bluuebunny/crossref_metadata_2025_split huggingface dataset:
Manipulation of Igf‐1 Expression in the Placenta and Impact on the Fetal Brain
Authors: Carver Annemarie , Taylor Robert , Kundu Sreelekha , Stevens Hanna 
Published: May 2022

The placenta is an essential organ involved in mammalian development in utero. The main function of the placenta is the control of the transfer of nutrients and waste between mother and fetus, and production and delivery of hormones and growth factors. There is a lack of research that investigates the role of maternal and fetal genetics and molecular biology of the placenta’s impact on fetal neurodevelopment. Igf‐1 is a gene involved in mediating fetal development as well as growth of the brain. It is also involved in regulation of placental growth and transport. The placenta is the main source of fetal Igf‐1. Igf‐1 gene expression is altered in some individuals with autism spectrum disorder (ASD) which may reflect a role of Igf‐1 in ASD etiology. We previously found that prenatal stress, a risk factor for ASD, increased placental Igf‐1 expression and increased proliferation of striatal neural progenitors which express Igfreceptors. Developmental striatal changes are found in ASD. We therefore hypothesized that placental alterations in Igf‐1 lead to ASD‐relevant brain changes.MethodsWe examined this through the use of CRISPR Igf‐1 overexpression or control plasmids directly injected in mouse placenta on embryonic day 12, with subsequent tracking of changes in placenta, fetal body, and fetal brain. Igf‐1 overexpression in placenta had a variable trajectory. While placental weight was unaffected by Igf‐1 overexpression at E18, we found developmental and morphological changes of the embryo, including larger body and larger size and altered morphology of the head (macrocephaly) at E18. Brain volume measures demonstrate a range of effects on the primordial cortex and striatum at E14 and E18. One outcome of this work is advancements in methodology for understanding how placental genetics affect the development of the fetal brain. We also conclude that placental Igf‐1 expression does influence offspring growth and ASD‐relevant brain outcomes which suggests a role for placental genetics in neurodevelopmental disorders.
Neurons in the Caudate Nucleus of the Camel and Human: A Comparative Study
Authors: Bataineh Ziad M. 
Published: May 2022

The caudate nucleus is described as a huge, subcortical grey mass in the forebrain known for its role in motor function and involved in a number of clinical syndromes. During ontogenesis, the caudate nucleus follows the curvature of the telencephalic vesicle, hence its curved shape. Its rostral portion referred to as the head, is far more voluminous than the body.AimsThis study aimed to describe the different types and characteristics of neurons in the caudate nucleus of the brain of camel and human and compare these neurons with their counterparts in other species.Method4mm thick blocks containing caudate nucleus was cut, dry and processed according to the rapid Golgi and or the Golgi‐Kopsch methods (double impregnation). Well‐impregnated neurons studied under light microscope.ResultsAccording to the present study, neurons were classified according to the density and distribution of the spines along their axons and dendrites. Neurons in this study classified into three main types in the caudate nucleus of camel and human. According to the somal size each group includes one or more of the following cell size: large, medium and small. Rich‐spiny neurons (Type I): include medium and small subtypes. Hairy‐like, mushroom‐like and stubby spines were present in the secondary dendrite and their branching. Sparsely‐spiny neurons (Type II): they have relatively larger cell bodies and thicker primary dendrites than (Type I). Characterized by their very low density of spines and poorer branching of their dendrites. Aspiny‐neurons (Type III): are large, medium and small size. May have varicose, appendages with various types and short axon which gives collaterals mostly within the dendritic domain.ConclusionThe neuronal structure of the caudate nucleus in the camel is differ from that of the human. The human neurons appear slightly larger than those of the camel and they have more dendritic trunks that break up more often than the neurons of the camel. Although the types of spines are similar in both species, the density of spines appears to be greater in the camel than in the human. The structural differences of the caudate neurons between two species may result from the fact that the homo sapeneis belongs to a phylogenetically younger order (Anthropoidea) than the camel. (Artiodactyla) which support the fact that the number and diversity of the types of neurons go to increase throughout phylogenesis.
Nitric Oxide Inhibits TRPV4‐mediated Hemichannel Opening in Porcine Nonpigmented Ciliary Epithelium
Authors: Shahidullah Mohammad , Delamere Nicholas 
Published: May 2022

