bluuebunny/crossref_metadata_2025 huggingface dataset:
Declining Attention Span in Medical Students: Fact or Fiction?
Authors: Ettarh Raj , Al‐Hussaini Heba , Hassan Sherif , Kilarkaje Narayana
Published: June 2021

Background

Despite a lack of previous investigations, there is a common consensus that attention span among young learners is declining. The millennial generation is described as having preferences for teamwork, interactive learning experiences, and visual media but suffer from short attention spans. To address the lack of empirical data, this study examined self‐reported attention span in medical students at matriculation. Only results relating to age and self‐reported attention span are presented here.

Methods

Two groups of matriculating medical students were surveyed by questionnaire shortly after starting in medical school: 334 first‐year medical students at Tulane University (who are required to graduate with a BS degree before entry into medical school), and 108 first‐year medical students at Kuwait University (who are required to graduate high school before entry into medical school). They were asked to respond to questions relating to learning habits and preferences.

Results

The age range for Tulane University medical students was 20–54 years with a mean of 24.3 ± 3.0 SD. The age range for Kuwait University medical students was 17–21 years with a mean of 18.8 ± 0.7 SD. Due to the wide range of ages, only data for mean age ± 1SD for both cohorts were analyzed. The average self‐reported attention span for Tulane University medical students was 69.4 ± 46.4 minutes compared to 59.3 ± 51.8 minutes for Kuwait University medical students. Median attention span was shorter in the Kuwait cohort than in the Tulane cohort. Average attention span for women in the Kuwait cohort was shorter than in the Tulane cohort, but shorter among men in the Tulane cohort than in the Kuwait cohort.

Conclusion

The self‐reported attention span was lower in the younger medical students at Kuwait University. The data suggest that instructional activities, especially lectures, that last less than 60 minutes may hold the attention of older medical students, but less so for younger medical students.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sorafenib supresses TGF‐β responsiveness by promoting TGF‐β type II receptor degradation.
Authors: Wang Shi‐Weigh
Published: June 2021

Transforming growth factor‐beta (TGF‐β) family are multifunctional and essential in growth and development. The multi‐kinase inhibitor sorafenib is the FDA approved drug for the treatment of many solid tumor. Previous studies have showed that sorafenib inhibit TGF‐β induced epithelial‐mesenchymal transition (EMT) and tissue fibrosis; but the detailed mechanism remains unknown. In this study, we found that sorafenib specific decreases TGF‐β type II receptor in liver epithelial cell through lipid raft/caveolae‐mediated endocytosis but not in hepatic stellate cell. We next find more detail mechanism to prove this different effect between epithelial and stellate cell. The degradation of TGF‐β type II receptor lead to suppression of TGF‐β signaling. Since TGF‐β have been reported that play important roles in several diseases, our study may provide a potential therapy of TGF‐β related disease.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Synthesis of Aspirin Analogs for Anticancer and Antibacterial Testing
Authors: Sleda Melissa A. , Albasrawi Hadeel K. , Timmons Shannon C.
Published: June 2021

Aspirin is a common over‐the‐counter drug used to treat pain, fever, and inflammation. It is one of the most widely used medications in the world with an estimated 35,000 metric tons produced and consumed annually. Recent research has shown that the regular ingestion of aspirin reduces a person's risk of developing colon cancer and even promotes colorectal tumor regression. Moreover, aspirin has a demonstrated ability to inhibit the proliferation of colorectal cancer cells in vitro. Although the mechanism of action has not yet been unequivocally established, it is clear that the synthesis of novel aspirin analogs is of medicinal interest. In this study, a library of aspirin analogs of varying degrees of hydrophobicity and steric hindrance were synthesized using salicylic acid as the molecular scaffold and a variety of acyl chlorides. These aspirin analogs will be screened against an array of cancer cell lines and will also be evaluated as antibacterial agents to determine their medicinal activity. The medicinal testing of this library of aspirin analogs will help to establish new structure‐activity relationships.

Support or Funding Information

We would like to thank the Department of Natural Sciences at Lawrence Technological University and the Howard Hughes Medical Institute Inclusive Excellence grant for support and funding.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Advancing Professional Development Strategies for Undergraduates in Chemistry and Biochemistry
Authors: Brown Christian , Fallucca Amber , Makris Thomas
Published: June 2021

In the Department of Chemistry and Biochemistry at the University of South Carolina (the Chemistry Department), most curriculum‐driven professional development for undergraduates is reserved for third‐ and fourth‐year students. Academic instruction occurs at all levels, but the context comes late. Early exposure to career options and hands‐on training is imperative for undergraduates to obtain maximum experiential education. The purpose of this project is to advance the current professional development strategies of the Chemistry Department, specifically targeting second‐ and third‐year students. The study will survey students' current involvement in professional development activities as well as how the Chemistry Department is facilitating undergraduate involvement in professional development. We will use the data on extracurricular experience and post‐graduation interest to design a workshop series to reach students with practical tools for success, such as networking skills and synthesizing intra‐ and extracurricular experiences. The impact of each workshop will be assessed with post‐session surveys asking participants about the value and applicability of the information shared. The most beneficial topics will be incorporated into curriculum development of the Chemistry Department's one‐credit hour undergraduate seminar course. By integrating more advanced professional development into the curriculum, chemistry and biochemistry undergraduates will be encouraged to pursue experiential education in their second and third years.

