bluuebunny/crossref_metadata_2025 huggingface dataset:
Exposures to L‐felinine Suppressed Plasma Testosterone in Laboratory Rats
Authors: Voznessenskaya Vera V , Laktionova Tatiana K
Published: June 2021

Felinine is a unique sulfur‐containing amino acid found in the urine of domestic cats and select members of the Felidae family (Rutherfurd et al. 2002). L‐felinine and its sulfur‐containing volatile derivatives excreted with urine in sex‐ and age‐dependent manner (Miyazaki et al. 2006). These cat‐specific volatile compounds may represent pheromones used as territorial markers for conspecific recognition or reproductive purposes by mature cats (Miyazaki et al. 2008). Species‐specific compounds may be used also by other species to recognize potential predators and their physiological status. In our previous studies we showed that exposures of rats from laboratory heterogenous population during gestation to the cat urine or L‐felinine significantly affected reproductive output. We observed significantly lower weight of newborn pups and skewed sex ratio in favour in males. At the behavioral level, exposed to cat urine/L‐felinine male rats showed fewer mountings and mountings with intromissions in presence of receptive females. In the current study we examined the influence of domestic cat‐specific compound L‐felinine on testosterone secretion in male rats. Intact cat urine was used as positive control; tap water was used as negative control. Test subjects were 3–4 month old Norway rats Rattus norvegicus from laboratory population. Males were placed singly two weeks prior to experiment. We used concentration of L‐felinine comparable to naturally occurring in the adult cat urine (Rutherfurd et al. 2002). Cotton balls soaked with L‐Felinine (0.05% w/v, 0.2 ml, US Biologicals), cat urine (0.2 ml) or tap water (0.2 ml) in plastic perforated containers were placed directly into home cages of rats. We renewed test solutions twice a week. Experimental animals were randomly assigned to one of treatment groups. Blood samples (50 μl) were obtained from rear foot pad (Miller et al. 1997). Plasma testosterone was assayed (in duplicate) by enzyme immunoassay method (EIA‐1559, DRG). Exposure of male rats to cat urine (0.2 ml) during two weeks significantly suppressed plasma testosterone (p=0. 0020, n=10, t‐test). Analogous exposures to L‐felinine (0.05 %, 0.2ml) produced quite similar effect (p =0.049, n=10, t‐test) in male rats. The effect was dose dependent: we did not observe suppression of plasma testosterone when lower amount of L‐felinine solution (0.05 %, 0.1ml.) was applied to home cage of male rats. It appears to be that altered sexual behavior of male rats exposed to L‐felinine/cat urine could be explained by significantly lowered testosterone secretion. The expression of the secondary defense reactions by laboratory naive rats and the failure tohabituate at the hormonal level may indicate an innate nature of the response.

Support or Funding Information

Supported by Russian Foundation for Basic Research grant 19‐34‐70024 mol_a_mos

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Integrin receptor α4β7+ CD4 T cells promote mucosal and hepatic inflammation in NASH
Authors: Raeman Reben , Anania Frank A , Parkos Charles , Nusrat Asma , Kumar Pradeep , Desai Chirayu , Iyer Smita S , Liu Yunshan
Published: June 2021

There is mounting evidence that impairment of the intestinal epithelial barrier underlies the pathogenesis of non‐alcoholic steatohepatitis (NASH); but, the specific mechanisms by which this occurs are unknown. Using F11r−/− mice, a murine model which exhibits altered intestinal permeability, we investigated the role of gut‐homing integrin receptor α4β7 expressing CD4 T cells in diet‐induced loss of intestinal epithelial barrier. Fed a WD for eight weeks, F11r−/− mice develop features of severe NASH, the result of damage to the integrity of the intestinal epithelium induced by the WD. WD‐fedmice demonstrated a systemic increase in IFNγ and TNFaproducing α4β7+ CD4 T cells, with greater recruitment of α4β7+ CD4 T cells in the intestine of F11r−/− mice relative to controls. This correlated with a markedly higher expression of integrin receptor α4β7 ligand, mucosal addressin cell adhesion molecule 1 (MAdCAM‐1), in the colonic mucosa of F11r−/− mice fed the WD. Elevated colonic expression of MAdCAM‐1 correlated with increased mucosa‐associated Proteobacteria in F11r−/− mice, but not in control mice. Depletion of microbiota reduced colonic MAdCAM‐1 expression and attenuated mucosal inflammation suggesting microbiota promote MAdCAM‐1 expression in the colon. Abolishing α4β7/MAdCAM‐1 interaction using an α4β7 monoclonal antibody significantly decreased α4β7+ CD4 T cell recruitment to the intestine and reduced colonic inflammation. This intervention also attenuated hepatic inflammation and fibrosis, and improved indices of the metabolic syndrome by reducing hepatic α4β7+ CD4 T cell recruitment driven by diet‐induced, microbiota‐dependent hepatic MAdCAM‐1 expression. Together, these data provide the first direct evidence that α4β7+CD4 T cells regulate intestinal barrier and hepatic inflammation in NASH.

