bluuebunny/crossref_metadata_2025 huggingface dataset:
Hypercapnic ventilatory response (HcVR) is increased in a rat model of Alzheimer's disease
Authors: Vicente Mariane C. , Bicego Kenia C. , Carrettiero Daniel C. , Almeida Maria Camila , Gargaglioni Luciane H.
Published: June 2021

Besides the typical cognitive decline, patients with Alzheimer's disease (AD) develop disorders in the respiratory system such as sleep apnea, shortness of breath and arrhythmias. These symptoms are aggravated with progression of the disease. However, the cause and nature of this disturbance are not well understood. Here, we, used an animal model for sporadic Alzheimer's disease (AD) induced by icv injection of streptozotocin (STZ) (2 mg/kg). We measured ventilation (VE) EEG and EMG during normocapnia, hypercapnia and hypoxia. In addition, we performed Western blot for phosphorylated Tau and β‐amyloid peptide (Aβ) in Locus Coeruleus (LC), Retrotrapezoid nucleus, Raphe Bulbar, Botzinger and Hippocampus and evaluated memory and learning acquisition by using the Barnes maze. STZ treatment promoted memory and learning deficits and an increase of 70% the of Aβ in the LC. STZ treated rats presented an increased HcVR during wakefulness (VE: STZ= 2456.7 ± 261.5 vs VE Vehicle= 1677.4 ±141.8 mL. Kg−1. min−1) and sleep (VE: STZ= (2106.9 ± 261.6 vs VE Vehicle= 1724.1 ± 37.4 mL. Kg−1. min−1). Our data suggest that sporadic AD animals present an increased sensitivity to CO2 possibly due an increased Aβ in the LC.

Support or Funding Information

São Paulo Research Foundation (FAPESP) and National Council for Scientific and Technological Development (CNPq)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Laminin‐β6 integrin Interaction is Crucial for Coronary Collateral Growth
Authors: Joseph Gregory , D'Addario Catherine , McEvoy Kyle , Jadhav Rashmi , Hutcheson Brenda , Rocic Petra
Published: June 2021

Extracellular matrix (ECM) composition and regulation is an essential component of coronary collateral growth (CCG). The ECM serves several functions which include mechanical and structural support, cell‐to‐cell communication, and regulation of cell mobility and phenotype. Laminin is a major component of the ECM. In the metabolic syndrome, the ECM composition is altered. In this study, we investigated the role of laminin and its binding to the β6 integrin in the regulation of CCG in response to repetitive ischemia (RI), and cellular processes involved in CCG. Coronary blood flow in Sprague‐Dawley (SD) or metabolic syndrome rats (JCR:LA‐cp, JCR), was measured in the collateral (CZ) and normal zone (NZ) of the heart using microspheres. Collateral‐dependent blood flow in JCR rats was impaired compared to SD rats (CZ/NZ flow ratio was 0.11±0.03 in JCR vs. 0.85±0.04 in SD), which correlated with decreased expression of laminin (~5 fold) and of the β6 integrin (~6 fold) in JCR animals. Administration of blocking‐antibodies against laminin or the β6 integrin in SD rats completely blocked CCG and decreased each other's expression, respectively. Furthermore, both laminin and β6 integrin expression were increased in endothelial cells (ECs) subjected to cycles of hypoxia‐hyperoxia‐normoxia to mimic RI vs. ECs cultured in normoxia. Blocking antibodies against laminin or the β6 integrin significantly impaired EC proliferation, migration and tube formation (~2–2.5 fold). TGF‐β signaling through Smad2/3 and leading to MMP 2/9 production, which is required for successful CCG, was also impaired (90%). Accordingly, TGF‐β receptor inhibition completely blocked CCG. Taken together, these results point to an important TGF‐β‐mediated pathway which promotes CCG and the activation of which is dependent on expression and the interaction between laminin and the β6 integrin.

Support or Funding Information

Support: NIH R01 HL093052

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Preconception Exposure of Particulate Matter Leads to Adult Cardiac Dysfunction through Altering Myocyte Function and Ca2+ Signaling Pathways
Authors: tanwar vineeta , Adelstein Jeremy , Sugar Benjamin , Schwieterman Neill , Youtz Dane , Falvo Michael , Wold Loren
Published: June 2021

Objective

Previous studies have demonstrated that particulate matter, or ambient particles less than 2.5 μm (PM2.5) in diameter, exposure during both in utero and postnatal developmental periods triggers electrical remodeling and cardiac dysfunction during adulthood. This shows that PM2.5 can reprogram hearts during the gestational period. Despite this evidence, cardiac effects from pre‐gestational particulate matter exposure remain inconclusive. This study was performed to further investigate the potential priming effects of preconception exposure of PM2.5 on global cardiac dysfunction at adulthood.

METHODS

Male and female FVB mice were exposed separately to either filtered air (FA) or PM2.5 at a concentration (within the annual average range of 15 μg/m3 according to the National Ambient Air Quality Standards (NAAQS)) for 3 months. Mice were then cross bred into two groups: (1) FAm X FAf (both parents were FA exposed) and, (2) PMm X PMf (both parents were PM exposed). Offspring born to these crosses (PC FA and PC PM2.5) were analyzed at 3 months of age for in vivo cardiac function via echocardiography, followed by in vitro cardiomyocyte functional and molecular analyses.