Support or Funding Information

This work was supported in part by the South Carolina Honors College Senior Thesis/Project Grant (CAB) and a grant from the University of South Carolina Magellan Scholar Program.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Use of Biometrics to Determine Differences in How Biochemistry Experts and Novices Read Metabolic Pathways
Authors: Cortes Kimberly Linenberger , Kammerdiener Kimberly , Randolph Adriane
Published: June 2021

Being able to move between multiple representations is an important skill in biochemistry. Schematics are a type of representation commonly used in biochemistry instruction often combined with chemical structures to form a metabolic pathway. As little work has focused on schematics in biochemistry, this study uses the Tobii X2‐30 Compact Eye Tracker system to record biochemistry experts and students enrolled in a one semester biochemistry course and a second semester organic course as they read a familiar (glycolytic) and novel (IMP synthesis) metabolic pathway image. This presentation will focus on differences in areas of interest (AOI), fixation, and scan path analysis.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sub‐Chronic Exposure to Organic Arsenic Disrupts Hepatic Energy Metabolism and Antioxidant System
Authors: Babayemi David Oluwagbemi , Adeyi Olubisi Esther , Dosumu Adebisi , Adubiagbe Hassan Taiwo , Abdulsalaam Mariam Temitope , Tijani Aliu , Olaleye Fausat Morenikeji , Akinhanmi Temilade Fola , Ugbaja Regina Ngozi , Ademuyiwa Oladipo
Published: June 2021

Arsenic is a ubiquitous toxicant, having a wide range of toxic effects on different systems in the body. To investigate the sub‐chronic effects of exposure to low doses of cacodylic acid (organic arsenic) on some enzymes of energy metabolism and antioxidant system in the liver, rats were exposed to cacodylic acid (20 ppm and 40 ppm) in their drinking water for ten weeks. Control animals received distilled water. Activities of catalase (CAT), glutathione peroxidase (GPx), glutathione‐S‐transferase (GST), superoxide dismutase (SOD), NAD glycohydrolase (NADase), xanthine oxidase (XO) and lactate dehydrogenase (LDH) and the levels of nitric oxide (NO), reduced glutathione (GSH) and lipid peroxidation (MDA) were determined in hepatic cytosol while malate dehydrogenase (MDH) and respiratory chain enzymes (complexes I, II and IV) activities were determined in the mitochondrial fraction. There were significant (p<0.05) decreases in the level of GSH and activities of NADase, MDH and complex IV in the exposed groups compared with the control. In the 20 ppm group, activity of hepatic SOD and level of NO increased by 419% and 142% respectively. A hormetic response was observed in the activity of GST. The activities of complexes I and II decreased as a result of exposure to cacodylic acid, these decreases were dose‐dependent. Our findings indicate that disruption of energy metabolism and antioxidant system by cacodylic acid might play a role in the pathogenesis of its hepatotoxicity.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Chemical Composition of the Pepper Fruit (Dennettia tripetala) Seed Flour
Authors: Oyetayo Folake L. , Ogundare Opeyemi L.
Published: June 2021

The seed flour of the Pepper fruit (Dennettia tripetala) plant was analyzed for the amino and fatty acids, minerals and antinutrient compositions using standard methods. The total amino acid concentration of the seed flour was 83.5g/100g protein of which the total Essential amino acids concentration was 34.3g/100gprotein. Arginine (9.94g/100gprotein) was the most concentrated Essential Amino acid while Glutamic acid(16.04g/100gprotein) and Aspartic acid (15.61g/100g protein) were the most concentrated Non‐Essential Amino acids in the seeds. The lysine/arginine ratio obtained(0.13) suggests that D.tripetala seed possesses a low arteriogenic potential while the calculated P‐PER was 1.91. Linoleic acid was found to be the most abundant fatty acid in the seed oil (38.95%) while the total unsaturated fatty acid concentration (91.29%) was higher than the total saturated fatty acid concentration (8.68%). The oleic/linoleic acid ratio of the seed oil of 1.12 implies the high stability of the seed oil. Potassium (9.31±0.0.05) was the most dominant mineral element (mg/g) while manganese (0.02±0.30) was the least concentrated. Antinutrients were distributed in the seed flour in varying concentrations; phytate (1.20±0.16 mg/g), alkaloids (0.33±0.01%) and flavonoids (2.10±0.01%). The foregoing shows D. tripetala seed as a potential source of useful nutrients which have beneficial implications for human nutrition and health.

