Showing papers in "Physiological Reports in 2023"

Electrophysiological activity from over the cerebellum and cerebrum during eye blink conditioning in human subjects

Abstract: We report the results of an experiment in which electrophysiological activity was recorded from the human cerebellum and cerebrum in a sample of 14 healthy subjects before, during and after a classical eye blink conditioning procedure with an auditory tone as conditional stimulus and a maxillary nerve unconditional stimulus. The primary aim was to show changes in the cerebellum and cerebrum correlated with behavioral ocular responses. Electrodes recorded EMG and EOG at peri‐ocular sites, EEG from over the frontal eye‐fields and the electrocerebellogram (ECeG) from over the posterior fossa. Of the 14 subjects half strongly conditioned while the other half were resistant. We confirmed that conditionability was linked under our conditions to the personality dimension of extraversion‐introversion. Inhibition of cerebellar activity was shown prior to the conditioned response, as predicted by Albus (1971). However, pausing in high frequency ECeG and the appearance of a contingent negative variation (CNV) in both central leads occurred in all subjects. These led us to conclude that while conditioned cerebellar pausing may be necessary, it is not sufficient alone to produce overt behavioral conditioning, implying the existence of another central mechanism. The outcomes of this experiment indicate the potential value of the noninvasive electrophysiology of the cerebellum.

Aging affects the number and morphological heterogeneity of rat phrenic motor neurons and phrenic motor axons

Abstract: Diaphragm muscle (DIAm) motor units comprise a phrenic motor neuron (PhMN), the phrenic nerve and the muscle fibers innervated, with the size of PhMNs and axons characteristic of motor unit type. Smaller PhMNs and their axons comprise slow (type S) and fatigue‐resistant (type FR) DIAm motor units, while larger PhMNs and their axons comprise more fatigable (type FF) motor units. With aging, we have shown a loss of larger PhMNs, consistent with selective atrophy of type IIx/IIb DIAm fibers and reduced maximum DIAm force. In the present study, we hypothesized that with aging there is a loss of larger myelinated phrenic α motor axons. Female and male young (6 months) and old (24 months) Fischer 344 rats were studied. PhMNs were retrogradely labeled by intrapleural injection of 488‐conjugated CTB. The phrenic nerves were excised ~1 cm from the DIAm insertion and mounted in resin, and phrenic α motor axons were delineated based on size (i.e., >4 μm diameters). In older rats, the number of larger PhMNs and larger phrenic α motor axons were reduced. There were no differences in non‐α axons. In addition, there was evidence of demyelination of larger phrenic α motor axons in older rats. Together, these findings are consistent with the selective age‐related vulnerability of larger PhMNs and denervation of type FF motor units, which may underlie DIAm sarcopenia.

Validation of commercially available antibodies directed against subunits of the epithelial Na+ channel

Abstract: The epithelial Na+ channel (ENaC) is traditionally composed of three subunits, although non‐canonical expression has been found in various tissues including the vasculature, brain, lung, and dendritic cells of the immune system. Studies of ENaC structure and function have largely relied on heterologous expression systems, often with epitope‐tagged channel subunits. Relevant in vivo physiological studies have used ENaC inhibitors, mice with global or tissue specific knockout of subunits, and anti‐ENaC subunit antibodies generated by investigators or by commercial sources. Availability of well‐characterized, specific antibodies is imperative as we move forward in understanding the role of ENaC in non‐epithelial tissues where expression, subunit organization, and electrophysiological characteristics may differ from epithelial tissues. We report that a commonly used commercial anti‐α subunit antibody recognizes an intense non‐specific band on mouse whole kidney and lung immunoblots, which migrates adjacent to a less intense, aldosterone‐induced full length α‐subunit. This antibody localizes to the basolateral membrane of aquaporin 2 negative cells in kidney medulla. We validated antibodies against the β‐ and γ‐subunits from the same commercial source. Our work illustrates the importance of validation studies when using popular, commercially available anti‐ENaC antibodies.

Caveolin‐1 inhibition mediates the opposing effects of alcohol on γ‐secretase activity in arterial endothelial and smooth muscle cells

Abstract: Notch is important to vessel homeostasis. We investigated the mechanistic role of caveolin‐1 (Cav‐1) in mediating the effects of alcohol (Ethanol/EtOH) on the γ‐secretase proteolytic activity necessary for Notch signaling in vascular cells. Human coronary artery endothelial cells (HCAEC) were treated with EtOH (0–50 mM), Notch ligand delta‐like ligand 4 (Dll4), and the γ‐secretase inhibitor DAPT. EtOH stimulated Notch signaling in HCAEC as evidenced by increased Notch receptor (N1, N4) and target gene (hrt2, hrt3) mRNA levels with the most robust response achieved at 25 mM EtOH. Ethanol (25 mM) stimulated γ‐secretase proteolytic activity, to the same extent as Dll4, in HCAEC membranes. Ethanol inhibited Cav‐1 mRNA and protein levels in HCAEC. Caveolin‐1 negatively regulated γ‐secretase activity in HCAEC as Cav‐1 knockdown stimulated it, while Cav‐1 overexpression inhibited it. Moreover, Cav‐1 overexpression blocked the stimulatory effect of EtOH on γ‐secretase activity in HCAEC. Although EtOH also inhibited Cav‐1 expression in human coronary artery smooth muscle cells (HCASMC), EtOH inhibited γ‐secretase activity in HCASMC in contrast to its effect in HCAEC. The inhibitory effect of EtOH on γ‐secretase in HCASMC was mimicked by Cav‐1 knockdown and prevented by Cav‐1 overexpression, suggesting that in these cells Cav‐1 positively regulates γ‐secretase activity. In conclusion, EtOH differentially regulates γ‐secretase activity in arterial EC and SMC, being stimulatory and inhibitory, respectively. These effects are both mediated by caveolin‐1 inhibition which itself has opposite effects on γ‐secretase in the two cell types. This mechanism may underlie, in part, the effects of moderate drinking on atherosclerosis.

