The acute kidney injury to chronic kidney disease transition in a mouse model of acute cardiorenal syndrome emphasizes the role of inflammation.

Acute cardiorenal syndrome is a common complication of acute cardiovascular disease. Studies of acute kidney injury (AKI) to chronic kidney disease (CKD) transition, including patients suffering acute cardiovascular disease, report high rates of CKD development. Therefore, acute cardiorenal syndrome associates with CKD, but no study has established causation. To define this we used a murine cardiac arrest (CA) and cardiopulmonary resuscitation (CPR) model or sham procedure on male mice. CA was induced with potassium chloride while CPR consisted of chest compressions and epinephrine eight minutes later. Two weeks after AKI was induced by CA/CPR, the measured glomerular filtration rate (GFR) was not different from sham. However, after seven weeks the mice developed CKD, recapitulating clinical observations. One day, and one, two, and seven weeks after CA/CPR, the GFR was measured, and renal tissue sections were evaluated for various indices of injury and inflammation. One day after CA/CPR, acute cardiorenal syndrome was indicated by a significant reduction of the mean GFR (649 in sham, vs. 25 µL/min/100g in CA/CPR animals), KIM-1 positive tubules, and acute tubular necrosis. Renal inflammation developed, with F4/80 positive and CD3-positive cells infiltrating the kidney one day and one week after CA/CPR, respectively. Although there was functional recovery with normalization of GFR two weeks after CA/CPR, deposition of tubulointerstitial matrix proteins α-smooth muscle actin and fibrillin-1 progressed, along with a significantly reduced mean GFR (623 in sham vs. 409 µL/min/100g in CA/CPR animals), proteinuria, increased tissue transforming growth factor-ß, and fibrosis establishing the development of CKD seven weeks after CA/CPR. Thus, murine CA/CPR, a model of acute cardiorenal syndrome, causes an AKI-CKD transition likely due to prolonged renal inflammation.

Pheretima aspergillum extract attenuates high-KCl-induced mitochondrial injury and pro-fibrotic events in cardiomyoblast cells.

Hyperkalemia is often associated with cardiac dysfunction. In this study an earthworm extract (dilong) was prepared from dried Pheretima aspergillum powder and its effect against high-KCl challenge was determined in H9c2 cardiomyoblast cells. H9c2 cells pre-treated with dilong (31.25, 62.5, 125, and 250 mg/mL) for 24 hours, where challenged with different doses of KCl treatment for 3 hours to determine the protective mechanisms of dilong against cardiac fibrosis. High-KCl administration induced mitochondrial injury and elevated the levels of pro-apoptotic proteins. The mediators of fibrosis such as ERK, uPA, SP1, and CTGF were also found to be upregulated in high-KCl condition. However, dilong treatment enhanced IGF1R/PI3k/Akt activation which is associated with cell survival. In addition, dilong also reversed high-KCl induced cardiac fibrosis related events in H9c2 cells and displayed a strong cardio-protective effect. Therefore, dilong is a potential agent to overcome cardiac events associated with high-KCl toxicity.

How to avoid severe coronary vasoconstriction in potassium induced cardioplegia.

OBJECTIVES: The aim of this study was to investigate how high K+ concentrations can be safely used in cardioplegic solutions without causing severe coronary artery vasocontraction. DESIGN: Twenty-four 50 kg pigs were used. The distal part of the left anterior descending coronary artery was cut into ring segments and transferred into organ baths with Krebs solution bubbled with 95% O2 and 5% CO2. K+ concentrations between 16 and 127 mM were used to induce vasocontractions at 37, 22, 15, and 8 °C. Mg2+ (0-20 mM) were used to attenuate K+ induced vasocontractions. RESULTS: K+-Krebs solution 127 mM at 37 °C induced a strong, sustained vasocontraction defined as 100%. The contractions induced by 16, 23, 30 and 127 mM K+ were: 7.7, 38, 72 and 100% at 37 °C; 1.7, 7.4, 21 and 65% at 22 °C; 1, 6.6, 15 and 33% at 15 °C; 0.6, 2.1, 6 and 14% at 8 °C, respectively. Mg2+ reduced the K+-induced contraction at 37 °C in a concentration-dependent way and Mg2+ at 8 mM practically eliminated the risk for severe vasocontraction. CONCLUSIONS: Hypothermia (8 °C) abolishes coronary contraction induced by K+-cardioplegic solutions. In normothermic cardioplegia 8 mM Mg2+ prevents vasoconstriction.

