bluuebunny/crossref_metadata_2025_split huggingface dataset:
Isoprenoid Depletion Increases Cytokine mRNA Levels in Primary Human Monocytes
Authors: Frey Tiffany Albright 
Published: June 2021

Mevalonate Kinase Deficiency (MKD) is an autoinflammatory disease caused by mutations in the mevalonate kinase (MVK) gene, which produces an enzyme responsible for the production of isoprenoids. Previous research in various model systems has demonstrated that depletion of isoprenoids leads to inflammasome activation and increased levels of secreted IL‐1B on a post‐translational level. However, patient data indicates that MKD is a multi‐cytokine disease with increased plasma levels of cytokines such as TNF‐alpha, IL‐6, and IFN‐gamma. In order to study the mechansims responsible for these changes, the mevalonate pathway was inhibited with lovastatin in peripheral blood mononulcear cells (PBMCs) and monocytes isolated from the blood of healthy donors followed by stimulation with lipopolysaccharide (LPS) to induce an inflammatory response. Lovastatin treatment resulted in significantly increased levels of TNF‐alpha, IL‐6, IL‐12p40, and IFN‐gamma mRNA in both PBMCs and monocytes following LPS stimulation as compared to control cells. Lovastatin treatment did not alter IL‐1B, CD14, or IL‐18 mRNA levels either before or after LPS stimulation. Addition of mevalonate to the cells treated with lovastatin prevented the increased cytokine mRNA levels indicating that these changes are due to inhibition of the mevalonate pathway. These results indicate that isoprenoid depletion alters the expression of multiple cytokines on the mRNA level in primary human monocytes, which can be used as a model system ex vivo to study these changes.Support or Funding InformationDickinson College Faculty Start‐up and Research & Development Committee FundingThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Chronic alcohol misuse induces alveolar macrophage oxidative stress and mitochondrial dysfunction via upregulation of microRNA‐130a
Authors: Yeligar Samantha M. 
Published: June 2021

ObjectiveCompared to non‐alcoholics, individuals who misuse alcohol are 2–4 times as likely to develop respiratory infections and acute respiratory distress syndrome. Alcohol misuse enhances alveolar macrophage (AM) oxidative stress and causes phagocytic dysfunction via upregulation of microRNA (miR)‐130a. Although studies indicate that alcohol‐induced mitochondrial (MT) redox imbalance in AM may contribute to AM phagocytic dysfunction, the mechanisms by which this occurs is unknown. We hypothesized that alcohol upregulates miR‐130a to destabilize its target peroxisome proliferator‐activated receptor (PPAR)γ, which dysregulates AM MT function and depletes ATP levels necessary for phagocytosis.MethodsAMs were obtained from the bronchoalveolar lavage fluid of C57BL/6J mice fed ± ethanol (20% w/v) in the drinking water for 12 wks. In parallel, MH‐S cells, a mouse AM cell line, were transfected ± 50 nM miR‐130a inhibitor and treated ± 0.08% ethanol for 3 d. Levels of PPARγ‐related miR‐130a, which binds to the PPARγ 3′untranslated region to decrease PPARγ, and PPARγ mRNA were assessed by qRT‐PCR. PPARγ protein expression was assessed by confocal cytoimmunostaining. MT‐derived oxidative stress was measured using MitoSOX and MitoTracker CMXRos fluorescent molecular probes. MT bioenergetics profile was measured with MT stress testing using a Agilent Seahorse XF96 Extracellular Flux Analyzer. AM function was evaluated by phagocytosis assay (S. aureus internalization).ResultsIn vivo and in vitro studies demonstrated that ethanol: 1) increased miR‐130a levels and decreased PPARγ expression; 2) increased MT‐derived oxidative stress; 3) decreased MT oxygen consumption rate, bioenergetics, and ATP; and 4) impaired phagocytic capacity. miR‐130a inhibitor reversed these ethanol‐induced AM derangements.ConclusionsChronic alcohol misuse enhances AM oxidative stress and MT dysfunction by upregulating PPARγ‐related miR‐130a. Our studies suggest that therapeutically targeting MT as an intervention will ameliorate alcohol‐induced AM phagocytic dysfunction and may decrease susceptibility of lung infections in patients with a history of alcohol misuse.Support or Funding InformationResearch Funding Source: R00AA021803 (SMY).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Patterns of Suppressed Mitochondrial Respiration in Isolated Muscle Fibers from Type 2 Diabetics
Authors: Turner Kristen D. , Harris Matthew P. , Kronemberger Ana , Ueda Kenichi , Feider Andrew J. , Kenny Helena , Anderson Ethan J. , Abel E. Dale , Casey Darren P. , Lira Vitor A. 
Published: June 2021

