bluuebunny/crossref_metadata_2025 huggingface dataset:
Atorvastatin does not ameliorate lithium‐induced nephrogenic diabetes insipidus in mice
Authors: Markussen Maria Linaa , Iervolino Anna , Rej Soham , Trepiccione Francesco , Christensen Birgitte Mønster
Published: June 2021

Lithium (Li) is an important mood‐stabilizing drug used in the treatment of affective disorders. However, as a significant adverse effect patients can develop Nephrogenic Diabetes Insipidus (NDI). NDI is characterized by the inability of the kidney to concentrate urine in response to vasopressin, resulting in severe polyuria and an increased long‐term risk of chronic kidney disease. Li‐NDI is associated with downregulation of the water channel AQP2 and a cellular remodeling of the kidney collecting duct. Effective and safe treatment of Li‐NDI is still lacking. Statins are cholesterol‐lowering drugs, which appear to increase AQP2 membrane‐translocation and improve urine concentration ability in other NDI models.

This project will investigate if atorvastatin is able to prevent the Li‐induced changes in mice using a mouse cortical collecting duct cell line (mCCD) and C57BL/6 mice.

Biotinylation assays showed that acute (1hr) treatment with simvastatin (p<0.05) or fluvastatin (p<0.0001) increased AQP2 membrane accumulation compared to controls in the mCCD cells. This confirms that the mCCD cell line respond to acute statin treatment as shown before for these two statins in other cellular and animal models. In order to see whether chronic statin treatment abolish the effects of Li, cells were treated with Li and Li/atorvastatin for 48 hrs. Li reduced total AQP2 levels (p<0.01) compared to controls as expected, but combined Li/atorvastatin treatment did not prevent the Li‐induced downregulation of AQP2. Chronic (21 days) Li treatment of mice increased urine output (p<0.0001) and reduced urine osmolality (p<0.0001) compared to controls (receiving normal food). Atorvastatin (given in combination with Li for 21 days) decreased levels of triglycerides in the blood (p<0.05) compared to controls. Combined Li/atorvastatin treatment did not prevent the effects of Li on urine output and urine osmolality. In kidney inner medulla, western blotting showed that Li reduced total AQP2 levels (p<0.001) and increased pS261‐AQP2 levels (p<0.05), and further showed a tendency to pERK1/2 upregulation (p=0.06) compared to controls as shown previously. However, combined Li/atorvastatin treatment did not abolish any of these changes. Immunohistochemistry further showed that atorvastatin did not prevent the Li‐induced increase in intercalated cells and increased proliferation in the inner medulla.

In conclusion, both chronic in vitro and in vivo experiments showed that atorvastatin does not appear to have positive effects on prevention of Li‐NDI in mice.

Support or Funding Information

Danish Medical Research Council, Beckett Fonden, Aase and Ejnar Danielsens Fonden, Knud and Edith Eriksens Mindefond

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Role of adrenomedullin in mediating water reabsorption in rat inner medullary collecting ducts
Authors: Wang Yanhua , Ma Fuying , Klein Janet D
Published: June 2021

Adrenomedullin (ADM) increases water permeability in the inner medullary collecting duct by mediating aquaporin‐2 (AQP2) phosphorylation. It is generally considered to be a backup system for vasopressin. We investigated whether adrenomedullin stimulates osmotic water permeability through cAMP and/or protein kinase A (PKA). RNA sequencing and western analysis showed that ADM and its receptors, calcitonin‐receptor‐like receptor (CRLR) and receptor‐activity‐modifying proteins 2 or 3 (RAMP2 or RAMP3) are highly expressed in the inner medullary collecting ducts of control rats. No mRNA expression was seen for RAMP1. We measured water permeability in inner medullary collecting ducts of rats by tubule perfusion with and without treatment with the PDE4 inhibitor roflumilast or the PKA inhibitor H89. ADM significantly increased osmotic water permeability from 251.4 ± 58.1 to 383.4 ± 73.2 μm/s (n=4, p<0.05). Roflumilast (30nM) further increased ADM‐stimulated osmotic water permeability to 472.7 ± 84.9 μm/s (n=4, p<0.05). Inhibition of PKA by H89 decreased ADM‐stimulated osmotic water permeability from 225.4 ± 57.3 to 179.7 ± 49.6 μm/s (n=3, p<0.05). Western analysis with phospho‐specific antibody showed that ADM significantly increased phosphorylation of AQP2 at serine 256 and decreased phosphorylation of AQP2 at serine 261. We conclude that ADM and its receptors are expressed in inner medulla and that the mechanism of ADM stimulation of water reabsorption involves cAMP and activation of PKA. Further, our results suggest that ADM may represent a complimentary method for vasopressin in regulating water homeostasis.

