Acute Experiments Research Articles

Fetal drug exposure after maternally administered CFTR modulators Elexacaftor/Tezacaftor/Ivacaftor in a rat model

BackgroundThe potential effects of the very effective cystic fibrosis triple combination drug, Elexacaftor/Tezacaftor/Ivacaftor (ETI) in pregnancy on prenatal development of offspring remain largely unknown. Research questionWe aimed to investigate the fetal tissue distribution pattern of maternally administered ETI by placental transfer in the rat fetuses. Study design and methodsSprague Dawley pregnant rats were administered ETI (6.7 mg/kg/d elexacaftor + 3.5 mg/kg/d tezacaftor + 25 mg/kg/d ivacaftor) traced with [3 H]-ivacaftor in single dose acute experiments (intraperitoneal injection) or treated orally with ETI (the same dose) for 7 days in sub-chronic experiments. Fetal tissue samples were collected at embryonic day (E) 19 and analyzed using liquid scintillation counting for acute experiments or liquid chromatography-mass spectrometry for sub-chronic experiments. ResultsOn day E19, after acute exposure, the entry of ivacaftor into fetal brain (brain/plasma concentration ratios <50%) was significantly lower than to other tissues (>100%). However, after sub-chronic exposure, the entry of all 3 components into the developing brain was comparably extensive as into other tissues (tissue/plasma ratios, 260 – 1000%). Each component of ETI accumulated in different fetal tissues to approximately equal extent. Inter-litter differences on fetal drug distribution were found in cortex for ivacaftor, muscle for tezacaftor and cortex and mid/hindbrain for elexacaftor. Fetal plasma concentrations of ETI (ng/mL) were variable between litters. The entry of ivacaftor and tezacaftor into adult brain appeared to be restricted (<100%). InterpretationFetal rats are exposed to maternally ingested ETI after sub-chronic exposure, potentially impacting fetal development. The brain entry data highlights the need for attention be paid to any long-term potential effects ETI exposure could have on normal brain development.

Personalized inference for neurostimulation with meta-learning: a case study of vagus nerve stimulation.

Objective. Neurostimulation is emerging as treatment for several diseases of the brain and peripheral organs. Due to variability arising from placement of stimulation devices, underlying neuroanatomy and physiological responses to stimulation, it is essential that neurostimulation protocols are personalized to maximize efficacy and safety. Building such personalized protocols would benefit from accumulated information in increasingly large datasets of other individuals' responses.Approach. To address that need, we propose a meta-learning family of algorithms to conduct few-shot optimization of key fitting parameters of physiological and neural responses in new individuals. While our method is agnostic to neurostimulation setting, here we demonstrate its effectiveness on the problem of physiological modeling of fiber recruitment during vagus nerve stimulation (VNS). Using data from acute VNS experiments, the mapping between amplitudes of stimulus-evoked compound action potentials (eCAPs) and physiological responses, such as heart rate and breathing interval modulation, is inferred.Main results. Using additional synthetic data sets to complement experimental results, we demonstrate that our meta-learning framework is capable of directly modeling the physiology-eCAP relationship for individual subjects with much fewer individually queried data points than standard methods.Significance. Our meta-learning framework is general and can be adapted to many input-response neurostimulation mapping problems. Moreover, this method leverages information from growing data sets of past patients, as a treatment is deployed. It can also be combined with several model types, including regression, Gaussian processes with Bayesian optimization, and beyond.

Hypertensive rats show increased renal excretion and decreased tissue concentrations of glycine betaine, a protective osmolyte with diuretic properties.

Hypertension leads to water-electrolyte disturbances and end-organ damage. Betaine is an osmolyte protecting cells against electrolyte imbalance and osmotic stress, particularly in the kidneys. This study aimed to evaluate tissue levels and hemodynamic and renal effects of betaine in normotensive and hypertensive rats. Betaine levels were assessed using high-performance liquid chromatography-mass spectrometry (HPLC-MS) in normotensive rats (Wistar-Kyoto, WKYs) and Spontaneously Hypertensive rats (SHRs), a model of genetic hypertension. Acute effects of IV betaine on blood pressure, heart rate, and minute diuresis were evaluated. Gene and protein expression of chosen kidney betaine transporters (SLC6a12 and SLC6a20) were assessed using real-time PCR and Western blot. Compared to normotensive rats, SHRs showed significantly lower concentration of betaine in blood serum, the lungs, liver, and renal medulla. These changes were associated with higher urinary excretion of betaine in SHRs (0.20 ± 0.04 vs. 0.09 ± 0.02 mg/ 24h/ 100g b.w., p = 0.036). In acute experiments, betaine increased diuresis without significantly affecting arterial blood pressure. The diuretic response was greater in SHRs than in WKYs. There were no significant differences in renal expression of betaine transporters between WKYs and SHRs. Increased renal excretion of betaine contributes to decreased concentration of the protective osmolyte in tissues of hypertensive rats. These findings pave the way for studies evaluating a causal relation between depleted betaine and hypertensive organ damage, including kidney injury.

