bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Oncogene MYC Regulates the Branched Chain Amino Acid Metabolism and mTOR Signaling in Diffuse Large B Cell Lymphoma
Authors: Follett Aleyna , Figueroa Lucas , Boyer Michael , Ananieva Elitsa 
Published: May 2022

Diffuse large B‐cell Lymphoma (DLBCL) is the most common type of aggressive non‐Hodgkin lymphoma (NHL) worldwide. While therapeutic strategies to combat DLBCL have advanced, these malignancies continue to be challenging to treat due to their heterogeneity and frequently recurring mutations. Aberrations in the oncogene MYC are commonly found in high grade DLBCL and are associated with poor responses to standard therapy.In search of downstream targets of MYC in DLBCL, we found MYC‐binding sites in the promoter regions of BCAT1, BCAT2, and BCKDHA. These genes encode enzymes in the metabolism of the branched chain amino acids (BCAAs), and they have been previously linked to cancer progression. BCAT1 and 2 encode the cytosolic (BCATc) and mitochondrial (BCATm) branched‐chain aminotransferases, while BCKDHA encodes the E1α subunit of the branched‐chain ketoacid dehydrogenase complex. We hypothesized that MYC activates these genes in DLBCL tissues leading to upregulation of BCAA metabolism and lymphoma growth.Human tissues collected from DLBCL patients (n=89) or controls (n=33) were used to analyze the gene expression of MYC, BCAT1, BCAT2 and BCKDHA followed by correlative analysis of the expression of these genes and the survival of DLBCL patients. Next, human DLBCL and B‐lymphoblast (control) cells were treated with the MYC inhibitor 10058‐F4 (0, 25, 50, 100 µM), the BCAA antagonist N‐acetyl leucine amide (NALA, 0, 10, 20, 40 mM) and the BCATc inhibitor gabapentin (0, 5, 10, 20 mM). 24 hours post treatment, the cell viability and cell proliferation, as well as the protein expression of BCATc, BCATm, and E1α, and the total and phosphorylation states of the ribosomal protein S6 (a downstream target of the mammalian target of rapamycin, mTOR) were determined.Results showed that the gene expression of BCAT1, BCAT2and BCKDHA were significantly elevated in tumor tissues from DLBCL patients compared to controls, and there was a positive correlation between their expression and the upregulation of MYC in DLBCL tissues. The high expression of MYC and BCAT2, but not that of BCAT1 and BCKDHA, correlated with poor overall cancer survival. Inhibiting the MYC function by 10058‐F4, the BCAA uptake by NALA, and the BCATc activity by gabapentin led to concentration dependent reductions in the cell viability and proliferation of the DLBCL cells. The protein expression of BCATm and E1α were reduced in the presence of 10058‐F4, which correlated with downregulation of mTOR signaling as measured by the deceased phosphorylation state of S6 in treated DLBCL cells. These results support the hypothesis that MYC is an upstream regulator of genes associated with BCAA metabolism. The results also demonstrate that DLBCL cells need MYC and BCAAs to upregulate mTOR signaling and to survive and may represent a new molecular mechanism via which MYC exerts its effects in DLBCL lymphoma.
