bluuebunny/crossref_metadata_2025 huggingface dataset:
Quantifying the Shape of the Greater Sciatic Notch using Elliptical Fourier Analysis
Authors: Byram Jessica , Latham Krista , Nawrocki Stephen
Published: June 2021

The greater sciatic notch of the coxa is well established as a sexually dimorphic feature in humans, with wider notches being more common in females. However, the size and shape of the notch can vary between populations, which can complicate the use of this feature as a sex determinant in forensic analyses. The current study investigates the relative contributions of variables that may influence the shape of the greater sciatic notch, including sex, ancestry, age at death, and environmental effects (secular change). As the notch is a complex curvilinear shape not easily quantified using standard linear measurements, a more sensitive morphometric approach was used. The study sample included 975 dry bone specimens obtained from three skeletal collections, including three ancestral groups from recent (1950‐present) and earlier (pre‐1950) time periods. Each coxa was photographed and the contour of the greater sciatic notch was outlined. These contours were uploaded into SHAPE v1.3 and subjected to elliptical Fourier analysis. Principle components generated eigenvalues that identified the variables contributing to the shape of the notch, and 97% of the observed variation could be described using four principle components. Notch shape varies systematically with each of the aforementioned variables, with sex having the greatest contribution. As expected, females display wider symmetrical notches, while those of males are narrow and asymmetrical. However, the final model was only able to explain 14.3% of the total variation in the shape of the notch, indicating that the growth and development of this feature is quite complex.
Genome Wide Association Study of Dental Arch Form and Occlusal Relationships in the Mixed Dentition Stage
Authors: Miller Steven , Shaffer John , Marazita Mary , Levy Steven , Warren John , Hartsfield James , Moreno Lina
Published: June 2021

Dental arch form has been a major subject of investigation in the dental sciences, however, little is known about the genetic etiology of arch form and arch occlusal relationships. This study seeks to examine the potential genetic underpinnings of dental arch form and malocclusion. A sample of n=272 dental casts of the mixed dentition stage (around age 9) were scanned and landmarked to capture data pertaining to dental arches shape and position and then subjected to principal components analysis (PCA) to identify the most important aspects of phenotypic variation. These PC scores were then correlated with whole‐genome association data for a subset of n=210 individuals to identify potential relationships between dental arch form and the genome. Phenotypic variation captured through PCA identified antero‐posterior (AP) discrepancies between the maxillary and mandibular arch (PC1 – 24.2% variation) as well as arch width variation (PC2 – 15.8%) and AP discrepancies with variation in the curve of Spee (PC3 – 14.9%). Suggestive relationships between PRKD1 and PC1 (p=7.11E‐06), RBFOX1 and PC2 (p=8.06E‐06) as well as DACH1 and PC3 (3.27E‐06) were found. This study is among the first to identify variation in dental arch shape and position that may be related to variants in these genes.

Support: Carver Charitable Trust #12‐4058 and NIH 2 UL1 TR000442, 1T90DE023520, R01‐DE09551 and U01‐DE018903
Bilateral Asymmetry of the Foramen Ovale: Neurosurgical Implications
Authors: Cyrus Leah , Laslo Jillian , Zdilla Matthew
Published: June 2021

The foramen ovale (FO) is located in the middle cranial fossa and found anteromedial to the foramen spinosum. The FO transmits numerous anatomical structures such as the mandibular branch of the trigeminal nerve (V3), the accessory middle meningeal artery, and the lesser petrosal nerve. The FO is clinically important, particularly with regard to surgical procedures that cannulate the foramen. Such procedures include transovale treatment of trigeminal neuralgia and biopsy of parasellar lesions. The morphology of the right and left FO of 84 adult skulls were compared. The maximum diameter, area, and perimeter of the right FO were significantly larger than those of the left FO. Knowledge of the asymmetry between the right and left sided FO is important in preventing improper cannulation and subsequent intracranial hemorrhage. Research was supported by funding from the WV Research Challenge Fund [HEPC.dsr.14.13].
Mechanical effects of fine‐wire climbing on the hindlimb skeleton of mice
Authors: Joll J. , Vickery B , Rupert J. , Biro K. , Wallace J. , Byron C , Organ J.
Published: June 2021

