bluuebunny/crossref_metadata_2025_split huggingface dataset:
Case Report: A Unique Unilateral Variation of a Divided Extensor Digitorum Muscle in a Subject with Bilateral Absence of Extensor Indicis
Authors: Schettler Stephen P. , Lister James P. , Miller Joseph M.A. 
Published: April 2020

Reports of variations in the extensor digitorum muscle have been documented since at least 1813. These variations have fallen into 6 groups: (1) deficiencies in tendon number – usually the tendon to the 5thdigit being absent, (2) doubling of tendons to digits, (3) difference in number of muscle bellies, (4) presence of tendinous slips that join other muscles, (5) presence and arrangement of intertendinous connections, (6) presence of separate muscle bellies for each tendon, and (7) accessory muscle bellies.The present case report describes a unilateral variation of the extensor digitorum muscle, observed in a 73 year‐old Caucasian male with bilateral absence of the extensor indicis muscles. During anatomical dissection of this individual, we observed that the extensor digitorum muscle on the subject’s right side appeared to be divided into two distinct muscle masses. The lateral muscle mass (1 cm wide at its thickest point and 11 cm in length) originated from the medial aspect of the extensor carpi radialis brevis and provided a tendon (20 cm in length) to the index finger. The medial muscle mass had the usual origin to the lateral epicondyle via the common extensor tendon and provided tendons to the third and fourth digits. Interestingly, the extensor indicis muscles were also absent bilaterally.While cases of absent extensor indicis muscles have been reported previously, we are not aware of any report in the literature that describes a divided extensor digitorum muscle or such a divided extensor digitorum muscle occurring in the presence of bilaterally missing extensor indicis muscles. Indeed, to our knowledge, this is the first such report of this occurrence. Thus, to the 6 groups of variant types commonly recognized for extensor digitorum as listed above, we now add a 7th new category of “divided muscle mass” (which in our case is “bipartite”, but future studies may discover other arrangements). Developmental and functional considerations of this variation will be further discussed.Support or Funding InformationDepartment of Pathology and Laboratory Medicine, David Geffen School of Medicine at UCLA
seededclustR: Using Cluster Analysis of Mitochondria to Improve Cell Segmentation and Automated Quantification of mtDNA in Smooth Muscle Cells.
Authors: dos Santos Anita , Ng Irvin , Poburko Damon 
Published: April 2020

IntroSegmentation of fluorescently labelled cells is essential to pipelines of automated image analysis. Identifying regions of interest (ROIs) around labelled nuclei is relatively easy using tools available in ImageJ/Fiji, but determining the bounds of cells can be much harder. We asked if fluorescently labelled mitochondria could be used to improve the automated generation of cellular ROIs.MethodThe perimeter coordinates of nuclear ROIs and the centroid coordinates, area, mean intensity, angle, and solidity of ROIs around MitoTracker Orange (MTO)‐labelled mitochondria were generated in ImageJ and exported to R. Using the nuclei as seeds for a recursive k‐Nearest Neighbour search extending from each nucleus, MTO particles were assigned to their nearest‐neighbouring cells. A concave hull was created around MTO particles assigned to each cell using the concaveman package. We determined the sensitivity of this algorithm by comparing the accuracy of the generated concave hulls to the corresponding hand‐drawn ROIs, and ROIs from Voronoi expanded distance mapsResultsWhen we compared seededClustR ROIs to those drawn by hand, the Intersection‐Over‐Union (IOU) was 68 ± 8% (mean ± SD), Precision was 88 ± 6% and Recall 95 ± 3%. In contrast, Voronoi expansion of nuclei resulted in the IOU of 46 ± 18%, Precision of 83 ± 17%, and Recall of 91 ± 11%. Optimal IOU and Recall were observed when each MTO particle was compared to its 7–10 nearest neighbours. IOU and Recall were maximized using recursive steps of 40–200 μm, while Precision decreased from 94% to 88% as step size increased from 7–40 μm. Unexpectedly, extending seededClustR to an n‐dimensional kd‐tree calculation that included varying weights of MTO particle mean intensity, area, standard deviation, angle and solidity reduced cell tracing accuracy. Similarly, using Wilks’‐test to identify outliers decreased IOU by 17 percentage points (pp) and Recall by 20 pp, though it increased Precision by 6 pp. Finally, we tested whether if the more accurate cell ROI from seededClustR versus Voronoi ROIs could improve our ability to detect changes in optical measures of mtDNA in A7r5 smooth muscle cells treated with angiotensin II, 5,6‐dideoxycytidine, and H2O2, where results showed promising gains.ConclusionseededClustR is a generalizable algorithm that can use simple XY coordinates of puncta‐like features of cells to generate cellular ROIs around labelled nuclei with measures of accuracy that exceed simple dilation of nuclear ROIs. This approach should facilitate improved precision in automated screens of cellular phenotypes.Support or Funding InformationNatural Sciences and Engineering Research Council of Canada
Role of Wnt5a in Switching Macrophage Phenotype in the Tumor Microenvironment
Authors: Hazen Alyx T. , Coschigano Karen T. , Malgor Ramiro 
Published: April 2020

