bluuebunny/crossref_metadata_2025_split huggingface dataset:
Association Between Anatomical Measurements and Dynamic Knee Valgus in Healthy Women And Women With ACL Reconstruction
Authors: Rosario Martin G , Ortiz Alexis 
Published: June 2021

Anthropometric measurements have been associated with anterior cruciate ligament (ACL) injury in women.PURPOSEAssess the association between anthropometric measures of the lower extremities and dynamic knee valgus in women with and without ACL reconstruction.METHODSTwenty‐nine women (non‐injured=15, ACL=14) participated in this investigation. Anthropometric measures included: height, weight, inter‐ASIS distance, ASIS‐to‐trochanter distance, leg length, knee width, and ankle width. Dynamic knee valgus was measured with a three‐dimensional motion capture system (120Hz) as the distance between markers placed at the lateral femoral epicondyles during the landing phase of a double‐legged 60‐cm height drop jump. T‐tests were used to compare anthropometric measures between groups. Multiple linear regressions were used to examine the association between anthropometric variables and valgus.RESULTSAll measurements except knee width were statistically significant different (p>.05) between groups with the ACL group exhibiting greater values. Regression analyses showed no statistically significant association for the control group (r2=.51; p>.05), whilst the association for the ACL group was statistically significant (r2=.89; p=.007).CONCLUSIONSIt appears that anthropometric measures are predisposing factors to dynamic knee valgus that in turn are directly related to ACL injury.Grant Funding Source: NIH‐Clinical Research Education and Career Development in Minority Institution R25RR017589
The Anabolic Role of Cannabinoid Receptor in Bone
Authors: Del Carpio‐Cano Fabiola Elizabeth , Marcu Jahan , Popoff Christopher , Pixley Robin , Abood Mary , Popoff Steven , Safadi Fayez 
Published: June 2021

Endocannabinoids and their G protein‐coupled cannabinoid receptors 1 (CB1) and 2 (CB2) are present in the skeleton. CB2 is expressed in osteoblast (OB), though CB1 is debatable. We have begun a characterization of the skeletal phenotype in CB1/CB2 double receptor knockout (CB1/2 dKO) mice, We also utilized specific CB1&2 receptor antagonists to examine their effects, either individually or in combination, on OB differentiation. We conducted micro‐CT analysis of femurs from 12 week‐old CB1/2 dKO mice demonstrating a significant reduction in cortical and trabecular bone volume and thickness, when compared to wild‐type mice. To examine the functional significance of CB1&2 receptors on OB differentiation, we established primary OB cultures utilizing newborn wild‐type calvaria. Alkaline phosphatase (ALP) activity was assessed to evaluate the effects of CB receptor antagonists on OB differentiation. Treatment with the CB1 or CB2 antagonist caused a significant, dose‐dependent inhibition of ALP activity. In cultures treated with both antagonists, the inhibition of ALP was more pronounced compared to either antagonist alone. Our results suggest that CB1 & CB2 are functional receptors during OB differentiation and in regulation of bone mass. Additional studies are warranted to evaluate intracellular signaling subsequent to ligand activation of CB1&2 in OBGrant Funding Source: NIH‐NIAMS AR048892
Maternal expression of atrial natriuretic peptide and the fetal programming of cardiac hypertrophy
Authors: Armstrong David WJ , Tse M Yat , Wong Philip G , Pang Stephen C. 
Published: June 2021

IntroductionThe natriuretic peptide system (NPS) is a key regulator of cardiovascular homeostasis. Mice lacking atrial natriuretic peptide (ANP) display chronic hypertension with salt‐sensitivity and cardiac hypertrophy (CH). The effect of maternal ANP expression on the development of CH in offspring is unknown.ObjectiveTo determine the effect of maternal ANP genotype on the development of CH in ANP heterozygous (ANP+/−) offspring born to either ANP wild‐type (+/+) mothers (ANP+/−WT offspring) or ANP knockout (−/−) mothers (ANP+/−KO offspring).Methods9‐week old male ANP+/−WT and ANP+/−KO mice were treated with normal salt (NS; 0.8% NaCl) or high salt (HS; 8.0% NaCl) diet for 5 weeks. Mice were euthanized, organs were weighed, and left ventricular (LV) gene expression was measured using real‐time qPCR.ResultsLV‐to‐body weight (LV/BW) ratio did not differ between ANP+/−WT and ANP+/−KO fed NS. HS increased LV/BW ratio in ANP+/−WT but not in ANP+/−KO. Gene expression of corin, iNOS, ACE, AT1a, sGC‐α1 and ‐β1 were increased, and NPR‐C was decreased in ANP+/−WT, and unaltered in ANP+/−KO.ConclusionsMaternal ANP ablation appears to protect against the development of CH in the ANP+/− mouse. Compensatory changes in the ANP−/− mother during development may confer salt‐resistance in the ANP+/− mouse later in life, the mechanism of which remains to be elucidated. (Support: Heart & Stroke Foundation of Ontario)Grant Funding Source: Heart and Stroke Foundation of Ontario
The role of Toll‐like receptor 3 in apoptosis of bewo cells hepatitis B virus induced
Authors: Wang Suping , Li Xiaoyan , Song Xiuxia , Feng Liping , Qu Yiqun 
Published: June 2021