ЭКСПЕРИМЕНТАЛЬНЫЕ ВОЗМОЖНОСТИ ИССЛЕДОВАНИЯ ТОНУСА ПРИ НАРУШЕНИИ СОКРАТИТЕЛЬНОЙ ФУНКЦИИ ГЛАДКИХ МИОЦИТОВ

A method has been proposed for assessing the functional state of the uterus of experimental pregnant animals, including simultaneous recording of signals from the electrical and mechanical activity of the uterus using a kymograph and an electromyocytograph. The method allows you to record the frequency and duration of uterine contractions and record a mechanogram, and timely visualize uterine hypotension during the administration of various doses of uterotonics. This will allow us to establish the degree of development of myometrial hypotension under the influence of excessive doses of exogenous uterotonics. The combined research method will allow assessing the functional state of the uterus of a pregnant rat against the background of induced uterine contractions in an acute experiment.

КОРРЕКЦИЯ КИСЛОРОДНОГО ГОЛОДАНИЯ В ЭКСПЕРИМЕНТАХ НА ЖИВОТНЫХ ПРИ МОДЕЛИРОВАНИИ ЭФФЕКТОВ, ВЫЗЫВАЕМЫХ ОСТРОЙ ДЫХАТЕЛЬНОЙ НЕДОСТАТОЧНОСТЬЮ, С ПОМОЩЬЮ МНОГОКОМПОНЕНТНОЙ ГАЗОВОЙ СМЕСИ НА ОСНОВЕ КОМБИНАЦИИ ИНЕРТНЫХ ГАЗОВ

Acute respiratory insufficiency (ARI) accompanied with hypoxemia and hypercapnia requires oxygen therapy. The helium-oxygen therapy secures from the undesirable effects of oxygen therapy. Earlier we showed that introduction of argon in a breathing mixture (BM) reduces the effect of hypoxia. Purpose of the work is to test the potentiality of BM containing a combination of inert gases to counteract oxygen starvation in acute ARI-provoking experiments with animals. Fourteen Wistar rats with a body mass of 276 ± 14.6 grams were divided into two groups. An integral noninvasive technology was used for real-time assessment of oxygen transport and consumption in each animal. Both the experiment and control animals breathed a mixture with a low O2 and high CO2 concentration. The control animals received a routine therapy by a hyperoxic mixture containing nitrogen and carbon dioxide. The experimental animals were treated with a hyperoxic mixture containing helium and argon, and carbon dioxide to induce hypercapnia. The results demonstrated reliably that the hyperoxic heluim-argon mixture reduces burden on the external respiration and favors oxygen utilization in tissues; however, this happens at the expense of a greater stress to the cardiovascular system. In contrast, the hyperoxic mixture used in the control stressed the external respiration rather than the cardiac muscle.

Degradation of Angelica sinensis polysaccharide: Structures and protective activities against ethanol-induced acute liver injury

Angelica sinensis polysaccharide (ASP) possesses diverse bioactivities; however, its metabolic fate following oral administration remains poorly understood. To intuitively determine its intestinal digestion behavior after oral administration, ASP was labeled with fluorescein, and it was found to accumulate and be degraded in the cecum and colon. Therefore, we investigated the in vitro enzymatic degradation behavior and identified the products. The results showed that ASP could be degraded into fragments with molecular weights similar to those of the fragments observed in vivo. Structural characterization revealed that ASP is a highly branched acid heteropolysaccharide with AG type II domains, and its backbone is predominantly composed of 1,3-Galp, →3,6)-Galp-(1→6)-Galp-(1→, 1,4-Manp, 1,4-Rhap, 1,3-Glcp, 1,2,3,4-Galp, 1,3,4,6-Galp, 1,3,4-GalAp and 1,4-GlcAp, with branches of Araf, Glcp and Galp. In addition, the high molecular weight enzymatic degradation products (ASP H) maintained a backbone structure almost identical to that of ASP, but exhibited only partial branch changes. Then, the results of ethanol-induced acute liver injury experiments revealed that ASP and ASP H reduced the expression of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), and malondialdehyde (MDA) and increased the superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) levels, thereby relieving ethanol-induced acute liver injury.

