bluuebunny/crossref_metadata_2025 huggingface dataset:
P272 How well are conventional DMARDs tolerated in PsA: a real-world study
Authors: Jawad Issrah , Nisar Muhammad K
Published: April 2020

Background

NICE guidelines recommend the first line use of DMARDs in psoriatic arthritis (PsA). However, studies show that many conventional treatments like methotrexate are poorly tolerated. There is hitherto no published real-world data addressing the tolerability of DMARDs in PsA. Our objective was therefore to assess the drug management in PsA with focus on tolerability and the reasons for therapy cessation.

Methods

We conducted a retrospective analysis of all PsA patients enrolled in electronic database up to April 2019 at our university teaching hospital. We had access to full patient records including details on co-morbidities, drugs and disease management.

Results

335 patients were identified with a formal diagnosis of PsA. Mean age of the cohort was 46 years (13-81) and 58% were female. 9% of the individuals had diabetes and 18% had concurrent cardiovascular disease. 1/10th reported to be current smokers and 8% had a diagnosis of depression. 48% of the group had clinically active disease. Same percentage were taking a single DMARD. 10% had trialled 3 or more drugs. 62% of patients had discontinued one or more DMARDs prior. The mean duration before discontinuing a DMARD was 9.9 months. Methotrexate was the best tolerated and on average discontinued after 13.4 months (range: 4 days to 10.9 years). Sulfasalazine and hydroxychloroquine were discontinued after an average of 8.4 (11 days to 4.27 years) and 12.5 months (1.3 months to 2.88 years) respectively. Leflunomide was the least tolerated DMARD and stopped after an average of 5.5 months (7 days to 2.53 years). The main reason for stopping a medication was gastro-intestinal symptoms which accounted for 42% of all the reported side effects. This applied to both methotrexate (43%) and sulfasalazine (46%) discontinuation. The leading reasons for discontinuing hydroxychloroquine were jointly GI symptoms and other side effects at 43% each. Leflunomide was stopped in 50% of cases due to neurological symptoms.

Conclusion

To our knowledge, this is the first report confirming poor retention rate of oral DMARDs in a real world PsA cohort managed over 20 years. In the context of chronic disease, the median duration of treatment is short. Our analysis did not include patients who suffer from side effects but continue therapy thereby impacting treatment adherence and hence the true scale of the issue is likely higher. Though NICE guidelines stipulate the need of an adequate trial of minimum two DMARDs prior to therapy escalation, these drugs are not well tolerated and thus pose a challenge to clinicians. One potential solution is earlier adoption of biological therapies, which are increasingly cost effective and have been shown to be better tolerated.

Disclosures

I. Jawad None. M.K. Nisar None.
P280 Clinical burden of concomitant joint disease in psoriasis: a US-linked claims and electronic medical records database analysis
Authors: Tillett William , Skornicki Michelle , Prince Patricia , Suruki Robert , Lee Edward , Louder Anthony
Published: April 2020

Background

Psoriasis (PSO) is an inflammatory skin condition, impacting physical and psychological well-being. PSO patients may develop psoriatic arthritis (PsA), a painful joint disease that further impacts quality of life. The objective was to examine rates of metabolic disease and depression in patients with PSO and PsA.

Methods

Claims data linked to electronic medical records were examined (Jan-2007-Mar-2018). Prevalent PSO patients were followed until end of data collection and categorized based on development of PsA (PSO+PsA cohort; defined by ICD9/10 codes in patients without PsA at baseline). Follow-up rates of comorbidities, including metabolic disease and depression, were compared for those with/without PsA, presented as cases per 1,000 patient-years (1000PY).

Results

Progression to PsA occurred in 2,311 (12%) patients. Follow-up rates of hyperlipidemia, hypertension, diabetes, and cardiovascular disease in the PSO+PsA cohort were 1.4-times (326 vs 234/1000PY), 1.6-times (357 vs 228/1000PY), 1.7-times (119 vs 72/1000PY), and 1.4-times (71 vs 50/1000PY) that observed in the PSO-only cohort, respectively. In patients without baseline metabolic disease, higher rates in PSO+PsA vs PSO-only remained (approximately 1.5-times and 1.3-times higher rates of hypertension and hyperlipidemia, respectively). The rate of depression was 140 vs 92/1000PY in the PSO+PsA vs PSO-only cohort; when examined in patients without baseline depression, rates were about half that observed in the overall population (72 vs 49/1000PY).

Conclusion

PSO+PsA patients had a substantial clinical burden, characterized by 40-70% higher rates of metabolic disease and depression than for patients who did not progress to PsA.

Disclosures

W. Tillett: Consultancies; AbbVie, Celgene, Janssen, Lilly, Novartis, Pfizer and UCB Pharma. Grants/research support; AbbVie, Celgene and Lilly. M. Skornicki: Other; Employee of Aetion. P. Prince: Other; Employee of Aetion. R. Suruki: Other; Employee of UCB Pharma. E. Lee: Other; Employee of UCB Pharma. A. Louder: Other; Employee of Aetion.
P282 Outcome for PsA biologic switchers following primary inefficacy of TNFi therapy in a secondary care cohort: sequential TNFi versus switching mode of action
Authors: Naja Meena , Santos Liliana R , Shipa Muhammad , Mandy Greenwood , Castelino Madhura
Published: April 2020

Background

TNFi are the most used first line of biologic disease modifying anti-rheumatic drugs (bDMARDs) in PsA. In primary loss of response (PLR) to TNFi, there is no current evidence that directs the choice of second line bDMARDs. Our aim in this work was to compare drug survival for those who switched to a second TNFi versus alternative agents in real life.

