bluuebunny/crossref_metadata_2025 huggingface dataset:
Hematological status of newly‐calved cows with mineral metabolism disturbance
Authors: Plemyashov Kirill , Nikitin Georgiy , Nikitin Anastasiya , Kovalev Sergej , Trushkin Vyacheslav , Anipchenko Polina , Votinceva Aleksandra , Batrakov Aleksej
Published: June 2021

In many livestock farms in Russia there is a problem of correct balancing rations by the major mineral elements. Mineral metabolism disturbance are accompanied by various pathologies from all body systems, including the reproduction function. Timely conducting clinical examination, including hematological examination of all groups of animals allows to prevent the development of negative conditions in the body of cows associated with impaired mineral metabolism. During 2018, 74 blood samples obtained from newly‐calved cows were investigated. All the animals had various signs of mineral failure ‐ lordosis of the lumbar spine, osteomalacia of the caudal vertebrae and the last pair of ribs, weakness of the ligaments of the finger joints. With biochemical research of serum determined that the concentration of total protein averaged 90.7 ± 5.9 g/l, albumin −27.3 ± 3.7 g/l, globulin −61.4 ± 3.2 g/l, which indicates hypoproteinemia with dysproteinemia, with the concentration urea increased to 9.5 ± 0.4 mmol/l. The alkaline phosphatase level in fresh cows was 122.1 ± 11.1 IU/l, AST activity was 130.8 ± 5.6 IU/l, and ALT −34.7 ± 3.9 IU/l. The concentration of cholesterol and triglycerides in the blood of the studied animals were quite high and amounted to 8.7 ± 0.3 mmol/l and 0.29 ± 0.06 mmol/l, accordingly, indicating a violation of lipid metabolism. Ca : P ratio in the cows' blood was disturbed and was often 2: 1.9. The amount of total calcium in the blood was at the level of 2.25 ± 0.03 mmol/l, and of phosphorus −2.09 ± 0.03 mmol/l. During the morphological research, it was determined that the number of red blood cells in cows was lower than the reference values and amounted to 4.3 ± 0.4 * 10^6 /μL, the hemoglobin concentration was 84.0 ± 4.8 g /l, anemia was observed in erythropenia and hypochromemia. Also, in cows with signs of impaired mineral metabolism in the postpartum period, a decrease in hematocrit to 29.0 ± 0.5% was observed. The number of white blood cells was within the reference values and was 9.7 ± 0.5 *10^9/l. In the analysis of leukograms, neutrophilia was identified with the appearance of young cell forms, which suggests a possible course of the inflammatory process. Thus, it can be concluded that carrying out timely medical examination of fresh animals in violation of mineral metabolism will allow to assess the prospects for further use of cows and, taking the necessary measures, to extend their productive longevity.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
DESIGN AND SYNTHESIS OF A siRNA AGAINST ENDOTHELIN‐1 FOR THE TREATMENT OF PULMONARY ARTERIAL HYPERTENSION IN A MODEL INDUCED BY MONOCROTALINE IN RATS
Authors: Villafaña Santiago , Blancas Citlali Margarita
Published: June 2021

Pulmonary arterial hypertension (PAH) is an orphan disease characterized by an increase in pulmonary arterial pressure greater than 25 mmHg, in which patients have a poor prognosis. Among the mechanisms involved in this pathology is endothelin‐1 which is a polypeptide of 21 amino acids that participate in the regulation of vascular tone, the increase of endothelin‐1 causes vascular resistance in the pulmonary arteries, which elevates pulmonary arterial pressure for this reason, the aim of this work is to design and synthesize a siRNA against endothelin‐1 to reduce the vascular resistance and cardiac hypertrophy associated with PAH. Male Wistar rats were administered with monocrotaline (60 mg/kg i.p.) to induce PAH. After four weeks, the ratio of RV/LV + S as an index of right ventricular hypertrophy, mean arterial pressure of the right ventricle, blood pressure and weight were determined. For the siRNA's design, we used Wizard v3.1, RNAfold and EMBOSS software programs, the selected sequences were synthesized by a MERMADE 8. The results showed a significantly increase in the ratio of RV/LV + S, mean arterial pressure of the right ventricle and a decrease in the weight compared with the controls while the bioinformatic analysis of the mRNA suggests that 519–541 and 951–968 regions are susceptible to degrade. For this reason, we suggest that the siRNA against endothelin‐1 could be a therapeutic option for the treatment of PAH.

Support or Funding Information

Grants: Instituto Politécnico Nacional SIP‐20194983 and Comisión de Operación y Fomento de Actividades Académicas del IPN (COFAA)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Use of Recordings of Real Patients to Improve Clinical Integration of a First Year Medical Student Pulmonary Function Testing Laboratory
Authors: Sullivan Rebecca Petre , Zantah Massa , Patel Maulin , Fanrak Thomas , Garfield Jamie
Published: June 2021

The Lewis Katz School of Medicine at Temple University annually welcomes 200 students to the MD program. We aim to create an interactive and collaborative experience to promote deep levels of learning for large groups of students. The curriculum is being transitioned from a two‐pass to a single‐pass model (AY 2020–21) with early integration of the basic and clinical sciences.

The pulmonary function testing (PFT) laboratory in a first year organ systems course has been redeveloped. Previously, the lab utilized Collins spirometers to allow students to gain experience recording and determining lung volumes and capacities and discussing concepts related to lung function testing and respiratory system mechanics. Advantages of the lab included visualization of the mechanics of the test, and hands‐on data collection and analysis. Disadvantages included frequent technical difficulties with aged spirometers and the lack of direct observation of abnormal lung mechanics, and therefore a disconnect to the pathophysiology learned in a subsequent course.