Long-term normothermic autoperfusion of the cardiopulmonary complex ex vivo as a method of effective graft conditioning: an experimental study

Objective: To compare the effectiveness of 6-hour normothermic autoperfusion of a heart graft ex vivo with pharmaco-cold preservation using Bretschneider's solution (Custodiol, Dr Franz Köhler Chemie GmbH, Bensheim, Germany).Methods: Landrace pigs weighing 50 ± 5 kg and aged 4–5 months (n = 10) were selected as a model for a series of acute experiments. Cardiopulmonary conditioning using autoperfusion was conducted for 6 hours on the experimental group (n = 5). On the other hand, the control group underwent a 6-hour pharmaco-cold preservation with Bretschneider solution to recover the heart's pumping function. Graft preservation effectiveness was evaluated by measuring hemodynamic parameters, heartbeat, and myocardial ischemia marker concentrations.Results: After reperfusion and isolation of the working cardiopulmonary complex, cardiac output was 0.63 [0.37; 0.8] L/min and 0.37 [0.23; 0.37] L/min in the experimental and control groups, respectively (P &lt; .05). The levels of CPK-MB, LDH, troponin-I, and lactate in the coronary sinus blood was significantly higher in the control group.Conclusion: The study demonstrated significant benefits of normothermic autoperfusion in maintaining the morphofunctional status of the donor heart compared to pharmaco-cold preservation using Bretschneider's solution for 6 hours of ex vivo graft conditioning. Received 16 July 2023. Revised 8 September 2023. Accepted 11 September 2023. Funding: The study was carried out within the framework of project No. 23-25-10013 (agreement No. 23-25-10013 dated April 20, 2023 with the Russian Science Foundation, agreement No. р-52 dated April 3, 2023 with the Ministry of Science and Innovation Policy of the Novosibirsk Region). Conflict of interest: The authors declare no conflict of interest. Contribution of the authorsConception and study design: M.O. Zhulkov, D.A. Sirota, I.S. ZykovData collection and analysis: M.O. Zhulkov, A.R. Tarkova, I.S. Zykov, A.G. Makaev, A.V. Protopopov, M.N. Murtazaliev, F.Yu. Kosimov, N.A. Karmadonova, Ya.M. Smirnov, E.E. Kliver, A.M. Volkov, H.A. Agaeva, D.A. SirotaStatistical analysis: M.O. ZhulkovDrafting the article: M.O. ZhulkovCritical revision of the article: M.O. Zhulkov, D.A. Sirota, I.S. ZykovFinal approval of the version to be published: M.O. Zhulkov, A.R. Tarkova, I.S. Zykov, A.G. Makaev, A.V. Protopopov, M.N. Murtazaliev, F.Yu. Kosimov, N.A. Karmadonova, Ya.M. Smirnov, E.E. Kliver, A.M. Volkov, H.A. Agaeva, D.A. Sirota

Genome-Wide Association Analysis of Heat Tolerance in F2 Progeny from the Hybridization between Two Congeneric Oyster Species.

As the world's largest farmed marine animal, oysters have enormous economic and ecological value. However, mass summer mortality caused by high temperature poses a significant threat to the oyster industry. To investigate the molecular mechanisms underlying heat adaptation and improve the heat tolerance ability in the oyster, we conducted genome-wide association analysis (GWAS) analysis on the F2 generation derived from the hybridization of relatively heat-tolerant Crassostrea angulata ♀ and heat-sensitive Crassostrea gigas ♂, which are the dominant cultured species in southern and northern China, respectively. Acute heat stress experiment (semi-lethal temperature 42 °C) demonstrated that the F2 population showed differentiation in heat tolerance, leading to extremely differentiated individuals (approximately 20% of individuals die within the first four days with 10% survival after 14 days). Genome resequencing and GWAS of the two divergent groups had identified 18 significant SNPs associated with heat tolerance, with 26 candidate genes located near these SNPs. Eleven candidate genes that may associate with the thermal resistance were identified, which were classified into five categories: temperature sensor (Trpm2), transcriptional factor (Gata3), protein ubiquitination (Ube2h, Usp50, Uchl3), heat shock subfamily (Dnajc17, Dnaja1), and transporters (Slc16a9, Slc16a14, Slc16a9, Slc16a2). The expressional differentiation of the above genes between C. gigas and C. angulata under sublethal temperature (37 °C) further supports their crucial role in coping with high temperature. Our results will contribute to understanding the molecular mechanisms underlying heat tolerance, and provide genetic markers for heat-resistance breeding in the oyster industry.