The quality of skeletal muscle is markedly influenced by mitochondrial function. Aberrant mitochondrial function has been associated with reduced contractile and metabolic dysfunction in skeletal muscle, potentially leading to insulin resistance and type 2 diabetes. However, the direct relationship between mitochondrial dysfunction and type 2 diabetes remains to be fully understood. In the present study, vastus lateralis muscle biopsies were obtained from individuals with type 2 diabetes (N = 7) and non‐diabetic, age, gender and weight‐matched controls (N = 7). Maximal mitochondrial oxygen consumption was assessed in isolated muscle fibers and normalized to wet tissue weight. Preliminary findings illustrate that reductions in carbohydratedependent ADP‐stimulated respiration (~30%) were sustained in muscle fibers from type 2 diabetics, even with the addition of the Krebs Cycle anaplerotic substrates, glutamate (~30%) and succinate (~32%). Interestingly, trends of similar magnitudes persisted for lipid substrate metabolism, as fatty acid‐dependent ADP‐stimulated respiration also tended to be lower (~35%) compared to controls. Furthermore, rotenone‐treated respiration in diabetic skeletal muscle tended to be reduced (~37%). To conclude, these initial findings provide support for a systematic impairment in the oxidative metabolism of both carbohydrates and lipids in skeletal muscle of type 2 diabetic individuals. In addition, findings from rotenone‐treated conditions indicate that investigating functional deficits of Complex I of the Electron Transport Chain (ETC) may provide a more insightful understanding of the relationship between mitochondrial respiratory control and type 2 diabetes.Support or Funding Information*This study is supported by funds from the American Diabetic Association, ADA‐1‐16‐ICIS‐015, (D.P.C.) and start‐up funds from the Department of Health and Human Physiology at the University of Iowa (V.A.L.).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Comparison of Different Mathematical Approaches to Assess Calcium Retention Capacity in Cardiac Isolated Mitochondria following Ischemia Reperfusion Injury
Authors: Lahil Harjeev S. , Salzman Michele M. , Balzer Claudius , Cleveland William J. , Riess Matthias L. 
Published: June 2021

A hallmark of myocardial ischemia reperfusion (IR) injury is mitochondrial (m) dysfunction resulting in, e.g., altered electron transport, increased reactive oxygen species formation, decreased adenosine triphosphate synthesis and reduced calcium (Ca2+) retention capacity (CRC). Different cardioprotective strategies aim to attenuate this dysfunction. CRC is typically assessed by adding boluses of extra‐mCa2+ which is taken up by functioning mitochondria to a higher degree, leading to a later opening of the mitochondrial permeability transition pore (mPTP) and release of accumulated intra‐mCa2+ than by damaged mitochondria. We hypothesized that mPTP opening could be predicted by the degree of decrease in the rate of mCa2+ uptake after each exogenous Ca2+ bolus before the mPTP opens. Following IR with and without different cardioprotective stratetgies, mitochondria were isolated from Brown Norway rat (n=38) and Arctic Ground Squirrel (n=19) isolated hearts, and CRC was assessed by measuring changes of extra‐mCa2+ by calcium‐green fluorescence when repeated boluses of exogenous Ca2+ were given in the presence of pyruvate/malate or succinate as complex I or II substrates, respectively. Multiple regression analyses (alpha 0.05) were used to compare the actual number of Ca2+ boluses leading to mPTP opening to the number predicted from absolute mCa2+ uptake and %mCa2+ uptake, respectively. There were no significant differences in regressions between species. Despite acceptable regression coefficients (R2: 0.81/0.81 vs 0.76/0.78), both methods (absolute/%) had a regression slope significantly higher than 1.0 (slope±SEM: 1.28±0.08/1.35±0.09 vs 1.72±0.13/2.00±0.14) for complex I vs II substrates, respectively. This indicates an increasing overestimation of CRC by either method compared to the actual number of boluses before mPTP opening, more so in less dysfunctional compared to more dysfunctional mitochondria. This overestimation was stronger in the presence of complex II vs I substrates. Thus, predictions of eventual mPTP opening from mCa2+ uptake after individual exogenous Ca2+ boluses are more reliable in dysfunctional vs functional mitochondria and when complex I substrate is used compared to complex II substrate.Support or Funding InformationMerit Review Award (IK2 BX001278 and I01 BX003482) from the United States Department of Veterans Affairs Biomedical Laboratory R&D Service; National Institutes of Health (5R01 HL123227) and institutional funds.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
ANP antibody attenuates RIHP amplification during acute Saline Volume Expansion in rats
Authors: Briones María Eugenia Sánchez , Lopez‐Rodríguez Juan Francisco , Calvo‐Turrubiartes Miriam Z. , Rodríguez‐Martínez Manuel 
Published: June 2021