Our goal was to address these issues while encouraging collaboration of basic science and clinical faculty, therefore better connecting physiology concepts with pathophysiologic mechanisms of disease. We redeveloped the lab to use the existing structure of calculations and discussion questions while adding the data and experiences of real patients from the Temple Lung Center who underwent modern pulmonary function testing. A patient with obstructive lung disease, a patient with restrictive lung disease, and a person with normal lung function, were recruited to participate in a video recording.

In the session, the students watch a video which includes an explanation of the equipment, review of the procedure, and review of lung volumes/capacities previously learned in lecture format. The footage includes patients performing the lung function testing alongside real time flow‐volume loops and volume‐time graphs. The patients with abnormal lung function are introduced in vignette form with radiographic images. Students discuss each case with faculty and predict what they expect to observe with regards to lung volumes/capacities before watching the patients perform the tests. Students are then asked to use and apply real data collected from the patient in the video with normal lung function and complete discussion questions in their small groups (6/group).

The reimagined PFT laboratory allows first year students the opportunity to apply basic science knowledge to real and relevant patient cases. This session allows students to achieve the learning objectives as per the APS Medical Curriculum Objectives Project (www.the‐aps.org/MedPhysObj).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Early prognostic capacity of serum lactate for severe postpartum hemorrhage (SPPH)
Authors: Kway Venance Basil , Reyther Roberto Arturo Castillo , Salgado Luis Andrés Domínguez , Tanguma Ricardo Espinosa , Moreno Úrsula Fabiola Medina , Moscoso Antonio Augusto Gordillo
Published: June 2021

Introduction

25% of maternal deaths worldwide are due to postpartum hemorrhage (PPH)1. Tissue hypoperfusion causes anaerobic respiration and lactic acid production. Lactate measurement has shown prognostic utility in different clinical conditions as severe sepsis and polytrauma.

Objective

To Determine whether the concentration of serum lactate measured at the diagnosis of postpartum hemorrhage indicates the development of severe hemorrhage.

Material and Methodology

A prognostic cohort study was carried out including 30 patients in vaginal labor and cesarean section who presented bleeding ≥ 500 ml measured by gravimetric method. A venous blood sample was taken for analysis of serum lactate. Those with a total loss of ≥ 1500 ml at the end of labor were classified with severe bleeding. The relationship between serum lactate elevation and development of severe hemorrhage (SPPH) was analyzed by chi‐square and odds ratio was calculated. The best cut point was determined by Curve‐ROC, and its prognostic capacity was compared with the cut‐off point of 2.2 mmol/l as used in ATLS.

Results

Thirty patients were analyzed. General average volume of bleeding: 1260 + 691 ml; in severe group was 1700 + 690 ml and 1100 + 224 ml in non‐severe, p = 0.0004. General average serum lactate: 2.68 + 1.8 mmol/l; in severe group 3.24 + 3 mmol/l vs non‐severe 2.28 + 0.54 mmol/l, p = 0.0120. The cut point that best differentiates the groups was 2.6 mmol/l, OR of 12.50 (1.34–116.8), p = 0.017; sensitivity: 0.71 (0.29–0.96); specificity 0.83 (0.52–0.98) and AUC = 0.74 for severe hemorrhage.

ATLS cut point of 2.2 mmol/l had RR of 3.65 (0.57,23.37), OR 6.55 (0.68, 63.33, 0.077 sensibility of 0.92 (0.64, 1.00), specificity of 0.35 (0.14, 0.62) positive predictive value of. 1.43 (0.97, 2.10)

Support or Funding Information

Consejo Nacional de Ciencia y Tecnología (CONACYT)

COMPARATIVE TABLE

Cut points
RR
OR
P VALOR
Sensibilidad
Especificidad
PPV

2.2
3.65 (0.57, 23.37)
6.55 (0.68, 63.33)
0.077
0.92 (0.64, 1.00)
0.35 (0.14, 0.62)
1.43 (0.97, 2.10)

2.6
5.50 (1.46, 20.71)
17.88 (2.73, 116.88)
<0.001***
0.85 (0.55, 0.98)
0.76 (0.50, 0.93)
3.60 (1.48, 8.74)

RR: RELATIVE RISK

OR: ODDS RATIO

PPV: POSITIVE PROGNOSTIC VALUE

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Estrogen Promotes Resolution of Pneumonia via CD4+Foxp3+ Tregs
Authors: Xiong Ye , Palmer Tsvi , Wang Lan , Zhong Qiong , Benner Alison , Damico Rachel , D'Alessio Franco Rafael
Published: June 2021

