Plasma Ion Levels Research Articles

Salinity induced changes in the progression of water, ion and nutrient fluxes along the gastrointestinal tract of Atlantic salmon smolt (Salmo salar)

Water ingestion in fish increases with both water salinity and feeding. However, it is unclear whether, during feeding, water ingestion is intended to aid chyme liquefaction in the stomach or to maintain the osmotic homeostasis within the body of the fish. We investigated the effects of increasing water salinity (0, 10, 20, 35 ppt) on the progression of water, ion and nutrient fluxes in the gastrointestinal tract of Atlantic salmon smolt (Salmo salar) fed a commercial-like diet. Furthermore, the effect of water salinity on blood pH, plasma osmolality and ions was investigated. The experiment lasted for 8 weeks. Chyme was collected from 4 gastrointestinal tract (GIT) segments (stomach, proximal, middle and distal intestine) and analysed for dry matter, pH, osmolality, crude protein and mineral content. Water and electrolyte fluxes, kinetic of digestion and faecal digestibility were measured using yttrium oxide (Y2O3) as an inert marker. We found that between 0 and 35 ppt chyme dry matter decreased by 1.6% and 4.8% in the stomach and proximal intestine, respectively. Chyme pH was not affected by water salinity in the stomach, but it increased linearly (p < 0.001) with salinity in all intestinal segments. Chyme osmolality increased linearly (p < 0.001) with salinity in the stomach and it decreased in all intestinal segments. Water fluxes were similar among salinities in the stomach, but they increased nearly fivefold (6.2 versus 27.3 ml g−1 ingested DM) in the proximal intestine between 0 ppt and 35 ppt. An efflux of monovalent ions (Na+ and K+) increased linearly (p < 0.001) with salinity in the proximal intestine. An efflux of divalent ions (Ca2+ and Mg2+) increased curvilinearly (p < 0.001) with salinity in the middle intestine. Plasma osmolality and ion levels increased with salinity. Crude protein digestibility and protease activity decreased significantly with water salinity in the intestine. Our study highlights that when Atlantic salmon moves from freshwater to higher water salinity environments, drinking of saltwater does not interfere with hydration of feed in the stomach, but instead bypasses to the proximal intestine to aid in osmoregulatory water uptake. Therefore, we suggest that water ingestion in seawater fish is intended for osmoregulation rather than to aid digestion by liquefying chyme in the stomach.

Nitric Oxide Mobilizes Intracellular Zn2+ via the GC/cGMP/PKG Signaling Pathway and Stimulates Adipocyte Differentiation.

Plasma and tissue zinc ion levels are associated with the development of obesity. Previous studies have suggested that zinc ions may regulate adipocyte metabolism and that nitric oxide (NO) plays a pivotal role in the regulation of adipocyte physiology. Our previous study showed that chronic NO deficiency causes a significant decrease in adipose tissue mass in rats. Studies also suggested that zinc ions play an important modulatory role in regulating NO function. This study aims to explore the role of zinc ions in NO-regulated adipocyte differentiation. We hypothesized that NO could increase intracellular Zn2+ level and then stimulate adipocyte differentiation. ZnCl2 and the NO donor, NONOate, were used to explore the effects of Zn2+ and NO on adipocyte differentiation. Regulatory mechanisms of NO on intracellular Zn2+ mobilization were determined by detection. Then, Zn2+-selective chelator TPEN was used to clarify the role of intracellular Zn2+ on NO-regulated adipocyte differentiation. Furthermore, the relationship between adipocyte size, Zn2+ level, and NOS expression in human subcutaneous fat tissue was elucidated. Results showed that both ZnCl2 and NO stimulated adipocyte differentiation in a dose-dependent manner. NO stimulated intracellular Zn2+ mobilization in adipocytes through the guanylate cyclase (GC)/cyclic guanosine monophosphate (cGMP)/protein kinase G (PKG) pathway, and NO-stimulated adipocyte differentiation was Zn2+-dependent. In human subcutaneous adipose tissue, adipocyte size was negatively correlated with expression of eNOS. In conclusion, NO treatment stimulates intracellular Zn2+ mobilization through the GC/cGMP/PKG pathway, subsequently stimulating adipocyte differentiation.

