bluuebunny/crossref_metadata_2025 huggingface dataset:
Administration of a selective prostaglandin E2 receptor agonist attenuates renal fibrosis in vitro and in vivo
Authors: Jensen Michael Schou , Mutsaers Henricus A.M. , Tingskov Stine Julie , Christensen Michael , Madsen Mia Gebauer , Olinga Peter , Kwon Tae‐Hwan , Norregaard Rikke
Published: June 2021

Renal fibrosis plays a pivotal role in the development and progression of chronic kidney disease (CKD), which affects 10% of the adult population. Previously, it has been demonstrated that the cyclooxygenase‐2 (COX‐2)/prostaglandin (PG) system influences the progression of renal injury. PGE2 interacts with four different G‐protein‐coupled PGE2 EP receptors, known as EP1–EP4, and plays a role in several biological functions. Previous studies show that the EP2 receptor is involved in progression of lung fibrosis. In this study, we investigated the effect of an EP2 agonist (butaprost) on renal fibrogenesis on a cellular, renal tissue and organismal level using well‐established in vivo and in vitro models as well as a recently developed human model of renal fibrosis. This model is ideal to study multicellular pathological processes, e.g. fibrosis, directly in human tissue since cellular diversity and organ architecture is maintained in the slices. Butaprost (50 μM) reduced TGF‐β‐induced fibronectin (FN) expression, Smad2/3 phosphorylation and epithelial‐mesenchymal transition in MDCK cells. In addition, treatment with 4 mg/kg butaprost attenuated the development of fibrosis in mice that underwent unilateral ureteral obstruction (UUO) surgery, as illustrated by a significant reduction in the gene and protein expression of α‐smooth muscle actin, FN and collagen 1A1. More importantly, a similar anti‐fibrotic effect of butaprost was observed in human precision‐cut kidney slices exposed to TGF‐β. The mechanism of action of butaprost appeared to be a direct effect on TGF‐β/Smad signaling, which was independent of the cAMP/PKA pathway. In conclusion, this study demonstrates that stimulation of the EP2 receptor effectively mitigates renal fibrogenesis in various fibrosis models. These findings warrant further research into the clinical application of butaprost, or other EP2 agonists, for the inhibition of renal fibrosis.

Support or Funding Information Sources of support: This work was kindly supported by Lundbeckfonden, grant number R231‐2016‐2344 as well as the Danish Council for Independent Research, grant number 6110‐00231B, Aarhus University Research Foundation, grant number AUFF‐E‐2015‐FLS‐8‐69 and Hildur and Dagny Jacobsens Foundation grant number 1295716‐1.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Reno‐protective Effect of Central Leptin Receptor Blockade in Rat Model of Menopause
Authors: Van Erdewyk Jonathan , Khan Bilal , Babich Victor , Barnes Maria , Di Sole Francesca
Published: June 2021

Women in menopause have substantial variations in hormonal balance. The weight gain in the form of adipose tissue in these women has been associated to an increase in leptin level; an adipocyte‐derived hormone that acts primarily on the central nervous system. The increase in central leptin has been related with the development of hypertension via an increase in renal sympathetic nerve activity (RSNA) and activation of RSNA has been demonstrated to play a role in the progression of kidney dysfunction. Therefore, we hypothesize that activation of RSNA due to elevated leptin induces hypertension via an action on kidney function. Utilizing a menopausal rat model, we studied the mechanisms mediated by central leptin receptor activation to alter kidney function. We measured the glomerular filtration rate (GFR) by creatinine clearance and protein level of the neutrophil gelatinase‐associated lipocalin (NGAL), an early biomarker of kidney injury, by immunohistochemistry. An increase in creatinine clearance (from 1.3 ml/min to 2.3 ml/min) and protein levels of NGAL (over 90% increase in both glomeruli and tubules) was measured in ovariectomized adult female rats when compared with their sham‐operated controls. Treatment with leptin reception antagonist (LAN‐6, 3μg/day), delivered for four weeks into the lateral ventricle by an osmotic pump, significantly attenuated these effects. Furthermore, we determined that intra‐abdominal and perirenal fat accumulation were both markedly higher in the same menopausal rat models. Perirenal fat accumulation has been associated with lipid accumulation in renal tissues and excess of intrarenal lipid is increasingly recognized to contribute to kidney injury through lipotoxicity. We found a significant accumulation of renal lipid, tested using oil red O staining, in the renal tubules of the ovariectomized animals when compared with their controls. The lipid accumulation was partially reversed by the leptin receptor antagonist treatment. In summary, the increase in GFR associated with kidney injury suggests the development of kidney disease in this rat model of menopause that is in part reversed by blocking central leptin receptors; infiltration of lipid in the renal tissue is proposed as a possible mechanism of kidney injury. These findings have the potential to translate into future therapeutic strategies for treating kidney injury in menopausal mediated‐hypertension.