ObjectiveThe aim of this study was to explore whether HBVcan replicate in bewo cells, whether HBV can induce apoptosis of bewo cells, and the role TLR3 may play in apoptosis induced by HBV.MethodsPCR was used to detect HBV cccDNA expression of bewo cells cocultured with HBV positive serum. Flow cytometry, tarnsefarse‐mediated dUTP nick end‐labeling were used to detect apoptosis of bewo cells. Flow cytometry was conducted to measure TLR3 protein.ResultsHBV cccDNA can be detected in bewo cells co‐cultured with HBV DNA positive serum for 48 hours. when bewo cells were cocultured with 100μl HBV serum for 24h or 48h, early apoptotic rates, total apoptotic rates and apoptotic indexes were all statistically higher than that in the separate cotemporaneous group (P < 0.001). After PolyI:C prestimulation, early apoptotic rates and total apoptotic rates of the bewo cells co‐cultured with HBV serum for 24h or 48h were significantly decreased(P < 0.05); TLR3 protein's mean fluorescence intensity was statistically higher than that in no‐PolyI:C stimulated group(P=0.02).ConclusionBewo cells which hepatitis B virus can replicate in may be a good model for HBV infection experiments. TLR3 may mediate the apoptosis of bewo induced by HBV.The activation of TLR3 by PolyI: C may inhibit apoptosis of bewo induced by HBV.
The fructose 2,6‐bisphosphate cycle bioswitch and metabolic coordination
Authors: Okar David Anthony 
Published: June 2021

ObjectiveProvide a comprehensive mechanistic description of metabolic regulation that can account for the full spectrum of positive synergistic effects observed while investigating the modulation of hepatic glycolysis as a therapy for diabetes‐associated hyperglycemia.MethodSearch pertinent literature for examples of metabolic coordination including those observed for hepatic fructose 2,6‐bisphosphate (F‐2,6‐P2 and 6‐phosphofructo‐2‐kinase/fructose‐2,6‐bisphosphatase (PFK‐2/FBP‐2) and identify common elements, thereby highlighting mechanisms that coordinate metabolism within the liver and in relation to fuel homeostasis.ResultsThe most obvious regulatory elements common to metabolic coordination are chemical cycles, protein:protein interactions, and translocation between sub‐cellular locations. All of these constitute biological switches and some coordinate across levels of biological organization.ConclusionsThe PFK‐2/FBP‐2 dimer is a bioswitch that along with F‐2,6‐P2 primarily modulates the committing step to glycolysis and coordinately regulates multiple aspects of fuel metabolism at the molecular, cellular, tissue/organ, and systemic levels of biological organization. Funding: VA.
The antidiabetic metformin is toxic to liver mitochondria of Zucker Diabetic Fatty (ZDF) rats: a cellular metabolomic study with 13C‐lactate and carbon 13 NMR
Authors: Baverel Gabriel , Pinteur Claudie , Martin Guy , Gauthier Catherine 
Published: June 2021

Metformin decreases hyperglycemia in diabetic subjects and animals. This effect results in part from the inhibition of hepatic gluconeogenesis. The objective of this study was to investigate in vitro the effect of metformin on hepatic lactate gluconeogenesis by using an innovative approach.Precision‐cut liver slices from adult male ZDF rats were incubated with variously 13C‐labeled lactates (5 mM) or with 5 mM unlabeled lactate plus 25 mM 13C‐bicarbonate. Substrate removal and product formation were measured by enzymatic and 13C‐NMR methods.In slices from fasted lean ZDF rats, metformin dose‐dependently inhibited 13C‐glucose synthesis from [2‐13C]‐lactate (IC50 2.3 mM). The labeling patterns of the glucoses synthesized indicated that lactate gluconeogenesis involved not only the passage of carbons through pyruvate carboxylase and the reversible equilibration of oxaloacetate with fumarate but also the entire tricarboxylic acid cycle.In slices from fed diabetic ZDF rats, 2 mM metformin inhibited lactate consumption, glucose production, the synthesis of all the 13C‐labeled glucose carbons, the production of 13CO2 and reduced the cellular ATP level. By contrast, metformin dramatically increased lactic acid production and ketogenesis.These results strongly suggest that a compound found to be toxic to liver mitochondria in vitro should not necessarily be withdrawn from drug development.
The NO trafficking between dinitrosyl‐iron complexes (DNIC) and E. coli transcriptional factors containing iron‐sulfur clusters
Authors: Yu Steve , Lo Fengchun , Hung Mu‐Cheng 
Published: June 2021

