bluuebunny/crossref_metadata_2025 huggingface dataset:
Lipids as potential contributors to the beneficial effects of Roux‐en‐Y gastric bypass surgery (RYGB)
Authors: Kaufman Sharon , Krieger Jean‐Philippe , Langhans Wolfgang , Arnold Myrtha
Published: June 2021

RYGB is still the most effective treatment of obesity and its comorbidities, but the signaling molecules and mechanisms that mediate the beneficial effects of RYGB are largely unknown. One of the classes of molecules that could be involved are gut‐derived endogenous lipids. We therefore investigated profiles of potential lipid mediators locally in the gut. We non‐occlusively cannulated the mesenteric lymph duct of diet‐induced obese (DIO), insulin‐resistant RYGB or sham‐operated rats and collected lymph samples in relation to spontaneous meals for subsequent mass spectrometry analysis of the intestinal lymphatic lipidome from all animals prior to and 5, 10 and 21 days after the surgery. A multivariate analysis revealed two separate clusters for the lipidomes of the animals of the two surgical groups. Unlike sham‐surgery, RYGB surgery changed the levels of >150 lipids compared to pre‐surgical levels. TG were mostly decreased after RYGB, presumably reflecting the reduced food intake. Some lipids, however, were increased in intestinal lymph after RYGB. Especially those reported to be decreased in obesity (e.g., lysophosphatidylcholine [LPC] 20:1 or phosphatidylcholine [PC] 32:0) were upregulated about two‐fold 21 days after the RYGB surgery. This might be related to the decreased body weight rather than being a direct surgery effect. Some lipids, however, (e.g., PC 40:7, phosphatidylethanolamines [PE] 40:6, sphingomyelins [SM] 18:0) were upregulated in the RYGB group already 5 days after surgery, long before their body weight was significantly decreased compared to sham‐operated controls. Notably, there was a strong positive correlation between RYGB surgery and longer chain length in particular for TG, diacylglycerols [DG], PE, and SM. Moreover, TG, DG and phosphatidylinositols showed a positive correlation between RYGB surgery and an increased frequency of double bonds. Whether these changes are cause or consequence of an improvement of enterocyte structure and function, and whether they contribute to the reported improvement of the various comorbidities requires further investigation.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exposure to a Western‐Style Diet during Early Development Reduces Skeletal Muscle Lipid Metabolism and CI‐dependent Oxidative Capacity in Juvenile Non‐Human Primate Offspring
Authors: Greyslak Keenan T , Hetrick Byron , Takahashi Diana A , Dean Tyler , Kievit Paul , Sullivan Elinor L , McCurdy Carrie E
Published: June 2021

Maternal obesity and diabetes during pregnancy is linked to an increased and earlier risk for offspring to develop insulin resistance and metabolic diseases. We have previously found in a non‐human primate model that maternal western‐style diet (WSD) alone or in combination with obesity reduces oxidative capacity in fetal muscle concomitant with increased oxidative damage and lipid metabolism. The purpose of this study was to determine whether fetal adaptations to maternal WSD alone independent of maternal obesity leads to persistent reductions in skeletal muscle metabolism in juvenile offspring prior to the development of obesity.

Experimental Design

Lean adult female Japanese macaques were maintained on a control diet (CTR) or WSD prior to and during pregnancy and lactation. Male and female offspring were than weaned to independent housing at 7 mo and fed a WSD or CTR resulting in four groups (CTR/CTR, CTR/WSD, WSD/CTR and WSD/WSD). Insulin sensitivity was measured by i.v. GTT and body composition by DEXA at 36 mo. Muscle biopsies were taken at 40 mo to measure oxidative metabolism by high resolution respirometry and electron transport system complex (C) activities by spectrophotometry. Data were analyzed by a 2‐way ANOVA (maternal diet X offspring diet)

Results

There were no differences in body weight, % fat or insulin sensitivity across groups. Total oxidative capacity and CI‐dependent oxidative capacity were decreased by ~30% (P=0.001) in the presence of lipid substrates in gastrocnemius (gastroc) and soleus of offspring exposed to a maternal WSD independent of offspring diet. CI‐specific activity was reduced by 50% (P<0.0001) in the gastroc. Citrate synthase activity was not different suggesting that reduced oxidative metabolism is not caused by diminished mitochondrial abundance.

Conclusions

Maternal WSD during pregnancy and lactation results in a persistent reduction in skeletal muscle lipid metabolism in offspring prior to offspring obesity. Switching offspring to a heathy diet did not ameliorate the effects of developmental exposure to maternal WSD. Impaired muscle lipid metabolism is linked to insulin resistance in adults and likely contributes to the increased risk of metabolic disease in exposed offspring.

Support or Funding Information

R24 DK090964R01 MH107508R01 (ES)

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effect of Ginger Powder Extracts on Actin Assembly In Vitro
Authors: Sugimoto Naotoshi , Katakura Masanori , Matsuzaki Kentaro , Miyamoto Mayumi , Sumiyoshi Eri , Yachie Akihiro , Shido Osamu
Published: June 2021

Objective

The components of ginger root (Zingiber officinale Roscoe) have been widely used for various medicinal purposes in Asia. Several bioactive compounds have been identified in ginger, including 6‐ and 8‐gingerols and 6‐shogaol, which are agonists of the capsaicin receptor TRPV1[1, 2]. Recently, we demonstrated that ginger powder might affect human metabolism in vivo[3]. However, the effects of the bioactive compounds of ginger on live cells have not been completely elucidated. In this study, we investigated whether ginger powder extracts (GPE) induced signal transduction, especially actin assembly, in vitro.