Support or Funding Information

Iowa Osteopathic Education and Research Funds

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Salt‐Responsive Metabolite, beta‐Hydroxybutyrate, Attenuates Hypertension
Authors: Chakraborty Saroj , Galla Sarah , Cheng Xi , Yeo Ji‐Youn , Mell Blair , Singh Vishal , Yeoh BengSan , Saha Piu , Mathew Anna V. , Vijay‐Kumar Matam , Joe Bina
Published: June 2021

Dietary salt reduction and exercise are lifestyle modifications for salt‐sensitive hypertensives. While exercise has prominent metabolic effects, salt has an adverse effect on metabolic syndrome, of which hypertension is a hallmark. We hypothesized that dietary salt impacts metabolism in a salt‐sensitive model of hypertension. An untargeted metabolomics approach demonstrates lower circulating levels of the ketone body, beta‐hydroxybutyrate (βOHB), in high salt‐fed hypertensive rats. Despite the high salt intake, specific rescue of βOHB levels by nutritional supplementation of its precursor, 1,3‐butanediol, attenuates hypertension and protects kidney function. This beneficial effect of βOHB was likely independent of gut‐microbiotal and Th17‐mediated effects of salt and instead facilitated by βOHB inhibiting the renal Nlrp3 inflammasome. The juxtaposed effects of dietary salt and exercise on salt‐sensitive hypertension, which decrease and increase βOHB respectively, indicate that nutritional supplementation of a precursor of βOHB provides a similar benefit to salt‐sensitive hypertension as exercise.

Support or Funding Information

Funding from this work to BJ from the NHLBI (NIH) (HL143082) is gratefully acknowledged.

Declaration of interests: Bina Joe and Saroj Chakraborty have an interest in U.S. Patent Application Serial No.: 62/665,690 filed on May 2, 2018 in this paper regarding nutritional intervention for salt‐sensitive hypertension for 1,3‐butanediol.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
High Sodium Diet in 11β Hydroxysteroid Dehydrogenase Type 2 CNS Knockout Mice Induces a Proinflammatory Phenotype of Perivascular Adipose Tissue and Alterations in Arterial Reactivity
Authors: Grenier Celine , Ralph Ailsa , Livingstone Dawn EW , Holmes Megan C , Bailey Matthew A
Published: June 2021

Rationale

We previously generated CNS‐specific knockout of HSD2 (“brain” knockout; BKO), showing that the mice have inappropriate salt‐appetite and hypertension under a high sodium diet (Evans, Ivy et al. 2016) . Multiple peripheral components may contribute to this salt‐sensitivity and here we have focussed on the role of resistance arteries, which are surrounded by perivascular adipose tissue (PVAT), able to secrete vasoactive molecules (pro‐ or anticontractile). It is well known that PVAT interacts with arteries to regulate their function. Studies also show that high‐sodium diet stimulates adipocity and alters adipocyte function. Our aim is to study the impact of high sodium intake on PVAT gene expression and on arterial reactivity in BKO knockout mice.

Methods

BKO mice were generated by crossing HSD2flx/flx with Nestin‐Cre mice; HSD2flx/flx littermates were used as wild‐type controls (WT). Mice were fed with high sodium diet (HSD; 3% Na) or normal sodium diet (NSD; 0.3% Na) for one week before being culled. The PVAT and the mesenteric arteries were dissected. The arterial reactivity to phenylephrine (Phe), acetyl choline (Ach) and sodium nitroprusside (SNP) was studied by wire myography. In PVAT from mesenteric arteries, the expression of a panel of pro or anti inflammatory markers was measured by qRT‐PCR.

Results

On normal sodium diet, There was no difference between genotypes in the PVAT expression of a panel of genes reporting inflammation (adipoq, lep, cybb, ptgs2, agtr1a, serpin, pparg, il‐6, ccl2, adgre1, vcam1, atp6ap2, nampt). High sodium diet demonstrated a significant (all P<0.05) overexpression of the proinflammatory genes il‐6, ccl2, cybb and ptgs2 on BKO fed with HSD compared to WT; other genes were not different. In PVAT‐free mesenteric arteries, hsd11b2 gene expression was not different between WT and BKO mesenteric arteries and on normal sodium diet, arterial reactivity to Phe, Ach and SNP was not different between genotypes. In contrast, 7 days of high sodium intake revealed there were differences in arterial function between genotypes: there was a significant increase in sensitivity to Phe in the WT and a decrease of the maximum effect of SNP in the BKO mice.

Conclusion

The high salt diet on BKO mice led to an overexpression in pro‐inflammatory gene in the PVAT surrounding mesenteric arteries and affected intrinsic arterial reactivity. The interaction between these two phenomena remains to be elucidated but it may contribute explanation to the increase of blood pressure of salt sensitive BKO mice.

Support or Funding Information

This work was funded by The British Heart Foundation (PG/16/98/32568)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Activation of G Protein‐Coupled Estrogen Receptor 1, with a GPER Agonist, Reduces Blood Pressure in Female Growth‐Restricted Rats in Later Life
Authors: Davis Gwendolyn K. , Bamrick Daniel R. , Newsome Ashley D. , Coats Laura E. , Alexander Barbara T.
Published: June 2021