Menthol dural application alters meningeal arteries tone and enhances excitability of trigeminocervical neurons in rats

Headaches, including migraines, can have a causal relationship to exposure to cold, and this relationship may be both positive and negative, as cold can both provoke and alleviate cephalgia. The role of thermoreceptors responsible for transduction of low temperatures belongs to the transient receptor potential cation channel subfamily melastatin member 8 (TRPM8). These channels mediate normal cooling sensation and have a role in both cold pain and cooling-mediated analgesia; they are seen as a potential target for principally new anti-migraine pharmaceuticals. Using a validated animal migraine models, we evaluated effects of menthol, the TRPM8–agonist, on trigeminovascular nociception.In acute experiments on male rats, effects of applied durally menthol solution in various concentrations on the neurogenic dural vasodilatation (NDV) and firing rate of dura-sensitive neurons of the trigeminocervical complex (TCC) were assessed.Application of menthol solution in concentrations of 5 % and 10 % was associated with NDV suppression, however amplitude reduction of the dilatation response caused not by the vascular dilatation degree decrease, but rather due to the significant increase of the meningeal arterioles’ basal tone. In electrophysiological experiments the 1 % and 30 % menthol solutions intensified TCC neuron responses to the dural electrical stimulation while not changing their background activity.Revealed in our study excitatory effects of menthol related to the vascular as well as neuronal branches of the trigeminovascular system indicate pro-cephalalgic effects of TRPM8-activation and suggest feasibility of further search for new anti-migraine substances among TRPM8-antagonists.

Regulation of cadmium in the soil according to its effect on plants

Introduction. There is discussed the influence of various concentrations of Cd in the soil taking into account their established limiting values on plants in the publication.&#x0D; Materials and methods. An express experiment was performed with Cd concentrations (0.5; 1.0; 2.5; 5.0 mg/kg) on Avena sativa L. and Sinapis alba L. using the phytotesting method. In a series of vegetation experiments, cultures of Lactuca sativa var. capitata and Hordeum vulgare L. were used. In this case, the Cd concentration increased to 15 mg/kg.&#x0D; Results. A pronounced level of toxicity (&gt;50%) is manifested in the concentration of cadmium above 5 mg/kg, as established in preliminary experiments &#x0D; on podzolic soil. The results of vegetation studies indicate a pronounced negative effect of cadmium at concentrations above 5 mg/kg, regardless of the forms of its intake (State standard sample (SSS) and Cd(NO3)2). The biomass of barley and lettuce plants) is reduced by 54 and 36%, respectively (Cd in the form of SSS) and by 35 and 44% (Cd in the form of Cd(NO3)2). An increase in the content of Cd in the soil at a level of ~8–13 mg/kg accompanied by a high level of its accumulation in plants from 18.5 to 33.0 µg/g.&#x0D; Limitations. When studying the effect of cadmium at concentrations of 0.5–15 mg/kg in soil on plants in acute and chronic experiments, the following crops were used: Avena sativa L. and Sinapis alba L., Hordeum vulgare L. and Lactuca sativa var. To restrict a chemical element in the soil-plant system, one should strive to extend the duration of the study and diversify the choice of crops.&#x0D; Conclusion. The tested doses of cadmium may have some negative effects and increase the risk to human health. Doses of cadmium corresponding to the approximate permissible concentrations of this element in the soil reduce the quality of plant material, confirmed by its high concentrations.

Acute and chronic effects of triclosan on the behavior, physiology, and multigenerational characteristics of the water flea Moina macrocopa