Pneumonia (PNA) is one of the leading causes of death worldwide. Current treatments for PNA are focused on the pathogens (e.g. vaccinations and antibiotics), but don't target excessive lung inflammation elicited by the host immune response; underscoring the need to identify additional therapeutic targets in PNA. Clinical and experimental evidence exists for sex differences in PNA outcomes and survival, with males having higher mortality. In contrast to studying early events (i.e., risk of infection and transmission), our work focuses on active immunoregulatory pathways that promote resolution of inflammation and lung repair with the ultimate goal to improve PNA‐ALI outcomes. Recognizing the critical role of Regulatory T cells (Tregs) in resolution of lung injury, we predict that improved PNA outcomes in females are the result of sex differences in Treg biology. We cultured Tregs and stimulated them with E2, our data showed exogenous estradiol (E2) promoted Treg suppressive phenotype and proliferation in vitro. We challenged male and female wild‐type mice with intratracheal Streptococcus pneumoniae to make murine model of PNA during early and resolution phases. Despite similar early lung injury, resolution was delayed in male mice compared to female while displaying increased lung CD4+Foxp3+ Tregs. To determine therapeutic effects of E2 on resolution, we rescued male mice 2 days after PNA with systemic administration of E2. Comparing with vehicle administrated mice, male mice after systemic administration of E2 had increased Tregs. The therapeutic effects of E2 were Treg‐dependent as Treg‐depleted (diphtheria toxin‐treated Foxp3DTR) mice did not resolve their injury in response to estrogen. Our results showed that estrogen augments Treg number and function to accelerate repair experimental PNA‐induced lung injury. Estrogen represents novel treatment for PNA by enhancing the host immune system.

Support or Funding Information

R01 Franco D'Alessio

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The effect of whole body cooling on cognitive processing
Authors: Sawai Norie , Kobayashi Fumino , Kubo Hiroko , Nakata Hiroki , Shibasaki Manabu
Published: June 2021

We previously demonstrated that heat stress reduced the peak amplitude of the P300 component of event‐related potentials (ERPs), but shortened or did not change the peak latencies of the P300 component and reaction time. Cold exposure is thought to decrease exercise or working performance, but the effect of body cooling on cognitive processing remains unknown. We hypothesized that whole‐body cooling decreases the peak amplitudes of the P300 component and delays its peak latencies. Fourteen healthy subjects, wearing a water‐perfused suit, performed somatosensory Go/No‐go paradigms twice (sessions 1 and 2) under two conditions (Cold and Control) on different days. Whole‐body cooling was achieved by circulating 20 °C water through the suit. Only session 2 under the Cold condition was performed during whole‐body cooling, while the other sessions were performed with circulating 34 °C water. Passive cold stress for 40 min decreased the mean skin temperature from 35.0 ± 0.5 °C (Session 1) to 30.4 ± 0.9 °C (Session 2), while the internal temperature (external canal temperature) was maintained. Passive cold stress delayed the peak latency of the Go‐P300 component on ERPs, while the reaction time did not change. The peak amplitudes of N140 and P300 on ERPs did not change. These results suggest that mild whole‐body cooling slows the nerve conduction velocity on higher cognitive functioning but does not change the amount of neural activity.

Support or Funding Information

JSPS 18H03166

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differential impact of severe familial hypercholesterolemia on regional skeletal muscle and organ blood flows during exercise: effects of PDE5 inhibition
Authors: Aragonez Christian G , de Beer Vincent J , Tharp Darla L , Bowles Douglas K , Laughlin M. Harold , Merkus Daphne , Duncker Dirk J , Bender Shawn B
Published: June 2021

Swine with familial hypercholesterolemia (FH) exhibit attenuated exercise‐induced systemic vasodilation that is restored by phosphodiesterase 5 (PDE5) inhibition. Whether the impacts of FH and PDE5 inhibition to impair and restore, respectively, exercise‐induced vasodilation results from tissue‐specific or generalized effects remains unclear. Thus, we hypothesized that FH induces generalized impairment of skeletal muscle vasodilation that would be alleviated by PDE5 inhibition. Systemic vascular responses to exercise were assessed in chronically instrumented normal and FH swine before and after PDE5 inhibition with EMD360527. Skeletal muscle and organ blood flows and conductances were determined via the microsphere technique. As previously reported, versus normal swine, FH swine have pronounced elevation of total cholesterol and impaired exercise‐induced vasodilation that is restored by PDE5 inhibition. Blood flows to several, not all, skeletal muscle vascular beds were severely impaired by FH associated with reduced blood flow to many visceral organs. PDE5 inhibition differentially impacted skeletal muscle and organ blood flows in normal and FH swine. These data indicate that FH induces regional, not generalized, vasomotor dysfunction and that FH and normal swine exhibit unique tissue blood flow responses to PDE5 inhibition thereby adding to accumulating evidence of vascular bed‐specific dysfunction in co‐morbid conditions.

Support or Funding Information

Supported by NIH (HL52490 to MHL, HL136386 to SBB), The Netherlands Cardio Vascular Research Initiative, an initiative with support of the Dutch Heart Foundation (CVON2014‐11, RECONNECT to DM and DJD), and the Department of Veterans Affairs BLR&D (CDA‐2 IK2 BX002030 to SBB).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Bolus Dose of PTH 1‐84 Elicited Sustained Elevations in Intramedullary Pressure in Rat Femora
Authors: NOH Sunggi
Published: June 2021

BACKGROUND

Previous studies demonstrated that chronic increases and decreases in bone intramedullary pressure (IMP) augments and reduces bone mass, respectively. Parathyroid hormone (PTH) plays a central role in the regulation of bone remodeling and augments bone mass. We previously reported augmented vasodilation of the femoral principal nutrient artery (PNA) to cumulative doses of PTH 1‐84, PTH 1‐34 and PTHrP 1‐34. In addition, bone perfusion is augmented following a bolus dose of PTH. Since PTH increases vasodilation and blood flow, we sought to assess its effects on bone IMP. The purpose of the study was to sequentially examine IMP in rat femora following PTH administration.