Methods

Data was analysed retrospectively from a cohort of 400 PsA patients followed through from 2002-2019. Statistical analysis was carried out with descriptive statistics and t-test analysis using SPSS version 22.

Results

Out of 400 patients, 220 (55.0%) were started on bDMARD treatment. Of these 220, 212 (96.5%) were started on TNFi as initial therapy. PLR was seen in 42 of these patients (19.8%). The median drug survival of initial TNFi therapy was 7.1 months (interquartile range [IQR] 3.6 - 53.4 months). Of the 42 patients with PLR: 32 (76.2%) were switched to a second TNFi; 6 (14.3%) were switched to ustekinumab and 4 (9.5%) were switched to secukinumab. 21 of the 32 patients switched to a second TNFi were subsequently switched onto a third biologic due to treatment failure (65.6%). The median drug survival of the second TNFi in this group was 7.7 months (IQR 3.0 - 26.3 months). 3 out of the 6 (50%) patients who were switched to ustekinumab were then subsequently switched, this was due to primary inefficacy in 66.7% and adverse events in 33.3%. The median drug survival of ustekinumab in this group was 10.0 months (IQR 0.0 - 16.0 months). All 4 patients switched to secukinumab continue on this treatment with no drop outs, giving a median drug survival 12.3 months (IQR 6.5 - 19.5 months).

Conclusion

Our data suggests that patients with PLR to TNFi in PsA who switched mode of action to IL-17 inhibitor appeared to have better drug survival than subsequent TNFi or IL-12/23 inhibitors. Exploring the clinical biomarkers for those with successful switch to non-TNFi bDMARD in a larger cohort would help with targeting the most appropriate individuals and early disease control.

Disclosures

M. Naja None. L. R. Santos None. M. Shipa None. G. Mandy None. M. Castelino None.
Fraud exception to privilege: Addlesee & Ors v Dentons Europe LLP [2020] EWHC 238 (Ch)
Authors: Munby Thomas
Published: April 2020

Some months after Addlesee & Ors v Dentons Europe LLP produced an important judgment of the Court of Appeal in relation to the statue of legal professional privilege following the dissolution of a corporate client,1 the same litigation has given rise to a judgment addressing another area of the law of privilege.
Determination of plasma-chlortetracycline (CTC) concentrations in grazing beef cattle fed one of four FDA approved free-choice CTC-medicated minerals
Authors: Reppert Emily J , Reif Kathryn E , Montgomery Shawnee R , Magnin Geraldine , Zhang Yuntao , Martin-Jimenez Tomas , Olson K C , Coetzee Johann F
Published: April 2020

Control of active bovine anaplasmosis in the United States is predicated on the use of chlortetracycline (CTC)-medicated feed throughout the vector season. However, data describing population pharmacokinetics of chlortetracycline in cows, on pasture, having free-choice access to CTC-medicated mineral for consecutive months is lacking. This study documented plasma-CTC concentrations in grazing cows during peak vector season in an anaplasmosis endemic herd. Each pasture was administered one of the four Food and Drug Administration approved CTC-medicated mineral formulations and were assigned as follows: 0.77 g/kg, Aureo Anaplaz C700 Pressed (Sweetlix Livestock Supplements, Mankato, MN); 5.5 g/kg, Purina Anaplasmosis Block (Purina Animal Nutrition, Gray Summit, MO); 6.6 g/kg, Stockmaster Aureo FC C6000 Mineral (Hubbard Feeds, Mankato, MN); 8.8 g/kg, MoorMan’s Special Range Minerals AU 168XFE (ADM Animal Nutrition, Quincy, IL). Blood samples were collected monthly for determining plasma drug concentration by Ultra performance liquid chromatography (UPLC) and mass spectrometry. Continued plasma-CTC monitoring allowed for characterization of trends between treatment groups (pastures), age groups (<3 yr or >4 yr), and sampling times (June to October). Results indicate formulation (pasture) and time were significant factors affecting concentrations of CTC in plasma. Cows exposed to 5.5 g/kg block formulation recorded higher CTC plasma concentrations compared with other pasture groups (P = 0.037). Plasma-CTC concentrations increased over time (month of measurement; P = 0.0005). Specifically, concentrations measured after 5 months of continuous CTC treatment were higher than those measured in earlier months.
Collagen VI (CVI) antibodies regulate macrophages differentiation through FcgammaRIIb signal transduction pathway
Authors: Li Zhonghao , Zhao Ming
Published: April 2020

We previously reported that monoclonal antibody against collagen VI (CVI mAb) regressing atherosclerotic plaque through inducing the shift of macrophage differentiation, namely from M1 to M2, and ensuing the macrophages engulf the plaque lipids and transfer the lipids to HDL through restoring lipid transfer protein ABCA1 expression. Here we report the cellular signal transduction on the CVI mAb inhibition of oxLDL‐induced M1 differentiation. Using purified human CD14+ cells, we find that oxLDL induces M1 differentiation is JNK and ERK MAPKs dependent. And CVI mAb inhibits M1, while activates M2 differentiation, though oxLDL also activates M2 differentiation in some way, but the M1/M2 ratio are significantly reversed by CVI mAb. The molecular mechanism behind these is that CVI mAb activates macrophage FcgammaRIIb and its downstream SHIP and Dok phosphatases, and inhibits Syk and Ras signal transduction pathway, which are MAPKs upstream kinases, so that CVI mAb inhibits ERK, JNK, and p38 MAPKs activation, and ensuing M1 differentiation. Thus, we found the signal transduction pathway of CVI mAb inhibition of oxLDL‐induced M1 differentiation: CVI mAb→FcgammaRIIb→SHIP→Dok→Syk and Ras→ERK and JNK MAPKs→M1. These results might give a new clue on control the lipids‐induced inflammatory reaction on the large blood vessel and therapeutic method to prevent atherosclerosis.