Support or Funding Information

Research reported in this publication was supported by the National Institute of Diabetes and Digestive and Kidney Diseases of the National Institutes of Health under Award Number K01DK110264 to RR. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health, the US Food and Drug Administration, the US Department of Health and Human Services, or the US Government.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Impact of Single Nucleotide Polymorphisms on HPA‐Axis Functionality
Authors: Caruso Alexa P , Sharma Tanuj , Martino Audrey , Zhou Helen , Cohen Justin R. , Lukowitsky Mark , Cohen Brian D
Published: June 2021

The hypothalamic‐pituitary‐adrenal (HPA) axis is responsible for regulating the secretion of the glucocorticoid hormone cortisol in response to stress. Diseases of cortisol dysregulation, such as Cushing's syndrome (hypercortisolism) and Addison's disease (hypocortisolism), share a common symptom of depression. Based on this we, and others, have hypothesized that single nucleotide polymorphisms (SNPs) in the glucocorticoid receptor (GR), the mineralocorticoid receptor (MR), and other genes associated with cortisol regulation may contribute to depression. Our study investigated the genotypic frequency of several SNPs that affect GR and MR sensitivity to cortisol in a clinical population. DNA samples were acquired via buccal swab from patients suspected of suffering from depression. Extracted DNA was analyzed using a novel multiplex allele‐specific polymerase chain reaction assay to determine the allelic frequency of SNPs associated with hypersensitivity or resistance to cortisol. Saliva samples were also collected from these patients and analyzed by ELISA to measure cortisol levels. In addition, patients completed multiple measures of depression and anxiety.

Preliminary results showed that mutant haplotypes of three specific alleles in the GR, BclI (rs41423247), N363S (rs56149945), and TthlllI (rs10052957), were seen more frequently in the patient population compared to the general population. Although depression (as measured by the Beck Depression Inventory) and salivary cortisol did not change significantly between clinic visits, there was a significant decrease in state anxiety (as measured by the State‐Trait Anxiety Inventory) between visits for patients with the wild type or heterozygous BclI haplotypes while patients with the homozygous mutant haplotype did not show an improvement in state anxiety. This is the first demonstration that genotypic variation in cortisol sensitivity is associated with changes in state anxiety. This could lead to more specific and successful treatments for depression and anxiety.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Phrenic Nerve and Carotid Body Responses to Hypoxia and CO2 in Naked Mole Rats
Authors: Wang Benjamin , Peng Ying‐Jie , Nanduri Jayasri , Wang Ning , Prabhakar Nanduri R , Pamenter Matthew
Published: June 2021

Naked mole‐rats (NMRs), which live in underground burrows, are tolerant to hypoxia. In the present study, we examined the efferent phrenic nerve and carotid body (CB) responses to hypoxia and CO2 in NMRs and the results were compared with Bl6 mice. The phrenic nerve responses to 10% O2 and 10% CO2 for 3 min were examined in anesthetized animals. Stimulation of minute neural respiration (MNR = phrenic bursts/min × tidal phrenic amplitude) evoked by hypoxia was ~ 50% less, whereas by CO2 was ~200% higher in NMRs as compared to Bl6 mice. The magnitude of CB sensory nerve excitation for a given level of hypoxia was comparable, whereas the CB response to CO2 was ~100% higher in NMRs as compared with Bl6 mice. Morphological analysis of CB with anti‐chromogranin A (CGA, a synaptic vesicle marker) and anti‐tyrosine hydroxylase (TH; a glomus cell marker) antibodies showed a dispersed pattern of glomus cells in NMR CBs as compared with a clustered pattern in Bl6 CBs. These findings suggest that breathing and CB responses to hypoxia and CO2 differ between NMR and Bl6 mice and these effects are associated with striking differences in CB morphology.

Support or Funding Information

Supported by NIH‐HLBI P01‐HL‐90554

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
H2S synthesis inhibitor prevents hypoxia‐evoked periodic breathing in spontaneous hypertensive rats
Authors: Zhang Xiuli , Peng Ying‐Jie , Nanduri Jayasri , Muzzio Miguel , McCormick David L , Prabhakar Nanduri R
Published: June 2021

Hyperactive carotid body (CB)‐chemo reflex has been implicated in evoking periodic breathing (PB) characterized by episodes of hyperventilation and hypoventilation (decreased tidal volume and respiratory rate). Spontaneous hypertensive (SH) rats exhibit augmented CB response to hypoxia and this effect is mediated by increased H2S production from cystathionine‐g‐lyase (CSE; Peng et al. PNAS, 2017). In the present study, we examined whether acute hypoxia evokes PB in SH rats. Breathing was monitored in conscious SH rats (age 2–4 months old) by whole‐body plethysmography. Hypoxia evoked PB in SH rats in a stimulus‐dependent manner. Exposure to 12% O2 elicited 40 ± 3 episodes of PB/hr and severe hypoxia (8–10% O2) elicited 114 ± 4 episodes of PB/hr. Oral administration of L‐propargylglycine (L‐PAG), a CSE inhibitor, prevented CB hypersensitivity to hypoxia and stabilized breathing in dose‐related fashion. L‐PAG is rapidly absorbed after oral dosing (Tmax = 0.25 to 0.5 hr) and oral bioavailability is 100%. These data demonstrate that CSE inhibition provides a viable mechanism for prevention of breathing instability arising from CB hypersensitivity to hypoxia.

Support or Funding Information

Supported by NHLBI grant UH3HL‐123610

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Induction of asthma causes sensitization of the carotid bodies to lysophosphatidic acid
Authors: Jendzjowsky Nicholas Gregory , Roy Arijit , Wilson Richard
Published: June 2021

Previously, we demonstrated an important role for the carotid bodies in asthmatic bronchoconstriction. Specifically, we showed that an allergen‐induced increase in circulating lysophosphatidic acid (LPA) in asthmatic rats activates LPA receptors (LPAr) in the carotid bodies and post‐ganglionic TRPV1 channels, provoking vagal activity and thereby triggering acute bronchoconstriction (Jendzjowsky et al. 2018 Nat Comm. 9, 4030). Here we report that induction of asthma causes molecular and functional changes to the carotid bodies that sensitize them to LPA.