Support or Funding Information

nil

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exploration of a pho13 knockout growth phenotype in non‐engineered strains of Saccharomyces cerevisiae
Authors: Ulerio‐Nunez Franly , Gjestvang Mark , O'Handley Suzanne F. , Gehret Austin U.
Published: June 2021

Pho13p from Saccharomyces cerevisiae (budding yeast) is a member of the Haloacid Dehalogenase (HAD) superfamily. Accumulating evidence indicates a loss of Pho13p function imparts an enhanced ability of engineered strains to grow on D‐xylose. Recent evidence has also indicated that Pho13p exhibits specificity towards the substrate 2‐phosphoglycolate but a growth phenotype in non‐engineered pho13 deletion strains has not been apparent. There is evidence that 2‐phosphoglycolate is generated by yeast pyruvate kinase (PYK1) as a secondary metabolic reaction and that fructose‐1,6‐bisphosphate accelerates this activity. The accumulation of 2‐phosphoglycolate under these conditions could impart a growth phenotype in pho13 deletion cells as 2‐phosphoglycolate inhibits triose phosphate isomerase (TPI1). Non‐engineered strains of yeast were subjected to the metabolic inhibitors pyrazole and 2‐thenoyltrifluoroacetone (TTA) that have been shown to increase fructose‐1,6‐bisphosphate in cells. Mutant pho13 deletion strains did not exhibit any hypersensitivity to these inhibitors under rich growth conditions. Currently, pho13 deletion strains are being examined using minimal growth media in combination with pyrazole and TTA. The yeast pyruvate kinase open reading frame is also being PCR‐amplified from genomic DNA and cloned under the control of the GAL1 promoter to assess whether overexpression could enhance a potential inhibitor effect.

Support or Funding Information

This work was supported by an internal NTID FEAD grant (A.U.G.)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification and Regulation of NRNP1, a Novel Nutritionally Regulated Gene Involved in Cell Death
Authors: Cairl Nicholas Stephen , Kalman‐Maltese Vivian , Smas Cynthia M.
Published: June 2021

Obesity and Type II diabetes are co‐morbidities that are increasing at an alarming rate in the United States. A thorough understanding of the genes expressed in white adipose tissue (WAT) and the role of these genes in energy metabolism has high relevance to development of new approaches to treat diseases of dysregulated adipose tissue function. Here, we report a novel adipocyte‐enriched gene NRNP1 (Nutritionally Regulated Nuclear Protein 1) which we discovered via in silico data‐mining of whole transcriptome profiling studies of in vitro adipogenic conversion of murine 3T3‐L1 preadipocytes to adipocytes. Ectopic expression studies of NRNP1‐eGFP fusion protein in COS7 cells revealed strong localization of NRNP1 to the cell nucleus, as well as punctate cytoplasmic localization. Western blot analysis of cell lysates from 293T cells transfected with Flag‐tagged NRNP1 indicated a mass of ~22 kDa. Interestingly, such studies revealed that ectopic expression of NRNP1 in 293T cells resulted in cell death with an ~80% reduction in cell number at 24 hrs post‐transfection. The underlying mechanism for this was investigated utilizing western blot studies for apoptotic marker proteins and with DNA fragmentation analysis. In adipogenesis NRNP1 is under strong developmental control with NRNP1 transcript induced > 500‐fold during conversion of 3T3‐L1 preadipocytes to adipocytes. A similar high degree of differentiation‐dependent induction of NRNP1 transcript was observed in an in vitro model of brown adipogenesis and in adipogenesis of primary murine preadipocytes. Strong nutritional regulation of NRNP1 was noted in vivo as NRNP1 transcript was significantly increased in subcutaneous WAT (12‐fold) and in brown adipose tissue (BAT) (3‐fold) of mice that were fasted and then refed vs. levels in fasting mice. The observation of a 12‐fold increase in NRNP1 transcript level in fatty liver of ob/ob mice vs. C57BL/6J mice suggests increased NRNP1 levels may be linked to dysregulated lipid metabolism. Assessment of a broad panel of mouse tissues by qPCR indicated selective enrichment of NRNP1 in WAT depots and BAT. Together, these data support a role for NRNP1 in normal and abnormal lipid metabolism and indicate that mis‐expression of NRNP1 in non‐adipose cells can promote cell death.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
PI(3,5)P2 Regulates Vacuole Cation Transport to Mediate Cellular Osmoregulation
Authors: Wilson Zachary , Odorizzi Greg
Published: June 2021

Lysosomes are dynamic organelles with critical roles in cellular physiology. The lysosomal signaling lipid phosphatidylinositol 3,5‐bisphosphate (PI(3,5)P2) is a key regulator that has been implicated to control lysosome ion homeostasis, but the scope of ion transporters targeted by PI(3,5)P2 and the purpose of this regulation is not well understood. We identified loss‐of‐function mutations in the vacuolar H+‐ATPase (V‐ATPase) and in Vnx1, a vacuolar monovalent cation/proton antiporter that suppress the growth defects and osmotic swelling of vacuoles (lysosomes) in Saccharomyces cerevisiae lacking PI(3,5)P2 synthesis. The loss of PI(3,5)P2 synthesis in yeast causes a robust accumulation of multiple cations, most notably a ~3‐fold increase in cellular potassium, a primary osmolyte that regulates cellular/organellar water content. The accumulation of potassium and other cations in yeast lacking PI(3,5)P2 is relieved by Vnx1 or V‐ATPase inactivation, or by increased activation of a vacuolar cation export channel, Yvc1. Furthermore, inactivating the V‐ATPase or Vnx1 rescued several vacuole functions that are defective in PI(3,5)P2‐deficient cells. Thus, our data demonstrates that a critical function of PI(3,5)P2 signaling is to orchestrate vacuole/lysosome ion transport to maintain cellular osmoregulation and lysosome homeostasis.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Fic‐mediated deAMPylation is not dependent on homo‐dimerization and rescues toxic AMPylation in flies
Authors: Casey Amanda K. , Moehlman Andrew T. , Zhang Junmei K. , Servage Kelly A. , Krämer Helmut K. , Orth Kim
Published: June 2021