The prevalence of hypertension (HTN) is two‐fold higher in low birth weight (LBW) women at age 60 relative to their normal birth weight (NBW) counterparts. Yet, the mechanisms that contribute to greater cardiovascular risk in LBW women are not known. In our model of intrauterine growth restriction (IUGR), induced by placental insufficiency in the pregnant rat, female IUGR offspring have an elevated blood pressure (BP) at 12 months of age that is associated with early reproductive senescence. Although estrogen levels do not differ between control and IUGR groups, there is a significant increase in testosterone levels in female IUGR that is not seen in female control. Elevated testosterone levels, due to an imbalance in the hormonal milieu, is suggested to contribute to the development of hypertension in women in later life. We previously reported that chronic administration of estradiol is able to abolish the increase in blood pressure in the female IUGR rat at 12 months of age. However, this was not accomplished through alterations in expression of the nuclear estrogen receptor alpha (ER‐alpha). In the current study, we tested the hypothesis that activation of G protein‐coupled estrogen receptor 1 (GPER) would reduce blood pressure in the female IUGR rat. The selective GPER agonist, G1 (400 g/kg/day), or vehicle was administered via osmotic pumps for 2 weeks beginning at 14 months of age. BP was measured in conscious, chronically catheterized rats following treatment (2way ANOVA; Tukey's multiple comparison). As shown previously, BP was significantly elevated in vehicle‐treated IUGR relative to vehicle‐treated control (138.3± 4.01 vs. 124.3 ± 0.70mmHG, respectively). BP was significantly decreased in G1‐treated IUGR relative to vehicle‐treated IUGR (125.14±4.03 vs. 138.3 ± 4.01mmHG, respectively), whereas BP was not significantly altered in G1‐treated control relative to vehicle‐treated control (124.5 ± 0.66 vs. 124.3 ± 0.70mmHG, respectively). Uterine weight did not differ between groups. Therefore, this data suggests that activation of GPER was able to abolish the increase in blood pressure in the female IUGR rat in later life. This study suggests that GPER may serve as a therapeutic target for ameliorating increases in blood pressure seen in LBW women in later life.

Support or Funding Information

NIH: R56HL143459, P20GM121334, P20GM104357, P01‐HL51971, T32HL105324, F30DK112718. AHA: GRNT19900004, PRE34060010.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
AT1a stimulation of tonicity‐responsive enhancer binding protein (TonEBP/NFAT5) translation through Annexin‐A2 may represent allostatic anticipation of increased tonicity
Authors: Hultström Michael , Paliege Alexander , Jönsson Sofia , Agic Mediha Becirovic , Melville Jacqueline , Skogstrand Trude , Fähling Michael
Published: June 2021

Allostasis is idea that the body may anticipate homeostatic challenges and regulate cell and organ function in preparation. Angiotensin II (ANGII) is important for regulating sodium balance and as such increases tonicity in the renal medulla. The present study identifies a novel regulatory pathway whereby ANGII increases the translation of tonicity‐responsive enhancer binding protein (TonEBP/NFAT5) by activating nucleic acid binding of Annexin‐A2, which has several binding sites on NFAT5 mRNA and increases its translation. The activation of Annexin‐A2 was shown using heparin affinity chromatography and mass spectrometry in four‐week two‐kidney, one‐clip hypertension, and verified in two‐week ANGII infusion in rats. The addition of the AT1‐receptor blocker Losartan reduced the activation. Renal gene expression was studied using micro arrays, and identified NFAT5 targets as enriched. Western blot showed that NFAT5 protein was increased, while mRNA was not significantly regulated consistent with posttranscriptional regulation by Annexin‐A2. In conclusion, ANGII/AT1/Annexin‐A2‐mediated increase of the normally tonicity‐regulated NFAT5 in the kidney may be a way to ensure a more robust response to ANGII‐induced tonicity increase, and thus better protect homeostasis.

Support or Funding Information

Lars Hierta Foundation, Magnus Bergvall Foundation, Swedish Society for Medical Research.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Microbiota and Xenobiotics Reveal Links between Myopathy, Inflammaging and Glycolytic Metabolism
Authors: Schertzer Jonathan
Published: June 2021

Environmental cues influence the risk and severity of myopathy and sarcopenia. Increased inflammation during ageing (i.e. inflammaging) has been proposed as a contributor sarcopenia. The source of age‐related inflammation is ill‐defined. Xenobiotics, including therapeutic drugs, can also promote inflammation that contributes to myopathy. However, the participatory immune components that link either of these stressors to functional decline in skeletal muscle are unknown. The intestinal microbiota has emerged as an environmental factor that can alter xenobiotic metabolism, inflammation and host metabolism. We sought to define aspects of the microbiota and specific innate immune components that connect triggers of inflammation during ageing or xenobiotic exposure.

We used statins as a xenobiotic given well documented statin myopathy. Despite the widespread anti‐inflammatory effects, statins actually increase a very specific immune response that is propagated by an inflammasome that acts as a metabolic danger sensor. Statins activate the NOD‐like receptor family, pryin domain containing 3 (NLRP3) inflammasome. In order to implicate the microbiota, we used aged germ free mice and mice lacking bacterial cell wall sensors to test how the entire microbiota alters sarcopenia.

We hypothesized that NLRP3‐null mice would be protected from statin‐induced myopathy and sarcopenia compared to wild type (WT) mice. We hypothesized that germ free mice (devoid of any bacteria) and Nod1‐null would be partially protected from age‐related inflammation and sarcopenia.

We found that NLRP3‐null mice had an attenuation of statin myopathy and sarcopenic indicators compared to WT mice. A common mechanism between sarcopenia and statin myopathy was lower glycolysis in muscle cells. Further, specific bacterial ligands such as lipopolysaccharide exposure magnified statin myopathy. However, we found that germ free mice had worse indicators of sarcopenia compared to conventional mice. We found that Nod1‐null mice had worse indicators of sarcopenia compared to WT mice.

