bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mechanisms regulating the dopamine D1‐D2 receptor heteromeric signaling complex
Authors: Verma Vaneeta , O'dowd Brian , George Susan 
Published: June 2021

We previously identified that co‐activation of a Gq‐linked heteromeric D1‐D2 dopamine receptor signaling complex results in phospholipase C (PLC)‐dependent intracellular Ca2+ release. We have demonstrated agonist specificity for the D1‐D2 triggered Ca2+ signal and the D1 stimulated cyclic AMP production with three D1 agonists, SKF83959, SKF81297, and SKF83822 that differentially activate either the adenylate cyclase (AC) or PLC pathway. Because the three agonists can activate different signaling pathways, we propose that they are effective pharmacological tools for assessing the mechanisms regulating the D1‐D2 signaling complex. Pretreatment of D1‐D2 expressing cells with each agonist lead to 60% desensitization of the Ca2+ signal within 5 minutes. Intriguingly, occupancy of the D1 binding pocket by SKF 83822 that could not activate the D1‐D2 complex still permitted desensitization of the Ca2+ response. Results from co‐application of the agonists with a D1 or D2 specific antagonist, suggest the desensitization elicited by each agonist is through selective occupancy of the D1 receptor. Furthermore, over‐expression of wild type G‐protein receptor kinase 2 (GRK2) and its catalytically inert construct, but not its regulator of G‐protein signaling (RGS) domain impaired construct, decreased the Ca2+ signal, suggesting the RGS domain of GRK2 is involved in regulating the D1‐D2 receptor signaling cascade.This work is supported by the NIH National Institute on Drug Abuse
Cellular substrates for angiotensin1‐7 (Ang1‐7) action in the rostral ventro‐lateral medulla (RVLM) of the normotentsive and spontaneously hypertensive rat (SHR)
Authors: Guo Fang , Souslova Ekaterina , Chudakov Dmitry , Paton Julian F.R. , Kasparov Sergey 
Published: June 2021

Ang1‐7 is an important component of the central renin‐angiotensin system and affects haemodynamics when injected into the RVLM. The cellular substrate(s) that Ang1‐7 targets in RVLM is unknown., We studied the effects of Ang1‐7 in organotypic brainstem slice cultures of Wistar rat (WR) and SHR. Using cell‐selective promoters incorporated into adeno viral vectors, we expressed green fluorescent protein in catecholaminergic (CA‐) neurones and non‐CA neurones and a Ca2+ indicator Case12(1) in astroglia. . Ang1‐7 (200 nM) depolarised non‐CA neurones in both WR and SHR (by 5 and 4 mV, correspondingly; p<0.05), which in SHR, but not WR, was accompanied by an increase in [Ca2+]i of 61 ± 28% (P<0.05). In contrast, Ang 1‐7 (200 nM) had no measurable effect on either membrane potential or [Ca2+ ]i in CA neurones of either SHR or WR. In astrocytes Ang1‐7 also triggered increases in [Ca2+ ]i: 200nM Ang1‐7 evoked increases of 62±23% in [Ca2+ ]i in 26% of WR astrocytes whereas 600 nM Ang1‐7 activated more astrocytes (54%) but the average amplitude of response was similar. In SHR, Ang1‐7 effects were also similar. Effect of Ang1‐7 could be blocked by co‐application of its antagonist, A779 (200 nM) but not losartan (1 μM). We conclude that non‐CA neurones and glia in RVLM are the primary targets for Ang1‐7 and its effect on [Ca2+]i in non‐CA neurones is enhanced in SHR.F.G. was supported by The Royal Society. SK and JFRP are supported by BHF.1. Souslova EA et al. BMC Biotechnol 7:37, 2007
Rap1 mediates protective effects of iloprost against lung endothelial dysfunction and vascular leak
Authors: Birukov Konstantin G , Fu Panfeng , Xing Junjie , Birukova Anna A 
Published: June 2021

