bluuebunny/crossref_metadata_2025 huggingface dataset:
Leukocyte‐Specific Protein 1 (LSP1) regulates neutrophil migration in acute lung inflammation
Authors: Le Nguyen Phuong Khanh , Channabasappa Shankaramurthy , Liu Lixin , Singh Baljit
Published: June 2021

LSP1, an F‐actin binding protein, plays a role in neutrophil recruitment in peritoneum. Because mechanisms of excessive migration of activated neutrophils, credited with tissue damage, are not fully understood, we explored the hitherto unknown expression and role of LSP1 in neutrophil migration in acute lung inflammation (ALI). We induced ALI through intranasal E. coli LPS (80μg) in wild type 129/SVJ (WT) and LSP1 deficient (LSP−/− ) mice. WT (n=10) and LSP−/− (n=11) mice showed significant neutrophilia and more neutrophils in broncho‐alveolar lavage (BAL) at 9 hr post‐LPS challenge compared to respective saline‐treated controls (WT=7; LSP−/−=10). BAL neutrophil numbers were higher in LPS‐treated WT mice compared to LSP−/− mice (P<0.001). Lung myeloperoxidase and Gr1+ were higher in LPS‐treated WT compared to the LSP−/− mice (P<0.05). Lung tissue and BAL fluid KC, MCP1, MIP1α and MIP1β concentration and vascular permeability were not different between LPS‐treated WT and LSP−/− mice but TNFα concentration was higher in LPS‐treated WT mice. LSP1 expression was increased in inflamed lungs from LPS‐treated mice, and autopsied lungs from septic humans, compared to their respective controls. H&E staining showed more septal congestion in LPS‐treated WT mice compared to LSP−/− mice. These data show that LSP1 expression is modulated in ALI and that LSP1 deficiency reduces neutrophil migration into ALI. Funding: NSERC
Role of cytosolic glucocorticoid receptor and Annexin‐A1 on neutrophil traffic from bone marrow into blood: SDF‐ 1alpha/CXCR‐4/CXCR‐2 axis
Authors: Farsky Sandra Helena Poliselli , Machado Isabel Daufenback , Santin Jose Roberto , Ferraz Viviane Paula , Gil Cristiane Damas , Oliani Sonia Maria , Perretti Mauro
Published: June 2021

Aims

Investigate the role of the glucocorticoid cytosolic receptor (GCR) and ANXA1 on neutrophil traffic between bone marrow (BM) and blood (PB), focusing on SDF1alpha/CXCR‐4/CXCR2 axis.

Methods

BM perfusate and PB were collected from ANXA1‐ null (ANXA1−/−) and Balb/C wild‐type mice. Balb/C were pretreated with the GCR antagonist (RU38486; RU). Leukocyte numbers, CXCR‐2, CXCR‐4 and ANXA1 expressions, SDF‐1alpha and G‐CSF levels, chemotaxis, and phagocytosis of senescent neutrophils by BM macrophages were quantified.

Results

RU and ANXA1−/− presented accelerated granulocyte maturation in BM and neutrophilia, and ANXA1−/− showed increased number of mature granulocytes in BM. ANXA1 expression on BM neutrophils is reduced in RU. RU and ANXA1−/− presented increased CXCR‐4+ and reduced CXCR‐2+ cells in BM and PB, respectively. These cells presented reduced migration to SDF‐1alpha. BM SDF‐1‐alpha levels were reduced in ANXA1−/− and BM GCSF levels were reduced in RU and ANXA1−/−. BM macrophages collected from RU or ANXA1−/− phagocyted lesser number of senescent neutrophils.

Conclusion

EG, via GCR, and ANXA1 modulates the granulocyte maturation, neutrophil locomotion between BM/PB/BM and the phagocytosis of senescent neutrophils by BM macrophages.
The role of kinase and RGS activity of GRKs in regulating the dopaminergic signaling in hemiparkinsonian rats
Authors: Ahmed Mohamed Rafiuddin , Gurevich Vsevolod V , Gurevich Eugenia V
Published: June 2021

L‐DOPA is the treatment of choice for Parkinson's disease (PD) but prolonged exposure to the drug often leads to exaggerated signaling via dopamine (DA) receptors and motor complications such as L‐DOPA‐induced dyskinesia (LID). DA receptors are desensitized via activation‐dependent receptor phosphorylation by GRKs followed by the binding of arrestins, which stops further signal transduction. We previously reported that elevated expression of GRK6 in the striatum (Ahmed et al, 2010, Sci Trans Med, 2(28), 28ra28) ameliorated LID by promoting desensitization of DA receptors. Now we confirm that GRK6 acts via phosphorylation, since kinase‐dead protein mutant was without effect. Another GRK isoform, GRK3, potently inhibited L‐DOPA‐induced contralateral rotations in hemiparkinsonian rats. In contrast to GRK6, kinase‐dead GRK3 mutant suppressed rotations as effectively as wild type GRK3. This had prompted us to test the hypothesis that the GRK3 act via its RGS domain known to down regulate the signaling by sequestering Gaq/11. To address this question we used separate GRK3 RGS domain and RGS dead GRK3 constructs in the unilateral 6‐OHDA lesioned rat model. GRK3 RGS countered the rotational behavior, whereas the RGS‐dead GRK3 was ineffective. The data demonstrate that GRK3 regulates dopaminergic signaling and behavioral sensitization to L‐DOPA via its RGS domain.

Grant Support: NS45117 and NS065868 to EVG
Uridine enhances in vivo acetylcholine release in rat striatum
Authors: Cansev Mehmet , Yaylagul Esra Orenlili , Orhan Fulya , Turkyilmaz Mesut
Published: June 2021

The purpose of this study was to investigate the effects of parenteral uridine administration on acetylcholine release by in vivo microdialysis in striatum of freely‐moving rats.

