bluuebunny/crossref_metadata_2025_split huggingface dataset:
Impact of concomitant use of opioids and benzodiazepines in opioid‐dependent pregnant women on the outcome of their newborns
Authors: Metz Verena , Comer Sandra D. , Fischer Gabriele 
Published: June 2021

OBJECTIVEThe study objective was to compare neonatal outcome parameters of children born to opioid‐dependent women in opioid maintenance therapy with differing concomitant consumption patterns during the last trimester of pregnancy.METHODSAn observational design and t‐tests were applied to analyze the outcome of 106 neonates born to opioid‐maintained, smoking mothers with (n=27) versus without (n=79) concomitant consumption of opiates and benzodiazepines.RESULTSNewborns of mothers with concomitant opiate and benzodiazepine consumption had significantly higher average withdrawal scores (8.27 vs. 6.15, p=0.007) and significantly longer postnatal hospital stays (26.5 vs. 18.6 days, p=0.011) compared to those of mothers without problematic concomitant consumption. A subgroup analysis confirmed the results for the 62 newborns whose mothers were maintained on methadone throughout pregnancy, but these differences could not be shown in the 44 children of buprenorphine‐maintained mothers.CONCLUSIONSThese findings suggest that the opioid maintenance medication might have a stronger influence on neonatal outcome parameters than concomitant consumption of other substances during the last trimester.
A new molecular sensor controlling cAMP activation of ERK
Authors: Emery Andrew Clayton , Eiden Maribeth V , Mustafa Tomris , Eiden Lee E 
Published: June 2021

The signal transduction pathway linking cAMP elevation through GPCR‐Gs stimulation to ERK activation is incompletely understood. We recently reported that cAMP‐ and ERK‐dependent neuritogenesis in NS‐1 cells initiated by activation of the GPCR PAC1 occurs independently of both PKA and Epac activation, revealing the existence of an additional cAMP sensor (NCS, for neuritogenic cAMP sensor) (Emery and Eiden, FASEB J, 26: 3199, 2012). The adenylate cyclase (AC) inhibitor SQ22,536 blocks cAMP‐dependent ERK activation without affecting either CREB phosphoryation or Epac‐dependent Rap1 GTP loading, thus identifying NCS as a new target of SQ22,356 (see Emery et al., doi:10.1124/mol.112.081760). The pharmacological profile of the NCS has facilitated its molecular identification as a member of the Ras guanine nucleotide exchange factor superfamily. The characteristics of NCS cAMP binding studied in bovine chromaffin cells, and NCS‐dependent ERK signaling studied via gain‐of‐function in HEK293 cells, suggests a previously unremarked and neuroendocrine cell‐specific signaling pathway, GPCR→Gs→AC→cAMP→NCS→Rap1→Braf→MEK, connecting neuropeptide receptor occupancy to ERK activation.Supported by NIMH‐IRP project ZO1‐MH002386.
Functional interactions of a monothiol glutaredoxin and an iron sulfur cluster carrier protein with the radical SAM enzyme MiaB
Authors: Butland Gareth , Boutigny Sylvain , Saini Avneesh , Baidoo Edward E.K. , Yeung Natasha , Keasling Jay D. 
Published: June 2021

The biosynthesis of iron sulfur (FeS) clusters is a highly coordinated process enabled by multi‐protein systems such as Isc and Suf in E. coli. While these systems are believed to encode all factors required for initial cluster assembly and transfer to FeS carrier proteins, accessory factors of undefined function, including the monothiol glutaredoxin, GrxD and the FeS carrier protein NfuA, are located outside of these defined systems. These factors have been suggested to function both as shuttle proteins acting to transfer clusters between scaffold and carrier proteins, and in the final stages of FeS protein assembly by transferring clusters to client FeS apo‐proteins. Here we implicate both of these factors in client protein interactions. We demonstrate specific interactions between GrxD, NfuA and the methylthiolase MiaB, a radical SAM enzyme involved in tRNA maturation. We show that GrxD and NfuA physically interact with MiaB with affinities compatible with an in vivo function. We furthermore demonstrate that NfuA is able to transfer its cluster in vitro to MiaB, while GrxD is unable to do so. The relevance of these interactions was demonstrated by linking the activity of MiaB with GrxD and NfuA in vivo. We observe a severe defect in in vivo MiaB activity in cells lacking both GrxD and NfuA, suggesting that these proteins may play complementary roles in the MiaB catalytic cycle.Supported by NIH award GM088196 to GB.
The effect of EMD 87580, an Na+/H+ isoform 1 inhibitor, on the rate of cell proliferation in mouse lung adenocarcinoma cells
Authors: Mraiche Fatima , Alasmar Aya Riyad , Al Marridi Wafa Ziad , Youssef Maiy 
Published: June 2021