Method

The active components of GPE were characterized by high‐performance liquid chromatography. NIH3T3 mouse fibroblast cells were maintained in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum at 37°C in a 5% CO2 incubator. These cells were treated with GPE (10 ng/mL) for 10 min, followed by analysis of the activation of mitogen‐activated protein kinases (MAPKs) ERK and p38 MAPK, and actin assembly.

Results

The levels of phosphorylated ERK and p38 MAPK increased significantly after a 10‐ min stimulation with GPE. In addition, lamellipodia formation, which is known to facilitate cell migration[4], occurred at cell edges after GPE stimulation.

Conclusions

GPE activated MAPKs and increased lamellipodia formation in mouse fibroblasts. These results suggest that GPE modifies signal transduction, including actin assembly, in vitro.

Support or Funding Information

KAKENHI, Japan

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
H2S‐Induced Structural Changes of Insulin Protein in Early Lag Phase: Key to Detaining Amyloid Formation
Authors: Colon‐Rios Daniel Andres , Aldarondo‐Torres Alani , Rosario‐Alomar Manuel F. , Lopez‐Garriga Juan
Published: June 2021

Amyloid fibrils are formed by soluble proteins that assemble into insoluble fibers associated to many types of diseases such as Diabetes Type 2, and Alzheimer's Disease. Amyloid formation can be divided into three phases: the lag phase, where native protein unfolds and begins to self‐aggregate; the elongation phase, in which these aggregates grow into fibrils, and the steady phase, where saturation of mature fibrils occurs. Hen egg white lysozyme (HEWL) amyloid fibrils are inhibited in the presence of hydrogen sulfide (H2S) (Rosario, 2014). To further explain the means by which H2S inhibits amyloid fibrillation, insulin from bovine pancreas (IBP), was tested upon experimentation with increasing concentrations of H2S. ThT Fluorescence analysis revealed a decrease in peak intensity as the concentration of H2S increased in each sample, proving H2S to be an inhibitor of insulin amyloids. Atomic Force Microscopy (AFM) of the control sample showed an array of fibrillary structures. However, as the concentration of H2S increased, samples went from having a combination of fibrillary structures and small conglomerated spherules to the sole appearance of small spherules. Far UV‐Circular Dichroism spectra of the native protein and the protein interacting with H2S were very similar. However, non‐resonance Raman spectra revealed the presence of newly formed trisulfide bridges in the interaction between insulin and the H2S species as opposed to the disulfide bridges found in native and fibrillated insulin protein. The data suggest H2S‐protein interactions favor a structural change in the process of protein denaturing. The H2S species intervenes with amyloid formation early on during the lag phase inhibiting the formation of protofibrils and potential mature fibrils.

Support or Funding Information

Research reported in this publication was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103475. It was also supported by the National Institute of General Medical Sciences of the National Institutes of Health under Award Number T34GM008419. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Protective effects of Enterococcus faecium strain R30 on transformation of slow to fast fiber under inactive condition
Authors: Ikeji Takuya , Hirayama Yusuke , Matsumoto Tomohiro , Ono Kohei , Takuwa Miho , Hirabayashi Takumi , Tategaki Airo , Kondo Hiroyo , Fujino Hidemi
Published: June 2021

Inactivity results in the transformation of slow to fast fiber and mitochondria dysfunction in skeletal muscle. PGC‐1α, a transcriptional factor, can prevent transformation of slow to fast fiber and the reduced oxidative enzyme activity in skeletal muscle. We have confirmed that Enterococcus faecium strain R30 (R30) activated muscle sympathetic nerve and increased blood flow in muscle capillary in our previous study. Several studies report that the increased blood flow induces nitric oxide synthase (eNOS) expression via shear stress, and eNOS expression regulates PGC‐1α protein expression. Also, the activated sympathetic nerve activity increased PGC‐1α protein expression via activation of adrenergic receptor. Therefore, we examined the hypothesis that R30 might prevent the transformation of slow to fast fiber and the reduced oxidative enzyme activity via PGC‐1α protein expression in skeletal muscle under inactive condition. Fifteen male Sprague‐Dawley rats aged 11 weeks old were divided into three groups, that is, 1) control (CON; n = 5), 2) 2 weeks hindlimb unloading (HU; n = 5), and 3) HU+R30 (HU+R30; n = 5). In the experimental periods, we administered orally to R30 (500 mg/kg/day) in the HU+R30 group, and the same amount of physiological saline in the CON and HU groups. After 2 weeks hindlimb unloading, type I fiber composition ratio, citrate synthase (CS) activity, eNOS and PGC‐1α protein expression level were measured. Type I fiber composition ratio and CS activity were significantly higher in the HU+R30 than in the HU group. The expression levels of eNOS and PGC‐1α protein in the HU+R30 group were significantly higher than in the HU group. These results indicate that R30 supplementation could increase the levels of eNOS and PGC‐1α expression in soleus muscle, and prevent the transformation of slow to fast fiber and the reduced oxidative enzyme activity under inactive condition.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Growth Arrest Specific 6 in Regeneration after Skeletal Muscle Injury
Authors: Tsukamoto Jacob J , Matsumura Marc S , Olsen Zachary E , Wilson Esther K , Chow Joshua L , Arroyo Juan A , Thomson David M
Published: June 2021

Growth arrest‐specific 6 (Gas6) belongs to the family of vitamin K‐dependent proteins. Its functions in the cardiovascular, pulmonary and immune systems have been reported on in depth, but very little is known about its role in skeletal muscle. Because of its roles in the regulation of inflammation and cell proliferation in other tissues, our objective was to determine whether Gas6 plays an important role in skeletal muscle repair and/or regeneration after injury.

