bluuebunny/crossref_metadata_2025 huggingface dataset:
Rutin ameliorates high cholesterol diet induced alteration in lipid profile Interleukin‐3 (IL3) and Interleukin6 (IL6) and liver inflammation
Authors: Alhosaini Khaled , AlSharari Shakir , Hafez Mohammed M.
Published: November 2023

Non alcoholic fatty liver disease (NAFLD) has become a very common metabolic disorder. It refers to a group of conditions where excess fats are deposited in hepatic cells. High cholesterol diet (HCD) causes dyslipidemic syndrome and hyperlipidemia. Furthermore, hypercholesterolemia induceshepatic oxidative stress and inflammation and activation of inflammatory mediators and suppression of the anti‐inflammatory genes. The natural flavonoid (Rutin) possesses an antioxidant activity with protective potential. Therefore, we designed this study with the aim to investigate the potential ameliorative potential of rutin against HCD‐induced alteration of lipid profile and hepatic interleukins; Interleukin‐3 (IL3) and Interleukin6 (IL6). Male Wistar rats were allocated into four groups as follow (1) control (Rat chow), (2) Rutin (0.2% in rat chow), (3) HCD (1% cholesterol and 0.5% cholic acid in rat chow) and (4) rutin (0.2%) + HCD. Animals received freshly prepared experimental diets for 6 consecutive weeks. In plasma, levels triglyceride (TG), total cholesterol (TC), low‐density lipoprotein (LDL) and high‐density lipoprotein (HDL)were estimated. In hepatic tissue, gene expressions of Interleukin‐3 (IL3) and Interleukin6 (IL6) were determined using real time PCR. HCD significantly increased plasma levels of TG, TC and LDL by 48%, 89% and 67% respectively and significantly decreased the HDL levels by 17% compared to control group. Rutin supplementation in combination with HCD, significantly decreased TG and LDL levels and insignificant decrease plasma levels of TC and HDL compared to control group. Results also showed a significant increase by 5.8 fold in the expression of IL‐6 and significant decrease by 3 folds in IL‐3 expression in rats fed with HCD compared to control group. Interestingly, administration of rutin in combination with HCD corrected the alteration in the expression of these genes to their values in the control group. In conclusion: Combination of the natural occurring antioxidant, Rutin, with HCD protects against HCD associated hepatic inflammation and the altered lipid profile.

Support or Funding Information

king saud university
TRPM8 Mediates the Anti‐inflammatory Effects of Eucalyptol in a Mouse Model of LPS‐Induced Pulmonary Inflammation
Authors: Caceres Ana Isabel , Liu Boyi , Jabba Sairam V. , Morris John B. , Jordt Sven‐Eric
Published: November 2023

Eucalyptol (1,8‐cineol), the major ingredient in the essential oil of the eucalyptus leaves and other medicinal plants, has long been known for its anti‐inflammatory properties. Eucalyptol interacts with Transient Receptor Potential (TRP) ion channels among other targets, but it is unclear which of these mediates its anti‐inflammatory effects. In this work, the effects of eucalyptol were compared in WT and TRPM8‐deficient mice in a model of pulmonary inflammation following intranasal administration of lipopolysaccharide (LPS). LPS, a component of Gram‐negative bacterial cell walls, plays a major role in acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). 24 hours after LPS intranasal instillation lungs and bronchoalvelolar lavage (BAL) were collected and analyzed for leukocyte infiltration, enzyme activities and cytokine/chemokine levels by qPCR, Luminex and cell counting among other techniques.

Our results show that eucalyptol treatment diminished leukocyte infiltration, myeloperoxidase activity and production of TNF‐α, IL‐1β, IFN‐γ and IL‐6. Genetic deletion of TRPM8 abolishes the anti‐inflammatory effects of eucalyptol in the LPS model. Eucalyptol is at least 30‐fold more potent on human TRPM8 than on mouse TRPM8, and a metabolite of eucalyptol, 2‐hydroxy‐1,8‐cineol, also activates human TRPM8. This shows that among the pharmacological targets of eucalyptol, TRPM8 is essential for its anti‐inflammatory effects in mice, likely mediated by peripheral sensory neurons. The much higher sensitivity of human TRPM8 to eucalyptol compared to the rodent orthologues explains the higher potency of eucalyptol in humans. Metabolites of eucalyptol likely contribute to the anti‐inflammatory effects. The development of more potent and selective TRPM8 agonists may yield novel anti‐inflammatory therapeutics.

Support or Funding Information

‐ NIH grants R01HL105635 and R01HL105635‐S1 (to S. E. J. and J. B. M.)

‐ P50DA036151 project #1 (to S.E.J.).
PHOSPHODIESTERASE (PDE) INHIBITORS AND THEIR INTERACTION WITH NITRIC OXIDE (NO) MODULATORS IN ADJUVANT‐INDUCED RHEUMATOID ARTHRITIS
Authors: Pal Rishi , Tiwari Prafulla Chandra , Nath Rajendra , Babu Suresh , Pant Kamlesh Kumar
Published: November 2023

