bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cobalt Nanoparticles Cause Major Platelet Aggregation In Vitro While Chromium Nanoparticles Induce Platelet Lysis In Vitro.
Authors: Pasieka Paweł Melchior , Marycz Krzysztof , Radomski Marek , Santos-Martinez Maria Jose , Pękala Przemysław Andrzej , Tomaszewska Iwona , Tomaszewski Krzysztof Andrzej 
Published: April 2020

IntroductionThe usage of artificial joint prostheses has recently gained significant popularity, with roughly two million total hip, knee and spinal arthroplasties performed annually worldwide. However, the presence of prostheses in organism is associated with significant release of nanoparticles (NPs). When in the bloodstream they can affect various blood components, such as platelets. Despite that the problem of NPs’ influence on platelets function has not been thoroughly analyzed. Therefore the aim of our study was to analyze the impact of Cr and Co particles on thrombocytes in vitro with usage of novel quartz crystal microbalance with dissipation (QCM‐D) methodology.Materials and methodsThe light transmission aggregometry, flow cytometry and QCM‐D were utilized to assess the ability of NPs to cause activation and aggregation of platelets. Transmission electron microscopy (TEM), scanning electron microscopy and optical and immunofluorescence microscopy were used to confirm the outcomes given by QCM‐D.ResultsCompared to the controls under flow there was a significant change of frequency and dissipation in platelet‐rich plasma incubated with Co 28nm, CoO 50nm, Co2O3 50nm, Co3O4 30–50nm, Cr 35–45nm, Cr2O3 60nm NPs (at concentrations of 5, 2.5, 1, 0.5 μg/mL). The NPs induced platelet aggregation even at the lowest concentrations. Other utilized modalities confirmed aggregation of thrombocytes. Furthermore, TEM showed that Cr NPs induce swelling and lysis.ConclusionOur study indicates that both Cr and Co NPs influence thrombocytes function in vitro. Also two different underlying mechanisms of platelet aggregation as Cr NPs cause swelling and lysis while Co particles induce typical aggregation. We conclude that any physician should take into consideration monitoring level of Cr and Co NPs of patients having Co‐Cr prosthesis.
Tissue‐specific Humanized Models Expressing Human UGT1A1
Authors: Mennillo Elvira , Yang Xiaojing , Weber André , Chen Shujuan , Maruo Yoshihiro , Tukey Robert H. 
Published: April 2020

The human UDP‐glucuronosyltransferases (UGTs) represent an important family of drug‐metabolizing enzymes. Among the nine UGT1A proteins, UGT1A1 has been the most extensively studied. UGT1A1 is the sole enzyme responsible for the glucuronidation of bilirubin and is involved in the conjugation of endogenous and many exogenous substances, including pharmaceutical drugs. Although the liver is considered a major organ for detoxification, intestinal UGT1A1 is also an important contributor for drug clearance. In order to investigate the role of both intestinal and hepatic human UGT1A1, we generated humanized strains that express only the UGT1A1 gene in either liver (hAlb1A1) or small intestine (hVil1A1). The two models were characterized by demonstrating hepatic and intestinal tissue specific expression of human UGT1A1. Body weight (BW) and total serum bilirubin (TSB) levels were monitored during different developmental stage. Both humanized models exhibited similar BW and normal TSB values in the adult stages. Differences between the two animal models were observed in the neonatal stage where hVil1A1 pups presented significantly lower TSB levels than hAlb1A1 mice, confirming that the intestinal expression of UGT1A1 is crucial in preventing bilirubin toxicity during development. Of importance is the finding that we have new humanized UGT1A1 mouse models to examine tissue‐specific glucuronidation. Given the importance of the intestinal tract in xenobiotic activation, metabolism and detoxification, we subsequently have isolated intestinal crypt organoids from these same animal models. The use of these in vivo models and their in vitro complementary primary cell models provide important tools for studying intestinal and hepatic toxicity of drugs and environmental toxicants that are metabolized by UGT1A1.Support or Funding InformationSupported by National Institutes of Health grants GM126074 and ES010337
Rates of Synthesis of Individual Proteins in Muscles of Old Mice: A Heavy Water Approach.
Authors: Owen Euan , Hammond Dean E. , Staunton Caroline , Franco Catarina , Prescott Mark , Brooks Susan V. , Jackson Malcolm J. , Lau Edward , Beynon Rob J. , McArdle Anne 
Published: April 2020

