bluuebunny/crossref_metadata_2025 huggingface dataset:
Chlorogenic Acid Enriched in Coffee Pulp Extract Attenuates Hepatic Steatosis by Modulating Gene‐regulated Lipid Metabolism and Lipid Transporters
Authors: Ontawong Atcharaporn , Pasachan Tipthida , Soodvilai Sunhapas , Duangjai Acharaporn , Pongchaidecha Anchalee , Amornlerdpison Doungporn , Srimareong Chutima
Published: June 2021

Background & Aims

Coffee pulp (CP) is the first by‐product of coffee processing. Major constituents in CP include chlorogenic acid (CGA), epicatechin (EC), caffeine exhibited antioxidant, lipid lowering action, and several biological activities. However, it remains unknown whether CP rich CGA improved hepatic steatosis. This study aimed to investigate the effect of CP aqueous extract (CPE) on hepatic steatosis, and identified the mechanisms involved in hepatocellular carcinoma (HepG2) cells.

Methods

HepG2 cells at the cell density of 5×105 cells/well were pre‐incubated with CPE (10–200 μg/ml), CGA, and caffeine for 24 hr. Intracellular lipid accumulation was quantified using Nile red staining followed by flow cytometry. Cell viability of HepG2 cells was determined using MTT assay. The mRNA expression of genes involved in lipid metabolism and lipid transporters were also determined using quantitative real‐time PCR.

Results

CPE at the dose of 50–200 μg/ml, CGA, and caffeine significantly decreased lipid accumulation in HepG2 cells. Correspondingly, CPE decreased lipogenesis‐associated genes: PPARγ, and up‐regulated lipolysis‐related genes: PPARα. In addition, CPE reduced the lipid uptake transporter, FAT/CD36, while increased the lipid efflux transporters, ABCG5/8.

Conclusion

CPE has hepatic lipid‐lowering effects by modulating genes related lipolysis, lipogenesis, and lipid transporters. These findings indicate that anti‐hepatic steatosis by CPE is attributed to hepatic lipid‐lowering action for reversing such pathology. Thus, CPE could potentially be developed into nutraceutical product for prevention of non‐alcoholic fatty liver disease.

Support or Funding Information

This work was supported by the Faculty of Medicine Endowment Fund (066/2560 to CS), Chiang Mai University, the Research and Researcher for Industry (RRI) by Thailand Research Fund (TRF) (PHDI0010 to CS), and the NSTDA Chair Professor grant (the Fourth Grant) of the Crown Property Bureau Foundation and the National Science and Technology Development Agency to Professor Dr. Vatcharin Rukachaisirikul.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Oligomerization of ferroportin and the mechanism of autosomal dominance in ferroportin disease
Authors: Bonamer John P. , Ruwe T Alex , Qiao Bo , Vieth Kyle R. , Ganz Tomas , Nemeth Elizabeta , Mackenzie Bryan
Published: June 2021

Ferroportin (Fpn)—the only known cellular iron‐export protein—plays a critical role in iron homeostasis by mediating iron export from macrophages and enterocytes. Rare, loss‐of‐function mutations in human Fpn (e.g. V160del) cause classical ferroportin disease, characterized by macrophage iron loading and iron‐restricted anemia. Ferroportin disease displays an autosomal–dominant inheritance pattern. Because all of the disease‐associated mutations are missense, the dominant–negative effect is most readily explained as resulting from oligomerization of Fpn protein. As a first step in testing this hypothesis, we have examined the functional activity of wildtype (wt) and mutant human Fpn co‐expressed in RNA‐injected Xenopus oocytes. We measured 55Fe efflux from control oocytes, or oocytes expressing wtFpn or V160del GFP proteins with or without co‐expression of a wtFpn–mCherry fusion protein. The RNA dose eliciting half‐maximal iron‐export activity was 12.1 ± (SEM) 1.6 ng for wtFpn–GFP (P = 0.005) and 11.7 ± 2.2 ng for wtFpn–mCherry (P = 0.013), so we injected 12 ng RNA of each construct in subsequent co‐expression experiments. 55Fe efflux activity resulting from co‐expression of wtFpn–GFP and wtFpn–mCherry constructs was additive (Fig 1). Whereas expression of V160del–GFP alone did not stimulate 55Fe efflux, co‐expression of the mutant with wtFpn–mCherry stimulated 55Fe efflux to roughly the same degree as either of the wildtype constructs alone. Our results therefore provide no evidence of a dominant–negative effect of V160del. Our next step will be to test co‐expression of wtFpn with other loss‐of‐function mutants. We will examine the oligomeric structure of wildtype and mutant Fpn by using freeze‐fracture electron microscopy to examine the interior (p) face of the inner lamella of the plasma membrane, as previously described for several membrane‐transport proteins [Eskandari S et al (1998) Proc Natl Acad Sci USA 95, 11235–11240]. We anticipate that our study will further elucidate the structure–function of Fpn and help to explain the autosomal dominance of ferroportin disease.