Therefore, we unilaterally injured the tibialis anterior (TA) muscles of young adult (YA; 5 mo.) and old (O; 30 mo.) Fisher Brown‐Norway rats with barium chloride (n=5–8/group). At the time of injury we also implanted a subcutaneous osmotic pump that continuously infused Gas6 (4 mg/ml) or saline onto the TA muscle through silicone tubing. The contralateral muscles served as controls. One week after injury, blood and the TA muscles were collected and the rats were euthanized. Muscle sections were prepared and stained with hematoxylin and eosin. Muscle were homogenized and Gas6 concentration was measured by ELISA in both plasma and muscle homogenates.

We found that muscle injury greatly increased Gas6 concentration in muscle, but not plasma. [Gas6] was greater in O vs. YA plasma, but age had no effect on Gas6 levels in skeletal muscle. Gas6 infusion increased regenerating muscle fiber size in YA rats, but not in O rats (n=3), despite a similar increase in Gas6 concentration with injury.

These results suggest that Gas6 is likely an important player in the regeneration of skeletal muscle after injury, and that the impaired recovery of old skeletal muscle after injury may be due in part to Gas6 resistance.

Support or Funding Information

Research Support: BYU Gerontology Program Grant

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Exercise, but Not Metformin Prevents Muscle Function Loss due to Doxorubicin in Mice Using an In‐Situ Model
Authors: Mackay Amy D , Hancock Chad R
Published: June 2021

Doxorubicin (DOX) is a clinically relevant chemotherapeutic known to cause severe side effects in heart and skeletal muscle. The deleterious effects DOX elicits on skeletal muscle are concerning due to the high rates of severe fatigue experienced by chemotherapy patients. Identifying treatments that prevent this skeletal muscle dysfunction is critically important. Exercise (EX) and metformin (MET) have both been suggested as treatments to reduce the toxic effects of DOX. However, their effects on muscle function in the gastrocnemius‐plantaris‐soleus (GPS) complex are unknown. Additionally, the mechanism by which DOX decreases muscle function are still under investigation. DOX can cause mitochondrial disruption, but the direct effects of mitochondrial function on muscle function have not been studied in this model. The purpose of this study is three fold: (1) to determine if DOX decreased muscle function of the GPS complex using an in‐situ muscle fatigue protocol, (2) determine if EX or MET treatment can prevent muscle function loss due to DOX, and (3) determine if muscle function loss is due to a decrease in mitochondrial respiratory function. C57BL/6 mice were randomly assigned to 4 groups: CON, DOX, DOX+EX, or DOX+MET. DOX was administered as a single intraperitoneal injection at 15 mg/kg body weight 3 days before sacrifice. Moderate EX treatment was done by treadmill running for 60 minutes for 5 days at 70% max prior to DOX injection. MET treatment was administered via oral gavage at 500 mg/kg body weight for 5 days beginning 2 days before DOX injection. GPS complex muscle function was measured in‐situ using tetanic contractions elicited by direct stimulation in a 6 minute fatigue protocol. Real time mitochondrial respiratory function was measured in gastrocnemius muscle. We report that DOX treatment caused ~20% body weight loss at 3 days, an effect that was partially prevented in DOX+EX (p<0.05) or DOX+MET (p=0.06). A small decrease in total force production was observed with DOX, which trended towards significance (0.08≥p≤0.04). DOX+EX treatment restored force production back to CON levels, but DOX+MET did not. The half relaxation rate was impaired by an average of 18% with DOX treatment (p<0.05). DOX+EX prevented this impairment of the half relaxation rate (p<0.05). DOX+MET did not restore impaired half relaxation rate caused by DOX. Mitochondrial respiration was not different between the groups, suggesting that the changes in muscle function occur independent of mitochondrial function. In conclusion, DOX treatment decreased muscle function in the GPS complex by impairing half relaxation rates. EX but not MET treatment prevented impaired half relaxation rate. These changes occurred independent of mitochondrial respiration.

Support or Funding Information

Mentoring Environment Grant, Brigham Young University

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
An anatomical study to determine the safest posterior arthroscopic approach to the human hip
Authors: Curcio Daniella F , Gomes Berliet Assad , Ramos Max Rogerio Freitas , de Almeida Camila Rodrigues , Fiorelli Stenio Carlos Alvim , Fiorelli Rossano Kepler Alvim , Laitman Jeffrey T.
Published: June 2021