High‐impact exercise (running/jumping) can stimulate multiple anabolic responses (increased trabecular bone volume, BV/TV) in the skeleton, but is also linked to an increased incidence of skeletal fracture. Thus, it is not an appropriate treatment for patients with elevated fracture risks. However, multi‐directional off‐axis mechanical loading can also elicit anabolic responses, even when magnitudes are relatively low. This represents a potential alternative to high‐impact exercise for improving skeletal mechanical properties. To test this hypothesis, we raised twelve weanling female C57BL/6 mice to 4 months of age in custom enclosures that prevent (control) or require (experimental) manual and pedal grasping while balancing and climbing above narrow wire substrates. At sacrifice, we measured whole mouse bone density (DEXA) and performed architectural (μCT) and mechanical (4‐pt bending) analyses of the femur and tibia. Body mass was similar between groups, although exercised mice were leaner (‐35% fat mass). Bone mineral density was also similar, while bone mineral content was increased (+7%) in the exercised mice. Femoral midshaft polar moment of inertia was similar between groups, but exercised mice had lower BV/TV (‐46%) of the distal femur and greater trabecular spacing (+21%). Exercised femora showed more total displacement (+58%) and post yield displacement (+115%) in bending than controls, and increased material toughness (+40%). Patterns were similar for the tibia. Mechanical data are consistent with high‐impact exercise studies, but architectural data are not. Together they suggest that our exercise model may improve bone mechanical properties by redistributing mineral within the skeleton, and not by increasing net bone formation.
APOPTOSIS IN BONE OF DIABETIC RATS TREATED WITH LOW INTENSITY LASER
Authors: Silva Jairo , Fazan Valéria , Tirapelli Daniela , Thomazini José , Tirapelli Luis
Published: June 2021

Apoptosis is mechanism of programmed cell death mechanism. Diabetes Mellitus is a chronic syndrome, and decreases blood flow and consequently the time of consolidation. The therapy with the low intensity has show efficiency in cellular regeneration and can accelerate the bone healing process of immune response/inflammatory, which play an important role for the regeneration of bone tissue (Baxter, 1994). This study aimed to evaluate the efficacy of diode laser 904nm in regeneration bone in diabetic rats. Sixty male Wistar rats were divided into 6 groups and submitted to bone defect on the left tibia. The Group 1 was not induced diabetes and not treated with laser; Group 2 was not induced diabetes and treated with laser dose 50w to 7 applications; Group 3 was not induced diabetes and treated with laser 100w with the same protocol; Group 4 induced diabetics with streptozotocin and the thent day after undergoing the surgical procedure but not treated with laser; Group 5 induced diabetes and treated with laser dose 50w/cm; Group 6 induced diabetes and treated with laser 100w with the same protocol. We used immunohistochemistry to analyze expression of CASPASE‐3 and and Computer Tomography for bone parameters. The results shows decrease of CASPASE‐3 in groups treated with laser and X‐rays and bone densitometry suggest that the 904 nm laser diode is to improve bone repair.
Sexual Dimorphism in The Birth Canal and Non‐Obstetric Pelvis among Anthropoids: Effects of Cephalopelvic Proportions
Authors: Moffett Elizabeth , Ward Carol
Published: June 2021

As important as birth is hypothesized to be in shaping the primate pelvis, we actually know very little about how birth affects pelvic form across anthropoids. This study tests whether large cephalopelvic proportions are related to dimorphism in the obstetric and non‐obstetric pelvis. Landmark data for 118 anthropoid specimens were analyzed [Large cephalopelvic proportions: Ateles geoffroyi (n=4), Homo sapiens (n=40), Hylobates lar (n=22), Macaca fasicularis (n=10), Nasalis larvatus (n=13), Papio cynocephalus (n=10), Saimiri sciureus (n=38); Small cephalopelvic proportions: Alouatta palliata (n=5), Gorilla gorilla (n=21), Pan troglodytes (n=20), Pongo pygmaeus (n=12)]. Birth canal dimensions were significantly larger in females compared to males (p<0.05) within species having large cephalopelvic proportions, but not in species with small proportions. Birth canal shape, quantified using principal components analysis, was also dimorphic in species with large cephalopelvic proportions (all p‐values <0.05), but not for species with small proportions. Patterns of dimorphism, quantified using Euclidean Distance Matrix Analysis, were shared among phylogenetic groups (New World monkeys, Old World monkeys, apes). Finally, dimorphism in the non‐obstetric pelvis did not correspond predictably to the presence of birth canal dimorphism or to large cephalopelvic proportions.
Treating Bone Quality in Chronic Kidney Disease
Authors: Newman Christopher , Hammond Max , Wallace Joseph , Tian Nannan , Chen Neal , Moe Sharon , Allen Matthew
Published: June 2021