Support or Funding Information

NIH–NIDDK grant R01 DK107309

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Regulation of Intestinal Phosphate Transport in Humans and in a Rat Model of Chronic Kidney Disease
Authors: Myakala Komuraiah , Wang Xiaoxin , Levi Moshe
Published: June 2021

In chronic kidney disease (CKD) hyperphosphatemia is a common occurrence and plays important roles in cardiovascular and bone disease. The mechanisms however still remain unknown and the role of intestinal phosphate (Pi) transport is subject of ongoing debate. We have determined that Na/Pi cotransport activity and NaPi‐2b protein is expressed in the duodenum, jejunum, and ileum of human intestinal samples. We further studied regulation of intestinal phosphate transport in a model of 5/6 nephrectomy (Nx) induced CKD in the rat fed a relatively high Pi diet (1.5% Pi, 0.6% Ca). Male rats with 5/6 Nx had a marked increase in serum BUN (37.7 ± 2.0 vs. 150.5 ± 29.8 mg/dl in sham control, p<0.02), serum creatinine (0.5 ± 0.06 vs. 1.65 ± 0.23 mg/dl in sham control, p<0.008), and serum Pi (7.48 ± 0.85 vs. 18.19 ± 1.84 mg/dl in sham control, p<0.001). We isolated apical brush border membrane vesicles (BBMV) from the duodenum and the jejunum and studied sodium gradient dependent Pi (Na+/Pi) cotransport as a function of pH. Across every single pH studied, including 5.5, 6.0, 6.5, 7.0, and 7.5 Na+/Pi transport activity was increased in BBM from 5/6 Nx rats. This was associated with a 2‐fold increase in NaPi‐2b protein abundance in the duodenum and a1.4‐fold increase in NaPi‐2b protein abundance in the jejunum. To determine if there are sex‐dependent differences in CKD and intestinal Na+/Pi transport, we also studied in parallel female rats with 5/6 Nx. We found that female rats with 5/6 Nx also had a marked increase in serum Pi (4.75 ± 0.39 vs. 10.7 ± 1.24 mg/dl in sham control, p<0.003). In addition, there were marked increases in intestinal Na+/Pi transport activity paralleled by increases in NaPi‐2b protein abundance.

Our study therefore indicates that in both male and female rats with CKD, increased intestinal Na+/Pi transport activity and increased NaPi‐2b protein expression mediate the hyperphosphatemia

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 local environment of cysteine 621 determines the rapid electrophilic adduction and activation of hTRPA1
Authors: Bahia Parmvir K. , Parks Thomas A. , Stevens Stanley M. , Varma Sameer , Taylor‐Clark Thomas E.
Published: June 2021

Transient Receptor Potential Ankyrin 1 (TRPA1) is a non‐selective cation channel expressed largely on nociceptive sensory nerves. TRPA1 is rapidly activated by electrophiles, such as iodoacetamide, through covalent modification of intracellular cysteine (Cys) residues. We have previously shown that, at physiologically relevant exposures, iodoacetamide rapidly labels four Cys residues at positions 273, 621, 655, 1085. Of these, Cys residue at position 621 (C621) appears to be crucial for binding and activation of the human channel (hTRPA1). Interestingly, when comparing hTRPA1 with the electrophile‐insensitive rattlesnake homolog, while the amino acids lysine K620 and C621 are conserved the proline at P622 is not. We hypothesized that this rapid binding of C621 residue in human TRPA1 is mediated by neighboring amino acids K620 and P622 rendering it more sensitive to electrophilic adduction. We expressed V5‐His tagged hTRPA1 constructs in HEK293 cells including the wild‐type and the point mutants C621A, K620A and P622A. We used pulse‐chase experiments to label TRPA1 protein with iodoacetamide‐alkyne (IA‐alk), then used click chemistry to label with a fluorescent tag. Binding was measured using immunoprecipitated TRPA1 separated by SDS‐polyacrylamide gel electrophoresis and western blotting. TRPA1 channel activation was measured using ratiometric calcium imaging. The K620A mutant had the effect of greatly reducing electrophilic binding and activation of the channel. Modeling of deprotonation energies suggests that K620 contributes to C621 reactivity by reducing its pKa. The K620 mutant therefore likely reduces the probability that C621 will be thiolate and therefore able to rapidly react with electrophiles. The P622A mutant was also largely insensitive to electrophilic adduction and activation. IA‐alk binding was ~10% of that seen with the wild‐type channel. We hypothesize this is because, in the wild‐type hTRPA1, P622 induces a kink in the protein resulting in the positively charged K620 being closer to C621 compared with the nearby negatively charged glutamates, E625 and E681. The work described here highlights the importance of the amino acids K620 and P622 flanking C621. Our data demonstrate how the integrity of the local environment is essential to TRPA1 function as an electrophilic sensor.