Support or Funding Information

This work was supported by Emory URC grant 00067457, and AHA Career Development Grant #18CDA34060053.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
TRPC6 deficiency causes obesity and metabolic dysfunction
Authors: Bailey Kandice , Wang Zhen , Moak Sydney P. , Dai Xuemei , Fu Yiling , do Carmo Jussara M. , da Silva Alexandre A. , Hall John E.
Published: June 2021

Transient receptor potential cation channel C (TRPC) is a group of receptor‐operated cation channels that modulate cell Ca2+ influx. TPRC6 is a subunit of the TRPC family that is expressed in the brain. However, the role of TRPC6 in controlling metabolic and cardiovascular functions remains unknown. In this study, we investigated the impact of genetic TRPC6 deletion on energy balance, metabolic and cardiovascular functions, and anorexic responses to leptin. Metabolic phenotypes including body weight (BW), food intake (FI), body fat/lean composition, energy expenditure (EE), respiratory quotient (RQ), responses to acute leptin injection as well as cardiovascular parameters including blood pressure (BP) and heart rate (HR) were measured by telemetry in male and female TRPC6 null and B6/129 control mice (n=7/per group and sex). TRPC6 null mice were heavier than control mice from 6 to 16 weeks of age when fed a standard diet (39.5±1.5 vs 31.1±1.2 g in males and 31.1±1.2 vs 22.9±1.0 g in females at 16 weeks of age, p<0.05). EchoMRI scans showed that the higher BW of TRPC6 null mice was mainly due to increased body fat compared to controls (30.0 vs 22.6 % of fat mass/BW in males and 32.7 vs 21.0 % in females at 16 weeks of age), and was associated with increased FI (3.4±0.1 vs 2.7±0.2 g/day in males and 2.7±0.1 vs 2.0±0.1 g/day in females at 16 weeks of age, p<0.05). EE, assessed by indirect calorimetry, was significantly reduced in TRPC6 null mice (0.108±0.003 vs 0.205±0.012 kcal/12h/g in males and 0.178±0.005 vs 0.217±0.006 kcal/12h/g in females at 17 weeks of age, p<0.05) and was accompanied by significantly reduced 24‐h average RQ (0.80±0.01 vs 0.89±0.01 in males and 0.77±0.01 vs 0.84± 0.01 in females at 17 weeks of age, p<0.05) when compared to male and female controls. Acute leptin injections (5 mg/kg, i.p. at 18 weeks of age) reduced 24‐hr FI by 41% and 32 % in male and female control mice, respectively, while only 8 % and 18 % reductions in FI were observed in male and female TRPC6 null mice. BP and HR measured by telemetry for 5 consecutive days at 22 weeks of age were similar in TRPC6 null mice and control mice (MAP: 111±2 vs 108±2 in males and 103±3 vs 108±3 in females; HR: 541±9 vs 526±14 in males and 552±4 vs 552±27 in females). Our results indicate that TRPC6 plays an important role in normal control of FI, BW and energy balance as well as for normal anorexic responses to leptin. Although TRPC6 deficiency caused obesity and metabolic abnormalities, BP and HR did not increase suggesting that TRPC6 may be an important link between obesity and hypertension.

Support or Funding Information

NHLBI PO1HL51971, NIGMS P20GM104357 and U54GM115428, and NIDDK R00DK113280

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
New Methods for Analysis and Interpretation of Obesity Experiments
Authors: Banks Alexander , Corrigan June
Published: June 2021

We report here the first data repository for experiments of obesity and body weight change. Modern obesity research depends upon indirect calorimetry systems to allow for state‐of‐the‐art experimental measurement of energy expenditure and food intake. These systems generate a flood of raw data that require time‐consuming manual manipulation for formatting, outlier detection, statistical interpretation, and visualization. Analysis of calorimetry experiments requires an onerous and specialized statistical treatment to account for differences in body mass and body composition. As such, controversies have emerged on the appropriate treatment of the data generated by these experiments, fundamentally challenging some published conclusions. We recently described a free web‐based tool, CalR, for analysis of experiments using indirect calorimetry to measure physiological energy balance. CalR simplifies the process to import raw data files, generate plots, and determine the most appropriate statistical tests for interpretation of diverse experimental designs including those of obesity and thermogenesis. Critically, CalR can read in data files from all experimental setups and produce a standardized output file.

A recent effort to promote transparency and increase the rigor of the scientific process, especially regarding biostatistical analysis and improving reproducibility, has created an atmosphere receptive towards novel tools that assist in achieving these goals. Many forms of large scale datasets have central repositories, including that for microarray, RNA‐seq, and proteomics experiments. Here describe the first repository for indirect calorimetry experiments, populated by reference data from more than 30,000 individual mice from the International Mouse Phenotype Consortium (IMPC). By uploading data, in the standard CalR format to the repository, users can see how their experiments compare to the reference data. Furthermore, the user‐supplied data can be shared with the public as a lasting resource. This new system will provide the transparency necessary to enhance consistency, rigor, and reproducibility in studies of obesity and energy balance.

