bluuebunny/crossref_metadata_2025_split huggingface dataset:
Activation of Mitochondrial Calpains Contributes to the Selective Degradation of Specific Mitochondrial Proteins
Authors: Chen Qun , Thompson Jeremy , Hu Ying , Hollander John M. , Lesnefsky Edward J. 
Published: June 2021

INTRODUCTIONCalpain 1 (CPN1) and 2 (CPN2) are ubiquitous calpains that exist in both cytosol and mitochondria. Calpains are calcium‐activated proteases. Pharmacologic inhibition of CPN1 and CPN2 decreases cardiac injury by improving mitochondrial function during ischemia‐reperfusion. Since both CPN1 and CPN2 include one large subunit and a small regulatory subunit (calpain 4, CPN4), genetic removal of CPN4 eliminates the activities of CPN1 and CPN2. CPN4 knockout mice were used in this study to test if activation of mitochondrial calpains leads to mitochondrial damage by proteolysis of mitochondrial proteins.HYPOTHESISWe hypothesize that exogenous calcium‐mediated activation of mitochondrial CPN1 and CPN2 impairs mitochondrial function through degradation of mitochondrial proteins.METHODSPurified cardiac mitochondria were isolated from wild type and CPN4 knockout mice. Exogenous calcium (25 uM) was used to activate mitochondrial CPN1 and CPN2. Mitochondria were subjected to proteomic analysis using tryptic digestion, liquid chromatography, and mass spectrometry with iTRAQ quantification.RESULTSWe focused on the proteins that were decreased by exogenous calcium exposure in mitochondria from wild type but not in CPN4 knockout mice as specific targets of mitochondrial CPN1 and CPN2. In wild type mice, calcium treatment significantly decreased the contents of multiple metabolic enzymes involving fatty acid oxidation and oxidative phosphorylation (electron transport chain components and ATP synthase) compared to non‐calcium treated mitochondria. Knockout of CPN4 preserved these enzyme contents in calcium‐treated mitochondria (Table). Decreases in ATPase factor 6 and very long chain acyl‐coA dehydrogenase were confirmed by Western blot.CONCLUSIONActivation of mitochondrial calpains leads to decreased mitochondrial function by impairing metabolic enzyme activity. Prevention of CPN1 and CPN2 activation provides an attractive approach to protect mitochondria during pathologic conditions including ischemia‐reperfusion and heart failure.Support or Funding InformationThis work was supported by the Office of Research and Development, Medical Research Service Merit Review Award (2IO1BX001355‐01A2) and Department of Veterans Affairs (EJL); National Institutes of Health NIH R21AG054975‐01 (QC).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Morphometric Evaluation of Gross Pulmonary Indices in the Red Sokoto Goats for Pulmonary Consolidation Assessment
Authors: Alaka Olugbenga Olayinka , Unuoah Precious Adetokunbo , Jarikre Theophillus Aghogho , Emipke Benjamin Obukowho 
Published: June 2021

Goats are widely adapted to different climates and are found in all production systems. However, very little is known on the pulmonary morphometry of indigenous goats in Africa. The study aims to determine the standard morphometry and dimensions of the Red Sokoto RS goat lungs, which may serve as basis for clinical and radiographic assessment of pulmonary consolidation. The goats were sourced from a Municipal abattoir considering age and sex. A total of 35 non‐pneumonic RS lungs were collected including 22 males and 8 females, 6 between <1 years, 15 between 1–2 years and 9 > 2 years. Weight (gram), length (cm) and diameter (cm) of each lung lobe, distance from trachea bronchus to point of trachea bifurcation, distance between left and right trachea bifurcation were taken using standard measurements. The right lung contributes the most (61.67%) while the left lung contributes the least weight (38.32%). Of the 61.67% contributed by the right lung, the caudal lobe contributes 38.9% while the cranial (14.71%), middle (5.83%) and accessory (1.53%) lobes makes up the remaining. It was also discovered that the caudal lobe of the right lung contributes more than the left lung. The percentage contribution of right lung lobes increased with increase in age except for the accessory lobe which decreased with increasing age. The percentage contribution of the left lung lobes decreased with increasing age except for the middle lobe which was increasing with age. The right lung is longer (6.70±0.57) than the left lung (13.40±0.82). The right cranial lobe is longer and wider than the left cranial lobe. The right middle lobe is insignificantly longer (1.20±0.57) than the left (1.10±0.22) but significantly less wide (6.7±0.57) than the left (7.72±1.14). The right caudal lobe is longer (8.76±0.25) than the left (8.50±0.50) but less wider (4.86±0.20) than the left (5.5±0.61). The males have higher weight and dimensions than females except that the values of the right lung, the caudal lobe of the right lung and the middle lobe of the left lung are higher in females than in males. In conclusion, this study established the standard as regarding the normal size of each lung lobe and lobule in relation to the age and sex of the RS goat. These parameters will aid diagnosis of lung disease and assessment of pulmonary consolidation at postmortem in a poor resource setting.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of Src Reduces Cortactin Phosphorylation and Transendothelial Migration of Acute Lymphoblastic Leukemia B cells
Authors: Camacho Karina Elizabeth Jimenez , Martinez Ramon Castellanos , Diaz Paola Briseño , Sevilla Jose Luis Flores , Schnoor Michael 
Published: June 2021

