bluuebunny/crossref_metadata_2025 huggingface dataset:
Targeted Proteomics: Endothelial Cell Membrane Response to TNF‐α
Authors: Prisco Anthony , Hoffmann Brian R , Kaczorowski Catherine C , Greene Andrew S
Published: June 2021

Endothelial cells play a key role in regulating vascular function. One key role of the endothelium is to control the inflammatory response, limiting it to acute periods of defense. Produced by macrophages, one of the first cytokines produced in the inflammatory response is tumor necrosis factor alpha (TNFα). TNFα signaling to endothelial cells is essential for inducing the expression of proteins that recruit additional cells of the inflammatory response. TNFα signaling is complex, having two receptors that each signal through multiple pathways. In this study, we determine the effect of low dose TNFα on differential membrane protein expression in endothelial cells, identifying cell surface proteins that are upregulated, downregulated or uniquely expressed.

We developed a novel proteomics technique, cell membrane associated enrichment (CMAE) for analyzing proteins contained on the outer cell surface and those closely associated with the cell surface. CMAE, targeting plasma membrane proteins with a surface amine group, combined with a more well‐established surface N‐glycoproteome enrichment (CSC) allowed increased coverage of the cell surface proteome. After CSC and CMAE results were combined allowing us to specifically target differentially regulated plasma membrane proteins in that were then validated by flow cytometry analysis of endothelial cells.
Workshop Format Increases Scientific Knowledge, Skills, and Interest when Implemented in an Introductory Biology Course that Attracts and Retains Underrepresented Minorities
Authors: Johnson Kathryn M.S. , Lownik Joseph C.
Published: June 2021

Human Biology, an introductory biology course for majors and non‐majors, taught in a workshop format, integrates hands‐on laboratory activities, group work, discussion, and lecture to teach core biological principles and skills in critical thinking and communication. Human Biology attracts students that identify themselves as underrepresented minorities (URM) at a proportion greater than that observed college‐wide (24 ± 4% vs. 13 ± 2%), and overall student retention in the course is ~94%. To determine student gains in core biological concepts and competency in critical thinking and communication skills, students that were enrolled in four sections of Human Biology self‐assessed their knowledge and skills. Students were surveyed prior to (pretest, n = 63) and after (posttest, n = 75) the semester‐long course. There was a significant increase (P ≤ 0.05) in self‐reported conceptual understanding of biological knowledge and critical thinking and communication skills. When asked how the format of the course assisted in their learning, 80% of posttest respondents indicated “a lot” or “a great deal.” Posttest respondents also indicated that work in this course resulted in “great” gains in interest in taking additional science courses and majoring in a science‐related field. Therefore, Human Biology, a course that attracts and retains URM, increases student‐perceived knowledge, skills, and interest in science.
Evaluating the effectiveness of the “flipped” or “blended” Physiology classroom for JAMP students
Authors: Lee Osborne David , Janssen Herbert F
Published: June 2021

“Flipping” a classroom is accomplished when students are provided with information before class and face time is used to develop conceptual learning. The pre‐class assignments often use recorded lectures or “voice over” PowerPoint presentations to emphasize cognitive information normally found in the lower strata of Bloom's taxonomy. During class, students are expected to use this basic information to develop a more in‐depth understanding of the material through deliberate practice. The current study developed an assessment tool that could be used to evaluate the success of the flipped classroom. Three topic areas were chosen as part of a summer preparation course for Joint Admission Medical Program (JAMP) students. Material was presented related to the Physiology of body fluids, respiration and bone. For each topic, lecture, discussion and laboratory sessions were designed. The order of the three types of classroom activities was varied by order of presentation. Students were given the first test before any class activity was performed and averaged 5.61/12 questions correct. Exposure of students to lecture or discussion yielded a 49% increase in question success. However, exposure to laboratory based exercise yielded only a 6 % increase in question success. Results suggest that such an assessment procedure promoted learning and was well received by the students.
Team‐based learning format in the medical physiology teaching laboratory
Authors: Schmidt John William
Published: June 2021

Case‐based problem solving that is structured according to the Team‐Based Learning (TBL) format (www.teambasedlearning.org) is being used in a first year medical school curriculum that combines anatomy, physiology and biochemistry in a single Human Biology course.