Support or Funding Information

This work was supported by the National Heart Lung and Blood Institute in Bethesda, USA(R01‐HL119802‐S1)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Age‐dependent Changes in miRNA Profile in F344 rat and C57BL/6J mice: Role of sodium hydrogen exchanger regulatory factor‐1 (NHERF1)
Authors: Jain Anish , Aubee Joseph I. , Thompson Karl M. , Lee Dexter L. , Khundmiri Syed Jalal
Published: June 2021

Our laboratory demonstrated decreased NHERF1 expression in aging F344. Loss of NHERF1 expression prevented dopamine‐dependent inhibition of NKA in renal PTCs from aged F344. We hypothesized that loss of NHERF1 expression in aged F344 rats is due to changes in miRNA expression. To address this hypothesis we analyzed the miRNA expression profile in 4mo and 22mo old F344 rats and 2 and 18mo old C57BL/6J wild‐type and NHERF1 knock‐out mice. Total RNA from kidney cortex was isolated using Trizol (miRVana, Ambion) and miRNA profile was analyzed using kits and software from Nanostring. The top 100 miRNAs with 25% counts above the negative miRNA counts were considered for analysis. Counts from young animals (n=4, 2M2F) were averaged together and considered as 1. Counts from old animals (n=4, 2M2F) were calculated as fold change. In F344 rats 10 miRNAs increased with age (miR29a, miR29b, miR29c, and miR21), while miR96 and miR328a decreased. In WT‐ mice 51 miRNAs increased with age (miR‐let‐7g, b, and d; and miR22), while 28 miRNAs decreased (miR21 and miR‐let‐7e). In NHERF1‐KO mice 31 miRNAs increased with age (miRNA21, miR29a, miR29b, and miR29c), while 22 miRNAs decreased (miR‐let‐7a, miR‐let‐7b, miR‐let‐7c, and miR24). Our results suggest that miRNAs that regulate inflammation increased with age in both F344 rats and NHERF1‐KO mice. Interestingly, these miRNAs decreased in WT mice. These data may explain the mechanisms of increased renal inflammation in NHERF1 deficient models of hypertension.

Support or Funding Information

7R21AG047474‐03 NIH

R25AG047843‐NIH

16GRNT31030019‐AHA

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Gut Microbiota Modulation on Hepatic Lipid Metabolism
Authors: Pereira Louise Tavares Garcia , Beserra Bruna Teles Soares , Engel Daiane Fátima , de Paula Gabriela Cristina , de Bema Andreza Fabro , Amato Angélica Amorim
Published: June 2021

Non‐alcoholic fatty liver disease is characterized by fat accumulation in the liver and commonly seen in patients with components of the metabolic syndrome. The mechanisms underlying its development are not completely defined, but there has been an increasing body of evidence linking gut microbiota to the development and progression of fat accumulation in the liver. Our objective was to investigate the effect of gut microbiota modulation on hepatic lipid metabolism.

Methods

Six week old C57BL/6 male mice, fed with a control diet (CD, n=18) or high‐fat diet (HFD, n=30). At the age of 7 weeks, they were randomly assigned into 5 groups to receive (i) CD and vehicle, (ii) CD and antibiotics, (iii) HFD and vehicle, (iv) HFD and antibiotics, and (v) and HFD and antibiotics followed by fecal transplantation from CD‐ fed donors. Antibiotics (ciprofloxacin 0.2g l−1 and vancomycin 0.5 g l−1) were administered from the 10th to the 15th week, in drinking water. Fecal transplantation was carried out at the 13th week, following antibiotic withdrawal and maintenance of recipient mice on HFD. Weight was measured weekly and water intake twice a week. An oral glucose tolerance test (GTT) was performed at the 12th weeks. Mice were euthanized by decapitation and liver samples were stored for histological analysis and gene expression by real time quantitative polymerase chain reaction.

Results

HFD‐fed mice showed significantly increased weight gain when compared with CD‐fed mice. Antibiotic treatment did not change weight gain in CD‐fed mice but increased weight gain in HFD‐fed mice. Mice fed a CD and treated with antibiotics showed increased glucose tolerance when compared with control (CD, vehicle), and antibiotic treatment of HFD‐fed mice did not change glucose tolerance. CD‐fed mice treated with antibiotics exhibited increased expression of mRNA levels of PPAR‐α in liver and no change in lipogenesis‐related transcripts (Srepb1 and Acsl1), when compared with CD‐fed mice treated with vehicle. Mice fed a HFD and treated with antibiotics showed no change in PPARa, Srepb1 and Acsl1 levels when compared with control (HFD, vehicle), whereas HFD‐fed mice receiving fecal transplantation from CD‐fed mice showed a trend towards increased PPARa levels in the liver.

Conclusion

Our preliminary data may suggest that gut microbiota modulation with antibiotics may affect fatty acid metabolism in the liver by changing the balance between expression of lipogenesis and fatty acid oxidation‐related genes. Further studies are needed to clarify the extension and mechanisms underlying these effects.

Support or Funding Information

This study was supported by the Fundação de Amparo à Pesquisa do Distrito Federal (FAP‐DF).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
CART‐GPR160 Regulation of Food Intake
Authors: Haddock Christopher J. , Yosten Gina L.C. , Kolar Grant R. , Samson Willis K.
Published: June 2021