Phosphodiesterase (PDE) inhibitors are known to increase cAMP/cGMP in the cellular compartment and leads to signal transduction process in the various cell system. Here we focused on the anti‐inflammatory & immunomodulatory protective role of some PDE inhibitors theophylline, pentoxifylline & rolipram and their interactions with nitric oxide (NO) modulators in complete Freund's adjuvant induced rheumatoid arthritis in rats. Wistar rats (200–300 gm, n=6/group) of either sex were used in the study. On day ‘0’ rats were injected with 0.2 ml of complete Freund's adjuvant (CFA) in sub‐planter region of right hind paw along with 0.1 ml of squalene to develop RA while controls received only vehicle. PDE inhibitors drug treatment alone and in combination with NO modulators was given from day ‘14’ to ‘28’. Arthritic parameters a) arthritis index, b) ankle diameter c) paw volume and their body weight were noted to evaluate progression of RA on day 0, 7, 14, 21 and 28. On day ‘28’ rats were sacrificed and their blood and paws were collected for TNF‐α and IL‐10 cytokine estimation, pathological examination and NF‐kB expression. Data obtained was analysed using two‐way ANOVA followed by Newman‐Keul's posthoc test and p<0.05 was considered for significance. It was found that CFA significantly increased arthritis‐index, paw volume, ankle diameter and serum TNF‐α and NF‐kB levels while body weight and serum IL‐10 levels was significantly decreased (P<0.05). These CFA‐induced changes were significantly reversed by theophylline (10 & 20 mg/kg), P<0.0001); pentoxifylline (5 & 10 mg/kg), P<0.002); rolipram (1 & 2 mg/kg), P<0.05) alone and in combination with L‐arginine (100 mg/kg) and or L‐NAME (10 mg/kg) in dose dependent manner (p<0.005) in all parameters). The maximum protective effects was observed in theophylline > pentoxifylline > rolipram with L‐NAME 10 mg/kg combination treatment group (p<0.001). The data obtained was substantiated histopathologically. Results of this study are suggestive of the involvement & interaction of NO with different PDE inhibitors may have protective role in adjuvant induced RA with may be mediated through cytokine signalling pathways.

Support or Funding Information

The research work financially supported by research cell, King George's Medical University, UP, Lucknow in the form of Intramural research grant awarded to First author Dr. Rishi Pal.
Glutathione Increases in Response to a Glucose Challenge in Diabetic Rat Hearts
Authors: Thorwald Max Andrew , Rodriguez Gema J , Rodriguez Marco Antonio , Nakano Daisuke , Nishiyama Akira , Ortiz Rudy M
Published: November 2023

Diabetic hearts are in great risk of increased oxidant production from sustained elevations of glucose through activation of several pathways. One of the major sources of oxidant production within the cell are mitochondria through substrate metabolism. Oxidation of mitochondrial enzymes may impair their function resulting in accumulation of intermediates and reverse electron transfer which contributes to mitochondrial dysfunction. Glutathione (GSH) is the most abundant antioxidant in the body which with the aid of peroxidases is able to reduce cellular oxidants through electron donation. Transcription factor nuclear factor erythriod‐2‐related factor 2 (Nrf2) induces expression of antioxidant genes including those responsible for GSH synthesis. Nrf2 translocation serves as a defensive mechanism when oxidant production increases in the cytosol. We hypothesized that an elevation in plasma glucose facilitates Nrf2 nuclear translocation and subsequent expression of Nrf2‐associated genes. To address this hypothesis, an oral glucose tolerance test (oGTT) was performed in four groups (n = 6–8/group) of rats: 1) LETO (lean strain‐control), 2) vehicle‐treated, IR obese OLETF, 3) OLETF + angiotensin receptor blocker (ARB; 10 mg olmesartan/kg/d), and 4) OLETF + angiotensin receptor blocker (ARB‐r). ARB normalized systolic blood pressure to control levels, while ARB‐r matched OLETF at 155±2.36 mmHg. Baseline GSH levels were maintained over 360 min post‐glucose administration in LETO, but GSH levels increased 125% in OLETF. While ARB had no detectable effect on GSH levels, GSH levels increased 52% in ARB‐r compared to LETO. Removal of ARB treatment may be more beneficial than long term ARB treatment. However, our data suggest that acute increases in glucose can increase GSH levels in the early phases of diabetic cardiomyopathy liking to protect against the potential for increased oxidant production.
Neutrophil‐derived 20‐HETE Is a Major Determinant of Coronary Collateral Growth
Authors: Joseph Gregory , Hutcheson Brenda , Hunter Ian , Soler Amanda , Gotlinger Katie , Falck John , Schwartzman Michal L , Rocic Petra
Published: November 2023

Coronary collateral growth (CCG) is an adaptive response to transient, repetitive coronary artery occlusion in normal animals but is markedly impaired in metabolic syndrome. Recent studies in our laboratory have shown that in metabolic syndrome rats (JCR:LA‐cp, JCR) in addition to endothelial cell (EC) dysfunction, neutrophil survival is enhanced, leading to neutrophil accumulation which contributes to compromised CCG in JCR rats. 20‐hydroxyeicosatetraenoic acid (20‐HETE) plays an important role in vascular function. However, the role of 20‐HETE in the regulation of CCG is unknown. We investigated the hypothesis that elevated neutrophil‐derived 20‐HETE promotes EC dysfunction and apoptosis and impairs CCG in metabolic syndrome. In our study, coronary blood flow in Sprague‐Dawley (SD) or JCR rats was measured in the collateral (CZ) and normal zone (NZ) of the heart after 9 days of transient, repetitive LAD occlusion (repetitive ischemia, RI) using microspheres. Collateral growth in the JCR rats was impaired compared to SD rats (CZ/NZ flow ratio was 0.11±0.02 in JCR vs. 0.84±0.02 in SD). Administration of 20‐HETE antagonists, either 20‐SOLA or 20‐HEDGE, fully restored collateral‐dependent blood flow (0.85±0.01 in JCR+20‐SOLA, 0.84±0.01 in JCR+20‐HEDGE), while neutrophil depletion, via neutrophil‐blocking antibodies, partially restored collateral‐dependent blood flow (0.57±0.02) in JCR rats. Neutrophil accumulation was ~3 times higher and persisted for the duration of the RI protocol in the CZ of JCR vs. SD rats, where their infiltration was low and transient. This RI‐induced, elevated neutrophil infiltration in the CZ of JCR rats correlated with a >4‐fold increase in CYP4F (the neutrophil 20‐HETE CYP isoform) expression and a 4‐fold increase in 20‐HETE levels vs. SD rats, which was abolished by administration of neutrophil‐blocking antibodies to JCR rats. Furthermore, coronary microvessels of JCR rats displayed reduced endothelium‐dependent vasodilation after 9 days of RI (20%±1.2% JCR vs. 40%±2.3% SD), which was reversed with neutrophil‐blocking antibodies (37%±2.5% JCR+anti‐CD11b/CD18/CD44). eNOS phosphorylation (p1179) and NO production were likewise decreased (~50%) in the CZ of JCR rays after 9 days of RI, which was reversed after administration of neutrophil‐blocking antibodies. Lastly, RI‐induced EC apoptosis in JCR but not in SD rats (TUNEL). EC viability in JCR rats was almost completely restored by treatment with neutrophil‐blocking antibodies. Taken together, these results indicate that neutrophil‐derived 20‐HETE is a major contributor to impaired EC viability and function, and thus to impaired CCG in metabolic syndrome.