Protein chaperones play a critical role in proteostasis. The activity of the major endoplasmic reticulum chaperone BiP (GRP78) is regulated by Fic‐mediated AMPylation during resting states. By contrast, during times of stress, BiP is deAMPylated. Here, we show that excessive AMPylation by a constitutively‐active FicE247G mutant is lethal in Drosophila. This lethality is cell autonomous as directed expression of the mutant FicE247G to the fly eye does not kill the fly but rather results in a rough and reduced eye. Lethality and eye phenotypes are rescued by the deAMPylation activity of wild‐type Fic. Consistent with Fic acting as a deAMPylation enzyme, its activity was both time and concentration dependent. Furthermore, Fic deAMPylation activity was sufficient to suppress the AMPylation activity mediated by the constitutively‐active FicE247G mutant in Drosophila S2 lysates. Further, we show that the dual enzymatic activity of Fic is, in part, regulated by Fic dimerization as loss of this dimerization increases AMPylation and reduces deAMPylation of BiP. We believe this mechanism leads to a molecular rheostat in the cell for induction of the UPR.

Support or Funding Information

This work was funded by National Institutes of Health (NIH) grant R01 GM115188, the Welch Foundation grant I‐1561, and Once Upon a Time…Foundation to K.O., NIH grants (EY010199 and EY021922) to H.K. and NSF Graduate Research Fellowship (1000176311) to A.T.M. K.O. is a Burroughs Welcome Investigator in Pathogenesis of Infectious Disease, a Beckman Young Investigator, and a W. W. Caruth, Jr., Biomedical Scholar and has an Earl A. Forsythe Chair in Biomedical Science.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The loop diuretics bumetanide and ethacrynic acid inhibit voltage‐gated calcium and sodium channels in cultured cortical neurons
Authors: Katnik Christopher , Cuevas Javier
Published: June 2021

One of the consequences of ischemic stroke is disruption of intracellular ionic homeostasis. Intracellular overload of both Na+ and Ca2+ have been linked to neuronal death in this pathophysiological state. The etiology of ionic imbalances resulting from stroke‐induced ischemia and acidosis includes the dysregulation of a number of plasma membrane transport proteins, such as increased activity of the sodium‐potassium‐chloride cotransporter 1 (NKCC1). Experiments using the NKCC1 inhibitors, bumetanide (BMN) and ethacrynic acid (EA), were carried out to determine if inhibition of this cotransporter affects the Na+ and Ca2+ overload observed following in vitro ischemia‐acidosis. Fluorometric Ca2+ and Na+ measurements were performed on cultured cortical neurons from embryonic (E18) rats. Both BMN and EA depressed ischemia‐acidosis induced [Ca2+]i overload, but did not appreciably alter [Na+]i increases observed under these conditions. Patch‐clamp electrophysiology was used to measure whole‐cell membrane currents to identify target(s) of BMN and EA, other than the electroneutral NKCC‐1, responsible for the effects of these drugs on [Ca2+]i. Voltage‐gated Ca2+ channels (VGCC) were inhibited by both BMN and EA with half‐maximal inhibitory concentration (IC50) values of 4 μM and 36 μM respectively. However, maximal concentrations of BMN failed to block ~40% of VGCC mediated currents, while EA completely abolished these responses. Similarly, voltage‐gated Na+ channels (VGSC) were blocked by BMN and EA with IC50 values of 13 μM and 30 μM, respectively. In contrast to the VGCC responses, both loop diuretics inhibited VGSC currents by ~80%. Experiments using the VGSC inhibitor, tetrodotoxin, showed that BMN was preferentially inhibiting TTX‐sensitive currents in these cells. Neither BMN nor EA affected currents mediated by acid‐sensing ion channels or ionotropic glutamatergic receptors. Taken together, our data suggest that these two loop diuretics are likely to decrease injury following ischemic stroke by effectively mitigating [Ca2+]i overload in neurons via the inhibition of both voltage‐gated Ca2+ and Na+ channels. Moreover, the depression of ischemia‐acidosis evoked [Ca2+]i overload by these compounds does not appear to be dependent on inhibition of NKCC1. Given that the inhibitory concentrations observed here for VGCC and VGSC are similar to those reported for block of NKCC1 and NKCC2 by these compounds, loop diuretics are likely to be inhibiting these channels at therapeutic doses used in humans. Therefore, blockade of these channels by loop diuretics may also contribute to other clinical effects of the compounds observed in animal studies and in humans.