RESULTS

Echocardiography identified increased LVESd (2.58 ± 0.13 PC FA, 2.93 ± 0.13 PC PM2.5, P=0.1) and reduced PWTs (1.70 ± 0.06 PC FA, 1.44 ± 0.08 PC PM2.5, P=0.05) dimensions in PC PM2.5‐exposed mice. Morphological alterations were associated with lower systolic function indicated by reduced fractional shortening % (35.09% ± 1.34 PC FA, 29.05% ± 1.25 PC PM2.5, P=0.03) and ejection fraction % (64.86% ± 1.76 PC FA, 56.27% ± 1.95 PC PM2.5, P=0.03) in PM2.5‐exposed mice. Cardiomyocytes isolated from PC PM2.5 mice showed reduced peak shortening %PS (12.93% ± 0.42 PC FA, 11.10% ± 0.41 PC PM2.5, P=0.002), −dL/dT (−10.60 ± 0.82 PC FA, −8.50 ± 0.60 PC PM2.5, P=0.05), TPS90 (0.08 ± 0.003 PC FA, 0.07 ± 0.002 PC PM2.5, P=0.05) and slower calcium reuptake (tau, 0.42±0.06 s FA, 0.67±0.09 s PC PM2.5, P=0.05). qPCR analyses revealed decreased MURC, MyBPC3 and increased Mypt1 expression and western blot analyses demonstrated modified NCX, SERCA and PLN expression in PC PM2.5‐exposed mice compared to PC FA‐exposed mice.

CONCLUSION

Similar to our previous study involving in utero exposure, preconception exposure to PC PM2.5 at real‐world concentrations results in adult cardiac dysfunction. These results suggest that abnormalities in developmental potential are not limited to prenatal or postnatal period but can also be determined prior to conception.

Support or Funding Information

This work is supported by funding from National Institutes of Health (RO1ES019923) to LEW.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Angiotensin‐II‐Induced Increase in Blood Pressure in Female and Male Mice
Authors: Rouch Al , Platt Anna , Fan Liming
Published: June 2021

The kidney regulates blood pressure by controlling urinary salt and water excretion. Angiotensin II (AngII), a key player in controlling blood pressure, constricts blood vessels, stimulates vasopressin and aldosterone release, stimulates thirst, and increases sodium absorption from the proximal tubule. All effects cause increased blood pressure. Sex differences in the regulation of arterial blood pressure involve events surrounding the renin‐angiotensin‐aldosterone system. Evidence generally supports a protective mechanism for females with AngII‐induced hypertension; although, conflicting evidence exists. The purpose of this study was to investigate sex differences in CD‐1 mice under AngII administration for a period of 10 days. We hypothesized that female mice would be protected from AngII‐induced increase in mean arterial pressure (MAP) compared to male mice and that the renal handling of salt and water contributes to this protection. Four‐week old CD‐1 mice were obtained from Envigo, Inc. (Indianapolis, IN). Two studies, one with females and one with males, were conducted. Mice were placed in metabolic cages and consumed normal diet and water, ad libitum, for 15 days. After the first 5 days, an Alzet™ minipump containing either AngII (n=5) or vehicle (n=5) was implanted subcutaneously in each mouse to ensure continuous AngII infusion (1 mg/kg/min). Daily measurements included MAP via the tail‐cuff method (Kent Scientific, CODA System), urinary output, and urinary sodium excretion. MAP (mmHg) was not different between females and males in the 5‐day control period (85.1 ± 3.7 vs 81.7 ± 4.0, respectively). AngII‐treatment increased MAP in both females and males (110.3 ± 4.4 and 105.8 ± 3.0, p<0.001, compared to respective control period). Interestingly, AngII‐treated females showed higher urine output than AngII‐treated males (9.7 ± 0.8 vs 7.4 ± 0.4 ml/day, p<0.05) and also had higher sodium excretion (242 mEq/day vs 166 mEq/day, p<0.01). Absolute food intake in females and males was not different, however, females consumed more food per body weight. We conclude that female mice are not protected from AngII‐induced increase in MAP compared to male mice for a 10‐day AngII infusion period. However, based on the sodium excretion data, protection may be observed later from increased urine and sodium output in females. Results suggest that the renal handling of salt and water could be an important determinant of sex differences in blood pressure regulation.

Support or Funding Information

National Institute of General Medical Sciences of National Institutes of Health through Grant Number 8P20GM103447

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Muscle mitochondrial dysfunction at different stages of chronic kidney disease (CKD)
Authors: Gamboa Jorge , Keller Chad A. , Falck Aaron M. , Roshanravan Baback , Brown Nancy J. , Ikizler Talat Alp
Published: June 2021