BackgroundDysregulated pH is an emerging hallmark that maybe permissive for cancer progression. The Na+/H+ exchanger (NHE1) is one of the most notable pH regulating transporters and as such warrants further investigation for its role in non small cell lung cancer (NSCLC).MethodsNHE1 protein expression and the rate of cell proliferation were examined in cultured mouse lung adenocarcinoma cells ±10μM NHE1 inhibitor (EMD 87580) or Etoposide for 24 or 48 hours using western blotting and the MTT assay, respectively.ResultsWestern blotting demonstrated an increase in NHE1 protein expression in mouse lung adenocarcinoma cells. In the presence of EMD 87580 (N), NHE1 protein expression showed a trend towards reduction vs. control group (C) (100% C vs. 61.5% N). Treatment of mouse lung adenocarcinoma cells with EMD 87580 for 24 hours did not alter the rate of cell proliferation. Similarly, Etoposide, an anticancer topoisomerase inhibitor, had no effect on the rate of cell proliferation following 24 hours of treatment (92.8±6.3%). However, 48 hours of treatment with EMD 87580 revealed a trend towards reduction in the rate of cell proliferation in mouse lung adenocarcinoma cells (100±11.3% C vs. 77.4±2.6% N). The rate of cell proliferation was significantly less in mouse lung adenocarcinoma cells treated with Etoposide (34.9±7.3%).ConclusionTaken together, these data demonstrate that NHE1 protein was over expressed in the mouse lung adenocarcinoma cells. NHE1 inhibition regresses cell proliferation with prolonged treatment. It is evident the tumor progression and metastasis require NHE1.
TRPA1 channels play a critical role in cold‐induced vasodilatation
Authors: Graepel Rabea , Aubdool Aisah A , Kodji Xenia , Fernandes Elizabeth Soares , Bevan Stuart , Brain Susan D 
Published: June 2021

The mechanisms involved in cold‐induced vasodilatation are unclear. Here, we have investigated an involvement of the cold‐sensitive channel TRPA1, known to be expressed in a subset of sensory neurons. Using laser Doppler flowmetry, peripheral blood flow was measured following cold exposure in the plantar skin in the anaesthetised mice before (5 min:baseline) and after local cold exposure of the hindpaw (30 min). This response consisted of a decrease in blood flow, followed by an increase in WT mice. This response was substantially reduced in TRPA1 KO as compared to WT mice (245.1 ± 39.6 vs 86.1 ± 11.0, 103 flux units, measured as area under the response curve, n=6, p<0.01) and antagonised by the TRPA1 antagonist HC030031 (100mg/kg, i.p., n=5). The cold‐induced response was also shown to be significantly reduced in WT mice pre‐treated with the calcitonin gene related peptide (CGRP) receptor antagonist, CGRP8–37 and the neurokinin‐1 receptor antagonist SR140333, suggesting a prominent role of neuropeptides in this response. These results suggest the major involvement of TRPA1 containing sensory nerves in local cold‐induced vascular responses.Supported by a BBSRC‐led IMB capacity building award.
Sildenafil restores endothelial function and reduces blood pressure in experimental hypertension
Authors: de Moura Balarini Camille , Gomes Isabele Beserra Santos , Dias Ananda Tissianel , Fahning Bernah Mathias , Cintra Amanda , Frossard Jessica , Gava Agata Lages , Meyrelles Silvana dos Santos , Vasquez Elisardo Corral 
Published: June 2021