In 2018, cancer was the second leading cause of death in the United States. Past research has demonstrated the importance of macrophages in both the tumor microenvironment and in cancer progression. Macrophages are specialized immune cells that are differentiated into either the M1 phenotype (pro‐inflammatory/tumor‐suppressing) or M2 phenotype (anti‐inflammatory/tumor‐promoting), depending on the signals in the tumor microenvironment. Research has demonstrated that the expression of Wnt5a, a secreted glycoprotein, is correlated with a cancer’s level of aggression. Additionally, evidence shows that the treatment of M1 macrophages with Wnt5a increased the expression of IL‐10, a molecule expressed by M2 macrophages and associated with the progression of cancer. The objective of this study is to investigate the effect of exogenous Wnt5a on the expression levels of M1/M2 phenotypic markers. The hypothesis is that Wnt5a facilitates the switching of macrophages from the M1 to M2 phenotype through the induction of IL‐10. This would support the idea that Wnt5a functions as a tumor promoter and potentially contribute in the progression of cancer.To test this hypothesis, THP‐1 and human peripheral blood monocyte derived macrophages were evaluated for expression of the M1 markers CD80 and IL‐6 and the M2 markers CD163 and IL‐10 using flow cytometry and RT‐qPCR either with or without Wnt5a treatment.Our preliminary results confirm that, without Wnt5a treatment, M1 macrophages strongly expressed Wnt5a and, further, demonstrate that, after treatment with exogenous recombinant Wnt5a, the expression of M2 markers, including IL‐10, increased, while the expression of M1 markers decreased. Results from this study lead us to speculate that Wnt5a could play a role in switching macrophage phenotype from M1 to M2 and thus determining the overall tumor behavior. Furthermore, the manipulation of Wnt5a signaling could potentially be an attractive strategy in the development of more effective cancer treatments.Support or Funding InformationThe John J. Kopchick MCB/TBS Undergraduate Student Support Fund (Ohio University) andThe Provost Undergraduate Research Fund (Ohio University)
Characterization of the Inhibition of Allelic Variants of Cytochrome P450 2A6 with trans‐Cinnamic Aldehyde
Authors: Riehlman Emily , Mogstad Emily , Harrelson John , Chan Jeannine 
Published: April 2020