Transepithelial potential remains indicative of major ion toxicity in rainbow trout (Oncorhynchus mykiss) after 4-day pre-exposure to major salts.

The Multi-Ion Toxicity (MIT) Model uses electrochemical theory to predict the transepithelial potential (TEP) across the gills as an index of major ion toxicity in freshwater animals. The goal is to determine environmental criteria that will be protective of aquatic organisms exposed to salt pollution. In recent studies, TEP disturbances above baseline (ΔTEP) during short-term exposures to major ions have been proven as indicative of their toxicity to fish, in accord with the MIT model. However, the acute 1-h exposures used in these previous studies might not be realistic relative to the 24h or 96h test periods used for toxicity assessment. To address this temporal inconsistency, the current study investigated both the TEP responses to serial concentrations of 10 major salts (NaCl, Na2SO4, NaHCO3, KCl, K2SO4, KHCO3, CaCl2, CaSO4, MgCl2, MgSO4) and plasma ion levels in juvenile rainbow trout after they had been pre-exposed to 50% of the 96h-LC50 levels of these same salts for 4 days. The pre-exposures caused no mortalities. In general, plasma ions (Na+, K+, Ca2+, Mg2+, Cl-) were well-regulated; however, pre-exposure to sulfate salts resulted in the greatest number of alterations in plasma ion levels. TEP responses remained largely similar to those of naïve trout (without salt pre-exposure). All salts caused hyperbolic concentration-dependent increases in TEP that were well-described by the Michaelis-Menten equation. In the pre-exposed trout, the variation of ∆TEP at the 96h-LC50 concentrations was only 2.2-fold, compared to nearly 28-fold variation among the molar concentrations of the various salts at the 96h-LC50s, identical to the conclusion for naïve trout. Overall, the results remove the temporal inconsistency of previous tests and remain supportive of the MIT model. In addition, the recorded alterations in certain plasma ions, baseline TEP, and Michaelis-Menten constants improve our knowledge on specific physiological responses after extended major ion exposure.

Physiology and aquaculture: A review of ion and acid‐base regulation by the gills of fishes

AbstractFish aquaculture facilities strive to maintain abiotic conditions that optimise somatic growth. Two physiological systems that are critically linked to environmental abiotic factors are ion and acid‐base regulation. Given the detrimental effects of ionic and acid‐base imbalances coupled with the potential energetic costs of ion and acid‐base regulation, it is important to understand how changes in abiotic variables such as salinity, temperature and pH alter ionoregulatory and acid‐base physiology. The gill is the predominant site of ion and acid‐base regulation, two linked processes that are vital to the health of fishes. Most species maintain ion levels in the blood plasma within a narrow range, independent of external ion concentrations. In freshwater (FW), fishes must actively take up ions from their dilute environment to counteract passive ion losses, while in seawater (SW), ion excretion serves to counteract passive salt gain. Ion uptake (FW) and excretion (SW) are coordinated by different subtypes of ion‐transporting cells (ionocytes) expressed in the gill epithelium. In this review, we discuss the function of FW and SW ionocytes in the movement of ions across the gill epithelium and address other features of the gill that are integral to ionoregulation. We also discuss the mechanisms of acid‐base regulation by the gill, which is intimately related to ion regulation because it is coordinated by Na+‐ and Cl−‐linked fluxes of acid (H+) and base () equivalents. These fundamental physiological principles are integral in understanding how fishes in aquaculture respond to relevant disturbances that may occur under culture conditions.

