bluuebunny/crossref_metadata_2025 huggingface dataset:
P185 Patient perceptions of physical activity following a diagnosis of giant cell arteritis: a multinational qualitative study
Authors: Austin Keziah , Dures Emma , Almeida Celia , Cramp Fiona , Gilbert Kate , Guly Catherine , Hill Catherine , Mackie Sarah , O'Brien Anne , Watts Richard , Robson Joanna
Published: April 2020

Background

Giant cell arteritis (GCA) is the most common vasculitis in the UK, with an incidence of 220 cases per million in adults over 50 years of age. The physical symptoms as well as the side effects of glucocorticoids may impact patients’ ability to exercise. Maintaining physical activity (PA) has been shown to be beneficial to disease activity in other inflammatory conditions and is also a specific priority for GCA patients. The aim of this project was to explore patient perceptions of physical activity in GCA.

Methods

Multinational qualitative study, using interviews with 36 patients from the UK (25) and Australia (11), all of whom had a definitive diagnosis from imaging or biopsy. Interviews were recorded, transcribed, and analysed using inductive thematic analysis. This is secondary analysis of data collected to explore health-related quality of life in people with GCA.

Results

107 individual themes were reported by patients, which divided broadly into two overarching themes: barriers to (59) and facilitators of (48) physical activity. Four sub-themes were identified within each overarching theme. For barriers, these were: negative physical symptoms (including effects of disease flares, visual loss, fatigue, weakness, pain and stiffness), lack of physical capability (including poor stamina, confidence and mobility), negative perceptions around PA, and negative reinforcement (i.e. new physical symptoms following PA). Facilitators of physical activity were also grouped into four sub-themes: external facilitators (including motivation from healthcare professionals), access to appropriate facilities, personal strategies (including pacing and goal-setting) and personal facilitators (including internal motivation to improve symptoms, and positive reinforcement from noticing physical and psychological benefits of PA).

Conclusion

There are a variety of barriers to physical activity in GCA patients, including patients being fearful of exercise. In other inflammatory conditions, patients report improved symptoms following physical activity as well as wider benefits to general wellbeing and cardiovascular health. Education, motivational interviewing, and personalised strategies may be beneficial components of an intervention to support physical activity in patients with GCA.

Disclosures

K. Austin None. E. Dures Grants/research support; Has previously received an independent learning grant from Pfizer, however the work has been completed and the grant has been closed. C. Almeida None. F. Cramp None. K. Gilbert None. C. Guly None. C. Hill None. S. Mackie None. A. O'Brien None. R. Watts None. J. Robson None.
P194 Non-bacterial osteitis syndromes: characterising an adult population
Authors: Gill Tiara , Bubbear Judith , Keen Richard
Published: April 2020

Background

Inflammatory bone syndromes, such as SAPHO (synovitis, acne, pustulosis, hyperostosis and osteitis) and CRMO (chronic recurrent multifical osteomyelitis) are rare and have not been extensively described in the existing literature. Although there have been proposed classification and diagnostic criteria, there is still limited consensus and uniformity of approach in identifying and treating these patients. This may be due to a lack of data and understanding about the nature and prognosis of these conditions. Attempts to characterise, especially in adult populations, has been limited thus far. As a tertiary metabolic bone service with an allied regional sarcoma service, we present further data on these conditions in an adult population.

Methods

Retrospective analysis of notes of patients, with a diagnosis of SAPHO and CRMO, that were registered on a departmental database was performed. Data on demographics, symptom onset and character, serum and radiological tests and treatment were analysed.

Results

77 adults were identified with a diagnosis of SAPHO and CRMO, with a 60:40 % split respectively. Female preponderance was noted with 74% of this cohort represented. SAPHO patients tended to present slightly later with a mean age of symptom onset of 37 years as compared to 24 years in the CRMO group. Diagnostic delay was evident with mean time to diagnosis averaging 34 months. Pain and bony swelling were the most common symptoms (99% and 39% respectively), with the clavicle, sternum and spine being the most common sites involved. Osteitis was a universal feature in both groups, whereas skin and inflammatory joint involvement was more of a feature of SAPHO patients (59% vs 39%). There is limited consensus on appropriate diagnostic tests. This was reflected in this cohort with variable testing of inflammatory markers, serological testing of HLA B27 and extent and modality of imaging. Non-steroidal anti-inflammatories (NSAIDS), intravenous bisphosphonates, steroids and anti-microbials were all employed as treatment options, although efficacy was difficult to establish in this retrospective analysis, a significant source of referrals were from the local Sarcoma Unit; this may have introduced a degree of bias in our results.

Conclusion

As rare conditions, promoting awareness, having an index of suspicion and early referral to specialist centres would help address current issues faced by this patient population. Several national and international disease registries are in development and will undoubtly help to standardise data collection in order to guide further work in this area. Future studies examining treatment efficacy in current as well as emerging biological therapies, such as anti-tumour necrosis factor, anti- interleukin 1 and 17 agents, would be of significant value.

Disclosures

T. Gill None. J. Bubbear None. R. Keen None.
P196 Predictors of fracture in patients with primary hyperparathyroidism
Authors: Mariannis Demetris , Bukhari Marwan
Published: April 2020

Background

Primary hyperparathyroidism (PHPT) is a recognised risk factor for fractures as the persistently elevated parathyroid hormone levels are associated with lower bone mineral density (BMD). In early osteoporosis (<65 years), there is mainly trabecular bone loss while with increasing age, the largest loss of absolute bone mass occurs in cortical sites. Our aim was to compare cortical versus trabecular bone sites to determine which sites better predict fracture in PHPT patients referred for dual-energy X-ray absorptiometry (DEXA) scan in an observational cohort.