Microdialysis probes inserted into left striatum of rats were perfused with artificial cerebrospinal fluid. After collection of 3 basal microdialysate samples, uridine was injected intraperitoneally. In additional experiments, P2Y receptor antagonism was achieved by intracerebroventricular injection of suramine 15 min prior to uridine administration. Levels of uridine nucleotides, choline and acetylcholine in microdialysate samples were analyzed by High Performance Liquid Chromatography.

Intraperitoneal administration of uridine enhanced brain extracellular uridine diphosphate (UDP), uridine triphosphate (UTP) levels and acetylcholine levels, while extracellular choline levels decreased after uridine. Suramine, a non‐selective P2Y receptor antagonist, blocked the alterations in extracellular acetylcholine and choline levels.

Our data provide the first in vivo evidence for the existence of pyrimidinergic transmission in the brain which can be enhanced by exogenous uridine administration. We also show for the first time that increased pyrimidinergic transmission enhances cholinergic neurotransmission, possibly by activating P2Y receptors.

This work was supported by grants from TUBITAK 108S354.
Chocolate may attenuate cognitive deterioration in Alzheimer's disease through prevention of gene expression related to depressive disorder
Authors: Pasinetti Giulio Maria
Published: June 2021

Chocolate has been shown to improve cognitive behavior. In particular, there is evidence that it may reduce dementia and attenuate cognitive deterioration in experimental models of Alzheimer's disease (AD) as well as in elderly patients with mild cognitive impairment. One condition associated with onset of experimental AD mouse models is characterized by depression and anxiety. A recent study suggests that the brain of a subject with major depressive disorder may be characterized by lower brain volume and major synaptic loss in humans. In addition, recent evidence suggests that experimental models of depression are characterized by decreased expression of downstream genes associated with transcriptional factor GATA1, which are genes associated with synaptic plasticity. Ongoing studies in our laboratory suggest that certain preparations of natural cocoa with enriched epicatechin levels may promote GATA1‐mediated transcription of genes associated with brain plasticity coincidental with promotion of long‐term potentiation. Our studies will clarify the role of chocolate epicatechin metabolites in the modulation of depression‐related genes in a mouse model of AD.
Exogenous T3 administration provides neuroprotection in a murine model of traumatic brain injury
Authors: Paterniti Irene , Crupi Rosalia , Campolo Michela , Di Paola Rosanna , Cuzzocrea Salvatore , Esposito Emanuela
Published: June 2021

Traumatic brain injury (TBI) induces primary and secondary damage in both the endothelium and the brain parenchyma that exacerbates the initial injury. Controlled cortical impact injury (CCI) is a widely‐used model of TBI. Here, we used a mice model of experimental TBI, to determine whether 3,5,3′‐triiodothyronine (T3) attenuates post‐traumatic neurodegeneration and neuroinflammation. Treatment with T3 (100 μg/kg, i.p.) 1 hour after TBI resulted in a significant improvement of motor and cognitive recovery after CCI, as well as markedly reducing lesion volumes. Mouse model for brain injury showed an increase of glial fibrillary acidic protein, a marker of a reactive astrocytes, and formation of inducible nitric oxide synthase (iNOS). Western blot analysis revealed the ability of T3 to reduce brain trauma through modulation of cytoplasmic‐nuclear shuttling of nuclear factor‐κB (NF‐κB).Twenty‐four hrs after brain trauma,T3‐treated mice also showed significantly less number of TUNEL(+) apoptotic neurons and curtailed induction of Bax, compared to vehicle control. In addition, T3 significantly enhanced the post‐TBI expression of the neuroprotective neurotrophins (BDNF, and GDNF) compared to vehicle. Our data provide an additional mechanism for the anti‐inflammatory effects of thyroid hormone with critical implications in immunopathology at the cross‐roads of the immune‐endocrine circuits.
Dopamine transporter affects cellular localization of α‐synuclein
Authors: Butler Brittany , Goodwin Shawn , Zarindast M , Mehr Shahram E , Azghadi Sara Hamidi , Khoshbouei Habibeh
Published: June 2021

The dysregulation of dopamine transporter (DAT) and over‐expression of α‐synuclein have been implicated in Parkinson's disease. We and others have shown a direct interaction between DAT and α‐synuclein. The objective of this study is to elucidate the localization of this interaction and to examine whether α‐synuclein over‐expression may affect the viability of dopaminergic neurons. We employed complementary approaches to examine the location of this interaction and how it may alter the activity of either protein in two model systems: heterologous expression system over‐expressing fluorescently labeled DAT and α‐synuclein as well as primary culture of mice DA neurons. α‐synuclein over‐expression in DAT cells decreases DAT‐mediated substrate uptake. Dialysis of α‐synuclein into DA neurons produced a DAT‐mediated, Cl‐dependent depolarizing current. To examine the locale of DAT/α‐synuclein interaction we used biochemistry and microscopy approaches including fluorescence recovery after photo‐bleaching (FRAP) and Forster resonance energy transfer (FRET). Biotinylation and FRET results suggest a direct DAT/α‐synuclein interaction at the plasma membrane. FRAP results suggest a transient interaction between the two proteins as indicated by a decrease in only α‐synuclein mobility. Ongoing experiments examine whether excess α‐synuclein influences the biology of dopaminergic neurons.
Prenatal hypoxia causes long‐term retina dysfunction in rats
Authors: Bourque Stephane Louis , Kuny Sharee , Sauve Yves , Davidge Sandra T
Published: June 2021

Background

Evidence indicates that prenatal stressors, such as hypoxia, can induce phenotypic changes in the fetus which increase its predisposition to age‐related complications. We hypothesized that aged offspring exposed to prenatal hypoxia would exhibit exacerbated age‐related functional decline of the retina.

Methods

Female Sprague‐Dawley rat offspring (that were maintained at either 11% O2 [hypoxic] or 21% O2 [normoxic] from gestational day (GD)15 to GD21, term=GD22) had their retina function assessed 2 and 14 months of age by electroretinophy (ERG). Anatomy was assessed by immunofluorescence microscopy.