Previous studies showed the presence of connexin 43 (Cx43), connexin 50 (Cx50), and pannexin 1 (Panx1) in the nonpigmented ciliary epithelium (NPE) of the eye, as well as evidence of hemichannel (HC) activity. In the lens, mechanosensitive Transient Receptor Potential Vanilloid 4 (TRPV4)‐mediated calcium entry stimulates hemichannel activity. The aim here is to test for TRPV4 expression in the NPE, determine whether TRPV4 activation causes HC opening, and study the influence of nitric oxide.Immunohistochemistry was used to probe for TRPV4 in porcine ciliary body. Propidium iodide (PI) uptake and ATP release by primary cultured NPE was used to assess HC opening. PI and ATP are large molecules (MW 668 and 507, respectively) that pass through open hemichannels but not the plasma membrane. ATP in the bathing medium was measured by luciferase luminometry and PI uptake by fluorimetry.Abundant expression of TRPV4 was detected in the NPE cell layer of the ciliary body. Cultured NPE cells displayed an increase in PI uptake when exposed to a TRPV4 agonist GSK 1016790A (10 nM) or to hypoosmotic solution (200 mOsm) (Control 4.65 ± 0.24 vs GSK 10.26 ± 0.32 and Hypo 8.26 ± 0.42 fluorescence units per mg protein; n=6, p<0.001). PI uptake also was increased in NPE cells cultured on a flexible membrane and subjected to cyclic stretch for 1‐10 min (10%, 0.5 Hz), the peak response was at at 2 min (Control 1.5 ± 0.03 vs stretch 2.5 ± 0.10; n=6, p<0.001). The increase in PI uptake caused by hypoosmotic solution or by cyclic stretch was abolished by a selective TRPV4 antagonist HC 067047 (10 μM). Hyposmotic solution increased ATP release from 1.12 ± 0.24 to 2.81± 0.27 (n=12, p<0.001) while cyclic stretch increased ATP release from 0.20 ± 0.03 to 0.40 ± 0.03 pmoles/mg protein (n=6, p<0.001). ATP release responses caused by hypoosmotic solution and cyclic stretch were inhibited by HC 067047. Hypoosmotic solution‐induced ATP release was also inhibited by connexin inhibitory peptide, Gap‐27 (200 μM) and nitric oxide (NO) donors, sodium nitroprusside (SNP) and S‐nitroso‐N‐acetylpenicillamine (SNAP).TRPV4 channels are expressed by the NPE. The ability of HC 067047 and Gap‐27 to prevent ATP release and PI uptake points to TRPV4‐dependent opening of connexin HCs in NPE cells. Suppression of ATP release by SNP and SNAP indicates the hemichannel opening mechanism is inhibited by NO.
Monomeric CCTδ Interacts with Dynactin Associated Protein (dynAP)
Authors: Iwuagwu Benedicta U. , Grantham Julie 
Published: May 2022

Introduction‐The chaperonin containing tailless complex polypeptide 1 (CCT) is a molecular chaperone consisting of eight distinct protein subunits. In the monomeric form, several CCT subunits possess additional functions distinct from their oligomeric function [1]. Over‐expression of CCTδ, but not the other seven CCT subunits, results in the appearance of numerous protrusions at the cell surface termed a “protrusion phenotype” which is abrogated by mutations in CCTδ apical domain (G357D) and the ATP binding pocket (D104E) [2]. In addition, it is dependent on CCTδ binding the p150Glued of the dynactin complex. Depletion of either p150Glued or the dynactin complex‐associated transmembrane protein, dynAP prevents GFP‐CCTδ localization to the plasma membrane and blocks protrusion phenotype formation [3]. The work of others [4] shows dynAP binds p150Glued. However, biochemical evidence of CCTδ interacting with dynAP has not been shown. We hypothesized either a direct transient interaction between CCTδ and dynAP or a trimer with p150Glued occurring.Materials & Method‐Mouse melanoma B16F1 cells were cultured as described in [2], transiently transfected with CCTδ constructs, and mutants thereof fused to GFP as described in [2], and full‐length human; p150Glued with C‐terminal Myc and dynAP with an N‐terminal FLAG epitope tags as described in [3]. To stabilize transient interactions in‐vivo, confluent cells were crosslinked with 0.5 mM 3, 3′‐Dithiodipropionic acid di (N‐hydroxysuccinimide ester). Post‐nuclear supernatants from lysed cells were applied to GFP‐trap beads (Chromotek), followed by Western Blotting to detect precipitating proteins.Results‐Following the double transfection of B16 cells with Flag‐dynAP and wild type (WT) GFP‐CCTδ, there was a significant interaction between dynAP and WT CCTδ versus control, D104E. In addition, following the triple transfection of GFP‐CCTδ/p150Glued‐Myc/Flag‐dynAP, there was a significant interaction between dynAP and WT CCTδ versus control, D104E. Furthermore, following the double transfection of WT CCTδ apical domain of (AD) and Flag‐dynAP, there was a significant interaction between dynAP and WT AD versus control, G357D.Conclusion‐Here, we show for the first time an interaction between CCTδ and dynAP via immunoprecipitation representing either a direct interaction or as part of a trimeric complex with p150Glued. This provides biochemical evidence for the mechanism underlying the formation of the protrusion phenotype. This work lends more information on monomeric CCTδ function in cells and would contribute to more understanding of CCTδ’ s role in basic cell and cancer biology. More research is needed to understand the molecular effects of these interactions in cells and in‐vivo.References