Objective

how effective is the TBL format when applied to multi‐disciplinary lab exercises? One lab exercise allows students to explore the role of alkaline phosphatase in the control of intestinal lumen pH. This exercise combines enzymology techniques with learning of physiology concepts related to homeostasis, receptor function, signal transduction and regulation of epithelial ion transport. This lab exercise has previously been taught to four student cohorts in a conventional group work format. In the TBL format, student understanding of the assigned reading is assessed at the start of the lab session by a multiple‐choice Individual Readiness Assessment Test (I‐RAT) using an electronic student response system. Teams then discuss their quiz answers while completing a Group RAT before the collection of data using a colorimetric enzyme assay. Most of the lab session group work is data collection followed by answering questions about the data and additional reading in two published journal articles. Effectiveness of the TBL format will be assessed by comparison of lab report scores and performance on multiple‐choice exam questions.
Translational anatomy research at medical and undergraduate universities
Authors: Wisco Jonathan J. , Stark M. Elena , Lambert H. Wayne
Published: June 2021

As a foundational science, anatomy is well positioned as a discipline to address the NIH Roadmap for Medical Research (2004) calling for developing new pathways to discovery, establishing multi‐disciplinary research teams of the future, and re‐engineering clinical research enterprise. We highlight our successful interdisciplinary collaborations formed with clinicians and basic scientists that led to, or augmented, “new” anatomical sciences research in the fields of neuroimaging, neurology, radiology, cardiology, anesthesiology, surgery, orthopedics, acute care, head and neck surgery, physical therapy, and pain management. Many of our collaborations either started as teaching innovations then evolved into basic science research, or vice‐versa. Indeed, the gross anatomy and microscopic anatomy laboratories are essential venues for establishing collaborations. Finally, we discuss the value of the anatomical sciences to a medical school and undergraduate university's effort to bolster translational research and innovative education.
DEMONSTRATION OF LEFT GASTRIC ARTERY VARIATIONS BY MULTISLICE CT ANGIOGRAPHY
Authors: SHARMA PRADEEP KUMAR , SEHGAL GARIMA , SRIVASTAVA AJAI KUMAR , SINGH RAGINI , PARIHAR ANIT , AGA PALLAVI
Published: June 2021

To identify and evaluate the normal and variant anatomy of left gastric artery (LGA) by computed tomography a cross sectional study was carried out in the Departments of Anatomy and Radiodiagnosis, K G Medical University, U.P, Lucknow, India. CT‐angiograms of 50 subjects were analysed. The left gastric artery was found to be the smallest branch of the celiac trunk and displayed anomalous origin and numerical variations. Replaced origin of LGA was seen in 9 subjects. In 6 subjects LGA shared its origin with inferior phrenic artery and both were arising from a common trunk. Single LGA was present in majority of subjects (96%) whereas the remaining 4% had two LGAs. Accessory LGAs were found to be the branches of left hepatic artery. They traversed the fissure of ligamentum venosum to reach the stomach. Knowledge of such variations can influence surgical procedures and the planning of image‐guided interventions.
A quantitative, biomechanical analysis of radioulnar deviation between fresh frozen and soft embalmed human forearms
Authors: Casier Craig Bradley , Easteal Ron , Sellens Rick , Dickinson Andrew , Clark Jessica
Published: June 2021