Support or Funding Information

‘Financial support for this work was provided by the NIDDK Mouse Metabolic Phenotyping Centers (MMPC, www.mmpc.org) under the MICROMouse Program, grants DK076169 and DK115255.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Changes in Adipose Tissue Physiology during the First Two Weeks Post‐hatch in Chicks from Lines Selected for Low or High Body Weight
Authors: Xiao Yang , Siegel Paul B. , Cline Mark A. , Gilbert Elizabeth R.
Published: June 2021

Chickens from lines selected for low or high body weight from a common foundation population differ in appetite and adiposity; however, mechanisms associated with the predisposition to becoming obese are unclear. The objective of this study was thus to evaluate developmental changes in depot‐specific adipose tissue during the first 14 days post‐hatch. The subcutaneous (SQ), clavicular (CL) and abdominal (AB) adipose depots were collected for histological and mRNA expression measurements from chicks of each line on day of hatch, day 4 and day 14 post‐hatch. Results showed that low line chicks had decreased SQ fat mass on a body weight basis with reduced adipocyte size from day of hatch to day 4 and increased body weight and fat mass with unchanged adipocyte size from day 4 to day 14 post‐hatch. High line chicks had increased body weight throughout the first 14 days post‐hatch, and increased fat mass in all three depots with significantly enlarged adipocytes in AB from day 4 to day 14 post‐hatch. Meanwhile, mRNA expression of CCAAT/enhancer‐binding protein alpha, neuropeptide Y and peroxisome proliferator‐activated receptor gamma, which are involved in adipogenesis, as well as acyl‐CoA dehydrogenase, long chain, which catalyzes fatty acid oxidation, had different expression patterns among depots between lines at different ages. From day of hatch to day 4 post‐hatch, low line chicks mobilized SQ fat and replenished the reservoir mainly through hyperplasia, whereas in the high line there was dependency on both hyperplasia and hypertrophy to maintain adipocyte size and depot mass. From day 4 to day 14, adipose tissue catabolism and adipogenesis slowed down in both lines. While the fat depot mass and adipocyte size were stable in the low line, there was rapid accumulation of fat in CL and AB regions of high line. Chicks predisposed to be anorexic or obese display different fat development patterns during the early post‐hatch period.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Daily Rhythms Are Altered by High‐Fat Feeding in Female Offspring Born to Obese Mothers
Authors: Llanora Josie Dianne , Pendergast Julie S
Published: June 2021

About 20% of women are obese during pregnancy. Children born to obese mothers are at increased risk of obesity, impaired glucose tolerance, and other facets of metabolic syndrome. Disruption of circadian rhythms is also associated with obesity and metabolic dysfunction. Circadian, or daily, rhythms are approximately 24‐hour fluctuations in physiology and behavior. Environmental factors, such as the light‐dark cycle and diet, can alter these rhythms. For example, in male mice, high‐fat diet (HFD) feeding alters daily rhythms and increases body weight. In contrast, female mice fed HFD do not become obese, and their daily rhythms are not altered. Previous studies have shown that female mice will develop diet‐induced obesity if they are born to an obese mother. In this study, we sought to determine if female offspring weaned from obese mothers had disrupted daily rhythms of eating behavior and locomotor activity. Female C57BL/6J mice were fed HFD (60% kcal fat) for 12 weeks (6 weeks prior to pregnancy and throughout pregnancy and nursing). After weaning, female offspring were fed low‐fat diet (LFD, 10% kcal fat) for 5 weeks and then HFD for 8 weeks. We first continuously measured daily eating behavior rhythms with infrared video cameras. All female offspring, regardless of whether the mother was lean or obese, had high‐amplitude rhythms of eating behavior during LFD feeding. Consistent with our previous studies, HFD consumption did not alter the eating behavior rhythms of females born to lean mothers. In contrast, HFD consumption reduced the amplitudes, or robustness, of the eating behavior rhythms in females born to obese mothers. We also investigated the effects of HFD on locomotor activity rhythms, measure by infrared sensors, in female offspring born to lean and obese mothers. The daily rhythms of activity were not affected by HFD feeding in females born to lean mothers. However, the phases (or timing) of the activity rhythms were altered by HFD feeding, compared to LFD feeding, in females born to obese mothers. Together, our results show that maternal obesity causes daily rhythms in female offspring to be susceptible to the effects of HFD feeding.

Support or Funding Information

This study was funded by an American Physiological Society Undergraduate Summer Research Fellowship, the Gertude F. Ribble Trust, and the University of Kentucky.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Splenectomy Increases Blood Pressure and Alters the Renal T Cell Profile in a Sex‐Specific Manner in Spontaneously Hypertensive Rats
Authors: Gillis Ellen E. , Sullivan Jennifer C.
Published: June 2021

Over the past decade there has been increasing evidence for the role of the immune system in the development of hypertension. Our lab has previously reported that female Spontaneously Hypertensive Rats (SHR) have a blood pressure (BP)‐dependent increase in anti‐inflammatory regulatory T cells (Tregs), corresponding to lower BP compared to males. Based on the central role for immune cells in the control of BP and the known sex differences in the immune cell profile, the current study was designed to test the hypothesis that removal of the spleen will have a sex‐specific impact on BP in SHR. To test this hypothesis, baseline systolic BP was measured by tail cuff plethysmyography in male and female SHR. Rats were randomized to receive sham surgery (CON) or splenectomy (SPLNX, n=3). After 2 weeks, BP was recorded again and kidneys were harvested for flow cytometric analysis of T cells. BP was significantly higher in males compared to females at baseline and at Week 2 (Table, Effect of sex, p=0.0002 and p=0.0001, respectively, 2‐Way ANOVA). Splenectomy increased BP in both sexes after 2 weeks (Effect of SPLNX, p=0.0008), with females having a greater increase in BP than males (Interaction effect, p=0.05). Splenectomy did not alter total CD4+ cells in the kidney, but did affect the subpopulations of CD4+ cells. Renal Tregs (CD3+ CD4+ FoxP3+ cells, expressed as % CD3+ CD4+) decreased in both sexes after splenectomy, abolishing the sex differences in renal Tregs (Table, Effect of SPLNX, p<0.001, Interaction effect, p=0.0015). Renal pro‐inflammatory Th17 cells (CD3+ CD4+ RORγ+ cells, expressed as % CD3+ CD4+) increased in both sexes with splenectomy (Table, Effect of SPLNX, p<0.001). In conclusion, removing the spleen in male and female SHR significantly increased BP, with females having a greater increase in BP compared to males. Interestingly, this greater increase in BP in the female was accompanied by a greater reduction in renal Tregs, supporting the idea that renal Tregs play a critical role in BP control in female SHR, contributing to the sex difference in BP. Ongoing studies in the laboratory are further exploring the mechanisms for the relative increase in renal Tregs in females.