Cocaine and amphetamine regulated transcript (CART) mRNA was identified in 1995 as a novel transcript increased in the striatum after acute cocaine or amphetamine administration. This transcript matched the protein sequence of an Ovine hypothalamic peptide partially sequenced by Spiess and Vale in 1981, implying that this transcript was indeed translated into an active peptide product. CART peptide is a multifunctional neuromodulator with roles in pain, reward, addiction, and food intake. Using our deductive reasoning strategy (Yosten et al. AJP303:R941, 2012) we have identified the previously orphaned G‐protein coupled receptor, GPR160, as a candidate CART receptor. Colocalization, proximity ligation, and immunoprecipitation studies have confirmed the association of CART and GPR160. We translated this evidence into in vivo rat food intake studies. The action of CART to inhibit food intake (4V administration) was lost in animals pre‐treated with siRNA targeting the GPR160 transcript. Additionally, passive immunoneutralization of GPR160 achieved by injecting antibody against the second extracellular loop of GPR160 into the fourth ventricle also blocked the anorexic effects of pharmacologic doses of fourth ventricle CART peptide. Taken together, these data identify GPR160 as a necessary component of the CART peptide signaling pathway and further demonstrate the requirement for this interaction to mediate the food intake reducing effects of CART peptide. Research on the function of CART peptide had been severely hindered by the lack of knowledge of a cognate receptor. The discovery of a CART peptide receptor will not only revitalize research in the CART field, but also potentially lead to novel therapeutic agents for the treatment of obesity and pain management.

Support or Funding Information

HL121456

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Modeling Effects of a High‐Fat (Almonds) vs. High‐Carbohydrate (Graham Crackers) Snack on Metabolic Outcomes in College Freshmen
Authors: Barron Marily , Dhillon Jaapna , Asghar Syed A. , Kuse Quintin , De La Cruz Natalie , Vu Emily , Sindi Suzanne S. , Ortiz Rudy M.
Published: June 2021

Biomedical research has provided support for the benefits of consuming almonds, a high‐fat food. However, most of the studies to date have considered only adult populations (age 40+). The objective of the study was to model the impact of consuming a high‐fat almond snack vs. a high‐carbohydrate graham cracker (control) snack for 4–8‐weeks in college freshmen. Seventy‐three UC Merced freshmen (Age: 18–19 years, BMI: 18–45 kg/m2) underwent a dietary intervention for eight weeks. Thirty‐eight participants consumed 2 ounces (325 kcal) of almonds and 35 participants consumed an isoenergetic control snack of graham crackers for 8 weeks. Anthropometric, biochemical and clinical measurements were conducted before the study began and at the fourth and the eighth week of the intervention. Several multivariate models were generated to model the effects of chronic almond and graham cracker snacking on the aforementioned outcomes. The most profound effect was the retention of high‐density lipoprotein (HDL) cholesterol and improvements in glucose tolerance in the almond snack group in comparison to the graham cracker snack group. This suggests that chronic almond snacking has the potential to improve the metabolic profile independent of profound changes in body mass or adiposity. We continue to perform the robust multivariate analyses factoring in BMI and sex effects.

Support or Funding Information

Almond Board of California, MARC U STAR, and UROC

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 Creatine Supplementation and Doxorubicin Treatment on Myogenic Regulatory Factor Expression in Slow and Fast Muscle
Authors: Torok Zoltan , Busekrus Raquel , Hydock David Scott
Published: June 2021

The purpose of this study was to investigate the effects of creatine monohydrate supplementation on myogenic regulatory factor expression in skeletal muscle from rats receiving the chemotherapy drug doxorubicin. Male Sprague‐Dawley rats were randomly assigned to the control (CON), doxorubicin (DOX), or creatine+doxorubicin (CR+DOX) groups. CR+DOX received rodent chow supplemented with 3% creatine monohydrate and the CON and DOX received standard rodent chow. After two weeks of feeding, CR+DOX and DOX groups received a bolus, 15 mg/kg intraperitoneal (i.p.) DOX injection and CON received an i.p. saline injection as a placebo. Dietary interventions then continued for five more days. The soleus and extensor digitorum longus (EDL) were excised five days post injection and expression of myogenic regulatory factors MyoD, Myf5, Mrf4, and myogenin was determined using Western blotting. In DOX solei, MyoD, Myf5, Mrf4, and myogenin expression was observed to be +25%, −10%, +5%, and −25%, respectively when compared to CON, and −6%, +11%, −17%, and −12%, respectively in CR+DOX solei when compared to CON. MyoD, Myf5, Mrf4, and myogenin expression in DOX EDL were observed to be −4%, −44%, −14%, and −29%, respectively when compared to CON and −24%, −58%, −11%, and −43%, respectively in the EDL from CR+DOX when compared to CON. In the soleus, a trend toward increased MyoD and Mrf4 expression and decreased Myf5 and myogenin expression was observed with DOX treatment. The trend of CR+DOX expressing increased soleus Myf5 expression when compared to both CON and DOX is of interest and should be explored further.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Attenuation of Vascular Contractility in Metastatic Breast Cancer Mice
Authors: Vorn Rany , Yoo Hae Young
Published: June 2021