High-fat diet-induced aggravation of cardiovascular impairment in permethrin-treated Wistar rats

This study characterized the impact of post-weaning high-fat diet (HFD) and/or permethrin (PER) treatment on heart dysfunction and fibrosis, as well as atherogenic risk, in rats by investigating interactions between HFD and PER. Our results revealed that HFD and/or PER induced remarkable cardiotoxicity by promoting cardiac injury, biomarker leakage into the plasma and altering heart rate and electrocardiogram pattern, as well as plasma ion levels. HFD and/or PER increased plasma total cholesterol, triacylglycerols, and low-density lipoprotein (LDL) cholesterol levels but significantly reduced high-density lipoprotein (HDL) cholesterol. Cardiac content of peroxidation malonaldehyde, protein carbonyls, and reactive oxygen species were remarkably elevated, while glutathione levels and superoxide dismutase, catalase and glutathione peroxidase activities were inhibited in animals receiving a HFD and/or PER. Furthermore, cardiac DNA fragmentation and upregulation of Bax and caspase-3 gene expression supported the ability of HFD and/or PER to induce apoptosis and inflammation in rat hearts. High cardiac TGF-β1 expression explained the profibrotic effects of PER either with the standard diet or HFD. Masson’s Trichrome staining clearly demonstrated that HFD and PER could cause cardiac fibrosis. Additionally, increased oxidized LDL and the presence of several lipid droplets in arterial tissues highlighted the atherogenic effects of HFD and/or PER in rats. Such PER-induced cardiac and vascular dysfunctions were aggravated by and associated with a HFD, implying that obese individuals may be more vulnerable to PER exposure. Collectively, post-weaning exposure to HFD and/or PER may promote heart failure and fibrosis, demonstrating the pleiotropic effects of exposure to environmental factors early in life.

Differential modulation of oxidative stress, antioxidant defense, histomorphology, ion-regulation and growth marker gene expression in goldfish (Carassius auratus) following exposure to different dose of virgin microplastics

Goldfish (Carassius auratus) juveniles were exposed to virgin polyvinyl chloride microplastics (PVC-MPs) in triplicate at 0, 0.1 or 0.5 mg/L for four days. Afterwards, the histopathology of the gills, liver and intestines were examined, along with various antioxidant enzymes and indicators of oxidative damage (malondialdehyde (MDA) and hydrogen peroxide (H2O2)), in the brain, liver and gills. In addition, we also studied the expression of hepatic insulin-like growth factor-1 (IGF-1), insulin-like growth factor binding protein 1 (IGFBP-1) and growth hormone (GH) receptor, while cortisol receptor (CR) and cytochrome P450 1A (CYP1A) gene expression were assayed in both the liver and gills. Histological analysis revealed PVC-MPs in the intestines at 0.1 and 0.5 mg/L, along with substantially shorter villi. The gills appeared undamaged by PVC-MPs exposure and had limited or no effect to antioxidant activity, Na+/K+-ATPase and H+-ATPase activity or plasma ion levels, but there was a prominent upsurge of the detoxification enzymes glutatione S-transferase (GST) activity and CYP1A expression. Livers showed inflammation and some occurrences of hemorrhaging and necrosis at 0.5 mg/L. While the brain showed some evidence of oxidative damage, the liver was the most susceptible to oxidative damage, based on increased MDA, H2O2 and various antioxidant enzymes. Hepatic expression of IGFBP-1 and GH receptor were significantly downregulated at 0.5 mg/L while CR was upregulated. Results indicate that exposure to environmentally relevant PVC-MP can cause oxidative damage in the brain and liver, adverse histomorphological changes to the intestine and liver and alter the gene expression in goldfish.

Loss of sodium chloride co-transporter impairs the outgrowth of the renal distal convoluted tubule during renal development.

Loss-of-function mutations in the sodium chloride (NaCl) co-transporter (NCC) of the renal distal convoluted tubule (DCT) cause Gitelman syndrome with hypokalemic alkalosis, hypomagnesemia and hypocalciuria. Since Gitelman patients are usually diagnosed around adolescence, we tested the idea that a progressive regression of the DCT explains the late clinical onset of the syndrome. NCC wild-type and knockout (ko) mice were studied at Days 1, 4 and 10 and 6 weeks after birth using blood plasma analysis and morphological and biochemical methods. Plasma aldosterone levels and renal renin messenger RNA expression were elevated in NCC ko mice during the first days of life. In contrast, plasma ion levels did not differ between genotypes at age 10 days, but a significant hypomagnesemia was observed in NCC ko mice at 6 weeks. Immunofluorescent detection of parvalbumin (an early DCT marker) revealed that the fractional cortical volume of the early DCT is similar for mice of both genotypes at Day 4, but is significantly lower at Day 10 and is almost zero at 6 weeks in NCC ko mice. The DCT atrophy correlates with a marked reduction in the abundance of the DCT-specific Mg2+ channel TRPM6 (transient receptor potential cation channel subfamily M member 6) and an increased proteolytic activation of the epithelial Na+ channel (ENaC). After an initial outgrowth, DCT development lags behind in NCC ko mice. The impaired DCT development associates at Day 1 and Day 10 with elevated renal renin and plasma aldosterone levels and activation of ENaC, respectively, suggesting that Gitelman syndrome might be present much earlier in life than is usually expected. Despite an early downregulation of TRPM6, hypomagnesemia is a rather late symptom.