Support or Funding Information

Support: NIH R01 HL093052.
How Important Is SERCA In Neuronal Calcium Homeostasis?
Authors: Shanks Julia , Paterson David
Published: November 2023

Maintaining intracellular calcium homeostasis is critical for excitation‐contraction coupling in the heart, and is essentially dependent on primary and secondary active transport systems. However, upregulating sarcoendoplasmic reticulum calcium ATPase (SERCA) in cardiomyocytes from patients with heart failure has recently proved inconclusive (CUPID2 trial). Neuronal calcium impairment is also seen in cardiovascular diseases resulting in heightened sympathetic responsiveness that further exacerbate the disease phenotype. We tested whether overexpression of SERCA in cardiac sympathetic neurons could modulate intracellular calcium transients. Isolated neurons were incubated with either Ad‐MCherry‐SERCA2A or Ad‐MCherry‐Empty (control) for 24 hours immediately post isolation. Calcium imaging using fura‐2AM (2.5μM) was carried out three days post transfection. SERCA2A transgene neurons showed greater depolarisation induced intracellular calcium transients, compared to control (Δ 380/340 ratio (au): SERCA 0.75±0.03, n=68; Empty 0.64±0.03, n=57. P<0.01). SERCA2A expressing neurons also showed greater SR calcium storage compared to control, as demonstrated by significantly higher calcium responses to caffeine (10mM: Δ 380/340 ratio (au): SERCA 0.15±0.01, n=45; Empty 0.03±0.01, n=35. P<0.0001) and thapsigargin stimulation (1μM: Δ 380/340 ratio (au): SERCA 0.12±0.02, n=42; Empty 0.03±0.01, n=33. P<0.0001). Additionally mitochondrial calcium uncoupling with FCCP (1μM) resulted in a greater intracellular calcium change in the SERCA2A transgenic neurons (Δ 380/340 ratio (au): SERCA 0.13±0.01, n=22; Empty 0.05±0.01, n=22. P<0.0001), indicating that altering neuronal calcium handling with upregulation of SERCA has also effected mitochondrial calcium handling and potentially metabolic function in these neurons. Within this study we demonstrated that a SERCA2A gene construct can be transfected in isolated sympathetic neurons from the stellate ganglia. Moreover, this transduction caused an increase in intracellular calcium transients that would be expected to facilitate neurotransmission and be potentially deleterious for cardiac function.

Support or Funding Information

British Heart Foundation
Impact Of Hypoxia In The Expression And Regulation Of The TASK‐1 Potassium Channel In Cardiac Myocytes
Authors: Kang Chen , Hu Keli
Published: November 2023

TASK‐1 is a member of the two‐pore domain potassium channel family and strongly expressed in the heart. However, little is known about the roles and regulation of TASK‐1 channels in cardiac hypoxia. The present study was designed to investigate whether hypoxia modulated the expression and function of TASK‐1 and whether protein kinase C (PKC) was critical in hypoxia‐mediated processes in adult rat cardiac myocytes. The effects of acute hypoxia on the TASK‐1 channel function were chemically induced by sodium cyanide (NaCN). Activity of TASK‐1 in cardiomyocytes was investigated by the whole‐cell patch‐clamp technique. The current flowing through TASK‐1 channels in cardiac myocytes was quantified by analyzing the effects of extracellular pH or the selective TASK‐1 inhibitor methanandamide on the steady‐state current‐voltage relation. The voltage protocol, which included voltage ramps from +40 to −100 mV preceded by a test pulse from 0 to +40 mV, was designed to inactivate voltage‐dependent ion channels. In addition, a blocker cocktail, that had no effect on TASK‐1 channels expressed in HEK293T cells, was used to eliminate a number of ion channels in cardiac myocytes. Our data show that exposure of cardiac myocytes to NaCN for 5 minutes reduced the steady‐state outward current at +40 mV, an effect that was abolished by pretreatment with methanandamide. Either methanandamide or extracellular acidification (from pH 7.4 to 6.4) decreased the outward current, identifying TASK‐1 as the channel mediating this effect. Interestingly, the NaCN‐induced reduction in the outward current was largely prevented by pretreatment with the PKC selective inhibitor bisindolylmaleimide. To assess the expression and protective role of TASK‐1 in hypoxia, the prolonged hypoxia (4 hours) was employed by incubating cells in an airtight chamber in which O2 was replaced by N2 with glucose‐free Tyrode's solution. While 4 hours of hypoxia caused a significant reduction in the viability of cardiac myocytes, the inhibition of cardiac TASK‐1 channels by methanandamide further increased hypoxia‐induced cell death. Western blot analysis showed that 4 hours of hypoxia led to a decrease in the protein expression of TASK‐1 in cardiac myocytes. In summary, we provided the evidence that TASK‐1 channels in cardiomyocytes is regulated by hypoxia‐induced PKC signaling in cardiomyocytes. Our data are also supportive for a possible role of TASK‐1 channels in the cardioprotection against hypoxia.
Combining Mass Spectrometry Cross‐Linking, Molecular Docking and NMR Titration to Dissect ZapA‐FtsZ Protein Interaction
Authors: Nogueira Maria Luiza Caldas , Sforça Mauricio Luis , Leme Adriana Franco Paes , Pauletti Bianca Alves , de Oliveira Paulo Sergio Lopes , Honorato Rodrigo Vargas , King Glenn F , Filho Frederico José Gueiros , de Mattos Zeri Ana Carolina
Published: November 2023

