bluuebunny/crossref_metadata_2025_split huggingface dataset:
Is the Shape of the Proximal Femur a Risk Factor for Degenaritve Processes of the Hip and Hip Fracture? A New Approach Based On Geometric Morphometrics Methods
Authors: May Hila , Rühli Frank 
Published: June 2021

The aim of the study is to establish a valid, reliable and sensitive geometric morphometrics (GM) model of the proximal femur that will allow examining the association between proximal femur shape and degenerative processes at the hip joint and hip fracture.The study included CT scans of 6 chimpanzees and 50 humans, normal femora. The surface of femora was reconstructed using Amira®. A dedicated GM protocol was developed for this purpose and includes positioning and analysis of 139 landmarks and semilandmarks using EVAN Toolbox. Reliability of the protocol was examined by repeating the measurements blindly 3 times on 5 individuals. Its validity was tested by comparing human and chimpanzees' femoral 3D model and between right and left femora within the same individual. Sensitivity of the protocol was examined by its ability to differentiate between different human populations.The protocol's reliability, validity and sensitivity showed good results. The mean Euclidean distance between individuals (0.12) was greater than within individuals (0.05). The GM protocol differentiates between human and chimpanzee and between different human populations, yet no difference in variability was found between right and left femora.The new methodology will enable us to examine shape variation in the proximal femur and reveal its association with hip maladies. This may be useful in preventive actions and treatment decisions.
Deltoid Muscle: Patterns of Innervation from Axillary Nerve
Authors: Lima Larissa , Itezerote Ana , Hojaij Flávio , Andrade Mauro , Jacomo Alfredo , Akamatsu Flávia 
Published: June 2021

IntroductionThe injury of the axillary nerve has been reported and it shows weakness of the deltoid muscle, mainly after intramuscular injections. The wrong site of these injections can cause this complication. A sharp knowledge of the deltoid muscle's innervation can avoid this iatrogenic occurrenceStudy objectiveDetermine patterns of deltoid muscle's innervation (from the axillary nerve) in order to establish a danger area (to be avoided during injection procedures)Material and MethodsTwo dissections were performed stratigraphically in human cadavers.1. Longitudinal incision in the proximal region of the arm ( anterior)2. The muscle was all exposed, their insertions were divided, and its deep part was exposed.3. This maneuver permitted to find the axillary nerve and its branchesTaking the view of the nerves, we made projections of the entry points in the muscle using a Cartesian coordinate system (the referential point was the greater tubercle of the humerus). The group of these points, when projected in the coordinated system, defined an area where, in the superficial part of the muscle, we can predict the entry of the branches of the nerve in the muscle.Summary of ResultsPreliminary results show that nerve entry points are distributed predominantly in the middle region of the deltoid muscle across the longitudinal axis of it. This area is an ellipsoid shape.ConclusionOur data can help avoiding iatrogenic injuries during procedures and, they also help understanding the myofascial pain and its treatment.
Immunohistochemical Protocol to Identify Glial Fibrillary Acid Protein (GFAP) in the Dorsal Horn of the Spinal Cord
Authors: Silva Greice Anne , Sanada Luciana , Machado Nathalia Leilane , Carmo Elisabete , Oliveira Alexandre , Fazan Valeria Paula 
Published: June 2021

Pain in neonates can induce long‐term changes in areas responsible for processing nociceptive stimulus [1]. Astrocytes are active in supporting and controlling neuronal signaling and in response to noxious stimuli they may undergo morphological changes, such as hypertrophy and increased expression of glial fibrillary acid protein (GFAP). We showed that rats receiving painful stimuli in the neonatal period presented changes in sensory, motor and limbic cortical areas related to nociception even after 180 days after the stimulus [1]. Alterations on the dorsal horn of the spinal cord in this model of neonatal pain were not yet investigated and to do so, we are testing a GFAP immunostaining protocol. Four male Wistar rats at 15 and 180 days of age were anesthetized and perfused with 4% buffered paraformaldehyde. Transverse sections of the spinal cord were cut at L4‐L6 levels and frozen. Cryosections (20 µm) were incubated in primary antibody polyclonal rabbit anti‐GFAP for 4 hours (1:1.500) followed by polyclonal donkey anti‐rabbit IgG for 45 minutes (1:250). Slides were washed in PBST, mounted in a mixture of glycerol/PBS (3:1) and observed with a light microscope (TS‐100, Nikon, Tokyo, Japan) equipped with a high resolution camera. Our results show that the method is reliable and reproducible for GFAP identification and quantification in the spinal cord of normal rats. Future studies will use this method to better understand the spinal cord response to noxious stimulus in the neonatal period. [1] Sanada et al 2014, Int J Dev Neurosci 35,55Support: FAPESP, CNPq, CAPES
ProBDNF Negatively Stimulates the Neuronal Precursor Cells Differentiate into Mature Neurons in Rat DRG
Authors: wei Ma , jinwei Yang , jin Ru , tao Luo , dongyan Wang , xingtong Li , tongtong Wang , liyan Li , jianhui Guo 
Published: June 2021