Support or Funding Information

This work was supported by the National Natural Science Foundation of China (grants 81772074, 81974045), the Natural Science Foundation of Guangdong, China (grant 2012B050600002), the Guangdong Province Talent Recruitment Foundation and the Guangdong Innovative Research Team Program (No. 201001Y0104675344), Project of Guangzhou city scientific plan (201804020002).
Anti‐tumor activity of the ethanolic extract of Micromeria fruticosa on human breast and colon cancer cells
Authors: Abu-Gharbieh Eman , El-Huneidi Waseem , Shehab Naglaa G. , Bajbouj Khuloud , Vinod Arya , El-Serafi Ahmed , Malhab Lara Bou , Abdel-Rahman Wael M.
Published: April 2020

Micromeria fruticosa (L.) Druce subsp. Serpyllifolia (Lamiaceae) is a perennial, aromatic shrub that usually distributed in the Mediterranean regions. The aerial parts of the plant are widely used to treat many diseases, including abdominal pains, diarrhea, colds, eye infections, heart disorders and wounds. A few reports have confirmed various therapeutic potentialities of its extract, such as anti‐inflammatory, gastroprotective, analgesic activities, antiobesity, and antidiabetic activities.

Aim

To investigate the mechanistic pathway for the antitumor activity of the ethanolic extract of M. fruticosa on two different cancer cell lines, namely human breast adenocarcinoma (MCF‐7), human colorectal cancer (HCT‐116) cell lines.

Methods

Cell viability assay, DNA profile with PI staining, Annexin‐V/PI apoptosis assay, microarray analysis, qRT‐PCR were used to assess the effect of M. fruticosa on cytotoxicity, cell cycle proliferation, apoptosis, and genes expression profile in treated cell lines.

Results

M. fruticosa is found to exhibit potential and effective cytotoxic activity against MCF‐7 and HCT‐116 cancer cell lines by attenuating cell proliferation and inducing cell cycle arrest. Microarray analysis revealed a significant reduction in cdk1, cyclin B1 and survivin levels upon M. fruticosa treatment. The qRT‐PCR confirmed the downregulation of cdk1, cyclin B1 that correlates with the observed G2 arrest in both cell lines. Moreover, results showed that the decreased levels of the cell death inhibitor, survivin is correlated to the promoted cell death in the treated cell lines.

Conclusion

The findings suggest promising antiproliferative potential Micromeria fruticosa in breast and colon cancers. Further studies are needed to characterize the potential active constituent.

Support or Funding Information

This research was funded by University of Sharjah Seed Grant no.1701090113‐P and UOS‐Boehringer Ingelheim award.
Nicotine exposure during pregnancy alters the maternal gut microbiome and both cecal and plasma short chain fatty acids in Sprague Dawley rats.
Authors: Hayward Linda , Watkins Jacqueline , Bautista Byrell , Lin Cindy , Malphurs Wendi , Zubcevic Jasenka
Published: April 2020

Children of mothers that smoke during pregnancy are more likely to be obese and develop high blood pressure, suggesting a developmental origin of these diseases. Evidence now suggests that an imbalance of the bacterial microbes in the gastrointestinal tract may also play an important role these diseases. Thus, the present study was undertaken to test the hypothesis that nicotine‐exposure during pregnancy alters the maternal gut‐microbiome, inducing gut dysbiosis that may be passed on to the child during birth and/or shifts in bacterial metabolites, such as short chain fatty acids (SCFAs), that the fetus is exposed to in utero. To test our hypothesis, age‐matched virgin and pregnant female rats were exposed to nicotine (6 mg/kg/day) or vehicle (saline) via osmotic mini pump, including days 6–18 of pregnancy. Cecal contents, tissue from the proximal colon, and plasma were collected on day 14 of treatment. A two‐way ANOVA (pregnant state x treatment) identified that pregnancy decreased the abundance of bacteria in the phyla Firmicutes, while increasing the abundance of Actinobacteria and Verrucomicrobia compared to virgin rats (P<0.02). Nicotine exposure, independent of pregnancy, further increased the abundance of Actinobacteria (P<0.03) and tended to increase the proportion of Firmicutes and decrease the abundance of Verrucomicrobia. Evaluation of cecal SCFAs identified that pregnancy upregulated the abundance of acetate and propionic acid (P<0.02) and tended to upregulate butyric and hexanoic acid. Conversely, nicotine exposure, independent of pregnancy, decreased cecal acetate (P<0.02), propionic acid (P<0.007), and hexanoic acid (P<0.009) concentrations. Similarly, plasma levels of acetate, propionic, butyric, and valeric acid were upregulated by pregnancy, and nicotine exposure, independent of pregnant state, reversed this trend (P<0.05). Finally, gene expression in the proximal colon identified that pregnancy triggered an upregulation of multiple genes, including the protective enzyme 11‐beta HSD2, the sodium channel ENAC, Ffar2, the SCFA acetate and propionate transporter, the solute carrier SMCT1, and Tph2, the rate limiting enzyme for serotonin production (P<0.05). Independent of pregnancy status, nicotine downregulated the expression of all five genes. These results confirm our hypothesis that nicotine exposure during pregnancy can significantly alter the maternal gut microbiome and affect fetal exposure to specific circulating factors linked to gut microbial metabolism. These findings provide a new link between changes in the maternal gut microbiome and the fetal origins of adult diseases.