Vagus nerve stimulation parameters evoke differential neuronal responses in the locus coeruleus

Abstract: Vagus nerve stimulation (VNS) is used to treat drug‐resistant epilepsy and depression, with additional applications under investigation. The noradrenergic center locus coeruleus (LC) is vital for VNS effects; however, the impact of varying stimulation parameters on LC activation is poorly understood. This study characterized LC activation across VNS parameters. Extracellular activity was recorded in rats' left LC while 11 VNS paradigms, utilizing variable frequencies and bursting characteristics, were pseudorandomly delivered to the left cervical vagus for five cycles. Neurons' change from baseline firing rate and timing response profiles were assessed. The proportion of neurons categorized as responders over 5 VNS cycles doubled in comparison to the first VNS cycle (p < 0.001) for all VNS paradigms, demonstrating an amplification effect. The percentage of positively consistent/positive responders increased for standard VNS paradigms with frequencies ≥10 Hz and for bursting paradigms with shorter interburst intervals and more pulses per burst. The synchrony between pairs of LC neurons increased during bursting VNS but not standard paradigms. Also, the probability of evoking a direct response during bursting VNS was higher with longer interburst intervals and a higher number of pulses per burst. Standard paradigms between 10–30 Hz best positively activates LC with consistency to VNS while the best bursting paradigm to increase activity was 300 Hz, seven pulses per burst separated by 1 s. Bursting VNS was effective in increasing synchrony between pairs of neurons, suggesting a common network recruitment originating from vagal afferents. These results indicate differential activation of LC neurons depending on the VNS parameters delivered.

Characterization of acute lung injury in the bleomycin rat model

Abstract: The aim of this study was to describe and characterize the pathophysiological changes occurring during the early inflammatory phase (first 3 days) in the rat bleomycin model of lung injury preceding the development of fibrosis. Further, we wanted to understand the kinetics and factors contributing to bleomycin‐induced acute lung injury (ALI) and provide a robust, reliable and reproducible framework of features of ALI readouts to assess effects of therapeutics on bleomycin‐induced ALI in rats. We induced ALI in rats with intratracheal (i.t.) installation of bleomycin. The animals were sacrificed on predetermined time points, that is, Day 0, 1, 2, and 3 post the bleomycin challenge. We analyzed bronchoalveolar lavage fluid (BALF) and lung tissue to establish and assess relevant experimental features of ALI. We demonstrated that bleomycin induced key features of experimental ALI including a profound increase in neutrophils in BALF (50–60%), pulmonary edema, and lung pathology on Day 3 after challenge. Furthermore, we showed that TGF‐β1, IL‐1β, TNF‐α, IL‐6, CINC‐1, TIMP‐1, and WISP‐1 were induced by studying their kinetic profile during the first 3 days after bleomycin injury consistent with their known role ALI. We also confirmed that detectable fibrogenesis occurs at the earliest on Day 3 after injury based on collagen content, along with changes in the TGF‐β/Smad signaling pathway and increased expression of Galectin‐3, Vimentin, and Fibronectin in lung homogenate. Our report presents robust features and contributing mediators/factors to the pathology of bleomycin‐induced ALI in rats on Day 3. The kinetic data provide insights on the progression of ALI and a detailed understanding of early events before actual fibrosis development. This set of experimental endpoints is very appropriate and invaluable for efficacy testing of potential novel therapeutic treatments (single or combined) in ALI and understanding their mechanism of action.

Functional roles of CD26/DPP4 in bleomycin‐induced pulmonary fibrosis

Abstract: The pathogenesis of pulmonary fibrosis involves complex interplay between cell types and signaling pathways. Recurrent alveolar epithelial injury can occur during pulmonary inflammation, causing dysregulation of epithelial repair. Dysregulated repair interacts with mesenchymal, inflammatory, and endothelial cells to trigger fibroblast‐to‐myofibroblast activation. CD26/dipeptidyl peptidase‐4 (DPP4) is a type II membrane protein mediating pleiotropic effect. However, the mechanistic role of CD26/DPP4 in pulmonary fibrosis remains unclear. In this study, we aimed to characterize Dpp4 deficiency in a mouse bleomycin (BLM)‐induced pulmonary fibrosis model and in cell culture systems of human lung fibroblasts (HLFs). Dpp4 knockout (Dpp4 KO) mouse lungs exhibited lower Ashcroft scale indices, collagen content, and numbers of fibroblasts and myofibroblasts compared with those in C57BL/6 wild‐type (WT) mice. Upregulation of Tgfb1 and Tgfb2 mRNA levels in the lungs after BLM treatment was lower in Dpp4 KO mice compared with those in WT mice. Although TGF‐β‐driven endothelial‐to‐mesenchymal transition (EndMT) has been implicated as one of the mechanisms of pulmonary fibrosis, a number of partial EndMT cells in lungs did not differ between Dpp4 KO mice and WT mice. The proliferation capacity and mRNA levels of COL1A1, a collagen deposition‐related gene, in cultured HLFs were suppressed in DPP4 small interfering RNA‐treated cells. This study indicates that the genetic deficiency of DPP4 has protective effects against BLM‐induced pulmonary fibrosis, partly through the reduction in TGF‐β expression and inhibition of fibroblast activation in the lung. Our study suggests that CD26/DPP4 inhibition is a potential therapeutic strategy for pulmonary fibrosis.