Brain‐derived neurotrophic factor precusor (proBDNF) is present in many regions of the central nerve system (CNS) and is released in cultured cortical neurons and at hippocampal aynapses. ProBDNF can induce cell apoptosis and has inhibitory effects on neuronal events, its action mode and pathway different from mature BDNF, which lead to the survival effects. Most of the research carried out on proBDNF has concentrated on the brain, cerebellum, cortex, and hippocampus.However, the roles of proBDNF in peripheral nervous system (PNS) have not yet been reported. Our research successfully separated and purified the satellite glial cells (SGCs) from normal rat dorsal root ganglion (DRG), and labelled neurons and glial cells using Immunofluorescence. GS+, GFAP+ and Tuj1‐ cells were detected in control groups, but GS+ and Tuj1+ cells were found in groups induced by proBDNF antibody. We conclude that proBDNF may participate in the regulatory process of neuronal precursor cells differentiated into mature neurons in rat DRG.*Corresponding authors: Li‐yan Li (kmliyanl@163.com) and Jian‐hui Guo (guojianhuikm@163.com)
Effects of Acute Insulin Treatment in the Recurrent Laryngeal Nerve Myelinated Fiber Morphometry in Experimental Diabetes
Authors: Mendes Vânia Alice , Silva Greice Anne , Castania Jaci , Salgado Helio , Fazan Valéria Paula 
Published: June 2021

Reports on vocal cord palsy in diabetic patients are increasing in the literature but few reports deal with the morphology of the RLN in experimental models of diabetes. Also, effects of acute insulin treatment in experimental diabetes are also rarely reported. In the present study, male Wistar rats received a single intravenous streptozotocin (STZ) injection in order to induce diabetes (N=7) while control rats received vehicle (N=9). STZ + insulin animals (N=8) received a daily subcutaneous insulin injection three days after STZ injection. On the 15th day after STZ or vehicle injections, rats were anesthetized for arterial pressure (AP) and heart rate (HR) recordings, prior to RLN removal. Proximal and distal segments of the right and left RLN were prepared for light microscopy morphometry, carried out with the aid of a computer software. Myelinated fiber morphometry showed a tendency (P = 0.07) towards a significant difference in the fiber and axon diameter between control and STZ+insulin, tending to be larger in the control group. The opposite tendency (p=0.07) was observed for the G ratio values, being larger on STZ+insulin group compared to control. Our results show that acute treatment with insulin affects negatively the size of myelinated fibers and axons in the recurrent laryngeal nerve.Support: FAPESP, CNPq, CAPES and FAEPA
The Effects of Methylene Blue on Autophagy and Apoptosis in MRI‐defined Normal Tissue, Ischemic Penumbra and Ischemic Core
Authors: Zhou Changman , Jiang zhao , Lu Jianfei , Wang Ke , Duong Timothy Q 
Published: June 2021

Background and purposeMethylene blue (MB) USP, which has energy‐enhancing and antioxidant properties, is currently used to treat methemoglobinemia and cyanide poisoning in humans. We recently showed that MB administration reduces infarct volume and behavioral deficits in rat models of ischemic stroke and traumatic brain injury. This study reportsthe underlying molecular mechanisms of MB neuroprotection following transient ischemic stroke in rats.MethodsRats were subjected to transient ischemic stroke. Multimodal MRI during the acute phaseand at 24hrs was used to define three regions of interest (ROIs): i) the perfusion‐diffusion mismatch salvaged by reperfusion, ii) the perfusion‐diffusion mismatch not salvaged by reperfusion, and iii) the ischemic core. These ROIswere extracted for western blot analyses of autophagic and apoptotic markers.ResultsThe major findings were: 1) MB treatment reduced infarct volume and behavioral deficits, 2) MB improved CBF to perfusion‐diffusion mismatch tissue after reperfusion and minimized harmful hyperperfusion24hrs after stroke, 3) MB inhibited apoptosis and enhanced autophagyin the perfusion‐diffusion mismatch tissue, 4) MB inhibited apoptotic signaling cascades (p53‐Bax‐Bcl2‐Caspase3), and 5) MB enhanced autophagic signaling cascades (p53‐AMPK‐TSC2‐mTOR).ConclusionsMB induced neuroprotection,at least in part,by enhancing autophagy and reducing apoptosis in perfusion‐diffusion mismatch tissue following ischemic stroke.
Characterization of Neural Stem/Progenitor Cells in the Brain of the Leopard Gecko (Eublepharis macularius)
Authors: Macdonald Alaina , McDonald Rebecca , Gilbert Emily , Vickaryous Matthew 
Published: June 2021