Results

Dark‐adapted mixed a‐wave (rod and cone function) and b‐wave (bipolar cell activity) amplitudes were diminished in hypoxic offspring; these effects were most apparent at 14 months. Young and aged hypoxic offspring also had fewer mixed a‐wave threshold responses, and mixed b‐wave responses were markedly reduced in light adapted conditions. Light‐adapted flicker fusion frequency was also reduced in aged hypoxic versus age‐matched normoxic controls. There was no evidence of reduced photoreceptor cell numbers, although there was evidence of glial fibrillary acidic protein (GFAP) accumulation in aged IUGR offspring, providing evidence of photoreceptor damage.

Conclusions

Prenatal hypoxia causes both intrinsic and age‐related changes in retina function.
Mrp1 Protects Against Doxorubicin‐induced Cardiotoxicity in vitro
Authors: Zhang Wei , Coy Donna , Deng Jun , Vore Mary
Published: June 2021

Doxorubicin (DOX), an anthracycline antibiotic commonly used as a cancer chemotherapeutic agent, is well known for its ability to induce acute and chronic cardiotoxicity, leading to irreversible cardiomyopathy. In the present study, we investigated the role of multidrug resistance‐associated protein 1 (Mrp1), an ATP‐binding cassette efflux transporter, in DOX induced cardiotoxicity. We cultured cardiomyocytes (CM) and cardiac fibroblasts (CF), which were isolated from C57BL/6J (WT) or Mrp1 knock out (KO) mice (1 – 3 days old). The cells were treated with DOX (0.5 – 3 μM) and examined 24 hrs later. DOX enhanced Mrp1 mRNA (~2 fold) and protein (~2.6 fold) expression similarly in both CM and CF. The MTS assay showed that the CM from KO mice were more sensitive to DOX than WT, with an IC50 in KO of 0.94 μM (95% CI: 0.74 – 1.18) versus 1.60 μM (95% CI: 1.20 – 2.11) in WT. Furthermore, CM and CF from KO mice showed increased caspase 3 and PARP cleavage following DOX treatment, consistent with increased apoptosis in cells from KO. These results imply that Mrp1 protects against DOX induced cardiotoxicity in cultured CM and CF. (CA139844)
A herbal medicine decoction, Sibjotang, increases ANP secretion
Authors: Kwon Oh Jeong , Kim Hye Yoom , Kho Joung Hyun , Tan Rui , Cho Kyung Woo , Kang Dae Gill , Lee Ho Sub
Published: June 2021

Sibjotang (SJT), which was first documented in the Shanghanlun, has been prescribed for the remedy of body fluid disorders. The present study was designed to investigate the effect of SJT on Atrial natriuretic peptide (ANP) secretion, atrial dynamics and the possible mechanisms in beating rabbit left atria paced electrically. SJT significantly increased ANP secretion and atrial dynamics such as stroke volume and pulse pressure in a concentration‐dependent manner in perfused atria. EK or DG, components of SJT, also significantly increased ANP secretion and atrial dynamics, whereas, there were no significance on ANP secretion and atrial dynamics by treatment of EP or ZJ, respectively. Interestingly, the SJT‐induced increase of ANP secretion was significantly higher than the sum of each EK‐induced increase and DG‐induced increase of ANP secretion (p<0.01). SJT‐induced increase in ANP secretion was attenuated by GO6976, PKC ¥á/¥â inhibitor. In addition, LY333531, PKC ¥â inhibitor, also decreased SJT‐induced ANP secretion.

In conclusion, SJT‐induced ANP secretion was potentiated by combination with 4 components in beating rabbit atria. It provides an evidence for synergy effects of a herbal medicine decoction consisting of two or more components. Also, the data suggest that SJT‐induced ANP secretion was related with the activation of PKC ¥á/¥â signaling in perfused beating rabbit atria.
Neonatal Phthalate Exposure Induces Cardiac Dysfunction in Adolescent Rats
Authors: Yang Shu‐Han , Yang Rei‐Cheng , Wang Dean‐Chuan
Published: June 2021

Di‐(2‐ethylhexyl) phthalate (DEHP) is a widely used plasticizer found in a variety of polyvinyl chloride (PVC) products. Many evidences suggest that DEHP leached from PVC products may cause health problems in humans. Treatment of DEHP leads to arrhythmia in cultured cardiomyocytes suggesting that DEHP may act as an arrhythmogenic agent. Because infant babies and school‐age children are more vulnerable to environmentally toxic agents, there is a need to evaluate the effects of neonatal DEHP exposure‐induced cardiac dysfunction in adolescent rats. Sprague‐Dawley dams were orally fed with DEHP (10 mg/kg/day) or corn oil everyday during lactational period. At the age of 8 weeks, male pups were anesthetized and subjected to electrocardiogram (ECG) detection and then sacrificed for heart isolation. The proteins from left ventricle were extracted for Western blotting. The results show that bradycardia and prolonged QT intervals were observed in DEHP group. Meanwhile, the expressions of heat shock protein 70 (HSP70), a biomarker of protective mechanism in myocardium, were significantly decreased in DEHP group. These results indicate that DEHP exposure in the early life stage may cause cardiac dysfunction by decreased HSP70 protection in adolescent animals.
Effects of Reduced Uterine Perfusion Pressure on Cerebral Artery Tone in Pregnant Rats
Authors: Dawson Jack , Kilar Cody , Novak Jacqueline , Ramirez Rolando J.J.
Published: June 2021