Age‐associated loss of muscle mass and function is a major factor in the development of frailty, but the mechanisms responsible for this loss are unknown. Unquestionably, muscle atrophy is the consequence of changes in the balance between protein synthesis and breakdown. However, the nature and extent of these changes in old age is unclear, with some studies reporting reduced basal muscle protein synthetic rate in the aged compared with younger people and others suggesting no difference. Whole organism heavy water ([2H2]O) dynamic labelling of proteins, followed by LC‐MSMS, provides a significant improvement in analysis of protein turnover, allowing the rates of synthesis and degradation of both slow and fast turnover muscle proteins to be accurately modelled. The aim of this study was to use a [2H2]O labelling approach to determine the synthesis rates of individual proteins in the gastrocnemius muscles of adult and old mice.Ten adult and ten old male mice were primed with an intraperitoneal bolus injection of saline, prepared with 99% [2H2]O and subsequently, provided with 8% (v/v) [2H2]O in the drinking water, for up to 60 days. The sampling interval was distributed to obtain sufficient data points to cover both high and low turnover proteins. Muscle proteins were extracted and digested using standard procedures for analysis on a high‐resolution mass spectrometer (Thermo Q‐Exactive) and the raw data files were processed to allow protein identification using Crux‐tide (
McIlwain et al, 2014). The software RIAna (:
https://github.com/Molecular‐Proteomics/riana) was then used to track time dependent incorporation of [2H] into tryptic peptides, yielding synthesis rates of proteins to be determined.Muscle mass was significantly reduced in the old mice and preliminary data demonstrated little change in the rate of synthesis of a large proportion of identified muscle proteins between adult and old mice. Several individual proteins showed a marked decrease in protein synthesis rates in muscles of old compared with adult mice, particularly structural proteins. Other proteins, particularly redox‐responsive proteins demonstrated an increased synthesis in muscles of old mice. Proteomic analysis of unlabelled muscle protein pools and RNASeq data will be integrated with the turnover data to further interpret these findings.This study was supported by the University of Liverpool, MRC and NIA (AG051442).
A Striking Bone Phenotype in the Familial Danish Dementia (FDD) Tg+ Mouse
Authors: Suryadevara Vidyani , Krehbiel Connor J. , Vidal Ruben , Willis Monte S. 
Published: April 2020

Familial Danish Dementia (FDD) is an autosomal dominant neurodegenerative disease caused by a 10‐nucleotide insertion mutation in the BRI(2) gene (aka ITM2B) resulting in a frame‐shift. The resulting Danish amyloid precursor protein (ADanPP) is then cleaved an amyloidogenic peptide that deposits in the brain and cerebral vessels. FDD in patients results in a range of clinical defects, including cataracts, hearing defects, cerebellar ataxia, slurred speech, gait disturbances, psychosis, and dementia. With emerging evidence that human bone defects are associated with neurodegenerative disease, we phenotyped bone in 12‐month FDD Tg+ mice. FDD Tg+ mice exhibit abnormal grooming by 9 months and have Ab plaques and behavior deficits by 12 months, paralleling mechanisms in Alzheimer disease. At 12 months of age, femurs from FDD Tg+ mice were assessed by mCT (SkyScan 1172, 9 mm resolution) in the distal metaphysis (1 mm) and mid‐diaphysis regions for trabecular and cortical areas, respectively. Both male and female femurs exhibited significantly reduced bone volumes (44%) vs. age‐, strain‐matched wildtype mice; male FDD Tg+ femurs showed a 24% reduction in bone mineral density.Trabecular boneFemales FDD Tg+ femurs had a 24% reduction in the trabecular thickness (vs. controls), whereas male FDD Tg+ femurs exhibited a 24% increase in the trabecular separation and 40% reduction in the trabecular number (vs. controls).Cortical boneFemale FDD Tg+ femurs had a 15% reduction in bone marrow area and 8.6% endocortical thickness compared to controls. Male FDD Tg+ femurs had reduced total cross‐sectional area and endocortical thickness by about 8%, a 15% reduction in the marrow area, along with a reduced periosteal thickness and increased cortical thickness vs. controls. These results indicate that FDD Tg+ mice have significant bone phenotypes in parallel with previously observed brain defects, suggesting therapeutic interventions may want to consider how neurodegeneration might impact other critical organs in the body such as bone.Support or Funding InformationLilly Foundation/Indiana Center for Musculoskeletal Health/ IU School of Medicine, Physician Scientist Initiative, Scientific Research Initiative
Mapping Circuit Connections of Brainstem Respiratory Regions with Retrogradely Transporting Viral Vectors in Adult Transgenic Rats
Authors: Sheikhbahaei Shahriar , Gamarra Julio , Chopra Varuna , Zhang Ruli , Smith Jeffrey C. 
Published: April 2020