The human hip (acetabulofemoral joint) is amongst the most complex joints for arthroscopic surgery. This is due to the many inviolable structures that are in direct contact, proximity, or surround the hip joint, notably: the femoral neurovascular bundle, anteriorly; the lateral cutaneous nerve of the thigh, in the anterolateral aspect; and gluteal vessels and the sciatic nerve (SN) in the posterior region. Arthroscopic access of the hip joint involves the opening of cutaneous entry portals. We here discuss the posterior portal (PP), placed at 1cm proximal and 1cm posterior to the palpable apex of the great trochanter, where its anterior and posterior edges merge superiorly, in neutral position or slight interior rotation of the hip (Aprato et al. Muscles, Ligaments and Tendons Journal. 2016). The PP offers a good view of the posterior aspect of the articular capsule of hip, but involves risk of injury to the SN and anatomical variations should be considered. This study evaluates the types of anatomic variations of the SN and assesses the risk of nerve injury by measuring the distance between the nerve and the PP in 40 cadaveric hip joints (17 males and 3 females). We used a millimeter caliper to obtained the measurements and the standard Beaton's and Anson's criteria (Anat Rec. 1937) as a reference to classify the anatomic variations. From our sample, 35 specimens presented with variation type I: SN emerging below the piriformis muscle (PM) in a single bundle; and 5 specimens presented with variation type II, common peroneal nerve running through the PM and the tibial nerve emerging below the PM. The average distance from the sciatic nerve to the PP was 3.06cm for type I variation group, 2.46cm for the type II variation group (p<0.05, Wilcoxon and t test). In the cases presenting with type II variation, the SN is closer to the region of access via PP and the risk of injury is higher. We recommend a careful analysis of the MRI, prior to the surgery, with attention to the anatomical variations of the SN, and a maximum distance of 2.0cm from the cutaneous PP in order to increase the surgical safety of posterior arthroscopic approaches of the human hip joint.

Support or Funding Information

In conjunction with the Building Bridges initiatives

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Morphological Variations of The Ovine Heart Interatrial Septum
Authors: Holda Mateusz Krystian , Holda Jakub , Tyrak Kamil , Klimek‐Piotrowska Wieslawa
Published: June 2021

Background

Atrial septal pouch (SP) is defined as a blind ended, kangaroo‐like diverticulum that may be located on the left or/and right side of the interatrial septum of the heart. A left‐sided SP may trigger atrial fibrillation and be a source of thrombosis, which increases the risk of ischemic stroke. Recent discoveries on the clinical significance of left‐sided SP may encourage the creation of new strategies and devices for the management of SPs. However, these strategies may first need to be tested in the ovine model before implementation in humans. Unfortunately, little is known about the presence of SPs in this species. Thus, we evaluated the morphology of the interatrial septum in ovine hearts and compared them with that of human and swine.

Methods

In this study a total of 60 ovine (Ovis aries) hearts were examined. The interatrial septum morphology was assessed as well as the patent foramen ovale (PFO) channel and SPs were measured. This study has been conducted in conformance with the FASEB Statement of Principles for the use of Animals in Research and Education and the Basic Principles of the Declaration of Helsinki and their expansion.

Results

The most commonly occurring variant were PFO channels (25.0%) with channel lengths of 5.4 ± 2.3 mm. Smooth septums were observed in 18.3% of hearts. In the remaining cases, septums had a left septal ridge (15.0%), left SP (11.7%), left septal bridge (10.0%), right SP (10.0%), or had both a right SP and left septal ridge (10.0%). No double SPs were observed. The mean right SP depth was 3.4 ± 1.2 mm, and its mean ostium width and height were 7.9 ± 1.8 mm and 2.8 ± 1.0, respectively. For the left SP, the mean depth was 6.0 ± 1.7 mm, the ostium width was 7.9 ± 2.4 mm, and the ostium height was 4.1 ± 1.6 mm (range: 2.3 to 6.4 mm). The left SPs were significantly deeper than right SPs (p=0.001) and had larger ostium height (p=0.04). There were no differences between the dimensions of the right SPs when the left septal ridge was present or absent (p >0.05). There were no correlations between any measured SPs dimensions and heart weight (p >0.05).

Conclusions

In conclusion the interatrial septum of ovine hearts exhibit morphologies that are more similar to humans than they are to swine. The presence of a left SP in sheep hearts make ovine models a promising alternative to the human heart for developing left‐sided SP management devices and techniques.

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 Activity of LHx2‐Associated cis Regulatory Modules
Authors: Mulder Cassidy G , Watson Billy A. , Pira Charmaine U. , Shin Alexandra , Oberg Kerby C.
Published: June 2021

Limb outgrowth is controlled, in part, by fibroblast growth factor (FGF) and sonic hedgehog (Shh). FGF up‐regulation of SHH is essential for coordinated limb pattering; however, not much is known about this mechanism. We recently found that LHX2 is an intermediate in FGF‐mediated SHH up‐regulation. LHX2 is part of a family of genes that are known to integrate signaling events. Thus, we hypothesized that FGF would up‐regulate LHX2 through associated potential cis regulatory modules (PCRM). Multiple PCRMs were found in association with the LHX2 locus. PCRMs were screened using comparative genomics, proximity to the LHX2 promoter, and transcription factor binding sites related to FGF signaling. To exam the role of suspected PCRMs, we electroporated PCRM‐reporter constructs into chick embryos. We found one PCRM with activity in the brain coincident with LHX2 expression, but none of our initial PCRMs exhibited activity in the limbs. We reevaluated the LHX2‐associated PCRMs in light of limb‐specific chromatin activation marks and identified 5 additional PCRMs for analysis. Studies are underway to determine limb‐related PCRM activity of this new cohort. Investigations regarding the relationship between FGF and LHX2 will provide important insights into the mechanisms underlying the FGF‐SHH regulatory loop.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Differences in the Serum's Metabolomics in Patients with High Plasma Atherogenic Index and Healthy Controls. Preliminary Results
Authors: Gordillo‐Moscoso Antonio , Almendra‐Pegueros Rafael , Valencia Liliana , Esmer‐Sánchez David , Florez‐Ramírez Rogelio , Rodrigez‐Aguilar Maribel , Correa‐González Jose Antonio , Tejerina Teresa , Pierdant‐Perez Mauricio , Medina Ursula
Published: June 2021