Sarcopenia and frailty are commonly observed in patients with CKD. Mitochondria, as the principal source of energy, are important for proper muscle function, and may play a role in frailty and sarcopenia. We have previously reported reduced mitochondrial content in patients with CKD on maintenance hemodialysis (MHD) probably due to increased mitophagy. We have also found smaller mitochondria in skeletal muscle of patients on MHD, which may be the consequence of increased mitochondrial fission. We now evaluated the hypothesis that mitochondrial dysfunction is associated with the severity of chronic kidney disease (CKD). For this purpose we evaluated 22 patients on MHD, 20 patients with history of CKD stage 3–5 but not on MHD, and 21 controls with no history of CKD that were matched by gender, history of diabetes, BMI, and race. We measured in vivo mitochondrial function by 31‐phosphorus magnetic resonance spectroscopy (31P‐MRS) in the quadriceps muscle. We used the half time recovery of phosphocreatine (P‐Cr) after a brief exercise as the measure of mitochondrial function. A faster recovery correlates with better mitochondrial function. We also measured physical activity using the six‐minute walk test. Controls and patients on MHD were similar in terms of age (46.9±9.5 vs. 47.7±11.7) but younger compared to patients with CKD stage 3–5 (63.6±9.0). BMI was similar among the groups (30.1±5.9, 30.4±5.2, and 29.6±7.9; controls CKD stage 3–5, and patients on MHD, respectively). We found that the half time recovery of P‐Cr was faster in controls compared to patients on MHD, but similar to patients with CKD stage 3–5 (Figure 1A). We also found that physical activity measured by 6 minute walk test was higher in controls compared to patients with CKD stage 3–5 and patients on MHD (Figure 1B). There was significant negative correlation between physical activity and mitochondrial function (R2=0.37, p<0.001). Our data suggest that in vivo mitochondrial function is impaired in patients on MHD but not in patients with CKD stage 3–5 despite a progressive decrease in physical activity among the groups. Further studies are required to evaluate the efficacy of interventions, such as mitochondrial targeted‐therapies or exercise, on mitochondrial function in patients with CKD.

Support or Funding Information

NIH/NIDDK (K23DK100533)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Indispensably evil! The role of oxygen in nitric‐oxide dependent endothelial function
Authors: Pedrinolla Anna , Savoldelli Aldo , Scafidas Spyros , Giuriato Gaia , Fornasiero Alessandro , Pellegrini Barbara , Bortolan Lorenzo , Schena Federico , Venturelli Massimo
Published: June 2021

Background

Endothelium derived nitric oxide (NO) is crucial in preserving vascular tone. Several evidence shows that NO bioavailability and utilization is regulated by oxygen (O2) concentration. However, there are still not sufficient experimental data to show how O2 concentration affects NO‐dependent endothelial function in vasculature.

Aim

To investigate the role of O2 concentration on NO‐dependent endothelial function in vasculature. We hypothesized that different O2 concentration would have elicited different responses during single passive limb movement (sPLM) test, which is known to be an index of NO‐dependent systemic endothelial function.

Methods

Endothelium‐dependent dilation was measured in 7 active men (age, 34±12 years; weight, 70±8 kg, height, 173±4 cm, VO2max 55.9±10.5 ml/min/kg) by means of sPLM test performed in a normobaric‐hypoxic chamber in two conditions: Normoxic (N; FiO2 ~20.9%), and Hypoxic (H; FiO2 ~16.25%). For the two conditions femoral artery diameter, as well as basal blood flow (BF), Peak BF, ΔPeak BF, area under the curve (AUC) were calculated.

Results

In H, femoral artery diameter did not change (−2.2±3.7%, p=0.675), basal BF significantly increased (+24±9%, p=0.043), as well as peak BF (+32±10%, p=0.034), ΔPeak BF (+34±15%, p=0.040), and AUC (+44±16%, p=0.018).

Conclusion

As hypotized, different O2 concentrations provoked different responses during the sPLM test. These results suggest an O2‐dependent NO availability and utilization, which may play an essential role in endothelial function during hypoxia.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Role of Interleukin‐2 (IL‐2) in Natural Killer Cell (NK) Activation and Hypertension in a Preclinical Rat Model of Preeclampsia
Authors: Cunningham Mark W. , Ibrahim Tarek , Cornelius Denise , Amaral Lorena , Vaka Venkata Ramana , LaMarca Babbette
Published: June 2021

Women with preeclampsia (PE) have hypertension, placental ischemia, increased pro‐inflammatory cells (natural killer (NK) and Th1 cells) and cytokines, and intrauterine growth restriction (IUGR). Interleukin‐2 (IL‐2) is a Th1 cytokine that stimulates NK cell activation. We hypothesize that IL‐2 is one mechanism whereby Th1 cells, which are activated in response to placental ischemia, mediate NK cell activation, hypertension, and IUGR during pregnancy. The objective of this study was 2 fold: 1.) to determine if elevated IL‐2 during pregnancy will increase blood pressure, NK cell activation, and cause IUGR. 2.) to determine if blockade of IL‐2 (basiliximab) will decrease blood pressure, NK cell activation, and improve pup weight in the reduce uterine perfusion pressure (RUPP) preclinical rat model of PE The RUPP rat is a model of placental ischemia during pregnancy which has increased IL‐2 levels and mimics many of the symptoms of PE.

Methods

On gestational day 14, normal pregnant (NP) rats received IL‐2 infusion (0.20 ng/ml) via mini‐osmotic pump; while RUPP rats receive basiliximab (0.07 mg/per rat) via IV infusion. On day 19, conscious blood pressure (MAP), pups, placentas, and blood were collected. Flow cytometry was used for quantification of NK cells.