Fresh frozen human cadaveric specimens are considered the gold standard for biomechanical research; however, the testing period of these specimens is limited because they decay rapidly, can be difficult to obtain and can carry infectious diseases. Therefore, various chemical fixation methods to deal with these issues have been developed. Traditional formalin fixation has been shown to significantly affect the biomechanical properties of the tissues; however, a relatively new method called soft embalming could provide an alternative to fresh frozen specimens in the field of biomechanical research. The purpose of this pilot study was to determine the effects soft embalming has on gross anatomical movements. The parameters for testing were radioulnar deviation range of motion with in plane and out of plane angulation measured using an incremented tendon loading protocol. Five fresh frozen human cadaveric forearms were obtained and injected with tantalum tracer beads and tested pre and post embalming using 3D X‐rays. The results showed that the soft embalmed specimens retained their flexibility and was slightly greater than the fresh frozen state. Although, at lighter loading trials, the range of motion in soft embalmed specimens was larger than the fresh frozen state. The results here may provide an argument for the inclusion of soft embalmed specimens in biomechanical testing.
GREAT CELOMIC CAVITY : AN ANATOMICAL MODEL IN PAPIER MACHE, 20th CENTURY
Authors: LE FLOCH‐PRIGENT Patrice
Published: June 2021

A real sized, anatomical model in papier‐mâché of the great celomic cavity in an adult belonged to the GILLOT‐BRIEUX scientific bookstore (48 rue Jacob, 75006 PARIS). It was probably designed and built in the first half of the 20th century in Europe. Neither trade mark, nor date were found. It laid on a huge basis in wood (45×90 cm) painted in black on the visible aspects. A cloth surrounded the inferior part of the trunk. The anterior aspect of a complete dissection of the thorax, abdomen and pelvis was represented. The right lung was frontally cross‐sectionned and the main mediastinal vessels were represented. At the abdominal level, the emphasis was on the retro‐peritoneal region with the urinary system (kidneys, ureters) and the adrenal glands. The muscles of the (thoracic and abdominal) walls were detailed on frontal cross‐sections. Quite certainly, the purpose of this model was the teaching in the universities (medical schools).
Chicken or the egg?: The incidence of lower limb pathology and the association with individuals with increased body mass
Authors: Kelso Rebecca Scopa
Published: June 2021

Clinically, it is important to understand the long‐term mechanical effects of increased loadbearing on joints of the lower limb throughout ontogeny. This study looks at relationships among increased mass, femoral and proximal tibia dimensions, and the differences between individuals with and without lower limb skeletal pathology. A clinical control sample of juveniles categorized as having normal mass, both with and without skeletal pathology, was compared with juveniles considered clinically obese, also with and without skeletal pathology. Radiographic measurements of the femoral head, distal femur, and proximal tibia were taken. Measurements obtained from the images were corrected for parallax. Results indicate a high correlation between obese individuals and increased dimensions from femoral head and the medial knee, as well as greater frequencies of pathology. Furthermore, there is some indication that joint sizes may be adaptive and responding with skeletal pathology to increased weight gain in older children (10–17 yrs). This may indicate overcompensating in joint size growth among obese juveniles, most likely in response to high mechanical loads. The limitations of the data leave the timing of the onset of obesity unknown, however, and therefore the ontogeny of joint hypertrophy cannot yet be established.

Grant Funding Source: N/A
Effectiveness of the 904nm laser diode for regeneration of bone defects.
Authors: Silva Jairo Pinheiro , Fazan Valéria Paula Sassoli , Pardini Luís Carlos , Thomazini José Antônio
Published: June 2021

Acceleration of bone formation for repair of large bone defects is one of the great challenges in health science. Several studies are investigating mechanisms for bone repair on a quick and efficient manner and it is well known that the modulation of the immune/inflammatory responses play an important role for the bone tissue regeneration. This study was designed to evaluate the efficacy of 904nm Laser Diode in the bone tissue regeneration to repair a critical bone defect. Thirty male Wistar rats were separated into 3 groups of 10 animals each. Control rats were submitted to a circular bone defect with a 2 mm drill on the left tibia. A second group, submitted to the same bone defect as the controls, was irradiated daily with a 904nm laser diode, 60w 4J/cm2, from the first post‐operatory day up to 14 days. A third group was irradiated with 904nm laser diode, 120w 4J/cm2 using the same protocol described for the second group. After 14 days, the bone defect was assessed by means of X‐ray and bone densitometry. Animals were then anesthetized and perfused with 4% paraformaldehyde. The right and left tibias were removed and will be submitted to mechanical tests and histological analysis. The X‐ray and bone densitometry results suggest that the 904nm laser diode is accelerating bone repair. The histological examination will help the understanding of the underlying mechanisms of this acceleration.
MYOCARDIAL BRIDGING ‐ A RISK FACTOR FOR MYOCARDIAL ISCHEMIA?
Authors: TEOFILOVSKI‐PARAPID Gordana , Jankovic Radmila M , Parapid Biljana J , Aleksandric Srdjan B , Nedeljkovic Ivana P , Beleslin Branko D , Simic Dragan V , Teofilovski Mirko C
Published: June 2021