While patients with chronic kidney disease (CKD) experience significant bone loss, little is known about the role of bone quality (properties independent of size and shape). Calcitriol is used to prevent bone loss by inhibiting parathyroid hormone (the main cause of bone breakdown in CKD), yet its impact on bone quality is unknown. Raloxifene improves bone quality, but its effects in CKD are unknown. This study examined the hypothesis that bone quality is deteriorated in CKD and that raloxifene but not calcitriol would improve bone quality. Using a rat model of CKD, mechanical properties of the tibia were assessed by nanoindentation, and composition was assessed by Raman spectroscopy. Animals with CKD were not different from normal controls with regard to elastic modulus, hardness, mineral‐to‐matrix ratio, or mineral crystallinity. Calcitriol treatment resulted in a higher mineral‐to‐matrix ratio (+20%), but it did not improve mechanics. Raloxifene, on the other hand, did not impact bone composition but led to substantially higher tissue hardness (+82%), consistent with previous data showing a mechanical effect independent of changes in tissue composition or bone mass. While animals with CKD may not have significant defects in bone quality at this stage of disease, these results indicate that improving the quality of the remaining tissue may be an effective means to compensate for bone loss in CKD.

Nanoindentation
Raman Spectroscopy

Hardness (MPa)
Elastic Modulus (GPa)
Mineral‐to‐Matrix Ratio
Mineral Crystallinity

Normal (n=8)
196.97 +/‐ 23.53
11.13 +/‐ 2.57
2.69 +/‐ 0.44
0.053 +/‐ 0.0009

Chronic Kidney Disease ‐ No Treatment (n=9)
212.15 +/‐ 52.79
10.63 +/‐ 1.66
2.30 +/‐ 0.48
0.054 +/‐ 0.0008

Chronic Kidney Disease ‐ Calcitriol (n=11)
209.43 +/‐ 47.29
9.99 +/‐ 2.51
2.76 +/‐ 0.36 (*)
0.053 +/‐ 0.0004

Chronic Kidney Disease ‐ Raloxifene (n=10)
386.84 +/‐ 282.29 (*)
12.64 +/‐ 3.31
2.29 +/‐ 0.45
0.053 +/‐ 0.0005
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.
A Histological Examination of the Lumbar Sympathetic Trunk
Authors: Mintz Noah , Beveridge Tyler , Sandig Martin , Johnson Marjorie , Allman Brian
Published: June 2021

Current gross anatomical and histological descriptions of the lumbar sympathetic trunk suggest that its neuronal cell bodies are located in discrete ganglia that are linked to each other by nerve fibres. Despite well‐documented variations in the macroscopic morphology of the lumbar sympathetic trunk (e.g., number and placement of ganglia), a detailed histological examination of the arrangement of the neuronal cell bodies within the trunk is lacking. Thus, the present study aims to examine the microscopic organization of the lumbar sympathetic trunk using standard histological techniques. In our preliminary investigation, a 10 cm length of the lumbar sympathetic trunk (including ganglia and interganglionic fibres) was serially‐sectioned (5 μm thick) in the longitudinal plane and then stained with Haematoxylin & Eosin to visualize the location of neuronal cell bodies throughout the 10 cm length. This examination revealed that the neuronal cell bodies are not confined to specific “ganglionic” swellings; a finding which is inconsistent with the current textbook descriptions of the lumbar sympathetic trunk. The presence of neuronal cell bodies throughout the “interganglionic” regions of the lumbar sympathetic trunk may have implications for future investigations of sympathetic activity in this region, specifically related to understanding functional deficits associated with this nerve tissue.
Deltoid Muscle: Patterns of Innervation from Axillary Nerve
Authors: Lima Larissa , Itezerote Ana , Hojaij Flávio , Andrade Mauro , Jacomo Alfredo , Akamatsu Flávia
Published: June 2021

Introduction

The 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 occurrence

Study objective

Determine 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 Methods

Two 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 branches

Taking 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 Results

Preliminary 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.