Results

IL‐2 infusion did not increase MAP (106 ± 3 vs. 100 ± 2 mmHg) nor change pup weight (2.24 ± 0.06 vs. 2.33 ± 0.06 g). However, IL‐2 infusion decreased placental weight (0.60 ± 0.02 vs. 0.67 ± 0.02 g, p<0.05 vs. NP) and increased circulating activated NK cells (10.4 ± 1.8 vs. 3.1 ± 1%, p<0.05 vs. NP). RUPPs rats exhibit elevated MAP (123 ± 3 vs. 103 ± 3 mmHg, p<0.05 vs. NP), placental (14.1 ± 2.9 vs. 7.8 ± 0.6%, p<0.05 vs. NP), and circulating (16.6 ± 1.3 vs. 7.7 ± 0.9%, p<0.05 vs. NP) activated NK cells. Basiliximab treatment had no effect to lower MAP (123 ± 2 vs. 123 ± 3 mmHg) or improve pup weight (2.08 ± 0.07 vs. 2.04 ± 0.07 g) and placental weight (0.59 ± 0.03 vs. 0.53 ±0.04 g) in RUPP rats. However, basiliximab decreased total placental NK cells in RUPP rats (4.6 ± 1.4 vs. 14.1 ± 2.9%, p<0.05 RUPP + basiliximab vs. RUPP).

Conclusion

IL‐2 appears to play a major role in NK cell activation during PE that could possibly impact placental health without causing increases in blood pressure during pregnancy.

Support or Funding Information

Research Supported by T32HL105324 and HL130456

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Association of Chronic Nicotine Inhalation with Hypertension in Mice
Authors: Fuchs Robert Marcus , Oakes Joshua , Basting Tyler , Lobell Thomas , Gilpin Nicholas , Gardner Jason , Yue Xinping , Lazartigues Eric
Published: June 2021

For decades it has been known that chronic tobacco smoke exposure is a risk factor for hypertension, predisposing to cardiovascular pathologies including stroke and heart failure. However, no prior study has evaluated the role of inhaled nicotine per se. Previous investigations found that subcutaneously‐administered nicotine acutely increases expression of angiotensin‐converting enzyme (ACE) and the angiotensin receptor (AT1R). These findings suggest RAS dysfunction may mediate nicotine‐induced hypertension through either increased expression of pro‐hypertensive proteins (ACE/AT1R) or suppressed expression of compensatory agents (ACE2/MasR).

Accordingly, we hypothesized that chronic nicotine inhalation promotes hypertension by inducing RAS dysfunction within autonomic brain nuclei. To test this hypothesis we exposed male (n = 64) and female (n = 44) mice to air (control) or twelve hours of nicotine vapor nightly for eight weeks. Mean nocturnal systolic blood pressure (telemetry) increased between the baseline and final week of nicotine exposure (males: +19.71 ±0.65 mmHg, F = 528.2, n = 5, p < 0.0001; females: +14.04 ±0.85 mmHg, F = 322.7, n = 4, p < 0.0001). In females, a trend towards elevated urinary norepinephrine was observed (t = 1.751, p = 0.0993). In a pilot study, when we exposed rats for 3 weeks to 14 hours of nicotine nightly, the hypothalamic ACE2 mRNA levels were reduced (t = 2.490, p < 0.05). In mice we found a trend towards increased expression of MasR mRNA in this brain region (t = 2.373, p = 0.0766).

Together, these findings suggest that nicotine inhalation contributes to high blood pressure, possibly through enhanced sympathetic activity. Mechanistically, our pilot data indicate that impaired ACE2 expression occurs in tandem with a compensatory increase in MasR in the hypothalamus of subjects inhaling nicotine. Future studies will elucidate activity of additional RAS contributors in cell culture and clarify the nature of autonomic dysfunction in vivo (e.g., increased sympathetic output vs decreased vagal tone). Collectively, this project will provide crucial information regarding the risk of hypertension in users of nicotine inhalation devices (i.e. e‐cigarettes), improving the ability of clinicians to treat this condition.

Support or Funding Information

This work was supported by NIH NHLBI grants 1R01 HL135635 and 2R01 HL093178.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Probiotic Administration in an Endometriosis Animal Model Can Influence the Gut Microbiota and Gut‐Brain Axis to Counteract the Effects of Stress
Authors: Chompre Gladys , Cruz Myrella L. , Arroyo Gerardo A. , Rivera Raquel M. , Colon Maria C. , Appleyard Caroline B.
Published: June 2021

Endometriosis is characterized by endometrial tissue growing in areas outside the reproductive system resulting in discomfort, pain, and infertility. Environmental factors such as stress have been suggested to exacerbate the condition, and our group has previously shown that stress can increase lesion size in a rat model. Animal studies have found increased gram‐negative bacteria in the gut microbiome of endometriosis animals and that oral administration of Lactobacillus gasseri can decrease lesion size however it is unclear whether probiotic use could counteract the effects of stress in our model.

Hypothesis

Administration of a probiotic mixture can decrease endometrial implants in animals under stress via modulation of the intestinal microflora.

Methods

Endometriosis was induced in female Sprague Dawley Rats (11–12/group) by implanting uterine tissue next to the intestinal mesentery on day 0. Animals were allowed to recover for seven days before administering placebo (Pl) or probiotic (Pro) in drinking water until time of sacrifice (day 60). Stressed (S) animals were subjected to water avoidance for 60 mins/day (days 14–20). Non stressed (NS) animals stayed in a clean cage. Anxiety was measured before and after stress using open field. Fecal bacterial composition was analyzed at four time points: before surgery, before stress, after stress, and at sacrifice.

Results

ESPl group had increased anxiety and development of more endometrial lesions (91.27%) which were of a larger size (38.31 ± 6.90mm2) compared with ENSPl (62.50%; 17.94 ± 3.90mm2, p<0.05). Administration of the probiotic (ESPro) decreased the number developed (56.25%), the size of the lesions (14.33 ± 3.17mm2, p<0.01), as well as colonic damage (p<0.05). Stress caused changes in the microflora including lower alpha diversity and altered firmicutes/bacteroidetes ratio, and these changes appeared to be reversed in those animals receiving the probiotic. Probiotic administration also modulated the effect of stress on bacterial abundance at the genus level.