Preeclampsia is the leading cause of maternal and fetal mortality and it occurs in 5–10% of pregnancies. The etiology of the disease is not well understood but reductions in placental perfusion are central to disease. In patients with preeclampsia (PE) and rodent models of PE alterations in vascular reactivity are indicated. However, the vascular reactivity of the cerebral circulation has not been well characterized, despite the fact that preeclampsia may lead to seizures (eclampsia). We hypothesize that reduced uterine perfusion pressure (RUPP) in pregnant rats, a well accepted model of preeclampsia pathophysiology, will increase vascular reactivity of cerebral arteries isolated from RUPP pregnant rats. For this study, pregnant Sprague Dawley rats were subjected to a surgery to reduce uterine perfusion pressure or a sham operation at gestational day 14. At gestational day 20, cerebral arteries were isolated and studied in an isobaric arteriograph. Active and passive diameters were measured at pressure steps ranging from 25 to 200 mmHg. Our results showed an increase in tone in cerebral arteries of RUPP compared to sham controls (i.e. 12% vs. 6% at 125mmHg). These results support our hypothesis that RUPP surgery in rats can alter cerebral vascular reactivity and allow us to explore the mechanism which may predispose the PE patients to seizures and strokes.
GLYOXAL AND METHYLGLYOXAL BUT NOT 4‐HYDROXYNONENAL AND MALONDIALDEHYDE FORM ADDUCTS ON CARDIAC RYANODINE RECEPTOR (RyR2) AND SARCO(ENDO)PLASMIC RETICULUM Ca2+ ATPASE (SERCA2) IN DIABETES
Authors: Bidasee Keshore , Alomar Fadhel , Singh Jaipaul , Moore Caronda
Published: June 2021

Posttranslational modification of cardiac type 2 ryanodine receptor (RyR2) and sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA2) by reactive carbonyl species (RCS) is emerging as a leading cause for disturbances in the orchestrated release of Ca2+ from and reuptake into the sarcoplasmic reticulum (SR) in diabetes. What is less clear at this time is which of the many RCS that are upregulated in diabetes chemically modify and impair the functions of RyR2 and SERCA2. This study uses in vitro assays to delineate the impact of key RCS that are upregulated during diabetes glyoxal (GO), methylglyoxal (MGO) and 4‐hydrononenal (4‐HNE) on functions of RyR2 and SERCA2. It also used adduct‐specific antibodies to determine if GO, MGO, 4HNE and malondialdehyde (MDA) formed adducts on RyR2 and SERCA2 in a rat model of type 1 diabetes. In [3H]ryanodine binding and lipid bilayer assays, MGO, GO and 4HNE binded to and modulated the open probability (Po) and/or conductance of RyR2 with varying potencies, establishing that they are capable of interacting with RyR2. In Ca2+ uptake assays, MGO, GO and 4‐HNE also dose‐dependently modulated the ability of SERCA2 to transport Ca2+. Interestingly, RyR2 and SERCA2 isolated from hearts of rats with diabetic cardiomyopathy contained MGO and GO but not 4‐HNE MDA adducts. From these data that although MGO, GO and 4‐HNE are capable of altering the function of RyR2 and SERCA2 in vitro assays, only glucose‐derived MGO and GO preferentially react with RyR2 and SERCA2 to form adducts. Reasons for this selectivity will be discussed. Supported in part by NIH HL085061
Specialized role of Ca2+‐activated K+ channels in signaling responses of different brain cell types
Authors: Gebremedhin Debebe
Published: June 2021

The change in membrane electrical properties is a critical determinant of cellular function. Potassium channels are ubiquitous native membrane proteins and play a role in a variety of cellular functions induced by endogenous or exogenous stimuli. The KCa channels are unique members of voltage‐gated K+ channel family in that they are activated both by voltage and a rise in [Ca2+]i. The KCa channels are important regulators the resting membrane potential and link extracellular signals with intracellular second messenger systems. The KCa channels expressed in astrocytes and neurons are different from those native in cerebral arterial muscle cells (CAMCs) in that they are insensitive to the toxin blockers of KCa channel due to expression of the KCa channel fÒ4 subunit. Blockade of KCa channels results in membrane potential depolarization that permit Ca2+ influx in CAMCs and neurons but not in astrocytes, whereas activation of KCa channels causes membrane potential hyperpolarization in all cell types that inhibits Ca2+ influx in VSMCs and neurons, but facilitates Ca2+ influx in astrocytes that is crucial for neurovascular coupling.
Regeneration of injured soleus muscle in heat shock transcription factor 1‐null mice
Authors: Goto Katsumasa , Nishizawa Sono , Koya Tomoyuki , Nakai Akira , Sugiura Takao , Ohira Yoshinobu , Yoshioka Toshitada
Published: June 2021

Heat shock transcription factor 1 (HSF1) mediates stress response, and up‐regulates heat shock proteins (HSPs) in mammalian skeletal muscles. The purpose of this study was to investigate the effects of HSF1‐deficiency on the regeneration of inured skeletal muscle. Cardiotoxin was injected into the left soleus muscle of mice. Soleus muscles were dissected bilaterally 2 and 4 weeks after the treatment. HSF1‐deffiency retarded both the recovery of muscle protein content and fiber size, which were decreased by cardiotoxin injection. Injury‐related‐up‐regulation of HSPs in HSF1‐null mice was depressed, compared with wild type mice. HSF1‐associated stress response may play a key role in the regeneration of injured skeletal muscle. This study was supported, in part, by KAKENHI (22240071, 24650411, 24650407) from Japan Society for the Promotion of Science and the Science Research Promotion and the Promotion and Mutual Aid Corporation for Private Schools of Japan.
Insulin sensitivity and pancreatic function in type 2 diabetics with and without insulin treatment
Authors: Gavin Timothy P , Ernst Jacob M , Kehe Sarah E , Dohm G Lynis , Pories Walter J , Dar Moahad , Reed Melissa A
Published: June 2021

As type 2 diabetes (T2D) progresses, often the addition of insulin is required along with oral glycemic medications for glycemic control. We questioned if pancreatic function, insulin sensitivity, or both were lower in T2D using insulin (INS: N=9) vs. oral glycemic agents alone (OGA: N=12). Hemoglobin A1c (HbA1c) was measured. An insulin modified intravenous glucose tolerance test (modified‐IVGTT) was performed and the acute insulin response to glucose (AIRg) and insulin sensitivity (SI) were calculated using minimal model analysis. HbA1c (INS: 8.1±0.5 %; OGA: 6.5±0.2 %; p < 0.01) was higher in INS vs. OGA. AIRg (INS: 13±10 μU/ml; OGA: 131±45 μU/ml; p = 0.04) and SI (INS: 0.6±0.2; OGA: 1.9±0.3; p < 0.01) were lower in INS vs. OGA. Multiple stepwise linear regression revealed that SI explained 47% and AIRg 14% of the variance in HbA1c. These results suggest that both pancreatic function and insulin sensitivity are lower in T2D patients whose medications include insulin compared to patients on oral glycemic agents alone and that differences in insulin sensitivity explain more of the variance in glycemic control than differences in pancreatic function in type 2 diabetics.