Pentadecapeptide BPC 157 Counteracts Hemodynamic and ECG Disturbances Induced by Sotalol in Rats
Authors: Premuzic Mestrovic Ivica , Smoday Ivan Maria , Mestrovic Tomislav , Zizek Helena , Skrtic Anita , Skorak Ivan , Haluzan Damir , Figl Josip , Brizic Ivan , Staresinic Mario , Pavic Predrag , Seiwerth Sven , Sikiric Predrag 
Published: May 2022

Cytoprotective and organoprotective pentadecapeptide BPC 157 demonstrated particular beneficial cardiovascular effects (Curr Pharm Des. 2014;20(7):1121‐1125). Antiarrhythmic effects counteracted digoxin toxicity (Regul Pept. 2009;156(1‐3):83‐89), major blood vessel occlusions (Biomedicines 2021, 9,1029; 2021, 9,609; 2021, 9, 792) and lidocaine toxicity (Emerg Med Int. 2020;2020:6805354). Now, we reveal, by BPC 157, mitigation of hemodynamic and ECG disturbances of sotalol, a class III antiarrhythmic with non‐selective beta‐blocking activity.Albino Wistar rats received sotalol (80mg/kg, intragastrically), and at 5 min thereafter were treated with saline or BPC 157 (10ng/kg, intragastrically. Seven standard and one precordial ECG leads were recorded (baseline; after application of sotalol; and 10, 30, 90, 120 and 150 minutes after application of sotalol). Heart rate, QRS complex, PR and QT intervals were analyzed. Corrected QT intervals (QTc) were calculated according to Fridericia formula.Baseline ECG readings included average heart rate of 275.5 ± 25 bpm, average PR 54.8 ± 3.4 ms, average QRS complex 25.0 ± 4.23 ms and average QTc 166 ± 9.7 ms. Sotalol caused bradycardia (245 ± 23.5 bpm), PR prolongation (57.3 ± 2.7 ms), QRS widening (26.3 ± 3.2 ms), and QTc prolongation (173 ± 9.1 ms), measured 10 minutes after application. In BPC 157 treated animals (5 minutes after BPC‐157 application, 10 minutes after sotalol application), these effects were attenuated (heart rate 306 ± 31.7 bpm, average PR 52.3 ± 1.6 ms, average QRS 23.7 ± 5.4 ms and average QTc 159 ± 21.2 ms). This attenuation was lost in later measurements, for QRS ‐ 25 minutes after BPC 157 application, for frequency after 55 minutes, for PR after 115 minutes and for QTc after 145 minutes. In animals treated with sotalol, an increase in superior sagittal sinus pressure (10 ± 5 mmHg), as well as an increase in portal vein (17 ± 4 mmHg) and inferior caval vein pressures (12 ± 6 mmHg) were noted, along with the decrease in abdominal aortic pressure (53 ± 10 mmHg). These effects were attenuated in BPC treated animals (‐1 ± 1 mmHg for superior sagittal sinus, 7 ± 1 mmHg in portal vein, 5 ± 1 mmHg in inferior caval vein, and 81 ± 7 mmHg in abdominal aorta).In conclusion, BPC 157 attenuated deleterious hemodynamic effects of sotalol while the effects on the ECG parameters were also noted, although not long lasting, supporting the view of BPC‐157 as a rapid‐acting protective agent.
Monensin and its analogues show anti‐glioblastoma activity in an organoid model of cancer
Authors: Urbaniak Alicja , Reed Megan R. , Heflin Billie , Gaydos John , Piña‐Oviedo Sergio , Jędrzejczyk Marta , Klejborowska Greta , Stępczyńska Natalia , Chambers Timothy C. , Tackett Alan J. , Rodriguez Analiz , Huczyński Adam , Eoff Robert L. , MacNicol Angus M. 