Conclusions

Our results suggest that the gut microflora plays an important role in the stress induced‐exacerbation of the endometriosis symptoms in this animal model since administration of a probiotic mixture can decrease lesion size. This offers the potential for further exploration of this complementary therapy in the patient population.

Support or Funding Information

Supported in part by P20 GM103475‐15, R25GM082406, 2R25GM096955 & MD007579.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Fasting‐Mimicking Diet and Risk Factors for Aging, Diabetes, Cancer and Cardiovascular Disease
Authors: Shelechi Mahshid , Wei Min , Brandhorst Sebastian , Morgan Todd , Dorff Tanya , Hong kurt
Published: June 2021

Background

Prolonged fasting in which only water is consumed for 2 days or more has been shown to reduce pro‐growth signaling, activate cellular protection mechanisms and promote protection against cancer, diabetes, heart disease and neurodegenerative diseases. Since prolonged fasting is difficult for most human subjects, we developed a Fasting Mimicking Diet (FMD) which is a low calorie, low protein, plant‐based meal plan for 5 continuous days. During periods of low calorie intake, the body enters a protected mode, removes damaged cells and tissues, and undergoes self‐repair.

Methods

100 participants without a diagnosed medical condition in the preceding six months were enrolled based on Inclusion criteria (generally healthy adult volunteers, 18–70 years of age, body mass index: 18.5 and up). This study is designed as a randomized cross‐over trial, includes two arms: Control and multi‐cycle special 5‐day dietary regimen (Diet, 3 cycles). We evaluated the effects of the FMD on risk factors and markers for aging, cancer, metabolic syndrome and cardiovascular diseases.

Results

For all subjects who completed 3 FMD cycles (combining both FMD arms), body weight, BMI, total body fat, trunk fat, absolute lean body mass, waist circumference, IGF‐1, systolic and diastolic blood pressure, total cholesterol and LDL were significantly reduced (P<0.05) and relative lean body mass (p=0.0002) was increased.

Conclusions

FMD cycles are effective in improving an array of metabolic markers/risk factors associated with poor health and aging and in contributing to the treatment and prevention of multiple age‐related diseases.

Support or Funding Information

Funding was provided by the USC Edna Jones chair fund. Dr. Mack's contributions were provided through the Southern California Clinical and Translational Science Institute (SCCTSI) supported by NIH ULTR001855
A Mathematical Model for the Population Dynamics of Intestinal Microbiota: Obese Versus Healthy Gastrointestinal Tract
Authors: Arango Felipe Giron , Umaña Alejandro Pizano , Prieto Paola Calvachi , Cordovez Juan Manuel
Published: June 2021

Introduction

The term microbiota refers to a community of microorganisms that are located in a certain space that have a symbiotic relationship with the host cells, in this specific case the human gastrointestinal tract. This interaction provides support to the tract in different functional aspects such as trophic functions, protection (immune system) and metabolism. There has been evidenced that intestinal microbiota plays an important role in synthesis, metabolism, and other functions associated to the nutrients and micronutrients that can lead to healthy or obese patients. In this work we aimed to understand and model the population dynamics of the intestinal microbiota.

Methods

A mathematical model was design based on the gastrointestinal tract seen as a single tubular structure composed of Esophagus, Stomach, Small intestine (Duodenum, Jejunum and ileum) and colon. Only one major phylum of microorganisms (Either Firmicutes or Bacteroidetes) was included according to anatomical and physiological characteristics and in terms of its relative abundance in each part of the tract. The model included the subject's diet and activity level as inputs. Parameters for the model were obtained from a literature review. The model was implemented in Matlab and produced microorganism distribution (i.e. spatial concentration). We model different diets and pathological conditions.

Results

The model reveals an expected growth of either Firmicutes or Bacteriodetes associated with the amount of nutrient and carrying capacity of the gastrointestinal tract. These results are in agreement with several reports obtained from the literature. In addition, the model showed an interesting stabilization of bacterial population under some circumstances depending on the segment of the gastro intestinal tract. Bacterial population is sensitive to diet and exercise.

Conclusions

The model can be used to predict the population dynamics under different diets and furthermore could be implemented as an important tool for bariatric surgery patients. The model can also shed some light on how to prevent the progression from overweight to obese just by changing the diet understanding the dynamics of gastrointestinal tract population.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Computational Analysis of 1,2‐dialkynylpyrroles
Authors: Bondoc Joshua , Kerwin Sean
Published: June 2021

Computational chemistry can be used to quickly predict the properties of molecules. We are interested in studying the potential of 1,2‐dialkynylpyrroles to undergo a thermal Bergan cyclization‐triggered rearrangement to reactive diradical intermediates that display cytotoxic activity. These diradical intermediates have garnered much interest in the scientific community as a potential tumor suppressing compound. In order to analyze these diradicals, a B3LYP hybrid functional with a 6–31G** basis set was used to predict the electronic energy of each intermediate and transition state in the reaction. The electronic energy values for each step in the reaction were used to construct an energy diagram that quantified the energy gap between the intermediates and transition states. The goal of this experiment was to create an analogue of a dialkynylpyrrole that maximized the energy gap between the diradical singlet and the Retro Bergman transition state, and minimized the energy gap between the diradical singlet and the diradical triplet. An analogue that produced a diradical with these properties would be long lived and reactive.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification of XbaI polymorphism of the estrogen receptor alpha in mexicans at early posmenopause
Authors: Lugo‐Martínez Gabriela , Jiménez‐Zamarripa Carlos Alberto , Madrigal‐Santillán Eduardo Osiris , Calzada‐Mendoza Claudia Camelia
Published: June 2021

INTRODUCTION

Menopause is the cessation of menstrual cycle, is diagnosed after at least twelve consecutive months whitout bleeding, it occurs due to the reduced functioning of the ovaries, resulting in lower levels of ovarian hormones primarily estradiol (NAMS, 2012). Estrogen suppression has been correlated with higher incidence of cardiovascular and metabolic diseases observed in this age group.