SoxR protein is a transcriptional factor belonged to a member of the Mer family. The native form of the protein is presented as a dimeric structure and each monomer contains a redox‐active [2Fe‐2S] center. The iron‐sulfur cluster of SoxR is responsive to one‐electron oxidation by superoxide and resulted in subsequent expression of soxS gene for the rescue of oxidative stress such as the expression of superoxide dismutase or catalase. In addition to superoxide, SoxR is also sensitive to nitric oxide (NO) and its nitrosylated protein displayed paramagnetic features where its average g value is appeared at 2.03 in EPR. Despite the crystal structures of SoxR protein with and without the nucleic acids have already been unraveled [1], there is still not sufficient structural information to illustrate how the conformational changes mediated by redox chemistry or NO modulation [2] could lead with the transcriptional activation. In this study, we employed the x‐ray absorption spectroscopic methods to determine the structural insights of metal active sites in SoxR under the oxidized, reduced and nitrosylated conditions, respectively. We anticipate clarifying the major clues that control the corresponding gene regulations resulting from oxidative or nitrosative stress.Financial Support: National Science Council (NSC 98‐2627‐M‐001‐004‐) and Academia Sinica in Taipei
Pirlindole analogs as probes of the active site of Monoamine Oxidase A
Authors: Ramsay Rona Ruth , Liu Gu , Westwood Nicholas J 
Published: June 2021

Pirlindole analogs are antidepressants that selectively inhibit mitochondrial monoamine oxidase A. New analogs were synthesized to probe the interaction with the FAD prosthetic group. Previous work (Hynson, R.M.G., Wouters, J. and Ramsay, R.R. (2003) Biochem. Pharmacol. 65, 1867–74) showed that changing the position of the pirlindole nitrogen relative to the N5 of the isoalloxazine ring altered the intensity of the spectral shift at 500 nm. New derivatives masking the pirlindole nitrogen or changing it to C or O provide experimental support for the position of the inhibitor docked in the active site.Supported by INTAS 99‐00433
Expression and purification of active human recombinant cathepsin K in E. coli
Authors: Tha Soe , McConnell Matthew , Berryman Laura , Chmielowski Jennifer , Bordowitz Michael , Liang Kenneth , Lafollette Jessica , Wen Lisa , McConnell Rose 
Published: June 2021

Cathepsin K, a lysosomal cysteine protease, is abundantly expressed in osteoclasts and involved in bone remodeling and resorption. It has been reported that inhibitors of cathepsin K show great potential in the treatment of osteoporosis. The objective of this research is to prepare functional recombinant cathepsin K enzyme which will be used in the development of new cathepsin K inhibitors. Human procathepsin K cDNA has been amplified by PCR and successfully subcloned into pET15b expression vector. Overexpression of the recombinant His‐tagged fusion protein in BL21(DE3)pLysS host cells was found mainly in inclusion bodies. The inclusion bodies were solubilized by 6 M guanidine hydrochloride and the recombinant procathepsin K protein purified using Ni‐NTA affinity column. The purified protein was refolded by dilution followed by dialysis. The procathepsin K protein has been autoactivated and its proteolytic activity assayed by monitoring the increase in absorbance at 405 nm due to hydrolysis of chromogenic substrate, Z‐Phe‐Arg‐pNA. The availability of the functional cathepsin K enzyme will permit a drug discovery program for development of newer effective therapy for the treatment of osteoporosis in the near future.Supported by National Cancer Institute at NIH (Grant No. 3R15CA086933‐04 and 3R15CA086933‐04A2S1)
Genetic Polymorphism of lipoprotein lipase, Beta 3 Adrenergic Receptor and Plasma Cell Membrane Glycoprotein and The Susceptibility to Dyslipidemia
Authors: El Din Hemimi Neveen Salah , Tawfik Lamis M , Elhannan Sahar Elsaid 
Published: June 2021