Our results show that xenobiotics and aging alter glycolytic metabolism, which is a common mechanism linking inflammasome responses to myopathy from various stressors. Specific bacterial components play a role in augmenting statin‐induced myopathy via the NLRP3 inflammasome. However, the totality of the bacterial community and bacterial cell wall sensing actually protects against sarcopenia.

Support or Funding Information

Funded by Natural Sciences and Engineering Research Council of Canada (NSERC) and the Canadian Institutes of Health Research (CIHR).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Comparison of the Effects of Cryotherapy and Thermotherapy on Permeability After Skeletal Muscle Injury
Authors: Gasser Brittany , Tusavitz Sabrina , Carpenter April
Published: June 2021

Skeletal muscle injuries are common in active individuals. The inflammatory process is a vital component of muscle healing beginning with enhanced permeability. As blood vessels dilate, inflammatory promoting proteins and white blood cells enter the damaged area to eliminate debris and initiate regeneration. The application of controlled temperature on the injured muscle remains the most common treatment. However, there is conflicting evidence on the advantages of cryotherapy, and there is a lack of evidence on benefits of thermotherapy. Therefore the purpose of our study was to compare the effects of cryotherapy and thermotherapy on muscle vascular permeability in both male and female mice. Barium chloride (BaCl2) was injected into the right tibialis anterior (TA) to cause a chemical injury and sodium chloride (NaCl) was injected into the left TA as a control. Therapy was started immediately after injury. The control group was anesthetized, the thermotherapy group received heat treatment (41°C) while anesthetized, and the cryotherapy group received cold therapy (10°C) while anesthetized, for 20 mins every 3 hours, during 2 consecutive 12 hr wake cycles. Evans Blue (EB) dye was injected through the lateral tail vein to assess vascular permeability in the TA muscles. There was no significant difference in permeability between the NaCl injected muscles regardless of treatment. Permeability 36 hours post‐injection was significantly elevated in the BaCl2 injected TA muscles in all groups. Cryotherapy significantly reduced permeability in the BaCl2 injured muscles as compared to thermotherapy (cryotherapy: 4.78±2.68 vs. thermotherapy: 11.1±2.32 ngEB/mgTA). Our data suggests that cryotherapy limits permeability 36 hrs post‐skeletal muscle injury. New data suggests inflammation is necessary for efficient muscle healing. Therefore new treatments for muscle injury should be considered.

Support or Funding Information

Ursinus College Student Research Funding

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
MG29 Interacts with Bin‐1 to Maintain T‐Tubule Structure in Skeletal Muscle Physiology and Regeneration
Authors: Yi Frank , Zhou Xinyu , Gumpper Kristyn , Zhu Hua
Published: June 2021

Mitsugumin 29 (MG29), a member of the synaptophysin‐like family proteins, is a transmembrane protein primarily expressed in the t‐tubule membranes of skeletal muscle. In the current study we have found that t‐tubules in skeletal muscle of mg29−/− have defective morphology, indicating importance of MG29 in maintenance of muscle structure and function. To elucidate the role of MG29 in muscle physiology, we performed two‐color STORM super‐resolution imaging analysis confirmed co‐localization of MG29 and Bin‐1 on t‐tubules of skeletal muscle. We also performed co‐immunoprecipitation and found that MG29 can bind to Bin‐1, another t‐tubular protein. Recombinant MG29 protein domain fragments tagged with GST were expressed and Bin‐1 was found to bind to the cytosolic domains of MG29. Furthermore, organized distribution of Bin‐1 is severely disrupted in mg29−/− muscle. To test the function of MG29 in muscle regeneration, we injured gastrocnemius muscle with cardiotoxin (CTX) and tracked muscle repair and regeneration. Western blot showed that following CTX injury, MG29 protein levels were transiently reduced from day 1 to day 3, followed by recovery associated with muscle regeneration by day 5. Protein level of Bin‐1 in wild type muscle was also reduced upon CTX‐injury and increased during the recovery process. Compared with wild type muscle, the mg29−/− muscle displayed delayed regeneration and increased fibrosis following CTX‐induced injury. Together our data suggest that functional interaction between MG29 and Bin‐1 contribute to maintenance and formation of t‐tubule network while playing a role in muscle regeneration and regeneration. Targeting this MG29‐Bin‐1 interaction might provide a potential effective approach for treatment of muscle diseases

Support or Funding Information

This work was supported by grants from the National Institutes of Health (NIH) (AR067766 and HL124122)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Whole Body Hyperthermia Ameliorates Inflammation via Regulating Pro‐inflammatory but not Anti‐inflammatory Macrophages in Tenotomized Rat Skeletal Muscle
Authors: Hirunsai Muthita , Srikuea Ratchakrit
Published: June 2021