Prostaglandin I2 has been implicated in the regulation of vascular function, inflammation, and acute lung injury. However molecular mechanisms of prostaglandin I2 protective effects on pulmonary endothelial cells (EC) are not well understood. We tested a role of novel cAMP‐activated Epac‐Rap1 pathway in barrier protective effects of synthetic analogue of prostaglandin I2 iloprost using in vivo and in vitro models of lung vascular leak. Iloprost dramatically reduced bronchoalveolar lavage (BAL) protein content, neutrophil accumulation in BAL and lung tissue, capillary filtration coefficient, and Evans blue albumin extravasation caused by high tidal volume ventilation (HTV, 30 ml/kg, 4 hrs). SiRNA‐based Rap1 knockdown inhibited protective effects of iloprost against HTV‐induced lung injury. In vitro, iloprost induced sustained TER increase in lung microvascular endothelium, enhanced intercellular junctions and formation of actin cortical rim, and attenuated thrombin‐induced EC barrier disruption. Consistent with in vivo experiments, Rap1 depletion attenuated protective effects of iloprost in the thrombin model of EC permeability. These data demonstrate that Rap1‐dependent signaling is a major contributor to protective effects of iloprost against lung endothelial barrier dysfunction and vascular leak.HL87823, HL58064
Altered renal hemodynamic and excretory function in rats treated with a COX2 inhibitor during the nephrogenic period
Authors: Reverte Virginia , Saez Fara , Salazar Francisco , Llinas Maria T , Salazar F. Javier 
Published: June 2021

The administration of a COX‐2 inhibitor during the perinatal period (COX2 Inh‐pp) reduces nephron number by 17%, and leads to the development of hypertension. However, there are important gender differences in the aging‐dependent renal structural changes in COX2 Inh‐pp‐treated rats. Our objective was to examine whether the renal response to an amino acid (aas) infusion, and the renal excretory response to an acute volume expansion (AVE) are altered in COX2 Inh‐pp‐treated rats. Newborn SD rats were treated with vehicle or a COX2 Inh‐pp (Rofecoxib, 2.4 mg/kg/day). The responses to aas infusion or to AVE (6%, bw) were examined at 3 months of age in male and female rats treated with vehicle or the COX2 Inh‐pp. No changes in arterial pressure were found in any group during the experiments. An increase (P<0.05) in GFR (47%), RPF (28%) and sodium excretion (152%) was found in vehicle‐treated males in response to aas. However, this renal hemodynamic and excretory response to aas was completely abolished in COX2 Inh‐pp‐treated males. The renal response to aas was also prevented in COX2 Inh‐pp‐treated females. The renal excretory ability to eliminate an AVE was also reduced in male (28%, P<0.05) and female (38%, P<0.05) COX2 Inh‐pp‐treated rats.Our results demonstrate that the COX2 inhibition during the perinatal period leads to an important impairment of the renal hemodynamic and excretory function during the adult age.
Absorption and biosynthesis of D‐chiro‐inositol in mice
Authors: Gopalan Chaya , Lin Xiaobo , Ma Lina , Ostlund Richard 
Published: June 2021