Triclosan, a chlorinated biphenyl ether is widely used in industrial products and cosmetics due to its antibiotic activity. Although relatively levels of triclosan have been detected in aquatic ecosystems, limited information is available regarding the acute and chronic impacts of triclosan on aquatic invertebrates, especially planktonic crustaceans. In this study, we analyzed the acute (24 h) and chronic (14 days exposure across three generations) effects of different concentrations of triclosan [1/10 of the no observed effect concentration (NOEC), the NOEC, and 1/10 of the LC50] calculated from the 24 h acute toxicity value, on the water flea Moina macrocopa. In the acute exposure experiment, the 1/10 LC50 value of triclosan significantly reduced survival, feeding rate, thoracic limb activity, heart activity, and acetylcholinesterase activity. In response to the 1/10 LC50 value, intracellular reactive oxygen species increased along with elevated levels of malondialdehyde and glutathione. Enzymatic activities of catalase, superoxide dismutase, glutathione peroxidase, and glutathione reductase were significantly increased by the 1/10 LC50 value, suggesting active protection of the antioxidant defense system against oxidative stress. Chronic exposure to the 1/10 NOEC and NOEC values revealed multigenerational adverse impacts of triclosan. The second generation was found to be the most sensitive to triclosan, as the NOEC value significantly reduced the survival rate, body length, and the number of neonates per brood, along with a delayed hatching period. Taken together, these results indicate that even sublethal levels of triclosan can have detrimental effects on the water flea population's maintenance through intergenerational toxicity.

Potential Antihypertensive Activity of the Aqueous Extract of Ammi visnaga and its Effect on ACE-2 in Rats.

This work aimed to investigate the antihypertensive activity of Ammi visnaga. The aqueous extract of Ammi visnaga has traditionally been used to treat hypertension in Morocco. The objective of this investigation was to evaluate the effect of Ammi visnaga aqueous extract (AVAE) on arterial blood pressure, systolic blood pressure (SBP), mean blood pressure (MBP), diastolic blood pressure (DBP), and heart rate (HR) in normotensive and hypertensive rats. In addition, the effect of the aqueous extract of Ammi visnaga on vasodilatation was assessed in isolated rat aortic rings with functional endothelium pre-contracted with epinephrine EP or KCl. AVAE was obtained, and its antihypertensive ability was pharmacologically investigated in L-NAME hypertensive and normotensive rats. The rats received oral AVAE at two selected doses of 70 and 140 mg/kg for six hours (acute experiment) and seven days (sub-chronic). Thereafter, systolic, diastolic, mean arterial blood pressure and heart rate were evaluated. Moreover, the vasorelaxant activity of AESA was performed in thoracic aortic ring rats. In addition, the mechanisms of action involved in the vasorelaxant effect were studied. AVAE lowered blood pressure only in L-Name-induced hypertensive rats. Furthermore, AVAE (0.375-1.375 mg/ml) showed a vasodilator effect in isolated aortic rats. In addition, not all of the medications used in our study were responsible for the signaling pathway. As a result, additional pharmaceuticals are required to confirm the mechanism of this signaling pathway. The aqueous extract of Ammi visnaga exerts an interesting antihypertensive activity, which could be mediated through its vasorelaxant activity. The study supports its use as a medicinal plant against hypertension in Morocco.

Acetaminophen-traces bioremediation with novel phenotypically and genotypically characterized 2 Streptomyces strains using chemo-informatics, in vivo, and in vitro experiments for cytotoxicity and biological activity

We isolated two novel bacterial strains, active against the environmental pollutant acetaminophen/Paracetamol®. Streptomyces chrestomyceticus (symbol RS2) and Flavofuscus (symbol M33) collected from El-Natrun Valley, Egypt—water, sediment, and sand samples, taxonomically characterized using a transmission electron microscope (TEM). Genotypic identification, based on 16S rRNA gene sequence analysis followed by BLAST alignment, were deposited on the NCBI as 2 novel strains https://www.ncbi.nlm.nih.gov/nuccore/OM665324 and https://www.ncbi.nlm.nih.gov/nuccore/OM665325. The phylogenetic tree was constructed. Acetaminophen secondary or intermediate product’s chemical structure was identified by GC/LC MS. Some selected acetaminophen secondary-product extracts and derived compounds were examined against a panel of test micro-organisms and fortunately showed a good anti-microbial effect. In silico chemo-informatics Swiss ADMET evaluation was used in the selected bio-degradation extracts for absorption (gastric), distribution (to CNS), metabolism (hepatic), excretion (renal), and finally not toxic, being non-mutagenic/teratogenic or genotoxic, virtually. Moreover, in vitro cytotoxic activity of these selected bio-degradation secondary products was examined against HepG2 and MCF7 cancer cell lines, where M33 and RS2 extract effects on acetaminophen/paracetamol bio-degradation products were safe, with higher IC50 on HepG2 and MCF7 than the acetaminophen/paracetamol IC50 of 108.5 μg/ml. Moreover, an in vivo oral acute single-dose toxicity experiment was conducted, to confirm these in vitro and in silico lower toxicity (better safety) than acetaminophen/paracetamol.