Myocardial bridging (MBing) is a congenital anomaly the most frequently described over LAD. Recently, MBs are often related to myocardial ischemia (MI), left ventricle dysfunction, arrhythmias, and sudden cardiac death. The reported incidence of MBing on autopsies varies extensively (4.7%–86%), likely related to geographical regions, while national data have not yet been available.

Retrospective analysis of autopsy protocols of 721 consecutive autopsies (adults of both sexes) performed between July 2010 and June 2011 at the Institute of Pathology at FMUB, with LM studies (HE&Masson trichrome) of the tunneled vessel (TV) wall along with the morphometric analysis of the ventricular myocardium supplied by the TV (experimental group) and the ventricular myocardium of the same heart but with a different blood supply (control group) were done.

Myocardial bridges were described in 6/721 cases (0.8%): 5 were male and all over 70yrs. Only single MB were described over LAD. In all of the cases: a) TV had intensive atherosclerotic changes proximal to the MB and only focal in the tunneled part; b) there was intensive interstitial fibroses of the myocardium supplied by the TV.

We found strong evidence of MBing and MI, but do not consider the 0.8% being the true prevalence of MBing in Serbian population and suggested prospective studies. (This work was supported by MESRSerbia Grants: 41022;175030).
A Study of Facial Dimensions in Relation to the Gonial Angle in Cadaveric Specimens
Authors: Supple Elizabeth Lynn , Bruzzini Kristen Blake , Steinkoenig Colleen
Published: June 2021

It has been noted in some cases of cranio‐facial disformities that there is a relationship between facial height and the gonial angle of the mandible. This study focused on finding the relationship between these measurements in persons without any cranio‐facial disformities. Cadaveric specimens were used to determine a relationship between facial dimensions in the fixed facial bones and the gonial angle of the mandible. The distance between the anterior inferior edge of the nasal suture and the suture between the maxilla and the zygomatic bone was measured to determine the facial width. The facial height was measured from the mid‐sagittal alveolar process to the anterior inferior edge of the nasal suture. These measurements were compared to the gonial angle of the mandible to see what relationship exists.

Grant Funding Source: none
Anatomical variations in the superior thyroid artery in the South African population
Authors: Kolkenbeck‐Ruh Andrea , Hartmann Carol , Hutchinson Erin
Published: June 2021

For surgical disciplines, an understanding of the variations of the superior thyroid artery (STA) is essential as the STA is a crucial landmark during surgical procedures involving the thyroid gland. Therefore, the aim of this study was to investigate variations in the origin and branching pattern of the STA within a South African population. Further analyses included variations in the origin and branching pattern according to sex and symmetry.

Thirty‐two adult South African cadavers of European ancestry (18 males and 14 females) were dissected to expose the origin and branching pattern of the STA bilaterally. Observations were made on the origin and branching patterns of the vessel bilaterally. Frequency tables were used to determine the frequencies of the origin and branching patterns of the STA.

The STA originated most commonly from the external carotid artery at the carotid sinus (34.2%). This origin was observed more frequently on the left side in males than in females (p=0.04). In both sexes and for both left and right sides, the STA most commonly gave rise to anterior and posterior glandular branches.