B‐cell acute lymphoblastic leukemia (B‐ALL) is the most common leukemia in children and adults. B‐ALL cells have the ability to extravasate and infiltrate bone marrow and organs such as lungs, testis and central nervous system (CNS). Dynamic actin cytoskeletal remodeling is needed to accomplish transendothelial migration. Cortactin is an actin‐binding protein which accumulates in lamellipodia and invadopodia and regulates neutrophil transendothelial migration during inflammation. Recently, we showed that cortactin is overexpressed in B‐cells of B‐ALL patients, and significantly correlates with bone marrow colonization, organ infiltration, drug resistance and disease relapse. Cortactin is a target for several posttranslational modifications such as phosphorylation and acetylation, being first discovered as a substrate of Src kinase and recently as substrate of the histone deacetylases SIRT1 and HDAC6. Phosphorylation of cortactin is thought to lead to lamellipodia and invadopodia protusion, whereas acetylation diminishes the affinity of cortactin to F‐actin, decreases cell migration capability, and may lead to nuclear translocation. However, it remains to be explored whether these modifications are required for the aggressiveness of leukemic B‐ALL cells. Thus, in this study, we analyzed the importance of such modifications for transendothelial migration and bone marrow organoid colonization. A MTT (3‐(4, 5‐dimethylthiazolyl‐2)‐2, 5‐diphenyltetrazolium bromide) assay was performed to determine effects of certain pharmacological inhibitors of posttranslational modifications on cell viability. To this end, REH cells were incubated with the indicated pharmacological compounds for 24 hours at different concentrations (50 to 150 mM). The IC50 value for PP2 (Src inhibitor) was 190.82 mM, for EX‐527 (SIRT1 inhibitor) was 190.94 mM, for Tubacin (HDAC6 inhibitor) was 41.39 mM, and for Tubastatin‐A (HDAC6 inhibitor) was 94.09 mM. Importantly, after 20 min of treatment of only REH cells with PP2 at 20 μM, REH cells transmigrated significantly less across CXCL12‐stimulated HUVEC monolayers, which may be due to a reduction of phosphorylated cortactin that impairs cytoskeletal remodeling needed for migration of the cells. Taken together, these results suggest that cortactin phosphorylation may be needed by leukemic B cells to achieve transmigration and may reveal cortactin as a potential new therapeutic target for high risk B‐ALL patients to prevent disease relapse.Support or Funding InformationCONACyTThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Association between Cadmium Air Exposure and Prostate Cancer Stage at Diagnosis
Authors: Vijayakumar Vish , Jagai Jyotsna S. , Abern Michael R. , Kajdacsy‐Balla Andre 
Published: June 2021