The cell division is a key event in a bacteria cycle of life and in most rod shaped bacteria. This cell divion involves a septum formation. FtsZ and other proteins forms a macromolecular complex, named Divisome, which redirect the cell wall and provide a constrictive force required to divide the bacterial cell in two daughter cells. FtsZ is a tubulin homologue that polymerizes as a ring structure shaped, the Z ring. The Divisome is composed by: the Z ring, modulators proteins, proteins that anchor the Z ring to the cell membrane and synthesize cell wall (peptidoglycan). Modulators proteins are essential for the cell division occurs at the right place and time, because they act stimulating or preventing the Z ring formation. Defects on modular proteins could cause defects in bacterial cell division such as filamentation and mini‐cell formation. The Z ring associated protein A (ZapA) is a positive modulator that assists the bundling of FtsZ polymers by promote lateral association of FtsZ filaments. Despite the interaction between FtsZ‐ZapA was described many years ago, the molecular mechanisms details involved into this process is barely understood. Here we combined the information obtained from FtsZ‐ZapA NMR titration and Mass Spectrometry cross‐linking to modelling the FtsZ and ZapA protein complex. The docking model may give us a clue of the main amino acids residues involved on that interaction and how thus it occurs.

Support or Funding Information

São Paulo Research Foundation (FAPESP); State University of Campinas (UNICAMP) and Brazilian Center for Research in Energy and Materials/Brazilian Biosciences National Laboratory ( CNPEM/LNBio )
Effects of Sleep Apnea in Chronic Kidney Disease
Authors: Pace Carolyn , Riggs Jennifer , Danielson Leslie , Fischer Edgar , Howard Tamara , Kanagy Nancy
Published: November 2023

Patients with chronic kidney disease (CKD) have increased incidence of sleep apnea compared to those without renal disease. We hypothesized that the hypoxia of sleep apnea accelerates declines in renal function and so might contribute to this association. In our model of CKD (0.4% adenine diet for 3 to 5 weeks), rats developed crystalline nephropathy and elevated blood urea nitrogen (BUN) values (> 140 mg/dl). After return to a normal rat diet, BUN decreased to a stable but still elevated level of 50–70 mg/dl simulating CKD (normal rat BUN = 20 mg/dl). 24‐hour urine volumes were also significantly elevated (CKD = 38±3 vs Control = 9.4±0.5 ml). Urinary protein excretion increased in the adenine diet rats but not in the control diet rats. Rats were then exposed during their sleep period to either intermittent hypoxia (IH) to simulate sleep apnea or sham conditions for six weeks. At the end of the IH or sham exposures, BUN levels were greater in CKD rats exposed to IH compared to Sham conditions (Sham = 78.3±3.7 vs. IH = 91.5±3.1 mg/dl) but were not different in non‐CKD groups. Creatinine clearance was lower in rats exposed to IH for both CKD rats (Sham = 1.0±0.2 vs. IH = 0.81±0.1 ml/min) and in control rats (Sham = 4.2±0.7 vs. IH = 1.6±0.1 ml/min) suggesting IH impairs glomerular function. Histological analysis of kidney sections revealed fibrosis and infiltrates in the regions of the proximal and distal tubules with effaced proximal tubules and decreased total number of tubules in all adenine fed rats. This damage persisted 8 weeks after cessation of the diet, independent of sham or IH exposure. Therefore, an adenine diet causes sustained CKD even after the diet is discontinued that results in polyuria and proteinuria. Exposure to IH further compromises renal function suggesting treating sleep apnea in CKD patients should help to preserve renal filtration function.

Support or Funding Information

Dialysis Clinic, Inc. (DCI)
Does Exposure to Nanoparticles Alter the Resistance of Bacteria to Antibiotics and Heavy Metals?
Authors: Paek Ashley , Smith Virginia
Published: November 2023

Titanium dioxide (TiO2) nanoparticles demonstrate the properties of high refractive index and brightness and therefore, are regularly used as reflective optical coats or as whitening pigments. In addition, TiO2 is also used extensively in cosmetics, skin care and sunscreen products due to its transparent appearance under visible light. As the applications for nanoparticles (NP) increase, it is becoming more urgent to understand the levels and mechanisms of their toxicity. Previous work in our lab, and others, has determined that while high levels of NP are toxic to bacteria, low levels of NP actually enhance bacterial growth rates (hormesis). Recent studies have shown that the expression of heavy‐metal and antibiotic efflux pumps is upregulated in response to exposure to nanoparticles. In order to see if such upregulation was responsible for the hormesis observed in our lab, we developed and performed disk diffusion assays on NP‐treated bacterial cultures. Paper disks that had been pre‐treated with different classes of antibiotics or types of metal ions were placed on agar plates containing cultures of E.coli that had either been exposed to nanoparticles or had not. The plates were then incubated overnight to allow the contents of the disk to diffuse into the surrounding agar. Preliminary results indicate that exposure to nanoparticles confers minimal, if any, resistance to ampicillin or lead, nickel, or zinc metal ions. Ongoing tests are investigating other antibiotics, metal ions, and levels of nanoparticle exposure.