Many lizard species can regenerate their spinal cord following tail loss. There is increasing evidence to suggest that populations of neural stem/progenitor cells (NSPCs) in the original spinal cord are activated after the tail has been lost, and contribute to regeneration. For the brain, less known. Here, we used a panel of NSPC markers and a bromodeoxyuridine (BrdU) pulse‐chase experiment to characterize the presence of potential NSPCs in the adult brain of the leopard gecko. Prior to tail loss, we determined that some cells bordering the ventricular system (including the lateral, third and fourth ventricles) are slow‐cycling (retained BrdU for 20 weeks) and express several widely recognized NSPC markers including Sox2, Sox9 and GFAP. Following tail loss, subsets of these same cells begin to proliferate (as evidenced by PCNA immunostaining) and express an additional NSPC marker, Musashi‐1. These findings offer compelling support for the presence of NSPCs in the adult lizard brain, and suggest that similar to the spinal cord, these populations are activated in response to injury.NSERC Grant:400358
Beneficial Effect of (‐)‐epigallocatechin‐3‐gallate (EGCG) on Sciatic Nerve Crush Injury: Stereological analysis
Authors: Renno Waleed , Khan Khalid , Benov Ludmil 
Published: June 2021

The neuroprotective effect of EGCG on sciatic nerve crush injury is associated with significant neurobehavioral recovery (Renno et al. 2014). In this study, we examined the histological changes in the sciatic nerve after crush injury following EGCG treatment. The mean cross sectional area of the axon showed a significant increase in the EGCG treated group starting at week 4 and continued up to week 8 compared to CRUSH animals. However, axon area remained much smaller in CRUSH+EGCG and CRUSH groups compared to SHAM and NAIVE animals. The number of axons per unit area was significantly decreased in CRUSH+EGCG and CRUSH groups compared to control. The CRUSH group displayed a significant increase in total small and thinly myelinated axon number at week 6 and 8 post‐injury. ECGC treated animals showed a significant decrease in the total myelinated axon numbers. CRUSH+EGCG group showed a significant increase in myelin thickness in the distal sciatic nerve segment at week 4 till week 8 compared to CRUSH animals. EGCG treatment significantly increased the fiber diameter to axon diameter (F/A) ratio and the myelin thickness to axon diameter (M/A) ratio starting at week 2 post sciatic nerve injury compared to CRUSH animals. EGCG has profound positive effect on the myelinated axons; increase in the axon area, myelin thickness, F/A ratio and M/A ratio. In summary, quantitative stereological evaluation revealed significant histomorphological evidence of neuroregeneration in the sciatic nerve of CRUSH+EGCG rats compared to control groups between 2 and 4 weeks postoperatively. The morphometric data analysis supports our hypothesis that EGCG treatment stimulates regeneration of myelinated axons and minimizes histomorphological alterations caused by crush injury of sciatic nerve.This work was supported by Kuwait University Research Administration (grant No. MA01/11).
Modulation of MAP2 and Tau Following Alpha Lipoic Acid Supplementation Ameliorates Arsenic Induced Adverse Effects in Developing Rat Cerebellum
Authors: Dhar Pushpa , Kaushal Parul , Mehra Raj 
Published: June 2021