Cigarette smoking and nicotine addiction continues to be a leading cause of preventable death. The enzyme cytochrome P450 2A6 (CYP2A6), which is highly polymorphic with over forty known alleles, is the primary enzyme that catalyzes the initial oxidation of nicotine in humans. Studies have shown that trans‐cinnamic aldehyde (tCA) is a mechanism‐based inactivator of this enzyme and is therefore a potential smoking cessation aid. However, the inhibitory effectiveness of tCA with the gene products of the different alleles is unknown. The goal of the work presented was to characterize inhibition of the CYP2A6 variant resulting from the *23 allele by tCA. This allele results in a point mutation (R203C). Site‐directed mutagenesis was used to engineer CYP2A6*23 from a truncated form of the human CYP2A6*1 gene. The recombinant protein was expressed in E. coli and purified. Using ligand‐binding titrations, we found that the Kd for tCA binding to CYP2A6*23 was 7.1 μM, whereas the Kd value for CYP2A6*1 was 10.6 μM. Consistent with previous reports, CYP2A6*23 had lower activity than that of CYP2A6*1; however the relative inhibition of both CYP2A6*23 and CYP2A6*1 by 80 μM tCA was similar. Therefore, while tCA has a slightly higher affinity for CYP2A6*23, the change to a cysteine at position 203 does not significantly affect the effectiveness of tCA as an inhibitor. So tCA could still be a potential smoking cessation aid for individuals with the CYP2A6*23 allele.Support or Funding InformationNational Institute on Drug Abuse Award Number 15DA042341; Pacific University Faculty Development Grant
Investigating the Molecular and Functional Impact of a Novel DLG2/PSD‐93 Mutation in Neurodevelopmental Disorders
Authors: Yang Eddy , Scott Daniel , Nithianantharajah Jess 
Published: April 2020

Neurodevelopmental disorders (NDDs) such as autism spectrum disorder and intellectual disability involve aberrations in normal brain development. Human genomic studies of NDDs continue to identify mutations that converge on genes encoding postsynaptic proteins, including the synaptic scaffold DLG2 (PSD‐93 protein). Currently, we still do not fully understand the functional significance of DLG2 mutations and how the protein is impacted in these NDDs. To shed light on this, we investigated a novel DLG2 variant that has been identified in two brothers with intellectual disability and autistic‐features. We found the mouse Dlg2 R714N mutation (corresponding to human R837N) did not greatly change expression levels of PSD‐93 in HEK293 cells. We also found that binding of R714N PSD‐93 to a known interacting partner protein was largely unaffected. However, structural analyses showed that the R714N mutation may alter the conformation of the protein which results in decreased protein stability when protein‐bound. Therefore, our results show that the human DLG2 R837N missense variant likely leads to subtle changes in the shape and stability of the PSD‐93 protein. This may impact its structural role to dynamically interact and reorganise proteins at the postsynapse and thus, may impact synaptic remodelling and plasticity. Subsequent work is underway to assess this mutation in mouse neuronal cultures by examining impacts to synapse number and signalling. This work will help elucidate mechanisms of DLG2/PSD‐93 dysfunction and will increase our understanding of the functional impact of postsynaptic gene mutations in NDDs.Support or Funding InformationResearch supported by the Australian Government Research Training Program Scholarship
The localization and topography of the arteries on the middle forehead region for eluding complications following forehead augmentation: Conventional cadaveric dissection and ultrasonography investigation
Authors: Phumyoo Thirawass , Jiirasutat Nuttapatch , Jitaree Benrita , Rungsawang Chalermquan , Tansatit Tanvaa 
Published: April 2020

Forehead augmentation with filler injection is one of the most dangerous procedures associated with iatrogenic intravascular injection resulting in the severe complications. Nonetheless, few studies have determined the explicit arterial localization and topography related to the facial soft tissues and landmarks. Therefore, this study aimed to determine an arterial distribution and topography on the middle forehead region correlated with facial landmarks to grant an appropriate guideline for enhancing the safety injection. The research procedures of this study were approved by the Institutional Review Board of the Faculty of Medicine, Chulalongkorn University (IRB No. 786/61, COA No. 149/2019) and conducted in accordance with the Declaration of Helsinki of the World Medical Association (WMA). Cadaveric dissection and ultrasonographic investigation in healthy volunteers were employed to study the forehead region. The cadavers were legally donated for medical education and research, and the participants have signed the informed consent prior to the ultrasound operation. Nineteen Thai embalmed cadavers were discovered with conventional dissection and 14 Thai healthy volunteers were investigated with ultrasonographic examination on the middle forehead. At the level of mid‐frontal depression point, the transverse distance from the medial canthal vertical line to the supraorbital artery were 9.1 mm (superficial branch) and 15.1 mm (deep branch). The depths from skin of the artery were 4.1 mm (superficial branch), and 4.3 mm (deep branch). Furthermore, the frontal branch of superficial temporal artery was detectable in 42.1% as an artery entering the forehead area. At the level of lateral canthal vertical line, the vertical distance of frontal branch was 31.6 mm, and the depth from skin of the artery was 2.7 mm. In conclusion, a proper injection technique could be performed based on an intensive arterial distribution and topography, and ultrasonographic examination prior to the injection is also suggested in order to restrict the opportunity of severe complications.Support or Funding InformationThis presentation was supported by Overseas academic presentation scholarship from Faculty of Medicine Vajira Hospital, Navamindradhiraj University.
Investigating Receptor Expression in Response to Wnt5a Isoforms in Urothelial Carcinoma Cell Lines
Authors: Hemmelgarn Jessica , Coschigano Karen T. , Malgor Ramiro 
Published: April 2020