Supported by an East Carolina Diabetes and Obesity Institute (ECDOI) grant.
Low RIP140 expression rescues insulin‐mediated FA oxidation via a rise in MCAD gene expression in skeletal muscle cells exposed to high palmitate availability
Authors: Constantinescu Silvana , Turcotte Lorraine P
Published: June 2021

We have shown that in L6 cells incubated with high palmitate, low Receptor Interacting Protein 140 (RIP140) expression alters the regulation of muscle fatty acid (FA) uptake and FA oxidation under insulin‐mediated conditions. To provide mechanistic insights for these metabolic changes, we measured mRNA and/or protein expression of several genes involved in the regulation of FA uptake and oxidation. Cells were incubated with insulin (1000 nM) ± palmitate (400 μM) and with siRNA sequences for either RIP140 or a negative control. Under insulin‐mediated conditions, short‐term incubation with palmitate decreased (P<0.05) FA uptake (24%) and oxidation (42%). Under these conditions, low RIP140 expression reduced (P<0.05) FA uptake (35%) and restored FA oxidation so that it was not different (P>;0.05) from the control rate. Incubation with palmitate increased (P<0.05) CD36 (70%) and FATP1 (224%) mRNA, decreased (P<0.05) CPT1 (45%) mRNA and did not change MCAD (P>;0.05) mRNA. High palmitate was accompanied by reductions (P<0.05) in protein content for FATP1 (68%) and total (59%) and plasma membrane (31%) CD36 but it did not affect (P>;0.05) CPT1. Under those same conditions, low RIP140 expression was accompanied by a rise in the mRNA expression of MCAD (83%; P<0.05) and FATP1 (38%; P=0.08) but no change (P>;0.05) in mRNA expression of CPT1 and CD36. Protein content of CPT1, FATP1, CD36 (total and plasma membrane) were not affected (P>;0.05) by low RIP140 expression. Our results indicate that low RIP140 expression in the context of high palmitate availability increases gene expression of MCAD and that this may in part explain the restoration of insulin‐mediated FA oxidation.

USC Women in Science and Engineering, USC Integrative and Evolutionary Biology and USC Zumberge Research Innovation Fund
Body mass index and ethnicity as determinants of brown adipose tissue function as measured by thermal imaging in healthy children
Authors: Budge Helen , Elvidge Lindsay , Ojha Shalini , Symonds Michael E
Published: June 2021

Brown adipose tissue (BAT) function is greatest in children but the main factors influencing it remain to be determined. Our study was, therefore, designed to determine whether body mass index (BMI) centile influenced skin temperature overlying BAT depots in the supraclavicular region (SCR) of healthy children. Infrared thermography was used to assess SCR skin temperature at baseline and following exposure to a mild cool stimulus (single hand immersion in water at 20.1°C) over a period of 5 minutes in young children aged 6–11 years (n=55). There was a clear difference in SCR temperature between age matched healthy subjects who were either of low, or high, BMI. The primary indicator of baseline and stimulated SCR temperature was BMI centile for which there was an inverse relationship (R2 = 0.26; P = 0.009). Ethnicity was also a significant predictor of SCR temperature. We have demonstrated an inverse relationship between the temperature of the SCR, co‐locating with the primary region of BAT, and BMI centile. Future studies aimed at determining the primary factors regulating BAT function in healthy children could be used to prevent excess white adipose tissue deposition in early life.
Post‐Metabolic Response to Exercise in Normobaric Hypoxia
Authors: Kelly Liam Patrick , Basset Fabien Andre
Published: June 2021

The purpose of the study was to examine post‐metabolic response to constant workload exercise (CWE) under normobaric hypoxia. Seven active healthy males (age: 26±4 yrs; height: 176±4 cm; weight: 78±10 kg; BMI: 25±3 kg•m−2; VO2max: 4.0±0.3 L•min−1) randomly underwent normoxic and hypoxic (FiO2= 0.15) CWE. Each experimental session consisted of a basal metabolic rate determination (BMR), 60‐min of CWE, and 60‐min of post‐exercise metabolic rate measurement. For each condition, 24‐hr post‐treatment BMR was recorded. Absolute workload was determined during a graded cycling test in normoxia and set at 50% of participants’ peak power (314±26 W) for both conditions. There was a significant condition/time (0–20; 21–40, and 41–60‐min) interaction on post‐exercise lipid metabolism (p= 0.015). Fat contribution was 47±19%, 60±9% and 51±12% in normoxia; and 79±20%, 79±19% and 62±24% in hypoxia, for each epoch, respectively. Although not statistically significant, fat contribution 24‐hrs after the hypoxic intervention was elevated 3% above baseline. In conclusion, exercise in hypoxia contributed to an increase reliance on fat oxidation post‐treatment up to 24‐hrs. This result was explained by higher glycolytic flux during exercise under hypoxia compared to normoxia at the same absolute workload.
Using D2O to measure plasma protein synthesis after a single meal
Authors: Wagner David A. , Bolt Daniel , Greulich Knut
Published: June 2021