Do Frogs Infected with Batrachochytrium dendrobatidis Avoid Water While Sloughing?

Chytridiomycosis is a skin infection caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd), that has been responsible for amphibian declines and extinctions worldwide. Chytridiomycosis increases the permeability of amphibian skin leading to low plasma ion levels and loss of physiological homeostasis. Because temporary increases in cutaneous salt loss and water uptake occur during sloughing, potentially exacerbating the effects of Bd infection, avoiding water while sloughing might help to alleviate these effects, and could contribute to intra- and interspecific differences in pathophysiology. Furthermore, because the amount of time an infected individual spends in contact with water could affect the severity of ion loss, we questioned whether amphibian behavior is altered in response to infection with Bd. This project utilized continuous infrared video footage of captive Australian Green Tree Frogs (Litoria caerulea) as a method of behavioral analysis, to determine whether infection with Bd altered the duration of water contact, activity levels, and the location of sloughing events. We found that although frogs became less active once infected, there was no change in the duration of time spent in contact with water. Infected frogs showed a decrease in the proportion of sloughing events occurring in contact with water as the experiment progressed. Our study is novel in that it provides insight into the behavioral effects of chytridiomycosis, demonstrating that Bd infection can induce behaviors that might alter disease progression.

A novel functional role of nickel in sperm motility and eukaryotic cell growth

BackgroundMetal ions are essential for numerous life processes. This study aims to investigate the relationship between seminal quality and ion levels in seminal plasma. Basic proceduresA total of 205 semen samples were collected and seminal plasma ion levels were examined with inductively-coupled plasma-mass spectrometry. The nickel function was demonstrated by in vitro assay and cell growth. Main findingsThe low sperm motility group showed distinctively reduced nickel concentration in seminal plasma compared with the normal sperm motility group. However, arsenic, sulfur, selenium, magnesium and zinc were negatively associated with sperm quality. No significant relationship between other examined cations and semen quality was observed. In vitro assay suggested low concentration of nickel significantly increased sperm total motility and progressive motility. Cell growth assay further confirmed nickel promoted eukaryotic yeast cell growth. Nickel level in seminal plasma may play important functions to determine sperm quality. Principal conclusionsOur study reveals a strong correlation between S, Mg, Se, Zn, As, Ni and seminal quality as well as discovers a novel functional role of nickel in sperm motility and eukaryotic cell growth. These findings may provide a potential avenue for assessment of sperm quality and treatment of reproduction disorders.

Lack of change in swimming capacity (Ucrit) following acute salinity exposure in juvenile shortnose sturgeon (Acipenser brevirostrum).

The Saint John River (SJR) is home to the only Canadian population of shortnose sturgeon, Acipenser brevirostrum. Adult shortnose sturgeon routinely enter saltwater to forage, yet less is known about how juveniles cope with the associated osmoregulatory pressures. Recently, it has been shown that short-term (24h) exposure to saltwater causes significant changes to ion and water levels in juvenile shortnose. In some species of fish, notably salmonids, it has been shown that shifts in fluid and ion levels following saltwater challenges reduce the swimming capacity. The relationship between ion concentration and swimming capacity is not well understood for sturgeon species. Our research aimed to determine whether short-term salt exposure affects swimming ability in juvenile shortnose sturgeon. Juvenile, SJR, hatchery-raised shortnose sturgeon (< 1year old) were exposed to salinities of 0 (control), 16, or 24‰ for 24h and then subjected to a critical swimming speed test (Ucrit) to quantify swimming ability. Following the test, the fish were weighed and blood samples were drawn to be analyzed for plasma ion and cortisol levels. While ion levels and weight loss were significantly higher in salt exposed fish, there were no significant differences in critical swimming speed or cortisol concentrations. This is in contrast to what has been observed in salmonids and Adriatic sturgeon. This suggests the hydromineral imbalance caused by moderate salt exposure is not sufficient to affect the swimming performance of shortnose sturgeon. Shortnose sturgeon are not thought to enter the saline stretches of the SJR until roughly 8years of age, yet this research shows that much younger juveniles withstand moderate salinity for short periods, with little whole-animal ramifications.