In mammals, breathing is a constantly regulated key motor behavior that is generated by a rhythmic neural process in the brainstem. The preBötzinger complex (preBötC), a medullary region critical for generating the breathing rhythm, has been proposed to have circuit connections ipsilaterally and contralaterally to functionally distinct neighboring medullary regions, such as the respiratory Bötzinger complex (BötC), the rostral and caudal Ventral Respiratory Groups (rVRG/cVRG), and retrotrapezoid nucleus (RTN) for coordination of the rhythmic activity of multiple respiratory neuronal populations. Because breathing is integrated with other volitional and physiological behaviors, it is proposed that brainstem respiratory circuits, especially within the preBötC, act as critical nodes for control of breathing, and may have connections to neurons in other regions throughout the brain involved in motor control and behavioral integration such as motor cortex for volitional control of breathing or other aspects of motor integration. However, the axonal projections of preBötC neurons throughout the brain, and the input connections from other brain regions have not been mapped. In the present study, we therefore performed brain‐wide mapping employing adult transgenic rats expressing Cre recombinase under control of the glutamic acid decarboxylase 1 (GAD1) promoter to specifically target GABAergic inhibitory neurons. To map input connections to the preBötC, we used combinations of Cre‐dependent and Cre‐independent retrogradely transporting viral vectors (rAAV2‐retro) driving expression of different fluorescent proteins to allow connections of GABAergic neurons to be differentiated from connections by other neuron types. This approach allows for targeted mapping of local circuit neurons and their axonal projections as well as retrograde labeling of neurons with long‐range input connections. With this approach, we have obtained evidence of retrograde GABAergic and non‐GABAergic neuronal labeling at multiple levels of the brainstem (including BötC, rVRG, RTN, and pontine Kölliker‐Fuse/Parabrachial nuclei) and numerous suprabrainstem regions including cortical and subcortical regions. Our results indicate that various constructs of rAAV2‐retro in combination with specific Cre transgenic lines are effective tools for brain‐wide mapping of input connections to circuits in functionally defined respiratory regions, including providing genetic access to specific populations of projection neurons.Support or Funding InformationIRP of NIH, NINDS
SPARC: Correlation between Patient Characteristics in Gastrointestinal Evaluation using Principal Component Analysis
Authors: Dabiri Yaghoub , Gregersen Hans , Kassab Ghassan 
Published: April 2020