Characteristics of acid‐sensing ion channel currents in male rat muscle dorsal root ganglion neurons following ischemia/reperfusion

Abstract: Peripheral artery diseases (PAD) increases muscle afferent nerve‐activated reflex sympathetic nervous and blood pressure responses during exercise (termed as exercise pressor reflex). However, the precise signaling pathways leading to the exaggerated autonomic responses in PAD are undetermined. Considering that limb ischemia/reperfusion (I/R) is a feature of PAD, we determined the characteristics of acid‐sensing ion channel (ASIC) currents in muscle dorsal root ganglion (DRG) neurons under the conditions of hindlimb I/R and ischemia of PAD. In particular, we examined ASIC currents in two distinct subpopulations, isolectin B4‐positive, and B4‐negative (IB4+ and IB4−) muscle DRG neurons, linking to glial cell line‐derived neurotrophic factor and nerve growth factor. In results, ASIC1a‐ and ASIC3‐like currents were observed in IB4− muscle DRG neurons with a greater percentage of ASIC3‐like currents. Hindimb I/R and ischemia did not alter the distribution of ASIC1a and ASIC3 currents with activation of pH 6.7 in IB4+ and IB4− muscle DRG neurons; however, I/R altered the distribution of ASIC3 currents in IB4+ muscle DRG neurons with pH 5.5, but not in IB4− neurons. In addition, I/R and ischemia amplified the density of ASIC3‐like currents in IB4− muscle DRG neurons. Our results suggest that a selective subpopulation of muscle afferent nerves should be taken into consideration when ASIC signaling pathways are studied to determine the exercise pressor reflex in PAD.

Mechanical energy on anaerobic capacity during a supramaximal treadmill running in men: Is there influence between runners and active individuals?

Abstract: This study verified whether mechanical variables influence the anaerobic capacity outcome on treadmill running and whether these likely influences were dependent of running experience. Seventeen physical active and 18 amateur runners, males, performed a graded exercise test and constant load exhaustive running efforts at 115% of intensity associated to maximal oxygen consumption. During the constant load were determined the metabolic responses (i.e., gas exchange and blood lactate) to estimate the energetic contribution and anaerobic capacity as well as kinematic responses. The runners showed higher anaerobic capacity (16.6%; p = 0.005), but lesser time to exercise failure (−18.8%; p = 0.03) than active subjects. In addition, the stride length (21.4%; p = 0.00001), contact phase duration (−11.3%; p = 0.005), and vertical work (−29.9%; p = 0.015). For actives, the anaerobic capacity did not correlate significantly with any physiologic, kinematic, and mechanical variables and no regression model was fitted using the stepwise multiple regression, while to runners the anaerobic capacity was significantly correlated with phosphagen energetic contribution (r = 0.47; p = 0.047), external power (r = −0.51; p = 0.031), total work (r = −0.54; p = 0.020), external work (r = −0.62; p = 0.006), vertical work (r = −0.63; p = 0.008), and horizontal work (r = −0.61; p = 0.008), and the vertical work and phosphagen energetic contribution presented a coefficient of determination of 62% (p = 0.001). Based on findings, it is possible to assume that for active subjects, the mechanical variables have no influence over the anaerobic capacity, however, for experienced runners, the vertical work and phosphagen energetic contribution have relevant effect over anaerobic capacity output.

Exercise training reduces circulating cytokines in male patients with coronary artery disease and type 2 diabetes: A pilot study

Abstract: Low‐grade inflammation is central to coronary artery disease (CAD) and type 2 diabetes (T2D) and is reduced by exercise training. The objective of this study was to compare the anti‐inflammatory potential of moderate‐to‐vigorous intensity continuous training (MICT) and high‐intensity interval training (HIIT) in patients with CAD with or without T2D. The design and setting of this study is based on a secondary analysis of registered randomized clinical trial NCT02765568. Male patients with CAD were randomly assigned to either MICT or HIIT, with subgroups divided according to T2D status (non‐T2D‐HIIT n = 14 and non‐T2D‐MICT n = 13; T2D‐HIIT n = 6 and T2D‐MICT n = 5). The intervention was a 12‐week cardiovascular rehabilitation program consisting of either MICT or HIIT (twice weekly sessions) and circulating cytokines measured pre‐ and post‐training as inflammatory markers. The co‐occurrence of CAD and T2D was associated with increased plasma IL‐8 (p = 0.0331). There was an interaction between T2D and the effect of the training interventions on plasma FGF21 (p = 0.0368) and IL‐6 (p = 0.0385), which were further reduced in the T2D groups. An interaction between T2D, training modalities, and the effect of time (p = 0.0415) was detected for SPARC, with HIIT increasing circulating concentrations in the control group, while lowering them in the T2D group, and the inverse occurring with MICT. The interventions also reduced plasma FGF21 (p = 0.0030), IL‐6 (p = 0.0101), IL‐8 (p = 0.0087), IL‐10 (p < 0.0001), and IL‐18 (p = 0.0009) irrespective of training modality or T2D status. HIIT and MICT resulted in similar reductions in circulating cytokines known to be increased in the context of low‐grade inflammation in CAD patients, an effect more pronounced in patients with T2D for FGF21 and IL‐6.

Boric acid transport activity of marine teleost aquaporins expressed in Xenopus oocytes

Abstract: Marine teleosts ingest large amounts of seawater containing various ions, including 0.4 mM boric acid, which can accumulate at toxic levels in the body. However, the molecular mechanisms by which marine teleosts absorb and excrete boric acid are not well understood. Aquaporins (Aqps) are homologous to the nodulin‐like intrinsic protein (NIP) family of plant boric acid channels. To investigate the potential roles of Aqps on boric acid transport across the plasma membrane in marine teleosts, we analyzed the function of Aqps of Japanese pufferfish (Takifugu rubripes) expressed in Xenopus laevis oocytes. Takifugu genome database contains 16 genes encoding the aquaporin family members (aqp0a, aqp0b, aqp1aa, aqp1ab, aqp3a, aqp4a, aqp7, aqp8bb, aqp9a, aqp9b, aqp10aa, aqp10bb, aqp11a, aqp11b, aqp12, and aqp14). When T. rubripes Aqps (TrAqps) were expressed in X. laevis oocytes, a swelling assay showed that boric acid permeability was significantly increased in oocytes expressing TrAqp3a, 7, 8bb, 9a, and 9b. The influx of boric acid into these oocytes was also confirmed by elemental quantification. Electrophysiological analysis using a pH microelectrode showed that these TrAqps increase B(OH)3 permeability. These results indicate that TrAqp3a, 7, 8bb, 9a, and 9b act as boric acid transport systems, likely as channels, in marine teleosts.