Svanstrom & Grantham (2015) Cell Stress and Chaperones; 21:55–62.
Spiess, M. et al (2015) Journal of Molecular Biology; 427:2757‐2764.
Echbarthi M. et al (2018) Experimental Cell Research; 370:137‐149.
Kunoh T. et al (2010) Molecular Cancer Therapeutics; 9:2934‐2942.
Creation of Three‐Dimensional Pedagogical Models of Genetic Mutations
Authors: Paliwal Vipin , Cohan Megan 
Published: May 2022

A genetic mutation is a change of one or more bases in the DNA sequence that may alter the structure and ability of the resulting protein product to function properly. In this study, we created three‐dimensional (3D) pedagogical models for teaching genetic mutations. SolidWorks©, 3D modeling software was used to create the models showing the three main types of DNA mutations: (i) point mutation, (ii) frameshift mutation, and (iii) insertion mutation. The models were 3D printed using a selective laser sintering (SLS) machine. Further, the printed models are strong enough to absorb physical mishandling by the users such as accidental drops. User instructions accompanying the models were developed.The models were created to allow the individual nucleotide bases adenine (A), thymine (T), guanine (G), cytosine (C) and uracil (U) to be attached or removed from the DNA and mRNA backbone and moved around easily by using magnets embedded into the individual pieces. Other ease of use features include color coding and labeling emphasize the difference between the four nucleotides in DNA. Further, as an example of real‐life mutations, we also created a unique model explaining the formation of cyclobutane thymine‐thymine (T‐T) dimers following exposure of DNA to ultraviolet B (UV‐B) radiations. UV‐B radiation in the region of 290‐320 nm is absorbed by skin cells resulting in DNA damage and mutated DNA which can lead to skin cancer. The 3D model showing cyclobutane dimers should help the students gain a deeper understanding of DNA damage, and skin cancer at the biochemical level. Ultraviolet light can break bonds in adjacent thymine bases. New covalent bonds are formed, linking the two thymine bases, forming cyclobutane T‐T dimers. We believe that models will enhance in‐depth hands‐on learning of the genetic mutations.
Is there a relationship between in‐season injury risk and Y balance or vertical jump in elite youth soccer players?
Authors: Haraldsdottir Kristin , Baer Michael , Brickson Stacey , Watson Andrew 
Published: May 2022