Our objective was to quantify plasma protein synthesis after a single meal with a bolus administration of deuterium oxide. Labeled body water rapidly equilibrates with amino acids such as alanine prior to incorporation into newly synthesized proteins. We utilized a gas chromatograph combustion isotope ratio mass spectrometer (GCC‐IRMS) to measure very low levels of deuterium isotope incorporation with high precision. We compared the rate of plasma protein synthesis in CD‐1 mice in four groups: water treated control or three amounts of whey protein (low, medium and high). After food deprived overnight, all mice received an intraperitoneal injection of deuterium oxide four hours prior to treatment. Enrichment of plasma body water, free alanine and alanine bound to plasma proteins was determined at 5–8 hours after control or protein treatment. The fractional rate of plasma protein synthesis was 17.5%, 29.6%, 37.5% and 43.9% for control, low, medium and high protein treatment respectively. These finding show that the GCC‐IRMS with deuterium labeling of body water can be used to detect differences in protein synthesis after a single meal.
Female sex steroid effects on basal muscle protein synthesis rates in postmenopausal women
Authors: Smith Gordon I , Reeds Dominic N , Burrows Rachel E , Kampelman Janine A , Mittendorfer Bettina
Published: June 2021

We recently reported a greater basal, postabsorptive rate of muscle protein synthesis (MPS) in post‐compared with premenopausal women. This finding suggests that ovarian hormones suppress MPS. In the current study, we tested this hypothesis by measuring the basal rate of MPS (by using stable isotope‐labeled tracer methods) in postmenopausal women before and after treatment with either estradiol (Mylan Pharmaceuticals; 0.1 mg/d delivered transdermally; n=6) or progesterone (Endometrin®; 100 mg/d delivered vaginally; n =5). The MPS rate was unaffected by estrogen treatment (0.063±0.006 vs. 0.063±0.010 %/h, before and after treatment respectively; P=0.99) but was markedly increased after progesterone treatment (0.102±0.018 %/h vs. 0.055±0.007; P=0.02). We conclude that the menopause associated decline in ovarian female sex hormone production is not responsible for the age‐associated increase in MPS in old women.

Supported by grants from the NIH.
Mouse embryonic fibroblasts devoid of both mitofusin1 and mitofusin2 can acutely shift from glycolytic to oxidative metabolism.
Authors: Seifert Erin , Moffat Cynthia , Strus Ewa
Published: June 2021

The extent to which a deficiency of mitochondrial fusion is required for oxidative metabolism is unclear. To this end, we have undertaken a bioenergetics analysis using intact mouse embryonic fibroblasts deficient in mitofusin1 and mitofusin2 (DKO) incubated with glycolytic vs. oxidative substrate. We first noted a ~doubling of non‐mitochondrial oxygen consumption (JO2) in DKO cells, regardless of substrate. Upon switching from glycolytic to oxidative conditions, both Wt and DKO MEFs increased basal mitochondrial JO2 (oligomycin‐sensitive+oligomycin‐insensitive, non‐mitochondrial subtracted), although the rise was smaller in DKO cells (increase of 55±11 vs. 114±11 pmoles/min/40k cells in DKO vs. Wt; mean±sem). Considering only oligomycin‐sensitive JO2, the rise was similar in DKO cells (increase of 121.3±10 vs. 135.8±13 pmoles O2/min/40k cells in DKO vs. Wt), whereas oligomycin‐insensitive JO2 decreased in DKO cells only (by ~50%). In both Wt and DKO cells, the rate of media acidification declined upon switching from glycolytic to oxidative substrate, indicating a shift from glycolytic to oxidative ATP production. Thus, as in Wt MEFs, DKO MEFs demonstrate basal mitochondrial respiration controlled by ATP turnover, and substrate oxidation. Differently from the Wt, DKO mitochondria exert significant control over basal JO2 by proton leak, which would function to expand capacity.
A systems biology analysis of the genomics of probenecid action in the late gestation ovine fetal pituitary
Authors: Wood Charles , Rabaglino Maria B , Chang Eileen , Richards Elaine
Published: June 2021

To understand the genomic response to transporter blockade in the fetal pituitary, we used chronically catheterized twin fetal sheep (123–133 days gestation) infused with probenecid (PBN) or vehicle for 5 days intracerebroventricularly (icv). At the end of the infusion, mRNA was extracted from pituitary and genomic expression analyzed using Agilent ovine arrays. PBN differentially regulated 155 genes (80 upregulated and 75 downregulated genes). Network inference revealed significant networks for up‐ and down‐regulated genes; however, because of the relatively small number of genes differentially regulated by the PBN, both up and downregulated genes were subjected to gene ontology analysis. Statistically significant biological processes included: response to nutrient; response to steroid; steroid biosynthetic process; and regulation of fibroblast differentiation. Hierarchy analysis of the DRG revealed that the gene highest in the hierarchy was ACTR3 (actin‐related protein 3, involved in cellular remodeling, shape, motility). ACTR3 is regulated by PANX1 (Pannexin‐1, gene is not on the array), an ion channel that is inhibited by PBN. We propose that the genomic response to PBN in the ovine fetal pituitary could be mediated entirely by the blockade of Pannexin‐1.
Norepinephrine reconfigures post‐inspiratory neurons within the pre‐Bötzinger complex of mice
Authors: Garcia Alfredo , Khuu Maggie Ann , Ramirez Jan Marino
Published: June 2021