IntroductionThere is clear evidence that high levels of occupational exposure to cadmium (Cd) lead to a more aggressive prostate cancer (PC) tumor. At levels of exposure which are more relevant to the general population, there is disputing evidence as to whether Cd causes a PC tumor to become more aggressive. In this study, we assess the relationship between ambient exposure to air Cd and PC stage at diagnosis.MethodsWe collected outcome data from the Surveillance, Epidemiology, and End Results (SEER) for reported PC cases diagnosed from 2010 to 2014. PC stage at diagnosis was categorized as metastatic or localized. Exposure data was collected from the 2011 National Air Toxics Assessment (NATA) which was developed by the EPA to review air toxics in the United States. The total exposure concentration of Cd compounds in the air was a county‐level metric calculated by the EPA using weighted averages of air Cd concentration levels measured at various point emission sources. We calculated quintiles of Cd exposure concentration for ease of interpretation. Odds Ratios (OR) and 95% confidence intervals (CI) assessing the relationship between county‐level air Cd exposure and PC stage at diagnosis were calculated using multivariable logistic regression models. Analyses were adjusted for age at diagnosis, sociodemographic status, and overall air quality and were stratified by race.ResultsThe study cohort consisted of 234,991 PC cases from 594 counties reported by SEER. Of these cases, 92.5% were localized and 7.5% metastatic at diagnosis. Higher levels of air Cd exposure were associated with an increased likelihood of a PC case being metastatic at diagnosis. The adjusted odds ratios for the 80th percentile vs 20th percentile of Cd exposure were: all races (OR 1.030, CI 1.019 – 1.040), White (1.049, CI 1.037 – 1.062), Black (1.032, CI 1.0072 – 1.057), and other races (0.965, CI 0.925 – 1.0062).ConclusionHigher Cd air exposure concentration was associated with advanced PC stage at diagnosis with effects that vary somewhat by race. Since inhalation of Cd in the air only contributes to a small percentage of total Cd intake, further studies should be conducted to investigate the effects of other forms of Cd intake such as dietary and water exposures on PC aggressiveness.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Beta‐amyloid on cell death‐related molecules: Is the co‐chaperone BAG2 a potential target for Alzheimer's disease?
Authors: Carrettiero Daniel Carneiro , Fardin Nubia M , do Carmo Ramon Torreglosa , Almeida Maria Camila 
Published: June 2021

Alzheimer's disease is characterized by abnormal accumulation of extracellular beta‐amyloid plaques (Aβ). Aβ promotes cell death and NF‐κB signaling activation. Because BAG2 is repressed by NF‐κB signaling, the aim of the present study was to investigate the effect of Aβ/BAG2 on cell death‐related molecules such as NF‐κBp65, BAX, Bcl‐2, and Bcl‐3 in hippocampal primary cell culture and in differentiated and non‐differentiated SH‐SY5Y cells. In addition, we also investigate the effect of Aβ/BAG2 on NFκB1, NFκB2, RELA, and RELB gene expression. Western blotting and qPCR were used in the present project after Aβ stimulation with BAG2 overexpression or BAG2 knockdown.Aβ toxicity was increased in BAG2 overexpressing cells and was decreased after mRNA BAG2 knockdown. Aβ treatment promotes an increase in pNF‐kBp65/NF‐kBp65 suggesting nuclear translocation of this molecule and possible gene activation independently of BAG2. Pro‐apoptotic BAX and anti‐apoptotic Bcl‐2 proteins and mRNA were analyzed in Aβ‐stimulated cells with BAG2 knockdown. Aβ treatment resulted in an increase in BAX which was reversed in mRNA BAG2 knockdown cells, but no significant effect was detected in Bcl‐2 or Bcl‐3 levels. The effect of Aβ in differentiated compared to non‐differentiated SH‐SY5Y cells were distinct. Aβ treatment, in differentiated SH‐SY5Y cells, resulted in an increase of NFKB1, NFKB2, and RELA which was reversed in mRNA BAG2 knockdown cells. No significant effect was detected in RELB levels. On the other hand, in non‐differentiated SH‐SY5Y cells, Aβ treatment resulted in a decrease in NFκB1, NFκB2 and increase in RELB which was reversed in mRNA BAG2 knockdown cells.Our data suggests BAG2 an important molecule related to Aβ toxicity and NF‐κB signaling activation. BAG2 increased Aβ toxicity, probably interacting with the transcription factor NF‐κB in the NF‐κB nuclear translocation, and resulted in activation of pro‐apoptotic BAX leading to the expression of several cell death‐related molecules. Nevertheless, this effect seems to be differentially activated in differentiated and non‐differentiated SH‐SY5Y cells.Support or Funding InformationFAPESP (grant 2015/23426‐9)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Latent‐HIV‐1 Exosome Blockade Mitophagy Flux in Human Brain Microvascular Endothelial Cells
Authors: Chandra Partha K , Rutkai Ibolya , Kim Hogyoung , Braun Stephen E , Abdel‐Mageed Asim B , Mondal Debasis , Busija David W 
Published: June 2021