PurposeInjuries are relatively common among elite youth soccer players, and time lost participating in sport due to injury can have detrimental psychological effects on players. The purpose of this study was to determine whether the Y Balance Test (YBT), or vertical jump can identify those players at risk of suffering injury.Methods41 female elite soccer players (age 12‐17) performed YBT and vertical jump testing prior to the competitive season. Time‐loss injuries during the following season were recorded. Injured and uninjured athletes were compared with respect to YBT composite scores (absolute and relative to leg length), asymmetry, and vertical jump. Logistic regressions were performed to examine the relationships between YBT, vertical jump, and injury risk.ResultsTwelve athletes (29%) suffered a time‐loss injury during the season. Athletes who suffered an in‐season injury did not differ from uninjured athletes with respect to age (14.9 +/‐ 1.6 v 15.3 +/‐ 1.4 years, respectively), years of experience (9.7 +/‐ 1.7 v 10.2 +/‐ 2.4 years, p=0.26), YBT composite scores (111 +/‐ 16 v 109 +/‐ 17 cm, p=0.99), YBT composite scores relative to leg length (1.29 +/‐ 0.2 v 1.27 +/‐ 0.26, p=0.85), the difference in centimeters between composite scores of each leg (7.9+/‐8.4 v 4.6+/‐3.8, p=0.09) or vertical jump (15.3±1.6 v 15.3±2.6 in, p=0.96). In‐season injuries were not significantly associated with YBT composite scores (OR= 0.99 [95% CI = .95‐1.04] ,p= 0.80), relative YBT composite scores (1.0 [.97‐1.03], p=0.98) or vertical jump scores (0.98 [.72‐1.33], p=.91). Although not reaching statistical significance, higher asymmetry in composite scores showed some evidence of an increased risk of subsequent injury (1.2 [0.97‐1.42], p=0.10).ConclusionsThe composite YBT score and vertical jump did not predict in‐season injury incidence in this sample of elite female adolescent soccer players. Higher YBT asymmetry may be associated with an increased risk of in‐season injury, although this study may have been limited by sample size to identify a statistically significant difference.SignificanceAlthough the YBT is widely used as an injury risk screening tool, our findings suggest that it may not be a strong predictor of subsequent time‐loss injuries in elite female adolescent soccer players.AcknowledgementsWe are grateful to the adolescent soccer players who participated in the study.
A Single Glucocorticoid Injection Accelerate Erythropoiesis but Does Not Improve Performance
Authors: Bejder Jacob , Bonne Thomas C. , Breenfeldt Andersen Andreas , Sørensen Henrik , Nordsborg Nikolai B. 
Published: May 2022

The World Anti‐Doping Agency prohibit glucocorticoid (GC) administration during competition but allow administration in periods out of competition. However, GC administration may accelerate erythropoiesis and resemble blood volume manipulation methods, which are prohibited at all times. Here, we investigated whether a single GC injection accelerate erythropoiesis, increase total hemoglobin mass (Hbmass) and improve exercise performance.In a randomized, double‐blinded, placebo‐controlled crossover design (3‐month washout), eight trained males (maximal oxygen uptake 60±5 ml O2·min‐1·kg‐1) were injected with 40 mg triamcinolone acetonide (GC group) or saline (PLA group) in the gluteal muscles. Venous blood samples collected before and 7‐10h, 1, 3, 7, 14 and 21 days after treatment were analyzed for [Hb] and reticulocyte percentage (ret%). Hbmass and mean power output in a 450‐kCal time trial was measured before as well as one and three weeks after treatment.A 27±28% and 54±41% higher ret% was evident three (P<0.01) and seven (P<0.001) days after the GC administration, respectively, compared with PLA, whereas [Hb] was similar between groups. Additionally, Hbmass was 20±28g higher (P<0.05) one week after GC administration compared with PLA, but similar after three weeks. The mean power output was similar between groups one (GC: 285±70 W, PLA: 279±69 W) and three (GC: 279±72 W, PLA: 276±71 W) weeks after treatment.In conclusion, a single intramuscular injection of triamcinolone acetonide was sufficient to accelerate erythropoiesis and increase Hbmass, but did not translate into improved aerobic exercise performance in the present study. Future research should confirm whether anti‐doping authorities should consider glucocorticoid injections as a method of blood volume manipulation.
Vaping /E‐liquid Exposure Causes Dysregulation of Neutrophil Extracellular Trap formation
Authors: Abdelwahab Sabri , Ghosh Arunava , Wrennall Joe , Tarran Robert 
Published: May 2022