A role for β‐catenin in diet‐induced skeletal muscle insulin resistance

Abstract: A central characteristic of insulin resistance is the impaired ability for insulin to stimulate glucose uptake into skeletal muscle. While insulin resistance can occur distal to the canonical insulin receptor‐PI3k‐Akt signaling pathway, the signaling intermediates involved in the dysfunction are yet to be fully elucidated. β‐catenin is an emerging distal regulator of skeletal muscle and adipocyte insulin‐stimulated GLUT4 trafficking. Here, we investigate its role in skeletal muscle insulin resistance. Short‐term (5‐week) high‐fat diet (HFD) decreased skeletal muscle β‐catenin protein expression 27% (p = 0.03), and perturbed insulin‐stimulated β‐cateninS552 phosphorylation 21% (p = 0.009) without affecting insulin‐stimulated Akt phosphorylation relative to chow‐fed controls. Under chow conditions, mice with muscle‐specific β‐catenin deletion had impaired insulin responsiveness, whereas under HFD, both mice exhibited similar levels of insulin resistance (interaction effect of genotype × diet p < 0.05). Treatment of L6‐GLUT4‐myc myocytes with palmitate lower β‐catenin protein expression by 75% (p = 0.02), and attenuated insulin‐stimulated β‐catenin phosphorylationS552 and actin remodeling (interaction effect of insulin × palmitate p < 0.05). Finally, β‐cateninS552 phosphorylation was 45% lower in muscle biopsies from men with type 2 diabetes while total β‐catenin expression was unchanged. These findings suggest that β‐catenin dysfunction is associated with the development of insulin resistance.

Key role for Kv11.1 (ether‐a‐go‐go related gene) channels in rat bladder contractility

Abstract: In addition, to their established role in cardiac myocytes and neurons, ion channels encoded by ether‐a‐go‐go‐related genes (ERG1‐3 or kcnh2,3 and 6) (kcnh2) are functionally relevant in phasic smooth muscle. The aim of the study was to determine the expression and functional impact of ERG expression products in rat urinary bladder smooth muscle using quantitative polymerase chain reaction, immunocytochemistry, whole‐cell patch‐clamp and isometric tension recording. kcnh2 was expressed in rat bladder, whereas kcnh6 and kcnh3 expression were negligible. Immunofluorescence for the kcnh2 expression product Kv11.1 was detected in the membrane of isolated smooth muscle cells. Potassium currents with voltage‐dependent characteristics consistent with Kv11.1 channels and sensitive to the specific blocker E4031 (1 μM) were recorded from isolated detrusor smooth muscles. Disabling Kv11.1 activity with specific blockers (E4031 and dofetilide, 0.2–20 μM) augmented spontaneous contractions to a greater extent than BKCa channel blockers, enhanced carbachol‐driven activity, increased nerve stimulation‐mediated contractions, and impaired β‐adrenoceptor‐mediated inhibitory responses. These data establish for the first time that Kv11.1 channels are key determinants of contractility in rat detrusor smooth muscle.

The discovery of a new type of innervation in lymphoid organs

Abstract: It is well known that the main forms of innervation are synapses and free nerve endings, while other forms of innervation have not been reported. Here, we explore a new way of innervating lymphoid organs. Male Sprague‐Dawley rats were used for studying the innervation of sympathetic nerve fibers in lymph nodes by means of anterograde tracking, immunoelectron microscopy, three‐dimension reconstruction analysis, and immunofluorescence labeling. The results showed that the Fluoro‐Ruby labeled nerve endings targeted only a group of cells in the lymph nodes and entered the cells through the plasma membrane. The electron microscopy showed that the biotinylated glucan amine reaction elements were distributed in the cytoplasm, and most of the biotinylated glucan amine active elements were concentrated on the microtubule and microfilament walls. Birbeck particles with rod‐shaped and/or tennis racket like structures can be seen in the labeled cells at high magnification, and Birbeck particles contain biotinylated glucan amine‐reactive elements. The immunofluoresence results showed that the Fluoro‐Ruby‐labeled nerve innervating cells expressed CD207 and CD1a protein. This result confirmed that the labeled cells were Langerhans cells. Our findings suggested that Langerhans cells might serve as a “bridge cell” for neuroimmune cross‐talking in lymph organs, which play an important role in transmitting signals of the nervous system to immune system. This study also opened up a new way for further study of immune regulation mechanism.

Validity and reproducibility of a whole‐room indirect calorimeter for the estimation of VO2 , VCO2 , and resting metabolic rate

Abstract: Whole‐room indirect calorimeters (WRICs) provide accurate instruments for the measurement of respiratory exchange, energy expenditure, and macronutrient oxidation. Here, we aimed to determine the validity and reproducibility of a 7500 L WRIC for the measurement of ventilation rates and resting metabolic rate (RMR). Technical validation was performed with propane combustion tests (n = 10) whereas biological reproducibility was tested in healthy subjects (13 women, 6 men, mean ± SD age 39.6 ± 15.3) in two 60 min measurements separated by 24 h. Subjects followed a run‐in protocol prior to measurements. The coefficient of variation (CV) and intraclass correlation coefficient (ICC) were calculated for ventilation rates of O2 (VO2), CO2 (VCO2), the respiratory quotient (RQ; VCO2/VO2), and RMR. Technical validation showed good validity with CVs ranging from 0.67% for VO2 to 1.00% for energy expenditure. For biological reproducibility, CVs were 2.89% for VO2; 2.67% for VCO2; 1.95% for RQ; and 2.68% for RMR. With the exception of RQ (74%), ICCs were excellent for VO2 (94%), VCO2 (96%) and RMR (95%). Excluding participants that deviated from the run‐in protocol did not alter results. In conclusion, the 7500 L WRIC is technically valid and reproducible for ventilation rates and RMR.