Support or Funding Information

This work was funded by AHA18POST34030252 to EEG and 5R01HL127091 to JCS.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Deletion of NHE3 Selectively in The Proximal Tubules Does not Alter the Blood Pressure Response to Angiotensin III in Male and Female Proximal Tubule‐Specific NHE3 Knockout Mice
Authors: Chen Xu , Li Xiao C , Zheng Xiaowen , Zhao Chunling , Soleimani Manoocher , Zhuo Jia L
Published: June 2021

Objectives

We have recently shown that the pressor and renal Na+ excretory responses to angiotensin II (ANG II) were significantly attenuated in global (Nhe3−/−), kidney‐selective (tgNhe3−/−), and proximal tubule‐specific NHE3 knockout mice (PT‐Nhe3−/−). The present study tested the hypothesis that deletion of NHE3 selectively in the proximal tubules of the kidney alters the blood pressure and renal Na+ responses to its major metabolite, ANG III, which reportedly binds and activates AT2 receptors and inhibits NHE3 in the proximal tubules.

Methods

Proximal tubule NHE3 knockout mice (PT‐Nhe3−/−) were generated using the sglt2‐Cre/NHE3‐floxed approach. To test the hypothesis, adult and age‐matched male and female wild‐type littlemates (WT) and PT‐Nhe3−/− mice (n = 10–11 each group) were infused with or without ANG III at 1.5 mg/kg/day via osmotic minipump, i.p., for 2 weeks. Systolic, diastolic and mean arterial blood pressure, and 24 h urinary Na+ excretory responses were determined before and weekly after treatment.

Results

Unlike global Nhe3−/− mice, no abnormal intestinal structural and absorptive dysfunction was found in proximal tubule‐specific NHE3 knockout mice (PT‐Nhe3−/−). At the basal level, systolic, diastolic, and mean arterial blood pressure was significantly lower (p<0.01), whereas 24 h urine and urinary Na+ excretion were significantly higher in male PT‐Nhe3−/− than male WT mice (p<0.01). ANG III significantly increased systolic blood pressure from 113 ± 2 mmHg to 127 ± 3 mmHg in WT mice (n=10, p<0.01 vs. baseline), and from 104 ± 3 mmHg to 116 ± 5 mmHg in PT‐Nhe3−/− mice (n=11, p<0.01 vs. baseline). However, there were no differences in the net pressor and heart rate responses to ANG III between male and female WT and PT‐Nhe3−/− mice. ANG III also significantly increased 24 h urinary Na+ excretion in male WT mice (n=10, p<0.05 vs. baseline), but not in male PT‐Nhe3−/− mice (n=11, n.s.). There were no significant differences in 24 h urinary Na+ excretion between female WT (n=10, n.s.) and PT‐Nhe3−/− mice (n=10, n.s.). Finally, ANG III infusion did not significantly alter the heart‐ or kidney‐to‐body wt. ratio, blood volume, or hematocrit in male and female WT and PT‐Nhe3−/− mice, respectively.

Conclusion

The deletion of NHE3 selectively in the proximal tubules of the kidney doesn't significantly alter the blood pressure responses to ANG III in male and female PT‐Nhe3−/− mice, but it does attenuate the natriuretic response to ANG III in male PT‐Nhe3−/− mice.

Support or Funding Information

This work was supported in part by NIH grants, 2R01DK102429‐03A1, 2R01DK067299‐10A1, and 1R56HL130988‐01 to Dr. Jia Zhuo.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
TLR4‐driven Lymphatic Alterations during IBD are driven in PAMP/DAMP Specific Manners
Authors: Stephens Matthew , von der Weid Pierre‐Yves
Published: June 2021

Background

Inflammatory bowel disease (IBD) is characterised by both acute and chronic phase inflammation of the gastro‐intestinal (GI) tract that affect a large and growing number of people worldwide with little to no effective treatments. This is in part due to the lack of understanding of the disease pathogenesis and also the currently poorly described involvement of other systems such as the lymphatics. During DSS induced colitis, mice also develop a severe inflammation of both the colon and terminal ileum with many features similar to IBD. As well as inflammation within the intestinal tract we have previously demonstrated lymphatic remodelling within the mesentery and mesenteric lymph nodes of DSS‐treated mice. The lymphatic remodelling includes lymphangiogenesis, lymphatic vessel dilation and leakiness, as well as cellular infiltration into the surrounding tissue and peripheral draining lymph nodes. The aim of the investigation was to delineate the contribution of PAMP‐ and/or DAMP‐driven lymphatic remodelling and inflammation during disease, in respect to Toll‐like receptor 4 (TLR4) activity.