Published: May 2022

Glioblastoma (GBM) is the most common primary malignant brain tumor in adults. Despite numerous clinical trials, the standard of care therapy has remained unchanged for the last decade. One of the main problems contributing to the limited effective treatment options and poor overall survival is the lack of models which can reliably recapitulate tumor heterogeneity. Organoids are 3D self‐organized structures which mimic tumor architecture, microenvironment, and cellular interactions which makes them an improved model for anti‐cancer drug discovery.Monensin (MON) is a polyether ionophore antibiotic characterized by wide range of biological properties including anti‐cancer activity. In order to identify more potent compounds based on the scaffold of MON, we investigated the anti‐GBM activity of 14 novel esters and urethanes of MON. In 3D mini‐ring cell viability assays, we identified seven analogues (IC50 = 91.5 ± 54.4 ‐ 291.7 ± 68.8 nM) more potent towards GBM than the parental MON (IC50 = 612.6 ± 184.4 nM). These analogues induced DNA fragmentation in an organoid model of GBM, suggestive of apoptotic cell death. Furthermore, the most potent analog, compound 1, significantly reduced GBM cell migration, induced PARP cleavage and degradation, increased ɣH2AX signaling and increased expression of the autophagy marker LCII. To investigate the activity of these novel compounds in a tumor microenvironment, we have developed a host:tumor hybrid 3D organoid system. For host tissue, we generated human cerebral organoids (COs) from hiPSCs. The COs displayed multiple neural rosettes with a proliferative zone of neural stem cells (Nestin+), neurons (TUJ1+), primitive ventricular system (SOX2+/Ki67+), intermediate zone (TBR2+) and cortical plate (MAP2+). These findings suggest the level of differentiation and development of our COs as equivalent to early stage human fetal brain. We then co‐cultured RFP‐labeled U87MG cells with fully formed COs. After establishing U87MG tumor formation, hybrid organoids were treated with MON or compound 1. Compound 1 significantly reduced U87MG tumor size after four days of treatment. Our findings highlight the therapeutic potential of MON analogues towards GBM and support further research and clinical development of these compounds.
The Impact of Secondary Bacterial Infections in COVID‐19 Patients on Mortality and the Immune Response in the State of Arizona
Authors: Potter Pamela , Jiaxin Bai , Villait Akash , Khin Brian , Vu Cathy , Ihms Benjamin , Krzeczowski Rachel , Ashurst John , Santarelli Anthony , Smith Victoria 
Published: May 2022

The literature on COVID‐19 has focused primarily on urban centers and suggest an increased risk of mortality in COVID‐19 patients coinfected with bacteria compared to those solely infected with COVID‐19. In patients screened or admitted to suburban and rural hospitals in Arizona, mortality increased with co‐infection. The impact of bacterial coinfections on the immune response was studied by comparing white blood cell (WBC) and lymphocyte counts as well as the relationship between duration of steroid use and mortality in coinfected COVID‐19 patients investigated.MethodsA chart review of 677 SARS‐CoV‐2 patients was conducted at during December 2020 to March 2021. Patients solely infected with COVID‐19 (COVID‐19 patients) were compared to patients with concurrent bacterial or fungal infections (coinfected patients). Parameters included admission duration, ICU duration, mortality, WBC, lymphocytes, and duration of steroid use. WBC and lymphocytes were compared amongst COVID‐19 and coinfected patients that survived or that expired during their hospital stay. COVID‐19 positive patients not admitted to hospital were used as controls.ResultsThe admitted and ICU duration of coinfected patients was significantly (p < 0.05) higher (12.93 vs 10.92 days) than COVID‐19 patients (6.39 vs 4.38 days), as was mortality rate (22.5% vs 9.8%). WBC was higher (9.4x103) in coinfected patients than COVID‐19 alone (7.9x103). Lymphocytes were lower (2.03x103) in coinfected patients compared to COVID‐19 alone (2.41x103). A significant difference in WBC was seen between expired COVID‐19 patients (10.1x103), expired coinfected patients (10.01x103) and COVID‐19 non‐admitted patients (6.44x103). Lymphocytes were lower in surviving coinfected patients (I.96x103) and expired COVID‐19 patients (I.61x103) than COVID‐19 non‐admitted patients (7.33x 103). Expired coinfected patients were treated with steroids for a longer average duration (9.22 days) when compared to expired COVID‐19 (6.77 days).ConclusionThe presence of other bacterial infections increased hospitalization and mortality rate of patients admitted to a suburban hospital with COVID‐19 infection. Patients with co‐infections needed longer steroid treatment, had higher WBCs but lower lymphocytes. Thus, co‐infection presents significant challenges for treatment of patients with a diagnosis of COVID‐19.