Estrogens exert its actions through their receptors, the most studied isoforms of these receptors are: the alpha (ER‐α), beta (Er‐β) and the a G coupled (GPR30). Due to its action and location, an important role in the preservation of cardiovascular and metabolic health has been attributed to ER‐ α (Raushember y Cols, 2016).

Several ER‐α polymorphisms have been described, one of the most interesting due to its association with the etiology of cardiovascular pathologies, is the termed as XbaI (rs9340799A> G). This one, consists in the change of an adenine for a guanine, which modifies the affinity of the receptor for its ligand. The X allele has been linked to increased risk of bone demineralization, dyslipidemia, high body mass index (BMI), and oncologic diseases, such as breast cancer and ovarian tumors (chedrahui y Cols,2014; Lobo y Cols, 2014).

The aim of this study was to identify the genotype of XbaI polymorphism in mexican posmenopausal women.

MATERIALS AND METHODS

A descriptive, observational, prospective, transversal, cohort study was carried out in 87 early posmenopausal women who attend to the climacteric clinic of “Hospital general Ignacio Zaragoza”. Clinical data, hormonal profile, blood pressure, antropometric parameters were evaluated. XbaI polymorphism was identified by restriction fragment length polymorphism (RFPL) method.

RESULTS

The mean age was 53.01±5.08 years, the BMI average was 28.77±3.88 m/kg2. The averages of systolic, diastolic, and mean blood pressure were 116±10.44 mmHg, 78.7±9.39mmHg y 91.58±9.1 mmHg, respectively;27 patients had hypertension diagnostic under pharmacological treatment.

We found 58.87%, 29.89% and 19.24% for xx, Xx and XX genotypes respectively (Graph 1).

The averages of systolic, diastolic, mean blood pressure and BMI according XbaI genotype are showed in table 1, there was no statistical differences between groups.

CONCLUSION

The most common genotype in mexican posmenopausal women was xx.

There was no differences in blood pressure and BMI according genotype.

All patients had elevated values for BMI, independently of her XbaI genotypes

Support or Funding Information

none

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
EGR1 Interacts with TBX2 and Functions as a Tumor Suppressor in Rhabdomyosarcoma
Authors: Mohamad Trefa Salih , Davie Judith
Published: June 2021

Background

EGR1, one of the immediate‐early response genes, can function as a tumor suppressor gene or as an oncogene in cancer. The function of EGR1 has not been fully characterized in rhabdomyosarcoma (RMS), a pediatric cancer derived from the muscle linage.

Methods

EGR1 expression was characterized by RNA and protein analysis in RMS cells. EGR1 was ectopically expressed in RH30 cells by transfection of an EGR1 expression plasmid and selection of stable cell lines. A combination of techniques such as proliferation assays, scratch assays and soft agar assays were performed with RH30 cells expressing EGR1 to demonstrate the loss of oncogenic growth properties. Differentiation of RMS cells was detected by immunohistochemistry and RNA and protein analysis. Co‐immunoprecipitation studies were used to identify the interaction between EGR1 and TBX2 and gene expression changes were detected by RNA and protein analysis. TUNEL assays and RNA and protein analysis were used to characterize apoptosis in EGR1 expressing RH30 cells.

Results

We found that EGR1 is downregulated in the alveolar RMS (ARMS) subtype but expressed at levels comparable to normal skeletal muscle in embryonal RMS (ERMS). We found that over expression of EGR1 in ARMS significantly decreased cell proliferation, mobility, and anchorage‐independent growth. EGR1 also promoted differentiation in RMS cells by inducing the expression of several genes involved in muscle differentiation including myosin heavy chain (MHC), MYOD1 and MYOG. We also found that EGR1 interacts with TBX2, which we have shown functions as an oncogene in RMS. The interaction inhibits EGR1 dependent gene expression, which includes the cell cycle regulators CDKN1A and PTEN as well as other important cell growth drivers such as NDRG1 and CST6. We also found that EGR1 induces apoptosis by triggering the intrinsic apoptosis pathway. EGR1 also sensitized RMS cells to chemotherapeutic agents.

Conclusions

We show that EGR1 is a potent tumor suppressor in RMS and that EGR1 and TBX2 antagonize each other's function. Our data suggest that activation of EGR1 may improve the therapeutic targeting of RMS.