Methods

508 patients with PHPT referred for DEXA scan at Royal Lancaster Infirmary (UK) between 2006-2016 were included in a retrospective data analysis. Data collected included femoral and lumbar BMD, age, gender, height, weight, BMI, rheumatoid arthritis, smoking, alcohol, family history of fractures, secondary osteoporosis and long-term steroid use and was used to look at predictors of fractures using BMD at different sites. Simple statistics using independent t-tests and chi-squared tests were initially used and subsequently, logistic models were fitted unadjusted and adjusted confounding variables.

Results

97 males (19%) and 411 females (81%) were included, with 111 patients sustaining at least one fracture (21.9%). Mean age was 65 years (SD 0.55). Advanced age, female gender and a reduced BMD were all statistically significant factors associated with an increased risk of fractures. Smoking and long-term steroid use were also shown to increase risk of fracture however these were non-statistically significant. Adjusting for age and gender, a reduced L4 BMD was the strongest statistically significant predictor of fracture (OR 0.139, 95% CI: 0.112-0.782). Multivariate analysis revealed that a reduced L3 BMD was the strongest statistically significant predictor of fracture (OR 0.170, 95% CI: 0.071-0.72).

Conclusion

Overall, our study suggests that a reduced BMD at trabecular sites is a stronger predictor of fracture compared to cortical sites [Table 1]. Older age and female gender significantly increased the risk of fracture in PHPT patients. To the best of our knowledge, this is the first study looking at specific bone sites as independent predictors of fracture in PHPT patients. This analysis should be repeated using a larger cohort to validate our results.

Disclosures

D. Mariannis None. M. Bukhari None.
P202 Monitoring of immunoglobulin levels in RA patients on anti CD20, rituximab
Authors: Rice Ceara , Ghandi-Das Dhivya , Negi Anurag
Published: April 2020

Background

Rituximab is an anti-CD-20 used in resistant RA. It functions by reducing the efficacy of B cells which participate in this pro-inflammatory process. The BSR recommend immunoglobulin monitoring when using rituximab to aid rapid detection of hypogammaglobinaemia and address its potential complications which have been suggested by a recent large study in 2018 in the UK. Our objectives included: Are we measuring immunoglobulin (Ig) levels prior to rituximab infusions in all RA patients? Are we measuring Ig levels ‘4-6 months post infusions’ as per BSR guidelines? Is the cumulative dose of rituximab associated with hypogammaglobinaemia?

Methods

We collected data from departmental records for all the RA patients who were initiated on rituximab between 30/1/15 and 30/1/19. We recorded their basic demographics, pre-infusion immunoglobulin levels and subsequent levels with repeated infusions. All data was anonymised, but some patients had to be excluded from the final analysis due to missing immunoglobulin levels.

Results

42 patients were identified via the methodology. 88% of the 42 had Ig’s checked before starting rituximab. Only 38% of 37 patients included in this subsequent analysis had Ig’s checked within the recommended 6 months after their initial rituximab infusion. We also observed a notable average percentage drop in IgG levels after receiving approximately 6g of rituximab therapy, with 2 new patients developing new hypogammaglobinamia after 8g-10g of rituximab. After 10g of rituximab we noted an average percentage drop in rituximab of 26.8%. However, no statistical correlation between hypogammaglobinaemia and infection rates was found.

Conclusion

Although no correlation between hypogammaglobinaemia and infection rates were found, this study used small numbers from a single centre only. To improve UHW’s monitoring of Ig’s we introduced a performer which aims to help recording of pre and post IgG levels and recording of infections experienced between infusions. We hope this will increase identification of newly developed hypogammaglobinaemia and increase awareness of infection rates, ultimately leading to an improvement in patient care.

Disclosures

C. Rice None. D. Ghandi-Das None. A. Negi None.
P228 Risk of sinusitis in patients with rheumatoid arthritis: association with different treatment strategies
Authors: Halai Kapil , Bechman Katie , Norton Sam , Cope Andrew P , Hyrich Kimme L , Galloway James B
Published: April 2020

Background

There is an established increase risk of infection in rheumatoid arthritis (RA). Most published data examine serious events, which affect 3-5% of patients annually. Approximately half of the patients experience non-serious infection, which does not require parental antibiotics or hospitalisation. Sinusitis events affecting patients on biologics (particularly TNF inhibition) have been observed but not well described, with a prevalence of 2-17%. Sinusitis is associated with treatment discontinuation and surgical interventions. We aim to describe the rates of sinusitis with different RA treatment strategies and provide estimates of risk difference.

Methods

The population was adults with RA, enrolled in the British Society for Rheumatology Biologics Register for Rheumatoid Arthritis registry (BSRBR-RA) during the first 3 years of study on biologic treatment. The predictor was treatment strategy at the time of event (csDMARD only, TNF inhibitors, IL-6 inhibitors and anti-CD20). The primary outcome was sinusitis according to MeddraPT codes reported by patients (via patient diary) or clinician (on an adverse event form). A multi-failure Cox survival model was used to compare risk across treatment groups. A multivariate model adjusted for confounders (age, gender, entry year in BSRBR-RA, disease duration, baseline disease activity by DAS-28, HAQ, comorbidities, smoking status and steroid use.