Conclusion

Our data can help avoiding iatrogenic injuries during procedures and, they also help understanding the myofascial pain and its treatment.
Innervation to Mandibular Gingiva
Authors: Dorius Geoffrey , Fandino Luis , Sanders Luke , Clayton Craig , Edmunds Kinnison , Labinpuno Vince , Nixon Anthony , Wisco Jonathan , Morton David
Published: June 2021

General sensory innervation of the mandibular gingiva by branches of the mandibular nerve (CN V‐3) is well documented. However, general sensory branches from the lingual nerve that traverse the submandibular gland have not been well documented. Therefore, using a combination of macroscopic dissection, MicroScribe 3DX, and Sihler's nerve stain, data was gathered as to which branches of the lingual nerve traverse the submandibular gland on route to innervating the mandibular gingiva. In 30 dissected specimens, the lingual nerve enters the oral cavity near the postero‐lateral surface of the tongue and sends posterior branches into the lingual mandibular gingiva, posterior and medial to the third molars. Other small branches innervate the sublingual gland and the tongue. Anterior nerve branches emerging from the sublingual gland innervate the lingual mandibular gingiva. Altogether, there are an average of 2.62 branches to the mandibular gingiva. The data demonstrated lingual nerve branches that are not presently documented in atlases, and journals. A continuation to collect specimens will allow for a greater amount of information regarding the number of branches from the parent lingual nerve innervating the mandibular gingiva, along with the consistency of how many nervous branches are found. This information could be helpful to dentists in performing local anesthetic blocks.
Relation Between the Conduction Velocity of Nerve Fibers, Micro‐anatomical Parameters and the Divine Proportion (Φ=1.618) ‐ A new Paradigm
Authors: Bonatto‐Costa João , Thomaz Leonardo , Goulart Guilherme , Xavier Léder , Rocha Andrea , De Campos Deivis
Published: June 2021

The Divine Proportion (DP) (Φ=1.618) is found in several natural phenomena and in different human anatomical proportions, such as cephalometric measurements, anatomical organization of the hands and functional and esthetic organization of dentition. Recent studies have described that structures or systems organized based on DP are more stable, aesthetically more pleasurable and functionally more eficiente as they follow mathematically the biological laws. The aim of this study was analyze the DP and other micro‐anatomical parameters in a pair of nerves with similar function, but with different velocities of conduction of nerve impulse ‐ the recurrent laryngeal nerves (RLN) left and right ‐ in which the right RLN presents a larger velocity of nerve impulse conduction. Seven RLN of cadavers obtained from necropsies were analyzed with mathematical, stereological and images tools using the Software Image Pro‐Plus Software 6.0. The micro‐anatomical comparison between the right and left RLN showed that the following parameters of the right RLN are significantly larger: axon diameter (11.2%, p=0.001) and myelinated fiber diameter (20.6%, p=0.0001). The left RLN had a G‐ratio significantly larger than the right RLN (10.3%, p=0.0093). There is a significant difference in DP between the right and left RLN (p=0.0047), in which the right RLN has a perfect value of DP (Φ=1.6), while the left RLN has a value of Φ=1.4. Possibly, the conduction velocity of nerve impulse may not be only affected by the presence of myelin sheath and by axon diameter, but also by “perfect” morphology of nerve fiber.
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 35,55

Support: 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
Preventing Alzheimer's Pathology in a Preclinical Mouse Model
Authors: Shaw Phyllis
Published: June 2021

More than five million people live with Alzheimer's disease (AD) in the U.S. today and the number is rapidly rising. This disease robs its victims of dignity and places untold burden on families and on the economy. Cystatin C (CysC) is highly expressed in normal brain, but drops at the onset of AD and remains low during AD progression. Forced expression of CysC lowers Aβ brain deposition in AD mouse models. Since CysC protein can both inhibit amyloid precursor protein (APP) degradation by cathepsin B and prevent Aβ fibril formation in vitro, we synthesized the Aβ‐binding portion (CysC‐AβBP), labeled it with FITC and showed that it binds specifically to Aβ42 oligomers, that it can be efficiently delivered to the brain intranasally and that, when intranasally delivered into an AD mouse model, it binds to Aβ in the hippocampus and the neocortex. In our in vivo studies, 2.7 and 4 month old CRND8 AD mouse littermates were treated intranasally with 20 micrograms of CysC‐AβBP or scrambled CysC‐AβBP peptide, twice weekly for 3 weeks. Sagittal sections of brains from untreated and treated AD mice were stained with Aβ‐specific antibody 6E10 (Covance) and Texas Red‐conjugated secondary antibody. The brains of AD mice with no treatment or scrambled peptide treatment had a large accumulation of amyloid beta aggregates and plaque in the neocortex and hippocampus. Importantly, a 3‐week treatment of AD mice with CysC‐AβBP reduces Aβ aggregates and plaque up to 80% in the hippocampus and neocortex.
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 purpose

Methylene 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.