BackgroundDyslipidemia is one of the most common disorders in the modern society to the extent that it is the major determinant of the degree of excessive morbidity and mortality.AimIdentification of subject at risk of dyslipidemia in order to carry more effective interventions to prevent the development or progression of the associated complication such as cardiovascular diseases.Material and MethodsThe study group consisted of 202 individual with different lipid profiles. Individuals were genotyped for LPL Hind III, LPL S447X, β 3 ADR and PC1 by using PCR‐RFLP. Statistical analysis was done by using SPSS program.ResultsCarriers of H+/H+ genotype were at significant high risk of developing dyslipidemia manifested by high triglycerides, high LDL and low HDL (OR=1.9, 2.3 and 2.0) respectively. SS genotype was associated with significant increase risk of total cholesterol (OR= 2.7). C allele of β 3 ADR was a risk factor for low level HDL (OR= 2.3). Carriers of Q allele of PC1 were at significant risk of developing atherogenic phenotype manifested by low level HDL and high level LDL (OR=2.4 and 5.1) respectively. Furthermore combined analysis revealed that carrier of two or more of the previous haplotypes were at the highest risk of developing dyslipidemia.ConclusionH+, S, C and Q alleles were risk factors of developing atherogenic lipid profile.Source of fund: Ain Shams University.
Marinobufagenin is an upstream modulator of Gadd45a stress signaling in cytotrophoblast cells
Authors: Uddin Mohammad Nasir , Horvat Darijana , DeMorrow Sharon , Agunanne Enoch , Puschett Jules B 
Published: June 2021

ObjectiveMarinobufagenin (MBG), a cardiotonic steroid, is increased in preeclampsia (PE). Gadd45a is involved in the pathophysiological changes observed in PE. Cytotrophoblast (CTB) cells play an important role in placental development.Study aimto elucidate the role of MBG in Gadd45a signaling in CTB cells.MethodsCTB cells were treated with various concentrations of MBG (0, 0.1, 1, 10 or 100 nM). Gadd45a expression and p38 phosphorylation were measured by immunoblotting. The degree of cell cycle arrest was determined by FACS analysis. sFlt‐1 and 8‐isoprostane were measured by EIA. Gadd45a expression was inhibited in CTB cells via siRNA‐mediated knock down.ResultsGadd45a expression and p38 phosphorylation were significantly upregulated in the 1, 10 and 100 nM MBG‐treated cells compared to vehicle‐treated cells. 0.1 nM MBG had no effect. There was a significantly higher percentage (80%) of cells in the G0/G1 phase in the cells treated with ≥1.0 nM MBG compared to basal and to 0.1 nM treated cells (55%). The application of MBG ≥1 nM significantly increased the secretion of sFlt‐1 and 8‐isoprostane. The RNAi‐mediated inhibition of Gadd45a attenuated MBG‐induced CTB cell stress signaling.ConclusionsMBG caused upregulation of p38 phosphorylation and cell cycle arrest, and increased the secretion of sFlt‐1 and 8‐isoprostane. These alterations occurred as a result of Gadd45a stress signaling.
A Novel Mono‐carbonyl Analogue of Curcumin Induces Apoptosis by Activating ER Stress in Non‐Small Lung Cancer Cells
Authors: Xiao Jian , Liang Guang , Huang Yi , Li Xiaokun , Studer Elaine , Zhou Huiping 
Published: June 2021

Non‐small lung cancer (NSCLC) is one of the most common cancers worldwide. Curcumin, a well‐known food additive and constituent of traditional medicine, has been used to treat several different cancers including NSCLC in clinical trials. However, its poor bioavailability and pharmacokinetic profiles due to its instability under physiological conditions have limited its application in anti‐cancer therapies. In this study, we synthesized a novel mono‐carbonyl analogue of curcumin, (1E,4E)‐1,5‐bis (2‐methoxyphenyl) penta‐1,4‐dien‐3‐one (B63), and examined its proapoptotic effect in NSCLC cells.MethodsH460 or H358 cells were treated with different amounts of curcumin and B63 for various time periods. Cell proliferation and apoptosis were measured by MTT assay and flow cytometry. The activation of ER stress was determined by Western blot analysis. The ER Ca2+ content was measured using the fluorescent Ca2+ indicator fura 2/AM. Lentiviral shRNA was used to knock down the ER stress responsive gene CHOP.ResultsB63 had better bioavailability and more potent proapoptotic activity than curcumin. B63, not curcumin, significantly induced depletion of ER Ca2+ store and activation of the unfolded protein response (UPR). Downregulation of CHOP expression inhibited B63‐induced apoptosis.ConclusionActivation of the UPR represents a key molecular mechanism underlying B63‐induced apoptosis in NSCLC cells.
p19INK4d function linked to DNA repair relies on sequential phosphorylation
Authors: Marazita Mariela C. , Ogara M. Florencia , Sonzogni Silvina , Pignataro Omar P. , Cánepa Eduardo T. 
Published: June 2021