Method for estimating pulsatile wall shear stress from one‐dimensional velocity waveforms

Abstract: Wall shear stress (WSS)—a key regulator of endothelial function—is commonly estimated in vivo using simplified mathematical models based on Poiseuille's flow, assuming a quasi‐steady parabolic velocity distribution, despite evidence that more rapidly time‐varying, pulsatile blood flow during each cardiac cycle modulates flow‐mediated dilation (FMD) in large arteries of healthy subjects. More exact and accurate models based on the well‐established Womersley solution for rapidly changing blood flow have not been adopted clinically, potentially because the Womersley solution relies on the local pressure gradient, which is difficult to measure non‐invasively. We have developed an open‐source method for automatic reconstruction of unsteady, Womersley‐derived velocity profiles, and WSS in conduit arteries. The proposed method (available online at https://doi.org/10.5281/zenodo.7576408) requires only the time‐averaged diameter of the vessel and time‐varying velocity data available from non‐invasive imaging such as Doppler ultrasound. Validation of the method with subject‐specific computational fluid dynamics and application to synthetic velocity waveforms in the common carotid, brachial, and femoral arteries reveals that the Poiseuille solution underestimates peak WSS 38.5%–55.1% during the acceleration and deceleration phases of systole and underestimates or neglects retrograde WSS. Following evidence that oscillatory shear significantly augments vasodilator production, it is plausible that mischaracterization of the shear stimulus by assuming parabolic flow leads to systematic underestimates of important biological effects of time‐varying blood velocity in conduit arteries.

Profound hyponatremia and dehydration: A case of cisplatin induced renal salt wasting syndrome

Abstract: Cisplatin is a well‐known chemotherapeutic agent that can be associated with hyponatremia. It is known to be associated with a multitude of renal disorders including acute kidney injury with reduced glomerular filtration, Fanconi syndrome, and renal tubular acidosis, nephrogenic diabetes insipidus and renal salt wasting syndrome. We report a case of an elderly male presenting with significant recurrent hyponatremia, and prerenal azotemia. With recent exposure to cisplatin along with significant hypovolemia and urinary loss of sodium, he was diagnosed to have cisplatin induced renal salt wasting syndrome.

Rapid adjustments to autonomic control of cardiac rhythm at the onset of isometric exercise in healthy young adults

Abstract: Sympathetic nervous system (SNS) and parasympathetic nervous system (PNS) influences on cardiac rhythm at the onset of exercise, a time of rapid autonomic adjustments, are clinically important areas of investigation. Continuous wavelet transform (CWT) involves time‐frequency‐based heart rate variability (HRV) analysis allowing investigation of autonomic influences on cardiac rhythm during short durations of exercise. Therefore, the purpose of this study was to characterize SNS and PNS influences on cardiac rhythm at the onset of isometric exercise in healthy young adults. CWT analysis was retrospectively applied to R‐R interval data (electrocardiogram) previously collected from 14 healthy young adults (26 ± 2 years) who performed 30‐s, one‐legged, isometric, calf exercise at 70% maximal voluntary contraction (MVC; 70% MVC trial) or rested (0% MVC trial). Absolute and normalized low‐frequency (aLF, nLF; 0.04–0.15 Hz) and high‐frequency (aHF, nHF; 0.15–0.4 Hz) bands and LF/HF were used to analyze one 30‐s baseline period and six 5‐s time windows during the 30‐s exercise (70% MVC) or rest (0% MVC). Statistical analysis involved two‐way analysis of variance with post‐hoc analysis. aHF, aLF, LF/HF, nHF, and nLF displayed a trial‐time interaction (all p ≤ 0.027). In the 70% compared to the 0% MVC trial, aHF and nHF were lower after 5–30 s (all p ≤ 0.040), aLF was lower after 20–30 s (all p ≤ 0.011) and LF/HF and nLF were higher after 5–20 s (all p ≤ 0.045). These results indicate the reduction of the PNS influence on cardiac rhythm begins sooner than the augmentation of the SNS influence at the onset of isometric exercise in healthy young adults.

Effects of beetroot juice supplementation on vascular functional and structural changes in aged mice

Abstract: This study investigated whether beetroot juice (BRJ) ingestion ameliorates aging‐induced functional and structural changes in vasculature. Aged mice (98–100 weeks old) were supplemented with BRJ (nitrate: 3.5 mmol/L) or drinking water for 4 weeks and compared with young mice (12–15 weeks old). The vasorelaxant response of isolated aortas to acetylcholine was markedly weaker in aged mice than in young mice, but the attenuated relaxation was significantly improved in BRJ‐supplemented aged mice. The acetylcholine‐induced relaxation was completely abolished by Nω‐nitro‐l‐arginine methyl ester in all groups. Additionally, the response to sodium nitroprusside was comparable among the three groups. The aortic medial thickness was significantly greater in aged mice than in young mice, and BRJ supplementation did not suppress this thickening. Plasma nitrate levels were significantly higher in BRJ‐supplemented aged mice than in non‐supplemented aged mice. Conversely, non‐supplemented aged mice had high plasma levels of thiobarbituric acid‐reactive substances, but the levels were suppressed in BRJ‐supplemented aged mice. These findings suggest that BRJ ingestion improves vascular endothelial dysfunction associated with aging, at least in part, by enhancing nitric oxide bioavailability and reducing oxidative stress. Therefore, beetroot ingestion may be a highly useful self‐medication option to prevent vascular aging.

Allometric scaling of metabolic rate and cardiorespiratory variables in aquatic and terrestrial mammals

Abstract: While basal metabolic rate (BMR) scales proportionally with body mass (Mb), it remains unclear whether the relationship differs between mammals from aquatic and terrestrial habitats. We hypothesized that differences in BMR allometry would be reflected in similar differences in scaling of O2 delivery pathways through the cardiorespiratory system. We performed a comparative analysis of BMR across 63 mammalian species (20 aquatic, 43 terrestrial) with a Mb range from 10 kg to 5318 kg. Our results revealed elevated BMRs in small (>10 kg and <100 kg) aquatic mammals compared to small terrestrial mammals. The results demonstrated that minute ventilation, that is, tidal volume (VT)·breathing frequency (fR), as well as cardiac output, that is, stroke volume·heart rate, do not differ between the two habitats. We found that the “aquatic breathing strategy”, characterized by higher VT and lower fR resulting in a more effective gas exchange, and by elevated blood hemoglobin concentrations resulting in a higher volume of O2 for the same volume of blood, supported elevated metabolic requirements in aquatic mammals. The results from this study provide a possible explanation of how differences in gas exchange may serve energy demands in aquatic versus terrestrial mammals.