Support or Funding Information

Defense Threat Research Agency
Initial Characterization of the Pig Skin Bacteriome and its Effect on in vitro Models of Wound Healing
Authors: McIntyre Matthew K. , Baker Bryan , Peacock Trent J. , Burmeister David M
Published: November 2023

Improved combat casualty care has increased survivability of traumatic wartime injuries, resulting in an emphasison longer‐term outcome such as wound healing. Recent advances in understanding the roles and composition of the humanskin microbiome have revealed a delicate interplay between resident microbes and combat wound healing. While pigs are the gold standard model for wound healing due to structural and healing similarities to human skin, the porcineskin microbiome has not been adequately elucidated. Moreover, no model organism has been established to examine the effects of novel pharmaceuticals on the skin microbiome in vivo. In the current study, we performed 16S rRNA gene sequencing on different anatomical locations from porcine skin, and utilized in vitro techniques to examine the paracrine effects of proteins released from these bacteria on human keratinocytes (NHEK) and fibroblasts (HDF). We found that the pig skin bacterial diversity varies in quantity and composition between subjects and anatomical locations. Moreover, t‐tests revealed significant effects on HDF and NHEK migration that was dependent on anatomical location. ELISA showed that, when cultured with NHEKs, bacterial releasate from certain anatomical locations reduced the amount of Vascular Endothelial Growth Factor present in the media and also induced NHEK apoptosis as determined via caspase‐3immunostaining. The results described herein further expand the knowledge of the cutaneous bacteriome of the healthy pig and suggest that naturally occurring bacterial flora may affect wound healing. This is particularly intriguing considering the amount of resources utilized for combat casualty wound care (i.e. healing, scarring, antibiotic resistance). Ultimately, if similarities between human and pig skin extend to the level of microbial diversity, the pig might be considered an ideal model for further investigations into the naturally occurring flora of the skin and its impact on wound healing process.
Markers of Muscle Protein Synthesis and Breakdown in Fast‐Twitch Skeletal Muscle of Rodents Aged 3 to 24 Months
Authors: Mumford Petey W. , Mobley C. Brooks , Kephart Wesley C. , Haun Cody T. , Holland A. Maleah , Osburn Shelby C. , Beck Darren T. , Martin Jeffery M. , Young Kaelin C. , Kavazis Andreas N. , Lowery Ryan P. , Wilson Jake M. , Roberts Michael D.
Published: November 2023

OBJECTIVE

Muscle protein synthesis (MPS) and muscle protein degradation (MPD) are well known modulators for muscle mass maintenance. Given that fast‐twitch skeletal muscle atrophies with aging, we sought to examine the effects of aging on MPS and MPD markers in the plantaris muscle of rodents ranging from 3 to 24 months.

MEHTODS

Male Fischer 344 rats (300–600g) were successfully aged to 3, 6, 12, 18 and 24 months, were injected with puromycin (0.02 mg/g body mass), euthanized, plantaris muscle was extracted and wet skeletal muscle weights were obtained. Muscle tissue was then processed for biochemical processing. MPS was assessed via the SUNsET method, and MPD was assessed using whole lane densitometry via ubiquitin immunoblotting.

RESULTS

Relative plantaris masses revealed significant between‐group differences throughout the lifespan (p<0.001); specifically, masses were 18% greater in 3 versus 18 month old rats (p<0.001) and 23% greater in 3 versus 24 month old rats (p<0.001). Interestingly, MPS levels did not differ between 3 month old rats versus all other age groups. However, MPS levels were 2.42‐fold greater in 18 month old versus 24 month old rats (p<0.05). Poly‐ubiquinated protein levels were not different between any of the age groups.

CONCLUSION

It appears that plantaris muscle atrophy occurs with aging. However, this does not appear to be reflected via decrements in MPS or increases in protein polyubiquination. Mechanisms related to fast‐twitch skeletal muscle atrophy remain to be determined and should be further investigated.

Support or Funding Information

This study was supported in part by a contract from The University of Tampa (to M.D.R. from J.M.W.) as well as laboratory start‐up funds from M.D.R.
Enhancement of neural regeneration and functional recovery via DAPK1 suppression in stroke animal model
Authors: Won Jinyoung , Hong Yonggeun
Published: November 2023

Stroke is the most frequent worldwide cause of adult death. Recently, studies reported an intimate relation between DAPK1 as key player in multiple cell death and ischemic brain injury. We investigated the effect of DAPK1 on neural regeneration and functional recovery following ischemic injury using by Oxazolone, a synthesized DAPK inhibitor. In ischemic stroke model, subjected animals exhibited neurologic damage and altered behavioral outcome. However, suppression of DAPK1 enhanced neural regeneration and functional recovery. To analysis of neuronal morphology including soma, synapse number, and synaptic recruitment, Golgi‐cox staining was performed. Suppression of DAPK1 induced loss of dendrites length and spine density of pyramidal neurons, and supported progressive improvement of neurological deficits compared to control (P < 0.05). In addition, we investigate whether brain injury was accompanied by changes in levels of BDNF. Expression of immature precursor pro‐BDNF, involved in neuronal apoptosis, was decreased in response to ischemic injury under DAPK inhibited condition (P < 0.05). These results indicate that activation of DAPK1 following the ischemic insult exclusively correlate with brain injury and involve disturbed neurogenesis. Therefore, regulation of downstream molecules in DAPK1 pathway induces neural protection against stroke.