Experiments were carried out using the standard ovalbumin‐sensitized brown Norway rat model of asthma. Carotid bodies and petrosal ganglia were extracted 3 hours after the third ovalbumin challenge in asthmatic (OVA) or third saline challenge in naïve (N) rats. Using qPCR (Δctt OVA vs N), LPAr expression was increased in the carotid body (LPAr1: 556x; LPAr2: 20x; LPAr3: 2x; LPAr4: 150x; LPAr5 only detected in OVA: LPAr6: 175x; TRPV1 not detected, as demonstrated previously Roy et al. 2012 J Appl Physiol 112:212–224) whereas LPAr 1–6 and TRPV1 were similarly expressed in petrosal ganglia of OVA and N rats. Using the perfused ex vivo carotid body preparation, baseline normalized carotid sinus nerve responses to increasing doses of LPA activity, were significantly augmented in OVA compared to N rats (2.5uM‐ OVA: 1.12 ± 0.01 N: 1.05 ± 0.02, p=0.162; 5uM: OVA: 1.23 ± 0.03 N:1.10 ± 0.02, p=0.009; 10uM: OVA: 1.40 ± 0.07 N: 1.17 ± 0.03, p<0.001). In contrast, carotid body activity in response to hypoxia was not different between OVA (2.54 ± 0.47) and N (1.95 ± 0.10) rats (p=0.243).

Our data suggest that asthmatic carotid bodies have enhanced responsiveness to LPA, independent of hypoxic sensitivity. These changes likely contribute to the acute allergen‐induced bronchoconstriction which is the hallmark of asthma that appears to be mediated, in part, by a novel carotid body‐vagal‐lung neuronal pathway. Targeting carotid body LPA signalling may reduce asthmatic bronchoconstriction and provide new pharmaceutical targets for inflammatory lung disease.

Support or Funding Information

This project was funded by The Lung Association and Canadian Institute for Health Research.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Ventilatory response to hypoxia and tolerance to high altitude in women: Influence of menstrual cycle, oral contraception and menopause
Authors: Richalet Jean‐Paul , Lhuissier François J , Jean Dominique
Published: June 2021

Tolerance to high altitude in women might be influenced by hormonal status since female hormones are known to modulate ventilation. We explored the influence of 1) the phase of menstrual cycle, 2) oral contraception, 3) menopause with or without hormonal treatment, on ventilatory (HVRe) and cardiac (HCRe) responses to hypoxia at exercise in 1060 women prior to exposure at high altitude. HCRe and HVRe were measured during a routine hypoxia exercise test. Prevalence of Severe High Altitude Illness (SHAI) was assessed in a subgroup of 260 women. Four groups of women were defined: non‐menopaused with or without oral contraception; menopaused with or without hormonal treatment. In non‐menopaused women without contraception, HVRe was higher in the early luteal phase than in the early follicular phase (0.89±0.37 vs 0.75±0.27 ml/min/kg, p=0.03). HVRe was higher (p=0.016) and HCRe lower (p<0.001) in menopaused vs non‐menopaused women. Independently of hormonal status, HVRe decreased from second to fourth decade of age and increased from fourth to eighthdecade, while HCRe consistently decreased with ageing. Oral contraception or hormonal treatment had no effect on responses to hypoxia. The prevalence of SHAI was similar in all groups. SHAI prediction score was higher and HVRe lower in women with SHAI, without taking actetazolamide. In conclusion, 1) physiological responses to hypoxia depends on the ovarian cycle phase and on menopause status, 2) oral contraception, and hormonal treatment have no influence on response to hypoxia and tolerance to high altitude, 3) independently of hormonal status, ageing modulates physiological responses to hypoxia, 4) the optimal phase for high altitude acclimatization for women would be the early luteal phase.

Support or Funding Information

Association pour la Recherche en Physiologie de l'Environnement

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Intermittent Hypoxia, Sleep Fragmentation and Glucose Tolerance
Authors: Framnes‐DeBoer Sarah , Arble Deanna
Published: June 2021

Obstructive sleep apnea is a common sleep disorder affecting 17% of the total population and characterized by repeated bouts of intermittent hypoxia and sleep fragmentation. Both intermittent hypoxia and sleep fragmentation are hypothesized to have independent consequences on cardiometabolic health. In the clinical setting, the ability to tease apart the relative contributions of intermittent hypoxia from sleep fragmentation is limited. Here, we use mice exposed to intermittent hypoxia either during their rest or active phase to systematically determine the metabolic effects of intermittent hypoxia + sleep fragmentation or intermittent hypoxia alone. We find that 9 hours of mild, intermittent hypoxia, (a 30‐second, 12% 02 event occurring once per minute) is sufficient to impact glucose metabolism when applied during the resting phase, but not during the active phase (AUC 16733 ±1326 vs 21189 ±1395, respectively, p < 0.05). However, only a more severe intermittent hypoxia protocol (a 30‐second, 7% 02 event occurring once per minute) was found to affect the glucose tolerance of both resting and awake mice (AUC: 17063 ± 1328 vs 17167 ±696, respectively). These data demonstrate that the severity of intermittent hypoxia dictates the glucose response in awake mice, while sleep fragmentation may chiefly influence the glucose metabolism of a resting mouse. Taken together, these results demonstrate distinctive and interdependent roles for intermittent hypoxia and sleep fragmentation on glucose metabolism.