Short‐term semaglutide treatment improves FGF21 responsiveness in primary hepatocytes isolated from high fat diet challenged mice

Abstract: Metabolic functions of GLP‐1 and its analogues have been extensively investigated. In addition to acting as an incretin and reducing body weight, we and others have suggested the existence of GLP‐1/fibroblast growth factor 21 (FGF21) axis in which liver mediates certain functions of GLP‐1 receptor agonists. In a more recent study, we found with surprise that four‐week treatment with liraglutide but not semaglutide stimulated hepatic FGF21 expression in HFD‐challenged mice. We wondered whether semaglutide can also improve FGF21 sensitivity or responsiveness and hence triggers the feedback loop in attenuating its stimulation on hepatic FGF21 expression after a long‐term treatment. Here, we assessed effect of daily semaglutide treatment in HFD‐fed mice for 7 days. HFD challenge attenuated effect of FGF21 treatment on its downstream events in mouse primary hepatocytes, which can be restored by 7‐day semaglutide treatment. In mouse liver, 7‐day semaglutide treatment stimulated FGF21 as well as genes that encode its receptor (FGFR1) and the obligatory co‐receptor (KLB), and a battery of genes that are involved in lipid homeostasis. In epididymal fat tissue, expressions of a battery genes including Klb affected by HFD challenge were reversed by 7‐day semaglutide treatment. We suggest that semaglutide treatment improves FGF21 sensitivity which is attenuated by HFD challenge.

Bioelectrical impedance phase angle is associated with physical performance before but not after simulated multi‐stressor military operations

Abstract: Physical performance decrements observed during multi‐stressor military operations may be attributed, in part, to cellular membrane dysfunction, which is quantifiable using phase angle (PhA) derived from bioelectrical impedance analysis (BIA). Positive relationships between PhA and performance have been previously reported in cross‐sectional studies and following longitudinal exercise training programs, but whether changes in PhA are indicative of acute decrements in performance during military operations is unknown. Data from the Optimizing Performance for Soldiers II study, a clinical trial examining the effects of exogenous testosterone administration on body composition and performance during military stress, was used to evaluate changes in PhA and their associations with physical performance. Recreationally active, healthy males (n = 34; 26.6 ± 4.3 years; 77.9 ± 12.4 kg) were randomized to receive testosterone undecanoate or placebo before a 20‐day simulated military operation, which was followed by a 23‐day recovery period. PhA of the whole‐body (Whole) and legs (Legs) and physical performance were measured before (PRE) and after (POST) the simulated military operation as well as in recovery (REC). Independent of treatment, PhAWhole and PhALegs decreased from PRE to POST (p < 0.001), and PhALegs, but not PhAWhole, remained lower at REC than PRE. PhAWhole at PRE and REC were associated with vertical jump height and Wingate peak power (p < 0.001–0.050), and PhAWhole at PRE was also associated with 3‐RM deadlift mass (p = 0.006). However, PhA at POST and changes in PhA from PRE to POST were not correlated with any performance measure (p > 0.05). Additionally, PhA was not associated with aerobic performance at any timepoint. In conclusion, reduced PhA from PRE to POST provides indirect evidence of cellular membrane disruption. Associations between PhA and strength and power were only evident at PRE and REC, suggesting PhA may be a useful indicator of strength and power, but not aerobic capacity, in non‐stressed conditions, and not a reliable indicator of physical performance during severe physiological stress.

Caffeine ingestion stimulates plasma carnitine clearance in humans

Abstract: Increasing skeletal muscle carnitine content can manipulate fuel metabolism and improve exercise performance. Intravenous insulin infusion during hypercarnitinemia increases plasma carnitine clearance and Na+‐dependent muscle carnitine accretion, likely via stimulating Na+/K+ ATPase pump activity. We hypothesized that the ingestion of high‐dose caffeine, also known to stimulate Na+/K+ ATPase activity, would stimulate plasma carnitine clearance during hypercarnitinemia in humans. In a randomized placebo‐controlled study, six healthy young adults (aged 24 ± 5 years, height 175 ± 8 cm, and weight 70 ± 13 kg) underwent three 5‐h laboratory visits involving the primed continuous intravenous infusion of l‐carnitine (CARN and CARN + CAFF) or saline (CAFF) in parallel with ingestion of caffeine (CARN + CAFF and CAFF) or placebo (CARN) at 0, 2, 3, and 4 h. Regular blood samples were collected to determine concentrations of blood Na+ and K+, and plasma carnitine and caffeine, concentrations. Caffeine ingestion (i.e., CAFF and CARN + CAFF conditions) and l‐carnitine infusion (i.e., CARN and CARN + CAFF) elevated steady‐state plasma caffeine (to ~7 μg·mL−1) and carnitine (to ~400 μmol·L−1) concentrations, respectively, throughout the 5 h infusions. Plasma carnitine concentration was ~15% lower in CARN + CAFF compared with CARN during the final 90 min of the infusion (at 210 min, 356 ± 96 vs. 412 ± 94 μmol·L−1; p = 0.0080: at 240 min, 350 ± 91 vs. 406 ± 102 μmol·L−1; p = 0.0079: and at 300 min, 357 ± 91 vs. 413 ± 110 μmol·L−1; p = 0.0073, respectively). Blood Na+ concentrations were greater in CAFF and CARN + CAFF compared with CARN. Ingestion of high‐dose caffeine stimulates plasma carnitine clearance during hypercarnitinemia, likely via increased Na+/K+ ATPase activity. Carnitine co‐ingestion with caffeine may represent a novel muscle carnitine loading strategy in humans, and therefore manipulate skeletal muscle fuel metabolism and improve exercise performance.