Following disuse condition, macrophages play significant role in inflammatory responses during muscle degeneration and repairing processes. Pro‐inflammatory (M1 phenotype) macrophage becomes classically activated during an initial phase of muscle degeneration. Contrariwise, anti‐inflammatory (M2 phenotype) macrophage abrogates inflammation by deactivating M1 macrophage and promotes muscle growth in repairing stage. Previously, we demonstrated that heat treatment alleviates the loss of muscle mass via enhancing intramuscular capillarization and remodeling extracellular matrix in disuse muscle. Of these, the effect of heat stress on inflammatory responses in tenotomy‐induced atrophy is currently unknown. In this study, the effect of hyperthermia on M1 and M2 macrophage populations as well as the responses of inflammatory cytokine protein expression in the soleus muscle following Achilles tendon transection (tenotomy) were examined. Male Wistar rats were designed into sedentary control (CON), tenotomy without heat treatment (TEN), and tenotomy plus heat treatment (TEN+HEAT) groups. Whole body hyperthermia was maintained in TEN+HEAT group with animal core body temperature at 40.5–41.5°C, 30 min for 7 days. After 8 days of tenotomy, soleus muscle demonstrated a reduction in fiber size and tremendous infiltration of inflammatory macrophages. These occurrences coincided with an increase in pro‐inflammatory cytokine, particularly tumor necrosis factor‐alpha (TNF‐α). Whilst tenotomy had no effect on interleukin‐10 (IL‐10) which is an anti‐inflammatory cytokine. Interestingly, whole body hyperthermia diminished accumulation of M1 macrophages and associated with reduction of TNF‐α protein expression in tenotomized muscle. Nevertheless, heat stress affected neither M2 macrophage phenotype invasion nor IL‐10 protein level following tenotomy. Collectively, heat stress relieved tenotomy‐induced inflammatory response, partly by inhibiting M1 macrophage invasion. Our findings provide a foundation for therapeutic strategy of heat stress on fiber inflammation during skeletal muscle atrophy.

Support or Funding Information

The research was supported by the grant from Srinakharinwirot University (to MH).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Renal Dysfunction Exacerbates Ischemic Muscle Injury in Mice Subjected to Hindlimb Ischemia
Authors: Berru Fabian N. , Gray Sarah E. , Thome Trace , Le Dennis , Omalley Kerri A. , Berceli Scott A. , Scali Salvatore T. , Ryan Terence E.
Published: June 2021

Patients with critical limb ischemia (CLI) have multiple concomitant comorbidities which are associated with poorer overall health outcomes. CLI patients with impaired kidney function and/or diabetes are at greater risk for limb amputation and death. While the negative impacts of renal dysfunction and diabetes on CLI outcomes are known clinically, the underlying biologic mechanism(s) responsible are poorly understood. Thus, we sought to evaluate the impact of impaired renal function and diabetes on the skeletal muscle pathology in mice following unilateral hindlimb ischemia (HLI). Renal dysfunction (RD) was induced by dietary administration of adenine which causes renal tubular injury, while diabetes was induced by streptozocin injection. HLI was induced by ligation and excision of the femoral artery, while the contralateral hindlimb was utilized as a control (no surgery). Three groups (n = 15; 5 per group) of C57BL/6J mice were analyzed: control, RD, and RD+ diabetes. Skeletal muscle ischemic injury was assessed using histological measurements of myofiber centralized nuclei, myofiber cross‐sectional area, ischemic lesion areas, and vascular density. High‐resolution O2 consumption was used to examine mitochondrial function in permeablized myofibers. Mice with RD, regardless of diabetes, exhibited greater ischemic lesions within the tibialis anterior muscle (Con = 2.53 ± 0.51% vs. RD = 60.554 ± 24.15% vs. RD+STZ = 79.99 ± 19.99%; P < 0.0001) despite similar capillary density (Con = 466.36 ± 44.98 vs. RD = 455.20 ± 79.55 vs. RD+STZ = 578.20 ± 87.16 capillaries/field; P = 0.9011). Mice with RD exhibited a ~45% decrease in mitochondrial respiratory capacity across numerous substrates, whereas control mice had normal respiration in the ischemic limb muscle seven days post‐HLI. These observations demonstrate that impaired kidney function exacerbates ischemic muscle injury in mice. Future studies are needed to uncover the molecular mechanisms by which RD negatively impacts skeletal muscle.

Support or Funding Information

Supported by AHA Grant 18CDA34110044 to TER.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Functional Role of microRNAs in Patient‐Derived Xenograft Models of Human Cholangiocarcinoma
Authors: Wu Nan , McDaniel Kelly , Glaser Shannon , Celinski Scott , Dhruv Harshil , Alpini Gianfranco , Meng Fanyin
Published: June 2021

BACKGROUND & AIMS

Patient‐derived xenograft (PDX) models are now being widely used in cancer research including liver and biliary carcinogenesis, and have the potential to feedback and improve the therapeutic effects in cancer patients. MicroRNAs (miRNAs) are highly conserved 19–25 nucleotide non‐coding small RNAs which are primarily involved in gene silencing. Our aim was to characterize the functional roles of microRNA regulated protein translocations in patient‐derived cholangiocarcinoma xenograft tumors.

METHODS

miRNA expression in human cholangiocarcinoma cancer tissues, patient‐derived cholangiocarcinoma xenograft tissues, human extrahepatic cholangiocarcinoma cancer stem cells (GB‐CSCs)/Mz‐ChA‐1 extrahepatic cholangiocarcinoma cancer cells and their derived xenograft tumor tissues, as well as H69 non‐malignant cholangiocytes was assessed using a hybridization based microarray and was further evaluated by Taqman real‐time PCR analysis. Promoter methylation, matrix metalloproteinase (MMP) 1, 2, 3 and 9 mRNA expression was quantitated by real‐time PCR and western blot analysis.