The aim of this study was to evaluate the effects of sildenafil on endothelial function of hypertensive mice. Male C57BL/6 mice (n=3–10) were subjected to renal artery clipping (2K1C). After 14 days, sildenafil (40 mg/kg/day) or vehicle was administrated for 14 days. At the end of the treatment, mice were anesthetized and catheterized to blood pressure recording and had the mesenteric arteriolar bed (MAB) and thoracic aorta (TA) removed to vascular function studies. Data are mean ± SEM. One‐way ANOVA was performed followed by Tukey's post hoc test. Sildenafil caused a significant reduction in the arterial blood pressure in treated mice (126±3 vs. 110±4mmHg). Endothelial dysfunction was observed in MAB of 2K1C mice, but not in the TA and it was restored by sildenafil (Rmax ACh: 44±2.6% vs. 59±4.5%). Ongoing studies are revealing an augmented participation of nitric oxide in this process in treated animals. Similarly, an increased maximal response to an alpha 1‐agonist was observed in MAB but not in TA in 2K1C mice, which was ameliorated by sildenafil (167±8 vs.117±9 mmHg). Both TA and MAB did not show impairment in smooth vascular relaxation to sodium nitroprusside. Thus, our data showed that sildenafil can restore endothelial function in hypertensive mice, which reflects in a significant reduction of arterial hypertension.Financial Support: CNPq (Proc. 302582/2011–8) and FAPES/Universal 012/2011 (Proc. 54498465).
Circulating plasma factors contribute to age‐associated arterial stiffness
Authors: Johnson Lawrence Cody , Eng Jason S. , Fleenor Bradley S. , Seals Douglas R. , Sindler Amy L. 
Published: June 2021

Previous findings from our laboratory showed that age‐related increases in large elastic artery stiffness in mice are associated with remodeling of the extracellular matrix (ECM), as indicated by increased collagen I expression, and oxidative stress, as indicated by increased nitrotyrosine staining. Here we tested the hypothesis that circulating factors in plasma contribute to arterial oxidative stress, ECM remodeling and stiffening. Nitrotyrosine, collagen I and elastic modulus, a measure of intrinsic mechanical stiffness, were determined in aortic rings from young (6 mo, n=4) male C57 mice cultured for 72 hours in media containing 10% plasma from either young (Yplasma) or old (Oplasma) mice. Compared with Yplasma, aortic rings incubated in Oplasma demonstrated increased NT abundance (3.11 ± 0.50 vs. 1.00 ± 0.30 AU, p<0.05), collagen I (1.66 ± 0.18 vs. 1.00 ± 0.12 AU, p<0.05) and elastic modulus (2780 ± 570 vs. 1768 ± 188 kPa, p=0.05). These preliminary results are consistent with the hypothesis that unknown factors in the circulating plasma induce “aging‐like” oxidative stress, ECM remodeling featuring increases in collagen I, and stiffening of the aorta. Identifying these circulating factors may lead to additional therapeutic targets for the prevention and treatment of ageassociated large elastic artery stiffening and clinical cardiovascular diseases.Supported by NIH AG013038, AG000279, HL007822
Shortened life span in mice generated by in vitro fertilization
Authors: Rexhaj Emrush , Rimoldi Stefano , Pireva Agim , Robert Von Arx , Allemann Yves , Sartori Claudio , Scherrer Urs 
Published: June 2021