Support or Funding Information

Dr. Arble receives funding from the American Heart Association (PI Arble, 17SDG33660108) and Marquette Biological Sciences Department.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Hypoxia Inducible Factor Activation Prevents Renal Mitochondria Dysfunction and Improves Cortical Oxygenation in Type 1 Diabetic Mice
Authors: de Carvalho Carla Luisa Lopes , Schiffer Tomas , Zheng Xiao‐Wei , Grunler Jacob , Karlsson Susanne , Mazzone Massimiliano , Carmeliet Peter , Inagi Reiko , Hansell Peter , Catrina Sergiu‐Bogdan , Friederich‐Persson Malou , Palm Fredrik
Published: June 2021

Hypoxia‐inducible factors (HIFs) are transcription factors that are activated during hypoxia in order to regulate a hypoxic gene response. During normoxia, prolyl hydroxylase domain‐containing protein 2 (PHD2) is one of three isoforms that acts on HIF as a first step towards its degradation and its knockdown consequently results in HIF activation. Previous studies have demonstrated that diabetes induces mitochondria dysfunction resulting in renal cortical hypoxia, yet studies indicate that hyperglycemia impairs HIF stabilization resulting in suboptimal activation and an inability to correct renal hypoxia. In this study, we hypothesize that increased HIF‐activation resulting from PHD2 knockdown, prevents mitochondria dysfunction and improves renal oxygenation in type 1 diabetic mice.

Balb/c mice from four groups (Wild type normoglycemic (n=16), wild type diabetic (n=10), PHD2+/− normoglycemic (n=16) and PHD2+/− diabetic (n=11)) were placed in metabolic cages for 24 h urine collection followed by conscious determination glomerular filtration rate (GFR), using a single bolus injection of FITC‐inulin. Diabetes was induced by a low dose injection of strepzotocin (i.p.) for 5 consecutive days and diabetes duration was 5–6 weeks. Renal cortical oxygenation was measured with Clark electrodes and kidney mitochondria function, including mitochondria proton leak, was assessed by high resolution respirometry. Urinary excretion of albumin and thiobarbituric acid reactive species were measured and corrected for 24 h urine volume.

Diabetes resulted in increased GFR in both groups compared to normoglycemic mice. Wild type diabetic mice presented with increased total mitochondrial leak respiration compared to normoglycemic mice, which was prevented in the PHD2+/− mice. Interestingly, regulated leak, defined as mitochondrial proton leak regulated by uncoupling proteins and adenosine nucleotide translocase, was increased in diabetic PHD2+/− only, while unregulated leak, defined as total leak minus regulated leak, was increased in wild type diabetic mice. Both diabetic groups displayed increased albuminuria and thiobarbituric acid reactive species compared to normoglycemic control mice.

In conclusion, activation of HIF by reducing PHD2 expression prevents mitochondria dysfunction and renal cortical hypoxia in type 1 diabetic mice. This could imply a novel target for the PHD inhibitors currently in clinical trials.

Support or Funding Information

This study was supported financially by the Swedish Research Council, the Swedish Diabetes Foundation and the Family Ernfors Fund.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Glucosamine chronic treatment differently modulates uterine artery from pregnant and virgin Wistar rats
Authors: Justina Vanessa Dela , Webb R Clinton , Lima Victor Vitorino , Giachini Fernanda Regina
Published: June 2021

Background

O‐linked β‐N‐acetylglucosamine (O‐GlcNAc) is a post‐translational modification that modulates several proteins related to the vascular tone control. Dysregulation of O‐GlcNAcylation occurs in a wide range of diseases, favoring endothelial dysfunction. During pregnancy, vascular dysfunction observed in the uterine vasculature results in inadequate nutritional support between the maternal‐fetal interface. It remains to be elucidated whether augmented O‐GlcNAc levels impact uterine artery (UA) function from virgin or pregnant rats.

Hypothesis

Increased O‐GlcNAc impairs the endothelial function of the uterine artery.

Methods

Pregnant or virgin Wistar rats (14‐week old) were treated either with glucosamine (300 mg/Kg) or vehicle, for 21 days, i.p. The UAs were rapidly removed, cleaned from fat tissue and mounted for isometric force development record. Concentration response‐curves to acetylcholine (1 nM to 100 μM) were performed in UA contracted with phenylephrine (10 μM), in the presence or absence of L‐NAME (100 μM, 30 minutes). Protein expression was evaluated by western blot.

Results

Glucosamine treatment increased O‐GlcNAc expression in UAs from virgin rats (p= 0.04). Decreased endothelium‐dependent relaxation was observed in UAs from treated virgin rats compared to vehicle [Emax 56.16 ± 4.18 vs. 76.38 ± 3.92%, respectively], which was abolished by incubation with L‐NAME. Endothelial nitric oxide synthase (eNOS) activity was decreased (p=0.04) by glucosamine‐treatment in virgin rats. On the other hand, in pregnancy, treatment with glucosamine decreased O‐GlcNAc expression (p=0.03) in UAs compared to vehicle. Increased sensitivity to acetylcholine [pD2 6.43 ± 0.13 vs. 6.90 ± 0.11] was observed in the UAs of glucosamine‐treated pregnant rats compared to vehicle pregnant rats with no differences in the maximal response. eNOS activity was unchanged between these groups.

Conclusions

Glucosamine‐treatment impairs UA endothelial function in virgin, but not in pregnant rats.

