The impact of chasing stress on the metabolism of the Atlantic Ghost Crab Ocypode quadrata (Fabricius, 1787).

Ocypode quadrata, a Ghost crab species found along the western Atlantic coast, is considered a bioindicator of anthropogenic impact on sandy beaches. Ghost Crabbing, a touristic activity in which ghost crabs are chased just for fun, is a potentially threatening activity for this crab. In crustaceans, metabolites such as glucose and lactate, and the gene expression of crustacean hyperglycemic hormone (CHH) and heat shock proteins (HSPs) increase when the animals are exposed to several types of stress, including alterations in temperature, salinity, or exposure to xenobiotics. This work was developed to identify if being chased by humans would affect these markers of stress in this species of crab. The effects of chasing stress on hemolymph and tissue metabolites and the gene expression levels of CHH and HSP70 were investigated. The levels of lactate in the hemolymph of stressed crabs were six times higher than those of control crabs immediately after chasing and decreased progressively during recovery, indicating an active anaerobic metabolism during the stress. On the contrary, glucose levels in the hemolymph of the stressed crabs increased progressively from 30 to 60 min after chasing, indicating an inverse correlation between glucose and lactate and the conversion of lactate to glucose by gluconeogenesis. In stressed crabs, the levels of triglycerides in the hemolymph decreased 30 min after chasing, while the opposite tended to occur in the hepatopancreas, indicating that during recovery, the crabs use triglycerides as energy source to sustain aerobic metabolism. Finally, this study demonstrates that ghost crabs are stressed by minimum human contact and that "ghost crabbing" must not be encouraged as a tourist activity.

Acute effects of a single moderate-intensity exercise bout performed in fast or fed states on cell metabolism and signaling: Comparison between lean and obese rats.

AIMS: Although the benefits of exercise can be potentiated by fasting in healthy subjects, few studies evaluated the effects of this intervention on the metabolism of obese subjects. This study investigated the immediate effects of a single moderate-intensity exercise bout performed in fast or fed states on the metabolism of gastrocnemius and soleus of lean and obese rats. MAIN METHODS: Male rats received a high-fat diet (HFD) for twelve weeks to induce obesity or were fed standard diet (SD). After this period, the animals were subdivided in groups: fed and rest (FER), fed and exercise (30 min treadmill, FEE), 8 h fasted and rest (FAR) and fasted and exercise (FAE). Muscle samples were used to investigate the oxidative capacity and gene expression of AMPK, PGC1α, SIRT1, HSF1 and HSP70. KEY FINDINGS: In relation to lean animals, obese animals' gastrocnemius glycogen decreased 60 %, triglycerides increased 31 %; glucose and alanine oxidation decreased 26 % and 38 %, respectively; in soleus, triglycerides reduced 46 % and glucose oxidation decreased 37 %. Exercise and fasting induced different effects in glycolytic and oxidative muscles of obese rats. In soleus, fasting exercise spared glycogen and increased palmitate oxidation, while in gastrocnemius, glucose oxidation increased. In obese animals' gastrocnemius, AMPK expression decreased 29 % and SIRT1 increased 28 % in relation to lean. The AMPK response was more sensitive to exercise and fasting in lean than obese rats. SIGNIFICANCE: Exercise and fasting induced different effects on the metabolism of glycolytic and oxidative muscles of obese rats that can promote health benefits in these animals.

Female Wistar rats present particular glucose flux when submitted to classic protocols of experimental diabetes.