Support or Funding Information

FL DOH James and Esther King Biomedical Research Program 8JK06
Neuroinflammation and Microglial Activation in The Central Amygdala in Rats with Heart Failure
Authors: Althammer Ferdinand , Neto Hildebrando Candido Ferreira , Stern Javier Eduardo
Published: April 2020

Cardiovascular diseases (CVDs), such as heart failure (HF) are the most common cause of death globally. Moreover, clinical studies support a high degree of comorbidity between CVDs cognitive impairment, and emotional distress, with 20–40% of all HF patients displaying depression and anxiety levels significantly higher than the healthy population. Still, the precise underlying mechanisms causing those cognitive and emotional impairments remain elusive. The paraventricular nucleus of the hypothalamus (PVN) is a key brain structure involved in emotional control and directly communicates with the central lateral part of the amygdala (CeA), which actively participates in the regulation of emotion and fear memories. While markers of neuroinflammation (NI) have been previously described in the PVN of HF rats, it is at present unknown whether the CeA is also affected. To address this question, we used an ischemic HF rat model (ejection fraction <35%). We developed a comprehensive quantitative microglial profiler using the IMARIS software along with immunohistochemical staining with the microglial marker IBA‐1 to assess changes in various microglial parameters indicative of different microglia activity stages. At early stages of the established HF (8wks post‐surgery), we found elevated indexes of microglial activation in the PVN compared to sham controls (15.4% decrease in surface area, 29.9% decrease in cell volume, 15.5% decrease in filament length and 35.1% increase in IBA‐1 immunoreactivity intensity, p<0.001 in all cases). These changes became more pronounced at later (14wks) stages of the disease. While no changes were observed in the CeA at the early stage, significant changes were observed at 14wks after HF compared to sham controls (21.2% increase in cell volume, 9.7% decrease in filament length and 12.3% increase in IBA‐1 immunoreactivity intensity, p<0.05 in all cases). These morphometric changes are indicative of de‐ramification and somatic swelling of microglial cells. These findings were corroborated by elevated mRNA levels of IBA‐1, TNF‐a, IL‐1β and IL‐6 measured via qPCR in PVN and CeA punches from HF rats. Importantly, no microglial changes were observed in cortical areas suggesting that HF‐induced NI was not a diffuse phenomenon affecting the entire brain. To the best of our knowledge, our results highlight for the first time that in addition to the PVN, HF also results in a robust microglial cell activation in the CeA, a critical forebrain region involved in emotion and cognition. NI‐related changes in the CeA occurred at a later stage of the disease, and could potentially contribute to cognitive impairment and mood disorders in HF patients.

Support or Funding Information

National Institutes of Health grants R01NS094640 and R01HL090948 to JES
Quantifying Cardiac Injury with Donation after Circulatory Death Process: Impact of Donor Age and Obesity
Authors: Akande Oluwatoyin Elizabeth , Chen Qun , Toldo Stefano , Lesnefsky Edward , Quader Mohammed
Published: April 2020

Introduction

Heart transplantation, the gold standard treatment for patients with advanced heart failure, is limited due to shortage of suitable donor hearts, which at the present are procured from Brain Death Donors (DBD) only. Recent advancement in organ procurement and retrieval techniques have shown that Donation after Circulatory Death (DCD) promises to expand the heart donor pool. While ischemic insult is inherent in the DCD process, subsequent reperfusion injury is inevitable in these hearts. Donor age and obesity may adversely affect the function of a DCD heart. We herein hypothesize that age and obesity may influence the outcome of cardiac injury in DCD hearts.

Methods

SD rats ages between 13 and 17 weeks, weighing between 340 and 430 grams were subjected to 25min of warm ischemia following the DCD protocol. Hearts from these DCD rat models were reanimated on Langendorff system primed with KH buffer at 37°C, and perfused at steady 73mmHg for 60 minutes, to obtain cardiac function (left ventricle developed pressure‐LVDP, positive and negative dP/dt) data.

Results

Following 25 min of warm ischemia in vivo, we observed lower LVDP (43±10 mmHg) in DCD hearts from rats older than16 weeks or weighing more than 370 grams when compared with LVDP ( 64±12 mmHg ) in hearts from rats younger than 16 weeks of age or weighing less than 370 grams. Similar trend was noted in positive and negative dP/dt values. The decrease in DCD heart function was near linear with both increasing age and weight (Figure 1).

Conclusions

Based on our findings, age and obesity of DCD donor influences susceptibility to warm ischemia. These donor factors need to be taken into consideration when designing studies involving DCD rat hearts.
Spatio‐Nutritional Regulation of Intestinal Desaturases
Authors: Burchat Natalie , Akal Tasleenpal , Zalma Brian , Ye Hong , Kumaraswamy Deeptha , Sampath Harini
Published: April 2020