METHODS

Male Wistar rats (3–5 months) were randomly divided into the following groups: Control (CON, n = 7), PTH120 (n = 7), PTH150 (n = 7), and PTH330 (n = 7). Rats were anesthetized (2% isoflurane to O2 balance) and right femoral shafts were catheterized. At this time, the PTH groups received PTH 1‐84 (100 μg/kg, s.c.) and CON received PBS (100 μl, s.c.). The rats were sacrificed 120, 150 and 330 minutes following surgery. Just prior to sacrifice, the catheter was connected to a pressure transducer and in vivo IMP (mmHg) was recorded (PowerLab, AD Instruments) in the conscious rodents. The last 10 minutes of data were averaged and reported. Alpha level was set a priori at p<0.05 and One‐way ANOVA and Student Newman‐Keuls post hoc were performed to determine group differences.

RESULTS

Body mass did not differ among groups: CON, 478 ± 8 g; PTH120 474 ± 9 g; PTH150, 458 ± 6 g; PTH330, 488 ± 4 g. IMP was augmented (p<0.05) at PTH120 (33 ± 3 mmHg), PTH150 (35 ± 6 mmHg), PTH330 (41 ± 4 mmHg) vs. CON (20 ± 3 mmHg). IMP did not differ among the PTH groups.

CONCLUSION

PTH increased bone IMP and sustained this response for up to 5 1/2 hours (i.e., 330 min) following administration. This increase in pressure corresponds with reports of enhanced vasodilation and bone perfusion following bolus administration of PTH. Given the bone altering effects of chronic IMP modulation, enhanced IMP with PTH administration may represent another mechanism by which it induces bone anabolism.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Chemerin decreases vascular reactivity to contractile stimuli in resistance vessels from Sprague Dawley rats fed a high fat diet for 1 month
Authors: Klee Nicole , McKinzie Jaine , Arishe Olufunke , Priviero Fernanda , Webb Clinton
Published: June 2021

Obesity and cardiovascular disease (CVD) are comorbid with a common underlying pathophysiology. The adipokine chemerin positively correlates with both obesity and CVD and has been proposed as the link between fat and arterial contraction. Chemerin has been shown to augment vascular reactivity to contractile stimuli; however, the effects of a high fat diet (HFD) on chemerin vascular reactivity are currently unknown. We hypothesize that animals fed a HFD will have augmented vascular responses to chemerin compared to regular diet fed controls. We fed 13–15 week old Sprague Dawley rats with a HFD (45% kcal) or regular diet for 1 month [body weight (g), regular diet: 410 ± 4.573 vs. HFD: 443 ± 12.77; p<0.05). Vascular function was performed on isolated mesenteric resistance arteries (MRA), pudendal arteries, and aorta using wire and pin myographs. Vascular reactivity in response to a chemerin concentration response curve (10−9 M–10−5 M) was not significantly different between regular and HFD fed animals in any vascular bed we measured. Aortic vascular reactivity to phenylephrine (PE) in both regular and HFD was significantly augmented after 1 hour chemerin incubation (0.5 ng/mL and 5 ng/mL). However, after 1 month of HFD reactivity to PE was significantly decreased in resistance arteries after 1 hour incubation with chemerin (5 ng/mL) compared to vehicle [Force (mN) (PE 10−9 M–10−5 M), vehicle vs. chemerin; interaction pudendal p<0.05, MRA p<0.05]. This response was absent in the regular diet fed group [Force (mN) (PE 10−9 M–10−5 M), vehicle vs. chemerin; interaction pudendal p = 0.9497, MRA p = 0.9834].

Our data indicate that in regular diet fed male Sprague Dawley rats, chemerin augments vascular reactivity in conductance but not resistance vessels. After 1 month of a HFD, the effects of chemerin in resistance vessels decreased but did not change the augmented vascular reactivity seen with regular diet in conductance vessels. Therefore, our hypothesis that chemerin would augment vascular reactivity in animals fed a HFD was not supported by our results. It is important to note that longer term studies are needed.

Support or Funding Information

NIH PO1 HL‐134604

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Mouse Genetic Background influences Endothelial Function along the Mouse Vascular Tree
Authors: Holly Dylan Stone , Kim Hyoseon , Shin Song Yi , Dezell Hannah , Woodman Christopher R , Massett Michael P
Published: June 2021

The endothelium is critical for maintaining vascular integrity and protecting against cardiovascular disease. Endothelial dysfunction is associated with a host of disease states including cardiovascular disease, diabetes, chronic kidney disease, and Alzheimer's disease. However, it is unclear whether endothelial function or dysfunction varies throughout the vasculature. In addition, accumulating evidence suggests that endothelial function, and more generally vasomotor function, is genetically regulated. The purpose of this study was to determine the influence of genetic background on vasomotor function in blood vessels of differing size. Vasomotor function was measured in aorta (thoracic (TA), and abdominal (AA)) and conduit arteries (carotid artery (CA) and femoral artery (FA)) from two inbred strains (NZW/LacJ (NZW) and C57BL/6J (B6)) of mice. Arteries were dissected, cut into 2 mm segments, and mounted in a wire myograph system. Increasing concentrations of phenylephrine (PE, 1×10−9 – 1×10−5M) were used to measure contractile responses, while increasing concentrations of the endothelium‐dependent vasodilator acetylcholine (ACh, 1×10−9 – 1×10−5M) and endothelium‐independent vasodilator sodium nitroprusside (SNP, 1×10−9 – 1×10−5M) were used to assess relaxation responses. Contractile responses to PE were not significantly different between strains across all vessels. Endothelium‐dependent relaxation to ACh was significantly impaired in TA (P = .0001), AA (P = .0019), and FA (P = .0002) from NZW as compared to B6, while CA relaxation responses were not statistically different between groups. Sensitivity to ACh as determined by IC50 was impaired in NZW TA (P = .0006), AA (P = .0005), CA (P = .001), and FA (P = .0003). Relaxation responses to the endothelium‐independent vasodilator SNP were not statistically different in the AA, CA and FA, while TA relaxation to SNP was statistically lower in B6 TA compared to NZW (P = .02). Sensitivity to SNP was not statistically different between groups for TA, CA, and FA but NZW AA showed a greater sensitivity to SNP than did B6 (P = .0304). These findings indicate a strong influence of genetic background on vasomotor function and could be important in understanding the implications that genetics have on vascular response to vasoactive agents.