BACKGROUND: Type 1 diabetes mellitus is a prevalent autoimmune disease worldwide. The knowledge of female particularities in metabolic dysfunction is of fundamental importance, leading to better choices for human therapy candidates. The aim of this study is to investigate the glucose flux peculiarities of female rats submitted to two classic experimental diabetes protocols. METHODS: Female Wistar rats, 60 days old, were used to evaluate biochemical and hormonal serum parameters, in addition to skeletal muscle and liver energy stocks and 14C-glucose and 14C-alanine flux. Two different protocols, multiple (25 mg/kg dose) and single (65 mg/kg dose) intraperitoneal streptozotocin, were compared considering the alterations presented 48 h and 30 days after the drug administration. RESULTS: The results showed few indicators of muscle and liver metabolic imbalance. High-single streptozotocin dose promoted 97% and 41% lower glycogen levels in liver and muscle respectively. Multiple-low streptozotocin dose promoted 63% lower lipid synthesis in liver. After 30 days, diabetic animals presented hyperglycaemia in both protocols, 589.5 (529.3/642.3) mg/dL to high-single dose and 374.2 (339.3/530.6) mg/dL to multiple-low dose. However, they did not present lower insulin levels, alterations on muscle glucose uptake, nor higher hepatic gluconeogenesis. CONCLUSION: In conclusion, this study demonstrates that females, at least Wistar rats, are less responsive to classic diabetes protocols established in literature, so mechanisms of experimental diabetes for females need more investigation. After which, therapeutic candidates should be evaluated in such a way sex bias does not present itself as a factor that hinders reproducibility in human studies.

Effects of tributyltin (TBT) on the intermediate metabolism of the crab Callinectes sapidus.

This study investigated if the exposure to tributyltin (TBT), a chemical used worldwide in boat antifouling paints, could result in metabolic disturbances in the blue crab Callinectes sapidus. After the exposure to TBT 100 or 1000 ng.L-1 for 48 and 96 h, hemolymph and tissues were collected to determine the concentration of metabolites and lipid peroxidation. The levels of glucose, lactate, cholesterol, and triglycerides in the hemolymph were not affected by TBT exposure. Hemolymph protein and heart glycogen increased in the crabs exposed to TBT 1000 for 96 h. Anterior gills protein and lipoperoxidation decreased after 96 h in all groups. These results suggest that C. sapidus can maintain energy homeostasis when challenged by the TBT exposure for 48 h and that metabolic alterations initiate after 96 h.

Metabolic and Molecular Subacute Effects of a Single Moderate-Intensity Exercise Bout, Performed in the Fasted State, in Obese Male Rats.

Introduction and objectives: Obesity represents a major global public health problem. Its etiology is multifactorial and includes poor dietary habits, such as hypercaloric and hyperlipidic diets (HFDs), physical inactivity, and genetic factors. Regular exercise is, per se, a tool for the treatment and prevention of obesity, and recent studies suggest that the beneficial effects of exercise can be potentiated by the fasting state, thus potentially promoting additional effects. Despite the significant number of studies showing results that corroborate such hypothesis, very few have evaluated the effects of fasted-state exercise in overweight/obese populations. Therefore, the aim of this study was to evaluate the subacute effects (12 h after conclusion) of a single moderate-intensity exercise bout, performed in either a fed or an 8 h fasted state, on serum profile, substrate-content and heat shock pathway-related muscle protein immunocontent in obese male rats. Methods: Male Wistar rats received a modified high-fat diet for 12 weeks to induce obesity and insulin resistance. The animals were allocated to four groups: fed rest (FER), fed exercise (FEE), fasted rest (FAR) and fasted exercise (FAE). The exercise protocol was a 30 min session on a treadmill, with an intensity of 60% of VO2max. The duration of the fasting period was 8 h prior to the exercise session. After a 12 h recovery, the animals were killed and metabolic parameters of blood, liver, heart, gastrocnemius and soleus muscles were evaluated, as well as SIRT1 and HSP70 immunocontent in the muscles. Results: HFD induced obesity and insulin resistance. Soleus glycogen concentration decreased in the fasted groups and hepatic glycogen decreased in the fed exercise group. The combination of exercise and fasting promoted a decreased concentration of serum total cholesterol and triglycerides. In the heart, combination fasting plus exercise was able to decrease triglycerides to control levels. In the soleus muscle, both fasting and fasting plus exercise were able to decrease triglyceride concentrations. In addition, heat shock protein 70 and sirtuin 1 immunocontent increased after exercise in the gastrocnemius and soleus muscles. Conclusions: An acute bout of moderate intensity aerobic exercise, when realized in fasting, may induce, in obese rats with metabolic dysfunctions, beneficial adaptations to their health, such as better biochemical and molecular adaptations that last for at least 12 h. Considering the fact that overweight/obese populations present an increased risk of cardiovascular events/diseases, significant reductions in such plasma markers of lipid metabolism are an important achievement for these populations.