In humans assisted reproductive technologies (ART) induce generalized vascular dysfunction in children. In line with this observation, ART induces vascular dysfunction and arterial hypertension in mice that is related to an epigenetic mechanism, but the long term consequences are unknown. To address this problem, we compared the long term survival of ART and control mice fed a normal chow diet (NC) or challenged with a high‐fat diet (HFD).80 male FVB mice generated by ART and 80 control mice born and raised in our animal facility were observed in these studies. Three to four animals were placed in a cage in the clean conventional mouse facility. 6 ART and 7 control mice had to be sacrificed because of injuries related to fighting. 32 ART and 20 control mice were fed NC, 42 ART mice and 53 control mice were fed a HFD (49.5% fat, 31.5% protein and 0% carbohydrate). Kaplan‐Meier survival curves were constructed using known birth and death dates and the log‐rank test was used to evaluate statistical differences between groups.We found that in mice fed with NC the life span was similar in the two groups. In contrast, HFD, while not having a detectable effect on life span in control mice, reduced the survival in ART‐mice by roughly 25%(P<0.0001).ART is associated with a marked reduction of the life span in normal mice challenged with a Western style high‐fat diet.
Study of the roles of LPA3 on erythropoiesis and thrombocytopoiesis processes in zebrafish
Authors: LI MENG WEI , Huang Chang‐Jen , Lee Hsinyu 
Published: June 2021

Study of the roles of LPA3 on erythropoiesis and thrombocytopoiesis processes in zebrafishHematopoiesis is a dynamic process regulated by complicated pathways. It is responsible for the production of hematopoietic stem cells, as well as their differentiation into more specific mature cell types. Lysophosphatidic acid (LPA), an extracellular lipid mediator, binds to its cognate G protein‐coupled receptors, through which can regulates differentiation and development of the circulation system. In previous study, we demonstrated that LPA regulates hematopoiesis determination in stem cells and erythropoiesis processes through activating LPA receptor 3 (LPA3). Since erythrocytes and thrombocytes were derived from common progenitors, we aim to investigate whether LPA3 also regulates thrombocyte development. Using zebrafish as a model, we found that morpholino (MO)‐mediated knockdown of LPA3 inhibited production of hemoglobin using o'dianisidine staining of zebrafish embryos. Furthermore, treatment with OMPT, a LPA3 agonist, and Ki16425, a LPA1 and LPA3 antagonists, were used to determine if the differentiation process of erythrocyte or thrombocyte is affected. Our results show that treatment with Ki16425 leads to erythropoiesis defects. In addition the expression levels of CD41 (αIIb, glycoprotein IIb [GPIIb]), a marker for thrombocyte, was increased by LPA3 MO and decreased by with OMPT treatments. In summary, our results demonstrated that LPA3 is an important regulator for erythropoiesis and thrombocytopoiesis process of hematopoietic stem cells.
NMR Studies of the DNA Polymerase β L22P Variant
Authors: KIRBY THOMAS 
Published: June 2021

The gastric cancer‐associated DNA Polymerase β (Polβ) variant with Lys22 replaced by Pro (L22P), has been shown to exhibit reduced DNA binding as well as reduced DNA lyase activity, and to be deficient in base excision repair [Dalal,S., Chikova,A., Jaeger,J., and Sweasy,J.B. (2008) Nucleic Acids Res., 36, 411–422]. The amino acid change is in the middle of helix1, the first of 4 helices that make up the Polβ lyase domain.We have used NMR experiments with labeled L22P to probe the structure of the variant in solution. The 1H‐13C HSQC resonances of 13C methyl methionine‐labeled enzyme indicate that the L22P mutation appears to structurally disrupt at least part of the lyase domain, as judged by the chemical shift and intensity of the M18 resonance. The rest of the methionine methyl resonances, which arise from the polymerase domain, are unaffected by the mutation. The methionine methyl resonances of wild‐type Polβ exhibit characteristic perturbations upon formation of binary and ternary complexes with gapped DNA and a complementary dNTP. Under the same conditions, the methionine resonances of L22P show none of the changes associated with complex formation. Consistent with these observations, the 1H‐15N HSQC spectrum of the isolated 8K lyase domain of the L22P Polβ variant is characteristic of a disordered protein.
Higher intakes of low‐fat milk combined with 12 weeks of endurance training does not result in lower fat mass and higher lean mass.
Authors: Roberts Rebekah , Euiler Elizabeth , Ward Wendy , LeBlanc Paul , Tarnopolsky Mark , Roy Brian 
Published: June 2021