Results

23,584 patients were included, contributing to a total of 64,035 patient-years of follow up. There were 797 infective events in 580 patients. This corresponded to a rate of 0.95 per year (95% CI 0.87 to 0.1.02). The sinusitis rate was numerically highest with anti-CD20 (Rituximab) and lowest with csDMARDs (Table 1). In the adjusted Cox proportional hazard model, all 3 biologics strategies had a significantly higher rate of sinusitis compared to csDMARD cohort; (TNFi 2.76 (95%CI 1.85 to 4.12); IL-6R 2.51 (1.00 to 6.29); anti-CD20 3.14 (1.46 to 6.72).

Conclusion

Sinusitis is more common in patients prescribed biologics compared to the csDMARD. The highest rate was seen with rituximab. This would fit biologically, as the primary immunodeficiency phenotype of recurrent sinusitis aligns with antibody defects e.g. common variable immunodeficiency. However, it is interesting to observe that sinusitis is more common across the treatment classes.

Disclosures

K. Halai None. K. Bechman None. S. Norton None. A.P. Cope None. K.L. Hyrich Honoraria; AbbVie paid to the institution and grant income from Pfizer and Bristol-Myers Squibb for activities outside of this work. J.B. Galloway Honoraria; or speaking or attending conferences from AbbVie, Bristol-Myers Squibb, Celgene, Janssen, Pfizer and Union Chimique Belge.
P274 Association of self-reported symptoms and behaviour in axSpA: initial analyses from the Project Nightingale study
Authors: Barnett Rosie , Ng Stanley , Jones Simon L , Young Matthew , Sengupta Raj
Published: April 2020

Background

Axial spondyloarthritis (axSpA) is a chronic inflammatory disease primarily affecting the spine and sacroiliac joints, characterised by fluctuating periods of flare and remission. Management is based on patient-reported symptoms and outcome measures collected at follow-up appointments which may be subject to recall bias. Smartphone technologies for monitoring disease symptoms provide an opportunity to gain a more complete understanding of disease burden and symptom patterns and may facilitate optimisation and personalisation of axSpA management.

Methods

Patients with axSpA attending the Royal National Hospital for Rheumatic Diseases in Bath were invited to participate in the study. Through the uMotif symptom tracking app, patients were sent daily reminders to log pain, fatigue, sleep, recommended exercise, mood and stress using 5-point Likert scales (higher scores indicating better outcomes eg. No pain=5), in addition to optional variables such as screen time, caffeine intake and menstrual cycle. For each patient, at each time-point, Spearman rank correlation coefficients were used to evaluate inter-variable correlations. Lagged variables were used to assess correlations with variables from the previous day (variable-1).

Results

Between 05/04/18 and 08/03/19, 174 patients consented for research and logged a mean of 99.73 (SD = 99.97) days of data (range=1 - 323 days). Significant correlations were identified between uMotif variables (Table 1), including pain and fatigue; mood and stress; hot flushes and fatigue/pain; confidence in self-management and mood/pain; smoking and pain/fatigue; exercise and sleep; exercise and mood - supporting existing evidence regarding exercise implementation in axSpA.

Conclusion

These findings demonstrate relationships between a variety of patient-reported symptoms, including variables that to our knowledge, have not yet been explored in axSpA. In future research, it will be important to determine whether there is a chronological pattern within an individual or combination of variables that could predict a flare. Greater understanding of axSpA disease patterns and identification of the optimal timing of intervention to ameliorate these symptoms may ultimately reduce flare frequency, duration and intensity, and greatly improve quality of life for patients with axSpA.

Disclosures

R. Barnett None. S. Ng None. S.L. Jones None. M. Young None. R. Sengupta None.
Semantic Search in Psychosis: Modeling Local Exploitation and Global Exploration
Authors: Lundin Nancy B , Todd Peter M , Jones Michael N , Avery Johnathan E , O’Donnell Brian F , Hetrick William P
Published: April 2020

Impairments in category verbal fluency task (VFT) performance have been widely documented in psychosis. These deficits may be due to disturbed “cognitive foraging” in semantic space, in terms of altered salience of cues that influence individuals to search locally within a subcategory of semantically related responses (“clustering”) or globally between subcategories (“switching”). To test this, we conducted a study in which individuals with schizophrenia (n = 21), schizotypal personality traits (n = 25), and healthy controls (n = 40) performed VFT with “animals” as the category. Distributional semantic model Word2Vec computed cosine-based similarities between words according to their statistical usage in a large text corpus. We then applied a validated foraging-based search model to these similarity values to obtain salience indices of frequency-based global search cues and similarity-based local cues. Analyses examined whether diagnosis predicted VFT performance, search strategies, cue salience, and the time taken to switch between vs search within clusters. Compared to control and schizotypal groups, individuals with schizophrenia produced fewer words, switched less, and exhibited higher global cue salience, indicating a selection of more common words when switching to new clusters. Global cue salience negatively associated with vocabulary ability in controls and processing speed in schizophrenia. Lastly, individuals with schizophrenia took a similar amount of time to switch to new clusters compared to control and schizotypal groups but took longer to transition between words within clusters. Findings of altered local exploitation and global exploration through semantic memory provide preliminary evidence of aberrant cognitive foraging in schizophrenia.
INHIBITION OF FERROPTOSIS BY LIPROXSTATIN‐1 COMPOUND PROTECTS THE MYOCARDIUM AGAINST ISCHEMIA/REPERFUSION INJURY BY DECREASING VDAC1 LEVELS AND INCREASING IN GPX4 ACTIVITY
Authors: Feng Yansheng C. , Imam Aliagan Abdulhafiz C. , Tombo Nathalie C. , Bopassa Jean C.
Published: April 2020