Relative contribution of neuromuscular activation, muscle size, and muscle quality to maximum strength output of the thigh muscles in young individuals

Abstract: This study aimed to investigate the relationship between maximal muscle strength and neuromuscular activation, muscle size, and quality of quadriceps (QF) and hamstring muscles (HM). The study included 24 young men and women. The neuromuscular activation parameter was recorded using a single‐channel surface electromyography (EMG) with the root mean square (RMS) during maximal isometric knee extension and flexion from four muscles: rectus femoris and vastus lateralis for QF; biceps femoris and semitendinosus for HM. In addition, the peak torque was measured during the same session. B‐mode ultrasonographic transverse images were obtained from the anterior, lateral, and posterior thighs. Furthermore, we calculated the muscle thickness (MT) and echo intensity (EI) of the four muscles as indicators of muscle size and quality. The averaged MT, EI, and absolute RMS of QF were calculated by averaging the values of the rectus femoris and vastus lateralis, and that of HM was calculated by averaging the values of the biceps femoris and semitendinosus. The knee extension peak torque was correlated with EI (r = −0.61, P < 0.01) and RMS (r = 0.53, P < 0.01) in the QF. In contrast, the knee flexion peak torque was correlated with RMS (r = 0.53, P < 0.05) but not with MT and EI in HM. In addition, EI and RMS in QF, and RMS in HM were selected as the major determinants of muscle strength in the stepwise regression analysis. These results suggest that muscle strength is moderately associated with different factors related to the thigh muscles in young individuals.

Unraveling pathophysiologic mechanisms contributing to symptoms in patients with post‐acute sequelae of COVID‐19 (PASC): A retrospective study

Abstract: Patients with post‐acute sequelae of COVID‐19 (PASC) present with a decrease in physical fitness. The aim of this paper is to reveal the relations between the remaining symptoms, blood volume distribution, exercise tolerance, static and dynamic lung volumes, and overall functioning. Patients with PASC were retrospectively studied. Pulmonary function tests (PFT), 6‐minute walk test (6MWT), and cardiopulmonary exercise test were performed. Chest CT was taken and quantified. Patients were divided into two groups: minor functional limitations (MFL) and severe functional limitations (SFL) based on the completed Post‐COVID‐19 Functional Status scale (PCFS). Twenty one patients (3 M; 18 FM), mean age 44 (IQR 21) were studied. Eighteen completed the PCFS (8 MFL; 10 SFL). VO2max was suboptimal in both groups (not significant). 6MWT was significantly higher in MFL‐group (p = 0.043). Subjects with SFL, had significant lower TLC (p = 0.029). The MFL‐group had more air trapping (p = 0.036). Throughout the sample, air trapping correlated significantly with residual volume (RV) in L (p < 0.001). An increase in air trapping was related to an increase in BV5 (p < 0.001). Mean BV5 was 65% (IQR 5%). BV5% in patients with PASC was higher than in patients with acute COVID‐19 infection. This increase in BV5% in patients with PASC is thought to be driven by the air trapping in the lobes. This study reveals that symptoms are more driven by occlusion of the small airways. Patients with more physical complaints have significantly lower TLC. All subjects encounter physical limitations as indicated by suboptimal VO2max. Treatment should focus on opening or re‐opening of small airways by recruiting alveoli.

Exercise training modifies xenometabolites in gut and circulation of lean and obese adults

Abstract: Regular, moderate exercise modifies the gut microbiome and contributes to human metabolic and immune health. The microbiome may exert influence on host physiology through the microbial production and modification of metabolites (xenometabolites); however, this has not been extensively explored. We hypothesized that 6 weeks of supervised, aerobic exercise 3×/week (60%–75% heart rate reserve [HRR], 30–60 min) in previously sedentary, lean (n = 14) and obese (n = 10) adults would modify both the fecal and serum xenometabolome. Serum and fecal samples were collected pre‐ and post‐6 week intervention and analyzed by liquid chromatography/tandem mass spectrometry (LC–MS/MS). Linear mixed models (LMMs) identified multiple fecal and serum xenometabolites responsive to exercise training. Further cluster and pathway analysis revealed that the most prominent xenometabolic shifts occurred within aromatic amino acid (ArAA) metabolic pathways. Fecal and serum ArAA derivatives correlated with body composition (lean mass), markers of insulin sensitivity (insulin, HOMA‐IR) and cardiorespiratory fitness ( V̇O2max$$ \dot{\mathrm{V}}{\mathrm{O}}_{2\max } $$ ), both at baseline and in response to exercise training. Two serum aromatic microbial‐derived amino acid metabolites that were upregulated following the exercise intervention, indole‐3‐lactic acid (ILA: fold change: 1.2, FDR p < 0.05) and 4‐hydroxyphenyllactic acid (4‐HPLA: fold change: 1.3, FDR p < 0.05), share metabolic pathways within the microbiota and were associated with body composition and markers of insulin sensitivity at baseline and in response to training. These data provide evidence of physiologically relevant shifts in microbial metabolism that occur in response to exercise training, and reinforce the view that host metabolic health influences gut microbiota population and function. Future studies should consider the microbiome and xenometabolome when investigating the health benefits of exercise.

Cardiomyocyte ZKSCAN3 regulates remodeling following pressure‐overload

Abstract: Autophagy is important for protein and organelle quality control. Growing evidence demonstrates that autophagy is tightly controlled by transcriptional mechanisms, including repression by zinc finger containing KRAB and SCAN domains 3 (ZKSCAN3). We hypothesize that cardiomyocyte‐specific ZKSCAN3 knockout (Z3K) disrupts autophagy activation and repression balance and exacerbates cardiac pressure‐overload‐induced remodeling following transverse aortic constriction (TAC). Indeed, Z3K mice had an enhanced mortality compared to control (Con) mice following TAC. Z3K‐TAC mice that survived exhibited a lower body weight compared to Z3K‐Sham. Although both Con and Z3K mice exhibited cardiac hypertrophy after TAC, Z3K mice exhibited TAC‐induced increase of left ventricular posterior wall thickness at end diastole (LVPWd). Conversely, Con‐TAC mice exhibited decreases in PWT%, fractional shortening (FS%), and ejection fraction (EF%). Autophagy genes (Tfeb, Lc3b, and Ctsd) were decreased by the loss of ZKSCAN3. TAC suppressed Zkscan3, Tfeb, Lc3b, and Ctsd in Con mice, but not in Z3K. The Myh6/Myh7 ratio, which is related to cardiac remodeling, was decreased by the loss of ZKSCAN3. Although Ppargc1a mRNA and citrate synthase activities were decreased by TAC in both genotypes, mitochondrial electron transport chain activity did not change. Bi‐variant analyses show that while in Con‐Sham, the levels of autophagy and cardiac remodeling mRNAs form a strong correlation network, such was disrupted in Con‐TAC, Z3K‐Sham, and Z3K‐TAC. Ppargc1a also forms different links in Con‐sham, Con‐TAC, Z3K‐Sham, and Z3K‐TAC. We conclude that ZKSCAN3 in cardiomyocytes reprograms autophagy and cardiac remodeling gene transcription, and their relationships with mitochondrial activities in response to TAC‐induced pressure overload.