Effect of sulfide on the physiological energetics of Mercenaria mercenaria

Marine sulfide is mainly produced by the reduction of sulfate. Eutrophication and anoxic conditions caused by tidal fluctuations have accelerated the production of sulfide and widened its distribution. Sulfide has toxic effects on aquatic animals, and it has important effects on physiological activities such as feeding and respiration in buried bivalves. The aim of this study was to estimate the impact of sulfide on the energy budget of the intertidal bivalve Mercenaria mercenaria. Acute toxicity experiments were first conducted on small (SS), medium (MS), and large (LS) size clams, and then clearance rate (CR), absorption efficiency (AE), respiration rate (RR), and ammonia excretion rate (ER) were measured at 24, 48, 72, and 96 h using 0 (control group), safe concentration (low sulfide group), and 96 h half-lethal concentration (high sulfide group) as sulfide concentration gradients, and scope for growth (SFG) was calculated. The results indicated that sulfide reduced the CR, AE, RR, and ER of M. mercenaria, and these indicators were lower in the high sulfide group than in the low sulfide group. The sulfide concentration and exposure time had significant effects on CR, AE, RR, and SFG. The CR of the SS low sulfide group reached its maximum value at 48 h, and the CR of the LS low sulfide and high sulfide groups reached their maximum values at 96 h and 48 h, respectively. SFG experienced a significant decrease with the extension of time, and for both MS and LS, the SFG of sulfide-treated groups was negative at all times, with the exception of 24 h. These results suggest that M. mercenaria exhibits metabolic inhibition and metabolic compensation in response to sulfide exposure. This study provides important insights for comprehending the physiological response of buried bivalves in sulfide environments and provides a scientific fundation for implementing corresponding culture measures.

Toxicity study of compound granules of Hedyotis diffusa: Acute toxicity and long-term toxicity

Ethnopharmacological relevanceThe clinical efficacy of the hospital preparation compound granules of Hedyotis diffusa (CGHD), which is composed of Hedyotis diffusa Willd, Smilax china L., Solanum lyratum Thunb., has accumulated a good reputation over the past decades. However, because it is a hospital preparation, few researchers have paid attention to it, resulting in a lack of systematic basic research studies. Thus, it is not clear whether there are safety concerns that restrict its clinical application, and toxicological evaluation of CGHD is needed. Aim of the studyThe aim of this study was to evaluate the safety of CGHD by conducting acute toxicity and long-term toxicity experiments, with the objective of providing evidence for its clinical safety and a theoretical foundation for its clinical application. Materials and methodsKM mice were selected for the acute toxicity experiment and were administered water or CGHD-E 3 times within 24 h. The reactions of the animals to CGHD treatment were observed and recorded within 1 h after administration and then once a day for 14 consecutive days. SD rats were selected to conduct the long-term toxicity experiment. The drug-treated groups were administered different doses of CGHD-E, which were equivalent to 10 times, 20 times and 50 times the clinical dose in humans. The rats were administered the drug for 28 consecutive days. After 28 days, the animals were sacrificed, and routine blood tests, blood coagulation function analysis, liver and kidney function tests, and glycolipid metabolism related tests were conducted. The major organs of the rats were collected to calculate organ coefficients and perform hematoxylin-eosin (HE) staining. ResultsIn the CGHD-E acute toxicity experiment, the drug-treated groups did not show adverse reactions or poisoning symptoms, and the maximum tolerated dose of CGHD-E in mice was greater than 45.072 g/kg. In the long-term toxicity experiment, drug-treated rats generally exhibited a good condition, but continuous administration decreased on body weight and food intake, especially in male rats. Coagulation function alterations and the impact on the liver during long-term drug administration were also assessed, which should be emphasized in clinical applications. No significant toxic effects were observed according to routine blood tests or test of liver and kidney function, glucose and lipid metabolism, or ion metabolism. ConclusionsThe results of this study showed that CGHD was nontoxic or had low toxicity, providing not only a scientific basis for its clinical application, determining the appropriate clinical dose and monitoring clinical toxicity but also theoretical support for subsequent clinical drug trials.

The Cardiorenal Effects of Piper amalago Are Mediated by the Nitric Oxide/Cyclic Guanosine Monophosphate Pathway and the Voltage-Dependent Potassium Channels.