Hydrolysis of Extracellular Pyrophosphate increases in post-hemodialysis plasma

Vascular calcification (VC) is associated with significant morbidity and mortality of dialysis patients. Previous studies showed an association between loss of plasma pyrophosphate and VC. Moreover, loss of pyrophosphate occurs during dialysis in this population, suggesting that therapeutic approaches that prevent reduction of plasma pyrophosphate levels during dialysis could improve the quality of life of dialysis patients. This study found that pyrophosphate hydrolysis was 51% higher in post- than pre-dialysis plasma. Dialysis sessions modified the kinetic behavior of alkaline phosphatase, increasing its Vmax and reducing its Km, probably due to the elimination of uremic toxins during dialysis. At least 75% of alkaline phosphatase activity in human plasma was found to depend on a levamisole-sensitive enzyme probably corresponding to tissue non-specific alkaline phosphatase (TNAP). Dialysis increased total plasma protein concentration by 14% and reduced TNAP enzyme by 20%, resulting in an underestimation of pyrophosphate hydrolysis in post-dialysis plasma. Levamisole inhibited TNAP activity (IC50, 7.2 µmol/L), reducing pyrophosphate hydrolysis in plasma and increasing plasma pyrophosphate availability. Alkaline phosphatase is also found in many tissues and cells types; therefore, our results in plasma may be indicative of changes in phosphatase activity in other locations that collectively could contribute significantly to pyrophosphate hydrolysis in vivo. In conclusion, these findings demonstrate that dialysis increases pyrophosphate hydrolysis, which, taken together with previously reported increases in alkalization and calcium ion levels in post-dialysis plasma, causes VC and could be prevented by adding calcification inhibitors during dialysis.

Salinity acclimation and advanced parr–smolt transformation in growth-hormone transgenic coho salmon (Oncorhynchus kisutch)

Growth hormone (GH) is involved in the parr–smolt transformation of salmonid fishes and is known to improve salinity tolerance. This study compared the capacity for seawater acclimation of GH transgenic coho salmon (Oncorhynchus kisutch (Walbaum, 1792)) to that of wild-type fish, allowing examination of responses to sustained (chronic) exposure to elevated GH. GH transgenic fish (GH TG) smolted 1 year in advance of wild-type salmon and showed a greater capacity to hypo-osmoregulate in seawater. As GH TG fish were much larger than the wild-type fish, a second experiment was conducted with three size-matched groups of coho salmon (a 1+-year-old wild-type group, a 1+-year-old ration-restricted GH TG group, and a 0+-year-old fully fed GH TG group). When size-matched, the effect of GH transgenesis was not as dramatic, but the feed-rationed TG1+ group exhibited smaller deviations in plasma ion and osmolality levels following seawater exposure than did the other groups, suggesting a somewhat improved hypo-osmoregulatory ability. These results support a role for GH in the development of seawater tolerance by salmonid fishes independent of fish size.

The Structure and Nephroprotective Activity of Oligo-Porphyran on Glycerol-Induced Acute Renal Failure in Rats.