BackgroundInsufficient removal of dysfunctional mitochondria, due to a defective mitophagy pathway, leads to increased ROS, oxidative stress, and neuroinflammation which triggers the pathogenesis of various neurodegenerative diseases. HIV‐1‐associated neurocognitive disorders (HAND) is a common disease with variable reported prevalence in different populations (20– 69%). We investigated how exosomes from latent‐HIV‐1 infected cells impaired mitophagy flux in the human brain microvascular endothelial cells (HBMVEC).MethodsExosomes were isolated by ultracentrifugation from latent‐HIV‐1 infected T‐cells [exo‐J Lat(9.2)] and monocytes (exo‐U1) and characterized by the qNano‐IZON and NanoSight 300 systems by measuring the concentrations (3.2×1012 particles/mL), particle diameter (Mean: 100.5±28.8 nm), and size‐distribution (Mode: 75.5±3.5 nm) of exosomes from J Lat(9.2). To determine whether exposure to exo‐J Lat(9.2) could induce mitophagy, HBMVECs were exposed to varying doses of HIV‐exosomes (10, 25, 50 and 100 μg/mL) for 24 and 48 h, after which the expression level of mitophagy markers PINK1, Parkin, and Drp1 were determined using western blotting. Carbonyl cyanide 4‐(trifluoromethoxy) phenylhydrazone (FCCP; 5 μM for 3 h), a known mitophagy inducer, was used as a positive control.ResultsMitophagy is a two‐step process by which damaged mitochondria are first primed by PINK1/Parkin proteins and then eliminated via the autophagic machinery. We then examined the expression levels of autophagy (macroautophagy) proteins: the autophagosome initiation marker (Beclin 1), autophagosome formation marker (LC3B‐II), and autophagy degradation marker (SQSTM1 or p62) in HBMVECs exposed to latent‐HIV‐1‐exosomes. Interestingly, following exposure of HBMVECs to exo‐J Lat(9.2), the expression level of PINK1/Parkin proteins were upregulated in a dose dependent manner, with a concomitant increase in the formation of autophagosomes, shown by increased expression of Beclin 1 and LC3B‐II. FCCP treated cells showed decreased expression of p62 indicated by the active presence of mitophagy flux. Remarkably, cell exposure to HIV‐exosome resulted in increased expression of p62, indicating a possible blockage of the mitophagy flux, leading to the accumulation of mitophagosomes.ConclusionsOur preliminary study indicated that latent‐HIV‐1 exosome increased mitochondrial damage via defective mitophagy.Support or Funding InformationGrant Numbers: U54 GM104940, AHA 17SDG33410366, HL‐093554.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Serum Estradiol/Testosterone Ratio Affects the Development of Benign Prostate Hyperplasia in Rats
Authors: Liu Xiangyun , Li Kai , Liu Zhiwei , Gui Shukang 
Published: June 2021

ObjectiveBenign prostatic hyperplasia (BPH) develops more likely with increasing age and changing serum concentrations of circulating estradiol (E2) and/or testosterone (T).MethodsIn this study we explored the relationship between serum E2/T ratio and BPH risk in rats by fitting a mathematical model. A total of 176 male rats were randomized divided into normal control, castrated control, and 20 more groups of castrated animals were administered subcutaneously once daily with one of the four dose levels (0.06, 0.17, 0.5, and 1.5 mg per rat) of T combined with one of the five dose levels (2, 6, 17, 50, and 150μg/kg) of E2 in olive oil by different dose ratio. After 30 days, Serial blood samples were obtained to determine serum T and E2 levels by magnetic bead enzyme‐linked immunosorbent assay. The volume of prostate was measured by water displacement method. The prostate specimens were then fixed, embedded in paraffin, and subjected to hematoxylin and eosin staining for pathological examination. MATLAB software was used to simulate the relationship between prostate/body weight ratio (PBR) and E2/T ratio with a mathematical equation.ResultsCompared with the normal control group, the prostate volume decreased significantly (P < 0.05) in the castrated control group and the treatment groups receiving E2/T dose of 2/0.06 or 6/0.06, while the prostate volume increased significantly (P < 0.05) in the other treatment groups. Compared with the normal control group, the PBR increased statistically significant in the treatment groups receiving E2/T dose of 50/0.5, 150/0.5, 2/1.5, 6/1.5,17/1.5, 50/1.5, or 150/1.5 (P < 0.05). The values of PBR, E2 and T in the treatment groups were significantly higher than those in the control groups. Stepwise regression showed that PBR was a function of E2 and T. PBR=−0.1782 + 0.0081E2 + 0.063T − 0.6 × 10−5E2 2 − 0.28 × 10−3T2, (R2=0.6317)ConclusionE2 /T ratio change may be one of the risk factors for PBR, which is associated with the development of BPH.Support or Funding InformationNational Natural Science Foundation of China (No.: 81772429)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Low‐intensity ultrasound differentially upregulates cell death regulators in cancer cells, and downregulates inflammatory pathways in non‐cancer cells via oxidative stresschromatin long‐range interaction signaling
Authors: Yang Qiao , Wang Jiwei , Lai Bin , Sun Yu , Yu Daohai , Yang William , Shao Ying , Zhang Chunquan , Liu Yanna , Wang Hong , Yang Xiaofeng 
Published: June 2021