Support or Funding Information

Higher Committee for Education Development in Iraq (HCED)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Phosphoinositide Signaling Meets Heme Biochemistry
Authors: KHAN DANISH , Wofford Joshua , Gulten Gulcin , Sacchettini James , Lindahl Paul , Bankaitis Vytas
Published: June 2021

The Phosphatidylinositol Transfer Proteins (PITPs) have emerged as central regulators of phosphoinositide signaling that that diversify signaling outcomes in cells. Sec14 is the prototype PITP in S. cerevisiae with five homologs (Sfh1‐Sfh5) involved in activities as diverse as trans‐ Golgi signaling and lipid droplet metabolism to phosphatidylethanolamine and ergosterol metabolism. Sfh5 is an orphan member of this family with unknown function with limited information about its genetic interactions or chemogenomic signatures. The surface potential map of Sfh5 indicated unusual electrostatics in the lipid‐binding domain of the protein. We purified recombinant Sfh5 protein from E. coli and subjected it to rigorous biochemical and biophysical analysis. We show that Sfh5 is a heme binding protein with a high spin ferric center as characterized by EPR and Mössbauer Spectroscopy. We solved the crystal structure of the heme‐bound protein to 2.6Å resolution and show that the lipid binding and heme binding sites overlap. We also identified a novel heme‐coordination motif that is conserved through fungal Sfh5s. Finally, we show that the lipid transfer ability of Sfh5 is enhanced when the protein is stripped of heme. We speculate that this protein is a heme‐transfer protein that delivers hemes within cell, particularly under conditions of oxidative stress. Taken together, this is for the first time phosphoinositide signaling proteins have been linked with bioinorganic chemistry involving heme.

Support or Funding Information

This work was funded in part by a grant from the National Institutes of Health (GM44530) and by the Robert A. Welch Foundation (BE0017).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Imaging and Quantifying Mitochondrial Morphology: a Focus on the 3D Freeware MitoGraph
Authors: Harwig Megan Cleland , Viana Matheus P , Egner John M , Harwig Jason , Widlansky Michael E , Rafelski Susanne M , Hill R. Blake
Published: June 2021

Mitochondria are found in a variety of shapes, from small round punctate structures to a highly‐interconnected web. This morphological diversity is important for function, but complicates quantification. Consequently, early quantification efforts relied on various qualitative descriptors that understandably reduce the complexity of the network leading to challenges in consistency across the field. Recent application of state‐of‐the‐art computational tools have resulted in more quantitative approaches. This work focuses on MitoGraph, an open‐source image analysis platform for measuring mitochondrial morphology initially optimized for use with Saccharomyces cerevisiae. MitoGraph was used to assess mitochondrial morphology of five different mammalian cells types, all of which were accurately segmented by MitoGraph analysis. MitoGraph also successfully differentiated between distinct mitochondrial morphologies that ranged from entirely fragmented to hyper‐elongated.

Support or Funding Information

This was work was supported by the National Institutes of Health grants: R01‐GM067180, R01‐ HL128240 and R41‐AG056253.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
C‐terminal domain is responsible for a Kunitz‐type inhibitor uptake by tumor cells
Authors: Chudzinski‐Tavassi Ana Marisa , Morais Katia Luciano Pereira , Servent Denis , Sciani Juliana Mozer , Iqbal Asif
Published: June 2021

Amblyomin‐X is a recombinant Kunitz‐type FXa inhibitor that was identified through the transcriptome analysis of the salivary gland from Amblyomma sculptum tick. This protein triggers apoptosis in different tumor cell lines, and promotes regression of tumor growth and reduction of metastasis. In this regard, recently arose question about it structure‐fuction. Thereby, we investigated the effects of two truncated (synthetic) regions of Amblyomin‐X, which we termed Kunitz and C‐terminal domains. Here, using MTT and FXa chromogenic activity assay, we found that both domains have no cytotoxicity in human tumor cells (SK‐MEL‐ 28 and MIA PaCa‐2) and only Kunitz domain inhibit FXa slightly. Also, through confocal microscopy and high content analysis (ImageXpress Micro Confocal High‐Content, Molecular Devices), we observed C‐terminal domains (Alexa Fluor 488 conjugated) uptake by tumor cells previously than Amblyomin‐X (whole protein), but with similar intracellular trafficking and negligible nuclear translocation. Besides, circular dichroism analysis shown that Kunitz domain has quite a structure of α‐helix and β‐sheet, whereas the C‐terminal domain has the great part of random structure. Taken all results together, we can suggest that the C‐terminal domain is responsible for Amblyomin‐X uptake and could be an unusual cell penetrating peptide.

Support or Funding Information

Supported by BNDES, Fapesp and União Química Farmacêutica

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Translation of Physiological Understanding (PhUn) Week Activities to a Larger Multi‐ Level Group of Students in a Scientific Environment Following Hurricane Maria
Authors: Rodriguez‐Flores Christine D. , Cruz Myrella , Gonzalez‐Charles Agnes P. , Brofen‐Quiñones Samuel , Rivera Raquel , Colon Maria , Ramos Dihna , Appleyard Caroline B. , Chompre Gladys
Published: June 2021

Our previous PhUn week activities focused on students from similar grade levels in schools always achieved good learning outcomes. This year due to damaged school facilities after hurricane Maria devastated our island of Puerto Rico, we decided to introduce a Neuroscience theme in a larger combined activity held in a university laboratory environment.

Hypothesis

Having a larger number of students from different grade levels in a more scientific environment will promote learning and motivation.

Methods

Students from elementary (n=36), middle (n=33) and high school (n=33) attended an activity held at the local university. The activity was divided into three phases: a pre‐test, a teaching/interactive phase which included two conferences followed by rotating hands‐on activities, and a post‐test.