Porphyran is a sulfate galactan in the cell wall of Porphyra. Its acid hydrolysis product, oligo-porphyran (OP), was prepared and the structure studied by electrospray ionization time-of-flight mass spectrometry (ESI-TOF-MS). This oligosaccharide was mainly composed of monosulfate-oligo-galactan, disufate-oligo-galactan, trisulfate-oligo-galactan, trisulfate oligo-methyl-galactan, and 3,6-anhydrogalactose with the degree of polymerization ranging from 1 to 8. The effects of OP were investigated in the glycerol-induced acute renal failure (ARF) model. Compared with the normal group, rats from the glycerol-induced group exhibited collecting duct and medullary ascending limb dilation and casts. The OP-treated group exerted a protective effect against glycerol-induced changes. The results showed that the administration of OP markedly decreased mortality in female ARF rats. For male ARF rats, all of which survived, OP significantly decreased the blood urea nitrogen and serum creatinine levels. Ion levels in plasma and urine were significantly changed in ARF rats, whereas OP treatment almost recovered ion levels back to normal. This study showed a noticeable renal morphologic and functional protection by OP in glycerol-induced ARF rats.

Tolerance of piava juveniles to different ammonia concentrations

The objective of this study was to evaluate the tolerance of piavas (Leporinus obtusidens) to ammonia (NH3) by measuring its effects on plasma ion levels and tissue metabolic parameters. Piava juveniles (25-30 g) were exposed to five concentrations of NH3 (mg L-1): 0.003 (control), 0.1, 0.4, 0.7, and 1.4; after 96 hours, plasma levels of Na+, K+, Cl- and NH4+ ions and metabolic and enzyme activity in tissues (liver, kidneys, gills and muscle) were measured. The lethal concentration (LC50; 96h) of NH3 was 0.27 mg L-1. As NH3 increased, Na+ and NH4+ in the plasma increased and K+ decreased. In addition, Na+/K+- ATPase activity concomitantly increased in the gills and decreased in the kidneys. Glucose, glycogen, and protein levels decreased, while lactate and ammonia increased in the tissues of piava juveniles that were treated with higher concentrations of ammonia. The observed lethal toxicity could be due to a gradual depletion of plasma ion levels and a reduction of metabolic and Na+/K+-ATPase activity in tissues. Both can lead to dysfunction in ionoregulatory and physiological systems. This finding has implications for the management of fish culture of piavas.

Differential modulation of ammonia excretion, Rhesus glycoproteins and ion-regulation in common carp (Cyprinus carpio) following individual and combined exposure to waterborne copper and ammonia

The main objective of this study was to understand the mode of interaction between waterborne copper (Cu) and high environmental ammonia (HEA) exposure on freshwater fish, and how they influence the toxicity of each other when present together. For this purpose, individual and combined effects of Cu and HEA were examined on selected physiological and ion-regulatory processes and changes at transcript level in the common carp (Cyprinus carpio). Juvenile carp were exposed to 2.6μM Cu (25% of the 96h LC50value) and to 0.65mM ammonia (25% of the 96h LC50value) singly and as a mixture for 12h, 24h, 48h, 84h and 180h. Responses such as ammonia (Jamm) and urea (Jurea) excretion rate, plasma ammonia and urea, plasma ions (Na+, Cl− and K+), muscle water content (MWC) as well as branchial Na+/K+-ATPase (NKA) and H+-ATPase activity, and branchial mRNA expression of NKA, H+-ATPase, Na+/H+ exchanger (NHE-3) and Rhesus (Rh) glycoproteins were investigated under experimental conditions. Results show that Jamm was inhibited during Cu exposure, while HEA exposed fish were able to increase excretion efficiently. In the combined exposure, Jamm remained at the control levels indicating that Cu and HEA abolished each other’s effect. Expression of Rhcg (Rhcg-a and Rhcg-b) mRNA was upregulated during HEA, thereby facilitated ammonia efflux out of gills. On the contrary, Rhcg-a transcript level declined following Cu exposure which might account for Cu induced Jamm inhibition. Likewise, Rhcg-a was also down-regulated in Cu-HEA co-exposed fish whilst a temporary increment was noted for Rhch-b. Fish exposed to HEA displayed pronounced up-regulation in NKA expression and activity and stable plasma ion levels. In both the Cu exposure alone and combined Cu-HEA exposure, ion-osmo homeostasis was adversely affected, exemplified by the significant reduction in plasma [Na+] and [Cl−], and elevated plasma [K+], along with an elevation in MWC. These changes were accompanied by a decline in NKA activity. Gill H+-ATPase mRNA levels and activities were not affected by either Cu or HEA or both. Likewise, NHE-3 expression remained unaltered but tended to be numerically higher during HEA exposure. Overall, these data suggest that at equitoxic concentrations (25% of 96h LC50), the individual effect of Cu is more harmful while HEA induces quicker adaptive responses. Our findings also denote a competitive mode of interaction, exemplified by the inhibition of HEA -mediated adaptive responses in the presence of Cu.