Support or Funding Information

American Heart Association Grant‐In‐Aid, 17GRNT33661273 (JC)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of soluble epoxide hydrolase attenuates the kidney injury caused by ischemia/reperfusion in a murine model of acute kidney injury involved in GSK‐3β phosphorylation
Authors: Liu Jun‐Yan , Deng Bing‐Qing , Luo Ying , Kang Xin , Li Chang‐Bin , Lee Kin Sing Stephen , Hammock Bruce D.
Published: June 2021

Acute kidney injury (AKI) causes severe morbidity and mortality for which new therapeutic strategies are needed. Inhibition of soluble epoxide hydrolase (sEH) has been reported previously to attenuate the kidney injury in cisplatin‐caused AKI. However, the effect of sEH inhibition on ischemia/reperfusion (I/R)‐caused‐AKI remains unknown. In addition, the mechanism underlying the benefit of sEH inhibition to AKI has not been well understood. Here we report that epoxyeicosatrienoic acids (EETs) are the major mediators responsible for the beneficial effect of sEH inhibition in I/R‐caused AKI. Inhibition of sEH resulted in a significant increase in plasma level of 14(15)‐EET. Administration of both 14(15)‐EET alone and TPPU (a potent sEH inhibitor) significantly prolonged the survival of AKI mice. Co‐administration of 14(15)‐EET with TPPU enhanced such effect in survival. Administration of 14(15)‐EET with or without TPPU to AKI mice significantly attenuated the kidney injury, which was supported by the histological analysis of renal tissue, plasma levels of creatinine and urea nitrogen, and renal levels of NGAL. Furthermore, 14(15)‐EET significantly reversed the I/R‐caused reduction in glycogen synthase kinase 3β (GSK3β) phosphorylation in murine kidney, dose‐dependently inhibited the hypoxia/reoxygenation (H/R)‐caused apoptosis of murine renal tubular epithelial cells (mRTECs), and reversed the H/R‐caused reduction in GSK3β phosphorylation in mRTECs. Therefore, inhibition of sEH attenuates kidney injury via, at least partially, 14(15)‐EET modulating the apoptosis of RTECs and GSK3β phosphorylation in mRTECs. This study provides AKI patients with promising therapeutic strategies.

Support or Funding Information

This study was supported in part by NSFC grants 81470588 and 81100090, as well as NIEHS Grant (R01 ES02710), NIEHS Superfund Grant (P42 ES04699), NIH/NHLBI grant (R01 HL59699‐06A1), and a Translational Technology Grant from the UC Davis Medical Center. K.S.S.L. is supported by NIEHS Grant R00 ES024806.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
MUSCADINE GRAPE SKIN EXTRACT INHIBITS ANDROGEN‐INDEPENDENT PROSTATE CANCER CELL GROWTH, INDUCING CELL‐CYCLE ARREST, AND DECREASING MIGRATION BY TARGETING HEAT SHOCK PROTEIN 40
Authors: Brown Collis
Published: June 2021

Previously we demonstrated that muscadine grape skin extract (MSKE), a natural product, significantly inhibited prostate cancer cell growth by inducing apoptosis through the targeting of phosphatidylinositol 3‐kinase‐Akt and mitogen‐activated protein kinase survival pathways in androgen‐responsive transformed human prostate cancer epithelial cell lines. However, the preventive effect of MSKE on more aggressive androgen‐independent prostate cancer remains unknown. This study examined the effects of MSKE treatment using complementary PC‐3 prostate cancer cell culture and xenograft models. MSKE significantly inhibited PC‐3 human prostate cancer cell tumor growth in vitro and in vivo. The growth‐inhibitory effect of MSKE appeared to be through the induction of cell‐cycle arrest. This induction was accompanied by a reduction in the protein expression of Hsp40 protein and cell‐cycle regulation proteins, cyclin D1 and NF‐kBp65. In addition, MSKE induced p21 expression independent of wild‐type p53 induced protein expression. Moreover, we demonstrated that MSKE significantly inhibited cell migration in PC‐3 prostate cancer cells which was complimented by siRNA‐HSP40 knock‐down that also inhibited cell migration and growth. Overall, these results demonstrate that MSKE inhibits prostate tumor growth and migration, and induces cell‐cycle arrest by targeting Hsp40 and proteins involved in cell‐cycle regulation and proliferation. This suggests that MSKE may also be explored as a novel therapeutic in castration resistant prostate cancer.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The α7‐nAChRs/heme oxygenase/carbon monoxide pathway arbitrates nicotine counteraction of the inflammatory and renal vasoconstrictor hyporeactivity in endotoxic rats
Authors: Wedn Abdalla M. , El‐Gowilly Sahar M. , El‐Mas Mahmoud M.
Published: June 2021

The nicotinic/cholinergic antiinflammatory pathway plays integral roles in physiological and pathological regulation of inflammation. In this study, we tested the hypotheses that (i) nicotine guards against hemodynamic and renal vasoconstrictor dysfunction induced by endotoxemia in rats, and (ii) the α7‐nAChRs/heme oxygnase (HO) pathway constitutes a go‐between for the LPS‐nicotine interaction. Hemodynamic or renovascular functions were assessed 6 hr after i.p. administration of lipopolysaccharide (LPS, 5 mg/kg) with or without nicotine. LPS caused significant falls in systolic blood pressure (SBP) and reductions in cumulative renal vasoconstrictions induced by bolus injections of phenylephrine (PE, 0.002 – 4.42 nmol) or vasopressin (VP, 0.00092 – 0.27 nmol) into isolated perfused kidneys. LPS hypotension was demonstrated in both male and female rats, whereas the attenuation of renal vasoconstrictions was seen in male rats only. The reduced PE or VP responsiveness was reversed in preparations pretreated with nicotine (0.5, 1 and 2 mg/kg) in a dose‐dependent manner. This favorable action of nicotine disappeared after blockade of α7‐nAChRs (methyllycaconitine citrate, 2 mg/kg) or inhibition of HO‐1 (zinc protoporphyrin, 10 mg/kg). Moreover, the LPS attenuation of renal vasoconstrictions was improved upon concurrent treatment with pentoxfylline (TNFα inhibitor, 3 mg/kg), hemin (HO‐1 inducer, 10 mg/kg), tricarbonyldichlororuthenium (II) dimer (carbon monoxide‐releasing molecule, 10 mg/kg), in contrast to no effect for bilirubin (5 mg/kg). These data establish the first evidence that implicate the α7‐nAChRs/HO‐1/CO in the nicotine counteraction of renal vasoconstrictor hyporeactivity in endotoxemia.