Methods

Rats 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.

Results

The 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).

Conclusions

MB induced neuroprotection,at least in part,by enhancing autophagy and reducing apoptosis in perfusion‐diffusion mismatch tissue following ischemic stroke.
The “Analytical” Brain by Auzoux, An Anatomical Model in Papier‐mâché, Circa 1870: Description And Photographical Tri‐Dimensional Reconstruction
Authors: LE FLOCH‐PRIGENT PATRICE , GILLOT JEAN‐BERNARD
Published: June 2021

The « analytical » brain was made by the Auzoux’ factory in papier mâché from 1867. It was in accordance with the anatomical and neurological knowledge of this times. It represented the very enlarged structure of the brain and of the brain stem (40cm high) and was specially devoted to the white matter. The grey matter was only a support and an anatomical relationship on the right aspect. The dismantable character of the sample was limited to a separation in a few pieces which detailed the internal capsule and the grey central nuclei of the left hemisphere. This model was rare and spectacular. By one hand it was the prefiguration 140 years in advance of the tractography which recently became possible on the living person by the new imaging methods. The reduced (real size) model is more often encountered.
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
Repairing Injured Optic Nerve of Rat with Several Therapies
Authors: Xu Jiajun , Liu Fang , Xie Jiaxin , Wang Ruobin , He Junshan , Qi Aiqin , Cao Wenluo , Sun Jingchuan , Xu Tao , Yuan Jianming
Published: June 2021

Optic nerve of mammalian, as brain and spinal cord, is difficult to regenerate successfully after injury. It is one of research focuses and difficult projects how to promote regeneration of injured optic nerve in the neuroscience and ophthalmology. We have explored to repair the injured optic nerve of rat with several therapeutic strategies in the recent years. (1)Physical therapy: In a rat model of optic nerve crush, hyperbaric oxygen preconditioning or hydrogen‐rich saline administered intraperitoneally could decrease apoptosis of retinal ganglion cells, promote their survival, and improve visual functional restoration as pupillary light reflex and visual evoked potential. (2)Cell therapy: Rat amniotic epithelial cells (AECs) with some characteristics of stem cells, AECs transfected by basic fibroblast growth factor gene, or AECs seeded in the collagen sponge was implanted into the injured region of rat optic nerve transected. These three therapies could increase the survival of retinal ganglion cells after injury of rat optic nerve. The cells could guide the regenerating axon to extend into the injured region, and promote a few of axons to cross the injured region into 1‐2 mm in the distal optic nerve. (3)Gene therapy: After neuritin lentivirus infection of rat retina, the number of retinal ganglion cells and optic nerve axons were increased significantly after optic nerve crush, this protective effects was further confirmed by the functional tests as pupillary light reflex and visual evoked potential. After RGC5 damage in vitro, neuritin overexpression promoted neurite outgrowth which might be through the upregulation of phosphorylation of Akt and Erk, and reduced cell apoptosis which might be through the inhibition of caspase 3 activation.
Ultrastructural Changes in the Optic Nerve After Injury and Neurotrophic Factor Treatment
Authors: Vega Giam , Blagburn Jonathan , Blanco Rosa
Published: June 2021