Results

As in previous years, all students learned new information (elementary, middle school, and high school (39%, 5% and 11% increase vs. pre‐test). However, in contrast to previous years, students asked many questions related to research and laboratory safety, and wanted to know more about how they could work in that type of setting.

Conclusions

Even in this larger group setting, all students gained knowledge about the nervous system, and the new environment produced higher levels of curiosity and excitement. Students were able to learn first‐hand what a physiologist is and what a career in this field might be.

Support or Funding Information

Supported by PUCPR

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Induction of inward arterial remodeling is ameliorated in vivo by inhibition of actin polymerization dynamics in a mouse model of hypertension
Authors: Foote Christopher A , Castorena‐Gonzalez Jorge A , Padilla Jaume , Martinez‐Lemus Luis A
Published: June 2021

Hypertension is associated with structural changes to the resistance vasculature leading to a reduction in resistance artery diameter. This process is termed inward remodeling and is defined as a reduction in lumenal diameter under passive conditions. We posit that inward remodeling involves dynamic changes to the actin cytoskeleton that facilitate the movement and rearrangement of vascular smooth muscle cells around a smaller vascular lumen. Ex vivo, prolonged exposure to vasoconstrictors has been demonstrated to induce remodeling via g‐protein coupled receptors (GPCR) that activate the Rho signaling pathway and subsequently promote actin polymerization. We report here that vascular smooth muscle cells, propagated on nanopatterned culture‐ware to maintain their contractile phenotype, increase their ratio of filamentous (F) to globular (G)‐actin in response to GPCR activation by Norepinephrine and Angiotensin (Ang) II. One of the enzymes in the Rho signaling pathway implicated in the increase in F/G‐actin following GPCR activation is Lim Kinase, which phosphorylates and inactivates the actin severing enzyme, cofilin. In isolated rat cremasteric arteries we demonstrate that inhibition of Lim Kinase blocks inward remodeling in the presence of GPCR agonists and this is coincident with a reduction in cofilin phosphorylation. To examine the process in vivo, we utilized a transgenic mouse line that expresses smooth muscle actin (SMA)‐GFP in vascular smooth muscle cells to measure proximal arterial diameter noninvasively within the outer ears using confocal microscopy. Exposure of these mice to 1000 ng/kg/min of Ang II for two weeks resulted in a significant increase in mean arterial blood pressure. Both ears were imaged prior to Ang II infusion (Day 0) and after 14 days of infusion to assess inward remodeling. Starting at day 0, the right ear was injected with a bolus of a Lim Kinase inhibitor (10 uM) and the left ear was injected with vehicle. Injections were repeated every 4 days for a total of four injections. The vehicle exposed ears significantly inwardly remodeled in contrast to those exposed to the Lim Kinase inhibitor (Vehicle diameter = −15.95 % +/− 1.79 versus Lim Kinase inhibitor diameter = −1.75% +/− 1.78, P<0.05). These data demonstrate that in vivo, Lim Kinase plays a necessary role in inward remodeling, and thus merits additional study as a potential therapeutic target in cardiovascular pathologies associated with inward remodeling.

Support or Funding Information

NIH‐HL‐088105 to LAM

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Impaired Coronary Collateral Growth in a Mouse Model of Diabetes
Authors: Cumpston Devan , Wan Weiguo , Chinchilla Sofia , McCabe Caige , Enrick Molly , Shoenbeck Sydney , Chilian William , Yin Liya
Published: June 2021

Cardiovascular disease is the leading cause of death in the United States, Ischemic Heart Disease (IHD) accounts for one in seven cardiovascular related deaths. Patients with poorly developed coronary collateral networks have a poorer prognosis after a cardiovascular event than those with well‐developed collaterals. Furthermore, coronary collateral growth is impaired in metabolic syndromes such as diabetes, and obesity. The underlying mechanism(s) that is (are) causal for growth are unknown. In an attempt to elucidate a mechanistic understanding, we developed a murine model that allows us to use genetically modified mice to study the coronary collateral growth spatially and temporally.

Methods

a pneumatic snare was implanted and situated around the LAD (open chest surgery during anesthesia). After mice recovered from surgery, the snare was periodically inflated to occlude the LAD; thus producing episodes of ischemia or repetitive ischemia (RI). Coronary collateral growth (CCG) was measured by myocardial blood flow with contrast echocardiography and by collateral numbers by micro‐CT.

Results

1) No native coronary collaterals were found in mice. 2) After the RI stimulation, extensive coronary collaterals grew. 3) Diabetic mice failed to grow coronary collaterals after RI stimulation. 4) Moreover, the coronary blood flow increased in the ischemic area when there was coronary collateral growth. 5) The quantification of vessel density by tree analysis showed an increase after RI stimulation. Figure 1 (A–C) shows the microfil pictures of the wild type mouse heart without CCG (A), the wild type mouse heart with CCG (B), and the diabetic heart without CCG (C). We believe this murine model of coronary collateral growth is very useful for future studies to investigate the mechanism of regulation of coronary collateral growth. Figure 1 (D–F) shows the micro‐CT pictures of the wild type mouse heart without CCG (D), the wild type mouse heart with CCG (E) and the diabetic heart without CCG (F).

Support or Funding Information

2R01HL103227‐05( Zhang, Yin), 1R01HL135110‐01 (WMC, LY), 1 R01 HL137008‐01A1 (LY), 1R15HL115540‐01 (LY) and14BGIA18770028 from American Heart Association

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.