The role of Atrial Natriuretic Peptide (ANP) on renal interstitial hydrostatic pressure (RIHP), urinary excretion of sodium (UNaV) and water (V) amplification responses during saline volume expansion (SVE, 5% bw, ~30 min, 0.9%), and immediately after (maintained SVE, 10 min) was studied in male adult wistar rats 24 h after antiseptic intra‐arterial infusion of polyglonal IgG anti‐Atrial Natriuretic Factor (ANF) antibody (AbANP, 18 μg, wich theoretically blocked all ANP secretion, n=8) versus control (CTR, gluc5%, n=10). The day of the acute experiment, variables were continously measured in anesthetized animals with hormonal clamp (without ANP), kidney denervation (chemical and mechanical) and renal perfusion pressure control at 100 mmHg during hydropenia (H), SVE, and maintained SVE(mSVE).The results are shown in table 1. For Ht and plasmatic protein, there were no differences between groups during H period and the hemodilution response was similar between groups. For RIHP, an attenuation in the amplification response was obtained, from 3 times in CTR vs H to 1 time in abANP group (MANOVA test, p<0.05 between groups, SVE and mSVE., t‐student test vs H each group p<0.05). For V and UNaV responses, there were no differences between groups or periods, achieving the amplification response in the same magnitude for both groups. Finally, an increase in UGMPcV and UNOxV during SVE and mSVE were similar in both groups.Conclusionsin rats, the acute increase in plasmatic ANP is necessary for the complete RIHP amplification during acute SVE, probably involving an independent nitric‐oxide mechanism. Also, a dissociation between RIHP and natriuretic response was observed.Support or Funding InformationSupported by IPL's financial resourcesThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Renal protective effects of the flavonoid (−)‐Epicatechin in a chronic kidney disease (nephrectomy 5/6) model
Authors: Montes Osvaldo , Ramirez‐Sanchez Israel , Villarreal Francisco , Meaney Eduardo , Najera Nayelli , Arellano Monica , Ceballos Guillermo 
Published: June 2021

BackgroundChronic kidney disease (CKD) is a worldwide public health problem affecting 10% of the adult population. One of the most important factors in the progression of CKD from early stages is oxidative stress. Currently there is limited efficacy to treat this problem so the objective of this study was to test Epicatechin (Epi), effects in a mice model of CKDMethodsA nephrectomy 5/6 (Nf) in C‐57BL6 mice model was implemented and Epi treatment (14 days doses doses of 0.01, 0.1 and 1 mg/kg) was provided by gavage. Changes in Systolic blood pressure (SBP), serum urea and creatinine, renal cortex morphology, serum tetrahydrobiopterin (BH4), in 3‐nitrotyrosine as oxidative damage marker and of Angiotensin II AT1 receptor (AGTR1) and NOX‐4 protein expression were evaluatedResultsSBP increased in the Nf group and decrease significantly in Nf+ Epi groups. Serum urea and creatinine levels increased in the Nf group Epi treatment induced a significant decrease. Epi treatment induced a dose dependent increase in serum BH4 in Nf animals. Normal renal cortex morphology was conserved at 1 m/kg Epi dose. The expression of AGTR1 and NOX‐4 was increased in the Nf group, Epi treatment in a dose dependent manner decrease the AGTR1 and NOX‐4 expression.ConclusionThe present findings suggest that Epi could prevent the progression of CKD and may be used as complement for the modulation of blood pressure and f renal dysfunction.Support or Funding InformationConacyt 253769This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Renal Expression of Adhesion GPCR Gpr116 (ADGRF5) Plays a Role in Urinary Concentration in Mice
Authors: Zaidman Nathan , Pluznick Jennifer 
Published: June 2021