Differences in Immune Cell Mitochondrial Function in Black and White Patients with Heart Failure with Preserved Ejection Fraction
Authors: Bohmke Natalie J. , Chavez Domenico A. , Billingsley Hayley E. , Damonte Juan I. , Del Buono Marco G. , Robinson Austin T. , Markley Roshanak , Carbone Salvatore , Abbate Antonio , Kirkman Danielle L. 
Published: May 2022

BACKGROUNDHeart failure with preserved ejection fraction (HFpEF) affects patients of all races and ethnicities. However, Black/African American patients are historically underrepresented in clinical research of HFpEF and have been reported to have unfavorable outcomes, yet the underlying mechanisms are not entirely understood. Excess proinflammatory cytokine and reactive oxygen species production from immune cells contributes to increased cardiovascular risk and may be a cause or consequence of alterations in immune cell metabolism. The aim of this study was to describe the immune cell bioenergetic profile in Black and White patients with HFpEF.METHODSWe studied 20 patients with HFpEF. 13 patients self‐reported as Black or African‐American (11 female; Median [IQR]: Age: 55 [50, 66] yrs; BMI 39 [36, 43] kg/m2; hsCRP 5.3 [3.5, 9.3] mg/dL; BNP 50 [50, 200] pg/mL) and 7 self‐reported as White (7 female; Age: 68 [43, 72] yrs; BMI 34 [33, 40] kg/m2; hsCRP 3 [1, 4.9] mg/dL; BNP 109 [50, 168] pg/mL). The mitochondrial bioenergetic profile was assessed in isolated peripheral blood mononuclear cells by a Seahorse XFp Analyzer. Respirometry coupled with modulators of cellular respiration were used to obtain basal respiration, ATP‐linked respiration, maximal respiration, proton linked respiration, spare respiratory capacity, and non‐mitochondrial respiration. The Mann Whitney U test was used to compare variables between groups.RESULTSProton leak respiration (Median [IQR]: 29 [4, 66] vs. 2.5 [1, 3] pmol/min, p=0.013), basal respiration 93 [60, 138] vs. 57 [53, 69] pmol/min, p = 0.008), maximal respiration (323 [191, 432] vs. 185 [162, 238] pmol/min, p = 0.0370) were higher in Black versus White patients. Raw spare respiratory capacity (232 [128, 293] vs. 125 [105, 145] pmol/min, p = 0.037) was also higher in Black versus White patients, whereas relative spare respiratory capacity (293% [276, 378] vs. 307% [259, 357] pmol/min, p=0.87) was not different between groups. There was a trend for lower coupling efficiency (94% [55, 95] vs. 97% [94, 99] pmol/min, p = 0.08) in Black vs White patients.CONCLUSIONIn an unselected cohort of patients with HFpEF, immune cell mitochondria basal respiration and respiratory reserve, as indicated by maximal respiration and spare respiratory capacity, are higher in Black versus White patients. Future studies are warranted to better understand the potential pathophysiological consequences of the observed differences.
Understanding Molecular Heterogeneity in Hepatocellular Carcinoma
Authors: Delgado Evan R. , Tao Junyan , Bell Aaron W. , Hu Shikai , Poddar Minakshi , Singh Sucha , Monga Satdarshan P. 