Previous studies showed that a low‐fat milk intervention combined with resistance training resulted in lower fat mass and higher lean mass in males and females. The purpose of this study was to determine the effects of combining increased low‐fat milk intake with endurance exercise on fat and lean mass. 32 healthy young males were randomized and blindly assigned into one two groups 1) MILK (3 additional servings of low‐fat milk, 750 mL) or 2) CHO (maltodextrin beverage, isocaloric to MILK). All participants completed a 12‐week endurance‐training program. The exercise program consisted of cycling 1 hour per day at ~60% VO2 peak, 5 days per week for 12 weeks. Whole body fat and lean mass were determined using dual‐energy X‐ray absorptiometry (DXA) that was performed before and after training. 23 participants completed the study. Changes in body mass (MILK: 0.39±0.61 kg; CHO: 0.30±0.77 kg), fat mass (MILK: −420.7±522.1 g; CHO: 99.6±794.8 g) and lean mass (MILK: 831.2±509.8 g; CHO: 624.9±434.6 g) were similar between the 2 groups. Findings to date suggest that combining a higher intake of low‐fat milk with 12 weeks of endurance training does not improve changes in body composition. Further analyses are ongoing.Funding provided by The Dairy Farmers of Canada.
Excitatory effects of neurokinin 2 and bombesin receptor peptide agonists vivo in urinary tract of rats with voiding dysfunctions
Authors: Kullmann Florenta Aura , Zheng Jihong , Wells Grace , McKenna David , Burgard Ed , Thor Karl 
Published: June 2021

Bladder function was examined in anesthetized and awake adult female Sprague Dawley rats. Voiding dysfunction models included STZ‐induced diabetic (streptozotocin; STZ; 65mg/kg i.p., 9–20 weeks post treatment), polyuria (5% sucrose‐fed; >;3 weeks post treatment) and spinal cord injured (SCI; T8‐T9 cut; acute or 3 days – 6 weeks post SCI) rats. Telemetry transmitters were implanted into polyuria group to measure bladder pressure in freely moving rats. [Lys5,MeLeu9,Nle10]‐NKA(4–10) (LMN‐NKA), a neurokinin 2 receptor (NK2R) agonist, gastrin‐releasing peptide (GRP), a bombesin receptor 2 (BBR2) agonist, and neuromedin B (NMB), a BBR1 agonist, were delivered via jugular vein catheters.In urethrane anesthetized STZ and control rats, LMN‐NKA (0.1–100μg/kg), GRP (0.1–300μg/kg) and NMB (0.1–300μg/kg) dose dependently increased bladder contraction (BC, up to 4x) and micturition frequency. In STZ rats, LMN‐NKA reduced residual volume by half. In acute anesthetized SCI rats, LMN‐NKA (>;10μg/kg) induced urine release (voiding efficiency ~40%). In awake polyuria rats, LMN‐NKA and GRP (1–100μg/kg) induced BC that was often associated with voiding. In awake SCI rats, LMN‐NKA (100μg/kg) and GRP (100, 300μg/kg) triggered voiding in 25 – 80% of rats.Activation of NK2R and BBRs enhanced or triggered voiding in naïve rats and in several models of voiding dysfunction including STZ and SCI rats.
Induction of Apoptosis by ER Stress in Lung Alveolar Epithelial Cells (AECs) Is Blocked by Angiotensin 1–7 Through Mas
Authors: Uhal Bruce , Nguyen Hang , Dang MyTrang , Gopallawi Indiwari , Dang Vinh , Ono Shinji , Morimoto Konosuke 
Published: June 2021