Ferroptosis is a non‐apoptotic form of cell death characterized by iron‐dependent accumulation of lethal lipid reactive oxygen species (ROS) and liproxstatin‐1 is a potent inhibitor of ferroptosis. We investigated whether Liproxstatin‐1 can protect heart against ischemia/reperfusion (I/R) injury and determine the mechanism underlying cardioprotection. Isolated C57BL/6J mice hearts were subjected to 35 min ischemia followed 120 min reperfusion and treated with Liproxstatin‐1 (200nM). Myocardial infarct size was assessed at the end of the reperfusion, mitochondrial integrity was measured by analyzing cristae morphology and ROS production was assessed using amplex red method. The calcium retention capacity (CRC) required to induce mitochondrial permeability transition pore (mPTP) opening was measured using calcium green dye. The levels of mitochondrial proteins cyclophilin D (CypD), Glutathione peroxidase 4 (GPX4), VDAC1, 2 and 3 were assessed by Western blot; and GPX4 activity was measured using a kit. We found that
liproxstatin‐1 treatment decreased myocardial infarction size, ROS production, and VDAC1 expression as well as preserved mitochondrial structural integrity by protecting cristae from damage. These mitochondrial effects of
liproxstatin‐1 were associated with the increase in the GPX4 levels and activity. But,
liproxstatin‐1 did not affect the CypD, VDAC2, and VDAC3 expression as well as mitochondrial CRC. Together, these result indicate that
liproxstatin‐1 treatment protects myocardium against I/R injury by increasing GPX4 activity resulting in the decrease in mitochondrial ROS production; preserving mitochondrial structure as well as decrease in VDAC1 (but not VDAC 2 and 3) expression. Liproxstain‐1 cardioprotective action does not involve the regulation of the mPTP opening. These results point to the cardioprotective role of anti‐ferroptosis compound after I/R.

Support or Funding Information

[NIH RO1 grant HL138093 (JCB)]
Functional Characterization of Position 271 in NTCP, a Predicted Rheostat Location
Authors: Ruggiero Melissa , Malhotra Shipra , Karanicolas John , Swint-Kruse Liskin , Hagenbuch Bruno
Published: April 2020

Background

Human Na+/taurocholate cotransporting polypeptide (NTCP) is a sodium dependent bile acid transporter expressed at the basolateral membrane of human hepatocytes. It is important for the enterohepatic circulation of bile acids, such as taurocholate (TCA), but it also transports other compounds including estrone‐3‐sulfate (E3S) and rosuvastatin. We previously identified a rheostat position at a naturally occurring polymorphism, S267F. That is, when other amino acids were substituted into this position, progressive changes in function were observed.

Aim

The purpose of the current study is to determine whether position N271, a predicted rheostat, is indeed another rheostat position in NTCP.

Methods

The crystal structures of two bacterial NTCP homologs were used to model the structure of human NTCP in the inward‐open and outward‐open conformations. Mutations were then introduced into the structural models and the energetic effect on protein stability was evaluated using the Rosetta software. Based on these calculations we predicted that N271 is a rheostat. Mutants for position N271 were generated using site‐directed mutagenesis and the resulting proteins were transiently expressed in HEK293 cells. The functions of the mutants were characterized by measuring uptake of radiolabeled model substrates: [3H]‐taurocholate, [3H]‐estrone‐3‐sulfate, and [3H]‐rosuvastatin. Plasma membrane expression levels were determined by surface biotinylation assays.

Results

Initial transport studies using nanomolar concentrations of TCA, E3S, and rosuvastatin indicated a variety of intermediate functional outcomes for the mutants. Furthermore, this alteration in function was substrate dependent, which is consistent with our findings for S267. Some mutants, such as N271A, transport substrates similar to wild‐type NTCP, while others, like N271K, show variation depending on the substrate. N271P consistently transports all substrates minimally. In addition, there is no correlation between the initial uptake results and the Rosetta energies.

Summary and Conclusions

Initial transport studies demonstrate that position N271 is a rheostat position, as predicted by the alteration in Rosetta energy scores. However, there was no correlation between the calculated energy scores and the functional results. This suggests that functional outcomes of specific amino acid replacements at position N271 may arise from more complex conformational changes than were captured by the two modeled states. Additional experiments should help to determine to what extent the observed change in function is due to variations in the substrate affinity, the turnover number or the transport capacity of the mutants.

Support or Funding Information

P30 GM118247, GM077336, and W. M. Keck Foundation
Sprouting Angiogenesis Contributes to Coronary Collateral Growth Induced by Repetitive Ischemia in Adult Mice
Authors: Jamaiyar Anurag , Juguilon Cody , Richardson Devan , Gadd James , Wang Tao , Enrick Molly , Goodner Rayana , Chilian William M. , Yin Liya
Published: April 2020