Introductionanorectal function is complex. Assessment of anorectal physiology and diagnostics for anorectal disorders face problems due to inconsistent results with different technologies and huge inter‐subject variability. This limits development of therapeutic strategies. Devices used to evaluate anorectal function include Fecobionics, EndoFLIP, Anorectal manometry and Ultrasound. The goal of this study was to find correlation in patient characteristics based on data obtained with these measurements.Methodsthe four tests mentioned above were used to evaluate anorectal function in normal subjects and patients with fecal incontinence (FI) or constipation. The data were cleaned by replacing unknown values with the average of measurements, and they were standardized. In total there were 90 measurements and 35 features. The collected data included sex, age (60.0±14.3 years), weight (59.0±9.0 kg), height (1.6±0.1m), FI score (16.8±12.8 with normal values =< 5), constipation score (6.5±4.4 with normal values =< 8), anorectal maximum pressure (46.9±10.1 mmHg), resting pressure (61±25.8 mmHg), expel time (46.3±63.2s), and absence or presentation of rectoanal inhibitory reflex (RAIR), etc. Principal component analysis (PCA) was used to find the contribution of each reduced feature in variance of data and the correlation between features, utilizing singular value decomposition. We used Python and its libraries.Results and Discussionthe variance in data was mostly explained using 25 PCs. The percentage of cumulative variance explained by up to the 1th, 25th and 35th PC was 15%, 98% and 100%, respectively. The PCs provided insights about the relation between features. For example, according to the first PC the maximum pressure, resting pressure, maximum tolerance and age were positively correlated. Also, weight, absence of RAIR and constipation score were positively correlated. However, the latter group of parameters were negatively correlated to the former group. Because the number of patients used in this study were limited results should be interpreted with caution. However, more data are currently being collected, which will be included in our analysis. We plan to extend our analysis to cluster patient data using machine learning.Conclusionsin this study we analyzed data gathered in anorectal evaluation using PCA. Data obtained provided insights about relation between characteristics of the patients. Analysis of data obtained in anorectal evaluation will help to better understand factors involved in anorectal and other gastrointestinal disorders, which could lead to better preventive, diagnostic and therapeutic strategies.Support or Funding InformationNIH SPARC fundingAcknowledgementsNIH SPARC funding.
Anatomical reorganization within the hand and forelimb of Perodicticus potto
Authors: Boettcher Marissa Lauren , Dickinson Edwin , Leonard Kaitlyn , Herrel Anthony , Hartstone-Rose Adam 
Published: April 2020

Historical behavioral accounts note extraordinarily high grip strength in Perodicticus potto. Anatomical analysis of the potto’s hand and forelimb reveal several specializations that may facilitate this enhanced gripping ability, including a specialized vascular bundle (the retia mirabilia) within the wrist, a hyper‐abducted pollex, and a highly reduced second digit. The myological correlates of grip strength (i.e., the contractile potential of the hand and wrist musculature) of P. potto, however, have yet to be quantitatively evaluated. The current study contrasts the myology of the hand and forearm muscles within P. potto to a series of closely‐related strepsirrhines (Nycticebus coucang, Otolemur crassicaudatus, Hapalemur griseus, and Eulemur mongoz), as well as comparing the forearm musculature to a broader (n= 41) primate sample. Despite reports of the potto’s high grip strength, our findings demonstrate relatively below‐average force capacity (relatively low physiological cross‐sectional area) within the digital flexors of this species, which suggests that the gripping ability of this species may be less extraordinary than previously hypothesized. In addition to this observation, we report a number of unusual myological features within the potto, including the largest brachioradialis (an elbow flexor) of any taxon within our sample. Even more surprisingly, we identified an independent extensor indicis within P. potto, despite the vestigial nature of the second digit, and the absence of this muscle within ~25% of other taxa within our sample ‐ all of which possessed a more substantial second digit than the potto. These data underscore the curious nature of the potto’s antebrachial musculature, but suggest that adaptations within the potto’s forelimb may be more related to maximizing endurance grip potential than towards increasing myological force.Support or Funding InformationNational Science Foundation (IOS‐15‐57125 and BCS‐14‐40599)
Krill Oil Addition to 8‐weeks of High‐fat Diet Enhances mTORSer2448 Phosphorylation in Rat Heart
Authors: Butcko Andrew , Andraka John , Sharma Naveen 
Published: April 2020