Support or Funding Information

Funding: NRF‐2012R1A1A2005089, NRF‐2013R1A2A2A01067169
Effects of Intraperitoneal Administration of Leptin on Voluntary Feed Intake, Appetite Signaling Pathways and Metabolism in Atlantic salmon, Salmo salar
Authors: Rønnestad Ivar , Søyland Marcus AL , Hansen Tom , Jordal Ann‐Elise O , Nilsen Tom Ole , Gomes Ana S , Björnsson Björn Thrandur , Jönsson Elisabeth Bergman , Hevrøy Enst Morten
Published: November 2023

Atlantic salmon is commercially a key species in the global aquaculture industry, and serves as a model species for teleost breeding, particularly regarding physiology and nutrition. Salmon is, however, sensitive to handling and solitary tank environment, and will respond to such stressors with reduced feed intake for extended periods. Therefore, we have validated an experimental protocol that incorporates a tank‐based system for analysis of individual feed intake in Atlantic salmon (N=16). It includes the use of sedation AQUI‐S prior to anaestesia (MS 222), allowing handling and experimental treatments like intraperitoneal (IP) injections. The protocol ensures low stress response and in the experiments described below, the salmon resumed voluntary feeding within 1–2 h of IP injections.

The effects of leptin (Lep)on voluntary feed intake and appetite in post smolt Atlantic salmon was studied. Three experiments were conducted alltogether where individual salmon were placed in tanks and feed intake monitored until it stabilized at a high level, after which each individual was IP injected with recombinant salmon leptinA1 (rsLepA1) emulsified in vegetable oil. The fish were restocked into the tanks and feed intake (hand‐feeding 4 times daily) was monitored for up to 4 more days. For all experiments, fish were sampled 4 h after the last meal and samples collected from brain, liver and stomach for qPCR analysis of appetite‐ and growth‐related genes (lepa1, 2; lepr1, 2; pomca1, a2, 2s, b; agrp1, 2; cart; npy; pyy; cckl). Plasma was collected and analysed for levels of the energy‐related metabolites triglycerides, glucose, free fatty acids, lactate and D‐3‐hydroxybutyrate.

Lep caused a significant reduction in feed intake for up to 3 days as well as a reduction in specific growth rate, compared to pre‐injection and sham controls. Brain expression of pomca1 and pomca2 was significantly (p<0.05) upregulated in the Lep‐treated fish. This supports earlier studies where pomc expression is linked to reduced appetite and feed intake. Brain pyy expression was significantly (p<0.05) down‐regulated in fish with reduced feed intake, sugging that PYY may be involved in the central Lep signaling pathway. Hepatic lepa1 and lepa2 mRNA expression was up‐regulated in Lep‐treated fish, but no significant differences in energy related metabolites in the plasma were found. In summary, a validated IP administration protocol for rsLepA1 has been established, demonstrating regulatory effects of homologous Lep on voluntary feed intake in Atlantic salmon.

Support or Funding Information

Research Council of Norway (grant no. 199683) and European Commission (FP7/2007‐2013; grant no 222719 – LIFECYCLE)
LIFE AND CONSCIOUSNESS AS QUALITIES
Authors: Ghosh Santibrata
Published: November 2023

It may be presumed in this work that at the moment of death, everything in the body ( viz. molecules and their motions ) remains identical to what it was at the last moment of life, the difference being that life has something more than the basic underlying physical and chemical elements. It is a totality, which is more like a flow, (i.e. a time derivative, or differential, as in Zeno's paradox of “an arrow in flight”). This is what may be called, in a dialectical sense, a new quality that is ‘life’. A similar picture was presented about consciousness, which is a function of the central nervous system ( not the autonomic ). In both, we assumed the movement of an arrow as being at point at an instant and not there the next instant.

The application of science in the hierarchy of life and consciousness is currently one of the most active areas of research in viewing the topic from the bottom‐up; i.e. chemistry >biology>neurology > consciousness. This is as opposed to a top‐down, philosophical and/or phenomenological approach to consciousness.

Here we show that both ‘life’ and ‘consciousness’ belong to a category called ‘phenomenon’, both lost instantly and may also represent a new ‘quality’ in the ‘dialectical’ sense. Based on this approach, transformations in nature can be classified into the types:

A union of opposites that can generate a new quality, as in the case of an arrow in flight.

Quantity to quality; random to organized, viz. molecules to life without consciousness; is reducible.

Mechanical to dialectic with transcendental properties; life with consciousness; consciousness from neural inputs; qualitative; is irreducible.

Support or Funding Information

none
Effects of chronic type 1 diabetes mellitus on structural changes, fibrosis, apoptosis and hypertrophy in the rat kidney
Authors: Singh Jaipaul , Bidasee Keshore R , Adehgate Ernest , Howarth Chris , Iqbal Tahreem
Published: November 2023

Diabetes mellitus (DM) is a major metabolic disorder and if left untreated can lead to diabetic nephropathy (DN). This study investigated the effects of 2 and 4 months of type 1 diabetes on the morphology, fibrosis and apoptosis in kidneys using biochemical and molecular techniques. The results show that DM induced significant (p<0.05; Student's t–test) changes in body weight. DM also increased blood glucose, kidney weight and kidney to body weight ratios, an indication of renal hypertrophy compared to controls. DM also induced structural changes in the kidneys including dilation of tubules, thickened of basement membrane and disorganization of the glomeruli. Interstitial and perivascular fibrosis were also prominent as was the development of apoptosis compared to controls. These changes were accompanied by significant (p<0.05) increases in the gene expressions of collagen 1‐alpha, collagen 3 alpha, fibronectin, elastin, MMP9, TIMP4, CTGF, connexin 43, integrin alpha 5, vimentin TGF‐beta1, BNP and ANP compared to controls. Changes were more pronounced and significantly higher (p<0.05) in older DM rats. The results reveal that DM can be detrimental to the kidney structure and function leading to DN.
miR‐150 regulates adipose tissue B cell function by targeting the BCR signaling pathway
Authors: Wang Haiqing , Tseng Alexander , Chang Richard Cheng‐An , Lin Yu‐lieh , Ying Wei , Zhou Beiyan
Published: November 2023

Metainflammation and insulin resistance are two hallmarks of obesity which contribute to the pathogenesis of obesity‐associated diseases, including type 2 diabetes and cardiovascular diseases. Expansion of visceral adipose tissue (VAT) is central to the development of obesity associated metabolic syndromes, characterized by adipocyte malfunction and altered tissue specific immune cell profiles. Adipose tissue immune cells vary in number and their responses to obese stress. The adipose tissue resident B cells population dramatically expands to approximately 20% of stromal cells within obese visceral adipose tissues; however, their functions in the adipose tissue niche are poorly elucidated.