In the United States, coronary heart diseases (CHD) are the leading cause of mortality and morbidity. A well‐developed coronary collateral circulation ameliorates the consequences of CHD, reducing the incidence of sudden death and infarct sizes following coronary occlusion. Stimulation of coronary collateral growth (CCG) is also an alternative therapeutic approach to patients with intractable angina pectoris. The importance of CCG is undisputable, but the process and mechanism underlying CCG is unclear. We developed a mouse model of CCG induced by repetitive ischemia (RI) and validated CCG by contrast echocardiography to measure the coronary blood flow in the normal zone (NZ) and the collateral dependent zone (CZ). We also used Micro‐CT scans to reconstruct the coronary vasculature in 3D and quantify CCG. In this study, we crossed ROSA mT/mG double fluorescent reporter mice with cell type‐specific cre mice to investigate the roles that different cell‐types play at various stages of CCG. Mice were subjected to the RI protocol and their cardiac function and coronary blood flow was measured by echocardiography. Following sacrifice, hearts were fixed for immunostaining or imaging under confocal and multiphoton microscopy. In ROSA mT/mG floxed, Apln cre mouse hearts, large vessels (diameter of 20–50 μm) expressing GFP were identified in the collateral dependent zone in the early stage of RI, indicating that they originated from sprouting endothelial cells. At the end of RI, a complete network of these vessels was observed anastomosing with existing vasculature from the NZ. Immunostaining revealed an outer layer of smooth muscle cells in these vessels, suggesting the arteriogenesis or arterialization. Interestingly, in ROSA mT/mG floxed, Lyz2 cre mouse hearts, GFP expressing myeloid cells were observed in high numbers around newly formed large vessels, suggesting that immune cell recruitment occurs in the early stages of CCG. Our preliminary data show that coronary collaterals form in response to RI, are of microvascular origin and are associated with early recruitment of immune cells to the CZ. Our further study will focus on the process of CCG in greater detail at the molecular and cellular level, potentially leading toward a therapeutic application of induced CCG in patients of ischemic heart diseases.

Support or Funding Information

The research is funded by National Institutes of Health grant 2R01HL103227‐05 (YZ, LY), 1R01HL135110‐01 (WMC, LY), 1R01 HL137008‐01A1 (LY) from American Heart Association (LY).
Mitochondrial Thioesterases: Localization, Regulation and Functional Relevance of Acyl‐CoA Thioesterase 2
Authors: Bekeova Carmen , Anderson-Pullinger Lauren , Boye Kevin , Sharpadskaya Yana , Seifert Erin
Published: April 2020

Acyl‐CoA thioesterases (Acots) hydrolyze fatty acyl‐CoA esters into free fatty acid and CoA. This group of understudied enzymes is found in several cellular compartments including mitochondria where Acots have been hypothesized, but never shown, to prevent a CoA limitation and mitigate beta‐oxidation overload. Before embarking on the study of the functional role of mitochondrial Acots we sought to determine some basic characteristics of mitochondrial Acots (Acot2, 7, 9 13) that were unknown but would be important for the interpretation of physiology in models with Acot depletion. First we determined that Acot7, 9 and 13 localize to the mitochondrial matrix, joining Acot2 that was already shown to localize there. Then, we determined that Acot13 activity is powerfully inhibited by CoA, as was previously shown for Acot7 and Acot9, whereas Acot2 activity is unaffected by CoA, and was not substantially altered by any of the other potential allosteric regulators we tested. Thus Acot2 may be the only matrix Acot that is constitutively active and, importantly, remains fully active when CoA is plentiful. Finally, using measurements of protein abundance and activity in isolated mitochondria from multiple mouse tissues, we found that the highest thioesterase activity is towards short chain acyl‐CoA in brown adipose tissue, kidney and skeletal muscle, mediated by Acot9; however CoA inhibition would largely suppress this activity when CoA is plentiful. The next most significant thioesterase activity was towards C14:0‐ and C16:0‐CoA, in heart and skeletal muscle, and we found that this activity was shared equally between Acot2 and Acot13. However, as for Acot9, Acot13 activity would be suppressed when CoA is plentiful, leaving Acot2 as the sole source of long‐chain acyl‐CoA thioesterase activity in heart and skeletal muscle when CoA is replete, suggesting that it should have functional relevance in these tissues. Indeed, using Acot2 whole‐body and muscle‐specific knockout mouse models, we find that cardiac ejection fraction is decreased when Acot2 is depleted, but only in female mice. This was accompanied by higher cardiac acyl‐CoA levels, across almost all chain lengths, in female mice with Acot2 depletion whereas Acot2 depleted male hearts had acyl‐CoA levels that were similar to control mice. Substrate oxidation studies in isolated heart mitochondria reveal differences with fatty acid substrates but not with pyruvate, suggesting that beta‐oxidation is the major substrate pathway affected by Acot2 deletion. Ongoing studies aim to further understand the functional relevance of Acot2 in the heart. Also, because quantitative analyses suggest that Acot2 activity could theoretically outstrip acyl‐CoA dehydrogenase activity which has a similar Kd for long‐chain acyl‐CoAs, we are interested to understand where, within the matrix, Acot2 localizes and, in this regard, are investigating the binding partners of Acot2.
Correlation of Stroop Effect in Medical Students to Their Performance in Pharmacology National Board of Medical Examiners (NBME) Examination
Authors: Panta Raju , Paudel Keshab Raj , Toth Stephen , Olsen John
Published: April 2020

Objective

The Stroop test is used to assess selective attention in cognitive psychology. Interference between word meaning and ink color in Incongruent Stroop test prolongs reaction time, which involves the brain searching for some collection of the stored information for prompt use. This is called the Stroop effect. Previous studies have reported the strong correlations between the Stroop effect and academic performance. Therefore, we aimed to study the correlation of the Stroop effect in medical students to their performance in pharmacology NBME examination.

Methods

For this study, 40 medical students who completed their pharmacology NBME examination were selected. The Stroop, congruent and incongruent, tests were performed using the EncephalApp Stroop Test (Android/iOS). In Congruent test (Stroop Off), the subjects were asked to select ink color of the hashtag (#) symbols printed in different ink colors (red, blue, and green). In Incongruent test (Stroop On), the subjects were asked to select the ink color of the words RED, BLUE, and GREEN printed in different ink colors, e.g., the word “RED” printed in blue ink had to be selected as blue. Each subject practiced each test for two attempts and 5 attempts of each test were then recorded.