Obesity is ones of the majors public health problems around the world, according to the World Health Organization, there were 1.9 billions adults with overweight or obesity. As we know, weight gain is associated with an increase in cardiovascular risk. The Atherogenic Index of Plasma (AIP) is the log transformation of the ratio form triglycerides between HDL cholesterol in serum. This ratio is a good measurement for the cardiovascular risk, when result >0.5 the risk for cardiovascular diseases is high. Until now metabolomics is a good tool to identify previous risk to any diseases, like gastrointestinal, immunological and cardiovascular. The aim was to compare the serum's metabolomics in patients with high plasma atherogenic index and healthy controls. We designed a case‐control study with the participation of 35 patients (12 cases and 23 controls), who complete the selection criteria (both sex, 18–75 years old, and without other diseases) and signed informed consent duly explained approved by the ethics committee. We collected anthropometric data as: weight, height and body composition; and biochemical data: HDL, LDL, total cholesterol, and triglycerides, for the calculation of AIP. For establish the differences in the serum's metabolomics and identify the most important variables of the group we did a Principal Component Analysis (PCA), random forest and Partial Least Squares (PLS) in R statics software. Results and Conclusion: The cases showed more waist circumference, triglycerides, HDL, total cholesterol, and IAP, these differences were statistically significant. (Table 1). We identify 5 principal components (56.3% variance explicated) the more representative metabolites in the cases were: 1‐Hexane,3–4 dimethyl and Heptane,2,3, dimethyl. For the controls, Cyclopentane methyl and 2H‐Pyran, Tetrahydro, more studies are necessaries for confirms these findings.

Support or Funding Information

The authors acknowledge grants from National Council for Science and Technology—National Problems (# 2015‐1340) and Infrastructure‐CONACYT (#2015‐253004).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Iron supplementation modulates genes involved in oxidoreductase activity, including prostaglandin‐endoperoxide synthase 2 in intestinal tumors
Authors: Swain James , Li Chunbiao , Leahy Patrick
Published: June 2021

To determine the effect of excess dietary iron on gene expression of intestinal tumors in an animal model of human familial adenomatous polyposis, male C57BL/6J ApcMin/+ mice were fed adequate and excess amounts of iron (ferrous sulfate: 45 vs. 450 mg iron/kg AIN‐93M[M] diet). After 10 wk, mice were sacrificed and intestines removed. Tumors and adjacent non‐tumor tissue were excised and RNA isolated for micro‐array analysis (n=3/group; Mouse Genome 430A 2.0 arrays; Affymetrix) and RT‐PCR. Pathway mapping was performed to reveal gene networks (Pathway Studio 4.0, Ariadne Genomics, Inc.; GeneSpring, Agilent, Inc.; Searcher, Lucidyx). Bayesian ANOVA was used for micro‐arrays. Data show that intestinal tumors alter expression of a group of genes involved in oxidoreductase activity when exposed to excess iron. Differentially expressed genes include prostaglandin‐endoperoxide synthase 2, retinol saturase, alcohol dehydrogenase, deiodinase, cytochrome P450 (family 2, 4), and glutathione peroxidase. Findings reveal that iron supplementation modulates oxidoreductase activity of intestinal tumors, possibly in response to the pro‐oxidative, pro‐inflammatory effects of excess intraluminal iron.

Support or Funding Information

Supported in part by grants IRG‐91‐022 from ACS (to JS) and P30CA43703 from NIH. Ferrous sulfate provided by Crown Technologies, Inc.

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 effect of congenic myostatin inhibition on a mouse model of osteogenesis imperfecta (OI) type I/IV
Authors: Aguillard Ashley Michelle , Gremminger Victoria L , Daghlas Salah A , Jeong Youngjae , Lenz Kristin L , Phillips Charlotte L
Published: June 2021

Bone is a dynamic organ in part due to its mechanosensitivity. Muscle mass and force are some of the largest physiological loads that bone experience during its life time, with muscle mass being directly proportional to bone mass. One of the factors regulating muscle mass is myostatin, a member of the TGF‐β superfamily, that acts as a negative regulator of muscle growth. Osteogenesis imperfecta (OI), commonly known as brittle bone disease, is a heritable connective tissue disorder characterized by skeletal fragility. A glycine to cysteine substitution in the proα2(I) collagen chain leads to reduced bone mineral density and compromised biomechanical integrity as modeled by the heterozygous G610C (+/G610C) mouse. +/G610C mice model both the genotype and phenotype of 64 affected individuals with mild to moderate type I/IV OI in an Old Order Amish kindred. We investigated the effects of myostatin deficiency on the +/G610C mouse muscle by breeding heterozygote myostatin deficient mice (+/mstn) with +/G610C mice to produce WT, +/mstn, +/G610C, and +/mstn +/G610C mice. After weaning at three weeks of age, mice were weighed weekly until 4 months of age when they were euthanized. After euthanization, 5 hindlimb muscles, the gastrocnemius, quadriceps, tibalis anterior, plantaris, and soleus, were harvested and weighed. Serum was separated from the blood and used to quantify myostatin levels. Serums from +/mstn and +/mstn +/G610C mice demonstrated decreased levels of myostatin as compared to WT and +/G610C mice. +/mstn mice showed increased body weights at four months of age when compared to WT littermates. +/mstn +/G610C mice also had increased body weights as compared to +/G610C littermates, although they did not reach significance. More importantly, however, +/mstn and +/mstn +/G610C mice exhibited significantly increased hindlimb skeletal muscle weights and relative muscle weights as compared to +/G610C mice for the gastrocnemius and quadriceps. These findings demonstrate that myostatin deficiency can lead to increased muscle mass in mice with OI which can potentially lead to increased bone mass and strength.