Stearoyl‐CoA desaturases are ER‐resident enzymes that convert saturated fatty acids into monounsaturated fatty acids and have been studied using whole body, liver, skin, and adipose knockout models, but their specific role in the intestine is not well understood. Stearoyl‐CoA desaturase 1 (SCD1) gene and protein expression increases along the length of the intestine and was found to be induced by fasting followed by refeeding a high sucrose very low‐fat diet, similar to what has been established in liver. In addition to SCD1, stearoyl‐CoA desaturase 2 (SCD2) was also found to be expressed and regulated in a similar manner to SCD1 in the intestine. Intestine specific SCD1 knockout (iKO) mice were generated by crossing SCD1 floxed mice with mice expressing the Cre‐recombinase gene under the villin promoter. These mice have longer intestines and increased lipid absorption compared to floxed counterparts. In addition, iKO mice have increased bile acids, a more hydrophilic bile acid pool, and have increased plasma GLP‐1 and brown adipose tissue (BAT) Iodothyronine Deiodinase 2 (Dio2) gene expression. When placed on a low‐fat diet (10% fat) or a high‐fat diet (45% fat), the iKO mice had increased food intake and energy expenditure while maintaining similar or leaner body weights as their floxed counterparts. The iKO mice showed a trend of improved glucose tolerance. Overall, these results show that stearoyl‐CoA desaturases are expressed and regulated in the intestine. Knocking out SCD1 in an intestine‐specific manner leads to increased bile acids and these increased bile acids may contribute to the increased GLP‐1 and bile acid‐induced thermogenesis in BAT. When fed a high fat diet, iKO mice demonstrated an increased basal metabolic rate, increased food intake, and improved glucose tolerance.
Lack of Uncoupling Protein 3 Protects from High‐Fat Diet‐Induced Obesity, Systemic Inflammation and Insulin Resistance in Rats
Authors: Lomax Tyler , Ashraf Sadia , Yilmaz Gizem , Harmancey Romain
Published: April 2020

Introduction. Uncoupling protein 3 (UCP3), a mitochondrial anion carrier with antioxidant properties, is downregulated in animal models with obesity, insulin resistance and type 2 diabetes. Although mutations in UCP3 have been linked to obesity in humans, their contribution to increased adiposity and insulin resistance is still unclear. The goal of this study was to investigate the impact of UCP3 loss on the development of high‐fat diet‐induced obesity in rats.

Methods. Male UCP3 knockout (KO) rats and their wild type (WT) littermates were maintained on a high‐fat diet (45% kcal from fat) for up to 21 weeks starting at 8 weeks of age. Body composition was analyzed by EchoMRI. Whole‐body insulin sensitivity and tissue‐specific glucose uptake were determined during hyperinsulinemic‐euglycemic clamps studies. White and brown adipose tissue morphologies were assessed by histology and electron microscopy. Potential molecular alterations in brown adipose tissue (BAT) physiology were further investigated by RNA sequencing, quantification of total protein S‐sulfenylation, and determination of tissue glutathione levels.

Results. Loss of UCP3 unexpectedly decreased body weight gain on a high‐fat diet in spite of increased food intake. The decrease in body weight gain was driven by a decrease in body fat mass leading to smaller white adipocytes and a decrease in circulating tumor necrosis factor alpha levels. Consequently, the KO rats exhibited preserved insulin sensitivity and glucose tolerance which correlated with increased glucose uptake in BAT. While high‐fat feeding induced lipid accumulation and whitening of BAT in WT rats, this phenomenon was markedly reduced in KO rats. Surprisingly, protein S‐sulfenylation was decreased in BAT from KO rats, suggesting decreased mitochondrial reactive oxygen species generation in spite of UCP3 loss. The RNA sequencing analysis of BAT transcriptome revealed increased expression of multiple enzymes linked to glutathione synthesis. Direct quantification of glutathione confirmed a 44% increase of the antioxidant molecule levels in BAT of KO rats when compared to high‐fat fed WT rats.

Conclusions. Lack of UCP3 protects from high‐fat diet‐induced obesity and insulin resistance in rats. We propose that this protective effect is driven by a compensatory increase in synthesis of the antioxidant glutathione in BAT, thereby protecting mitochondrial function and substrate utilization in this tissue. The findings suggest that loss of UCP3 function in humans may unexpectedly decrease whole‐body fat accumulation, inflammation and the development of insulin resistance.

Support or Funding Information

This work was supported by grants R01HL136438, P20GM104357 and P01HL051971 from the National Institutes of Health.
Effective Reinnervation of Skeletal Muscle is Impaired by Disrupting Microvascular Regeneration Following Acute Injury
Authors: Morton Aaron B , Cornelison DDW , Segal Steven S
Published: April 2020

Acute injury of skeletal muscle damages myofibers, microvessels and neuromuscular junctions (NMJs). Although myofiber regeneration is well‐characterized, little is known of concurrent events in microvessels and motor nerves during skeletal muscle regeneration. Because growth and patterning are coordinated between nerves and microvessels during development, we tested the hypothesis that effective microvascular regeneration is required for restoring neuromuscular structure and function post injury in the adult. To impair angiogenesis, mice were bred for endothelial cell (EC)‐specific deletion of ephrin‐B2. Male mice (3–6 months old; n=2–3/group) were injected with tamoxifen to delete ephrin‐B2 in ECs (EfnB2CKO; confirmed with PCR); uninjected mice of the same genotype were used as controls. The gluteus maximus (GM) or tibialis anterior (TA) muscle was injured by local injection of 1.2% BaCl2 and muscles were evaluated at 10 days post injury, by which time morphological and functional myofiber regeneration will have occurred. Intravascular injection of fluorescent wheat germ agglutinin (WGA; stains only perfused vessels) or immunolabeling for CD31 (stains all ECs) revealed microvessels of the GM in EfnB2CKO to be vacuolated with disorganized ECs vs. intact microvascular networks with aligned ECs in control mice. Using indirect stimulation via the sciatic nerve, isometric force production in the TA was 19±6 N/cm2 in EfnB2CKO vs. 33±3 N/cm2 in control mice; force produced with direct electrical field stimulation was similar between groups. Immunostaining for neurofilament‐heavy revealed reduced presynaptic coverage of NMJs in EfnB2CKO vs. control [60±9% vs. 74±10% of acetylcholine receptors (AChRs) per 200 μm2 NMJ area]; postsynaptic staining with α‐bungarotoxin revealed dispersed AChRs [200±118 vs. 92±43 fragments/200 μm2, respectively]. These preliminary findings indicate that the loss of ephrin‐B2 in ECs results in functional deficits for neuromuscular transmission and NMJ morphology in association with impaired microvascular regeneration following acute injury. We conclude that regeneration of intact NMJs with functional reinnervation of myofibers requires effective revascularization and recovery of the microcirculation following skeletal muscle injury.