D‐chiro‐inositol (DCI) is known to possess insulin‐like properties. Mice were fed with either regular chow (control group) or DCI‐free diet over a five‐week period. The levels of DCI declined significantly in plasma (1.91 ± 0.17 to 0.056 ± 0.0 μM), stool (20.2 ± 6 to 0.147 ± 0.05 nmoles) and urine (27.39 ± 8.2 to 0.88 ± 0.09 μM) in mice fed with DCI‐free diet at the five‐week period compared to control (plasma 2.47 ± 0.3 to 2.41 ± 0.29 μM; stool 25.7 ± 8.67 to 31.6 ± 9.7 nmoles and urine 39.9 ± 10.3 to 51.5 ± 6.8 µM).Whether DCI could be synthesized endogenously was tested next in the mice maintained on DCI‐free diet for 10 weeks where deuterated water was administered (20 % w/w deuterium oxide) for one week followed by a 24‐hour urine collection. The concentration of deuterium oxide was 8.7 + 0.52 % but there was no natural or deuterated DCI (<0.0067 nmoles/mL) in urine. Instead, there was deuterated myo‐inositol (dMI; 34.4 + 2.8 %) synthesis. To test whether MI could be converted to DCI in mice treated with DCI‐free diet, dMI (1mg MI‐d6) was injected intraperitoneally and urine was collected. Although MI‐d6 constituted 38.7+11.5 % of urinary MI, there was no natural or deuterated DCI present (<0.0067 nmoles/mL). Thus, DCI appears to be derived solely from the diet and may represent additional nutritional factor related to insulin action.
Rapamycin prevents the post‐exercise increase in protein synthesis and alters cell signaling in human skeletal muscle
Authors: Drummond Micah John , Fry Christopher S. , Glynn Erin L. , Dreyer Hans C. , Dhanani Shaheen , Timmerman Kyle L. , Volpi Elena , Rasmussen Blake B. 
Published: June 2021

Resistance exercise stimulates mTOR signaling and muscle protein synthesis (MPS) during post‐exercise recovery in humans. Rapamycin is a specific inhibitor of mTOR and is often used to inhibit protein synthesis in cell models and rodent skeletal muscle in vivo. We hypothesized that the administration of rapamycin to human subjects prior to a bout of resistance exercise would prevent the increase in MPS during early post‐exercise recovery. We studied young men that ingested rapamycin (12mg) 2 hr prior to exercise (Rapamycin; N=7) and a Control group that did not receive rapamycin (N=8). Rapamycin and Control subjects performed a bout of heavy resistance exercise and muscle biopsies were obtained at 1 and 2 hours post‐exercise. Stable isotope and immunoblotting techniques were used to determine MPS and phosphorylation of proteins involved in the regulation of MPS. We found that MPS increased during post‐exercise recovery in the Control group (P<0.05) but was unchanged in the Rapamycin group (P>0.05). In addition, phosphorylation of mTOR (Ser2448), S6K1 (Thr389) and S6 (Ser240/244 and 235/236) were partially inhibited and S6K1 (Thr421/Ser424), eEF2 (Thr56) and ERK1/2 (Thr202/Tyr204) and MNK1 (Thr197/202) were completely blocked during post‐exercise recovery in the Rapamycin group. Raptor association with mTORC1 also tended to be higher in the Control group (as compared to the Rapamycin group) at 2 hours post‐exercise (P=0.06). We conclude that the increase in MPS following an acute bout of resistance exercise appears to be rapamycin dependent indicating that mTOR signaling is playing an important role in regulating the post‐exercise protein anabolic response.Supported by NIH/NIAMS grant # RO1 AR049877 and P30AG024832.
Anti‐inflammatory activities of new steroidal antedrugs isoxazoline derivatives
Authors: Errahali Younes Jean , Gassett Sandi , Lee Henry Joung 
Published: June 2021

Antedrug design represents a new approach projected to create safer drugs by inactivating the metabolites after biotransformation. Our primary objective is to synthesis a new group of corticosteroids that have an anti‐asthmatic and anti‐inflammatory purpose without systemic adverse effects.A new antedrugs isoxazoline derivative is synthesized from prednisolone. Various concentrations of isoxazoline derivatives synthesis was tested by ELISA in vitro: human 549 alveolar epithelium and leukocyte macrophage cells after stimulation with cytomix‐induced proinflammatory molecular release and in vivo: receptor‐binding assay using rat lung and liver cytosol. The metabolism of biotransformation was studied in rat plasma using HPLC technique.Most of isoxazoline derivatives showed high binding affinities to the glucocorticoid receptors; five times more potent than prednisolone and significant inhibitory effects of the antedrugs on the nitric oxide (NO) and other proinflammatory cytokines such IL8 cell release. The metabolism study in rat plasma showed that a new isoxazoline with 21‐acetyl groups were hydrolyzed rapidly to inactive metabolites, with half lives 5 minutes.These results suggest that isoxazolines derivatives compared to conventional steroids improve topical anti‐inflammatory activity without concomitant increase in adverse systemic activity. Support by NIH grants S06 GM08111
The Axial Musculature of the Red Panda (Ailurus fulgens)
Authors: Julik Emily Jean , Smith Heather F. , Adrian Brent , Barton Michael , Fisher Rebecca E. 
Published: June 2021