The mechanisms underlying low intensity ultrasound (LIUS) suppression of inflammations and tumorigenesis remain poorly determined. Here, we determined the mRNA expressions of 299 cell death regulators in 13 cell death types in the microarray data. We made the following findings: 1) LIUS upregulates more cell death regulators in cancer cells than in non‐cancer cells, and downregulates more inflammatory regulators in non‐cancer cells than cancer cells; 2) LIUS‐induced most top signaling pathways in cancer cells are not shared with that in non‐cancer cells. LIUS induces more cell death related pathways in cancer cells but inhibits the expressions of a list of inflammation‐related regulators via TP53‐, and SRF‐, mediated pathways, respectively, in non‐cancer cells; 3) LIUS shares 10–25% of the top pathways with mild hyperthermia in regulating cell death regulator expression regardless of cell types; and shares 15–45% of the top pathways with oscillatory shear stress (OSS) in regulating gene expressions in non‐cancer cells; and 4) LIUS may modulate chromatin long range interactions to regulate gene expressions in cells; upstream chromatin long‐range interaction sites (CLRISs) are more favorable than downstream CLRISs for LIUS modulation of gene expressions in cancer cells; and downstream CLRISs play more important roles than upstream CLRISs for LIUS modulation of gene expressions in non‐cancer cells. Our finding demonstrated that LIUS suppresses tumorigenesis and inhibits inflammation by modulating the expressions of cell death regulators, which leads to novel therapeutic targets for treating cancers and inflammation.Support or Funding InformationThis work was supported by NIH grant to Drs. XF. Yang, H. Wang.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Modulatory Effects of Cannabidiol on Microglial Function and Receptor Expression in Epilepsy
Authors: Victor Tanya R. , Nissen Jillian C. , Elmes Matthew W. , Deutsch Dale G. , Tsirka Stella E. 
Published: June 2021

Epilepsy, a chronic disorder characterized by abnormal brain cell activity leading to recurrent seizures, has recently seen increased and FDA‐approved usage of cannabidiol (CBD) as a treatment for intractable epilepsies. CBD, a non‐psychotropic component of Cannabis sativa, is reported to lessen the severity of experimentally induced seizures in animals and to suppress neuroinflammation in culture. This compound has been able to provide therapeutic options for the nearly 30% of patients who do not respond to traditional epilepsy medications. Although CBD has been shown to increase intracellular levels of anandamide, an endogenous cannabinoid, its mechanism of action is still poorly understood. Our lab has shown that microglia, the immune cells of the central nervous system (CNS), act as important mediators of seizure severity. The ability of microglia to modulate seizures has been associated with the activation of Toll‐like receptors (TLRs), which play important roles in pathogen recognition and inflammation. Microglia also contribute to neurogenic processes and synapse maturation, aiding in the formation of functional neuronal circuits. Our preliminary data suggest that CBD functions partially through microglia. We postulate that CBD modulates the activation of microglial cells in the brain during experimental epilepsy. In this study, we evaluate the effects of CBD on cell death, inflammation and microglial receptor expression in vitro and in vivo post seizure.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.