Toxicity of Potassium Chloride Compared to Sodium Chloride for Zebra Mussel Decontamination.

The use of chemicals to decontaminate watercraft and/or equipment after exposure to zebra mussels Dreissena polymorpha is one method of decontamination that has been recommended by multiple government agencies in the United States. The ideal chemical to be used for decontamination would be inexpensive and easily obtained, would have no or limited effect on nontarget species, and would be relatively environmentally friendly. Two chemicals that have been tested are potassium chloride (KCl) and sodium chloride (NaCl). The toxicity of each chemical to both adult zebra mussels and veliger larvae was examined. Sodium chloride was less effective at causing mortality than KCl within the exposure periods tested. Adult mussels required a 4× longer exposure period to exhibit complete mortality when exposed to NaCl at 30,000 mg/L (24 h) compared to KCl (6 h). At 10,000 mg/L, NaCl took 8× longer (96 h) than KCl (12 h) to cause 100% mortality of adult mussels. Veligers that were exposed to KCl at 1,250 mg/L required a 12-h exposure to attain complete mortality, while those exposed to NaCl at 10,000 mg/L required an 18-h exposure to exhibit the same result. To determine whether KCl is more advantageous as a decontamination chemical, the cost and chemical availability must be researched.

Protostane-Type Triterpenoids from Alisma orientale.

Twenty-eight protostane triterpenoids, including a new degraded one (1), nine new ones (2 - 10), and two new natural ones (11 and 12), have been isolated from the dried rhizomes of Alisma orientale. Alisol R (1) was the first 20,21,22,23,24,25,26,27-octanorprotostane triterpenoid. The absolute configurations of 25-methoxyalisol F (2) and 16ß-hydroperoxyalisol B 23-acetate (3) were determined by X-ray diffraction analysis. In addition, alismaketone-B 23-acetate (28) showed potent vasorelaxant activity on endothelium-intact thoracic aorta rings precontracted with KCl.

Alcohol-triggered signs of migraine: An animal model.

We describe an animal model where characteristics of migraine can be triggered by alcohol administration. In rats chronically implanted with a cannula overlying the transverse sinus, we applied potassium chloride (KCl) (or saline) to the meninges to sensitize trigeminovascular afferents. We assessed effects of repeated KCl application on animal behavior using conditioned place avoidance paradigm. In KCl-treated rats we discovered that alcohol injections (0.2 mg/kg), but not saline, resulted in the development of extracephalic allodynia and signs of ongoing pain.

Sensitivity of Danio rerio (Teleostei, Cyprinidae) during two stages of development based on acute toxicity tests.

The sensitivity of Danio rerio to three chemicals was compared at two growth stages [larval (10 ± 2 after hatching) and post-larval (60 ± 4 days after hatching)] based on acute toxicity tests. Thirty-nine 48 h acute toxicity tests were performed with the substances CuSO4, NaCl and KCl. The 48 h LC50 values at the two growth stages were compared by independent samples t-tests. The results showed a clear decrease in sensitivity when post-larval organisms were used. Since acute toxicity test methods for D. rerio that recommend using post-larval stage fish do not represent the most sensitive stage of the test organism, our study suggests a revision of the methods to use larval fish.

Evaluation of Chronic Effects of Potassium Chloride and Nickel on Survival, Growth, and Reproduction of a Unionid Mussel (Lampsilis siliquoidea).