RESULTS

We identified miR‐200 family including miR‐200a, miR‐200b and miR‐200c that are differentially expressed in doxorubicin‐enriched cell fractions in a patient derived mouse xenograft tumor model of extrahepatic cholangiocarcinoma. Members of the miR‐200 family were notably more silenced in GB‐CSCs compared with their parental extrahepatic cholangiocarcinoma cancer cells (GBCs) and Mz‐ChA‐1 cells. Expression of miR‐200 family microRNAs and transmembrane protein MMP‐2 and MMP‐9 were substantially enhanced concomitant with GBCs differentiation and loss of self‐renewal, whereas forced expression of a polycistron construct encoding miR‐200 miRNAs significantly decreased chemoresistance to doxorubicin. Methylation‐specific PCR detected that miR‐200b and miR‐200c promoter was hypermethylated in GB‐CSCs, GBCs as well as Mz‐ChA‐1 cells. Cell migration and invasion were significantly decreased by miR‐200 precursor introduction in GB‐CSCs and Mz‐ChA‐1 cells. Bioinformatics and dual‐luciferase reporter assays identified ZEB1 and ZEB2 as the direct targets of miR‐200 family.

CONCLUSION

Our results define a novel oncogenic regulatory mechanism of specific microRNA group in patient derived cholangiocarcinoma xenograft models and suggesting that miR‐200 cluster may be the molecular targets for eradication of human cholangiocarcinoma.

Support or Funding Information

TGEN grant from Translational Genomics Research Institute

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 Art of Border Crossings: Integrative Multidisciplinarity in the Natural Sciences and the All Atom Model of a Native HIV Capsid
Authors: Gronenborn Angela
Published: June 2021

Those of us engaged in active experimental research careers rarely have the opportunity to step back from the lab bench or the computer to reflect on how scientific practice has changed, what kinds of interactions have proved effective, and how to begin to articulate the features of successful research. In the field of Structural Biology, which is in practice deeply inter‐ and multi‐disciplinary, research at the interfaces of traditional disciplines, such as Biology, Chemistry, and Physics is essential for addressing the most important problems. If future progress in the natural sciences depends on escaping from the restraints of narrowly disciplinary language and frameworks, as I believe it is, then we will need new approaches and language to negotiate these boundaries and teach the ‘art of border crossing’ to young scientists. In my talk I will present an example of research carried out by a team of scientists that hail from different fields. Using solution NMR, MAS solid state NMR, crystallography, cryoEM and large‐scale MD simulation permitted the construction of all‐atom models for the hexamer‐of‐hexamer and pentamer‐of‐hexamer elements that make up the HIV‐1 capsid core, which, in combination with cryo‐ET, allowed us to derive a realistic all‐atom model for the entire capsid.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Personalizing Food to the Gut Microbiome
Authors: David Lawrence
Published: June 2021

Dietary polysaccharides from plants nourish human gut bacterial communities (microbiota). These carbohydrates promote the growth of resident microbes that resist pathogens, metabolize drugs, and train the immune system. Short‐chain fatty acids, which are end products of microbial polysaccharide fermentation, are also crucial metabolic precursors and energy sources for human colon cells. Yet, despite growing awareness for the importance of polysaccharides on gut bacterial growth and metabolism, optimal strategies for consuming these carbohydrates have lagged. Multiple sources of dietary carbohydrates (prebiotics) exist and each has varying effects on gut bacterial species. Moreover, gut microbial communities vary widely between people and even within individuals over time. Here, we describe our efforts to understand how prebiotic treatments should account for inter‐individual differences in gut microbiota composition. These efforts rely on experimental tools including high‐throughput bacterial culture assays and artificial gut systems. Using these systems, we find evidence for prebiotic‐ and individual‐specific components of gut microbiota responses to prebiotic therapy. We also observe changing prebiotic responses within individual gut microbiota over time. Lastly, our experiments suggest the potential for developing diagnostics to predict individual prebiotic response.

Support or Funding Information

LAD acknowledges support from the Global Probiotics Council, a Searle Scholars Award, the Hartwell Foundation, an Alfred P. Sloan Research Fellowship, the Translational Research Institute through Cooperative Agreement NNX16AO69A, the Damon Runyon Cancer Research Foundation, the Hartwell Foundation, and the NIH (1R01DK116187‐01, P30 DK034987).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cell line xenograft growth in the B6;129‐Rag2tm1FwaIL2rgtm1Rsky/DwlHsd (R2G2) mouse model
Authors: Naden Jamie
Published: June 2021

Immunodeficient mouse models are helping to advance the field of oncology. A new model on the market, the B6;129‐Rag2tm1FwaIL2rgtm1Rsky/DwlHsd (R2G2) knockout mouse, lacks responsiveness to common gamma chain cytokines, including IL‐2, IL‐4, IL‐7, IL‐9 and IL‐15. In addition, this model exhibits defects in lymphoid development and so lacks mature lymphocytes of the B, T, and natural killer (NK) cell lineages. Herein we describe growth and take rate of a few tumor cell line xenografts in the R2G2 immunodeficient mouse model. The cell line xenografts studied include multiple myeloma, pancreatic, Burkitt's lymphoma, ovarian, and breast cancer. Multiple myeloma (MM1.S cells) were inject via the tail vein at 50 × 104 cells per mouse. IVIS imaging was used to measure tumor burden of luciferase transfected cells. The pancreatic cancer (BxPC‐3 cells) was injected subcutaneously at 1 × 106 and tumor volume was measured weekly. Burkitt's lymphoma (RAJI) cells, ovarian (A2780) cancer cells, and breast (T47D, MDA‐MB‐231, and MCF‐7) cancer cells were implanted subcutaneously at 1 × 106 cells in both flanks of five mice each. Tumor volume and body weight was measured twice weekly for five weeks. These data provide evidence that the R2G2 mouse model is a valuable tool for oncology programs including cell line tumor models research, with high take rates and quick growth of allogeneic models.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Rare Case of an 83‐Year‐Old Male Transgender: An Anatomical and Genetic Study
Authors: Polcaro Lauren , Lawson Craig , Tan Yun , Tersigni‐Tarrant MariaTeresa , Frolov Andrey , Martin John
Published: June 2021