Support or Funding Information

Support: APS Postdoctoral Fellowship (ABM), R01AR067450 (DDWC), R37HL041026 (SSS)
Metabolic Effects of Whey Protein Isolate Supplementation on Cholesterol and Triglycerides in Women with Polycystic Ovary Syndrome
Authors: Balcom-Luker Shenavia , Kaiser Kayleigh , Zumbro Emily L. , Rao Manisha , Low Irene , Broughton K. Shane , LeMieux Monique J.
Published: April 2020

The current study examined the acute‐ and chronic effects of whey protein isolate ingestion on plasma cholesterol and triglyceride concentrations in women with polycystic ovary syndrome (PCOS). PCOS, an endocrine abnormality in women of reproductive age, is characterized by menstrual cycle disturbance, hyperandrogenism and obesity. Women with PCOS have altered metabolic profiles and are more likely to develop type 2 diabetes and/or liver disorders such as non‐alcoholic fatty liver disease (NAFLD). Nutritional management options are severely limited. Whey protein isolate (WPI) improves metabolic parameters in patients with and without type 2 diabetes, but its therapeutic potential in polycystic women has yet to be realized. Given the association between PCOS and NAFLD, we hypothesized that supplementation with WPI may attenuate circulating cholesterol and triglycerides in a polycystic population. Fifteen premenopausal women with PCOS and eleven healthy, regularly menstruating women underwent three 150‐min oral glucose tolerance tests (OGTT): (i) OGTT without protein preload, (ii) OGTT on 1st, and (iii) 40th day of protein preload. Participants consumed 35g protein daily with protein intake 30 min before carbohydrate load on test days. Plasma levels of cholesterol and triglycerides were assessed at baseline, day 1 and day 40 to determine acute and chronic effects on cholesterol and triglycerides using a Biolis 24i chemistry analyzer. (Participant recruitment, data collection and analyses in progress.) Protocols used in this study were approved by the TWU Institutional Review Board. Mixed Model Repeated Measure ANOVA indicates that time and condition (PCOS vs control) did not have a significant effect on cholesterol or triglyceride levels within and between groups (p > .05). However, preliminary data indicate that disease condition had a significant impact on cholesterol (p< .001) and triglyceride (p = .001) concentrations at baseline. Changes in cholesterol and triglyceride levels after 40 days of protein preload are currently being analyzed. Women with PCOS have higher levels of cholesterol and triglycreides than those without this condition. While acute effects of 35g WPI ingestion do not seem to alter these levels, a longer duration of supplementation may induce more beneficial effects (currently being conducted). Findings from this study will provide foundational support for WPI as a nutritional treatment for PCOS and reveal insights into cholesterol metabolism mechanisms that are affected by PCOS and NAFLD.

Support or Funding Information

Glanbia Nutritionals, INC; Human Nutrition Research Funds, Texas Woman’s University; Research Enhancement Program, Texas Woman’s University
Skeletal Muscle Reflex‐Induced Dysregulation of Sympathetic Nerve Activity in Type 1 Diabetic Rats
Authors: Kim Han-Kyul , Ishizawa Rie , Hotta Norio , Iwamoto Gary , Vongpatanasin Wanpen , Mitchell Jere , Smith Scott , Mizuno Masaki
Published: April 2020

The pressor response to physical exercise is exaggerated in the early stages of type 1 diabetes mellitus (T1DM). This augmented blood pressure (BP) response has been further shown to be mediated by an overactive skeletal muscle exercise pressor reflex (EPR). However, whether alterations in sympathetic responsiveness contribute to this EPR overactivity is unknown. The purpose of this study was to test the hypothesis that the potentiated BP response to exercise in T1DM is mediated by EPR‐induced sympathetic overactivity. Sprague‐Dawley rats were intraperitoneally given either 50 mg/kg streptozotocin (T1DM) or saline (control). We measured renal sympathetic nerve activity (RSNA) and mean arterial pressure (MAP) responses to activation of the EPR, as well as its functional components, the mechanoreflex and metaboreflex, in decerebrate animals at 1–3 weeks after administration. Compared to control rats, selective activation of the EPR evoked significantly greater increases in RSNA (Δ = 58±7 vs. 96±15 %, P<0.05) and MAP (Δ = 11±2 vs. 33±5 mmHg, P<0.05) in T1DM rats. The sympathetic and pressor responses to the individual stimulation of the mechanoreflex and metaboreflex were also significantly larger in T1DM animals than in control animals (P<0.05). These findings suggest that in untreated T1DM, abnormally large muscle reflex‐evoked sympathetic overactivity contributes significantly to exaggerations in BP during exercise. Importantly, the results implicate sympathetic overactivity as a potential therapeutic target for improving/preventing the abnormally high BP response to exercise in T1DM.