Salinity effects on plasma ion levels, cortisol, and osmolality in Chinook salmon following lethal sampling

Studies on hydromineral balance in fishes frequently employ measurements of electrolytes following euthanasia. We tested the effects of fresh- or salt-water euthanasia baths of tricaine mesylate (MS-222) on plasma magnesium (Mg2+) and sodium (Na+) ions, cortisol and osmolality in fish exposed to saltwater challenges, and the ion and steroid hormone fluctuations over time following euthanasia in juvenile spring Chinook salmon (Oncorhynchus tshawytscha). Salinity of the euthanasia bath affected plasma Mg2+ and Na+ concentrations as well as osmolality, with higher concentrations in fish euthanized in saltwater. Time spent in the bath positively affected plasma Mg2+ and osmolality, negatively affected cortisol, and had no effect on Na+ concentrations. The difference of temporal trends in plasma Mg2+ and Na+ suggests that Mg2+ may be more sensitive to physiological changes and responds more rapidly than Na+. When electrolytes and cortisol are measured as endpoints after euthanasia, care needs to be taken relative to time after death and the salinity of the euthanasia bath.

Rab-GAP TBC1D4 (AS160) is dispensable for the renal control of sodium and water homeostasis but regulates GLUT4 in mouse kidney.

The Rab GTPase-activating protein TBC1D4 (AS160) controls trafficking of the glucose transporter GLUT4 in adipocytes and skeletal muscle cells. TBC1D4 is also highly abundant in the renal distal tubule, although its role in this tubule is so far unknown. In vitro studies suggest that it is involved in the regulation of renal transporters and channels such as the epithelial sodium channel (ENaC), aquaporin-2 (AQP2), and the Na+-K+-ATPase. To assess the physiological role of TBC1D4 in the kidney, wild-type (TBC1D4+/+) and TBC1D4-deficient (TBC1D4-/-) mice were studied. Unexpectedly, neither under standard nor under challenging conditions (low Na+/high K+, water restriction) did TBC1D4-/- mice show any difference in urinary Na+ and K+ excretion, urine osmolarity, plasma ion and aldosterone levels, and blood pressure compared with TBC1D4+/+ mice. Also, immunoblotting did not reveal any change in the abundance of major renal sodium- and water-transporting proteins [Na-K-2Cl cotransporter (NKCC2) NKCC2, NaCl cotransporter (NCC), ENaC, AQP2, and the Na+-K+-ATPase]. However, the abundance of GLUT4, which colocalizes with TBC1D4 along the distal nephron of TBC1D4+/+ mice, was lower in whole kidney lysates of TBC1D4-/- mice than in TBC1D4+/+ mice. Likewise, primary thick ascending limb (TAL) cells isolated from TBC1D4-/- mice showed an increased basal glucose uptake and an abrogated insulin response compared with TAL cells from TBC1D4+/+ mice. Thus, TBC1D4 is dispensable for the regulation of renal Na+ and water transport, but may play a role for GLUT4-mediated basolateral glucose uptake in distal tubules. The latter may contribute to the known anaerobic glycolytic capacity of distal tubules during renal ischemia.