Liraglutide treatment counteracts alterations in adipose tissue metabolism induced by orchiectomy in rats.

AIMS: The reduction in androgens serum concentration is a physiological condition that accompanies age advancement but can also occur because of prostate cancer and gender affirming treatment or pathological conditions such as functional hypogonadism. However, androgen deficiency is related to a higher risk of developing metabolic disorders such as obesity and type 2 diabetes mellitus (T2DM). Considering that glucagon-like peptide 1 (GLP1) analogs are increasingly used in the treatment of T2DM, we investigated if liraglutide could also attenuate the metabolic changes caused by orchiectomy in rats. MAIN METHODS: Wistar rats were orchiectomized (ORC), and subdivided in four groups: sham saline, sham liraglutide, ORC saline, and ORC liraglutide. After sixty days, metabolic parameters were evaluated in blood, muscle, liver, brown (BAT) and white adipose tissue (WAT) visceral depots. Glucose utilization, oxidation, and conversion to lipids by de novo lipogenesis, and basal and adrenaline-stimulated lipolysis were evaluated in BAT and WAT depots. KEY FINDINGS: Orchiectomy increased triglyceridemia, BAT and rtWAT weight, and lipolysis and reduced glucose utilization. Liraglutide treatment reversed these effects. SIGNIFICANCE: These results indicate that liraglutide improves triglyceridemia and glucose metabolism in WAT depots, which suggests that it may be a promising therapeutic strategy to handle disruptions in energy metabolism caused by androgen deficiency.

Adipose tissue of female Wistar rats respond to Ilex paraguariensis treatment after ovariectomy surgery.

BACKGROUND AND AIM: Metabolic disturbances are known for their increasing epidemiological importance. Ilex paraguariensis presents a potential option for mitigating lipid metabolism imbalance. However, most of the literature to date has not considered sex bias. This study aimed to evaluate the effect of Ilex paraguariensis on the metabolism of different adipose tissue depots in males and females. EXPERIMENTAL PROCEDURE: After ovariectomy, female Wistar rats received daily treatment with the extract (1 g/kg) for forty-five days. Biochemical serum parameters and tissue metabolism were evaluated. Oxidation, lipogenesis and lipolysis were evaluated in brown, white visceral, retroperitoneal and gonadal adipose tissues. RESULTS AND CONCLUSION: The results showed that treatment with the extract led to a reduced weight gain in ovariectomised females in comparison to control. The triglyceride concentration was decreased in males. Glucose oxidation and lipid synthesis in visceral and retroperitoneal adipose tissues were restored in ovariectomised females after treatment. The response to epinephrine decreased in visceral adipose tissue of control males; however, lipolysis in females did not respond to ovariectomy or treatment. These findings highlight the enormous potential effects of I. paraguariensis on lipid metabolism, modulating lipogenic pathways in females and lipolytic pathways in males. Furthermore, the sex approach applied in this study contributes to more effective screening of the effects of I. paraguariensis bioactive substances.

Lactate Administration Reduces Brain Injury and Ameliorates Behavioral Outcomes Following Neonatal Hypoxia-Ischemia.

Neonatal hypoxic-ischemic encephalopathy is a major cause of mortality and disability in newborns and the only standard approach for treating this condition is therapeutic hypothermia, which shows some limitations. Thus, putative neuroprotective agents have been tested in animal models. The present study evaluated the administration of lactate, a potential energy substrate of the central nervous system (CNS) in an animal model of hypoxia-ischemia (HI), that mimics in neonatal rats the brain damage observed in human newborns. Seven-day-old (P7) male and female Wistar rats underwent permanent common right carotid occlusion combined with an exposition to a hypoxic atmosphere (8% oxygen) for 60 min. Animals were assigned to four experimental groups: HI, HI + LAC, SHAM, SHAM + LAC. Lactate was administered intraperitoneally 30 min and 2 h after hypoxia in HI + LAC and SHAM + LAC groups. HI and SHAM groups received vehicle at the same time points. The volume of brain lesion was evaluated in P9. Animals underwent behavioral assessments: negative geotaxis, righting reflex (P8 and P14), and cylinder test (P20). Lactate administration reduced the volume of brain lesion and improved behavioral parameters after HI in both sexes. Thus, lactate administration could be a neuroprotective strategy for the treatment of neonatal HI, a disorder still affecting a significant percentage of human newborns.

