanding of cellular responses is necessary in order to develop strategies to minimize stress caused by starvation in the aquaculture section. The present study evaluates the effects of long term starvation (1F-3S: one-month feeding-three-month starvation) and starvation/re-feeding (2S-2F: two-month starvation-two-month re-feeding) compared to the control group (4F-0S: four-month feeding-zero month starvation) on cellular stress response and antioxidant defense in organs, like the intestine, the liver, the red and white muscle of European sea bass Dicentrarchus labrax. Molecular responses were addressed through the expression of Hsp70 and Hsp90, the phosphorylation of stress-activated protein kinases and particularly p38 mitogen-activated protein kinase (p38 MAPK) and the extracellular signal-regulated kinases (ERK-1/2). For the determination of the effect of the oxidative stress caused by food deprivation and/or re-feeding, the (maximum) activities of antioxidant enzymes such as glutathione peroxidise (GPx), catalase (CAT) and superoxide dismutase (SOD) as well as the determination of thiobarbituric acid reactive substances (TBARS) were studied. The experimental feeding trials caused a tissue distinct and differential response on the cellular and antioxidant capacity of sea bass not only during the stressful process of starvation but also in re-feeding. Specifically, the intestine phosphorylation of ERKs and antioxidant enzymatic activities increased in the 2S-2F fish group, while in the 1F-3S group an increase was detected in the levels of the same proteins except for GPx. In the liver and the red muscle of 2S-2F fish, decreased Hsp70 and phosphorylated p38 MAPK levels and increased Hsp90 levels were observed. Additionally, SOD activity decreased in the red muscle of 2S-2F and 1F-3S groups. In the liver and red muscle of 1F-3S group Hsp70 levels increased, while the activation of p38 MAPK in the liver decreased. In the white muscle, Hsp90 levels decreased and the phosphorylation of p38 MAPK increased in both feeding regimes compared to control. In the same tissue, GPx and catalase levels were decreased in 2S-2F regime, while SOD levels were decreased in 1F-3S regime.

Metabolic and molecular stress responses of gilthead seam bream Sparus aurata during exposure to low ambient temperature: an analysis of mechanisms underlying the winter syndrome.

The winter syndrome in the gilthead sea bream Sparus aurata indicates that the species is exposed to critically low temperatures in Mediterranean aquaculture in winter. The present study of metabolic patterns and molecular stress responses during cold exposure was carried out to investigate this “disease”, in light of the recent concept of oxygen and capacity limited thermal tolerance. The metabolic profile of fuel oxidation was examined by determining the activities of the enzymes hexokinase (HK), aldolase (Ald), pyruvate kinase (PK), l-lactate dehydrogenase (l-LDH), citrate synthase (CS), malate dehydrogenase (MDH) and 3-hydroxyacyl CoA dehydrogenase (HOAD) in heart, red and white muscle after exposure to temperatures of 10, 14 and 18°C. Especially, the increase in LDH activity combined with the accumulation of l-lactate in tissues indicates that temperatures below 14°C are critical for Sparus aurata and stimulate the anaerobic component of metabolism. Increase in the activity of HOAD suggests that oxidation of free fatty acids might contribute to ATP turnover at low temperatures. The expression of Hsp70 and Hsp90 in all tissues examined revealed a cellular stress response during cooling below 18°C. In the light of winter temperatures in S. aurata cultures around 10°C, our data suggest that the fish are exposed to stressful conditions at the low end of their thermal tolerance window. These conditions likely impair the aerobic capacity of the fish, compromise the rates of growth and reproduction and may contribute to elicit pathological conditions.

Involvement of p38 MAPK in the Induction of Hsp70 During Acute Thermal Stress in Red Blood Cells of the Gilthead Sea Bream, Sparus aurata

The present study aimed to examine the expression and activation of MAPKs (p38 MAPK, ERK1/2, and JNKs) in red blood cells (RBCs) of the gilthead sea bream, Sparus aurata, during thermal stress and investigate their involvement in the expression of heat shock proteins. The data showed that only p38 MAPK is detected in RBCs of Sparus aurata and it is phosphorylated and activated during exposure to increased temperature. Induction of Hsp70 in thermally stressed RBCs was abolished in the presence of the p38 MAPK inhibitor, SB203580, suggesting the involvement of the kinase in this response. This mechanism might play a cytoprotective role in the RBCs of the gilthead sea bream.

Seasonal variations of anti-/apoptotic and antioxidant proteins in the heart and gastrocnemius muscle of the water frog Pelophylax ridibundus☆

In the present work we investigated the seasonal variations of apoptotic and antioxidant proteins in the heart and gastrocnemius muscle of the amphibian Pelophylax ridibundus. Particularly processes studied included the evaluation of hypoxia through the levels of transcriptional factor Hif-1α, of apoptosis through the determination of Bcl-2 and Bax, ubiquitin conjugates levels and the antioxidant defense through the determination of the activity of enzymes such as superoxide dismutase, catalase and glutathione peroxidase. Due to a general metabolic depression during overwintering, levels of the above mentioned proteins and enzymes are generally retained at low levels of expression and activity in the examined tissues of P. ridibundus. On the other hand recovery from overwintering induces oxidative stress, followed by increased levels of the specific proteins and enzymes. A milder up-regulation of antioxidant enzymes during overwintering probably prepares P. ridibundus for oxidative stress during arousal. The seasonal activation of these mechanisms seems to protect this species from these unfavourable conditions.