The contribution of inspiratory neurons in central inspiratory rhythmogenesis is well established. However, the inspiratory neuronal phenotype is only one of several known to be present in the ventral respiratory column and the pre‐Bötzinger complex (preBötC), in particular. Here we characterized post‐inspiratory neurons (post‐In) in rhythmic transverse brainstem slices (550–620μm, mice P2–P12). Pharmacological and electrophysiological characterizations reveal that post‐In receive concurrent excitatory and inhibitory synaptic input in‐phase with the inspiratory population rhythm in the preBötC. This concurrent input allows post‐In to become conditional inspiratory neurons (In) in the presence of decreased synaptic inhibition. We also find that elevated noradrenergic tone (NE) transforms the firing patterns of some post‐In into conditional In. Optogenetic experiments made in ChAT‐positive neurons expressing channelrhodopsin 2 revealed that ChAT‐positive neurons with the post‐In phenotype are not transformed into In by NE, suggesting that it is non‐cholinergic preBötC interneurons that are characterized by this conditional phenotype. Our findings illustrate that concurrent inhibition and excitation characterizes post‐In in the preBötC and that the post‐In phenotype can be reconfigured by the presence of elevated NE.
Impact of Food Label Indications on Cooking Common Recipes: An Experimental Study on Chilean Mothers.
Authors: Gregori Dario , Vecchio Maria Gabriella , Ballali Simonetta , N2C Study Group
Published: June 2021

The portion size of food and drink products is an important consideration when providing nutrition information for labeling purposes. No data are available on how food label indications translate into real cooking strategies. An experimental trial was performed on Chileans mothers and a relative. Participants were asked to prepare two meals with ingredients measured only with kitchen tools. Participants were randomized to two groups: one receiving information using portion size (PSG) indications and the other using “per 100 gr” (100G) indications. Subsequently, participants were interviewed with a questionnaire assessing nutritional and portioning knowledge. Actual usage of ingredients was assessed using a high‐precision scale.

117 mothers and grandmothers participated. Oil usage was the same in both groups (p=0.124), for chapalele, about 4.25 g (2.975–6.481) in the 100G group and 5.56g (3.05–9.20) in the PSG group. For pasta, oil usage was 4.88g (3.08–7.75) in the 100G group and 5.35g (3.74–9.94), not resulting significantly different (p=0.425).

Food label indication does not seem to influence attitudes toward the amount used for basic ingredients in cooking common traditional recipies.
Fundamentals of Team‐based Learning and How it Can Work in Teaching Anatomy
Authors: Nieder Gary L.
Published: June 2021

Team‐based learning (TBL) is a well‐described instructional strategy, involving a sequence of individual and group learning activities, which promotes individual and group accountability, development of problem solving proficiency and improved teamwork skills. The core elements of TBL are: 1) individual preparation guided by specific assignments and objectives; 2) individual readiness assurance testing; 3) group readiness assurance testing; 4) group application exercises; 5) peer evaluation. Departure from this sequence is possible, but should be carefully undertaken and assessed to preserve all of the benefits of the method. TBL can be implemented alongside traditional anatomy learning activities such as readings, lecture and dissection lab or in alternative anatomy teaching situations. We have structured our TBL sessions to deal with the highly visual nature of the anatomical sciences, including the use of laptops and now iPads to provide each team with high quality radiological imaging, figures and video. We also include team‐produced drawings, with subsequent peer evaluation, as part of the application problems in most of the TBL sessions. TBL has proven to be a valued addition to the overall anatomy and embryology teaching program.
Human Cardiovascular Progenitor Cells Positive for Flt1 and Flt4 Surface Cell Markers Differentiate Into All Three Cell Types of the Cardiovascular Lineage
Authors: Nsair Ali
Published: June 2021

Introduction

Heart disease remains a leading cause of morbidity and mortality. One of the major limitations in cardiovascular cell therapies are the poorly characterized cell types used so far, which have limited capacity for robust cardiac differentiation.

Purpose of study

Surface markers that characterize cardiac progenitor cells (CPCs) are not well‐defined. To identify cell surface markers to identify this population of cells, we performed a microarray analysis on CPCs isolated from mouse cells. Combinatorial testing of candidate cells surface marker revealed that a combination of Flt1 and Flt4 best identified CPCs.

Results

Flt1+/Flt4+ progenitors in human fetal hearts demonstrated expression of CPC‐specific genes but no markers of differentiated cardiovascular cells. Flt1+/Flt4+ population from human pluripotent cells developed spontaneously beating colonies with cardiomyoctyes and smooth muscle and endothelial cells.

Conclusion

The capability of pluripotent stem cell‐derived human Flt1+/Flt4+ cells to differentiate into all three cell types of the cardiovascular lineage suggests that this progenitor cell might be a good candidate for regenerative therapies.
Impact of national policy banning TV advertisement on High‐energy/Low‐nutrient foods
Authors: Lee Soo‐Kyung , Nam So‐Young , Yoon Barbara Jihyun , Chung Sang‐Jin
Published: June 2021

High‐energy/low‐nutrient (HELN) food advertizing targeting children has been identified as a contributing factor to child obesity. Korea has enacted a national policy banning TV advertisement of HELN foods from 5pm to 7pm in 2010, aiming to control child obesity. This study examined how the HELN food consumption and obesity have been affected by the policy, using the Korea National Health and Nutrition Examination Survey from 2008 to 2010. The 24‐hr recall data of children (2–18 year old) were grouped into two: 1) children who consumed at least one HELN food on the day of recall and 2) children who did not consume any HELN foods. Before the national policy was enacted, 18% of children reported having a minimum of one HELN food, while 21% reported in 2010 after the policy enacted. The number of HELN foods consumed also appeared to increase after the policy. Interestingly, the proportion of HELN foods against total food consumed decreased from 19.06% to 16.65%, and the decrease was more significant among obese children. No significant changes were observed in child obesity status. This finding would be useful to designing advertising‐related food policies. This research was supported by a grant (12162‐KFDA‐110) from Korea Food and Drug Administration.
Decaffeinated green tea extract and voluntary exercise alters the expression of genes related to lipid metabolism and mitochondrial biogenesis in liver and skeletal muscle in high fat‐fed mice
Authors: Sae‐tan Sudathip , Rogers Connie J , Lambert Joshua D
Published: June 2021