p19INK4d(p19), a member of INK4 cell cycle inhibitors, plays a role in DNA repair and becomes phosphorylated upon DNA damage. We aimed to evaluate first the involvement of predicted p19 phosphorylation sites in its DNA repair activity, second to demonstrate the phosphorylation event in those sites, and last to establish the subcellular localization of the phosphorylation process. p19 mutants were made replacing Ser or Thr by Ala at S13, S66, S76, T89, T141. Wi‐38 cells were transfected with the mutants and cell cycle arrest ability by 3H‐timidine incorporation, DNA repair capacity by UDS and apoptosis by caspase‐3 activity were assayed. Although none of the mutated sites are involved in cell cycle arrest capacity, S76 and T141 are necessary for p19 DNA repair function. Mutation of S76 and T141 to glutamic acid to mimic phosphorylation leads to the recovery of the function. To evaluate in vivo phosphorylation of these mutants we performed metabolic labeling with 32P‐orthophosphate. While S76A entirely supressed phosphorylation, T141A didn't completely abrogated. The mutant S76E retrieves the phosphorylation potential. p19 translocates into the nucleus after genotoxic treatment. We found that p19 phosphorylation begins in the citoplasm. These results show p19 is sequentially phosphorylated starting in the cytoplasm and being necessary for exerting an effective DNA repair function.Research supported by ANPCyT.
Epigenetic marks of Insulin‐related transcription factor network by DNA methylation
Authors: Ito Shuhei , Sato Shinya , Yoshioka Tetsuya , Ohgane Jun , Tanaka Satoshi , Yagi Shintaro , Shiota Kunio 
Published: June 2021

Numerous data of transcription factors for Insulin gene expression has been accumulated and used to characterize the Insulin‐producing cells induced from pluripotent stem cells. DNA methylation is a pivotal mark of epigenetic regulation of genes. There are numerous tissue‐dependent differentially methylated regions (T‐DMRs). DNA methylation profiles, which are consisting of methylated and unmethylated T‐DMRs, are unique to the cell types. Here, we analyzed DNA methylation profile of insulinoma MIN6 cell line by genome‐wide DNA methylation analysis method for T‐DMR analysis (D‐REAM). D‐REAM analysis revealed that MIN6 cells have unique DNA methylation profile distinctive from that of insulin non‐expressing cells including embryonic stem cells. Several transcription factors which are involved in Insulin 2 gene expression had T‐DMRs uniquely hypomethylated in MIN6 cells. Interestingly, main part of the Insulin transcription factor network, which involves transcription factors regulating other transcription factors, are under the epigenetic regulation by DNA methylation. Thus, Insulin‐expressing cells had a distinctive DNA methylation profile from others. DNA methylation profile is powerful to characterize the cells for regenerative medicine and the T‐DMR information will provide critical information for evaluating the Insulin‐producing cells.This work was supported by NEDO and NIBIO.
Regulation of CMO1 and CMO2 expression during mouse embryonic development
Authors: Wassef Lesley , Kuhn Elizabeth , Kim Youn‐Kyung , Quadro Loredana 
Published: June 2021

β‐carotene‐15, 15′‐oxygenase (CMO1) and β‐carotene‐9,10′‐oxygenase (CMO2) are the only enzymes known to generate vitamin A from β‐carotene. Both are expressed in mouse and human embryonic and extraembryonic tissues, suggesting a role for β‐carotene as a localized source of vitamin A for developing tissues. Studies indicate that β‐carotene cleavage activity is regulated by nutrient availability in adult tissues. Here we investigate whether nutrient availability may regulate the expression of CMO1 and CMO2 during mouse embryogenesis.Wildtype and CMO1−/− pregnant females were administered β‐carotene by IP injection at 13.5 dpc and sacrificed one day later to collect maternal serum and liver, embryos, yolk sac and placenta. This procedure leads to detectable amounts of β‐carotene in these tissues. Real‐time PCR analysis demonstrated that, in maternal liver, CMO1 expression is down‐regulated while CMO2 is up‐regulated with increasing amounts of β‐carotene. Furthermore, HPLC analysis revealed that administration of β‐carotene also altered maternal serum retinol levels, suggesting the existence of highly regulated mechanisms to ensure that adequate amount of vitamin A are available to the embryo. Experiments are ongoing to analyze the changes in CMO1 and CMO2 expression in embryonic tissues in response to maternal β‐carotene availability and vitamin A status.Grant Funding Source: NIH 5R01HD57493.