Possible Effects of Metallosis on Spermatozoal Apoptotic Genes Expression in Individuals with Intramedullary Nailing Prosthesis

Seminal quality could be affected by metallosis caused by intramedullary nailing (IMN). Our objectives were to estimate metal ion levels in the seminal plasma of subjects with IMN, to determine their effects on semen parameters and on spermatozoal apoptotic gene expression, and to determine whether these expressed genes could be used as candidate biomarkers of seminal deterioration in individuals with IMN or not. Semen samples were collected from 60 subjects with IMN and 30 age-matched healthy controls. Seminal plasma contents of cobalt (Co), chromium (Cr), and molybdenum (Mo) were assayed. Spermatozoal Bcl-2 and Bax gene expressions were determined. Studied semen parameters were significantly lower in subjects with IMN for ≥5years in relation to controls while the concentrations of Co, Cr, and Mo in the seminal plasma samples were significantly higher. There were significantly lower spermatozoal Bcl-2 expression, higher Bax expression, and lower Bcl-2/Bax ratio in subjects with IMN for ≥5years than in controls. In subjects with IMN for ≥5years, receiver operating characteristic (ROC) curve analysis of studied gene expressions and Bcl-2/Bax ratio were done showing priority of the ratio with 86.7% sensitivity, 100% specificity, 100% positive predictive value, and 93.8% negative predictive value at cutoff values ≤0.777. Co, Cr, and Mo metals are found at high concentrations in the seminal plasma of individuals with IMN leading to increased spermatozoal apoptotic activity. Spermatozoal Bcl-2/Bax ratio could be used as a candidate biomarker of reproductive disorders in individuals with intramedullary nailing.

Humic acid and moderate hypoxia alter oxidative and physiological parameters in different tissues of silver catfish (Rhamdia quelen)

Aquatic animals are naturally exposed simultaneously to environments with different concentrations of humic acid (HA) and seasonal or daily variations of dissolved oxygen (DO) levels. This study investigated the effects of simultaneous exposure to different HA and DO levels on plasma ion levels and some hematological and oxidative parameters in different tissues of silver catfish (Rhamdia quelen). The fish were exposed to 0, 2.5 or 5mgL(-1) HA for 120h. After this period, each group was divided into two groups: normoxia and hypoxia. Exposure to the different DO levels lasted 96h, totaling 216h of experimentation. At the end of the experimental period, blood sampling was performed, and the fish were euthanized prior to the excision of the gills and the brain to evaluate hematological and oxidative parameters. To verify the antioxidant capacity of HA, total phenolic compounds were measured. In general, all tissues of silver catfish exposed simultaneously to hypoxia and different HA concentrations showed a reduction in lipid peroxidation levels, as well as a modulation of the antioxidant system. These effects occurred in an HA concentration-dependent manner. Thus, HA is beneficial to silver catfish exposed to hypoxia. These beneficial effects can be attributed, most likely, to the action of the different HA constituents and functional groups, including phenolic compounds, which have antioxidant properties.

The effect of ionization on the populations of excited levels of C IV and C V in tokamak edge plasmas

The main populating and depopulating mechanisms of the excited energy levels of ions in plasmas with densities <1023–1024 m−3 are electron collisional excitation from the ion's ground state and radiative decay, respectively, with the majority of the electron population being in the ground state of the ionization stage. Electron collisional ionization is predominately expected to take place from one ground state to that of the next higher ionization stage. However, the question arises as to whether, in some cases, ionization can also affect the excited level populations. This would apply particularly to those cases involving transient events such as impurity influxes in a laboratory plasma. An analysis of the importance of ionization in populating the excited levels of ions in plasmas typical of those found in the edge of tokamaks is undertaken for the C IV and C V ionization stages. The emphasis is on those energy levels giving rise to transitions of most use for diagnostic purposes (n ≤ 5). Carbon is chosen since it is an important contaminant of JET plasmas; it was the dominant low Z impurity before the installation of the ITER-like wall and is still present in the plasma after its installation. Direct electron collisional ionization both from and to excited levels is considered. Distorted-wave flexible atomic code calculations are performed to generate the required ionization cross sections, due to a lack of atomic data in the literature. Employing these data, ionization from excited level populations is not found to be significant in comparison with radiative decay. However, for some energy levels, ionization terminating in the excited level has an effect in the steady-state of the order of the measurement errors (±10%). During transient events, ionization to excited levels will be of more importance and must be taken into account in the calculation of excited level populations. More accurate atomic data, including possible resonance contributions to the cross sections, would tend to increase further the importance of these effects.