Support or Funding Information

This work was supported by a Texas A&M Triads for Transformation grant (Michael Massett, Christopher Woodman), a Texas A&M Merit Fellowship (Dylan Holly), and a Texas A&M College of Education Strategic Research Award (Song Yi Shin).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
MicroRNA Profiling in Coronary Endothelial Cells in Type 2 Diabetic Mice
Authors: Zhang Qian , Tsuji‐Hosokawa Atsumi , Watanabe Makiko , Makino Ayako
Published: June 2021

Coronary endothelial dysfunction is a major cause of cardiac ischemia and it is commonly seen in patients with diabetes. MicroRNAs (miRNAs) are small noncoding RNAs that modulate the expression of multiple mRNAs. There is increasing evidence showing that the change in miRNA expression is implicated in the development and/or progression of cardiovascular diseases. However, the pathological role of miRNAs in coronary endothelial dysfunction in type 2 diabetes (T2D) is still unknown. MicroRNA array was used to profile the expression level of miRNAs in coronary endothelial cells (ECs) isolated from control and T2D mice. Real time PCR analysis was performed to confirm the miRNA array results. Among 491 murine miRNAs tested, 64 up‐regulated and 20 down‐regulated miRNAs were identified and seven miRNAs (miR106b, 210, 221, 222, 342‐5p, 378, 705, fold change>1.25) were selected to confirm the data using real‐time PCR. Real time PCR data revealed that miR378 expression was significantly higher and miR210 and miR342‐5p expression levels were significantly lower in diabetic ECs than in control ECs. To identify the target genes of miR378 and 342‐5p, we transfected miR378 mimic or miRNA342‐5p inhibitor in mouse coronary ECs. We chose 91 genes that are highly expressed in ECs and play crucial roles in endothelial functions, including a) endothelium‐derived relaxing factors and their regulators; b) modifiers of cytosolic, mitochondrial, and endoplasmic Ca2+ concentration; and c) regulators of EC proliferation/migration/apoptosis. The real‐time PCR data showed that miR378 mimic overexpression altered mRNA levels of 11 genes and miR342‐5p inhibitor in 5 genes. Among those genes, Vegfb, Atp3a3, Stim2 were decreased in coronary ECs transfected with miRNA mimic or inhibitor and in coronary ECs in T2D mice compared to their controls. These data suggest that miR378 and/or miR342‐5p are a potential target to restore coronary endothelial dysfunction in diabetes.

Support or Funding Information

This work was supported by a grant from the National Heart, Lung, and Blood Institute of the National Institutes of Health (HL142214 to A. Makino).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Lower baroreflex sensitivity during muscle metaboreflex activation in children and adolescents with the Fontan circulation and single ventricle physiology
Authors: Boyes Natasha G , Iqbal Ramlah M , Fusnik Stephanie , Lahti Dana S , Erlandson Marta C , Wright Kristi D , Kakadekar Ashok , Bradley Timothy J , Pharis Scott , Pockett Charissa , Olver T Dylan , Tomczak Corey R
Published: June 2021

Individuals with the Fontan circulation and single ventricle physiology have elevated sympathetic nerve activity at rest and during muscle metaboreflex activation that is consistent with autonomic dysfunction. Adults with the Fontan circulation also show reduced baroreflex sensitivity (BRS); however, there is limited understanding of BRS in children or adolescents with the Fontan circulation. In healthy adults, BRS decreases with increased sympathetic nerve activity during isometric handgrip exercise, but not during isolated muscle metaboreflex activation. The effect of handgrip exercise and muscle metaboreflex activation on BRS in children and adolescents with the Fontan circulation is currently unknown.

PURPOSE

We tested the hypothesis that BRS is lower at rest and less responsive during handgrip exercise and post‐exercise circulatory occlusion (PECO) in children/adolescents with the Fontan circulation compared to healthy controls.

METHODS

Eight children/adolescents with the Fontan circulation (12 ± 2 yrs; 3 males) and 8 healthy controls (13 ± 4 yrs; 5 males) were studied. Continuous heart rate (ECG) and non‐invasive blood pressure (Finometer® MIDI) were recorded during 3‐min of resting baseline, 2‐min of isometric handgrip exercise at 30% of maximal voluntary contraction, and 3‐min of PECO. Continuous heart rate, systolic blood pressure, and diastolic blood pressure signals were analyzed using the BRS Analysis software (Nevrokard, Slovenia, 2018) to yield average BRS values during rest, handgrip, and PECO for each group. Differences between groups and across conditions were assessed using two‐way repeated measures (2 × 3, group × condition) ANOVA. P<0.05 was considered significant.