Here we report that miR‐150 modulates adipose tissue function by controlling activation of B cells and their interactions with other immune cells. miR‐150KO mice displayed exacerbated obesity‐associated tissue inflammation and systemic insulin resistance, which is recapitulated by adoptive transfer of B cells, but not purified immunoglobulin, into obese Bnull mice. Using purified cell populations and a co‐culture platform, we found that miR‐150KO B cells directly enhanced proinflammatory activation of adipose tissue T cells and macrophages through cell‐cell interactions, but not through cell‐autologous mechanisms. Furthermore, miR‐150KO B cells displayed significantly enhanced antigen presentation upon stimulation, ultimately leading to elevated inflammation and insulin resistance, compared to wild type B cells. Target gene prediction analysis revealed that several targets of miR‐150, Elk1, Etf1 or Myb, are components of the B cell receptor (BCR) signaling pathway. We validated these genes as bona fide miR‐150 targets and discovered that knockdown of their expression attenuated B cell action and MHCII dependent antigen presentation. Thus, our results demonstrate a critical role for a miR‐150/BCR signaling pathway regulatory axis within adipose tissue B cells which ultimately regulate both adipose tissue metabolic and immunologic homeostasis.

Support or Funding Information

American Diabetes Association (1‐13‐JF‐59 to B. Zhou)National Institute of Health/National Institute of Diabetes and Digestive and Kidney Diseases (NIH/NIDDK 1R01DK098662 to B. Zhou)
Palmitoyl‐carnitine production by blood cells is associated with plasma palmitate oxidation in women
Authors: Chondronikola Maria , Asghar Rabia , Zhang Xiaojun , Dillon Edgar L , Durham William J , Wu Zhanpin , Porter Craig , Camacho‐Hughes Maria , Zhao Yingxin , Brasier Allan R , Volpi Elena , Sheffield‐Moore Melinda , Abate Nicola , Sidossis Labros , Tuvdendorj Demidmaa
Published: November 2023

Circulating acyl‐carnitines (acyl‐CNTs) are associated with insulin resistance (IR) and type 2 diabetes (T2D) in both rodents and humans. However, the mechanisms whereby circulating acyl‐CNTs are increased in these conditions and its implications for whole‐body metabolism have not been determined. Here we aimed to determine if blood cells contribute in production of circulating acyl‐CNTs and whether such production is correlated with whole‐body fatty acid fluxes.

Methods and Results

Eight non‐diabetic healthy women (Age: 47±19 y; BMI: 26±1 kg•m−2) underwent stable isotope tracer infusion and hyperinsulinemic‐euglycemic clamp study to determine in vivo whole‐body fatty acid kinetics and insulin sensitivity. At baseline (0 min) and after 3 h of hyperinsulinemic‐euglycemic clamp, blood samples were collected for measurement of in vitro synthesis rates of palmitoyl‐carnitine (palmitoyl‐CNT). The fractional synthesis rate of palmitoyl‐CNT was significantly higher during hyperinsulinemia (0.788±0.084 vs. 0.318±0.012 %•hr−1, p=0.001); however, the absolute synthesis rate did not differ between the periods (p=0.809) due to decreased (~30%) palmitoyl‐CNT concentration (p=0.189) during hyperinsulinemia. Regression analyses demonstrated that the absolute synthesis of palmitoyl‐CNT significantly correlated with the concentration of acyl‐CNTs in basal (r=0.992, p<0.001) and insulin (r=0.919, p=0.001) periods; and the basal absolute synthesis significantly correlated with plasma palmitate oxidation rate (r=0.764, p=0.027).

Conclusion

In women, blood cells contribute to plasma acyl‐CNT levels and basal whole‐body plasma palmitate oxidation is associated with acyl‐CNT production. Future studies investigating the role of blood cells in acyl‐CNT and lipid metabolism under different physiological (i.e., hyperlipidemia) and pathological (i.e., IR and T2D) conditions are warranted.

Support or Funding Information

Claude D. Pepper OAIC grant (# P30‐AG024832), the Institute for Translational Sciences at the UTMB, supported in part by a Clinical and Translational Science Award (#UL1 TR001439) from the NCATS/NIH, and the Shriners Grant #84090, Metabolism Unit, Shriners Hospitals for Children.
Human Physiological Performance During a 1‐Kilometer Swim in Cold Water
Authors: Cummings Katelyn Marie , Olson Mariah F , Treu Spencer P , Bute Derek M , Malone Christopher J , Barkai Ram , Wilson Ted
Published: November 2023