The present study was designed to evaluate the effects of ALA on arsenic (iAs) induced alterations in developing rat cerebellum. Pregnant Wistar rats, procured from Experimental Animal Facility after obtaining ethical clearance from Institutional Ethical Committee, were maintained on a 12 hr light‐dark cycle with ad libitum access to food and water. The day of birth of pups was designated as postnatal day (PND) 0. Mother reared pups were divided into control and experimental groups, the later receiving either NaAsO2 alone or along with ALA from PND 1 to 21. Behavioral parameters (cliff avoidance, tail hung, negative geotaxis, Rota‐rod) were carried out at different time points. Animals were sacrificed on PND 22 and the fresh cerebella were processed for estimation of levels of iAs, malondialdehyde (MDA) and glutathione (GSH). Cerebella from perfusion fixed animals were processed for cryo‐sectioning and immunohistochemical localization of Microtubule associated protein (MAP2) and Tau. The observations of present study revealed behavioral deficits in pups exposed to NaAsO2 alone. Co‐administration of ALA with iAs resulted in improved behavioral parameters, decrease in iAs and MDA levels, increase in GSH levels along with upregulation of MAP2 and Tau expression suggesting thereby protective role of ALA in NaAsO2 induced neurotoxicity.DST INSPIRE Fellowship, Govt. of India (110053) to Parul Kaushal is acknowledged.
Purification of DNA From Chicken Embryos to Characterize Genetic Sequences
Authors: Kadiyala Usha , Smith Sheila , Benore Marilee 
Published: June 2021

Riboflavin (Rf), Vitamin B2, is one of several antioxidant B vitamins that organisms need for survival. It is significant in fat, protein and energy metabolism, and plays a key role in maintenance of the nervous system and vision. In oviparous species, Riboflavin Binding Protein (RBP) transports Rf from the bloodstream of the laying hen into the eggs, where the vitamin is essential for embryonic development.Our research group maintains a flock of natural mutants, white leghorn chickens that contain s splice‐site mutation in the the gene for RBP. The Rf deficient egg embryos can be rescued by addition of supplemental Rf. In order to further characterize gene and protein structure and function, we have developed a modified assay to purify the DNA from chick embryos. Using kits and modification of the procedures, including flash freezing extracted embryos and altering the lysis period of the tissue, resulted in better DNA yield and purity. This embryonic DNA can be used for further characterization of the RBP and other genes.
Changes in Water Availability or Molecular Crowding Induce Folding in Instrinsically Disordered Stress Proteins from Plants
Authors: Covarrubias Alejandra , Cuevas‐Velazquez César , Rivera‐Najera Lucero , Rendon‐Luna David , Saab‐Rincón Gloria , Martinez‐Yamout Maria , Dyson H Jane , Amero Carlos 
Published: June 2021

Late Embryogenesis Abundant (LEA) proteins are a broadly distributed group involved in the plant response to water deficit. Most of them are predicted to be intrinsically disordered. By partial dehydration and freeze‐thaw in vitro assays, it was shown that some LEA proteins protect other proteins from the effects of water deficit. Given the presumed unstructured character of LEA proteins, we hypothesized that their flexible nature plays a critical role in their interaction with diverse partners. This structural flexibility might be modulated by the cell water status, promoting conformations needed to interact with specific targets. In this work, we report some of the structural and functional properties of LEA proteins from groups 4 and 6. We showed that proteins in both groups are intrinsically unstructured in solution; however, their intrinsic potential to form secondary structure was exhibited by treatment with a structure inducer, which promoted a significant α‐helix formation. A decrease in water availability and an increase in molecular crowding induced folding in both LEA protein groups but in different level. We also determined the protection efficiency of group 4 LEA proteins and proposed a mechanism for this function.This work was partially supported by CONACyT (132258) and DGAPA‐UNAM (IN208212).
Total Mutagenesis and the Effects on the MDM2‐p53 Interaction
Authors: Beyer George , Thayer Kelly 
Published: June 2021

The aim of this study is to assess the impacts of mutation on the p53‐MDMD2 interaction.While the available literature explores alanine‐scanning mutagenesis of the six necessary amino acids of the p53 binding subdomain in order to obtain precise ΔG values for the wild‐type amino acids in the p53‐MDM2 binding domain, the effects of non‐alanine point mutations have not been explored. Many mutations of MDM2 that could affect the binding to p53 have largely gone unstudied, but several SNPs in MDM2 have been directly linked to tumor formation. The aim of this study is to assess the impacts of mutation on the p53‐MDMD2 interaction utilizing both MUMBO and MM‐PBSA. MUMBO was used in order to perform total mutagenesis of the p53‐MDM2 domain in order to determine key mutations that may influence MDM2 binding activity. Data achieved from exhaustive point‐by‐point mutagenesis suggests great variation in the mutant structures. Mutants of interest will then be further explored utilizing the MM‐PBSA approach, and structural models will be constructed. Many mutations not shown through alanine‐scanning mutagenesis may greatly impact the p53‐MDM2 interaction.
Intrinsic Structural Disorder Observed in the Cell Cycle Regulatory GTPase Nucleostemin
Authors: Daman Tyler , Robinson Victoria 
Published: June 2021