Muscle sympathetic nerve activity during pregnancy: A systematic review and meta‐analysis

Abstract: We conducted a systematic review and meta‐analysis to quantify the impact of healthy and complex pregnancy on muscle sympathetic nerve activity (MSNA) at rest, and in response to stress. Structured searches of electronic databases were performed until February 23, 2022. All study designs (except reviews) were included: population (pregnant individuals); exposures (healthy and complicated pregnancy with direct measures of MSNA); comparator (individuals who were not pregnant, or with uncomplicated pregnancy); and outcomes (MSNA, BP, and heart rate). Twenty‐seven studies (N = 807) were included. MSNA burst frequency was higher in pregnancy (n = 201) versus non‐pregnant controls (n = 194) (Mean Differences [MD], MD: 10.6 bursts/min; 95% CI: 7.2, 14.0; I2 = 72%). Accounting for the normative increase in heart rate with gestation, burst incidence was also higher during pregnancy (Pregnant N = 189, non‐pregnant N = 173; MD: 11 bpm; 95% CI: 8, 13 bpm; I2 = 47%; p < 0.0001). Meta‐regression analyses confirmed that although sympathetic burst frequency and incidence are augmented during pregnancy, this was not significantly associated with gestational age. Compared to uncomplicated pregnancy, individuals with obesity, obstructive sleep apnea, and gestational hypertension exhibited sympathetic hyperactivity, while individuals with gestational diabetes mellitus or preeclampsia did not. Uncomplicated pregnancies exhibited a lesser response to head‐up tilt, but an exaggerated sympathetic responsiveness to cold pressor stress compared to non‐pregnant individuals. MSNA is higher in pregnant individuals and further increased with some, but not all pregnancy complications. PROSPERO registration number: CRD42022311590.

Effect of after‐school physical activity on body composition in primary school children: The Slovak “PAD” project

Abstract: Physical activity is associated with many physical and mental health benefits. This study aimed to investigate the effect of a 24‐month after‐school physical activity intervention on body composition in normal‐weight children. Participating students (6–7 years of age at baseline) were divided by reason of their parental preference to intervention and control groups. Children in the intervention group (n = 20; 10 boys and 10 girls) followed an aerobic training program (two 60‐min sessions per week), whereas children in the control group (n = 20; 10 boys and 10 girls) participated in the usual practice. Body composition characteristics were repeatedly measured by means of bioelectrical impedance method. At 2 years, finally, intervention boys had a smaller rise in BMI (mean difference, MD: −0.97 kg/m2, p < 0.05), BMI z‐score (−0.44, p < 0.09), body fat % (BF%) (−6.47%, p < 0.01), and fat mass index (FMI) (−1.32 kg/m2, p < 0.001) than controls. In girls, however, the intervention program induced no significant differences (p > 0.9) in the measured variables compared to controls at the final follow‐up (MD: −0.04 kg/m2 for BMI and −0.01 for BMI z‐score). Changes in BF% and FMI in a positive direction occurred at 18 months (MD: −3.38%, p < 0.05 and −0.99 kg/m2, p < 0.01, respectively), but did not persist over time (p > 0.07). In addition, no significant changes (p > 0.07) in the fat‐free mass index were associated with the physical activity intervention in either boys or girls. In conclusion, compared to the controls, a long‐term physical activity intervention in boys was associated with a significantly smaller rise in BMI and improvement of body composition by reducing both BF % and FMI. In girls, however, this intervention did not result in any statistically significant changes in body composition variables.

Diabetes mellitus differently affects electrical membrane properties of vagal afferent neurons of rats

Abstract: To study whether diabetes mellitus (DM) would cause electrophysiological alterations in nodose ganglion (NG) neurons, we used patch clamp and intracellular recording for voltage and current clamp configuration, respectively, on cell bodies of NG from rats with DM. Intracellular microelectrodes recording, according to the waveform of the first derivative of the action potential, revealed three neuronal groups (A0, Ainf, and Cinf), which were differently affected. Diabetes only depolarized the resting potential of A0 (from −55 to −44 mV) and Cinf (from −49 to −45 mV) somas. In Ainf neurons, diabetes increased action potential and the after‐hyperpolarization durations (from 1.9 and 18 to 2.3 and 32 ms, respectively) and reduced dV/dtdesc (from −63 to ‐52 V s−1). Diabetes reduced the action potential amplitude while increasing the after‐hyperpolarization amplitude of Cinf neurons (from 83 and −14 mV to 75 and −16 mV, respectively). Using whole cell patch clamp recording, we observed that diabetes produced an increase in peak amplitude of sodium current density (from −68 to −176 pA pF−1) and displacement of steady‐state inactivation to more negative values of transmembrane potential only in a group of neurons from diabetic animals (DB2). In the other group (DB1), diabetes did not change this parameter (−58 pA pF−1). This change in sodium current did not cause an increase in membrane excitability, probably explainable by the alterations in sodium current kinetics, which are also induced by diabetes. Our data demonstrate that diabetes differently affects membrane properties of different nodose neuron subpopulations, which likely have pathophysiological implications for diabetes mellitus.