Urothelial carcinoma (UC), the most common form of bladder cancer, has a high rate of recurrence and a low five‐year survival rate for high grade and metastatic tumors. Expression of Wnt5a, a protein that plays a role in many critical processes in cells and is involved in many different molecular signaling pathways, has been shown to correlate with UC development and pathological stage. Recently the Wnt5a gene has been shown to encode two different isoforms (Wnt5a‐L and Wnt5a‐S). Correlation of isoform expression with tumor cell phenotype suggests opposing roles, either as tumor‐promoting or tumor‐suppressing, depending on the cancer type. The effects of each Wnt5a isoform in UC, however, are not well understood.In order to understand how the Wnt5a isoforms function in the development of UC, it is important to understand which signaling pathways are being activated or inhibited by each isoform. This project aims to identify Wnt5a receptors that are differentially upregulated or downregulated in response to treatment with either Wnt5a‐L or Wnt5a‐S in several different UC cell lines derived from different stages of tumors, as well as in a primary human bladder cell line. The project will include identification and characterization of the Wnt5a receptors normally expressed in the untreated, basal state, information which is currently unknown. Wnt5a isoform treatment will be conducted using conditioned media collected from Chinese Hamster Ovary (CHO) cells that have been genetically modified to express either Wnt5a‐L or Wnt5a‐S. Using western blot analysis, we have already quantified the amount of each isoform present in the conditioned media so that equal amounts of each isoform can be used for cell treatments. We have also used western blot analysis to demonstrate that we can isolate membrane proteins from cell lysates. We will now be treating the cell lines with equal concentrations of Wnt5a‐L, Wnt5a‐S or commercially available Wnt5a as a positive control; isolating the membrane proteins from each cell line after treatment; and applying the isolated membrane proteins to custom antibody arrays in order to examine the expression of 17 different Wnt5a receptors. Results from these experiments will show what receptors are upregulated or downregulated in response to each Wnt5a isoform, which will help elucidate the signaling pathways regulated by each isoform in the pathogenesis of UC.Support or Funding InformationThis project was supported in part by funding from the Honors Tutorial College, Student Enhancement Award, and Heritage College of Osteopathic Medicine Research and Scholarly Activities Committee, all at Ohio University. Special thanks is given to Dr. Karl Willert, who provided the CHO cell lines expressing each Wnt5a isoform.
Effects of Various Concentrations of Trimethylamine‐N‐oxide on Serum and Cecal Metabolites in Female SD Rats Fed a High‐Fat Diet
Authors: Lim Taehwan , Lee Kiuk , Kim Ryun-Hee , Hwang Keum Taek 
Published: April 2020