Support or Funding Information

CAPES

FAPEG

NIH (HL‐134604)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
In Vivo Effect of Luteolin during Oxaliplatin Treatment for Colorectal Cancer
Authors: Moon Nayoung , Jang Chan Ho , Averilla Janice Nullan , Jeong Soojung , Lim Ji Sun , Lo Jisun , Kim Jong‐Sang
Published: June 2021

Phytochemicals that induce nuclear factor (erythroid‐derived 2)‐like 2 (Nrf2) can lower cellular oxidative stress and thus favor cell viability. Considering our previous in vitro study demonstrating that strong antioxidative called luteolin interfered with the anticancer effect of oxaliplatin on HCT116, we hypothesized that luteolin administration would be unadvantageous during oxaliplatin‐based chemotherapy. To test this hypothesis, HCT116 xenograft mouse model was established in BALB/c nude mice. Once palpable tumors were developed after HCT116 cells were subcutaneously transplanted on both flanks of each mouse, mice were randomly allocated into four groups that were intraperitoneally injected with (1) vehicle only (control), (2) oxaliplatin at a dose of 10 mg/kg body weight (BW), (3) luteolin at 50 mg/kg BW, or (4) combination of luteolin and oxaliplatin. Tumor size and body weight were regularly monitored (three times a week) for three weeks. Results demonstrated that oxaliplatin treatment inhibited the growth of xenografted tumors as expected, luteolin treatment had little effect on the tumor size, and combinatorial treatment of oxaliplatin and luteolin most suppressed tumor growth. In addition, oxaliplatin‐luteolin combo reduced heme oxygenase‐1 (HO‐1) protein expression compared to oxaliplatin alone. Moreover, luteolin alone treatment enhanced protein expressions of p53 and its downstream molecule p21. These findings were suggestive of in vivo synergistic effect of combinatorial treatment of oxaliplatin and luteolin at the tested doses in this study and the possible coordinated regulation via p53 and/or HO‐1‐mediated cytoprotection in HCT116 xenograft tumor growth. According to the current observations, our hypothesis ought to be reconsidered; luteolin could advantageously hamper tumor growth during oxaliplatin treatment in colon cancer.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Previous Adaptation Period Modulates Pregnancy Swimming Neuroprotective Effects on the Offspring Following Neonatal Hypoxic‐Ischemic Brain Injury
Authors: Tassinari Isadora Davila , Sanches Eduardo Farias , Fabres Rafael Bandeira , Andrade Mirella Kielek Galvan , Rodrigues Thais Lopes , Carvalho Andrey Soares , Rezena Ester , Mazeron Larissa Sagaz , Netto Carlos Alexandre , de Fraga Luciano Stürmer
Published: June 2021

Neonatal hypoxic‐ischemic encephalopathy (HI) is one the main causes of neurological damage in newborns. Pregnancy has emerging as a feasible therapeutic window for preventing central nervous system damage and has been shown to decrease early and long‐term HI consequences. However, experimental literature is conflicting and point that variables, such as a previous adaptation period to the exercise (before mating) has decisive impact on swimming consequences to the injured offspring's brain.

Briefly, the adaptation period consisted of exposing female rats to 7 days (with increasing time exposure ‐ from 5 to 20 minutes) to a tank (200 cm diameter) filled with water at 32°C. After mating, from gestational day 0 to 21, pregnant rats were submitted to 20min/day of swimming or remained in the standard cages. After birth, seven‐days‐old rats were subjected to the Levine‐Rice HI model (right common carotid artery occlusion followed by 8% O2/60min). Western blotting was used to assess protein expression of GFAP (astrocytic reactivity), cytochrome c and cleaved caspase 3 (apoptotic markers) was performed in hippocampus 24h after HI. Animals were divided into 8 experimental groups, according adaptation, swimming and. Behavioral analysis was performed from PND45 and included sensorimotor and cognitive testing. Histological analysis assessed structural brain damage volumes of hemisphere and hippocampus. This study was approved by the local Institutional Animal Care and Use Committee (UFRGS #33497)

Non‐parametric tests showed significant increase in astrogliosis in non‐adapted HI animals only. Swimming decreased apoptotic cell death despite adaptation period. Cylinder test evidenced HI effects (increasing the use of the ipsilateral forepaw in HI animals). No effects of swimming nor adaptation were observed in the test. In the open field testing, only HI animals whose mothers where adapted previously to the injury had increased locomotion and swimming reversed the HI effects on the test. Hemisphere and hippocampus were preserved only in the HI group whose mothers swam and were adapted before mating. In summary, the adaptation period has a major role on pregnancy swimming effects following HI and must be considered in future studies and the mechanisms involved in such modulation need to be further explored.

Support or Funding Information

This study was supported by CNPq, FAPERGS and CAPES ‐ Brazil.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Notch1 Overexpression in Cardiac Mesenchymal Stem Cells Renders their Exosomes Highly Effective in Promoting Angiogenesis and Cardiac Regeneration
Authors: Xuan Wanling , Tang Yaoliang , Ashraf Muhammad
Published: June 2021

Objective

Notch1 signaling is critical in postnatal angiogenesis. Cardiac mesenchymal stem cells (C‐MSC) are endogenous cardiac stromal cells which are believed to facilitate cardiac regeneration via paracrine effects. Here, we investigated the effects of exosomes from Notch1 engineered C‐MSC on angiogenesis and cardiac myocyte proliferation in ischemic myocardium.