Human physiological responses to cold‐water swimming (≤5°C) are poorly characterized. A database (www.internationaliceswimming.com) describes 1‐kilometer ice swimming performance times and swim conditions. The present study analyzed data from 73 males and 30 females who completed 1‐kilometer ice swims in water ≤5°C. Swim completion time was 18.77 ± 3.63 minutes for males and significantly slower for females at 20.93 ± 4.89 minutes (P = 0.0042). Swim time decreases as age increases by 0.19 minutes per year for males; whereas the rate of decrease is 0.29 minutes per year for females, suggesting that there is a relationship between age and gender (P = 0.0209). Air temperature also statistically impacts swim time when adjusting for age and gender (P = 0.0079). In summary, ice swimming is increasing in popularity and is being considered for future Winter Olympic activities. Characterization of cold‐water exposure physiology remains an important consideration for emergency, military, and medical personnel. The present study provides a useful benchmark for understanding and predicting physiological performance during exposure to cold environmental conditions.
Mitogen‐Activated Protein Kinases Differentially Regulate Renal Cell Paracellular Permeability
Authors: Watari Jessica , Axis Josephine , Jaggi Shirin , Voronina Angelina , Amsler Kurt
Published: November 2023

Movement of compounds between renal epithelial cells, paracellular permeability, is mediated primarily by the permeability of the circumferential tight junction structure (TJ) located at the apicolateral border of adjacent epithelial cells. Hydrogen peroxide (H2O2), which contributes to renal ischemia‐reperfusion injury, increased the paracellular permeability of two renal epithelial cell lines, MDCK (distal tubule) and LLC‐PK1 (proximal tubule), in a concentration‐dependent manner. H2O2 treatment also produced a concentration‐dependent increase in the phosphorylation of the Mitogen‐Activated Protein Kinases ERK 1/2, JNK 1/2, and p38MAPK. Pretreatment of renal cell populations with inhibitors of ERK 1/2 activation (U0126, PD98059) and p38MAPK activity (SB202190), diminished or completely blocked the ability of H2O2 to increase paracellular permeability. In contrast, treatment with two JNK 1/2 inhibitors, SP600125 and AS601245, did not block the ability of H2O2 to increase renal paracellular permeability. In fact, treatment with JNK 1/2 inhibitors increased paracellular permeability in a concentration‐dependent manner when added alone and augmented the increase in paracellular permeability when added prior to H2O2 treatment. Previous studies demonstrated that treatment of renal epithelial cells with H2O2 slowed the movement of GFP‐occludin into the TJ region, as measured by Fluorescence Recovery After Photobleaching. Treatment of cell populations with U0126 alone slowed the movement of GFP‐occludin into the TJ region. In U0126‐treated cell populations, however, H2O2 treatment was unable to slow further the rate of GFP‐occludin movement into the TJ region. These results indicate: 1) H2O2 treatment increases renal epithelial paracellular permeability via activation of ERK 1/2 and p38MAPK; 2) Steady state JNK 1/2 activity may promote maintenance of the paracellular permeability barrier; and 3) The ability of H2O2 to modulate the mobility of occludin into the TJ structure may be associated with its ability to increase renal cell paracellular permeability.

Support or Funding Information

Research supported by NIH grant R15 DK091749
Peripheral Apelin Inhibits Gastrointestinal Motor Functions via CCK‐dependent Pathway
Authors: SİNEN Osman , BÜLBÜL Mehmet , İzgüt‐Uysal V. Nimet
Published: November 2023

Apelin is the endogenous ligand of the G protein‐coupled receptor APJ. The apelinergic system and APJ receptors are widely expressed in gastrointestinal (GI) tissues including stomach, small intestine and colon, suggesting that peripheral apelin may have a regulatory role in GI motor functions. It has been shown that peripheral exogenous administration of apelin increases the release of cholecystokinin (CCK), a well‐defined GI hormone that disturbs gastric emptying (GE), colon transit (CT) and small intestinal fasting motor pattern through CCK‐1 receptors expressed on vagal afferent nerves. The present study was designed to investigate (1) the effects of peripherally administered apelin on GI motor functions and (2) the contribution of CCK in apelin‐induced alterations. For measuring solid GE, rats were given a 1.6 g of preweighed pellet following an overnight fasting. Rats were euthanized 90 min after the completion of feeding and the gastric contents were recovered, dried, and weighed. To measure CT, a silicone catheter was inserted into the proximal colon and fixed with silk sutures; whereas, a strain gage transducer was implanted on the serosal surface of duodenum for recording of intestinal fasting motor pattern. The catheter and the wires of transducer were tunneled subcutaneously and secured to the neck skin. The measurements and recordings were performed after a 7‐day recovery period. For CT measurement, 1.5 ml of phenol red solution was injected through the catheter and rats were euthanized 90 min after injection. The entire colon was removed and divided into 6 equal segments. CT was calculated as the geometric center of distribution of phenol red. In control rats, GE was found 60.1 ± 3.5%; whereas, apelin‐13 administration (100 μg, ip) significantly (p<0.05) delayed the gastric emptying 31.3 ± 2.8%. The apelin‐induced delayed GE was significantly (53.3 ± 5.8%, p<0.05) accelerated by pretreatment of CCK‐1 receptor antagonist lorglumide (2.5 mg, ip) and restored to the control level. Compared with control rats (4.1 ± 0.1), apelin‐13 significantly inhibited CT (3.2 ± 0.1%, p<0.01). However, pretreatment of lorglumide significantly attenuated (4.1 ± 0.1, p<0.05) the inhibitory effect of peripheral apelin administration. The fasting‐induced duodenal phase‐III‐like contractions were remarkably disturbed following apelin‐13 administration. However, pretreatment of lorglumide abolished the apelin‐induced alterations in intestinal fasting motor pattern. These findings suggest that peripheral apelin inhibits GI motor functions and the inhibitory effects of apelin appear to be CCK‐dependent. Peripheral apelin may have a regulatory role in postprandial changes in alimentary motor functions.

Support or Funding Information

This work is funded by The Scientific and Technological Research Council of Turkey (TUBITAK). (Research Programme 1001; project number 214S349).