To maintain tissue homeostasis, it is necessary to control cell proliferation based on nutrient availability and environmental factors. Numerous lines of evidence suggest that the balance of the cellular concentrations of nucleostemin (NS), a non‐canonical circularly permuted GTPase, is critical for cell cycle progression. It does so by interacting with different partners involved in a wide range of cellular processes such as ribosome biogenesis, tumor suppression, and DNA maintenance. The cellular function of NS is regulated by guanine nucleotide driven localization to the nucleolus. Currently, little is known about the structure of NS and how it is modulated during this event.Using an ensemble of biophysical techniques, we present the first structural characterization of Drosophila melanogaster Nucleostemin (dNS1), which is 33% identical and 67% similar to the human homolog. This represents the first structural data collected for a eukaryotic cpGTPase. Bioinformatic analysis, limited proteolysis, and circular dichroism were used in combination to determine that dNS1 contains large intrinsically disordered regions, primarily at the N‐ and C‐ termini. Small‐angle X‐ray scattering (SAXS) experiments, supported by analytical ultracentrifugation, provided a solution description of dNS1. Data obtained from these techniques establish that dNS1 exists as a highly elongated monomer, and confirms the lack of secondary structure. In summary, our studies show that NS contains large regions of structural disorder, which likely facilitates its broad partner recognition.
The Intrinsically Disordered C‐terminal Domain Triggers Nucleolus Localization and Protein Interactions of Poly(A)‐specific Ribonuclease
Authors: Yan Yong‐Bin , He Guang‐Jun 
Published: June 2021

Poly(A)‐specific ribonuclease (PARN) is a highly regulated deadenylase with diverse physiological functions via modulating poly(A)‐tail length of eukaryotic mRNAs. PARN is a multi‐domain protein with three well‐structured RNA‐binding domains and one C‐terminal domain (CTD) with unknown functions. Herein we investigated the structural features and potential functions of CTD. Spectroscopic studies indicated that CTD was intrinsically disordered with some local structures. The full‐length PARN had an intensive distribution in the nucleoli, while truncations of CTD impaired the nucleolus location of PARN. The nuclear‐localization and nucleolus‐localization sequences were identified by both deletion studies and fusion of the sequence with GFP. Furthermore, CTD was found to be the site triggering protein‐protein interactions and could selectively bind with its partners via the status of phosphorylation. Thus the intrinsically unstructured CTD is indispensible in the nucleolus localization and binding with PARN partners, which in turn regulate PARN activity. We proposed a model in which PARN is resistant in the nucleoli and can recruit dissimilar binding partners via the intrinsically disordered CTD to respond to various cellular stresses.This study was supported by grant 31370797 from the National Natural Science Foundation of China.
Specificity of the Yeast Auxilin Homolog Swa2 in [URE3] Prion Propagation is Solely Determined by the TPR Domain
Authors: Hines Justin , Troisi Elizabeth , Rockman Michael , Oliver Emily 
Published: June 2021

Previous results from our laboratory assert that the yeast Hsp40 chaperone protein Swa2 is essential for the propagation of the prion [URE3] in a yeast population. Swa2 is the sole yeast homolog of mammalian protein auxilin, which functions in vesicle‐mediated endocytosis by disassembling the structural lattice formed by the protein clathrin. The objective of this study was to find the determinants of the specific genetic interaction between Swa2 and [URE3]. The primary method used in this study was complementation by plasmid‐shuffling in the budding yeast Saccharomyces cerevisiae. Summary of Results: Our investigation revealed that two regions of the Swa2 protein, the J‐domain and the tetratricopeptide repeat region (TPR) are both necessary and sufficient to maintain [URE3], but that the TPR domain is solely responsible for prion/chaperone specificity. While the function of the TPR domain in Swa2 is undetermined, TPR domains typically act as docking platforms for a variety of protein complexes. As such we propose that the Swa2 TPR domain either participates in a bipartite interaction with Hsp70, analogous to the C‐terminal domains of other yeast Hsp40s, or that the TPR domain mediates the formation of a ternary complex with another undetermined protein in [URE3] propagation. We expect that understanding the function of the Swa2 TPR domain will provide new insight into both J‐protein function and the complex mechanisms of prion propagation.This work was supported by a Single‐Investigator Cottrell College Science Award (Award #21010) from the Research Corporation for Science Advancement and by by the National Institute Of General Medical Sciences of the National Institutes of Health under Award Number R15GM110606.