The red panda (Ailurus fulgens) is an endangered, scansorial carnivore found in the forests of the Himalayas and southern China. The phylogeny of the red panda has been the subject of debate, with molecular and morphological studies indicating a number of possible relationships, including affinities to ursids, procyonids, mustelids, and mephitids. The myology of the red panda, however, remains largely undocumented. The axial muscles of the red panda were evaluated to determine whether any evolutionary affinities with suggested relatives existed, and whether any adaptations to the red panda's scansorial lifestyle were present in these muscles. Two adult red pandas were dissected and photodocumented. The axial musculature of Ailurus was then compared to other carnivores. The dissections revealed several interesting findings. For example, m. biventer cervicis was found to have six tendinous intersections, similar to those found in the mm. rectus abdominis and likely allowing precise movement of the neck just as the rectus muscles allow for precise movements of the trunk. The comparison of the axial musculature of the red panda to that of other carnivores also revealed interesting findings; for example, the red panda had several differences from the giant panda (Ailuropoda melanoleuca), including a more superficial m. splenius with a significantly more cranial origin.Support from the Howard Hughes Medical Institute through the Undergraduate Science Education Program and from the ASU School of Life Sciences.
SNAP‐tag, A Multipurpose Visualization Tool for Studying Protein Dynamics in Living Cells
Authors: Sun Luo , Zhang Aihua , Ghosh Inca , Xu Ming‐Qun 
Published: June 2021

The creation of cell lines expressing proteins fused to intrinsically fluorescent tags (e.g., GFP) and their use in cell based studies of protein localization have become routine. The fluorescent signal from such tags cannot be routinely switched on at will, precluding some time‐resolved studies of expression, localization and degradation.A complementary and more flexible imaging system based on a family of small, self‐labeling protein tags is being introduced by New England Biolabs. SNAP‐tag and CLIP‐tag, orthogonal tags with a variety of fluorescent substrates, are suitable for imaging intracellular and cell surface proteins in live cells. These tags can also be used for time‐resolved dual labeling, highly efficient covalent pull‐downs and other biochemical assays. Furthermore, a collection of non‐fluorescent substrates that block SNAP‐ and CLIP‐tag reactivity enable pulse‐chase studies and an examination of the temporal dynamics of nascent protein synthesis and complex formation in live cells.In this report, several studies of protein localization, turnover and receptor internalization using SNAP‐ and CLIP‐tags in living cells are demonstrated. Using two fluorescent dyes in pulse‐chase experiments, we examined the turnover of several intracellular proteins including the Cytochrome c oxidase subunit 8A (COX8A) and the Transferrin Receptor. We demonstrate that fluorescent protein labeling using SNAP‐tag and CLIP‐tag provides a powerful tool to study protein localization, turnover and internalization.
Non‐canonical Wnt signaling regulates the stability of xSyndecan4
Authors: Escobedo Noelia A. , Carvallo Loreto , Munoz Rosana , Larrain Juan 
Published: June 2021