IntroductionElectronic cigarettes (e‐cigarettes) vaporize e‐liquids containing propylene glycol and vegetable glycerine (PG/VG), nicotine, and flavors. The inhaled aerosol from e‐liquids may be potentially toxic or harmful. Although the short and long‐term health consequences of e‐cigarettes use are poorly understood, their use has risen steeply among adolescents and young adults. Neutrophils play an essential role in innate immune and are the first responders during infections. Thus, this study aims to test whether vaping can affect neutrophil function. We hypothesized that vaping e‐cigarettes and exposure to e‐liquids activate neutrophils, cause degranulation, and induce extracellular traps (NETs) through altered calcium signaling.Method and MaterialsNeutrophils were isolated from human blood samples of non‐vapor control (n=9) and e‐cigarette users (n=5) and studied within 2 hours. To measure changes in cytosolic calcium, neutrophils were loaded with Fluo‐4. Intracellular calcium responses were monitored on a Tecan multi‐plate reader following stimulation by 1 µM thapsigargin and 1 µM fMLP/100ng per ml LPS. In addition, since elevations in intracellular calcium trigger neutrophil activation and degranulation, elastase activity in the media samples was measured using a specific fluorogenic substrate (MAA‐3133, © vivitide, LLC.) at excitation/emission wavelengths 380/460 nm. To study NET formation, neutrophils were labeled with SYTOX Green dye and then exposed to different concentrations of Juul e‐liquids (menthol, classic tobacco flavors) and Bidi Stick flavors (Arctic, Gold, and Solar). Data acquisition was performed with Cytation‐5 Cell Imaging Plate Reader, then analysis and representative images were captured by confocal microscopy.ResultsNeutrophils from e‐cigarette users (n=5 vapers) showed significantly (P < 0.001) increased intracellular calcium responses to thapsigargin and fMLP/LPS treatments compared to the non‐user controls (n=9). Additionally, vapers' neutrophils also secreted more elastase (P < 0.001) compared to the control non‐users, indicating that e‐cigarette users may be susceptible to increased systemic protease activity. Our data also indicated that Juul and Bidi Stick e‐liquids stimulated neutrophils to induce NETs in a dose‐dependent manner. Furthermore, the study revealed that Juul e‐liquid menthol flavor with 5% nicotine could significantly (P < 0.001) release NETs than equivalent nicotine ± benzoic acid alone.SummaryCirculating neutrophils are normally quiescent. However, our data indicated that circulating neutrophils from vapers were abnormally active and degranulated by released elastase. Additionally, e‐cigarette liquids dysregulated neutrophils by increasing NETs production. Overall, we speculate that vaping imbalanced innate immune system and may lead to abnormal responses to the infection, inappropriate inflammatory responses, and adverse health effects.
Extracellular Vesicles from iPSC derived Muscle Progenitor Cells Rejuvenate the Dysfunctional Muscle Stem Cells during Aging
Authors: Xuan Wanling , Tipparaju Srinivas M. , Ashraf Muhammad 
Published: May 2022

Background and ObjectiveSarcopenia is defined as the loss of skeletal muscle mass and function due to age, which is a major concern for life long health and wellbeing in elderly population. Age‐associated decline in the performance of muscle stem cells (MuSC) contributes to the deterioration of skeletal muscle during aging. Using small molecule Givinostat and CHIR99021 we previously demonstrated the generation of muscle progenitor cells (Givi‐MPC) of human Induced pluripotent stem cells (iPSC) from multiple cell lines. We found that extracellular vesicles (EVs) from these MPC have a specific cargo compared with human myoblasts. Here, we investigated whether these EVs can rejuvenate the aging dysfunctional muscle stem cells.Methods and ResultsUsing size exclusion chromatography EVs were isolated and purified from iPSC derived Givi‐MPC and primary human myoblasts. The EVs were visualized by transmission electron microscopy. Next, we isolated primary MuSC from 25 months old aged C57/B6 mice, which were confirmed by the expression of myogenic transcription factor, Pax7. We found the aged MuSC pretreated with EVs from MPC promoted clonal expansion and restored myogenic potential in vitro with more myotubes formed after 72h differentiation in medium with 2% horse serum compared with PBS pretreated MuSC (MuSC (Fusion index: 40.3±5.2% vs.15.0± 2.1%, n=3).). To test the regenerative potential of the EVs on aged MuSC, 8 weeks old mdx/scid mice (dystrophin deficient mice) were irradiated to destroy the host MuSC and then injured their tibialis anterior (TA) muscle with cardiotoxin (CTX). 24 hours after injury, we transplanted the aging MuSC (5000 cells) either with EV or PBS pretreatment into TA muscles, respectively. 5 days after transplantation, the EVs pretreated MuSC increased muscle regeneration with muscle fibers positive for embryonic myosin heavy chain compared to PBS pretreated MuSC. One month later, we found transplantation of EVs pretreated MuSC restored dystrophin expression in these mdx mice. We further tested the effects of EVs pretreatment on MuSC in 25 months old mice following CTX injury. Consistent with the results in mdx mice, pretreatment of mice with EVs from Givi‐MPC showed enhanced regeneration (6‐fold increase in number of new muscle cells with central nuclei) in CTX injured aged muscle accompanied by an increase of Pax7 positive MuSC compared with PBS treated mice or treatment with EVs from human myoblasts (number of Pax7 positive MuSC: EVs‐Givi‐MPC 10±5/mm2; PBS 2±1/mm2; EVs‐human myoblast 6±2/mm2, n=3).ConclusionThe regenerative potential of aging MuSC is restored with the treatment of EVs from Givi‐MPC allowing stem cell renewal and differentiation upon activation.