Support or Funding Information

The Southwestern School of Health Professions Interdisciplinary Research Grant Program, the Lawson & Rogers Lacy Research Fund in Cardiovascular Disease and the JSPS KAKENHI JP17K01769
Effect of Dietary Nicotinamide Mononucleotide (NMN) on Function and Mechanics of Skeletal Muscle Arteries from Aged Mice
Authors: Murphy Timothy V. , Wu Lindsay , Sandow Shaun L. , Das Abhirup , Naim Hussein
Published: April 2020

Ageing is associated with a decline in vascular function and increased risk of cardiovascular disease. Dietary NMN, an NAD+ precursor, increases muscle vascularity, blood flow and exercise performance in aged mice
. This study determined the effects of dietary NMN on endothelium‐dependent dilation (EDD) and vascular stiffness in skeletal muscle arteries isolated from aged mice. Male and female C57BL/6J mice received NMN (400 mg/kg) in drinking water for 8–10 weeks; mice were ~97 weeks old at the end of the treatment period. Segments of the saphenous and gracilis muscle arteries (SA and GMA respectively) were removed. Functional responses were examined using pressure myography. SA and some GMA were pre‐constricted with phenylephrine. Pressure‐induced myogenic tone was enhanced in GMA from NMN‐treated male mice. NMN treatment enhanced EDD in the GMA of both male and female mice as measured by a leftward shift in the pEC50 of acetylcholine concentration‐response curves. EDD in the SA from either sex was not altered by NMN treatment. NMN treatment also did not affect responses to the endothelium‐independent vasodilator sodium nitroprusside in either artery, from either sex. The nitric oxide synthase (NOS) inhibitor L‐NAME (100 μM) strongly inhibited EDD in the SA but caused only minor inhibition of EDD in the GMA, from both sexes. In the GMA from both sexes, EDD was significantly inhibited by blockers of Ca2+‐sensitive K+ channels (KCa). Analysis of relative EC50‐shift suggested intermediate‐conductance‐KCa (IKCa)‐mediated vasodilation was enhanced in the GMA from NMN‐treated mice. Stress‐strain relationships were obtained from arteries under passive conditions, over the pressure‐range 5–120 mmHg. Analysis of these relationships showed NMN treatment increased compliance of GMA, but not SA, from both sexes. SA from female mice were less compliant than SA from male mice, regardless of NMN treatment. These studies suggest NMN treatment can enhance EDD, decrease arterial stiffness and improve blood flow autoregulation in small resistance (GMA) arteries, but not larger conduit (SA) arteries in skeletal muscle from aged mice. The increased EDD may be due to increased IKCa‐mediated vasodilation, but not increased nitric oxide‐mediated effects. These changes may contribute to the enhanced muscle blood flow and exercise performance observed in aged NMN‐treated mice.
Calorie Restriction Delays the Aging of Saccharomyces cerevisiae by Improving Complex III Activity via Glutathione Peroxidase 2 (Gpx2)
Authors: Hurtado-Villagómez Ana Victoria , Olmos-Orizaba Eridani , Saavedra - Molina Alfredo , Rodriguez-Orozco Alain Raimundo , Calderon-Cortes Elizabeth , Cortes-Rojo Christian
Published: April 2020

Comparative studies among organisms of different species of the same genus have found that long ‐ lived species contains polymorphisms in the sequence of the cytochrome b (Cyt b) subunit of the complex III leading to lower ROS production, which has been suggested to contribute for longer lifespans. Cyt b is a highly hydrophobic protein that depends on cardiolipin for its function. Cardiolipin peroxidation impairs electron transfer at the cytochrome b and enhances ROS by inducing electron leak. Cardiolipin peroxidation is repaired in mitochondria by Gpx2. Calorie restriction, the only non‐genetic manipulation increasing the lifespan, induces a hormetic response enhancing the activity of antioxidant systems like Gpx. Thus, it is feasible to hypothesize that calorie restriction increases longevity in yeast by preserving complex III activity and electron transfer in the Cyt b in a Gpx2 – dependent manner. Calorie restriction was induced in yeast with 0.05% glucose in YPD medium. YPD 0.05% extended the chronological lifespan of yeast in comparison with YPD medium with standard 2% glucose. Calorie restriction also improved complex III activity in aged yeasts and enhanced the reduction of Cyt b. The beneficial effects of calorie restriction on chronological lifespan, complex III activity and Cyt b reduction were lost in a GPX2Δ yeast mutant. These results suggest that preservation of complex III activity by enhanced electron transfer at Cyt b might be another mechanism by which calorie restriction extends longevity, which might be related with lower ROS generation and protection of cardiolipin against oxidative damage induced by aging.

Support or Funding Information

This work was supported by a grant from Programa de Investigación 2019, Coordinación de la Investigación Científica ‐ UMSNH (CCR).
Effects of the Bioflavonoid Quercetin on TLR4 Expression and NFKb activation in A375 Melanoma Cells
Authors: Hawley Cassidy , Mankins Claire , Byrd Shere K.
Published: April 2020

Early research indicates that quercetin, a bioflavonoid, seems to limit cancer progression through the down regulation of toll‐like receptor 4 (TLR4). Many cancers, including melanoma, upregulate expression of TLR4 which appears to be associated with cell survival and proliferation. The purpose of this study is to examine TLR4 expression, downstream signaling and changes in target gene expression that may contribute to quercetin’s anti‐proliferative properties. Cells were treated with 100 uM quercetin, vehicle control (DMSO), or no treatment for 24 hours. NFKb activation was quantified by differential centrifugation of cytosolic and nuclear fractions of treated cells by immunoprecipitation and Western blotting. TLR4 was evaluated with immunoblotting. We found that quercetin treated cells had 15% higher NFKb nuclear fraction than the cells with no or vehicle only treatment. TLR4 was, however, effectively downregulated by quercetin treatment. These findings are similar to those seen pancreatic cancer and some other cell models. It appears that this bioflavonoid may have different targets and effects in different types of cancer cells, making it necessary to understand its differential mechanisms of action.