G‐protein coupled receptors (GPCR) are a large and diverse family of integral membrane proteins that recognize a tremendous assortment of extracellular molecules including neurotransmitters, hormones, light and odors. They are a common target of pharmaceutical drug development, and uncovering the function of novel GPCRs in the kidney represents a wealth of untapped therapeutic potential. We previously performed an mRNA screen for novel GPCRs in the kidney to identify promising yet overlooked renal GPCRs. This screen revealed that Gpr116, an adhesion‐class GPCR, is highly expressed in the kidney. To understand the role of Gpr116 in renal physiology, we have taken a multidisciplinary approach. Using transfected HEK293 cells expressing cloned Gpr116, we have validated a monoclonal antibody for Gpr116. In murine kidney, this antibody indicates localization of Gpr116 to intercalated cells in the collecting duct of mouse kidneys. This staining is absent in kidneys from targeted knockout (KO) of Gpr116 in renal tubules (Gpr116flox/flox, ksp‐Cre). Additionally, kidney‐specific KO animals have significantly reduced urine osmolality (1552±71 mOsm/kg, N=11) compared to wild‐type (WT) (2097±76, N=10, p<0.0001) and heterozygous (2423±86, N=6, p<0.0001) littermates. Lastly, preliminary data of urine osmolality following 12‐hour water restriction (dark‐cycle) suggests KO animals are capable of concentrating urine, though only to about 75% that of WT littermates (3492±150 vs. 4361±525; N=3, N=4, respectively). Overall, these results suggest a role for Gpr116 in enhancing the ability to concentrate urine. This study establishes a physiologic role of the previously understudied Gpr116 in the murine kidney and demonstrates the scientific potential of future investigations into novel GPCRs.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Kir4.1 activity is essential for dietary Na+ intake induced modulation of Na‐Cl cotransporter (NCC)
Authors: Su Xiao‐Tong , Wu Peng , Gao Zhongxiuzi , Wang Wen‐Hui , Lin Dao‐Hong 
Published: June 2021

Dietary Na+ intake has been shown to modulate the thiazide‐sensitive NCC in the distal convoluted tubule (DCT), such that a high Na+ intake (HNa) inhibits whereas a low Na+ diet (LNa) stimulates NCC activity. However, the mechanism by which dietary Na+ intake on NCC is not clear. The aim of the present study is to test the hypothesis Kir4.1 is essential for mediating the effect of dietary Na+ intake on NCC. The patch‐clamp experiments demonstrated that the basolateral 40 pS K+ channel (a Kir4.1/Kir5.1 heterotetramer) activity in the DCT was upregulated in the mice on a LNa diet for 7 days but it was down‐regulated in the mice on HNa for 7 days. The whole‐cell recording further demonstrated that HNa decreased basolateral K+ conductance of the DCT and depolarized the membrane. Conversely, LNa intake augmented the basolateral K+ currents and hyperpolarized the DCT membrane. Western blot and renal clearance studies demonstrated that LNa increased the expression of total NCC (tNCC) and phosphorylated NCC (pNCC) and enhanced hydrochlorothiazide (HCTZ)‐induced natriuresis while HNa intake had opposite effects on the expression of tNCC and pNCC and reduced HCTZ‐induced natriuresis. Thus, an upregulation of Kir4.1 is correlated with a high NCC activity in the mice on LNa while a down‐regulation of Kir4.1 is related with low NCC activity in the mice on HNa. To explore the role of Kir4.1 in mediating the effect of dietary Na+ intake on NCC, we repeated the above experiments in kidney‐specific conditional Kir4.1 knockout mice (KS‐Kir4.1 KO). The deletion of Kir4.1 completely abolished the basolateral K+ conductance and depolarized the DCT membrane. Moreover, the effect of dietary Na+ intake on the basolateral K+ conductance and membrane potential was also absent in KS‐Kir4.1 KO mice. Also, the deletion of Kir4.1 decreased the expression of both tNCC and pNCC and abolished HCTZ‐induced natriuresis. Western blot and renal clearance experiments showed that the effect of dietary Na+ intake on NCC expression and activity was absent in KS‐Kir4.1 KO mice. We conclude that Kir4.1 plays an essential role in mediating the effect of dietary Na+ intake on NCC activity such that LNa‐induced stimulation of Kir4.1 in the DCT is essential for the upregulation of NCC while HNa‐induced inhibition of Kir4.1 is required for the down‐regulation of NCC.Support or Funding InformationNIH DK54983This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Vasopressin V1a Receptor of Renal Collecting Duct Intercalated Cells Mediates Urinary Acidification
Authors: Giesecke Torsten , Himmerkus Nina , Leipziger Jens , Isermann Julian , Ayasse Niklas , Koshimizu Takaaki , Kawahara Katsumasa , Gimber Niclas , Schmoranzer Jan , Bleich Markus , Fähling Michael , Smorodchenko Alina , Bachmann Sebastian , Mutig Kerim 
Published: June 2021