RESULTS

There was a significant group × condition interaction for BRS (P<0.001). BRS was lower in children/adolescents with the Fontan circulation vs. healthy controls at rest (13 ± 8 vs. 38 ± 10 ms/mmHg; P<0.001) and during PECO (18 ± 17 vs. 37 ± 18 ms/mmHg; P=0.044), but not during handgrip (13 ± 10 vs. 14 ± 6 ms/mmHg; P=0.843). BRS in children/adolescents with the Fontan circulation did not change across conditions (P=0.188).

CONCLUSION

Our findings suggest that BRS is lower at rest and has a blunted response during sympatho‐excitation in children/adolescents with the Fontan circulation compared to healthy controls. Collectively, our findings indicate potential autonomic dysfunction in children/adolescents with the Fontan circulation.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Determination of the Role of LOX‐1 during Escherichia coli Pneumonia
Authors: Molina William , Korkmaz Filiz , Symer Elyse , Jones Matthew , Mizgerd Joseph , Traber Katrina , Quinton Lee
Published: June 2021

Pneumonia is primarily caused by infection from bacteria, virus, and/or fungi and results in an acute lower respiratory tract inflammation. It is responsible for severe morbidity and mortality worldwide. Pneumonia is the foremost cause of children's hospitalizations in the United States and accounts for more than 935,000 deaths globally. In mice with severe bacterial pneumonia it is known that the pathophysiology of pneumonia is due to the overt activation and recruitment of host innate immune cells. Data generated in our lab has shown that there is an induction of the OLR1 gene that expresses lectin‐like oxidized low‐density lipoprotein receptor 1 (LOX‐1) in the lungs of mice during pneumonia. LOX‐1 is a receptor that is expressed in many cells, including host innate immune cells and it is commonly associated with atherosclerosis and inflammation. Herein, we aim to understand the role LOX‐1 plays during Gram‐negative bacterial pneumonia. Our recent results indicate that local neutralization of LOX‐1 within the lungs during pneumonia caused a significant increase in total tissue injury and inflammatory cytokines, indicating that LOX‐1 may have a protective effect.

To determine the effect of LOX‐1 inhibition systemically, an intraperitoneal injection with 10 μg of anti‐LOX‐1 antibody was administered to neutralize LOX‐1 in C57BL/6J mice that were subsequently infected with Escherichia coli. Mice were euthanized 24 hours after infection and bronchoalveolar lavage fluid (BALF) samples were collected. In an ex vivo system, mouse lung epithelial (MLE‐12) cells were similarly treated with anti‐LOX‐1 antibody to observe the effect of neutralizing LOX‐1 on epithelial cell death. BALF from infected and uninfected mice was also added in the ex vivo model to more accurately recreate the conditions of the alveolar space. Total protein from MLE‐12 cells was then collected 24 hours after stimulation for analysis and characterization.

Overall, the systemic neutralization of LOX‐1 in C57BL/6J mice during bacterial pneumonia did not significantly affect leukocyte recruitment or lung injury. In addition, ex vivo LOX‐1 neutralization in MLE‐12 cells did not affect apoptosis as measured by cleaved caspase‐3 on a western blot. In contrast with local inhibition of LOX‐1 in the lungs, systemic inhibition of LOX‐1 did not significantly limit tissue injury in mice with pneumonia. Blockade of LOX‐1 in ex vivo model did not remarkably affect epithelial cell death or apoptosis. Further studies involving measurement of resistance and resilience of lung tissue are underway and will more completely characterize the role of LOX‐1 during pneumonia.

Support or Funding Information

NIH grants R01‐HL111449, R01‐GM120060, and T32‐HL007035.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sex Differences in Systemic and Renal Tissue Urea Concentrations
Authors: Boesen Erika Ingrid , Wang‐France Jun , Rahman Shamma Shakila
Published: June 2021

Inner medullary urea accumulation is believed to play a key role in determining urinary concentrating ability. We had previously studied the effects of different levels of dietary protein concentration on renal excretory function in mice as a test of renal urea handling. Unexpectedly, we observed an increase of urine flow in male but not female mice when switched to a low protein diet (6% protein). To further investigate this phenomenon, we fed diets containing either a low (6%) or normal (20%) protein content to male and female mice for 7 days (n=7–10/group), followed by collection of plasma and renal tissue for measurement of osmolarity and urea concentration. Plasma osmolarity was significantly lower in female compared with male mice irrespective of diet (PSex< 0.05), and both sexes had significantly lower plasma osmolarities on 6% compared with 20% protein diets (by ~5 mOsmol/kg. H2O; PDiet<0.001). Dietary protein levels had a significant effect on plasma urea concentration irrespective of sex, with urea concentrations being lower on 6% compared with 20% protein diet (PDiet< 0.0001). Plasma urea levels were also significantly lower in female compared to male mice independent of diet (PSex<0.001). The effect of dietary protein intake varied by sex (PSex*Diet<0.01), with plasma urea concentration significantly higher in male compared to female on 20% protein diet (P<0.05 by post‐hoc test; male 9.7 ± 0.3 mM, female 6.8 ± 0.5 mM) but not on 6% protein diet (P>0.05; male 4.9 ± 0.3 mM, female 4.5 ± 0.1 mM). Inner medullary tissue osmolarity was significantly lower on 6% compared to 20% protein diet, independent of sex (PDiet<0.01), but there was no significant sex difference (PSex=0.8) or difference in the way the sexes responded to diet (PSex*Diet=0.8). Dietary protein levels also had a significant effect on inner medullary urea concentration, being lower on 6% compared to 20% protein diet in both male and female mice (PDiet<0.01). Further, there was a sex difference in inner medullary tissue urea concentration, with male mice showing lower urea concentrations compared to female mice, independent of level of dietary protein content (PSex=0.05). There were no significant effects of sex or diet on outer medullary or renal cortical tissue osmolarity, or on outer medullary tissue urea concentration. Renal cortical tissue urea concentration was significantly higher in female compared to male mice, independently of diet (PSex<0.01), but there no significant effect of the level of dietary protein content (PDiet=0.07). Together, our data indicate that there are sex differences in circulating and renal tissue urea concentrations, which we speculate may contribute to sex differences in the effect of dietary protein intake on urinary concentrating ability.