Breast cancer (BC) is the one of crucial health burden among women worldwide. Even though the alteration of vascular function were observed by few researchers in metastatic BC, the changes in vascular reactivity and its underlying mechanism were not fully understood. Here, we investigate the physiological changes of vascular function from isolated mesenteric arteries in metastatic breast cancer mice model. In this study, 105 of 4T1 murine BC cells were injected into the subcutaneous or tail vein of female Balb‐c mice aged 8 weeks old. Mice were randomly divided into the following group: 1) control; n=10, 2) non‐metastatic model (4T1; n=10), 3) tumor‐bearing model (4T1M; n=10), and 4) tail vein injection model (4T1ML; n=10). Mesenteric arteries were removed after 4 weeks of tail vein injection group and 5 weeks of tumor‐bearing groups. Isometric tension was investigated in response to various vasoconstrictors such as phenylephrine (PhE), Serotonin (5‐HT), Angiotensin II (ANG II) and high potassium chloride (HK). Our results showed that PhE‐induced vasoconstriction was no significant differences in the non‐metastatic group compared to control. However, the attenuated vasoconstriction in response to PhE, 5‐HT, and High K was found in metastatic cancer groups. Application of L‐type voltage‐gated calcium channel inhibitor (Nifedipine) induced vascular relaxation with PhE‐induced contraction in each group, but there were no significant differences among groups. Interestingly, PhE‐induced contraction in metastatic groups was not inhibited by Rho kinase inhibitor (Y‐27632), but not in control. From above results, it is suggested that decreased Rho kinase activity might be attributable to attenuated vasoconstriction in mesenteric artery from advanced breast cancer mice.

Support or Funding Information

This study was supported by a National Research Foundation of Korea grant funded by Ministry of Science, ICT, & Future Planning (2015R1C1A1A01054038).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exercise characteristics in mitochondrial catalase overexpressing mice
Authors: Rao Shriya D. , Gaillard Charleston E. , McKinley Alexander J. , Klatt Benjamin T. , Shanely R. Andrew , Christian Brooke E. , Zwetsloot Kevin A.
Published: June 2021

High intensity or prolonged exercise can result in oxidative stress due to the generation of free radicals and reactive oxygen species, which in turn, can negatively impact skeletal muscle contractile function and exercise performance. Catalase, a naturally‐occurring antioxidant enzyme, converts hydrogen peroxide to water and oxygen (2H2O2 → 2H2O + O2). Mitochondrial‐targeted catalase transgenic (mCAT) mice have been studied within the context of disease pathology, but skeletal muscle function and exercise capacity have not been characterized. Therefore, the purpose of this study was to measure grip strength, exercise capacity, and exercise endurance in mCAT mice that overexpress catalase in the mitochondria of skeletal muscle and heart, among other tissues. Male and female mCAT and wild‐type (WT) littermate mice (3–8 mo old) performed 3 consecutive forelimb grip strength tests, with 1‐min rest between trials. Next, these mice performed a maximal exercise capacity test, which consisted of a 10‐min warm up at 10–12.5 m•min−1 and 5–10° incline, followed by 2‐min stages of increasing treadmill speed by 3 m•min−1 every stage; treadmill remained at 10° incline and mice ran to exhaustion. Then, 48 hours after the completion of the maximal exercise capacity test, all mice performed an exercise endurance test, consisting of a 10‐min warm up at 10–12.5 m•min−1 and 5–10° incline, followed by a run to exhaustion at a constant treadmill speed of 22 m•min−1 and incline of 10°. Grip strength, maximal exercise capacity, and exercise endurance were not different between mCAT and WT mice, respectively (p>0.05, all). These findings suggest that mitochondrial catalase overexpression does not alter grip strength, treadmill exercise capacity, or endurance in mice.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Conceptual Framework for the Core Concept of Mass Balance
Authors: Modell Harold I. , Michael Joel A.
Published: June 2021

The core concepts of physiology are descriptions of phenomena that have great explanatory power in every area of physiology. They are “big ideas” that are comprised of many smaller ideas. The process of “unpacking” a core concept yields a hierarchically arranged statement of the sub‐ideas that we have called a conceptual framework (CF). Our goal has been to produce CFs that will facilitate teaching and learning physiology.

We have unpacked three of the core concepts (homeostasis, flow down gradients, and cell‐cell communication) and validated the resulting conceptual frameworks with feedback from the physiology teaching community; this work has been reported in Advances in Physiology Education.

Here we present our conceptual framework for the core concept of mass balance (conservation of mass). This core concept (and its CF) describes the consequences of moving mass (liquids, gases, solids) into and out of defined compartments. These ideas are applicable at every level of organization in the organism.

We will solicit feedback to validate the framework and its applicability to various levels of physiology instruction.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Observable Differences in Symmetry and Volume of Jugular Foramen in Children with Craniosynostosis
Authors: Fourniquet Sophie , Mundinger Gerhard S. , Smith David , Mussell Jason C.
Published: June 2021

Craniosynostosis is the premature fusion of one or more calvarial sutures. Craniosynostosis can present as part of a genetic syndrome, as in Apert or Crouzon Syndrome, or it may be an isolated finding in non‐syndromic patients. The premature closure of these sutures can lead to morphologic and physiologic anomalies. Synostotic patients present with craniofacial malformations, increased intracranial pressure and intracranial hypertension. These patients often present with anomalous vascular and osseous anatomy, specifically, large emissary veins and stenotic cranial foramina.

CT scans were used to generate three‐dimensional models of the patients. The jugular foramen was manually modeled, using the intracranial foramen and the level of the odontoid process as anatomical landmarks. Each measurement was repeated three times to ensure consistency.