Methods and results

C‐MSC were isolated from conditional Notch1 knock out mice. For Notch1 knockout C‐MSC generation, Notch1 gene was deleted via adenoviral vector mediated Cre recombination. For Notch1 overexpression, cells were modified with Notch1 intracellular domain (NICD). Myocardial infarction (MI) mouse model was generated by LAD ligation. Exosomes from C‐MSC (Exo‐C‐MSC), Notch1 knockout C‐MSC (Exo‐C‐MSCNICD KO) and Notch1 overexpressing C‐MSC (Exo‐C‐MSCNICD) were isolated and characterized. Both Exo‐C‐MSC and Exo‐C‐MSCNICD were strongly pro‐angiogenic under in vitro and in vivo conditions. Compared with Exo‐C‐MSC, Exo‐C‐MSCNICD treatment led to dense tube formation and higher vessel density in peri‐infarct area. Moreover, knockout of Notch1 significantly abrogated the pro‐angiogenetic effect of Exo‐C‐MSC. Compared with control group, pretreatment of Exo‐C‐MSC and Exo‐C‐MSCNICD also exerted anti‐apoptotic effects on endothelial cells stressed with 300 μM H2O2, for 8h. Exo‐C‐MSCNICD KO had no anti‐apoptotic effect in H202 treated cells. As expected, intramyocardial injection of Exo‐C‐MSCNICD were strongly cardioprotective on cardiomyocytes (CMs) and vessel cells 24h after MI. Though both Exo‐C‐MSC and Exo‐C‐MSCNICD treatment improved cardiac function and decreased fibrosis in mice one‐month post‐MI, Exo‐C‐MSCNICD were more effective in improving cardiac function and reducing fibrosis. On the other hand, Exo from Notch1 knockout MSC had no effects on fibrosis or function. A significant effect on vessel density and CMs proliferation (ki67 and α‐actinin double positive cells) in peri infarct area was observed with the treatment of Exo‐C‐MSCNICD in comparison with PBS, Exo‐C‐MSC and Exo‐C‐MSCNICD KO groups.

Conclusion

Notch1 overexpression in C‐MSC produced highly effective exosomes in preventing cell death, promoting angiogenesis, cardiomyocyte proliferation and in restoring cardiac function after MI. These data suggest that Notch1 activation may further enhance the effectiveness of exosomes from C‐MSC in cell‐free therapy.

Support or Funding Information

R01 HL134354 and R01 AR070029

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
3D mitochondrial network formation during postnatal skeletal muscle development of mice
Authors: Kim Yuho , Lindberg Eric , Bleck Christopher KE , Glancy Brian
Published: June 2021

Mitochondrial connectivity plays an important role for muscle energy distribution. However, it is obscure how their structural formation is developed during postnatal muscle growth. Here, using a high‐resolution electron microscopy (i.e., FIB‐SEM), we have sought to examine three dimensional (3D) mitochondrial network formations in oxidative (soleus) and glycolytic (gastrocnemius) muscles of C57BL/6N mice at postnatal (P) 1, 7, 14, 21, and 42 day, respectively. After semi‐automated machine learning process, 3D mitochondrial reticulum formations were rendered and quantitatively analyzed by ImageJ software. Both types of skeletal muscles showed similarly fiber parallel mitochondrial network formations during early days (P1–P7). After P14, oxidative muscles developed grid‐like mitochondrial reticulum, while glycolytic muscle mitochondria were found as perpendicular formations along the Z‐lines. Unlikely to glycolytic muscles where individual mitochondrial volumes were similarly maintained across the development, oxidative muscles significantly increased those volumes by P14 and then largely decreased to the level comparable to glycolytic muscles. After P14, oxidative muscles maintained notably higher total mitochondrial volumes as compared to glycolytic muscles. Furthermore, we observed that at P14, glycolytic muscles showed remarkably higher ratio of mitochondrial surface area to volume, as well as sphericity, than oxidative muscles. Another interesting point was that during early muscle growth (P1–P7), mitochondrial elongation, aspect ratio, and length were all shown to be higher in glycolytic muscles than in oxidative muscles. In conclusion, this preliminary study provides how 3D mitochondrial networks are changed over the course of postnatal muscle development, and also suggests that P14 may be a critical time point for the mitochondrial transformation. However, further studies are needed to better understand 3D muscle mitochondrial development along with contractile components and other subcellular elements such as lipid droplets and sarcoplasmic reticulum.

Support or Funding Information

This work is supported by the Division of Intramural Research of the National Heart, Lung and Blood Institute and the Intramural Research Program of the National Institute of Arthritis and Musculoskeletal and Skin Diseases.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Astragaloside IV Increases Phosphorylation of GSK‐3β And S6K1 via The AKT Pathway in C2C12 Muscle Cells
Authors: Tuohy Kyle , Abboud Mohamad , Sitnick Mitchell
Published: June 2021

Astragaloside IV (AsIV) is the primary active component of the Astragalus membranaceus herb, used in traditional Chinese medicine to treat metabolic disorders. Through in vitro and in vivo experimentation, this saponin phytochemical has been shown to regulate glycogen synthesis through protein kinase B (AKT)‐mediated phosphorylation of glycogen synthase kinase 3 beta (GSK‐3β). Phosphorylation of GSK‐3β at Ser9 downregulates the enzyme's activity and promotes glucose uptake and responsiveness. Along with control of glucose homeostasis, AKT activation is an important regulator of protein synthesis via the downstream phosphorylation of mammalian target of rapamycin (mTOR) and ribosomal protein S6 kinase beta‐1 (S6K1). While AsIV has been shown to exhibit metabolic benefits, there is little data investigating the possible effect of this compound on protein synthesis. In order to determine the effect of AsIV on protein synthetic pathways, C2C12 murine muscle cells were treated with AsIV and the phosphorylation levels of key enzyme were assessed.