Background

Transgender is a broad term which describes an individual whose gender self‐identification is different from the assigned sex at birth. When the latter becomes overwhelming and causes a significant psychological impact, the term gender dysphoria is used thereby presenting itself as an extreme case of transgenderism. The individuals with gender dysphoria often seek a medical intervention, surgical or therapeutic, to correct the existing discordance between their gender self‐identification and physical appearance.

Results

In the current report, we describe a rare case of an 83‐year‐old male who self‐identified as a female by legally changing his first and middle names to female ones and whose death certificate states his sex as a female. Yet, an initial visual examination of the body revealed an absence of penis with a large scrotum and atrophied testes. It was an opinion of an experienced urologic surgeon that this individual most likely underwent a penectomy with perineal urethrostomy. Dissection of the pelvic cavity and perineum revealed normal male anatomy with testes, spermatic cords, ductus deferens, seminal vesicles, and prostate intact. Histological analysis of the testes and prostate revealed striking anomalies within the tissues. The seminiferous tubules had an abnormal epithelium with no germ, Sertoli, or Leydig cells present. Histological analysis of the prostate pointed to its non‐functional state whereas the gross tissue structure was more consistent with a prostate of a young adult. Yet amyloid deposits were found in the vasculature in the testes and prostate, which would support sepsis being the official cause of death. In order to gain additional insights into the current case, a genetic screen was performed by exome sequencing of DNA extracted from the embalmed cadaveric tissue using the Next Generation Sequencing (NGS) Illumina platform

Conclusion

The available personal information pertinent to the examined body identifies this individual as a transgender. The results of the genetic screen will help to define this case further by establishing a probable cause for penectomy and choosing between two most obvious alternatives; an elective surgery for the correction of gender discordance or a surgery associated with penile or urethral cancer. Yet, the genetic screen could also indicate an impaired hormonal production and/or signaling that would explain the observed testicular and prostatic atrophies. If not, the latter could be caused by a chemical castration. Overall, all those data will help to assess the severity of the current case of transgenderism and its possible progression to gender dysphoria.

Support or Funding Information

Center for Anatomical Science and Education, Department of Surgery, Saint Louis University School of Medicine

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Anti‐hypertensive Treatment Lisinopril on Anxiety‐like Behavior in Adult Hypertensive Dahl Salt‐Sensitive Rats
Authors: Fenner Rachel Claire , Grigorova Yulia , McDevitt Ross , Petrashevskaya Natalia , Wei Wen , Lakatta Edward , Fedorova Olga
Published: June 2021

Background

Independent of a high salt intake, Dahl Salt‐Sensitive (DSS) rats demonstrate an age‐associated increase in blood pressure (BP) accompanied by cognitive decline. We sought to explore the effect of anti‐hypertensive treatment, Lisinopril, which lowers tissue angiotensin II levels, on BP and behavior in adult DSS rats. We hypothesized, that Lisinopril, in addition to BP decrease, will affect the cognitive status of adult DSS.

Methods

Sixteen 6‐month‐old male DSS rats were kept on normal salt diet for the duration of the study; eight of them were continuously treated with 250 mg/L Lisinopril administered through their water for 2 months (15 mg/kg/day), and eight DSS rats remained on regular water as a control. Systolic BP (SBP) was measured in conscious male DSS rats via tail‐cuff plethysmography, and open field test (OFT) was performed to assess locomotor and anxiety‐like behavior at baseline prior to treatment and repeated 2 months after treatment began. Statistical regression analyses were performed using Prizm 7: 1‐way and 2‐way ANOVA, T‐test when applicable. The data are presented as mean ± standard deviation.

Results

Prior to treatment, all DSS rats displayed hypertension and BP elevation with age (SBP: 170 ± 11 vs. 142 ± 5 mmHg, 6‐mo vs. 3‐mo, n=16; P<0.01). After 2 months, control untreated animals showed no difference in SBP and were hypertensive (170 ± 15 vs. 168 ± 13 mmHg, 8‐mo vs. 6‐mo, n=8; n/s). Rats treated with Lisinopril for 2 months showed a dramatically decreased SBP compared to their baseline prior to treatment (119 ± 9 vs. 171 ± 11 mmHg, P<0.0001, n=8; Fig. 1), verifying the efficacy of the drug. In OFT, treatment with Lisinopril increased exploration of the center of the field compared to the control group (160 ± 45 vs. 91 ± 55 % of distance traveled in center to baseline, p=0.0166, n=8/group; Fig. 2) without affecting total distance traveled, indicating a reduction in anxiety‐like behavior. Furthermore, there was a significant negative correlation between SBP and increased activity in the center of the field (Pearson R=−0.58, R2=0.34, P=0.018; Fig. 3), suggesting anxiolytic effects of BP lowering medication.