This case series serves as a basis for continued research into the anomalous anatomy of synostotic patients. When compared to their normal counterparts, synostotic patients present with markedly asymmetric jugular foramina. A high degree of variability was observed between patients, and bilaterally within each patient. Large differences in shape and volume are noted between the right and left foramen. At this point, due to the small size of the study, inferences about the patient population at large cannot be made. However, some general similarities were observed.

Our preliminary data show that in all subjects, the right jugular foramen was longer and wider than the left. It was noted that the right foramen has a more curved trajectory, moving in the posterior to anterior direction, while the left is relatively straight. This may be related to the left jugular foramen's apparent propensity to merge with the carotid canal.

The observed anomalies likely contribute to the physiologic sequelae of craniosynostosis. We hypothesize that the right jugular foramen is consistently larger due to the backwards pulse pressure coming from the right atrium of the developing fetus. We propose that the timing of the pathophysiologic insult determines the character of the osseous anomaly. Measuring the surface area two‐dimensionally leads to an incomplete and potentially inaccurate representation of the jugular foramen in synostotic patients due to the irregularity of caliber throughout the jugular foramen's course. It is proposed that this osseous irregularity is in part due to diminished post‐natal remodeling. We propose that the most appropriate and rigorous method for examining the jugular foramen in synostotic patients is by obtaining the volume of the jugular foramen by three‐dimensional modeling.

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 Squat Factor in Defense of a Vulnerable Neck and Brain
Authors: Naples Virginia , Rothschild Bruce
Published: June 2021

Pathological skeletons allow behavioral inferences for extant and extinct taxa without living analogs. Recent revisions of sauropod head, neck and tail posture suggest defensive use of their enormous tails to protect their hindquarters. Horizontal neck posture assigned to many of the sauropods, however, make the distal neck and head (brain) accessible to predation. Posterior neck proportions of even a small sauropod (e.g., 14 meter Camarasaurus) are 1.3 meters wide and 1.7 meters high, beyond the gape of contemporary predators. Anterior neck diameter (0.3 meters wide by 0.6 meters tall), however, is within the gape and accessible (vertically to large Allosaurus. Sauropod necks move maximally 20° laterally, eliminating defensive swinging away from a predator. Long bodies limit tail use during frontal attack. Some reconstructions show sauropods rearing on hind legs with anteriorly or anteroventrally extended forelegs. However, there is theoretical and physical evidence that they likely squatted with flexed knees as do other graviportal animals (e.g., elephants) in a stable, easily held posture. A unique diplodocid, KUVP 129717, with a partially healed, distal penetrating pubic symphysis injury, and a healed, deep anteroposterior dorsal claw cut metacarpal, provide clues to head and anterior neck defense. Slight remodeling, not altering injury architecture, proves the sauropod survived. The pubis wound is not explainable by tooth or claw injury and lacks the characteristic taper related to Stegosaurus spike‐derived injuries. The geometry and location suggest that the sauropod sat on a sharp, slender object, in a critical situation that prevented injury avoidance. Sauropods squatted to lay eggs, unlikely encountering a sharp object, or the animal would have repositioned to avoid pubic symphysis penetration. We suggest that the diplodocid squatted when imminent danger required anterior body elevation to raise the head and neck above predator reach. Xenarthran anteaters use this stable tripodal defensive stance, supporting partial body weight on the tail base as sauropods may have done. Further, presence of a long metacarpal claw‐derived scar suggests a defensive wound, one that could have occurred if it was elevated by squatting to the position that placed the distal neck and head out of reach of predator reach, leaving its forelimbs available for defense. Such a posture protects sauropods from even the largest Late Jurassic carnosaurs.

Support or Funding Information

Not applicable.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inner Ear Morphology of Basal‐Most Mammaliaform Morganucodon
Authors: Shahid Ramza , Gill Pamela G. , Hoffmann Simone
Published: June 2021

Extant therians are unique in their ability to hear at higher and at greater ranges of frequencies than most other vertebrates. This ability has been associated with changes in inner ear morphology, including elongation of the cochlear canal following loss of the lagena maculae, coiling of the cochlear canal, and stabilization of the hearing membrane through bony laminae. Presence and absence of these features is variable across Mesozoic mammaliaforms. To gain insights into the evolution of the inner ear and hearing in mammaliaforms, we document inner ear morphology in the basal‐most mammaliaform Morganucodon in unprecedented detail. We μCT scanned 30 petrosals of Morganucodon from several Jurassic fissure fillings in the UK. Surface reconstructions of the inner ears demonstrate that the cochlear canal gently varies in curvature and length (1.64–1.80 mm, measured from the posterior aspect of the cochlear foramen to the apex). All specimens show a distinct expansion of the apex of the cochlear canal suggesting the presence of a lagenar macula. Interestingly, the size of the apical expansion varies within our sample. None of the specimens preserve complete ossified laminae, but a shallow groove is visible on the ventrolateral surface of the cochlear canal endocasts, extending from the base of the canal (between the perilymphatic foramen and the fenestra vestibule) anteriorly towards the apex. We identify this groove as the attachment for the basilar membrane (termed here “base of secondary lamina”), as seen in extant monotremes. If the base of the secondary lamina is indeed reflective of basilar membrane length in Morganucodon then this could represents the first accurate reconstruction of the length of the basilar membrane in a fossil mammaliaform that retains a lagena. The base of the secondary lamina ends shortly before the apex of the canal and ranges between 1.17 and 1.45 mm. It moderately correlates (R2=0.58) with total cochlear canal length in our sample. Most intriguingly, the bony wall of the promontorium contains a network of canals that surrounds the cochlea, previously described as the circumpromontorium plexus. The plexus enters the cochlear canal next to the base of the secondary lamina and extends along the laminas full length. This strongly suggests that the blood supply to the cochlea might have been substantially different in Morganucodon compared to extant therians where blood vessel enter the cochlea through the cochlear foramen along with the cochlear nerve.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
HFE Genotype Restricts the Response to Paraquat in a Mouse Model of Neurotoxicity
Authors: Dower Anne Nixon , Neely Elizabeth , Meadowcroft Mark , Nandar Wint , Connor James
Published: June 2021