Support or Funding Information

Supported by the Science and Technology Development Fund, Egypt (STDF Grant No. 14895)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Aortic Function of Female UC Davis Type 2 Diabetes Mellitus (UCD‐T2DM) Rats
Authors: Akther Farjana , Razan Md Rahatullah , Tang Tia , Graham James L. , Stanhope Kimber L. , Havel Peter J. , Rahimian Roshanak
Published: June 2021

UC Davis Type 2 Diabetes Mellitus (UCD‐T2DM) rat is a novel validated model of type 2 diabetes mellitus. UCD‐T2DM is characterized by polygenic, adult‐onset obesity and spontaneous β‐cell failure and, as a result, more closely models the pathophysiology of type 2 diabetes in humans than other rodent models. The objectives of this study were to determine the aortic function of female UCD‐T2DM rats and to investigate the underlying mechanism(s) of altered vascular function (if any) in this model. Endothelium‐dependent vasodilation (EDV) to acetylcholine (ACh, 10−8 to 10−5M) was measured in intact aortic rings pre‐contracted with phenylephrine (PE, 2μM). Endothelium‐independent vasodilation to sodium nitroprusside (SNP, 10−9 to 10−5 M) was assessed in endothelium‐denuded rings pre‐contracted with PE (2μM). Furthermore, constrictor response curves to PE (10−8 to 10−5 M) were generated before and after incubation with L‐NAME (200μM), a nitric oxide synthase (NOS) inhibitor. Expression of molecules associated with vascular reactivity and insulin signaling were also evaluated using Western blotting in aortic tissues.

We demonstrated that the maximal relaxation to ACh was significantly reduced in aortic rings from female UCD‐T2DM rats compared to controls. The smooth muscle sensitivity to NO as measured by SNP‐induced relaxation was also reduced in this group. The responsiveness to PE was significantly enhanced in aorta of UCD‐T2DM group. Accordingly, the basal nitric oxide (NO), as indicated by the potentiation of the response to PE after L‐NAME, was reduced in diabetes group. Moreover, expression of eNOS, peNOS, pAkt, IRS1 and IRS2 were significantly reduced in the aorta of this disease model.

These data, for the first time, show that the vascular function is altered in aortic rings of female UCD‐T2DM rats. Specifically, our data suggests that the decreased sensitivity of vascular smooth muscle to NO along with the enhanced contractile responsiveness to PE may in part contribute to the attenuated ACh responses in aorta of UCD‐T2DM female rats.

Support or Funding Information

Supported by NIH/NIHLBI

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 anesthetic effects of isoflurane used alone or combined with xylazine on induced cerebral ischemia
Authors: Kim Joo‐Heon , Won Jinyoung , Hong Yonggeun
Published: June 2021

Intraluminal suture method of middle cerebral artery occlusion (MCAo) has been extensively used in ischemic stroke study. Although the MCAo model has several advantages in comparing different stroke model, it has time‐consuming demands and requires high level of surgical skill. Low mortality and reproducible infarct size in MCAo model are affected by surgical technique, environmental variation and prolonged duration of surgery. The purpose of this study was to determine the effect of anesthetics during prolonged experimental surgery on reproducible stroke model. Animals were subjected to MCAo using intraluminal suture method and provide with isoflurane anesthesia or combined xylazine sedation (20 mg/kg). Animals were randomly divided into three groups: no anesthesia groups (control), inhaled anesthetics groups (isoflurane), inhaled anesthetics combined sedative group (isoflurane + xylazine). Inhaled anesthetics group was provided isoflurane during MCAo surgery (20 min), post‐occlusion period (60 min) and reperfusion (5–10 min), thus total duration of inhaled anesthesia was about 90 min. Inhaled anesthetics combined sedative groups was provided isoflurane during MCAo surgery (20 min) and reperfusion (5–10) min. Xylazine (20 mg/kg) was administered 5 min prior to MCAo and 30 min after MCAo by intramuscular injection. That total duration of inhaled anesthesia was about 30 min. Inhaled anesthetics group that anesthetized during all the post‐occlusion period demonstrated a poor prognosis and high mortality rate compared with control and inhaled anesthetics combined xylazine group. Although the stable respiration signal was shown without abnormal thorax movement, brain infarct size and neurologic deficits revealed no difference between groups. Our results indicated that a modified method of inhalation anesthesia combined xylazine sedation could improve the reproducibility of stroke model and reduce the risk of mortality.