STUDY OBJECTIVEOur prior work showed that apoptosis of AECs is regulated by autocrine generation of angiotensin (ANG) II and the counterregulatory ANG1–7. Mutations in surfactant protein C (SP‐C) induce endoplasmic reticulum (ER) stress and apoptosis. This study tested the hypothesis that ER stress‐induced apoptosis of AECs might also be regulated by autocrine ANGII/ANG1–7.METHODSA549 cells or primary cultures of human AECs were challenged with MG132 or the SP‐C BRICHOS domain mutant G100S. The ANGII‐generating and –degrading enzymes cathepsin D and ACE‐2 were measured by enzyme assay and western blot. Apoptosis was measured by mitochondrial function, JNK and caspase activation and nuclear fragmentation.RESULTSER stress activated cathepsin D and decreased ACE‐2. Both changes were inhibited by TAPI‐2, a blocker of ADAM17/TACE. Apoptosis of A549 or primary human AECs induced by MG132 or the SP‐C BRICHOS mutant G100S was significantly inhibited by the nonselective ANG receptor blocker saralasin and was completely abrogated by ANG1–7 (both p<0.01). Inhibition by ANG1–7 was blocked by the specific mas antagonist A779.CONCLUSIONSThese data show that ER stress‐induced apoptosis is mediated by the autocrine ANGII/ANG1–7 system in human AECs and demonstrate effective blockade of SP‐C mutation‐induced apoptosis by angiotensin1–7 through a mas‐dependent mechanism.Support: PHS HL‐45136
Tet1/3 mediated epigenetic regulations in male germ cell development
Authors: Luk Alfred CS , Ng Shuk Hang , Tao Jiajie , Chan Wai‐Yee , Lee Tin‐Lap 
Published: June 2021

Previous epigenetic studies focused mainly on 5‐methylcytosine (5mC), emerging evidence suggests that 5‐hydroxymethylated cytosine (5hmc) could be another functional epigenetic modification in eukaryotic genome as demonstrated during differentiation of neurons and embryonic stem cells. However, its roles in male germ cell development remain unexplored.Our preliminary data demonstrated decreasing expression of ten‐eleven translocation (Tet) protein family after serial passages of C18–4 cells (mouse type A spermatogonia), suggesting potential correlation on epigenetics and aging. Moreover, we showed a higher expression level of Tet1/3 in stem‐cell like C18–4 cells than in somatic TM4 (mouse Sertoli) cell. Our dot‐blot results also demonstrated a higher genomic 5hmC content in C18–4 than in TM4 cells, implying possible stemness‐related regulation of 5hmC. Taken together, these data revealed a potential regulation of 5hmC/Tet in male germ cell development, which furthered our knowledge on male reproductive health and infertility control.
Role of endothelial sialic acid expression on xenogenic neutrophil adhesion (1003.4)
Authors: French Beth , Harris Donald , Benipal Prabh , Pierson Robin , Azimzadeh Agnes 
Published: June 2021

Objective: Transplantation of organs across species, xenotransplantation, is limited by the sequestration of platelets and neutrophils in injured organs; we hypothesized that sialic acids on porcine endothelial cells affects adhesion of human neutrophils.Methods: An adhesion assay was developed with freshly isolated human neutrophils and porcine aortic endothelial cells (pAECs). Neutrophils labeled with 0.5 uM of Calcein were quantified using a Spectramax fluorescent reader. Neutrophil adhesion was expressed as arbitrary fluorescence units. 25 mU of Neuraminidase (Clostridium Perfringens) was added to remove sialic acids from pAECs 30 minutes before neutrophil addition. Sialic acid expression was assessed by flow cytometry using lectins (Elderberry lectin, (SNA), Peanut agglutinin (PNA) and Maackia amurensis hemoagglutinin (MAC)). Some pAECs wells were incubated with human TNF (25ng/ml) for 3 hrs at 37 °C before addition of neuraminidase and neutrophils.Results: Human neutrophil adhesion was detectable on resting porcine endothelial cells (52 +/‐15) and was increased by TNF activation (72 +/‐ 4.7). Neuraminidase treatment increased neutrophil adhesion on resting pAECs to a level similar to TNF activation (71 +/‐ 8.8).Conclusion: Removal of sialic acid with neuraminidase increased the adhesion of human neutrophils to swine endothelial cells. Thus, targeting levels of sialic acid expression may be a novel therapy to limit neutrophil adhesion during xenotransplantation. Future studies will examine the role of endothelial sialic acid on platelet adhesion as well as the role of neutrophils and platelet sialic acid expression.Grant Funding Source: Supported by: NIH 5T32HL007698‐19 & U19AI090959; United Therapeutics unrestricted gift