Piper amalago L. is used in Brazilian traditional medicine to treat inflammation, chest pain, and anxiety. This study aimed to investigate the safety and the renal and cardiovascular effects of the volatile oil (VO) and the aqueous (AE) and hydroalcoholic (HE) extracts from P. amalago. The gas chromatography-mass spectrometry analyses identified 47 compounds in the VO, with β-cyclogermacrene, spathulenol, β-phellandrene, and α-pinene standing out. Among the 47 compounds also found in AE and HE by liquid chromatography-mass spectrometry, glycosylated flavones, organic acids, amino acids, and amides were highlighted. Some examples of these compounds are methoxy-methylenedioxy cis-cinnamoyl pyrrolidine, methoxy-methylenedioxy trans-cinnamoyl pyrrolidine, and cyclobutene-2,4-bis-(1,3-benzodioxol-5-methoxy-6-yl)-1,3-dicarboxapyrrolidide. The acute toxicity experiments were conducted on female rats (n = 5). The cardiorenal assays (n = 8) and evaluations of vasodilatory effects on the mesenteric vascular bed (n = 5) were conducted on male rats. In either extract or VO, there were no mortality or changes in relative weights or histopathological analysis of the organs. Urinary volume and renal electrolyte excretion were elevated significantly during repeated dose 7-day treatment with different preparations from P. amalago. None of the preparations induced hypotension or changes in cardiac electrical activity. Only HE promoted significant vasodilatory effects in rats' isolated mesenteric vascular beds. These effects were completely abolished in the presence of L-NAME plus 4-aminopyridine. Therefore, P. amalago leaves are safe and present diuretic activity after acute and repeated dose administration over 7 days. Moreover, the HE induced significant vasodilator response in rats' mesenteric vascular beds by NO/cGMP pathway and voltage-dependent K+ channels activation.

Biodegradation of benzo(a)pyrene by a genetically engineered Bacillus licheniformis: Degradation, metabolic pathway and toxicity analysis

Benzo(a)pyrene (BaP), the polycyclic aromatic hydrocarbons (PAHs), is known for its high carcinogenic, low bioavailability, and resistance to removal. To enhance the removal efficiency of BaP, a screened BaP-dominant degrading bacterium identified as Bacillus licheniformis, was utilized to construct a genetically engineered Bacillus licheniformis, named GEM-NY2. Through successive subculturing, it was observed that the genetic stability of the first 20 generations of GEM-NY2 remained good, and the removal rate of BaP was 78.21–80.16 %. Compared to the recipient strain NY2, GEM-NY2 achieved a BaP removal rate of 78.61 %, showing a 10.36 % improvement. Then the optimal degradation parameters of the GEM-NY2 for BaP were investigated by the response surface methodology (RSM): salinity of 30 g/L, temperature of 37.5℃, pH of 6, and microbial inoculum of 3.5 %. Furthermore, UPLC-MS analysis confirmed that GEM-NY2 initiated the attack on BaP at sites C4 and C5, C7 and C8, and C9 and C10, leading to the formation of dihydroxy-BaP, then generating ring-cleavage products. Eight metabolites were obtained, and the ring-opening process of BaP by GEM-NY2 was revealed. The results of EcoSAR and acute toxicity experiments of Photobacterium phosphoreum showed as the benzene rings of BaP were cleaved, the toxicity of the metabolites gradually decreased. The luminescence inhibition rate decreased significantly from 82.89 % to 52.36 % within 14 days. In summary, the genetically engineered Bacillus licheniformis (GEM-NY2) constructed in this work exhibited promising bio-degradation potential towards BaP, highlighting its favorable prospects for the effective removal of PAHs.

Design and synthesis of pterostilbene derivatives bearing triazole moiety that might treat DSS-induced colitis in mice through modulation of NF-κB/MAPK signaling pathways

In this study, a series of novel anti-inflammatory compounds with high activity and low toxicity were designed and synthesized based on the natural product pterostilbene skeleton. According to the strategy of pharmacophore combination, we introduced thiazole moiety into pterostilbene skeleton to design and synthesize a novel series of pterostilbene derivatives (a total of 41 compounds), and lipopolysaccharide (LPS)-treated RAW 264.7 cells were screened for anti-inflammatory activity and cytotoxicity. Among them, compound 8 was found to be the most active (against NO: IC50 = 0.6 μM) compared with pterostilbene and indomethacin. Anti-inflammatory mechanism studies revealed that compound 8 inhibited pro-inflammatory cytokines by blocking the NF-κB/MAPK signaling pathway in LPS-treated RAW 264.7 cells. In vivo experiments showed that compound 8 had a good relieving effect on DSS-induced acute colitis in mice, and also demonstrated a good safety in acute toxicity experiments. In conclusion, compound 8 may be a promising anti-inflammatory lead compound in the treatment of acute colitis.