Methods

Intestinal inflammation was induced in C57BL/6 mice by administration of 2.5% DSS in drinking water for 7 days. Mice were treated with TLR4 blocker C34 or Polymyxin‐B (PMXB) daily from days 3–7 of DSS treatment via I.P. injection, and their effects on disease activity and lymphatic function were examined. TLR4 activity and subsequent effect on lymphangiogenesis, lymphadenopathy, and mesenteric lymph node (MLN) cellular composition were assessed.

Results

DSS Mice treated with TLR4 inhibitor, C34, had a significantly improved disease phenotype characterised by reduced ileal and colonic insult. The change correlated with significant reduction in colonic and mesenteric inflammation, resolved mesenteric lymphangiectasia, and CD103+ DC accumulation similar to that of healthy control. PMXB treatment did not resolve inflammation within the colon, or associated mesenteric lymphatic dysfunction, but did however prevent lymphadenopathy within the MLN through alteration of CCL21 gradients and CD103+DC migration.

Conclusions

TLR4 appears to mediate several changes within the mesenteric lymphatics, more specifically it is shown to have different outcomes whether stimulation occurs through pathogen derived factors such as LPS or tissue derived DAMPs, a novel phenomenon.

Support or Funding Information

This study was supported by the Lymphedema Research and Education Program, Snyder Institute for Chronic Diseases, Cumming School of Medicine, University of Calgary to P.‐Y.v.d.W.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Translational Control of FGF21 mRNA Expression is Responsive to Nutritional Stress
Authors: Welles Jaclyn E. , Jefferson Leonard S. , Kimball Scot R.
Published: June 2021

Fibroblast growth factor 21 (FGF21) is a peptide hormone associated with the pathogenesis of several metabolic disorders such as Non‐alcoholic Fatty Liver Disease (NAFLD), Type‐2‐Diabetes, and obesity. Notably, FGF21 treatment is associated with enhanced insulin sensitivity, and in rodents, attenuation of the metabolic defects associated with consumption of a high‐fat diet. Because the majority of circulating FGF21 is secreted by the liver in mammals, understanding the mechanisms that regulate hepatic expression of the hormone is critical. Hepatic FGF21 expression has been shown to be regulated in part through changes in FGF21 gene transcription. However, recent studies have shown transcription‐independent changes in FGF21 secretion from isolated hepatocytes, implying that FGF21 protein expression is also regulated at the level of mRNA translation. Thus, the goal of the present study was to assess whether or not the FGF21 mRNA is translationally regulated using H4IIE hepatoma cells as a model system. In these studies, cells were treated with rapamycin as a mimic of starvation and translation of the FGF21 mRNA was assessed using sucrose density gradient centrifugation to separate mRNAs that are poorly translated, i.e. mRNAs associated with 3 or fewer ribosomes (“light” polysomes) from well‐translated mRNAs, i.e. mRNAs associated with four or more ribosomes (“heavy” polysomes). As a positive control, mRNAs bearing a terminal oligopyrimidine tract (TOP) at the 5′‐end of the message were also assessed, as previous studies have shown them to be translationally downregulated under the conditions used in this study. As expected rapamycin treatment resulted in repressed translation of the TOP mRNAs, as demonstrated by a shift of the mRNAs from heavy to light polysomes. In contrast, translation of the FGF21 mRNA was enhanced by rapamycin, suggesting that it is regulated through an mTORC1‐dependent mechanism distinct from, and opposite to, mRNAs with a TOP motif. Current efforts are directed toward identifying the regulatory element(s) in the FGF21 mRNA that engenders sensitivity to rapamycin‐induced translational upregulation.

Support or Funding Information

(Supported by NIH grant DK13499)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Reversal of Hepatomegaly Following Cessation of Kratom Consumption in C57BL/6 Male and Female Mice
Authors: Guenther Emily , Musick Maggie , Davis Tracy
Published: June 2021