Support or Funding Information

Supported by March of Dimes, National Institutes of Health/National Institute of Arthritis and Musculoskeletal and Skin Diseases (RO1 AR055907), the Leda J. Sears Trust, the Kansas City Area Life Sciences Institute Patton Trust Research, Inc., and the University of Missouri Research Board Grant. The project described was also supported by the IMSD EXPRESS Fellows Program (or simply the IMSD EXPRESS Program) via grant number R25GM056901 from the National Institute of General Medical Science (NIGMS), a component of the National Institutes of Health (NIH).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Impact of Transient ACE Inhibition on Cardiac Reactive Oxygen Species Enzyme Expression in Response to Nitric Oxide Synthase Inhibition
Authors: Khokar Bilal , Garvin Alexandra , Perez Octavio , Hale Taben
Published: June 2021

We have previously shown that transient treatment with the angiotensin converting enzyme (ACE) inhibitor, enalapril, protects against future nitric oxide synthase (NOS) inhibitor, Nw‐Nitro‐L‐arginine methyl‐ester (L‐NAME)‐induced pathological cardiac remodeling. This protection is characterized by reduced inflammation, fibrosis, and collagen deposition. ACE inhibitors have been shown to reduce oxidative stress, however whether long‐term changes in pro‐ or anti‐oxidant enzymes persist after cessation of treatment is not known. The purpose of this study was to examine the expression of pro‐ and anti‐oxidative enzymes to further examine their role in the cardiac protection conferred by transient ACE inhibition. Adult male spontaneously hypertensive rats (SHR) were divided into two groups with the control group (CL) receiving vehicle and the experimental group (EL) receiving enalapril (30 mg/kg per day) for two weeks, followed by a 2‐week washout. All rats were then treated with L‐NAME for 0 or 10 days. Pro‐oxidant enzyme NOX‐2 and anti‐oxidant enzymes SOD‐1, SOD‐2 and catalase levels were determined in left ventricle via Immuno‐Blotting. Two weeks after stopping enalapril, NOX‐2 levels tended to be increased over control. In response to L‐NAME, there was a significant increase in NOX‐2 in control (CL: 115%, p<0.05), but not EL (−4%, p>0.05) hearts over baseline control levels. Catalase, SOD‐1, and SOD‐2 levels were not persistently altered by transient ACE inhibition, nor were they affected by L‐NAME in either group. Taken together, the previously‐described long‐term protection afforded by transient ACE inhibition is not due to sustained up‐regulation of anti‐oxidant enzymes. Future studies will determine whether the lack of increase in NOX‐2 levels in the hearts of EL rats is a cause or consequence of the reduced fibrosis and inflammation.

Support or Funding Information

Springboard Grant, UA College of Medicine – Phoenix

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ganglioside GQ1b induces dopamine release through the activation of Pyk2
Authors: Chen Nai‐hong , Chu Shi‐feng , Zhang Zhao , Wang Zhen‐zhen
Published: June 2021

Growing evidence indicates that GQ1b, one of the gangliosides members, contributes to synaptic transmission and synapse formation. Previous studies have shown that GQ1b could enhance depolarization induced neurotransmitter release, while the role of GQ1b in asynchronous release is still largely unknown. Here in our result, we found low concentration of GQ1b, but not GT1b or GD1b (which were generated from GQ1b by plasma membrane‐associated sialidases), evoked asynchronous dopamine (DA) release from both clonal rat pheochromocytoma PC12 cells and rat striatal slices significantly. The release peaked at 2 min after GQ1b exposure, and lasted for more than 6 min. This effect was caused by the enhancement of intracellular Ca(2+) and the activation of Pyk2. Inhibition of Pyk2 by PF‐431396 (a dual inhibitor of Pyk2 and FAK) or Pyk2 siRNA abolished DA release induced by GQ1b. Moreover, Pyk2 Y402, but not other tyrosine site, was phosphorylated at the peaking time. The mutant of Pyk2 Y402 (Pyk2‐Y402F) was built to confirm the essential role of Y402 activation. Further studies revealed that activated Pyk2 stimulated ERK1/2 and p‐38, while only the ERK1/2 activation was indispensable for GQ1b induced DA release, which interacted with Synapsin I directly and led to its phosphorylation, then depolymerization of F‐actin, thus contributed to DA release. In conclusion, low concentration of GQ1b is able to enhance asynchronous DA release through Pyk2/ERK/Synapsin I/actin pathway. Our findings provide new insights into the role of GQ1b in neuronal communication, and implicate the potential application of GQ1b in neurological disorders.