The ASTM International standard test method for freshwater mussels (E2455-13) recommends 4-week toxicity testing with juveniles to evaluate chronic effects on survival and growth. However, concerns remain that the method may not adequately address the sensitivity of mussels to longer term exposures (>4 weeks), particularly in relation to potential reproductive impairments. No standard method directly evaluates toxicant effects on mussel reproduction. The objectives of the present study were to (1) evaluate toxicity endpoints related to reproduction in fatmucket (Lampsilis siliquoidea) using two common reference toxicants, potassium chloride (KCl) and nickel (Ni); (2) evaluate the survival and growth of juvenile fatmucket in standard 4-week and longer term (12-week) KCl and Ni tests following a method refined from the standard method; and (3) compare the sensitivity of the reproductive endpoints with the endpoints obtained from the juvenile mussel tests. Reproductive toxicity tests were conducted by first exposing female fatmucket brooding mature larvae (glochidia) to five test concentrations of KCl and Ni for 6 weeks. Subsamples of the glochidia were then removed from the adults to determine three reproductive endpoints: (1) the viability of brooded glochidia; (2) the viability of free glochidia in a 24-h exposure to the same toxicant concentrations as their mother; and (3) the success of glochidia parasitism on host fish. Mean viability of brooded glochidia was significantly reduced in the high KCl concentration (26 mg K/L) relative to the control, with a 20% effect concentration (EC20) of 14 mg K/L, but there were no significant differences between the control and any Ni treatment (EC20 > 95 µg Ni/L). The EC20s for viability of free glochidia after the additional 24-h exposure and parasitism success were similar to the EC20s of brooded glochidia. The EC20s based on the most sensitive biomass endpoint in the 4-week juvenile tests were 15 mg K/L and 91 µg Ni/L, similar to or greater than the EC20s from the reproductive KCl and Ni tests, respectively. When exposure duration in the juvenile tests was extended from 4 to 12 weeks, the EC20s decreased by more than 50% in the KCl test but by only 8% in the Ni test. Overall, these results indicate that a standard 4-week test with juvenile mussels can prove effective for estimating effects in chronic exposures with different life stages although a longer term 12-week exposure with juvenile mussels may reveal higher sensitivity of mussels to some toxicants, such as KCl. Environ Toxicol Chem 2024;43:1097-1111. © 2024 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Insights into the toxicological properties of a low molecular weight fraction from Zoanthus sociatus (Cnidaria).

The phylum Cnidaria is an ancient group of venomous animals, specialized in the production and delivery of toxins. Many species belonging to the class Anthozoa have been studied and their venoms often contain a group of peptides, less than 10 kDa, that act upon ion channels. These peptides and their targets interact with high affinity producing neurotoxic and cardiotoxic effects, and even death, depending on the dose and the administration pathway. Zoanthiniaria is an order of the Subclass Hexacorallia, class Anthozoa, and unlike sea anemone (order Actiniaria), neither its diversity of toxins nor the in vivo effects of the venoms has been exhaustively explored. In this study we assessed some toxicological tests on mice with a low molecular weight fraction obtained by gel filtration in Sephadex G-50 from Zoanthus sociatus crude extract. The gel filtration chromatogram at 280 nm revealed two major peaks, the highest absorbance corresponding to the low molecular weight fraction. The toxicological effects seem to be mostly autonomic and cardiotoxic, causing death in a dose dependent manner with a LD50 of 792 µg/kg. Moreover, at a dose of 600 µg/kg the active fraction accelerated the KCl-induced lethality in mice.

Influence of Water Hardness on Chronic Toxicity of Potassium Chloride to a Unionid Mussel (Lampsilis siliquoidea).

Elevated concentrations of potassium (K) often occur in effluents from wastewater treatment plants, oil and gas production operations, mineral extraction processes, and other anthropogenic sources. Previous studies have demonstrated that freshwater mussels are highly sensitive to K in acute and chronic exposures, and that acute toxicity of K decreases with increasing water hardness. However, little is known about the influence of hardness on the chronic toxicity of K. The objective of our study was to evaluate the chronic toxicity of K (tested as KCl) to a commonly tested unionid mussel (fatmucket, Lampsilis siliquoidea) at five hardness levels (25, 50, 100, 200, and 300 mg/L as CaCO3 ) representing most surface waters in the United States. Chronic 28-day K toxicity tests were conducted with 3-week-old juvenile fatmucket in the five hardness waters using an ASTM International standard method. The maximum acceptable toxicant concentrations (geometric mean of the no-observed-effect concentration and the lowest-observed-effect concentration) increased from 15.1 to 69.3 mg K/L for survival and from 15.1 to 35.8 mg K/L for growth (length and dry wt) and biomass when water hardness was increased from 25 mg/L (soft) to 300 mg/L (very hard). These results provide evidence to support water hardness influence on chronic K toxicity to juvenile fatmucket. However, the chronic effect concentrations based on the more sensitive endpoint (growth or biomass) increased only 2.4-fold from the soft water to the very hard water, indicating that water hardness had a limited influence on the chronic toxicity of K to the mussels. These results can be used to establish chronic toxicity thresholds for K across a broad range of water hardness and to derive environmental guideline values for K to protect freshwater mussels and other organisms. Environ Toxicol Chem 2023;42:1085-1093. Published 2023. This article is a U.S. Government work and is in the public domain in the USA.