Published: May 2022

Hepatocellular carcinoma (HCC) is the 5th most common cause of cancer‐related death in the United States with an estimated 32,000 annual deaths. Roughly 30% of HCC cases demonstrate mutations in the CTNNB1 oncogene that encodes a form of the ꞵ‐catenin protein that cannot be degraded. Recently, we developed a mouse model of HCC that genetically represents ~9‐12% of all human HCCs. Here, we combine hydrodynamic tail vein injections with the Sleeping Beauty Transposase – a non‐viral method of expressing genes of interest. Our model uses a clinically observed mutant of ꞵ‐catenin‐(T41A) that is co‐expressed with a mutant form of the nuclear factor erythroid 2–related factor 2 (Nrf2‐G31A) in the livers of wild‐type mice (B+N). This system results in microscopic HCC within 3 weeks and evident macroscopic disease by 7 weeks after induction.ꞵ‐catenin activation can be identified by the expression of its downstream target gene Glutamine Synthetase (GS) in the liver. Patient HCCs with ꞵ‐catenin mutations strongly correlate with GS expression. The evolution and function of GS in b‐catenin‐mutated HCC, remains unclear. Also, there is a gap in the knowledge whether elevated GS expression correlates with HCC biology, which were all subjects of the current study. B+N mouse model of HCC was examined temporally for development of HCC along with GS expression. B+N mice develop HCCs with heterogeneous GS expression in the form of GS‐high and GS‐low to GS‐negative foci. We next asked whether GS expression is associated with cell proliferation or death in the B+N model. We stained tissues for TUNEL and PCNA to mark cell apoptosis and proliferation, respectively in 7‐ and 9.5 weeks post‐injection livers, which were then correlated to GS staining of HCC nodules. While we found no correlation between apoptosis and GS, we found nodules with lower GS expression to have higher proliferation and vice versa. Finally, we asked if eliminating GS expression from B+N HCCs will impact tumor burden. Five weeks after B+N tumor induction in GS‐floxed mice, these animals were injected with AAV8‐TBG‐Cre virus to delete Glul,and mice were harvested after 2‐ or 4.5‐weeks. We found B+N mice with Glul deletion show increased liver weight to body weight ratio, a measurement of HCC burden, compared to controls. The livers of these mice were enriched for GS‐negative tumors. Together, our data suggests that HCC nodules with lower GS expression may outperform GS‐positive foci, and thus are more aggressive compared to the nodules with higher GS expression, which may be more differentiated. The mechanisms of GS heterogeneity and elucidation of its functional significance may help select the most effective targeted therapies for HCC patients with ꞵ‐catenin mutations.
The Effects of Elevated Sodium on Mitochondrial Function in Peripheral Blood Mononuclear Cells
Authors: Mehrer Justin , Patik Jordan C. , Shenouda Ninette , Edwards David G. 
Published: May 2022

INTRODUCTIONDiets high in sodium are associated with impaired vascular function independent of blood pressure. Recent data suggests that sodium accumulation from high salt diets reduces mitochondrial respiration in monocytes and macrophages, resembling what occurs during an inflammatory response. A similar response in circulating peripheral blood mononuclear cells (PBMCs) could implicate systemic inflammation as a mechanism contributing to high salt‐induced vascular dysfunction. Therefore, we aimed to test the hypothesis that exposure to a high sodium environment reduces PBMC mitochondrial respiration.METHODSPBMCs were separated from whole blood of ten healthy subjects (7 men/3 women, 18‐30 years, blood pressure ≤ 130/80mmHg; BMI < 30 kg/m2). PBMCs were then seeded at 2x105 cells/well in Poly‐D‐lysine treated cell culture plates. Plated cells were incubated for 24 hours in RPMI 1640 medium with 10% FBS at 137.7 mM sodium (normal sodium, NS) and 180 mM sodium (high sodium, HS). Basal, maximal, and ATP‐linked oxygen consumption rates were assessed post‐incubation by respirometry assay via Seahorse XFp analyzer (Agilent). Spare respiratory capacity was calculated as the difference between maximal and basal oxygen consumption rate.RESULTSBasal oxygen consumption rate was significantly reduced in HS compared to NS (65.0 ± 12.7 vs. 39.6 ± 6.5 pmol/min, p = 0.03), as was maximal oxygen consumption rate (198.3 ± 32.6 vs. 148.4 ± 27.2 pmol/min, p = 0.008). Basal ATP‐linked oxygen consumption rate was also significantly reduced in HS compared to NS (57.3 ± 9.8 vs. 34.4 ± 5.4 pmol/min, p = 0.01). Spare respiratory capacity in HS was significantly reduced compared to NS (133.2 ± 23.4 vs. 108.8 ± 21.2 pmol/min, p = 0.03).CONCLUSIONReduced basal and maximal oxygen consumption rates and spare respiratory capacity provide evidence that elevated sodium exposure reduces PBMC mitochondrial respiration. Additionally, high sodium exposure reduced ATP‐linked respiration potentially indicating alterations in PBMC energetics, resembling what occurs during inflammation. Collectively, our preliminary findings warrant further investigation into the role of sodium induced mitochondrial dysfunction in PBMCs as a potential mechanism contributing to high salt‐induced vascular dysfunction.