Support or Funding Information

American Heart Association 17GRNT33661008

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Physical and Metabolic Characteristics of p62 Knockout Mouse: A New Animal Model of Obesity/Insulin Resistance
Authors: Kim Do‐Houn , Salazar Gloria , Hwang Hyun‐Seok , Klemp Alex , Yeh Ming‐Chia , Worts Phillip R. , Panton Lynn B. , Arjmandi Bahram H. , Grant Samuel C. , Kim Jeong‐Su
Published: June 2021

Objectives

The p62 deficient mouse exhibits a similar but unique phenotype of obesity compared to other diet‐induced or genetically modified obesity models. Our study aimed to further characterize physical and metabolic properties of p62 knockout mice.

Methods

Male C57BL/6 p62 knockout (p62−/−, 14 weeks old, n=9) and sex‐/age‐matched C57BL/6 wildtype control mice (CON, n=9) were studied. All mice were raised separately with access to standard rodent chow and water ad libitum. Food intake and body weight changes were monitored biweekly for 20 weeks. At 24 and 34 weeks of age, all mice went through in vivo assessments of body composition (dual X‐ray absorptiometry), neuromuscular function (grip strength and inclined plane test), and oral glucose tolerance tests (OGTT). Additionally, at 34 weeks of age, hindlimb muscles (i.e., gastrocnemius, soleus, and plantaris) and the spleen were extracted and weighed. The isolated muscles were analyzed for myofiber cross‐sectional area (CSA) and expression levels of regulatory proteins for muscle growth (i.e., mTOR, AKT, atrogin1, and MuRF1), insulin signaling [i.e., insulin receptor β (IR‐β) and GLUT4], and proinflammation (i.e., TNF‐α).

Results

p62−/− demonstrated greater daily food intake and weight gain leading to obesity at 22 weeks of age compared to CON (p<0.05). At 24 weeks of age, p62−/− displayed greater total body mass and fat mass (p<0.05), and these changes were even greater at 34 weeks of age. p62−/− showed higher fasting blood glucose levels and diminished blood glucose clearance leading to greater postprandial blood glucose (p<0.05). No significant differences in grip strength and sensorimotor function were observed in p62−/− compared to CON. In vitro analysis revealed no group difference in muscle wet weights; however, spleen weights were greater in p62−/− than CON. p62−/− exhibited greater myofiber CSA than CON (p<0.05). There was a suppressed expression ratio of phosphorylated AKT to total AKT in the gastrocnemius of p62−/− (p<0.05).

Conclusion

Young adult p62−/− mice developed obesity accompanied by insulin resistance and systemic inflammation, which was attributable, at least in part, to increased food intake. However, it is noteworthy that p62−/− mice exhibited greater myofiber CSA while maintaining skeletal muscle mass and function during progression of obesity.

Support or Funding Information

Jiwon Co. Ltd

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A thermogenic‐like brown adipose tissue phenotype is dispensable for enhanced glucose tolerance in female mice
Authors: Winn Nathan , Acin‐Perez Rebeca , Woodford Makenzie , Hansen Sarah , Haney Megan , Ayedun Lolade , Rector R. Scott , Vieira‐Potter Victoria , Shirihai Orian , Sacks Harold , Kanaley Jill , Padilla Jaume
Published: June 2021

The prevailing dogma is that thermogenic brown adipose tissue (BAT) contributes to improvements in glucose homeostasis in obesogenic animal models; though, much of the evidence supporting this premise is from thermostressed rodents. Whether modulation of the BAT morphology/function drives changes in glucoregulation at thermoneutrality requires further investigation. We utilized loss and gain‐of‐function approaches including genetic manipulation of the lipolytic enzyme Pnpla2, change in environmental temperature, and lifestyle interventions to comprehensively test the premise that a thermogenic‐like BAT phenotype is coupled with enhanced glucose tolerance in female mice. In contrast to this hypothesis, we found that: (i) compared to mice kept at thermoneutrality, enhanced activation of BAT and its thermogenic phenotype via chronic mild cold stress does not improve glucose tolerance in obese mice; (ii) silencing of the Pnpla2 in interscapular BAT causes a brown‐to‐white phenotypic shift but does not disturb glucose tolerance in lean mice; and (iii) exercise and low‐fat diet improve glucose tolerance in obese mice but these effects do not track with a thermogenic BAT phenotype. Collectively, these findings indicate that a thermogenic‐like BAT phenotype is not linked to heightened glucose tolerance in female mice.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
hNAG‐1 improves insulin sensitivity and ovarian function in young and aged female mice upon high fat diet treatment
Authors: wang Yujie , Wu Kaikai , Guo Dandan , Guo Chenjie , Pan Haitao , Sang Tingting , Wang Ying , Wang Xingya
Published: June 2021