Support or Funding Information

Funding: NRF‐2013R1A2A2A01067169, NRF‐2017R1A2A2A01067169, KRIBB‐KGM4611714

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inducible Adipose Tissue VEGF‐D Drives Lymphatic Expansion and Improves Systemic Insulin Sensitivity in Obesity
Authors: Chakraborty Adri , Lammoglia Gabriela M. , Rutkowski Joseph M.
Published: June 2021

The metabolic syndrome etiology finds its roots in inflammatory adipose tissue expansion. Rapid adipose expansion, adipocyte insulin resistance, and tissue hypoxia result in dysregulated lipolysis and immune cell infiltration. Increased circulating cytokines and elevated lipid levels lead to ectopic lipid deposition and systemic insulin resistance. Lymphatic vessels regulate inflammation by clearing fluid, cytokines, antigens, and immune cells from the site of inflammation. We hypothesized that increased lymphatic vessel density in adipose tissue may ameliorate local inflammation and thus protect against the metabolic system in obesity. We generated transgenic mice capable of inducible, tissue‐specific expression of murine VEGF‐D under a tightly‐controlled tetracycline response element (TRE) promoter to stimulate lymphangiogenesis. Crossed with adipocyte‐specific adiponectin‐rtTA mice (Adipo‐VD), VEGF‐D overexpression by adipocytes induced de novo lymphangiogenesis in murine white and brown adipose tissues. Adipo‐VD mice gained equivalent weight on high fat diet feeding to their littermates, but demonstrated improved insulin sensitivity and elevated adipose metabolism. b‐adrenergic stimulation of Adipo‐VD mice white adipose lymphatic network promoted enhanced lipolysis and postulated a new route for increased glycerol flux into the circulation: lymphatic vessels. Beyond the increased lymphatic vasculature, adipose tissue architecture and adipocyte size were the same. While chemokine levels in the tissues were unchanged, we quantified fewer macrophage crown‐like structures and reduced macrophages and Tcells in the tissue. Quantitation of infused CD45.1+ leukocytes population in the CD45.2+ Adipo‐VD subcutaneous adipose tissue also indicated increased trafficking to the inguinal lymph node. Increased adipose tissue lymphatic density thus increases insulin sensitivity and enhances immune cell flux from expanded adipose tissue making lymphatics a novel target in the metabolic syndrome.

Support or Funding Information

Lipedema Foundation

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The prevention of cardiometabolic risk factors is associated with the improvement in aerobic capacity
Authors: Vecchiatto Bruno , Martucci Luiz Felipe , Américo Anna Laura Viacava , Muller Cynthia Rodrigues , Evangelista Fabiana Sant'anna
Published: June 2021

Physical inactivity reduces aerobic capacity and is directly associated with an increase in cardiometabolic risk factors. Here we investigated whether there is an association between increased aerobic capacity and the prevention of obesity and insulin resistance through aerobic exercise training (AET). Adult male C57BL6/J mice were assigned into chow‐fed controls (C, n=10), cafeteria diet (CAF, n=8), chow‐fed trained (T, n=9), and cafeteria diet plus trained (CAFT, n=8) groups. AET was performed simultaneously with diet and consisted of 8‐wk running session of 60 min at 60% of maximal speed, 5 days/wk. Experimental procedures were approved by Ethics Committee from School of Arts, Science and Humanities, University of Sao Paulo, Brazil (#001/2016). Both trained groups showed lower body weight at the end of protocol compared to C and CAF groups. However, only CAFT group had reduced body weight gain compared to the CAF group. Cafeteria diet induced glucose intolerance and insulin resistance in the CAF group, which was counteracted by AET in CAFT group. Indirect calorimetry measurement at rest revealed no difference in the variables oxygen uptake (VO2), carbon dioxide expiration (VCO2), respiratory exchange ratio (RER), energy expenditure (EE), carbohydrate and fat oxidation among groups. During maximal exercise tests, T and CAFT groups showed an increase in VO2máx (13% and 9.2%, p=0.07) and VO2peak (11.3% and 8.6%, p=0.07) compared to C and CAF, respectively. However no differences among groups were found for RER and rate of substrate oxidation. AET increased the time to exhaustion and maximal velocity in the T compared to C and CAF groups, and in the CAFT compared to CAF. The exercise intensity corresponding to the VO2máx (iVO2) was higher in the T group (2.30 ± 0.42 km⁄h) compared to C (1.86 ± 0.41 km/h) and CAF (1.73 ± 0.17 km/h) groups, and in the CAFT (2.23 ± 0.11 km/h) compared to CAF. Both trained groups showed late anaerobic threshold, improved the relative cost of running, and higher activity of citrate synthase and β‐HAD enzymes in the soleus muscle compared to C and CAF groups. In conclusion, these results revealed that the prevention of obesity and insulin resistance is associated with the improvement in aerobic capacity induced by AET.

Support or Funding Information

Financial Support: FAPESP # 2015/04948‐4; 2016/20659‐5.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Relationship between Injury and Spirituality in Pilgrims on the Camino de Santiago
Authors: Rennyson Katelyn Elizabeth , Harris M. Brennan
Published: June 2021

The purpose of this research was to determine whether there is a relationship between spirituality and the likelihood of injury and the impact of that injury on pilgrims walking along the Camino de Santiago; an ancient, 791 km pilgrimage route along the north of Spain from the French Pyrenees to Santiago de Compostela in Galicia, Spain.