Kratom is derived from the leaves of a Southeast Asian plant, Mitragyna speciosa. Mitragynine is one of several compounds in kratom, but has been shown to bind to the opioid receptors. Numerous people in the United States take kratom to alleviate pain, help with depression, and as a substitute during opioid withdrawal. Little is known about the effects kratom has on various organ systems. The objective of the current study was to determine if hepatomegaly caused by 10 days of consuming kratom can be reversed after 30 days of discontinuance of kratom. A kratom tea was made by placing the kratom powder in a tea bag and boiling for 10 min in spring water that was brought to a pH 4 with fresh lemon juice. Mice received a liquid diet for 3 days of acclimation. Beginning on day 6, mice received the liquid diet (control) or the liquid diet made with kratom tea with various doses of mitragynine (0.175 mg/kg BW−1) as determined by HPLC. These doses were based on allometric scaling to approximate human doses. Mice were paired fed for 10 days and tissues collected day 11 from half the mice. The remaining mice were weaned back to their pellet diet and tissues were collected 4 weeks later. Blood was collected via heart puncture and serum stored at −20°C to be analyzed for liver enzyme function and mitragynine concentration. Kidneys and small intestines were collected for analyses in future studies. Livers were collected, wet weight recorded, and portions were snap frozen in liquid nitrogen for RNA analysis and embedded in OCT or fixed in 10% buffered formalin for histology. Consumption of daily large doses of kratom appeared to have no outward health issues when the daily activities of the mice were observed. Liver size was greater (P < 0.01) on day 11 in mice consuming kratom (4.42 ± 0.31% of body weight; n = 10) as compared to controls (6.52 ± 0.31% of body weight; n = 6). At 4 weeks following cessation of kratom, liver size was no different in kratom‐treated (4.88 ± 0.44% of body weight; n = 6) as compared to control mice (4.51 ± 0.27% of body weight; n = 10). Adhesions of the liver to the diaphragm, stomach, right kidney, or small intestine were observed at 4 weeks in mice that received kratom. Alterations to enzymes or inflammatory cytokines in liver tissue have not yet been evaluated. Consumption of kratom in as few as 10 days causes hepatomegaly that can be reversed if kratom is discontinued. The extent of liver damage and dysfunction and effects on future health status has yet to be determined.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Creating a Course in Science Teaching to Address the Needs of Graduate Trainees
Authors: Downey Ryan M , Mulroney Susan E , Myers Adam K
Published: June 2021

Graduate students and post‐doctoral fellows who wish to pursue academic careers have a desire and need for formal instruction to become effective science educators. To meet this need within our own academic programs, a new course was created for the fall of 2018: Learning Design for Science Education. The primary goals of this course were to introduce students to fundamental theories of science teaching and to provide students with hands‐on experience with planning, administering, and teaching a science course. Our course was divided into two components: 1) a 9‐week instructional block in which foundational pedagogical theory was introduced and current best practices in active learning, feedback, and evaluation were role modeled by course faculty; and 2) a 4‐week practical application block in which student groups designed and conducted a mini‐course incorporating active learning methods and modern teaching technologies. The final week of the course was dedicated to providing detailed debriefings of the mini‐courses. Throughout the semester, students created a portfolio of their work, including a personal teaching statement, course syllabus, any lecture materials they generated for the mini‐course, and peer and faculty evaluations from their teaching. Mid‐semester student feedback was positive with one exception. The primary criticism given by the students was that too much in‐class time was dedicated for working on the mini‐course and that students desired more time to be dedicated to delivering content. This was adjusted mid‐course and in‐class time for working on the mini‐course was eliminated for the final four weeks of the instructional block. Based on student feedback, this dedicated in‐class time was no longer useful after week three and future versions of this course will reflect that. Overall, students reported through evaluations that the course was beneficial and significantly improved their confidence in conducting lectures, engaging in active learning techniques, and in preparing for future teaching. Several students reported being able to immediately apply the knowledge and techniques in other courses in which they teach directly or provide supplemental instruction. Course directors and faculty from those courses noted the improved performance of those students in those duties. In conclusion, we report that student teaching performances and their use of active learning is improved after participation in this new course. In addition, students self‐report that the course has increased their confidence in conducting science teaching. The addition of learning best‐practices in teaching may be important for optimizing trainee success.

Support or Funding Information

Supported by the American Physiological Society and the National Science Foundation IUSE: Education and Human Resources (DUE 1822245).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
PhUn Week: Bigger and Better
Authors: Rascon Jozelyn , Trujillo Elizabeth , Pena Marcelo , Pena Francisco J , Ramirez Juliana A , Melendez Gladys , Lazcano Cameron , Gurovich Alvaro N
Published: June 2021

Physiology Understanding (PhUn) week is the American Physiological Society's K‐12 outreach program. In general, PhUn week consists of hands‐on activities and presentations, which incorporate the understanding of physiology through interactive fun. In 2017, our PhUn week activity reached 24 seventh‐grade students and they performed three activities using electronic physiology data acquisition systems (Biopac MP40). Even though previous PhUn week events were already structured, we wanted to reach more school‐grade students. Therefore, we collaborated with Tippin Elementary School in El Paso, Texas and its whole second grade (96 students) and teachers (5). Because of the larger demand, we designed 6 different activities to be performed at PhUn week: Reaction time, changes in heart rate with exercise, and electromyography activity with different exercises (Biopac MP40, previously performed) and three new activities (i.e. respiratory system, sensory system, and cardiovascular system). In addition, we recruited 14 more undergraduate students, from the Bachelors in Rehabilitation Sciences at UTEP, and trained them in the use of Biopac MP40 and the new activities. The UTEP undergraduate students that participated in PhUn week the previous year performed this training. The day of the presentation, the multipurpose room of the school was converted into a physiology activity room with six different stations (2 of each) for a total of twelve set up for the students to rotate between. The stations were the reaction time station, electromyography station, electrocardiogram station, a station where students built a working model of the diaphragm and lungs, a sensory challenge station where students searched in a bin of rice for miscellaneous objects, and finally an interactive working model of the heart. We were able to do 15‐minute rotations so all 96 second grade students were able to visit each station. The students, teachers, and other Tippin faculty, as well as UTEP students had a lot of fun and learned about physiology at the same time. We could see the excitement and effect the event left on the young students as many left excited about science, which is the main goal of PhUn week. Overall, it was a great experience that required a lot of preparation and training but was worth the impact it had on the students involved.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Using Biotensegrity to Explain the Mechanically‐Induced Lesions in Navicular Disease
Authors: Osborn Michelle L , Blas‐Machado Uriel , Kirejczyk Shannon GM , Uhl Elizabeth W
Published: June 2021