We found that AsIV induced phosphorylation of key protein synthetic targets in a time and dose‐dependent manner. By two hours of AsIV treatment, C2C12 cells had the greatest increase in AKT phosphorylation. These cells also had a 1.6‐fold increase in GSK‐3β phosphorylation and a 4‐fold increase in S6K1 activation, compared to controls. These results suggest that AsIV can phosphorylate AKT in a time‐dependent manner, with subsequent activation of S6K1 and inactivation of GSK‐3β. This suggests that AsIV may be capable of increasing the rate of protein synthesis in muscle cells. These findings help to further understand the molecular changes that occur in muscle cells and serve as a potential therapeutic mechanism to combat muscle loss.

Support or Funding Information

This work was supported by the Wehner Student Research Program at Montclair State University

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Immuno‐stimulating Activities of Purple Corn Stems
Authors: Kim Heeyeon , Park A‐Reum , Lee Hyo‐Young , Kwon Soon‐Bae
Published: June 2021

Maize hybrid grain, Saekso 1 was developed in 2008 by Gangwon Agricultural Research and Extension Services in Korea and registered in 2011. A single‐cross hybrid, Saekso 1 is a semi‐flint deep‐purple corn. Kernels are yellow, while the husks, cobs and stems are purple. Due to the Saekso 1 sensitivity to the excess moisture after seeding, water supply should be carefully managed, and it should be harvested at the suitable time to obtain the best anthocyanin pigment content. In this study, in order to enhance immune activities of Saekso 1 corn stems (S1S). Balb/c mice were feed daily for two weeks at S1S 1%, 10% and 15%. Treatment were to normal (general feed and saline), control (general feed and LPS) and treatment 1 (S1S 1% and LPS), treatment 2 (S1S 10% and LPS) and treatment 3 (S1S 15% and LPS). To measure body weight growth rate, the growth of splenocytes, the serum TNF‐α, IL‐1β, IL‐6 and IL‐10 level. The level of TNF‐α, IL‐1β, IL‐6 and IL‐10 production was detected by the ELISA assay when using the cytokine kit. Cytokine was statistically increased at supplemented groups with LPS and treatment with S1S significantly decreased the LPS‐induced TNF‐α and IL‐6 production. The results of this study may suggest that supplementation with S1S increases the immune function by regulating cytokine production capacity for activated macrophages. Further studies are needed to identify the stimulative and immunomodulating components of purple corn stems.

Support or Funding Information

Purple Corn

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Baclofen Does Not Modify the Gastric and Colonic Transits in Mice while It Inhibits Contractions of the Isolated Guinea Pig Ileum
Authors: Murad Hussam AS
Published: June 2021

Baclofen inhibits the transient lower esophag‐eal sphincter relaxations and thus it could be a potential treatment for the gastroesophageal reflux disease. The present work was conducted to study effects of baclofen on the gastrointestinal motility in experimental animals in three sets of experiments. In the first and second sets, 32 mice randomly divided into four groups (n=8) were used to evaluate effects of baclofen (4.0 mg/kg intraperitoneally) on the gastric transit (assessed by the charcoal meal test) and the colonic transit (assessed by the colonic bead expulsion time). In the third set, 16 guinea pigs randomly divided into two groups (n=8) were used to assess effects of baclofen (10 mM) on the electrically‐induced contractions of the isolated guinea pig ileum. Baclofen produced non‐‐significant changes in the gastric and colonic transits while it significantly inhibited the electrically‐induced contractions of the isolated guinea pig ileum. It is concluded that baclofen has favorable effects on the gastrointestinal motility supporting its potential value in treatment for the gastroesophageal reflux disease.

Support or Funding Information

None

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
High fat diet does not affect ADMA levels or ADMA regulatory enzymes in female Borderline Hypertensive Rats (BHR)
Authors: White Leslie Dallas , Mitra Anaya , Sasser Jennifer M , Wagner Laszlo , Rowland Neil E. , Baylis Chris
Published: June 2021

In this study, we tested the hypothesis that a HF diet increases ADMA levels in female BHR. Litters from male spontaneously hypertensive rats crossed with female Wistar rats that were maintained on either a HF diet (60% fat) or low fat diet (LF, 10% fat) for 6‐8 weeks prior to mating were used in this study. Offspring were divided into two groups: HHH (gestated in and suckled by a HF dam and weaned onto a "junk food" diet; n=6) and LLL (gestated in and suckled by a LF dam and weaned onto a standard chow diet; n=6). Rats were maintained on the respective diet for over 10 months, and then plasma levels of ADMA were measured by HPLC. HF diet increased body weight (499±32 vs 412±19 g, p<0.05 vs LF) but had no effect on blood pressure (118±7 vs 128±4, anesthetized direct arterial measurement). Analysis showed no significant changes in plasma ADMA levels (HHH: 0.18±0.02, LLL: 0.19±0.01 μmol/L). Kidney cortex abundance of the ADMA‐generating and degrading enzymes, protein methyltransferase‐1 (PRMT‐1) and dimethylarginine dimethylaminohydrolase (DDAH), respectively, and endothelial NOS (eNOS) were measured by Western blot. No differences in protein expression for any of the proteins were observed between groups (Table). Female BHR fed a HF diet did not express changes in plasma ADMA levels or DDAH, PRMT‐1 and eNOS expression.