Vasopressin (AVP) drives urinary concentration via V2 receptors in the distal nephron and collecting duct of the kidney. Parallel activation of V1a receptors (V1aR) has been implicated in renal acid‐base handling, but cell type‐related information on this receptor is still scarce. We hypothesized that activation of V1aR in intercalated cells (IC) contributes to acid‐base homeostasis by the kidney.We generated an anti‐V1aR antibody and verified its specificity using V1aR‐knockout tissues. For localization studies in mice, rat and human tissue, we used immunofluorescence and confocal microscopy or 3D‐structured illumination microscopy (3D‐SIM). For functional studies in vivo, V1aR‐specific agonist, AO‐4‐67, was administered in AVP‐deficient Brattleboro rats and C57/BI6J mice (0.2, 2, or 10μg/kg b.w.; 1 to 4h). Urine was collected in metabolic cages or via ureteral catheter and plasma samples were taken. Isolated, microperfused mouse collecting ducts were analyzed for V1aR‐dependent luminal pH changes and cultured inner medullary collecting duct cells for proton‐secreting vH+‐ATPase regulation.V1aR immunoreactivity of type‐A IC was demonstrated in basolateral membrane and cytoplasmic granules, whereas type B IC revealed perinuclear and subapical granular signal alone. Administration of AO‐4‐67 to Brattleboro rats for 4h resulted in increased urinary proton secretion (+34% after 1h, p<0.05) and decreased bicarbonate excretion (up to −90% after 3h, p<0.001). Similarly, catheterized mice showed rapid urinary acidification in response to AO‐4–67 (pH from 7.18 to 6.78 after 20 min, p<0.05). Basolateral application of V1aR agonist or AVP in isolated perfused collecting ducts resulted in decreased luminal pH values (−5% after 3 min, p<0.05). Cultured collecting duct cells showed significant increases in luminal vH+‐ATPase immunoreactivity upon V1aR stimulation (+90% after 4h, p<0.001).In summary, our results suggest that activation of V1aR in renal type‐A intercalated cells induces proton secretion into the urine and promote urinary acidification.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Interpretation of Whole‐Kidney RNA‐Seq Data
Authors: Clark Jevin , Neijman Kim , Chou Chung‐Lin , Deen Peter , Chen Lihe , Knepper Mark 
Published: June 2021

RNA sequencing (RNA‐Seq) is a sensitive means of global identification and quantification of mRNAs. In kidney, we have carried out RNA‐Seq analysis to identify transcriptomes of single microdissected renal tubules and single cells. The resulting datasets provide a community resource for gene expression in kidney structures in the form of a publicly accessible website at https://hpcwebapps.cit.nih.gov/ESBL/Database/Targets/TranscriptomicData.html. We now have added new whole‐kidney RNA‐Seq data for mice (male, age 8 weeks). This dataset reports 13,334 transcripts with mean TPM (“normalized transcripts per million”) greater than 1 for whole kidney (n=3). A large fraction of these are housekeeping genes or genes expressed in multiple cell types. Whole‐kidney RNA‐Seq analysis is of limited use in studying responses in these genes since the responses may be variable among cell types. However, signals were readily quantified for major cell‐type specific transcripts including podocytes (podocin, Nphs2), proximal convoluted tubule (sodium coupled glucose transporter 2, Slc5a2), proximal straight tubule (sodium coupled glucose transporter 1, Slc5a1), thin descending limb of Henle (follistatin, Fst), thin ascending limb (chloride channel ClC‐Ka, Clcnka), thick ascending limb (NKCC2, Slc12a1), distal convoluted tubule (NCC, Slc12a3), principal cell of collecting duct (ENaC gamma subunit, Scnn1g), type A intercalated cell (AE1, Slc4a1), type B intercalated cell (pendrin, Slc26a4), and inner medullary collecting duct cell (urea channel UT‐A1, Slc14a2 long isoform). Beyond this, transcripts were quantifiable for marker genes corresponding to rare cell types in the kidney including granular cells of the afferent arteriole (renin, Ren1), macrophages (macrophage colony‐stimulating factor 1 receptor, Csf1r), dendritic cells (dendritic cell‐specific ICAM‐3‐grabbing non‐integrin, Cd209a), and umbrella cells of the transitional epithelium (uroplakin 1a, Upk1a). In conclusion, numerous cell‐type specific mRNAs are represented in whole‐kidney RNA‐Seq analysis. These markers provide a means of measuring physiological regulatory responses (e.g. renin expression in granular cells) or measuring changes in cell numbers, which can occur with proliferative responses or infiltration of the kidney as part of inflammatory responses (e.g. macrophages and dendritic cells).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.