Degenerative joint disease (DJD), the most common disease affecting humans and animals, has long been believed to result from failure of the joints from ‘wear and tear’ in which damage exceeds repair. However, models that depict the body as a machine do not adequately explain the functional musculoskeletal anatomy, in particular the role of fascia, nor do they explain why the lesions of DJD do not always correlate with pain and dysfunction. In contrast a body structured through tensegrity (integrity by tension; biotensegrity) has a seamless integration of structure and function from the molecular, cellular, tissue and organ levels to the whole body, and better explains how life evolved by adapting to different environments. Joints structured through tensegrity have more functional flexibility as tensegrity allows mechanical forces to be transmitted and dispersed throughout the surrounding connective tissues and counteracted by muscular contractions rather than being transmitted as compression across a joint space, which is the case in machine models. The purpose of this study was to use biotensegrity to explain the pathogenesis of tissue damage in navicular disease, a devastating form of DJD in horses. The structure of the navicular apparatus indicates it is an enthesis organ that functions through tensegrity to dissipate forces away from the osteotendinous junction between the deep digital flexor tendon and the distal phalanx. Functional analyses of 3D models reconstructed from CT scans of a horse's forelimb were used to predict where mechanical forces were most impacting the navicular enthesis organ. These models show that pronounced dorsiflexion of the foot, a posture commonly observed in horses with navicular disease, increased the mechanical forces on the deep digital flexor tendon, navicular bone, impar and collateral sesamoidean ligaments, which are all sites of tissue damage in navicular disease. Histochemical staining with Masson's trichrome and picrosirius red, confirmed evidence of mechanical stress in the target tissues as stretched collagen was prominent in the tendon, navicular bone and fibrocartilage, and the impar and collateral sesamoidean ligaments and correlated with the sites of tissue damage. The identification of widespread mechanical damage in the soft tissues of the navicular enthesis organ in affected horses confirms its function as a tensegrity structure and emphasizes the relevance of biotensegrity models in understanding the pathogenesis of DJD.

Support or Funding Information

LSU SVM Charles V. Cusimano Equine Health Studies Program

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Groove for the Sphenoparietal Sinus: A Constant Cranial Structure in Human Gross Anatomy
Authors: Parra Jorge Eduardo Duque , Ríos John Eduardo Barco , García Juan Fernando Velez
Published: June 2021

Objectives

Cranial venous sinuses are elements formed by dura mater, they are fixed in bony parts leaving their tracks as grooves, which can be used as recognition in morphological studies. One of these grooves, the sphenoparietal sinus groove, is not referenced in Terminologia Anatomica, so the objective is to demonstrate the existence in human skulls.

Methods

Sixty five human adult skulls were selected, without distinction of ethnic group or sex, in order to evaluate the existence of a bony groove for the sphenoparietal sinus. As inclusion criteria, the skulls had to have integral the anterior region of the middle cranial fossa. To establish the existence of the groove, under the lesser wing of the sphenoid was visualized and palpated endocranially.

Results

Of the 65 skulls, 63 had a bony groove related to the sphenoparietal sinus (96,92%); of these, 50 presented bilaterally (76,92%) and 13 unilaterally: 9 in the left region (13,84%) and 4 in the right (6,15%); two skull did not present the groove (3.08%).

Conclusion

In Terminologia Anatomica, the groove for the sphenoparietal sinus is not described, a constant structure due to its high percentage of presence (96,92%), for which the term “groove for the sphenoparietal sinus” is proposed, which can serve as a landmark in cranial morphology.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Guillemeau: “Les oeuvres chirurgicales” (surgical works), Rouen (France) 1649
Authors: LE FLOCH‐PRIGENT Patrice Pierre , NGUYEN Juliette , GILLOT Jean Bernard
Published: June 2021

Jacques GUILLEMEAU (1550–1612) published several books during his surgical life. He was a brilliant follower of Ambroise Paré and the surgeon of several kings of France. This late edition was a posthumous and complete one, published in the town of Rouen in 1649 by Jean VIRET and printed by Pierre MAILLE. It was an in folio with 863 pages and numerous figures: 32 copper plates = 21 anatomical ones and 11 plates of surgical instruments. There were also 18 wooden plates with foetus in utero and paediatric instruments. The exact title was “ Les oeuvres chirurgicales. Avec les portraicts et figures de toutes les parties du corps humain, & des instruments nécessaires au chirurgien...”. We studied particularly the anatomical part. The obstetrical and paediatrical parts were also famous.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Activity of LHX2‐associated cis‐Regulatory Modules During Limb Development
Authors: Roberts Kari D , Mulder Cassidy G , Shah Neera M , Watson Billy A , Pira Charmaine U , Shin Alexandra , Oberg Kerby C
Published: June 2021