Parkinson's disease (PD) is marked clinically by motor dysfunction and pathologically by dopaminergic cell loss and iron accumulation in the substantia nigra (SN). The driver underlying iron accumulation is unknown and could be genetic or environmental. The HFE protein is critical for the regulation of cellular iron uptake. Mutations within this protein are associated with increased iron accumulation including in the brain. We have focused on the commonly occurring H63D variant of the HFE gene as a disease modifier in a number of neurodegenerative diseases. To investigate the role of H63D HFE genotype, we generated a mouse model in which the wild‐type (WT) HFE gene is replaced by the H67D gene variant (mouse homolog of the human H63D gene variant). Using paraquat toxicity as the model for Parkinson's disease, we found that WT mice responded as expected with significantly greater motor function, loss of tyrosine hydroxylase staining and increase microglial staining in the substantia nigra, and an increase in R2 relaxation rate within the substantia nigra of the paraquat‐treated mice compared to their saline‐treated counterparts. In contrast, the H67D mice showed a remarkable resistance to paraquat treatment; specifically differing from the WT mice with no changes in motor function or changes in R2 relaxation rates following paraquat exposure. At baseline, there were differences between the H67D HFE mice and WT mice in L‐ferritin staining in the substantia nigra that could account for the resistance to paraquat. Of particular note, the H67D HFE mice regardless of whether or not they were treated with paraquat had significantly less tyrosine hydroxylase immunostaining than WT. Our results clearly demonstrate that the HFE genotype impacts the expression of tyrosine hydroxylase in the substantia nigra and the response to paraquat providing additional support that the HFE genotype is a disease modifier for PD. Moreover, the finding that the HFE mutant mice are resistant to paraquat may provide a model in which to study resistant mechanisms to neurotoxicants.

Support or Funding Information

The work is supported in part by the Pennsylvania Department of Health Tobacco CURE Funds and the Penn State Neuroscience Institute.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mind Over Matter: Understanding the Relationship Between Memory Self‐Efficacy, Cognition and Brain Health in Older Adults with Probable Mild Cognitive Impairment
Authors: Horst Rebecca , Nagamatsu Lindsay S.
Published: June 2021

Background

In our aging population, cognitive decline and brain health are critical areas of concern for healthy aging. Evidence has shown that personality factors such as self‐efficacy, one's personal perceived ability to perform a specific task, directly impacts components of healthy aging, including total brain volume. However, it is unknown whether memory self‐efficacy, specifically, might also be associated with brain function and structure.

Methods

A cross‐sectional pilot study of community dwelling older women with probable Mild Cognitive Impairment (Montreal Cognitive Assessment score <26) were asked to evaluate their global memory self‐efficacy using two questionnaires: Memory Self‐Efficacy Questionnaire (MSEQ‐4) and the Multifactorial Memory Questionnaire (MMQ). Participants were asked to complete various standardized cognitive tests: Alzheimer's Dementia Assessment Scale – Cognition (ADAS‐cog), Digit Span, Auditory Verbal Learning Test, Stroop and Trail Making Test. Participants also performed an associative memory task during an fMRI scan. High resolution T1 weighted structural imaging was obtained from a 3T SIEMENS scanner. Multivariate linear regression models were constructed for cognitive and brain health measures in relation to the memory self‐efficacy measures. Covariates of the models included age and current physical activity level.

Results

We report that the MMQ subscale of Mistakes and Ability (MMQ‐A) was the strongest measure in accounting for variance after including covariates. The final model for ADAS‐cog accounted for 65% of the variance, with the MMQ‐A score accounting for 44%. For structural brain measures, total brain volume, white matter and grey matter volumes, the final model accounted for 70%, 98% and 13% for each of the listed measures, with MMQ‐A accounting for 52%, 63% and 9% respectively. Other measures of global memory self‐efficacy, MSEQ‐4 and MMQ subscale of feelings of contentment (MMQ‐C), were also seen to have correlations to ADAS scores and structural brain measures, but could not account for the same level of variance as the MMQ‐A.

Conclusion

Based on the results collected it appears that one's perceived self‐efficacy of memory mistakes and ability is associated with measures of cognition and brain health. Based on this data our research has the potential to progress into a longitudinal study of observing the relationship between changes in memory self‐efficacy and brain health and cognition, as well as progression to collaborative clinical studies in memory self‐efficacy modification for healthy aging.