Electrical Stimulation of Vocal Fold Adduction Triggered by Laryngeal Electromyography Using a Custom Implant.

Medialization procedures for unilateral vocal fold (VF) paralysis generally improve voice but do not fully replace dynamic VF adduction. Paralyzed VFs typically experience synkinetic reinnervation, which makes it feasible to elicit movement through electrical stimulation. We tested a novel laryngeal pacing implant capable of providing closed-loop (automatic) stimulation of a VF triggered by electromyography (EMG) potentials from the contralateral VF. A custom, battery-powered, microprocessor-based stimulator was tested in eight dogs with bipolar electrodes implanted for recording EMG from the left VF and stimulating adduction of the right VF. A cuff electrode on the left recurrent laryngeal nerve (RLN) stimulated unilateral VF adduction, modeling voluntary control in anesthetized animals. Closed-loop stimulation was tested in both acute and chronic experiments. Synkinetic reinnervation was created in two animals by right RLN transection and suture repair to model unilateral VF paralysis. In all animals, left VF activation through RLN stimulation generated a robust EMG response that rapidly triggered stimulation of contralateral thyroarytenoid and lateral cricoarytenoid muscles, causing nearly simultaneous bilateral adduction. Optimal triggering of VF stimulation from elicited EMG was achieved using independent onset and offset thresholds. Real-time artifact blanking allowed closed-loop stimulation without self-perpetuating feedback, despite the proximity of recording and stimulation electrodes. Using a custom implant system, we demonstrated real-time closed-loop stimulation of one VF triggered by the activation of the contralateral VF. This approach could potentially restore dynamic glottic closure for reflexive behaviors or phonation in cases of unilateral VF paralysis with synkinetic reinnervation. https://doi.org/10.23641/asha.24492133.

Sodium butyrate improves the growth performance, intestinal immunity, and ammonia tolerance of yellow catfish Pelteobagrus fulvidraco by increasing the relative abundance of Candidatus_Arthromitus

The study aimed to analyze the effect of dietary sodium butyrate (SB) on intestinal digestive enzyme activity, liver physiological status, immune parameter, intestinal microbiota structure and composition, and ammonia tolerance in yellow catfish. Over the course of an 8-week experiment, four separate groups including 480 healthy male yellow catfish (1.52 ± 0.66 g) were fed diets containing varying concentrations of SB: 0, 0.05, 0.1, and 0.2%. Following this, an acute ammonia stress experiment that lasted for 96-h was conducted. The study showed that dietary supplementation of SB can improve growth performance and feed utilization (weight gain rate increased from 163.66 to 226.1%; specific growth rate increased from 1.73 to 2.11%/d; feed conversion rate decreased from 2.02 to 1.34), and increase intestinal digestive enzyme activity (lipase increased from 22.66 to 41.46 U/mg prot; amylase increased from 0.38 to 0.46 U/mg prot; pepsin increased from 10.13 to 12.83 U/mg prot). Plasma total protein (from 510.79 to 523.02 μg/mL), 50% hemolytic complement (from 35.46 to 40.46 U/mL), and immunoglobulin M (from 0.45 to 0.48 g/L) increased as dietary SB levels. On the contrary, plasma glucose (from 6.2 to 3.39 mmol/mL), high-density lipoprotein cholesterol (from 0.38 to 0.25 mmol/mL), aspartate aminotransferase (from 55.66 to 37.36 U/L), alanine aminotransferase (from 358.82 to 283.25 U/L), superoxide dismutase (from 163.84 to 147.13 U/mL), catalase (from 152.89 to 135.07 U/mL), tumor necrosis factor-α (from 67.58 to 62.13 pg/mL), interleukin-1 (from 40.07 to 30.96 pg/mL), and interleukin-8 (from 57.16 to 52.16 pg/mL) decreased with dietary SB levels. In addition, dietary supplementation of 0.2% SB significantly increased the relative abundance of Candidatus_Arthromitus in intestine by approximately 22-fold. To sum up, the utilization of SB has potential to enhance fish growth performance, elevate its physiological status, and improve its ability to tolerate acute ammonia toxicity. This is achievable by increasing the prevalence of Candidatus_Arthromitus in the intestine.