The EncephalApp Stroop Test automatically recorded the results of the tests as duration in seconds, which include Congruent Stroop (OffTime duration), Incongruent Stroop (OnTime duration), OffTime plus OnTime duration, OnTime minus OffTime duration. The results of OnTime minus OffTime duration of the Stroop test and pharmacology NBME examination scores were tabulated in Microsoft Excel and descriptive analysis was done. The correlation of the “OnTime minus OffTime duration” to pharmacology NBME examination scores was determined by a Pearson correlation coefficient and regression analysis.

Results

The correlation coefficient (adjusted r2) of our study was −0.0255 and significance F was 0.862. The “OnTime minus OffTime duration” in medical students was not correlated with the pharmacology NBME examination score (r (38) = 0.0283, p = 0.862). The regression model predicted only 0.08% (R2 = 0.0008) of the pharmacology NBME examination score of medical students.

Conclusion

The Stroop effect in medical students is insignificantly correlated with their performance in pharmacology NBME examination. However, correlations of the Stroop effect in medical students with their performance in other subjects and examinations need to be assessed in further studies.

Support or Funding Information

The study was supported by the Trinity Medical Sciences University School of Medicine, St. Vincent and the Grenadines.
Regulation of Coronary Collateral Growth by MicroRNA‐21 in Metabolic Syndrome
Authors: Juguilon Cody , Richardson Devan , Gadd James , Enrick Molly , Jamaiyar Anurag , Xu Yanyong , Wang Zhiyuan , Wang Tao , Kolz Chris , Chilian William , Yin Liya
Published: April 2020

Well‐developed coronary collaterals prove to be highly beneficial in salvaging ischemic myocardium, preserving cardiac function, and improving patient outcome post‐occlusion. However, this process of coronary collateral growth (CCG) is impaired in patients with metabolic syndrome. A complete understanding of the underlying mechanism, cell types, and genes contributing towards this impairment have yet to be elucidated. Therefore, uncovering more about the process may lead to potential therapeutics to induce CCG in metabolic syndrome. MicroRNA‐21 (miR‐21) is abundantly expressed in vascular and immune cells with numerous implications in cardiovascular disease including atherosclerosis, heart failure, and myocardial infarction. Furthermore, miR‐21 has been shown to regulate processes such as apoptosis, immune cell polarization, and endothelial progenitor proliferation. Additionally, miR‐21 dysregulation has been rooted in the diabetic population where lies a systemic inflammatory state. In this study, we investigated the role of miR‐21 in a mouse model of CCG.

Our preliminary data suggested that down‐regulating miR‐21 rescues impaired CCG in a diet‐induced model of metabolic syndrome. Thus, we investigated the underlying mechanism and focused on the roles of miR‐21 in the maintenance of vascular homeostasis, function and inflammatory responses in metabolic syndrome. First, we studied whether miR‐21 regulates endothelial homeostasis by modulating the function and homing of bone marrow stem cells in metabolic syndrome. We analyzed endothelial progenitor cells (CD34+) in bone marrow and peripheral blood, along with endothelial proliferation in WT, miR‐21 knockout, and metabolic syndrome mice. Second, we studied whether miR‐21 regulates the inflammatory response in metabolic syndrome by characterizing bone marrow derived macrophages from WT and miR‐21 knockout animals along with mapping their influence on vascular cells. Third, we utilized a myeloid specific miR‐21 knockout to study its role during CCG in vivo. We found CCG to be blunted in these animals which suggests the importance of myeloid derived miR‐21 in healthy mice. Further studies will give us more insight on miR‐21 and its regulation of CCG in metabolic syndrome.

Support or Funding Information

The research is funded by National Institutes of Health grant 2R01HL103227‐05 (YZ, LY), 1R01HL135110‐01 (WMC, LY), 1R01 HL137008‐01A1 (LY)from American Heart Association (LY).
Development of G Protein‐Coupled Estrogen Receptor (GPER) Ligands for Improved Efficacy and Aqueous Solubility
Authors: Green Michael , DeLeon Chelsea , Gunn Joseph , Duncan Emily , Olson Onika , Skibinski Karissa , Arnatt Christopher
Published: April 2020

The G‐protein estrogen receptor (GPER) has been implicated in a variety of disease states and conditions. Previously our group identified a novel GPER antagonist, CIMBA, that showed improved potency as compared to the G‐series antagonist, G‐36, and exhibited the ability to reduce the formation of gallstone in a murine model. Additional modifications were made to the CIMBA scaffold to improve potency and enhance solubility. In this study, particular emphasis was placed on examining the replacement of the cyclohexyl moiety in CIMBA. Groups explored included various cyclic aliphatic systems, aliphatic chains, conjugated ring systems, piperazine, piperidine, and morpholine. Additional modifications explored the tolerance of the additional groups at the methoxy of CIMBA as well as linker modifications. Compounds were initially screened for calcium mobilization at 10 mM. All compounds exhibiting either agonism or antagonism at 10 mM were screened further to determine the appropriate EC50 or IC50 values. In this series of compounds, it was determined that the absence of the amine in the linker was detrimental to activity. Additionally, the replacement of the cyclohexyl ring in CIMBA with a piperazine (referred to as PIMBA), showed improved aqueous solubility above 10 mM without effecting the potency drastically. Off‐target binding to the nuclear estrogen receptors, ERa and ERb, was determined with a fluorescence polarization. The improved solubility of PIMBA may increase the success of in vivo studies utilizing GPER‐specific ligands in understanding the pharmacology of the receptor in different disease states.
Young women are protected against leg endothelial dysfunction induced by adoption of a Westernized lifestyle
Authors: Smith James S. , Sales Allan K. , Walsh Lauren K. , Soares Rogerio Nogueira , Woodford Makenzie , Padilla Jaume , Manrique-Acevedo Camila
Published: April 2020