Support or Funding Information

This work was supported by the National Natural Science Foundation of China (81730096, U1402221), CAMS Innovation Fund for Medical Sciences (CIFMS) ( 2016‐I2M‐1‐004), the Scientific Research Foundation of the Higher Education Institutions of Hunan Province (15K091), Beijing Key Laboratory of New Drug Mechanisms and Pharmacological Evaluation Study (BZ0150), PUMC Graduate Education and Teaching Reform Project (10023201600801).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Traditional Herbal Formula, CA01, in Mice suppresses Lung Inflammation induced by Cigarette Smoke
Authors: Park Eunsook , Jeon Woo‐Young , Seo Chang‐Seob , Ha Hyekyung , Jin Seong Eun , Kim Jinhee
Published: June 2021

Chronic obstructive pulmonary disease (COPD) is a progressive lung disease that is characterized by abnormal inflammatory responses. CA01 has been used widely as a traditional herbal medicine for the treatment of gastrointestinal symptoms in Korea, Japan and China. In this study, we examined the therapeutic effects of CA01 on the airway inflammatory response using mice model of cigarette smoke exposure together with lipopolysaccharide (LPS) treatment. During 4 weeks, mice were exposed to cigarette smoke for 1 h on 5 days per week and instilled intranasally with LPS after smoke exposure twice. CA01 was administered by oral gavage to mice on the same 5 days. In bronchoalveolar lavage fluid (BALF), CA01 administered groups significantly decreased the numbers of inflammatory cells and production of cytokines compared with cigarette smoke combined with LPS exposed groups. Furthermore, CA01 administration reduced histologically the infiltration of inflammatory cells into lung tissues and lung fibrosis. Our data suggest that CA01 exhibits anti‐inflammatory activity against cigarette smoke combined with LPS‐induced lung inflammation. Therefore, CA01 might be a potential drug for the treatment of lung inflammatory disease such as COPD.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Phoenix dactylifera leaf extract protects from paracitamol‐induced oxidative stress in rats
Authors: Salem Gamal A , Shaban Ahmed , Diab Hussain , Elsaghayer Wesam , Mjedib Manal , Hnesh Aomassad , Sahu Ravi P
Published: June 2021

Objectives

To determine the effects of leaf extract of Phoenix dactylifera against the oxidative stress response induced by a wide‐spectrum analgesic paracetamol in rats.

Methods

Adult Sprague dawley rats were treated with or without paracetamol (1500mg/kg) and leaf extract of Phoenix dactylifera (300mg/kg). Serum samples were collected to evaluate liver function tests, and hepatic tissue samples were obtained to assess oxidative stress‐related markers and histopathological examination.

Results

Phoenix dactylifera ameliorated paracetamol‐induced elevated serum liver markers such as alkaline phosphatase (ALP), aminotransferases (AST), alanine aminotransferase (ALT) and bilirubin. In addition, Phoenix dactylifera restored paracetamol‐induced reduced levels of liver tissue antioxidant enzymes such as glutathione peroxidase (GPx), catalase (CAT), and superoxide dismutase (SOD). Histological analysis demonstrate that Phoenix dactylifera conserved the hepatic tissue against lesions such as inflammation, centrilobular necrosis, and hemorrhages induced by PCM.

Conclusion

These findings indicate that Phoenix dactylifera possesses protective effects against paracetamol‐induced liver injury and oxidative stress, and could be used to explore against drug induced deleterious effects in experimental models.

Support or Funding Information

No funding support

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Experimentally‐Driven Molecular Docking via a Hybrid Approach
Authors: Roberts Arthur
Published: June 2021

High Ambiguity‐Driven biomolecular DOCKing (HADDOCK) protocols with CNS/Xplor and AutoDock Vina are powerful tools for drug receptor docking. AutoDock Vina performs blind drug docking and virtual drug screening very efficiently. Unfortunately, in many cases, the bound drug conformations determined by AutoDock Vina does not necessarily correlate well to experiments. On the other hand, docked conformations by HADDOCK are driven by experimental data combined with molecular dynamics, so they tend to correlate more to the experimental data. Unfortunately, this approach is considerably less efficient than AutoDock Vina and small molecules can become trapped in local energy wells during initial stages of docking. As a result, a seamless Python wrapper called HADDOCK‐Vina combining AutoDock Vina, CNS/Xplor, HADDOCK, MSMS, OpenBabel, ProPKA, Reduce and Volume Assessor together in a single package was developed. The experimental data is inputted in a simplified format with a few lines in HADDOCK‐Vina and produces a distance restraint file to be used in CNS/Xplor that has thousands of lines. The receptor is automatically processed and protonated by ProPKA within the wrapper. The HADDOCK‐Vina automatically produces and AutoDock search box over the entire receptor and a customized version AutoDockVina completely covers the surface of the receptor with ligand molecules. These bound ligands are filtered using the experimentally‐derived distance restraints and the experimentally‐related scoring algorithm of HADDOCK called the HADDOCK score (Hscore). By default, the three bound ligands with the lowest Hscore and the lowest energy are automatically refined further by energy minimization and simulated annealing using the HADDOCK protocols within CNS/Xplor. In this work, to demonstrate the effectiveness of the software, we show that we can reproduce ligand‐bound X‐ray protein crystal structures using only experimentally‐derived distance restraints. We also compare the wrapper to the individual software programs.