We have shown that ciliary neurotrophic factor (CNTF) and fibroblast growth factor (FGF‐2) have strong facilitatory effects on axon regeneration in the adult frog optic nerve after injury. In these nerves, bundles of regenerating axons were associated with astrocytes and macrophage‐like cells. The objective of the present study is to characterize the identity of these cells, and to determine the changes that occur after CNTF or FGF‐2 application. We performed optic nerve crush and applied into the nerve either saline solution or neurotrophic factors. We examined the optic nerves at different timepoints: 48h, one week, and two weeks after axotomy. Electron microscopy was used to characterize the ultrastructure and localization of the macrophages/microglia after injury and neurotrophic treatment. Antibodies against specific macrophage subtypes were used to label these cells in confocal images of different regions of the nerve. We found a large number of macrophage‐like cells at the lesion site, and distally in close proximity to regenerating axons in CNTF and FGF‐2 treated nerves. Significantly fewer cells were present proximal to the lesion. Many of these macrophage‐like cells at, and distal to, the lesion were ED‐1‐positive in both CNTF and FGF‐2 treated nerves. Both M1 (Arginase‐positive) and M2 (CD‐86‐positive) subtypes were identified. We are currently quantifying the distribution of these cell subtypes at different timepoints and nerve regions after injury. In conclusion, the application of CNTF and FGF‐2 affects the number and the distribution of macrophage subtypes after optic nerve injury and during optic nerve regeneration.
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).
Identification of a Resilient Population of Axotomized Motoneurons within the Mouse Facial Motor Nucleus
Authors: Meadows Rena , Muldoon Jessica , Haulcomb Melissa , Beahrs Taylor , Schartz Nicole , Olmstead Deborah , Batka Richard , Sanders Virginia , Jones Kathryn
Published: June 2021

Previous facial motoneuron (MN) survival studies have identified a facial MN population that is resilient to axotomy. Examination of the literature suggests two distinct populations of MN: one population vulnerable to axotomy and another population potentially resilient to axotomy. To identify potential differences in MN survival levels after target disconnection, we performed a facial nerve transection at the stylomastoid foramen in a number of different mouse strains, as well as models of immunodeficiency and MN disease. All mice were monitored for facial MN survival for up to 26 weeks post‐axotomy (wpa). We observed an 85% survival of facial MN at 4 wpa that was reduced to about 50% at 10 wpa in WT mice. In contrast, facial MN survival significantly dropped at 4 wpa to approximately 62% and 50% in immunodeficient and MN disease mice, respectively. Similar to WT mice, facial MN survival levels remained at about 50% for up to 18 wpa in both the immunodeficient and MN disease mice. Collectively, these data suggest that, despite target disconnection of similar MN populations by either axotomy or disease, there are inherent differences that may be exploited for therapeutic applications in the future.
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.
Synthesis and Characterization of Deoxyribonucleic Acid Aptamers for β‐D‐Glucan Carbohydrates
Authors: Webster Nicholas , Rohlman Christopher
Published: June 2021

This work explores the use of short randomized single‐stranded DNA aptamers which have the ability to be selected to bind to molecular structures including DNA, RNA, protein, and carbohydrates. This work will focus on the selection of aptamers that show selectivity for the β‐D‐Glucan carbohydrate, a polysaccharide that has many roles in immunity, cholesterol metabolism and infection. This work will act as proof of concept for targeting carbohydrates for future studies.
Measurement and Detection of Mitochondrial DNA Deletions in Irradiated Human Cell Lines
Authors: Gautam Shikha , Gulfo Michelle , O'Donnell Robert , Pogozelski Wendy
Published: June 2021

The effect of ionizing radiation such as that of X‐rays on mitochondrial DNA is not well‐understood. We have been using PCR‐based methods to quantify and detect changes in human mtDNA in cell culture. We previously developed a real‐time PCR assay for measuring numbers of mitochondrial DNA copies using either plasmid standards into which mitochondrial DNA had been cloned or isolated human mitochondrial DNA as quantitative standards for mitochondrial DNA genomes. Now we investigate a different real‐time PCR quantification method using single‐copy nuclear genes as quantitative standards for cell number. We also show how x rays affect the amount of total mitochondrial DNA and mitochondrial DNA bearing the 4977‐bp common deletion. In addition, we show how long‐extension PCR (LX‐PCR) can reveal the presence of new deletions in human lymphoblasts treated with X‐ray doses up to 5 Gy.
Examining KIT, EDNRB, MITF, PMEL17, MLANA Genes to Determine the Genetic Basis of Equine Deafness
Authors: Achimovich Alecia , Innamorati Katherine , Lipsett Koren
Published: June 2021