Nutritional Routine of Tae Kwon Do Athletes Prior to Competition: What Is the Impact of Weight Control Practices?

ABSTRACT Objective: The purpose of the present study was to investigate and assess the common dietary and weight management strategies of Tae Kwon Do (TKD) athletes prior to national competitions, as well as to examine the relationships between these strategies and body weight reduction and sensation of physical condition. Methods: Sixty (n = 60) TKD athletes, 23 women (19.4 ± 2.9 years) and 37 men (20.4 ± 3.6 years), with at least 12.1 ± 3.1 years of experience, participated in the present study. The athletes recorded their dietary intake and physical activity for 3 training days and on a competition day. Bioelectrical impedance was used for body composition estimation. Results: Male athletes consumed 1918 ± 685 kcal/24 hours and 1974 ± 669 kcal/24 hours on training and competition days, respectively, and women 1814 ± 446 kcal/24 hours and 1700 ± 439 kcal/24 hours. TKD athletes had significant negative energy balance (48.6% ± 17.8% to 60.3% ± 26.9%; p < 0.05), with the majority of macro- and micronutritional elements being lower than the recommended values, with significant differences between them, as well as within groups, between weekdays and weekend days (p < 0.05). Females lost most of their weight 2 weeks before the games (3.50 ± 1.00 kg), and males lost most of their weight 3 weeks before (3.16 ± 2.48 kg). The majority of TKD athletes were guided by their coaches for weight management strategies. No significant correlations were found between any body composition variable, weight loss, and any nutritional intake at any time point (p > 0.05). Conclusions: These data suggest that the methods of TKD athletes for rapid weight loss are guided by unspecialized professionals, leading to significant malnutrition, because certain deficiencies in both macro- and micronutrient content are present, with no guaranteed specific reduction of their body mass.

Seasonal variations of cellular stress response in the heart and gastrocnemius muscle of the water frog (Pelophylax ridibundus)

The present study aimed to investigate the seasonal cellular stress response in the heart and the gastrocnemius muscle of the amphibian Pelophylax ridibundus (former name Rana ridibunda) during an 8 month acclimatization period in the field. Processes studied included heat shock protein expression and protein kinase activation. The cellular stress response was addressed through the expression of Hsp70 and Hsp90 and the phosphorylation of stress-activated protein kinases and particularly p38 mitogen-activated protein kinase (p38 MAPK), the extracellular signal-regulated kinases (ERK-1/2) and c-Jun N-terminal kinases (JNK1/2/3). Due to a general metabolic depression during winter hibernation, the induction of Hsp70 and Hsp90 and the phosphorylation of p38 MAPK, JNKs and ERKs are retained at low levels of expression in the examined tissues of P. ridibundus. Recovery from hibernation induces increased levels of the specific proteins, probably providing stamina to the animals during their arousal.

Seasonal Changes in Metabolism and Cellular Stress Phenomena in the Gilthead Sea Bream (Sparus aurata)

Seasonal temperature changes may take organisms to the upper and lower limit of their thermal range, with respective variations in their biochemical and metabolic profile. To elucidate these traits, we investigated metabolic and antioxidant patterns in tissues of sea bream Sparus aurata during seasonal acclimatization for 1 yr in the field. Metabolic patterns were assessed by determining lactate dehydrogenase, citrate synthase, and β-hydroxyacyl CoA dehydrogenase activities, their kinetic properties and plasma levels of glucose, lactate, and triglycerides and tissue succinate levels. Oxidative stress was assessed by determining antioxidant enzymes superoxide dismutase, catalase, and glutathione reductase activities and levels of thiobarbituric acid reactive substances. Xanthine oxidase (XO) activity was determined as another source of reactive oxygen species (ROS) production. Furthermore, we studied the antiapoptotic protein indicator Bcl-2 and the apoptotic protein indicators Bax, Bad, ubiquitin, and caspase as well as indexes of autophagy (LC3B II/LC3B I and SQSTM1/p62) in the liver and the heart to identify possible relationships between oxidative stress and cell death. The results indicate clear seasonal metabolic patterns involving oxidative stress during summer as well as winter. During cold acclimatization, lipid oxidation is induced, while during increased temperatures, warm-induced metabolic activation and carbohydrate oxidation are observed. Thus, oxidative stress seems to be more prominent during warming because of the increased aerobic metabolism. The seasonal profile of apoptosis and XO as another source of ROS matches the results obtained in the laboratory and are interpreted within the framework of oxygen- and capacity-limited thermal tolerance.