Limb development occurs along three major axes – proximal‐distal, anterior‐posterior, and dorsal‐ventral. Fibroblast growth factors (FGFs) secreted by the apical ectodermal ridge (AER) and sonic hedgehog (SHH) released from the zone of polarizing activity (ZPA) are responsible for proximal‐distal and anterior‐posterior development, respectively, and maintain each other through a positive feedback loop. This reciprocal loop is critical for proper limb development. Recently, we identified LIM homeobox 2 (LHX2) as an intermediate in FGF‐mediated SHH expression. There are over 25 conserved regions of non‐coding DNA associated with the LHX2 gene locus that could serve as regulatory modules and targets of FGF signaling. We hypothesize that FGF regulates LHX2 through at least one of these potential cis regulatory modules (PCRMs). We selected 10 PCRMs with active chromatin marks in the limb and screened for their activity within the LHX2 expression domain (distal mesoderm subjacent to the AER). Each PCRM was inserted into the pTK‐GFP reporter and electroporated into the distal mesoderm of HH20‐23 chicken embryo wing buds. PCRM activity was determined 24 hours later using fluorescence microscopy. We found three of the PCRMs display activity (CRM (−19), CRM (−2), and CRM (−1)) that overlap LHX2 expression in the chicken wing bud. One CRM (−19), approximately 130 kb upstream of the LHX2 locus, is most consistent with the pattern of LHX2. Further studies are underway to determine whether these CRMs interact with the LHX2 promoter and whether FGF regulates their activity.

Support or Funding Information

Funded in part by a grant from the LLU Pathology Endowment.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
MicroRNA‐29b act as mediators of lipopolysaccharide‐induced bone loss
Authors: Sul OJ , Choi Hye‐Seon
Published: June 2021

MicroRNAs (miR) play critical roles in osteoclastogenesis, resulting in regulating bone density. Since the expression level of miR‐29b was increased in tibiae from lipopolysaccharide(LPS)‐injected mice and LPS‐induced osteoclasts (OCs). we hypothesized that LPS induces miR‐29b, which enhance OC formation, leading to excessive bone loss. Overexpression of miR‐29b increased OC survival without any change in OC differentiation, whereas inhibition of endogenous miR‐29b induced by LPS decreased it. Increased OC survival upon overexpression of miR‐29b was due to anti‐apoptotic activity evaluated by staining annexin V‐positive cells. We found that a target gene of miR‐29b is Bcl2 modifying factor (Bmf), which acts as a proapoptotic factor, and that miR‐29b binds to the 3′‐UTR of Bmf. Our data demonstrate that LPS‐induced miR‐29b increases the number of OC by enhancing OC survival through decreased BMF.

Support or Funding Information

This work was supported by the Basic Science Research Programs (2018R1A2B6001276;2016R1A6A3A11932375 2014R1A6A1030318) of the NRF and Korea Healthcare Technology R&D Project (HI18C0375) funded by the Ministry of Health.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Endogenous siRNAs are required for a learned pathogen avoidance behavior in Caenorhabditis elegans
Authors: Manzanet Yazmine , Salerno Nicholas , Youngman Elaine M
Published: June 2021

The nematode C. elegansis a premier model for the study of learning and exhibits a variety behavioral plasticity phenotypes. For example, worms that are infected by the pathogen Pseudomonas aeruginosa (PA14) early in life associate pathogen odors with infection, and learn to avoid the pathogen as adults when given a choice between PA14 and a nonpathogenic E. coli food source (OP50). This associative learning requires an increase in serotonin in the ADF chemosensory neurons, but the remaining molecular circuitry that allows learned pathogen avoidance is unknown. Based on the known role of small noncoding RNA molecules in regulating other behavioral responses in the worm, we have used a genetic approach to determine the involvement of endogenous small RNAs in pathogen avoidance behavior. Mutants that are defective in small RNA pathways were raised in an environment with both pathogenic PA14 and nonpathogenic OP50, and their pathogen avoidance behavior as adults was compared to that of wild‐type worms using a standard choice assay. For each genotype, we have completed at least three replicate choice assays of at least 71 worms each for the naïve and trained conditions. Briefly, the choice assay involves placing either trained or naïve worms in the center of a standard Nematode Growth Medium plate, and worms are allowed to choose between PA14 and OP50 food sources spotted on opposite sides of the plate. The number of worms found in the PA14 and OP50 spots is counted after an hour, and the proportion of worms on PA14 is compared between naïve and trained worms using a one‐tailed t‐test. As previously published, trained wild‐type worms (raised in the presence of pathogen) show increased avoidance of PA14 when compared with naïve worms (p=0.04369). Interestingly, mut‐7(pk204)animals, which lack a large class of small RNAs known as the endogenous siRNAs, show no change in pathogen avoidance when raised under naïve or trained conditions (p =0.86258), suggesting that endogenous siRNAs are required for learned pathogen avoidance. mut‐7is an upstream component of several functionally and genetically distinct endogenous siRNA pathways, including the 26G‐RNA and 21U RNA‐triggered pathways, the CSR‐1 pathway, and the WAGO pathway. To determine which of these pathways is specifically required for learned pathogen avoidance behavior, we are currently testing learning in mutants that disrupt individual pathways and will report on these results.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.