Support or Funding Information

This work is supported by NIGMS NIH R15 (R15GM107913‐01A1) and NCI NIH R01 (R01CA204846‐01A1) grants.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Confounding effect of UGT1A1 interindividual variability on inhibition of its in vitro activity by kinase inhibitors.
Authors: Alade Nathan , Basit Abdul , Zhang Haeyoung , Bhatt Deepak , Collins Carol , Prasad Bhagwat
Published: June 2021

Background and Objective

Inhibition of UGT1A1 by kinase inhibitors (KIs) has been shown to cause mild to serve hyperbilirubinemia in cancer patients1. UGT1A1 is highly polymorphic and displays variable expression. The specific aims of this study were i) to stratify human liver microsomes (HLMs) based on UGT1A1 abundance, activity and genotype, ii) to screen 10 clinically used KIs for their UGT1A1 inhibition potential, and iii) to determine inhibition potential (IC50) of the top three potent KIs in high and low UGT1A1 expressing HLM samples.

Methods

Targeted genotyping of liver tissue donors (N=445) was performed; and UGT1A1 protein abundance and activity in HLM samples were quantified using a validated proteomics method and using bilirubin or estradiol as probe substrates, respectively. The mono and di‐bilirubin glucuronides and estradiol 3‐β‐D‐glucuronide (E3βG) were monitored by LC‐MS/MS. Association analysis of UGT1A1 genotype with protein abundance or activity was performed. Screening of UGT1A1 inhibition by KIs (at 1 and 20 uM) was performed using a high UGT1A1 expressing (46.7 ± 4.6 pmol/mg) HLM sample. IC50 values of KIs for UGT1A1 were determined using above mentioned high and a low (10.1 ± 1.6 pmol/mg) expressing HLM samples.

Results

UGT1A1 abundance and activity exhibited age‐dependent maturation with 8‐fold increase from neonates to adults. UGT1A1 SNPs rs10929302, rs887829, rs111741722 and rs4124874, are associated with decreased UGT1A1 abundance and activity, whereas rs8175347 was only associated with decreased abundance. Glucuronidation of both substrates was inhibited by the KIs in a substrate‐independent manner in the following order (% inhibition of E3βG activity at 1 and 20 μM in parenthesis): dasatanib (46 and 91)> sorafenib (11 and 74)> temsirolimus (50 and 71)> sirolimus (0 and 63) > sunitinib (21 and 38) > nilotinib (22 and 34) > gefitinib (0 and 29) > pazopanib = imatinib = lapatinib (0). The top three KIs showed differential IC50 values for E3βG inhibition in high and low UGT1A1 expressers, i.e., dasatanib (2.0, and 4.0), sirolimus (IC50= 7.6, and 32.6), and sorafenib (0.95, and 18.0), respectively.

Discussion

UGT1A1 abundance is highly variable with significant effect of age and genotype. Inter‐individual variability in UGT1A1 plays a confounding role in predicting drug‐endobiotic or drug‐drug interactions (DDIs) mediated by this enzyme. Identification of genetic or non‐genetic factors associated with UGT1A1 activity will allow better prediction of UGT1A1 mediated drug metabolism and toxicity.

Support or Funding Information

This work was primarily supported by the National Institutes of Health [Grant no. R01 HD081299].

NA and AB contributed equally to this work.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Inhibition of CYP3A7 by Ritonavir and Lopinavir Leads to Adrenal Insufficiency in the Developing Infant by Blocking DHEA‐S Oxidation
Authors: Kandel Sylvie E. , Han Lyrialle W. , Mao Quingcheng , Lampe Jed N.
Published: June 2021

CYP3A7 is the predominant CYP3A isoform in the neonate and developing infant. Its primary function is the 16α‐hydroxylation of the endogenous hormone DHEA‐S, which is important for downstream maintenance of hormone homeostasis in the adrenal gland. Newborns infected with HIV are typically treated with a combination therapy of Lopinavir and the pharmacokinetic enhancer Ritonavir. However, adrenal insufficiency due to increased DHEA‐S concentrations, and premature birth, have been reported in this treatment population, leading to adverse outcomes in this fragile patient group. Given that Ritonavir is a known potent inhibitor of CYP3A4, we hypothesized that it might also be an inhibitor of CYP3A7‐mediated DHEA‐S oxidation and that this inhibition might be responsible for the adrenal insufficiency and other side effects associated with Lopinavir‐Ritonavir treatment. Indeed, we observed significant inhibition of CYP3A7 DHEA‐S hydroxylation activity in the presence of Ritonavir with an IC50 of 0.05 μM. Furthermore, Ritonavir displayed time‐dependent inhibition of DHEA‐S oxidation, with a Ki of 0.336 μM and a Kinact of 0.1077 min−1, suggesting that it may be a mechanism based inactivator of CYP3A7. Lopinavir was also capable of inhibiting CYP3A7, albeit to a much lesser extent than Ritonavir with an IC50 of 5.88 μM for DHEA‐S 16α‐hydroxylation, and without the associated time‐dependency. Interestingly, Lopinavir alone has a much higher intrinsic clearance rate in fetal liver microsomes compared to adult liver microsomes, indicating the need for a pharmacokinetic enhancer adjuvant in order to slow clearance and maintain efficacy. However, our results suggest that closer monitoring of the effective dose of the pharmacokinetic enhancer may be required in order to prevent significant inhibition of CYP3A7 DHEA‐S oxidation and the associated adverse effects.

Support or Funding Information

This work was supported by the National Institutes of Health, Institute of General Medical Sciences [Grant P20‐GM103549‐07]/ National Center for Research Resources [Grant P20‐RR021940] and Kansas IDeA Network of Biomedical Research Excellence [Grant QH846868‐KINBRE] to JNL.

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