400 participants aged 18–80 were recruited along the Camino and through social media prior to and during the pilgrimage while on their journey towards Santiago. Participants completed a ten‐minute survey with questions pertaining to general demographics, injury and illness experiences on the walk, physical well‐being, and fitness. Spirituality was assessed using the FACIT‐Sp questionnaire. Data was analyzed using Excel and Qualtrics software. Research was approved by the Protection of Human Subjects Committee at William & Mary and complies with the principles and protections endorsed by Experimental Biology regarding the use of human subjects and informed consent.

Results show that participants who scored a higher spirituality score were less likely to report that any injuries they experienced on the pilgrimage impacted their experience along the trail. The data also demonstrated that participants who were more spiritual were less likely to experience an injury while walking the pilgrimage.

These data suggest that pilgrims who walk the Camino de Santiago for spiritual or religious reasons may experience less injuries overall and will feel less impacted by those injuries.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Dopamine D2 receptor decreases the toxic effects of aristolochic acid in human renal proximal tubule cells.
Authors: Kumar Megha , Ayvazian Nicholas , Gildea John , Felder Robin A. , Jose Pedro A. , Armando Ines
Published: June 2021

Previous work from our laboratory indicates that the renal dopamine D2 receptor (D2R) has a direct role in regulating renal inflammation and injury and blood pressure. In mice, renal‐selective D2R silencing increased the expression of pro‐inflammatory and pro‐fibrotic factors including TNFα, chemokine CCL‐2, TGFβ, fibronectin 1 (Fn1), and collagen 1α1 (Col1α1). Moreover, the damaging effects of 45‐minute bilateral ischemia/reperfusion on renal expression of profibrotic factors, renal function, and blood pressure in mice were ameliorated by a renal‐selective increase in D2R expression by the retrograde ureteral infusion of a viral vector carrying DRD2. In human renal proximal tubule cells (hRPTCs), D2R silencing increased the expression of TGFβ, Fn1 and Col1α1. We tested the hypothesis that in hRPTCs, D2R agonism decreases the deleterious effects of aristolochic acid, a potent nephrotoxic. hRPTCs were treated with aristolochic acid (5μg/ml, 24 hrs) in the presence or absence of the D2R agonist quinpirole (0.5 μM). Treatment with aristolochic acid increased the mRNA expression of TGFβ (1.3 ± 0.02; n=3; P<0.05), Fn1 (2.6 ±0.4‐fold, n=3, P<0.05), Col1α1 (1.9±0.2‐fold; n=3; P<0.05), and connective tissue growth factor (CTGF, 7.6 ± 0.3‐fold, n=3, P<0.05). Treatment with quinpirole, alone, decreased significantly the mRNA expression of TGFβ (0.7±0.05‐fold, n=3, P<0.05) and Fn1 (0.8±0.01‐fold, n=3, P<0.05) but did not change the expression of Col1α1, or CTCG. However, quinpirole reduced significantly the aristolochic‐mediated increase in the expression of Fn1 by 43% and Col1α1 by 35% but had no significant effect on that of TGFβ or CTGF. Our results indicate that several pathways are involved in the nephrotoxic effects of aristolochic acid and that D2R agonism is able to negatively regulate some but not all of those pathways.

Support or Funding Information

This work was funded by grants from the NIH: R01DK090918, P01HL074940, P01HL068686, R01HL092196, R37HL023081, and R01DK039308.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of Buffer Composition and Substrates on Calcium‐stimulated Mitochondrial Respiration and Free Radical Emission
Authors: Duong Quynh , Zhong Katie , Zhang Yizhu , Bazil Jason
Published: June 2021

Mitochondrial ATP synthesis is tightly coupled to oxygen consumption in a highly regulated process; however, unavoidable byproducts in the form of reactive oxygen species (ROS) are formed. An important regulator of this process is calcium, but the precise relationship between calcium and free radical production is obscure. Therefore, we measured ROS production under a variety of experimental conditions that mimic the pathological environmental setting. The Oroborous oxygraph system was used to simultaneously measure ROS emission and the oxygen consumption rate from isolated cardiac guinea pig mitochondria energized with different substrates and challenged with different calcium concentrations. The Amplex Red assay was used to quantify ROS emission rates. The substrates used utilized a mixture of the carbohydrate and fatty acid dependent metabolic pathways. Two respiration buffers, one lactobionate‐based and the other potassium chloride‐based, were used to quantify the differences in respiration and ROS emission rates. Relative to the potassium chloride‐based respiration buffer, the lactobionate‐based buffer resulted in higher ADP‐stimulated respiration rates and higher ROS emission rates for each substrate combination. On average, the presence of calcium depressed ROS emission rates during the leak respiration, sodium/calcium cycling, and ADP‐stimulated states. The lower respiration rates in the potassium chloride‐based buffer is likely due to the presence of inhibitory levels of chloride. The high chloride levels also appear to depress ROS emission. Overall, these data refute the hypothesis that calcium stimulates ROS emission from isolated mitochondria. Studies are ongoing to further characterize ROS emission from cardiac guinea pig mitochondria and elucidate the relationship between calcium and ROS emission.

Support or Funding Information

R00‐HL121160

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.