The potential role of microRNA as bio markers in canine congestive heart failure
Authors: Mitchell Carol L. 
Published: May 2022

ObjectiveIn our focus regarding microRNA, single‐stranded, small non‐coding RNAs, we aim to understand miRNA's roll and functional importance concerning congestive heart failure (CHF) and myxomatous mitral valve degeneration (MMVD). Characterizing expression profiles of circulating microRNAs via genome‐wide sequencing for dogs with CHF secondary to MMVD.Animals8 healthy client‐owned dogs and 8 middle age‐matched client‐owned dogs with CHF secondary to MMVD.ProceduresBlood samples were collected before administering cardiac medications for the management of CHF. Quantitative reverse transcription PCR (qRT‐PCR) assays were used to validate expression profiles of differentially expressed circulating microRNAs identified from NGS analysis of dogs with CHF.Conclusion and Clinical RelevanceAwaiting data indicating the patterns of microRNA expression are distinctive and denote molecular biomarkers with CHF in dogs with MMV.
Myostatin Deletion Protects Against Renal Dysfunction During a High Salt Diet
Authors: Wright Carson L. , Nunan Emily L. , Butcher Joshua T. 
Published: May 2022

Chronic Kidney Disease (CKD) afflicts ~15% of U.S. adults and occurs when the kidneys are ineffective in filtering the blood of waste products. CKD is irreversible and current interventions are focused on protecting and preserving renal function. Risk factors for CKD are multifactorial but growing evidence suggest that a key risk factor is dietary salt intake. A diet high in sodium will increase fluid retention, blood pressure, proteinuria, and ultimately drive the progression of kidney damage. Currently, it is estimated that 90% of the U.S. population consumes excess dietary sodium. As such, it is a medical necessity that interventions focus on ways to optimize renal sodium excretion and protect kidney function over time.Exercise is a key therapy to improve kidney function but a large amount of people are unable to exercise due to disability, lack of time or money, or a pandemic. Our lab has previously discovered that when myostatin, a negative regulator of skeletal muscle mass, is deleted from lean mice it also improves kidney function in diabetes. However, it is unknown whether myostatin deletion will also improve renal sodium handling. Thus, the hypothesis is that myostatin deletion improves sodium handling and protects against renal damage during a high salt diet.The experiment used adult lean mice with and without myostatin constitutively deleted. Both groups of mice were placed on a high salt (HS) diet (4% NaCl) for 14 days. Food consumption, water intake, and urine production was assessed at baseline, acutely (first 3 days on HS diet) and at the end of the experiment using metabolic cages. Blood pressure studies (in vivo) are ongoing and markers of renal (dys)function are currently being assessed, along plasma hormones that regulate fluid balance. Significance was determined at P<0.05.The results showed that myostatin deletion protects against kidney dysfunction during a high salt diet, as the myostatin KO mice consumed significantly more food, but maintained fluid balance (drank and urinated similar amounts to the lean control). Blood pressure remained lower in the myostatin KO compared to control. Importantly, myostatin deletion significantly blunted renal hypertrophy (68% less mass) compared to the control. Taken together, myostatin deletion improves the efficiency of the kidney in handling sodium and maintains fluid balance during a high salt diet. Thus, pharmaceutical inhibition of myostatin may prove an effective target for long‐term protection against high salt diets and preserving kidney function during CKD.