Background

A Westernized lifestyle is characterized by sedentary behavior and consumption of diets high in refined sugars. This lifestyle is associated with weight gain, insulin resistance and the development of cardiovascular disease (CVD). Endothelial dysfunction is one of the key events in the pathogenesis of CVD. We have previously demonstrated that short‐term exposure to physical inactivity results in popliteal artery endothelial dysfunction evaluated by flow‐mediated dilation (FMD) in healthy men. However, it is unknown if transition to a Westernized lifestyle differentially impacts upper‐ and lower‐limb vascular health of healthy young women versus men.

Methods

Twelve recreationally active women (aged 23±1 yrs; BMI 22.2±1.02 kg/m2) and 9 men (aged 27±2 years; BMI 24.73±1.27 kg/m2) were recruited from the community to undergo a 10‐day intervention period of sedentary behavior combined with consumption of six cans of high‐fructose carbonated soda per day. Ambulation, body composition, brachial (arm) and popliteal (leg) artery FMD and biochemical parameters were assessed at baseline and at the end of the intervention.

Results

Adoption of a sedentary behavior (reduction in ambulation from 12383±382 steps/day to 3992±151steps/day; p=0.0001) and high‐fructose consumption for 10 days did not result in significant changes in body weight or percent fat mass, but increased fasting triglycerides (basal: 63±7.26 vs. post: 79±9.23, p=0.04) and HOMA‐insulin resistance index (basal: 0.6±0.1 vs. post: 1.0±0.1, p=0.001), independent of sex. Notably, the Westernized lifestyle reduced popliteal artery FMD in men (basal: 5.7±1.0 vs. post: 3.1±0.8, p=0.04) but not in women (basal: 3.6±0.7 vs. post: 5.7±0.9, p=0.05). Brachial artery FMD remained unchanged following the intervention in both men and women.

Conclusions

Our results indicate that physically active young females, but not males, are protected from leg endothelial dysfunction induced by short‐term exposure to a Westernized lifestyle.

Support or Funding Information

none
Ventromedial Hypothalamic Nucleus (VMN) Neuroestradiol: Sex Mono‐Versus Dimorphic Control of Glycogen Metabolism and Glucoregulatory Transmitter Signaling
Authors: Uddin Md. Main , Ibrahim Mostafa M.H. , Briski Karen P.
Published: April 2020

The VMN functions within the neural glucostatic network. Estradiol exerts sex‐specific control of VMN nitric oxide (NO) and γ‐aminobutyric acid (GABA) transmission, which respectively stimulates or suppresses glucose counter‐regulatory hormone secretion. In the brain, the VMN is a principal site of neuroestradiol production by aromatase. This project utilized the aromatase inhibitor letrozole (LZ) alongside combinatory high‐resolution microdissection/Western blotting to address the premise that neuroestradiol governance of VMN metabolic sensory neuron protein marker [neuronal nitric oxide synthase (nNOS); glutamate decarboxylase65/67 (GAD)] expression is sex‐dimorphic. Glycogen metabolism impacts VMN nNOS and GAD profiles; here, LZ treatment effects on VMN glycogen synthase (GS) and phosphorylase brain‐ (GPBB; glucoprivic‐sensitive) and muscle (GPMM; norepinephrine‐sensitive) variant proteins were examined. VMN aromatase protein content was equivalent between the sexes; intracerebroventricular LZ infusion of testes‐intact male and ovariectomized, estradiol‐replaced female rats blocked insulin (INS)‐induced hypoglycemia up‐regulation of this profile. LZ elicited sex‐contingent changes in basal VMN nNOS and GAD expression, but blocked INS‐induced NO stimulation and GAD suppression in each sex. INS caused LZ‐reversible GPBB augmentation or suppression in male versus female, respectively. Male and female VMN GPMM content was correspondingly unchanged or decreased by INS; LZ reduced baseline GPMM expression in females only, but did not attenuate INS regulation in either sex. LZ correspondingly down‐ or up‐regulated baseline pyruvate recycling pathway marker protein expression in males (glutaminase) and females (malic enzyme‐1), and altered INS effects on the same proteins. Results show that neuroestradiol is required in each sex for optimal VMN metabolic transmitter signaling of hypoglycemia‐associated energy deficiency. Locally‐generated estradiol steroid involvement in sex‐specific hypoglycemic patterns of VMN glucoprivic‐sensitive GPBB expression may correlate with sex‐dimorphic glycogen mobilization during this metabolic stress. Neuroestradiol may also exert sex‐specific effects on glucogenic amino acid energy yield by actions on distinctive enzyme targets in each sex.