(Western Blot data expressed as IOD/Ponceau red/Positive Control ± SEM)
Coronary Endothelial Ca2+‐Activated K+ Channels Under Hypoxic Exposure
Authors: Huang Jun‐Hao , Yang Qin , Underwood Malcolm John , Yao Xiao‐Qiang , He Guo‐Wei
Published: June 2021

Endothelial function is regulated by a number of ion channels, including intermediate and small conductance Ca2+‐activated K+ channels (IKCa and SKCa). Although endothelial dysfunction has been observed in hypoxic / ischemic states, little is known about the effect of hypoxia on IKCa and SKCa. Endothelial cells (ECs) were enzymatically isolated from porcine coronary arteries (PCA) and primary cultured for patch‐clamp study. Whole‐cell IKCa and SKCa currents were compared in ECs with or without hypoxic exposure (PO2 25‐40 mmHg, 10min). Vasorelaxant response to IKCa/SKCa activator 1‐EBIO (‐7~‐3.5 log M) was determined in small PCA (~1000 μm). Hypoxia markedly reduced whole‐cell K+ current (20.2±1.3 to 10.8±0.6 pA/pF) and the current activated by 1‐EBIO (30.3±2.8 to 21.4±1.7 pA/pF) (P<0.05). IKCa blocker TRAM‐34 significantly inhibited 1‐EBIO‐activation in both normoxic and hypoxic groups whereas SKCa blocker apamin exhibited relatively mild inhibition. Hypoxia reduced IKCa current from 10.5±2.2 to 7.3±1.7 pA/pF (P<0.05). Exposure to hypoxia for 1‐hr markedly decreased 1‐EBIO‐induced relaxation (87.0±6.9% to 67.8±7.2%, P<0.05). Endothelial whole‐cell currents consisting of IKCa and SKCa components are suppressed by hypoxia with marked inhibition on IKCa. This may contribute to the endothelium‐related impairment of dilatory function in coronary vasculature under hypoxic / ischemic states.
Relationship between Endogenous Fecal Zinc and Zinc Absorbed Revisited
Authors: Hambidge K Michael , Miller Leland V , Krebs Nancy F
Published: June 2021

Knowledge of the relationship between the quantity of endogenous zinc (Zn) excreted in the feces (EFZ, mg/d) and the quantity of exogenous Zn absorbed (TAZ, mg/d) at that time is necessary for estimating physiologic requirements for Zn. The objective of this project was to reevaluate this relationship utilizing all available total diet data (means), including new data since the publication of Dietary Reference Intakes for Zn. There were 30 datum including 11 male (M), 18 female (F) and 1 mixed gender. Results of linear regression analysis of EFZ vs TAZ and of TAZ at intercepta are summarized in the table:

aintercept of regression line for EFZ + other endogenous Zn losses versus TAZ with line of equality; bintercept based on using F other endogenous losses.

Lack of significant positive slope for F data is offset by higher y‐intercept resulting in similar intercepts for the F and combined data. The resulting difference in estimation of physiologic requirement for F is only 0.2 mg Zn/d, with actual figures depending on constants used for secondary routes of endogenous Zn excretion. The observed difference between M and F slopes and intercepts has no apparent physiologic explanation but could be attributable in part to limited range of TAZs for F data.
Dendrimer‐Based Nanomedicine: Its Impact on Biology, Pharma Delivery, MRI Imaging and Polyvalent Targeted Therapies
Authors: Tomalia Donald Andrew
Published: June 2021

In view of the importance of critical biological nanostructures such as: proteins, DNA/RNA, antibody‐antigen complexes, viral particles, etc., it is safe to make the following statement:

"The positive management of human health, disease and longevity will likely be determined and controlled by a deeper understanding of critical parameters in the nano‐length scale, namely: nanomedicine."

This theme will be used to present the use of precise, synthetic nanostructures (i.e., dendrimers/dendrons) as critical nanoscale building blocks in a variety of nano‐diagnostic, drug delivery and nano‐pharma applications [1‐2]. Dendrimers are now referred to as artificial proteins based on their close scaling and mimicry of protein dimensions, shapes and surface chemistries [3‐4]. Important features exhibited by dendrimers/dendrons such as nano‐scaffolding, nano‐container and nano‐chemistry features have led to the development of many well recognized commercial products (i.e., nano‐diagnostics (Stratus®, Siemens, Germany), nanoscale gene vectors‐ (Superfect®, Qiagen, Germany) and siRNA vectors (PrioFect®, Starpharma/Dendritic Nanotechnologies, Inc., Australia) [5]. Presently, a polyvalent dendrimer‐based nano‐pharmaceutical (VivaGel®, Starpharma, Australia) is in Phase II clinical trial development as an HIV/genital herpes microbicide. These successful developments portend the future evolution of many new dendrimer‐based nano‐devices/prototypes of importance to nanomedicine. Those possibilities will be overviewed in this lecture.
Skin Health and Metabolic Complications
Authors: Juturu Vijaya
Published: June 2021

Aging and metabolic complications (MC) such as diabetes mellitus (DM), hypertension, heart disease and hormonal changes were independent risk factors for dry and fissured skin. Rough, dry and scaly skin affects at least 75% of people with DM and other MC in aging. Intense itching (pruritus) is the most prominent in MC and enhance scratching and rubbing that aggravates dry skin condition, causing more inflammation and may end up in getting secondary infections. In MC, microvascular impairment may dehydrate the skin and cracks with erythema (redness or inflamation) and more evident in and around the involved areas in MC. Skin health plays a considerable role in managing skin conditions in chronic MC. Table 1 provides the MC and changes in skin health. It is very important to find the ways to rehydrate for healthy skin but also need to support the induction of proper epidermal differentiation.