An effective method for terrestrial arthropod euthanasia.

As scientific understanding of invertebrate life increases, so does the concern for how to end that life in an effective way that minimises (potential) suffering and is also safe for those carrying out the procedure. There is increasing debate on the most appropriate euthanasia methods for invertebrates as their use in experimental research and zoological institutions grows. Their popularity as pet species has also led to an increase in the need for greater veterinary understanding. Through the use of a local injection of potassium chloride (KCl) initially developed for use in American lobsters, this paper describes a safe and effective method for euthanasia in terrestrial invertebrates. Initial work focused on empirically determining the dose for cockroaches, which was then extrapolated to other arthropod species. For this method of euthanasia, we propose the term 'targeted hyperkalosis' to describe death through terminal depolarisation of the thoracic ganglia as a result of high potassium concentration.

Activation of meningeal nociceptors by cortical spreading depression: implications for migraine with aura.

Attacks of migraine with aura represent a phenomenon in which abnormal neuronal activity in the cortex produces sensory disturbances (aura) some 20-40 min before the onset of headache. The purpose of this study was to determine whether cortical spreading depression (CSD)--an event believed to underlie visual aura--can give rise to activation of nociceptors that innervate the meninges--an event believed to set off migraine headache. CSD was induced in anesthetized male rats by stimulation of the visual cortex with electrical pulses, pin prick, or KCl; single-unit activity of meningeal nociceptors was monitored in vivo in the rat before and after CSD. Regardless of the method of cortical stimulation, induction of CSD was recorded in 64 trials. In 31 of those trials, CSD induced a twofold increase in meningeal nociceptor firing rate that persisted for 37.0 +/- 4.6 min in trials in which activity returned to baseline, or >68 min in trials in which activity remained heightened at the time recording was interrupted. In two-thirds of the trials, onset of long-lasting neuronal activation began approximately 14 min after the wave of CSD. The findings demonstrates for the first time that induction of CSD by focal stimulation of the rat visual cortex can lead to long-lasting activation of nociceptors that innervate the meninges. We suggest that migraine with aura is initiated by waves of CSD that lead up to delayed activation of the trigeminovascular pathway.

Chronic daily cortical spreading depressions suppress spreading depression susceptibility.

BACKGROUND: Migraine is a disabling chronic episodic disorder. Attack frequency progressively increases in some patients. Incremental cortical excitability has been implicated as a mechanism underlying progression. Cortical spreading depression (CSD) is the electrophysiological event underlying migraine aura, and a headache trigger. We hypothesized that CSD events during frequent migraine attacks condition the cortex to increase the susceptibility to further attacks. METHODS: A single daily CSD was induced for 1 or 2 weeks in mouse frontal cortex; contralateral hemisphere served as sham control. At the end of CSD conditioning, occipital CSD susceptibility was determined by measuring the frequency of CSDs evoked by topical KCl application. RESULTS: Sham hemispheres developed 8.4 ± 0.3 CSDs/hour, and did not significantly differ from naïve controls without prior cranial surgery (9.3 ± 0.4 CSDs/hour). After 2 but not 1 week of daily CSD conditioning, CSD frequency (4.9 ± 0.3 CSDs/hour) as well as the duration and propagation speed were reduced significantly in the conditioned hemispheres. Histopathological examination revealed marked reactive astrocytosis without neuronal injury throughout the conditioned cortex after 2 weeks, temporally associated with CSD susceptibility. CONCLUSIONS: These data do not support the hypothesis that frequent migraine attacks predispose the brain to further attacks by enhancing tissue susceptibility to CSD.