Support or Funding Information

Funding: Natural Sciences and Engineering Research Council of Canada

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Development of mouse superior cervical ganglion during prenatal and postnatal stages.
Authors: Sato Iwao , Mitsuoka Kazuyuki , Miwa Yoko
Published: June 2021

The neuropeptide calcitonin gene‐related peptide (CGRP) is a well‐characterized mediator of receptor‐regulated neurotransmitter release. However, little is known between CGRP during the development of the superior cervical ganglion (SCG). In the present study, we observed the localization of the CGRP mRNA using in situ hybridization during development from the embryonic stage to after birth day 14.5 (E14.5), E17.5 and postnatal day 1 (P1). The different ratio was found in SCG ganglion cells and shrunken cells. The antisense probe for CGRP was detected by in situ hybridization at E14.5, E17.5, and P1, and the highest levels of positive CGRP ganglion cells were detected at E17.5. The specific characters of CGRP concentration and release were occurred during development. The CGRP may influence the functional stress during prenatal and postnatal development.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Assessing the Effects of Inorganic Arsenic on IL‐1β and TNFɑ Secretion, Gene Expression, and DNA Methylation in Murine Macrophages to Gauge Immunotoxic Effects of Heavy Metals
Authors: DeLong Lauren Nicole , Barbosa Jeremie Rose , Nyland Jennifer
Published: June 2021

Trivalent inorganic arsenic, arsenite, is a natural component of the earth and is often found in drinking water in areas where the composition is high. Some studies have suggested that, especially at moderate to high exposure levels, arsenite is toxic to individuals who are chronically exposed through drinking water, characterized by an increase in inflammatory cytokine release, specifically interleukin‐1β (IL‐1β) and tumor necrosis factor‐ɑ (TNFɑ). Other epidemiological studies at low to moderate exposures did not find the same correlation with inflammation. We aim to characterize and quantify the effects of arsenite exposure to macrophages to gauge any potential inflammatory response.

M1 macrophages are key players in inflammatory response and secrete IL‐1β and TNFɑ in response to Danger Associated Molecular Patterns (DAMPs) released by neighboring cells undergoing necrosis, in coordination with Pathogen Associated Molecular Patterns (PAMPs), molecules associated with pathogens, such as lipopolysaccharide, or LPS. We obtained a murine macrophage cell line to study the effects of arsenic on inflammatory cytokine secretion in vitro. A dose response curve was developed including varying concentrations of arsenite with and without 50 ng/mL LPS, using the same treatments without arsenite as controls. Cytolethality was determined by differential staining and counting. Secretion of cytokines after 24 hours of exposure was quantified using ELISA. Results were statistically analyzed using Student's t‐test and One‐Way ANOVA. IL‐1β was not differentially secreted for any concentration of arsenite, but cytolethality and TNFɑ secretion were significantly increased for 200 nM arsenite with LPS as opposed to 200 nM arsenite without LPS, possibly indicating that some inflammatory augmentation occurs in the presence of both arsenite exposure and a PAMP.

Future steps include analyzing alterations in expression of genes responsible for proteins involved in the secretory pathways of these cytokines. For example, the genes involved in manufacturing the pieces of the NLRP3 inflammasome, key in production of IL‐1β, or TNFɑ Converting Enzyme, involved in cleavage of the TNFɑ precursor, may be potential targets. Additionally, we will explore differential DNA methylation using methylation sensitive and insensitive restriction enzyme digestion to examine epigenetic control of cytokine secretion in response to arsenite. These results will be reported to appraise the immunotoxic effects of arsenic exposure.

Support or Funding Information

This research is supported by the Salisbury University Department of Biological Sciences and the Salisbury University Honors College.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Fas‐associated factor 1 (FAF1) binds to mineralocorticoid Receptor (MR) through its SUMO‐interacting motifs and negatively modulates MR transcriptional activity
Authors: Lin Ding‐Yen
Published: June 2021

Fas‐associated factor 1 (FAF1) was originally isolated as a Fas(CD95)‐associated protein factor and was subsequently found to interact with numerous other proteins that are involved in various cellular events including Fas‐mediated apoptosis, NF‐kB, Wnt/β‐catenin pathways, mineralocorticoid receptor (MR) mediated transactivation and ubiquitin‐dependent processes. Here, we define two Small Ubiquitin‐like Modifier (SUMO)‐interacting motifs (SIMs) within FAF1 and show it to be crucial for transcriptional modulation of MR. Our study demonstrated that the SIMs of FAF1 do not play a significant role in the regulation of its subcellular localization, Fas‐mediated apoptosis, NF‐kB and Wnt/β‐catenin pathways. Remarkably, FAF1 interacts sumoylated MR and represses aldosterone‐activated MR transactivation in a SIMs‐dependent manner. Moreover, silencing of endogenous FAF1 in stably MR‐expressing cells resulted in an increase in the induction of MR target genes by aldosterone, indicating that FAF1 functions as a MR corepressor. We further provide evidence to suggest that the mechanism of FAF1/SIMs‐mediated MR transrepression involves inhibition of MR N/C interaction. Sumoylation has been linked to impart repressive properties on several transcription factors and cofactors. Our findings therefore provide the mechanistic insight underlying SUMO‐dependent transcriptional repression of the MR.

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