Conclusion

Lowering of BP is associated with reduced anxiety‐like behavior in DSS, which indicate the connection of cardiovascular disorder and cognitive function in adult hypertensive DSS. Whether Lisinopril has an additional direct effect on brain functions in DSS rats will be further studied.

Support or Funding Information

Supported by NIH/NIA Intramural Research 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.
Direct Aortic Origin of the Left Vertebral Artery in Two Adult Female Cadavers
Authors: Donohue James , Memar Shayan , Tan Yun , Daly Daniel , Martin John
Published: June 2021

Background

The vertebral artery (VA) is a branch of the aortic arch which usually arises from the posterosuperior aspect of the first part of the subclavian artery and ascends medially to the transverse process of the seventh cervical vertebra and continues ascending through the transverse foramina of the upper six cervical vertebrae to enter the skull via the foramen magnum. It is an important blood supply of the spinal cord, vertebral muscles, hindbrain and contribution to the circle of Willis.

Results

During routine dissection of the superior mediastinum and neck of two adult female cadavers the left VA was observed originating directly from the aortic arch rather than the left subclavian artery. In one cadaver the left VA entered the transverse foramen of the fifth cervical vertebra (CV5), while in the other cadaver it entered the typical the sixth cervical vertebra (CV6) transverse foramen. The diameter and length of the vessel that entered CV5 maintained the same diameter of 5mm from its origin to its position on the posterior arch of the atlas and measured 95mm in length from its origin to its entrance at CV5. In the other cadaver, the diameter decreased from 4mm, at its origin, to 3mm as it entered CV6. As it ascended from its origin it took a highly coiled approach and measured 60mm in length from its origin to it entrance in CV6. Direct aortic origin of the left vertebral artery is the most frequent anatomic variant of the vertebral artery with an incidence ranging from 2.4–5.8%.

Conclusion

Knowledge of this variation is important to reduce the risk of vertebral artery injury in angiography and surgical approaches.

Support or Funding Information

Center for Anatomical Science and Education, Department of Surgery, Saint Louis University School of Medicine

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Digital tools and software in a blend with plastinated specimens in the 21st‐century Anatomy classroom
Authors: Stashenko Vetaley
Published: June 2021

During the last decade, anatomical education has undergone dramatic changes related to the form and especially, a pedagogical approach to the curricular content of the classroom presentations. Active learning is one of the valuable tools for didactic instruction and start actively utilize 3D software and plastinated specimens for the expansion of the classroom beyond college campus boundaries. 3D software and plastinated specimens have numerous advantages in comparison to the traditional classroom learning style. Data from the assessment of n=86 student volunteers are included. A cross‐over study was designed to compare the traditional classroom style and utilization of 3D software and plastinated models. Collected data shows that the new approach leads to more than double (218 % p< 0.001) of the students study time (volunteer homework), improvement of quiz scores (24.5% p< 0.009), assessment of self‐confidence before the class quiz (46.8% p<0.005), self‐assessment of improvement of terminology spelling and pronunciation (78% p<0.004) and occurrence of peer teaching was doubled (100% involvement p<0.007).

The study presents convincing evidence in favor of using 3D software and plastinated specimens for the delivery of anatomical curriculum materials in a more organized and effective way in situations of class time restrictions. Specifically, this methodology can be very effective with the increment of restrictions on dissection space regulations, shortage of dissection specimens and much stricter regulations on the disposal of biological waste materials

Support or Funding Information

Palm Beach State College, FL.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Animate! Breathing new life into an old pedagogy: Human embryology teaching reenvisioned
Authors: Oyedele Olusegun , Calibaba Brittany
Published: June 2021

The teaching of human embryology is as ancient as that of human anatomy itself. Over the centuries up to the present time, embryology pedagogy has evolved from enlisting the dissection of fetal cadavers and use of two‐dimensional (2D) sketches and diagrams, to the modern technologies of 2D and 3D animations, CT scans of embryos and fetuses with 3D rendering of the images, and the use of ultrasound imaging. These techniques seek to balance accurate portrayal of embryonic structures with the cognitive load associated with learning the concepts presented using these images, and their value as learning adjuncts for students. This paper discusses the rationale, pedagogical basis and implementation of narrated 2D animations of embryonic development, which present the early stages of human life in accessible cartoon‐like sequences. In response to the demand from curriculum leaders at the University of British Columbia (UBC) medical school for high‐quality, simplified yet accurate rendition of early embryonic development, we have developed the first of a series of 2D animations, which presents key embryonic processes during the first 12 weeks of life contemporaneously. First and second year MD students at UBC will have access to these animations as adjuncts to the embryology components of the MD Undergraduate pre‐clerkship program. The animation was developed using Adobe Animate® software during approximately 300 hours of programming. Audio design, recording and editing were completed with Adobe Audition®, After Effects® and Adobe Premiere Pro®. In evaluating the animation, students rated it highly for visual appeal and helpfulness to learn basic embryology, while providing useful comments on how it may be improved.

Support or Funding Information

Southern Medical Program, UBC Faculty of Medicine

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