Exenatide enhances kaliuresis under conditions of hyperkalemia.

Experiments on Wistar rats showed that exenatide (0.015-0.5 nmol per 100 g body weight) somewhat increased renal excretion of potassium from 7±1 to 16±1 µmol/h/100 g body weight (p<0.05) in animals with normal serum concentration of glucose (4.6±0.4 mM) and potassium (4.3±0.1 mM). Exenatide dramatically enhanced excretion of potassium under conditions of hyperkalemia (11.4±0.4 mM) produced by intraperitoneal injection of 1.25% KCl solution (5 ml per 100 g body weight). During the first postinjection hour, potassium excretion increased 2-fold and attained 97±11 µmol/h/100 g body weight in comparison with potassium load alone (47±9 µmol/h/100 g body weight, p<0.05). The data attest to a possible role of peptide regulators in normalization of potassium balance via renal mechanisms.

Development of a Reduced-Volume Acute Lethality Toxicity Test for Hyalella azteca.

Effects-directed analysis (EDA) is used to identify the principal toxic components within a complex mixture using iterative steps of chemical fractionation guided by bioassay results. Bioassay selection can be limited in EDA because of the volume requirements for many standardized test methods, and therefore, a reduced-volume acute toxicity test that also provides whole-organism responses is beneficial. To address this need, a static, 7-d, water-only, reduced-volume method (50 mL, 10 organisms) was developed for Hyalella azteca that substantially decreases the volume requirements of standard-volume acute test exposures (200-500 mL of test solution, 15-20 organisms) while maintaining water quality and meeting control survival criteria. Standard- and reduced-volume methods were compared by conducting concurrent toxicity tests with 2 inorganic toxicants (KCl and CdCl2 ) and 2 organic mixtures of naphthenic acid fraction components (NAFCs) to evaluate test performance. There was no difference between methods when comparing the median lethal concentrations (LC50s) for KCl and both NAFC mixtures (p > 0.05). The LC50s for CdCl2 were statistically different (p = 0.0002); however, this was not considered biologically meaningful because the difference between LC50s was <2-fold. In conclusion, the reduced-volume H. azteca test method generated results comparable to standard-volume test methods and is suitable for use in situations where limited testing material is available, such as when conducting EDA. Environ Toxicol Chem 2020;39:2221-2227. © Her Majesty the Queen in Right of Canada 2020. Reproduced with the permission of the Minister of Environment and Climate Change Canada.

Subchronic exposure to nitrite, potassium and their combination on survival, growth, total haemocyte count and gill structure of juvenile blue swimmer crabs, Portunus pelagicus.

Among anthropogenic discharges, nitrite and potassium can simultaneously enter aquatic ecosystems at high loading rates which can threaten marine animals. The sensitivity of early juvenile blue swimmer crabs, Portunus pelagicus, to the subchronic exposure to nitrite, potassium and their combination was evaluated by measuring the survival, growth, total haemocyte count (THC) and gill histopathological changes. In all NaNO(2)-N and KNO(2)-N treatments the survival substantially decreased, many due to "molt death syndrome", along with significantly lower (p<0.01) specific growth rates (SGR). Although fewer deaths occurred in the KCl-K treatments, the SGR at the higher concentrations were significantly less (p<0.05) than the control. The gill histopathological changes following elevated NaNO(2)-N, KNO(2)-N and KCl-K exposure showed drastic but similar damage. In spite of a long term healing response, indicated by a significant hemolymph THC increase (p<0.01) and several gill lamellae modifications, early P. pelagicus juveniles are highly sensitive to elevated NO(2)-N levels.

Neuroprotective effects of ginseng saponins against L-type Ca2+ channel-mediated cell death in rat cortical neurons.