The non‐canonical or beta‐catenin independent Wnt signaling pathway regulates convergence and extension movements in vertebrate embryos and planar cell polarity. Recently in our laboratory, we have demonstrated that a cell surface heparan sulphate proteoglycan, Syndecan4 (xSyn4), is critical for Xenopus laevis gastrulation and is a component of the non‐canonical Wnt signalling pathway. We have found that xSyn4 is expressed in the same tissues and interacts functionally and biochemically with Frizzled‐7 (xFz7) and Dishevelled (xDsh). Furthermore, xSyn4 is necessary and sufficient to activate translocation of xDsh to the plasma membrane. In this work we have used methodologies such as GST‐pulldown and immunoprecipitation in HEK293T cells in order to investigate the interaction between xSyn4, xDsh and xFz7. We have observed that xDsh and xFz7 interact with xSyn4 cytoplasmic domain. Also, we have found that xSyn4 stability is regulated by activation of the non‐canonical Wnt pathway and that the cytoplasmic domain is required for such effect.This work is supported by FONDECYT Post‐Doctoral Fellowship 3070015 to L.C. and the Center for Cell Regulation and Pathology (FONDAP).
Role of S100A8 and S100A9 proteins on breast cancer aggressivity
Authors: Cormier Kevin A , Fournier Sebastien , O'Brien Jeannick , Tessier Philippe , Robichaud Gilles A 
Published: June 2021

Women are more affected by breast cancers than any other type of cancer. Studies show that proteins from the S100 family are differentially expressed in cancers and autoimmune disease. The S100 protein family is the largest family of calcium binding proteins where most members are overexpressed in certain types of cancers. More specifically, S100A8 and S100A9 seem to be overexpressed in mammary ductal carcinomas. Nevertheless, little is known on the cellular effects of S100 proteins and their roles in breast cancer pathogenesis.In this study, we assessed cancerous biological processes (proliferation, apoptosis, cell adherence and ECM invasion) in mammary cells following: 1) the treatment with various concentrations of recombinant and purified S100A8 and S100A9 proteins and, 2) the knocked‐up or knocked‐down expression of the S100A8 and S100A9 genes. We found that extracellular treatments with S100 proteins affected growth rates of cancer cells. We also observed that the conditioned expression of S100 protein in breast cancer cells affected cancer processes in a cell type dependent manner. Details from these results will be discussed here.These results will aid to a better understand the roles of S100A8 and S100A9 proteins in epithelial cell biology and breast cancer pathogenesis. The outcome of these experiments could potentially lead to a better diagnostics or more specialized treatments of breast cancers.
A Targeted Therapeutic Rescues Botulinum‐A Poisoned Neurons
Authors: Zhang Peng , Ray Radharaman , Singh Bal Ram , Li Dan , Adler Michael , Ray Prabhati 
Published: June 2021

Botulinum neurotoxin (BoNT) is the most poisonous poison known to mankind for which currently no antidote is available to rescue poisoned nerves. An effective medical countermeasure strategy will be required and would include its efficient delivery specifically to presynaptic nerve terminals. Here we present the report of rescue of botulinum poisoned nerve cells by Mastoparan‐7, a peptide constituent of bee venom that was delivered through a drug delivery vehicle (DDV) constructed from the non‐toxic fragment of botulinum neurotoxin itself. We found that mastoparan‐7 that was delivered into spinal cord cells rescued BoNT/A poisoned cells by restoring over 40% neurotransmitter release. The rescue of the cell poisoning does not seem to occur from inhibition of the endopeptidase activity of BoNT/A against its well known substrate, SNAP‐25, mechanistically involved in the exocytosis process. Rather, Mastoparan‐7 induces a physiological host response apparently unrelated to SNAP‐25 but linked to the phospholipase signal transduction pathway. Our results open new avenues to examine mechanism of exocytosis process, and also to examine alternative cellular mechanism of botulinum neurotoxin action. Successful demonstration of BoNT‐based DDV opens door for its utility for drug delivery against many neuronal and neuron‐muscular disorders.This work was supported JSTO‐CBD/DTRA #3.10003_05_WR_B
Therapeutic Effect of Ginkgo Biloba Extract on Rat Heart Failure Induced by Isoprenaline
Authors: Yi Hongwei , Wang Yongqing , Ding Fuyun , Meng Ling , Wu Xudong 
Published: June 2021