South African populations of European descent exhibit similar variation in the origin and branching patterns of the superior thyroid artery to other European populations. Furthermore significant variation in the arterial supply to the thyroid gland exists between males and females. An understanding of these differences is important for successful thyroid surgery.
Body Mass Estimators in Fossorial Mammals and the Body Mass of Extinct Palaeanodonta (Pholidotamorpha)
Authors: Ahrens Heather E , Ruff Christopher B
Published: June 2021

Body mass is an important variable in many ecological, functional, and phylogenetic analyses. In order to test for differences between extant and extinct fossorial mammals, useful body mass proxies must be established. We tested multiple skeletal dimensions for their ability to estimate body mass in mammals that employ fossoriality, a specialized locomotor style. Given the derived morphology of the forelimb in those taxa, measures of the hind limb were expected to be more reliable than measures of the forelimb. Lengths, diaphyseal breadths, and articular breadths were taken on the humerus, ulna, and femur from eleven extant species representing several taxonomic orders, digging styles, and a range of body masses. Percent standard error of the estimate (%SEE) was used to assess the reliability of prediction equations. Length measures were the most reliable predictors of body mass; articular breadths of the femur were also reliable. Measures with a %SEE below 10% were used to estimate the body masses of several species of Palaeanodonta, which includes some of the most fossorial mammals of the Paleogene. The smallest measured palaeanodont, Epoicotherium unicum, is predicted to have weighed between 58.5 and 196.9 g, whereas the largest, Metacheiromys dasypus, is predicted to have weighed between 3.43 and 7.07 kg.

Grant Funding Source: None
Ontogeny of locomotion and head movement of the African spurred tortoise (Geochelone sulcata)
Authors: LeCheminant Ben Hamblin , Georgi Justin
Published: June 2021

The aim of this research is to describe the changes of head and neck kinematics as they relate to the growth of the African spurred tortoise (Geochelone sulcata). The basics of tortoise locomotion have been described previously. These studies, however, have not investigated locomotor changes in head kinematics due to body size. Infra‐red motion capture was used to obtain detailed kinematics of the head, limbs, and carapace on six live tortoises ranging in body mass from 10.8 to 66.2 kg. The results of the study showed that while there is no correlation between body size and stride duration (p = 0.198) or duty factor (p = 0.109), there is a significant positive correlation between size of tortoise and stride length (p = 0.016). To assess head stability, we compared the average magnitude of the maximum positive and negative differences between the rotational acceleration of the head and body in the sagittal plane. Head instability, regardless of body size, increases as stride average linear velocity increases (all p < 0.004). This rate of head instability increase with stride velocity was less marked in the larger tortoises compared to smaller tortoises (p = 0.017). Thus, larger tortoises maintain a more constant head stability. Our data do not differentiate increases in muscle mass, larger moment of inertia because of increased head mass, or other possible control factors as the source of this increase in head stability.
Atlatl saltation: A new model for richochetal locomotion
Authors: Naples Virginia L.
Published: June 2021

Small mammal saltatorial adaptations maximize escape in patchy environments where sudden positional changes are more effective than dashing away in one direction. Saltation is an independent series of longer jumps than body size predicts; saltators alter trajectory midleap using tails as rudders. Although from species differing in biogeographic range, phylogenetic origin and appearance in the fossil record, their ecological niches constrain saltators to show common morphologies. These include bipedal hopping, long, tufted tails, large eyes and auditory bullae, elongated feet and hind limbs with proximal muscle bellies. This study examines saltatory adaptations among forms of common appearance by comparing saltation to the atlatl, a device for increased human hunting efficiency that propels projectiles farther and more accurately. Such animals are atlatl saltators. The body parts determining these locomotor capabilities compare to those of an atlatl, but are analogous only when it is inverted. The atlatl saltator foot corresponds to the atlatl projectile and launcher, the calcaneus to the atlatl spur and the lower leg to the atlatl handle. The knee corresponds to the human wrist holding the device. Knee straightening equates to a forward wrist snap. In parallel, the animal uses large thigh muscles to move forward, while the human movement allows large trunk muscles to effect a corresponding atlatl movement.
In vitro biomechanical evaluation of fibular movement in below knee amputations
Authors: Asa Benjamin , Wilson Tim , Payne Mike , Dunning Cynthia , Burkhart Tim
Published: June 2021

Background

In below knee amputations (BKA), the tibia and fibula may be subjected to intrinsic and extrinsic forces resulting in fibular rotation and abduction, impeding ambulation. We investigated the effect of biceps femoris muscle tension and interosseous membrane (IOM) integrity on fibular kinematics in two BKA lengths. We hypothesize the stabilizing role of the IOM will be decreased in shorter amputations, and therefore, fibular movement will occur more readily.