Support or Funding Information

Supported by the NIH Common Fund, through the Office of Strategic Coordination, Office of the NIH Director with the linked awards: TL4GM118992, RL5GM118990, UL1GM118991 and by Fort Lewis College MARC‐USTAR.
Bile Acid Membrane Receptor TGR5 Inhibits Alcohol‐Induced Liver Steatosis
Authors: Fan Mingjie , Liu Yanjun , Tu Jui , Wang Yangmeng , Jin Lihua , Huang Wendong
Published: April 2020

Alcoholic liver disease (ALD) is one of the leading causes of liver failure in the world. It progresses from alcoholic fatty liver (AFL) to alcoholic steatohepatitis (ASH), which is characterized with hepatic inflammation. Chronic ASH will result in fibrosis, cirrhosis and hepatocellular cancer (HCC) eventually. However, very few treatment options for ALD are available currently. Therefore, there is an urgent unmet medical need to better understand the molecular mechanisms underneath the development of ALD and develop effective treatment approaches. Bile acid signaling in the liver‐gastrointestinal tract axis is important for the integrated regulation of lipid, glucose, and energy metabolism. Dysregulated bile acid signaling is associated with ALD due to the fact that alcohol significantly alters bile acids pool and composition in humans and rodents. G‐protein‐coupled bile acid receptor, Gpbar1 (TGR5), is a membrane bound bile acid receptor. It is more broadly expressed, but its hepatic expression is limited to Kupffer cells, macrophages and sinusoidal endothelial cells. Little information regarding the effects of TGR5 in the pathological development of ALD has been reported before. In this study, we use C57BL/6(WT) and TGR5 knockout (TGR5−/−) mice to study the potential role of TGR5 in alcohol‐induced liver disease. Mice were challenged with chronic 5% ethanol feeding (10 days) plus a binge dose of alcohol by oral gavage (5 g/kg body weight) or with acute high dose alcohol gavage. The results showed that the liver/body weight ratio was increased in both WT and TGR5−/− ethanol fed mice, while it was significant higher in ethanol fed TGR5−/− mice than WT mice. The liver injury marker serum alanine transaminase (ALT) and aspartate aminotransferase (AST) were both increased. Alcohol feeding altered gene expression in bile acid homeostasis. The hepatic triglycerides were upregulated almost 30% higher in TGR5−/− mice than WT group. The haematoxylin Eosin (H&E) and Oil Red O staining showed more fat vacuoles in the liver of ethanol fed TGR5−/− mice than that in WT mice. We also treat the mice with a novel potent TGR5 agonist ligand and found protective effects on ALD in WT but not TGR5−/− mice. Collectively, these results suggest that TGR5 may prevent the development of ALD and represent a novel target for ALD treatment.

Support or Funding Information

City of Hope Departmental Internal funding
Teaching Difficult Physiological Concepts to the Medical Students Using Minicases, Educreations and Mentimeter
Authors: Ahmad Kashif Adil
Published: April 2020

Purpose

It is a consensus in medical schools that basic science courses is offered in a compressed curriculum, and medical students have difficulty in retaining essential basic science concepts. It is well established that medical, nursing, and healthcare schools use various pedagogical methods to deliver physiology curriculum. Our abstract focuses on medical education, it is always challenging for medical students to retain a plethora of physiology and pharmacology information. These concepts are considered high yield for passing standardized exams. We have shown the effectiveness of new modern tools in medical education that enhances the student learning experience.
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Methods

In collaboration with the students, we inquired and collected feedback and created a grid of physiological concepts that students find difficult to understand, examples include current multiplication in renal physiology, flow volume loops in respiratory.

We then planned three modes of content delivery methods. First, we designed short videos using novel technological tools like educreations clarifying those concepts. The students could view these videos at any time, anywhere, and on any device. We then offered mini‐cases in small groups that promoted higher‐level thinking on Bloom’s Taxonomy, for example, a case on diabetes insipidus that touched upon functions of ADH and integrated clinical reasoning. Finally, using Mentimeter (a novel audience response system), we conducted a short quiz as an interactive readiness experience for our students. The informal 15‐minute quiz was an effective way for students to gauge their performance. In this live session, the basic science instructor(s) were tasked to refresh, review, and validate physiology/pharmacology concepts with the students.

Results/Outcomes

According to our satisfaction surveys, students appreciated digital cognition. The survey response rate was 75% (n=24/32). More than 80% of students found the active learning sessions met their expectations or far exceeded their expectations. Examples of student narratives include, “The learning checkpoints and engaging activities with mini‐cases were good. It exceeded my expectations because I didn’t expect dedicated time to be given to that, but I am glad it did and would like to see similar things like this done in the future.” “The dedication of the instructor to providing learning materials in the forms of videos, readings, and mini‐lectures was a huge strength of the curriculum.”

Discussion

We conclude that our method of content delivery will provide an ideal opportunity for all students to learn the importance of contextual and difficult physiological concepts. The approach can help in identifying knowledge gaps and can serve a useful model for curricular delivery in medical schools, nursing, or any healthcare school that would like to promote critical thinking with a deeper understanding of physiology integrated with clinical science.