Krill oil (KO) derived from shrimp‐like crustaceans, has been marketed as a substantial source of dietary long chain omega‐3 (LC n‐3) polyunsaturated fatty acids, particularly rich in eicosapentaenoic acid (EPA; 20:5 n‐3) and docosahexaenoic acid (DHA; 22:6 n‐3). Consumption of EPA and DHA has been linked to the reduction of cardiovascular disorders including coronary heart disease and myocardial infarction, however the cellular mechanism for these cardioprotective benefits is unclear. The mechanistic target of rapamycin (mTOR) is a Ser/Thr kinase that acts a nutrient sensor, subsequently controlling various key cellular processes including cell growth, cell survival, and protein synthesis. Diets high in saturated fat content have been associated with myocardial dysregulation through alteration of the mTOR signaling pathway, suggesting that interventions that ameliorate mTOR signaling may counter these deleterious effects in the heart. The goal of the current study is to determine the effect of krill oil supplementation to a high saturated fat diet on mTOR signaling in cardiac muscle. Male Sprague‐Dawley rats (approximately 12 months‐old), were assigned by mass to one of three isocaloric dietary groups for 8 weeks: control diet group (CON; 17% saturated fat, 54% carbohydrate, 29% protein; n=8), a high saturated fat group (HF; 42% saturated, 43% carbohydrate, 15% protein; n=12), and a high fat group with krill oil supplementation (KO; 34% saturated fat, 8% krill oil, 43% carbohydrate, 15% protein; n=12). All rats were given ad libitum access to food and water. Body weights and food consumption was recorded weekly. After 8 weeks of dietary intervention and an overnight fast, blood and hearts were collected from anesthetized rats during a terminal surgery. Western immunoblot analysis was used to determine phosphorylation levels of key mTOR signaling proteins including mTORSer2448, p70S6KThr389, 4E‐BP1Thr37/46, eIF4ESer209, and eIF2αSer51. There was no difference in body mass between dietary groups after 8 weeks of treatment. There was a significant increase (p<0.05) in phosphorylation of mTORSer2448 in the KO group when compared to the HF and CON groups. Phosphorylation of downstream targets of mTOR was not different between groups. Eight weeks of KO supplementation can enhance phosphorylation of mTORSer2448, however the cardioprotective impact of this remains unknown. Further research is needed to examine how KO consumption can affect other key downstream mTOR‐related targets associated with improved cardiovascular health.Support or Funding InformationResearch was supported by intramural grants from the College of Health Professions and the Office of Research and Graduate Studies at Central Michigan University.
Overexpression, Purification, and Preliminary Crystallographic Analysis of the Archaeosine Synthase GAT‐QueC
Authors: Susanto Albert , Honberger Matthew , Murphy Ryan , Bayooz Susan , Yi Stuart , Swairjo Manal A. 
Published: April 2020

Archaeosine (G+) is a highly modified nucleosides found exclusively at position‐15 in the dihydrouridine loops (D‐loop) of archaeal tRNAs where it contributes to tRNA structural stability. In a late step in G+ biosynthesis, the precursor nucleoside 7‐cyano‐7‐deazaguanine (preQ0) is inserted into tRNA by the enzyme tRNA‐guanine transglycosylase (arcTGT) before conversion to G+ by archaeosine synthases that vary among archaeal phyla: ArcS in Euryarchaeota, and QueF‐like or GAT‐QueC in Chrenoarchaeota. GAT‐QueC is a two‐domain enzyme harboring an N‐terminal glutamine amidotransferase (GAT) domain, and a C‐terminal domain homologous to QueC, the enzyme that produces preQ0. Consistent with its 2‐domain structure, GAT‐QueC functions in vivo as both a preQ0 synthase and an amidinotransferase, generating G+‐modified tRNA. However, it remains unknown whether the GAT domain acts on preQ0 before or after its insertion into tRNA by arcTGT. Here we describe overexpression, purification and preliminary biochemical and crystallographic characterization of GAT‐QueC from Sulfolobus tokodaii (StGAT‐QueC). Analytical gel filtration chromatography and electrophoretic mobility shift analysis reveal that StGAT‐QueC is a monomeric protein with molecular weight of 55 kDa and no affinity for tRNA, suggesting that the enzyme acts on free preQ0, not preQ0‐modified tRNA. High throughput crystallization of the apoenzyme (and in the presence of preQ0) using the vapor diffusion method yielded crystals that grew in 21 days from solutions containing methyl pentanediol and sodium acetate, and diffracted to 5.8 Å without cryo protection. The crystals belong to cubic space group F23, exhibit unit cell length=292 Å and contain 4 protein molecules per asymmetric unit.Support or Funding InformationNIGMS grant GM110588 and GM132254 to M.A.S., and The California Metabolic Research Foundation (SDSU).