Trimethylamine‐N‐oxide (TMAO) has been shown to be a putative promoter of cardiovascular disease. Trimethylamine (TMA) precursors, such as choline and L‐carnitine, in foods can be metabolized to TMA by some gut microbiota and further converted to TMAO in liver. Recently, gut microbiota has been found to play an important role in nutrient metabolism. It can metabolize not only nutrients but also bile acids, which could affect gut liver axis and modulate metabolism. However, the effects of various concentrations of TMAO on microbial metabolites including TMA, TMAO, individual bile acids, and short chain fatty acids in rats have been little studied. The aim of this study was to evaluate effects of dietary TMAO on levels of metabolites in serum (TMA, TMAO, and bile acids) and cecum (TMA, TMAO, bile acids, and short chain fatty acids) of female SD rats fed a high‐fat (HF) diet. Sixteen female SD rats were fed HF diet (AIN‐93G diet providing 45% of energy from fat) with or without various concentrations of TMAO (0.05%, 0.15%, and 0.3%; T0.05, T0.15, and T0.3, respectively) (w/w) in autoclaved tap water for 8 weeks. Levels of TMA, TMAO, and bile acids were determined by UPLC‐Synapt G2‐Si. Short chain fatty acid profile was determined by GC‐FID. Serum TMA and TMAO levels were significantly (p<0.05) higher in the T0.3 than control group (CON), while cecal TMA level showed the opposite response. Glycocholic and tauro‐alpha‐muricholic acids were significantly (p<0.05) higher in the serum of the T0.3 than that of the CON. Taurocholic and taurodeoxycholic acids were significantly (p<0.05) the highest in the cecum of the CON. Valeric and isovaleric acids were significantly (p<0.05) higher in the cecum of the T0.05 than that of the CON.Support or Funding InformationThis research has been supported by National Research Foundation of Korea (NRF‐2017R1D1A1B03028407) funded by the Ministry of Education of Korea.
Chronic Atrial Overload Contributes to the Slow Decay of Intramural Fibrosis in the Human Atrium: Assessment by High‐Resolution 3D Histology
Authors: Ravelli Flavia , Masè Michela , Cristoforetti Alessandro , Avogaro Laura , D’Amato Elvira , Tessarolo Francesco , Piccoli Federico , Graffigna Angelo 
Published: April 2020

BackgroundThe intramural distribution of fibrosis is considered one of the most important factors in the formation of a substrate for atrial fibrillation. Chronic atrial stretch is known to play a mechanistic role in this structural remodeling process. However, the distribution of fibrosis through the entire atrial wall is poorly characterized as well as its relationship with chronic atrial stretch. Based on these premises, this study aimed at assessing the intramural profile of fibrosis in the human atrium in patients with different degrees of atrial dilatation and arrhythmic profiles by high‐resolution 3D histology.Methods and ResultsSerial vibratome‐cut longitudinal slices, encompassing the whole atrial wall at a resolution of 5 μm, were obtained from right atrial appendage samples from cardiac surgery patients. Serial slices were stained in Picrosirius red, imaged by polarized light microscopy, and analyzed with a custom‐made computer program. In all patients the intramural fibrotic content followed a progressive decreasing profile along tissue depth, passing from the epicardium to the endocardium. On average it decreased from 69 ± 12 % to 34 ± 16 % within the first 20% of atrial tissue depth, undergoing a further slight decrease to 21 ± 14 % at 40% of tissue depth. Finally, in deeper mid‐wall to subendocardial layers, the average fibrotic content slightly varied around low values of 10–13%. Distinct intramural fibrotic profiles were observed in sinus rhythm patients with normal atria with respect to patients with dilated atria, who showed, respectively, rapid and slow exponential decays (decay constant: 80.9 ± 24.4 μm versus 171.2 ± 54.5 μm, p<0.005). Similar to patients with atrial dilatation, patients with atrial fibrillation showed a slow fibrotic decrease with decay constant of 142.9 ± 41.7 μm. The decay rate of intramural fibrosis correlated with the degree of atrial dilatation (r=0.69, p<0.001).ConclusionsThe quantification of the intramural profile of fibrosis in the human atrium is feasible by combining slice‐to‐slice histology with computerized image analysis. Our results showed a deeper penetration of fibrosis in the atrial wall closely related with the presence of atrial dilatation. The stretch‐related intramural penetration of fibrosis may contribute to epiendocardial dissociation of electrical activity and the formation of an arrhythmic substrate.Support or Funding InformationThis work was supported by Fondazione Cassa di Risparmio di Trento e Rovereto.