Effects of stanniocalcin 1 hormone on lactate metabolism in rat kidney under fed and fasted conditions.

To test the hypothesis of STC-1 participation in maintenance of glucose homeostasis in fed and fasting (48 h) rats, we investigated that this hormone may be implicated in the regulation of renal gluconeogenesis pathway from lactate and lactate oxidation in renal cortex and medulla. Our results demonstrate the hSTC-1 role on lactate metabolism in the renal cortex and medulla from fed and fasting rats. hSTC-1 increased the gluconeogenesis activity in fed state in renal cortex, and this increase was induced by raise in Pck1 gene expression. In fasting animals hSTC-1 increase the renal medulla gluconeogenesis activity, but Pck1 gene expression was not alter. The stimulatory effect of hSTC-1 on 14C-lactate oxidation occurred only in the renal cortex from fed rats. These findings show the hSTC-1 contribution to lactate homeostasis and supplies glucose to other tissues. This response may represent a strategy of action of STC-1 in response to fasting stress as postulated by different authors. On the other hand, hSTC-1 acts downstream of adenylcyclase pathway, decreasing the gluconeogenesis activity induced by cAMP intracellular increase or stimulating the phosphodiesterase activity in the renal cortex. However, no hSTC-1 effect on 14C-lactate oxidation was found after increase in the intracellular cAMP. The findings also revealed that the renal cortex and medulla respond differently to hSTC-1, possibly due to the higher level of STC-1 gene expression in inner renal medulla than in renal cortex.

Diet composition and long-term starvation do not affect crustacean hyperglycemic hormone (CHH) transcription in the burrowing crab Neohelice granulata (Dana, 1851).

The burrowing crab Neohelice granulata is a key omnivorous species in intertidal areas along the southwestern Atlantic from southern Brazil to northern Argentinean Patagonia. This crab is adapted to starvation and can endure natural periods of food deprivation. The metabolic adjustments during starvation depend on the type of diet the crabs were fed previously. Since eyestalk-crustacean hyperglycemic hormone (CHH) is the principal regulator of glucose homeostasis in decapods, we investigated whether CHH transcription was affected by diet composition and starvation. Crabs were maintained in the laboratory for two weeks and subsequently divided in two groups. One received a high carbohydrate (HC) diet, and the other was fed a high protein (HP) diet. After this period, they were starved for four weeks. The full-length cDNA sequence of N. granulata CHH was determined and aligned with CHH sequences of other crabs. Levels of circulating glucose and glycogen were higher in the hepatopancreas and muscle of the HC-fed group and decreased after starvation. Glucose and glycogen concentrations were not altered by starvation in the HP group. Triglyceride levels within the hemolymph were not altered by diet or starvation. However, triglycerides concentration was higher in the hepatopancreas of HC compared to HP-fed group. Long-term starvation and diet composition did not affect CHH transcription.

Effects of stanniocalcin hormones on rat brown adipose tissue metabolism under fed and fasted conditions.

In this study we determined the effect of fed and fasting (48 h) states on the expression of stanniocalcin-1 (Stc1) and stanniocalcin-2 (Stc2) in rat brown adipose tissue (BAT), as well as the in vitro effects of human stanniocalcin 1 and 2 (hSTC-1 and hSTC-2) hormones on lipid and glucose metabolism. In addition, lactate, glycogen levels and hexokinase (HK) activity were determined. In fasting Stc2 expression increased markedly. The targets of action of hSTC-1 and hSTC-2 were glucose uptake and oxidation as well as glycogen storage, controlling the energetic metabolism in BAT. The reduction in glycogen concentration induced by hSTC-2 in fed state might have deleterious consequences in BAT, such as decreased thermogenic activity, FA esterification and other adipocyte functions. On the other hand, the increase of glucose uptake caused by hSTC-1 of fed rats could play a role as a plasma glucose-clearing hormone in the postprandial period.