Nonsteroidal anti‐inflammatory drug‐activated gene‐1 (NAG‐1) is a distant member of the TGF‐β superfamily. Female hNAG‐1 transgenic mice are resistant to high fat diet (HFD)‐induced obesity and have increased lifespan. Progressive gonadal dysfunction has been shown to be an important sign of aging in experimental animals. Whether hNAG‐1 transgenic mice have improved ovarian function has not been studied. We examined the ovarian function in both young (20–28week) and old (50–56 week) female mice upon feeding with high or low fat diets for 16 weeks. We found that ovarian function is improved in hNAG‐1 mice compared to wild type littermates upon both diets, with the most prominent effects observed in older mice. We observed decreased body weight, abdominal fat and lower serum glucose, leptin, insulin and insulin‐like growth factor‐1(IGF‐1) levels, and increased steroid production and increased insulin sensitivity in hNAG‐1 mice compared to wild type mice. These observations were correlated with deregulation of the expression of pro‐inflammatory cytokines and follicular development regulatory genes that suggesting improved ovarian function in hNAG‐1 mice. Histological analysis of the ovaries showed a decrease in the number of primary and secondary follicles in both genotypes upon HFD treatment. However, the ovaries from wild type mice showed an increase in the number of hemorrhagic follicles compared to hNAG‐1 mice, suggesting that hNAG‐1 may improve ovarian function through preserving primordial follicles and increasing oocyte quality. In addition, western blot analysis demonstrated that expression of mTOR/IGF‐1/TLR4 signaling pathways were decreased in young and old hNAG‐1 mice ovaries, and an increase of Foxo/SIRT1 signaling pathway was observed in old hNAG‐1 mice. Taken together, the present results suggested that hNAG‐1 play an important role in improving ovarian function and insulin sensitivity in female

Support or Funding Information

National Natural Science Foundation of China (Grant No. 81400822).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Impaired Acute Hypoxic Sensing in Olfactory Receptor 78 Knockout Mice
Authors: Peng Ying‐Jie , Nanduri Jayasri , Wang Ning , Girdina Anna , Fox Aaron , Prabhakar Nanduri R
Published: June 2021

Chang et al. (2015) reported that olfactory receptor 78 knockout (Olfr78 KO) mice exhibit impaired breathing and carotid body (CB) responses to hypoxia. However, a recent study questioned this finding (Lopez‐Barneo et al. 2018). To resolve the controversy, present study was performed on age and gender matched wild‐type (WT) and Olfr78 KO mice on Bl6 and Jax background strains (n=10 mice each genotype). Breathing was monitored in conscious mice by plethysmography and expressed as minute ventilation; VE/VO2, as well as by recording phrenic nerve activity (minute neural respiration; MNR, AU) in anesthetized mice. Hypoxia (12%O2)‐evoked increase in VE/VO2 and MNR were significantly attenuated in Olfr78 KO mice on Bl6 and Jax backgrounds, respectively as compared to WT mice (p<0.01). Hypoxia (pO2 ~35mmHg)–induced CB sensory excitation was 61 and 43% less in Olfr78‐Bl6 and Jax, respectively as compared to WT mice (p<0.01), and this effect was associated with 78% reduction in [Ca2+]i response of CB glomus cells to hypoxia (p< 0.01). In contrast, breathing and CB sensory nerve responses to 10%CO2 were comparable between Olfr78 KO (both Bl6 and Jax) and WT mice. These findings demonstrate that Olfr78 KO mice have impaired breathing, CB sensory, and glomus cell responses to acute hypoxia.

Support or Funding Information

Supported by NIH‐HLBI P01‐HL‐90554

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of the Rho‐kinase (ROCK) pathway improves morphology and stress granule formation in ALS4 patient iPSC‐derived motor neurons
Authors: Amaya Joshua , Grunseich Christopher , Fischbeck Kenneth
Published: June 2021

Amyotrophic lateral sclerosis type 4 (ALS4) is an autosomal dominant motor neuron disorder caused by mutation in the senataxin gene (SETX), which is associated with upregulation of the TGFβ signaling pathway. ALS4 is characterized by early onset motor weakness, with distal muscle predominance and slow progression. The mechanism by which TGFβ pathway disruption may lead to degeneration of motor neurons is not clear. We used patient‐derived iPSC‐motor neurons to study the neuronal phenotype of ALS4. After 7 days of differentiation, ALS4 patient motor neuron‐like cells were found to have less morphological complexity, as defined by fewer branch points per soma and shorter axonal lengths, and this was reversed with Rho‐kinase inhibitor (RI) treatment. Morphological characterization was done using in vitro live cell imaging. In addition, stress granule formation, as quantified by immunofluorescence staining with G3BP antibody, was increased in ALS4 cells compared to controls. Analysis was done after 7 days of motor neuron differentiation. In summary, these results suggest that Rho‐kinase pathway inhibition can rescue abnormal morphological features in ALS4 patient‐derived motor neurons. In addition, we found that exogenous TGFβ treatment of motor neurons for 24 hours decreases the soma count in both patients and controls, recapitulating the toxicity of TGFβ stimulation. Understanding how the ROCK pathway modulates motor neuron biology and TGFβ upregulation may provide insight into the mechanism of the disease.

Support or Funding Information

National Institute of Neurological Disorders and Stroke (NINDS) intramural funding.

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