There appears to be a correlation between congenital deafness and lack of pigmentation in a small pedigree of Spanish Mustangs. At this time, no hereditary explanation has been presented for this pattern as it appears in horses. Based on the previously determined correlation of melanocyte function and/or migration to deafness in humans, we have conducted a candidate gene study to attempt to link the white coat pigmentation, blue eyes and cochleosaccular deafness within this pedigree. Candidate genes were chosen on the basis of correlation to similar disease states in humans, dogs and cats, including KIT, EDNRB, MITF, and MLANA. KIT, a receptor tyrosine kinase in the pigment production pathway, has been implicated in multiple mammalian deafness studies to date. A mutation (KI16 + 1037A) in the 5′ splice junction of exon 17 is known to cause sabino coloration, but was determined to not be linked to deafness. Alternatively, a heterozygous glycine to serine non‐synonymous mutation was identified within exon 6 of MITF, a transcription factor necessary for proper melanocyte differentiation, but doesn't appear to correlate to deafness. In addition, no mutations within exons were found in neither EDNRB (endothelin receptor type B) nor MLANA, also known as melan A or MART1 (melanoma antigen recognized by t‐cells). Further analysis of additional candidate genes, including PMEL17 and SOX10, and the inclusion of intronic sequence analysis for all candidate genes, will be necessary to determine the cause of cochleosaccular deafness, through mutations in these genes or others.
Mechanisms of cytotoxicity for bifunctional epoxide alkylating agents
Authors: Redstone Samuel , Ahern Darcy , Scott Paul , Le Phuong , Millard Julie
Published: June 2021

The discovery that bifunctional alkylating agents have antitumor activity led to the development of cancer chemotherapy over 50 years ago, yet these compounds also lead to an increased risk of secondary cancers. Bifunctional alkylating agents form a variety of DNA adducts, including monoadducts, intrastrand cross‐links, and interstrand cross‐links. Interstrand cross‐links are believed to be the most lethal, impeding both replication and expression of the genetic material. We are characterizing the mechanisms by which diepoxybutane (DEB) and epichlorohydrin (ECH) exert their cytotoxic effects in cultured cells. Our first goal is to determine the relationship between interstrand cross‐linking and cytotoxicity in human acute myeloid leukemia (HL60) cells. HL60 cells are treated with varying concentrations of the compound of interest and assayed to determine the fraction of cells that remain viable, allowing determination of LD50 values. Cross‐linking ability is assessed using an ethidium bromide assay to determine the amount of duplex DNA following alkaline denaturation. Only cross‐linked DNA reanneals rapidly and interacts with the dye, which is highly fluorescent when bound to duplex DNA. Preliminary results suggest a correlation between cross‐linking and LD50 values for these compounds, with DEB a more efficient cross‐linker than ECH.Our second goal is to identify the structures of ECH‐cross‐linked lesions via ESI‐TOF‐MS, which may provide information about the mechanism of this agent. This research was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103423.
Determination of the HTLV‐1 pro‐pol Frameshift Site Secondary Structure
Authors: Durnford Kathryn , Chapman Erich , Atene Antonia , Broad Amanda , McKenzie Jason , Yeager Dan , Kieft Jeffrey , Mouzakis Kathryn
Published: June 2021

Expression of human t‐cell leukemia virus type I (HTLV‐I) enzymes requires two ‐1 programmed ribosomal frameshifts. These events occur between the gag‐pro and pro‐pol open reading frames. Each frameshift site includes a heptanucleotide slippery sequence followed by a downstream structure, which act in cis to produce specific frameshift efficiencies. While ‐1 PRF and the slippery sequences of these frameshift sites have been established in HTLV‐I, their frameshift efficiencies and the structures within these sites have not been determined. In pro‐pol frameshift site, an RNA pseudoknot is predicted to fold downstream of the UUUAAAC slippery sequence. However, no structural data exist for this RNA. Here, we report a preliminary structure of the HTLV‐1 pro‐pol frameshift site RNA. Nucleotide reactivity data acquired from selective 2′‐hydroxyl acylation experiments analyzed by primer extension and thermodynamics‐based secondary structure predictions are consistent with a pseudoknot secondary structure. Additionally, non‐denaturing PAGE analysis of the wild‐type structure and two mutant RNAs confirmed that the fold of the wild‐type RNA is significantly different from RNAs of identical length that cannot form a pseudoknot structure. These results establish the existence of a pseudoknot structure in the HTLV‐1 pro‐pol frameshift site.