In the present study, we have examined any possible involvement of L-type Ca(2+) channels in ginseng-mediated neuroprotective actions. Exposure to a 50mM KCl (high-K) produced neuronal cell death, which was blocked by a selective L-type Ca(2+) channel blocker in cultured cortical neurons. When cultured cells were co-treated with ginseng total saponin (GTS) and high-K, GTS reduced high-K-induced neuronal death. Using Ca(2+) imaging techniques, we found that GTS inhibited high-K-mediated acute and long-term [Ca(2+)](i) changes. These GTS-mediated [Ca(2+)](i) changes were diminished by nifedipine. Furthermore, GTS-mediated effects were also diminished by a saturating concentration of Bay K (10muM). After confirming the protective effect of GTS using a TUNEL assay, we found that ginsenosides Rf and Rg(3) are active components in ginseng-mediated neuroprotection. These results suggest that inhibition of L-type Ca(2+) channels by ginseng could be one of the mechanisms for ginseng-mediated neuroprotection in cultured rat cortical neurons.

Dual neurotoxic and neuroprotective role of metabotropic glutamate receptor 1 in conditions of trophic deprivation - possible role as a dependence receptor.

Group I metabotropic glutamate receptors have been often implicated in various models of neuronal toxicity, however, the role played by the individual receptors and their putative mechanisms of action contributing to neurotoxicity or neuroprotection remain unclear. Here, using primary cultures of rat cerebellar granule cells and mouse cortical neurons, we show that conditions of trophic deprivation increased mGlu1 expression which correlated with the developing cell death. The inhibition of mGlu1 expression by specific siRNA attenuated toxicity, while adenovirus-mediated overexpression of mGlu1 resulted in increased cell death, indicating a causal relationship between the level of receptor expression and neuronal survival. In pharmacological experiments selective mGlu1 antagonists failed to protect from mGlu1-induced cell death, instead, neuronal survival was promoted by glutamate acting at mGlu1 receptors. Such properties are characteristics of a novel heterogeneous family of dependence receptors which control neuronal apoptosis. Our findings indicate that increased expression of mGlu1 in neurons creates a state of cellular dependence on the presence of its endogenous agonist glutamate. We propose a new role and a new mechanism for mGlu1 action. This receptor may play a crucial role in determining the fate of individual neurons during the development of the nervous system.

Inherent toxicity of organ preservation solutions to cultured hepatocytes.

Organ preservation solutions have been designed to protect grafts against the injury inflicted by cold ischemia. However, toxicity of University of Wisconsin (UW) solution during rewarming has been reported. Therefore, we here assessed the toxicity of UW, histidine-tryptophan-ketoglutarate (HTK), Euro-Collins, histidine-lactobionate (HL), sodium-lactobionate-sucrose and Celsior solutions in cultured hepatocytes under hypothermic (4 degrees C), intermediate (21 degrees C) and physiological (37 degrees C) conditions. Marked toxicity of UW, HTK, HL and Euro-Collins solutions was observed at both 37 and 21 degrees C. With the exception of UW solution, these solutions also increased cell injury during cold incubation (LDH release after 18 h at 4 degrees C: HTK 76+/-2%, Euro-Collins 78+/-17%, HL 81+/-15%; control: Krebs-Henseleit buffer 20+/-6%). Testing of individual components using modified Krebs-Henseleit buffers suggested that histidine and phosphate are responsible for (part of) this toxicity. These potential toxicities should be taken into account in the development of future preservation solutions.

Differences in ventricular vulnerability, serum enzyme activities, and electrolyte levels between male and female broilers.

Sudden death syndrome (SDS) is one of the most serious diseases of fast-growing broilers. The incidence of SDS may result from a decrease in ventricular function. The purpose of this study was to explore the mechanism of sexual difference in the sensitivity of broilers to SDS by measuring their ventricular vulnerability, serum enzyme activities, and serum electrolyte levels. Results were as follows. 1) Ventricular fibrillation thresholds induced by injection of KCl and by electrical stimulus of male broilers were both significantly lower than those of female broilers (P < 0.05), suggesting that the ventricular vulnerability of male broilers was higher than that of female broilers. 2) Serum lactate dehydrogenase and creatine kinase activities of male broilers were significantly higher than those of female broilers (P < 0.01), but there was not a significant difference in serum aspartate aminotransferase activity between male and female broilers. 3) No significant difference was observed in serum electrolyte levels of potassium, sodium, and chloride between males and females. From these results, we concluded that there is a significant difference between males and females in their ventricular vulnerability and serum enzyme activities, which may result in a higher sensitivity of male broilers to injury of the myocardium by stress and may further result in a sexual difference in sensitivity to SDS.