Obesity is a significant health problem. Studies have reported the beneficial effects of green tea for weight loss and exercise for weight management. We have previously reported the combination of decaffeinated green tea extract (GTE) and voluntary exercise (Ex) has greater effects on high fat diet‐induced obesity and comorbidities in male C57BL/6J mice than either treatment alone. Here, we report that the combination alters gene expression in the liver and skeletal muscle. mRNA levels of sterol regulatory element‐binding protein‐1c and sterol‐coenzyme A desaturase‐1, genes related to fatty acid synthesis, were increased by 1.4‐ and decreased by 0.5‐fold, respectively, by GTE+Ex treatment in the liver. Hepatic mRNA expression of peroxisome proliferatoractivated receptor‐α and carnitine palmitoyltransferase‐1α, genes related to fatty acid oxidation, were increased by 2.2‐ and 1.3‐ fold, respectively, by GTE+Ex treatment. GTE+Ex treatment also increased the mRNA expression of proliferator‐activated receptor‐ γ coactivator −1α, cytochrome b, cytochrome C oxidase subunit III by 1.5‐, 1.5‐, and 1.3‐fold, respectively, in the skeletal muscle. Our results suggest that the combination of GTE and Ex modulates the expression of several genes related to lipid metabolism and mitochondrial function. These changes may underlie the obesity preventive effects of this combination.

Grant Funding Source: NIH Grant (AT004678)
Parental Recognition and Perception of Commercially Available Foods and Beverages Containing Non‐nutritive Sweeteners
Authors: Sylvetsky Allison Cara , Greenberg Mitchell J , Rother Kristina I
Published: June 2021

Objective

Our study aimed to examine parental attitudes toward providing NNS containing items to their children and to explore parental ability to recognize NNS in packaged foods and beverages.

Methods

120 parents (≥ 18 years old) of children (≥1 year old) completed brief questionnaires upon entering or exiting a local grocery store. Grocery selection and NNS recognition were assessed using a 142 item grocery shopping simulation activity. Demographic information and parental attitudes toward NNS were assessed using an interviewer‐assisted survey.

Results

A factor analysis generated 3 subscales of from the NNS attitude questionnaire; nutrition and health awareness (NHA), NNS acceptance (NNS‐ACCEPT), and child sweet liking (SL). Overall our findings demonstrated that the majority of parents (72.3%) did not believe that NNS were safe to provide to their children, yet overall ability to recognize NNS was low (mean percentage of items correctly = 22.9%). Accurate parental identification of NNS containing foods and beverages was associated with heightened negative attitudes towards provision of NNS to their children.

Conclusion

Future well‐designed studies must be conducted to assess the short‐ and long‐term metabolic effects of NNS in order to formulate conclusive recommendations promoting the use or avoidance of NNS. Nutrition education promoting ingredient list and nutrition facts panel comprehension, as well as food claim literacy is required to reduce misguided nutrition beliefs among parents.
MINOCYCLINE BLOCKS ALLERGEN‐INDUCED EOSINOPHILIA AND PRODUCTION OF TH2 CYTOKINES AND IGE BY INTERFERING WITH THE T CELL RECEPTOR‐NF‐kB‐GATA‐3‐INTERLEUKIN (IL)‐4 AXIS IN A MURINE ASTHMA MODEL WITHOUT AN EFFECT ON PARP
Authors: Naura Amarjit S , Kim Hogyoung , Ju Jihang , Rodriguez Paulo C , Jordan Joaquin , Pyakurel Kusma , Tarhuni Abdelmetalab F , Ochoa Augusto C , Boulares Hamid A
Published: June 2021

Minocycline protects against asthma independently of its antibiotic function. Minocycline was recently reported as a potent PARP inhibitor. In an animal model of asthma, a single administration of minocycline conferred excellent protection against ovalbumin (OVA)‐induced airway eosinophilia, production of mucus, Th2 cytokine and OVA‐specific IgE. These effects were unrelated to a potent inhibition of PARP; the drug moderately reduced PARP activation by modulating oxidative stress‐associated DNA damage. Minocycline failed to efficiently block direct PARP activation in lungs of MNNG‐treated mice or H2O2‐treated cells. Minocycline blocked OVA‐specific IgE production in B cells potentially by modulating T cell‐receptor (TCR)‐linked production at IL‐4 mRNA level but not through a modulation of the IL‐4‐JAK‐STAT‐6 axis, IL‐2 production, or NFAT‐1 activation. Such IL‐4 production blockade was linked to a preferential inhibition of the NF‐κB activation arm of TCR but not GSK3, Src, p38 MAPK, or ERK1/2. Inhibition of NF‐κB activation was linked to a complete blockade of TCR‐stimulated GATA‐3 expression, a pivotal transcription factors for IL‐4 expression. These results show that minocycline inhibits IgE production and asthma traits by modulating TCR function particularly by inhibiting the signaling pathway leading to NF‐κB activation, GATA‐3 expression, and subsequent IL‐4 production.
Embryonic origin of the skull: Has the pattern of neural crest derivation changed during vertebrate evolution?
Authors: Hanken James , Piekarski Nadine
Published: June 2021

The vertebrate skull is derived from embryonic neural crest and mesoderm. These contributions have been revealed in great detail in one amniote model and to a lesser extent in other taxa. The degree to which patterns of derivation are evolutionary conserved or labile among most lineages nevertheless remains largely unknown. Amphibians are a non‐amniote tetrapod group with a highly derived skull morphology and ontogeny. We employed embryonic transplantations, using GFP‐transgenic axolotls as donors, to document the nature and extent of neural‐crest contribution to the adult osteocranium in this species. Comparisons with chicken and mouse reveal a highly conserved embryonic origin of the tetrapod skull. Conversely, a comparison between axolotl and Xenopus reveals tremendous differences in the origin of skull bones between these two taxa. The unique features of Xenopus may be a consequence of the extensive, posthatching cranial metamorphosis that is characteristic of most anurans. The degree to which the embryonic origin of the vertebrate skull is conserved or labile varies according to lineage and is thus subject to evolutionary change.