ObjectiveTo investigate the effect of Ginkgo biloba extract (GBE) on rat heart failure induced by isoprenaline and its mechanism.MethodsThe SD rat heart failure was induced by isoprenaline administrated subcutaneouly for 11 days. 100 mg/kg/d GBE and 10 mg/kg/d Metoprolol were pretreated 3 days before isoprenaline. The index of haemodynamics and serum contents were detected.ResultsIsoprenaline increased the heart index and left ventricle index significantly, but GBE and Metoprolol could alleviate rat myocardial hypertrophy. GBE had stronger effect on inhibition of the increase in AST and LDH than Metoprolol; on the contrary, Metoprolol was more potential for decreasing creatine kinase, MB isoenzyme of creatine kinase and hydroxybutyrate dehydrogenase level. Isoprenaline decreased the level of LVSP, LVEDP, LV +dp/dt max and LV ‐dp/dt max and increased heart rate. Metoprolol and GBE blocked the effects of isoprenaline respectively, but there is not significant difference between GBE and Metoprolol. GBE and Metoprolol inhibited the increase in mRNA levels of AMP‐activated protein kinase alpha 1, 2 and ryanodine receptor 1 induced by isoprenaline significantly.ConclusionGBE attenuated the damage of cardiac muscle, improved the dysfunction of rat heart induced by isoprenaline. Not like Metoprolol, a beta receptor blocker, GBE exerts a different mechanism in curing heart failure.
Reduction of the trans fatty acid absorption in Caco‐2 cell by grain extracts.
Authors: Lee Sung‐Joon , Jeun Jungae , Cho Hee‐Jun , Shin Ki‐Yong 
Published: June 2021

Reduction of the trans fatty acid absorption in Caco‐2 cell by grain extracts. Jungae Jeun, Hee‐Jun Cho, Ki‐Yong Shin, Sung‐Joon LeeIntake of trans‐fatty acids could increase LDL‐cholesterol and reduce HDL‐cholesterol levels thus may increase the risk for coronary diseases. We investigated the inhibition of grain extracts on the elaidic acid (t18:1) uptake, predominant trans‐fatty acid in the processed foods. Water, ethyl acetate, and hexane extracts from white and black soybeans, barley, brown rice, and wheat powder were used in the study. The composition profile of each extract was analysed by GC‐MS analysis and the effects of each extracts on the elaidic acid absorption on Caco‐2 cells were measured by GC analysis. RT‐PCR and western blot were performed to assess gene expression in the cellular fatty acid uptake. Hexane extracts of black soybean was the most effective and inhibited 15% of elaidic acid uptake, and down regulated the expression of CD36 and FATP2 by 54% and 49%, respectively. ACS1, FABP, NPC1L1 were marginally altered. These suggest that soybean may reduce trans‐fatty acid uptake by altering the expression of fatty acid transporters in the small intestine.
Osteologic exploration of the clavicle: a new approach
Authors: Bortier Hilde Elisa , Bernat Amit , Huysmans Toon , Van Glabbeek Francis , sijbers jan , Pinho Romulo , Gielen Jan , Hubens Guy 
Published: June 2021

IntroductionClavicles have a complex osteologic structure which makes a morphometric analysis extremely difficult. Our analysis shows the exact measurements and variations of the clavicle.Materials and Methods90 clavicles were dissected, CAT scanned and reconstructed. All measurements were automatically performed. The length and curvatures were calculated around the central line and for each cross‐section the average, sagittal and axial diameter was calculated.ResultsThe average length is 163±11 mm. For the length, there is a 9% difference between the gender and 1.2% between the left and right clavicle. Between the genders there is a volume difference of 36%. The extremities show the biggest diameter, this decrease as approaching to the inflexion point, which is the smallest average diameter. In the axial view the acromial curvature is shorter and more curved than the sternal one. In males, the maximum acromial curvature has a difference of 18% and the maximum sternal curvature a difference of 4% compared with the females. In the coronal view there is a concave curvature with a maximum of 6 mm. In females the acromial end bends more posteroinferiorly.DiscussionThis is the first 3D analysis performed on the clavicle. Females have a smaller clavicle with a shorter and less curved acromial curvature and a posteroinferior bending. The right clavicle is slightly shorter, thicker and more robust.