Support or Funding Information

National Institutes of Health DK‐109382
Souchinone Inhibits Mesangial Cell Fibrosis and Inflammatory Response Through TGF‐β1/Smad signaling and NLRP3 inflammasome pathway
Authors: Yoon Jung Joo , Han Byung Hyuk , Lee Hyeon Kyoung , Lee Yun Jung , Ahn You Mee , Kang Dae Gill , Lee Ho Sub
Published: April 2020

Mesangial proliferative glomerulonephritis is characterized by an increased number of mesangial cells in the glomeruli in the kidneys and damage to the glomeruli by activation of some growth factors (e.g., angiotensin II) and deposition of extracellular matrix proteins. However, the effect of sauchinone on mesangial proliferative glomerulonephritis is still unclear. In the present study, we investigated the protective effect of sauchinone against angiotensin II (AngII)‐induced proliferation and fibrosis in human renal mesangial cells. Thus, this study was investigated the inhibitory effect of sauchinone (0.1–1 μM) on AngII‐stimulated mesangial cells proliferation and fibrosis. Sauchinone, one of the active lignan isolated from Saururus chinensis, was reported to possess broad bioactivities, such as anti‐inflammatory, anti‐oxidant, anti‐tumor, anti‐apoptosis and anti‐obesity effects.

In this study, thymidine incorporation under AngII was significantly accelerated, which was inhibited by sauchinone in a dose dependent manner. In migration assays, a significant reduction in wound surface was observed in pretreatment of sauchinone. Sauchinone induced down‐regulation of cyclins/CDKs and up‐regulation of CDK inhibitor, p21waf1/cip1 and p27kip1 expression. In addition, AngII enhanced expression of fibrosis biomarkers such as collagen IV and CTGF, which was markedly attenuated by sauchinone. Sauchinone decreased TGF‐β1 and Smad‐2/Smad‐4 expression, whereas increased Smad‐7 expression under AngII. Furthermore, sauchinone inhibited AngII‐induced inflammatory factors level as well as NF‐κB p65 translocation and ROS production.

These results suggested that sauchinone may alleviate mesangial proliferation and inflammation possibly involved in renal fibrotic process, further mesangial proliferative glomerulonephritis through disturbing TGF‐β1/Smad signaling and NLRP3 inflammasome pathway. Thus, sauchinone might prove to be effective in the treatment of renal dysfunction leading to mesangial proliferative glomerulonephritis.

Support or Funding Information

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIP) (2017R1A5A2015805)(2018R1D1A1B07050720).
SIRT4 Regulates Lysine Malonylation in Ovarian Granulosa Cells
Authors: Min Zheying , Xing Zeyu , Yu Yang
Published: April 2020

SIRT4 belongs to the sirtuin family, which comprises NAD+ dependent lysine decaylases. SIRT4 has ADP‐ribosyltransferase, lipoamidase and decaylase activeties. In the study, we observed differential regulated lysine malonylationin ovarian granulosa cell lines (KGN) by the lable‐free quantitive technique. In SIRT4 depleted cells, there were a total of 65 malonylationsites from 56 proteins, including 39 up‐regulated malonylated proteins and 17 down‐regulated proteins. The differential regulated malonylation proteins were enriched in metabolic process and response to stimulus in GO biological process. We also detected the malonylation levels of PGK1 K131 and ENO1 K80 sites were decreased significantly in SIRT4 depleted cell lines, which were the same sites of PGK1 and ENO1 changed in PCOS primary ovarian granulosa cells (GCs). In addition, SIRT4 was decreased in PCOS GCs. In a conclusion, SIRT4 may regulate mitochondrial metabolic ability in PCOS GCs through modifying lysine malonylation of proteins in glycolysis pathway.

Support or Funding Information

This work was supported in part by the National Key R&D Program of China (2017YFC1001003, 2018YFC1003203), the National Natural Science Funds for general program (81571400, 81771580).
Antibacterial Screening Of Ethanolic Pomegranate Peel Extract
Authors: Jackson Destiny , Abu-Niaaj Lubna Fawzi
Published: April 2020

Pomegranate (Punica granatum L.) is reputed for its nutritional properties. This fruit is rich in antioxidants, vitamins, and minerals. The juice industry produces a high quantity of peel that is thrown away despite of its richness of bioactive compounds. The increased demand for environmentally friendly pharmaceutical agents brought attention to commercializing the use of fruit peel.

Objectives

This study aims to 1) evaluate the antibacterial activity of ethanolic pomegranate peel extract on six bacteria that cause food spoilage or are associated with human diseases., and to 2) determine the effect of temperature on the antimicrobial activity induced by the pomegranate peel ethanolic extract.

Material and Methods

The growth inhibition by the pomegranate peel extract was evaluated on Bacillus subtilis, Bacillus megaterium, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Micrococcus luteus. Fresh pomegranate peel was dried, grinded into fine particles, and stirred in ethanol for 30 minutes either at room temperature or with gentle boiling. Both extract preparations were filtered, wrapped with aluminum foil to avoid photooxidation. The pH measurement was recorded immediately, then daily for at least 30 days. The crude (gummy) material is a concentrated extract which was tested on bacteria at a concentration of 40ug/μl. The antibacterial activity of was determined using agar well diffusion method. The bacterial were cultured into prepared nutrients broth and incubated at 37°C for 24 h and standardized to 0.5 Mc‐Farland Scale (108 cfu/mL) in a prepared normal saline. The cell suspensions were seeded into prepared nutrients agar plates, and wells of 8mm were bored into the agar. The screening was done by placing 100ul in a well of either the extract, or the crude that was prepared at 40ug/μl.

Results

The ethanolic extract and crude of pomegranate peel showed varying degree of antibacterial activity against the tested bacteria. The inhibition zone range was between 14–28 mm and it was sustained for up to 72hr. The high temperature preparations showed a minimum of 10% more antibacterial activity. The pH measurements showed stability for at least 30 days for all preparations kept in the refrigerator.

Conclusion

It appears that the pomegranate peel has the potential for use in food applications as a natural preservative, and also in pharmaceutical applications.

Support or Funding Information

NSF‐IPSRG, LSAMP Grant and Land Grant