Methods

Two cadaveric specimens, disarticulated at the knee, were amputated at 5 and 10cm lengths. Tibiae were mounted in a material testing machine and biceps femoris tendons (BFT) were sutured to the actuator. A cyclical loading pattern applied displacement‐dependent tensile forces (initial displacement of 3mm for 100 cycles @1Hz with increments of 1mm) to the BFT. The three‐dimensional kinematics of the tibia with respect to the fibula were measured. To evaluate the role of the IOM, the aforementioned procedure was repeated on specimens with a sectioned IOM.

Results

The 10cm BKA required more force than the 5cm BKA in the 100th cycle at each incremental displacement block, presumably due to decreased fibular movement. However, this difference is predicted to be negated when the IOM is sectioned.

Conclusion

Understanding the cause of fibular abduction in BKA will lead to recommendations for preventive surgical and rehabilitative measures.

Grant Funding Source: Departmental
Dimensions of the ankle ligaments: anatomical study and literature review
Authors: Duscher Dominik , Wenny Raphael , Weninger Patrick , Hirtler Lena
Published: June 2021

Introduction

With over 20,000 cases reported daily just in the United States damage of the ankle ligaments is one of the most frequent injuries of the musculosceletal system. Especially the lateral ligaments are prone to harm during supination movements. Depending on the severity of the injury conservative or operative therapeutic actions are indicated. For operative reconstruction precise anatomic information on the dimensions of the ankle ligaments is important and can help optimizing these procedures. The purpose of this study was to investigate the length and width dimension of the ankle ligaments and contrast the results with the published literature.

Material and Methods

17 non‐paired adult, formaline‐fixed cadaver ankles were dissected to expose the capsuloligamentous structures. After identification of the ligaments, dimensions were measured with a ruler and a sliding caliper.

Results

The dimensions of the ligaments were recorded. Means and standard deviation were calculated and discussed with the literature.

Conclusion

Studies of therapeutic options in severe ankle ligament injury have shown better results in anatomical reconstruction compared to other operative treatments. To optimize this procedures, exact anatomic information on the dimensions of the ankle ligaments should be beneficial.

Grant Funding Source: AAA
Intramuscular Innervation of Infraspinatus: A 3‐D Modeling Study
Authors: Hermenegildo Jason Aldeia , Ko Dominic M , Johnson Marjorie , Merrifield Peter A , Agur Anne M
Published: June 2021

Neuromuscular partitioning is important in defining functional differences within a muscle volume. It has been shown that individual neuromuscular partitions may be differentially affected in pathology. Neuromuscular partitions are defined by independent innervation and architectural differences. Infraspinatus (IS) is a functionally important rotator cuff muscle where neuromuscular partitioning has not been well studied. The purpose is to investigate the intramuscular innervation patterns of IS. In this pilot study, 5 formalin‐embalmed cadaveric have been dissected, digitized, and modeled to date. The suprascapular nerve (SSN) was digitized sequentially in short segments and the data was modeled using Autodesk® Maya®2012. The models were used to document the intramuscular innervation patterns/neuromuscular partitions. The SSN enters IS at the spinoglenoid notch as 1–3 nerves. In three specimens, one nerve entered the IS belly and divided into 3 main branches: (1) a superior branch (br) to superior part of the muscle belly; (2) a lateral br to upper inferior belly; (3) middle br to middle and lower inferior bellies. When SSN enters as 2–3 branches, a separate lateral and/or superior branch enter their respective regions. These results provide a detailed mapping of the intramuscular innervation of IS, providing evidence of neuromuscular partitioning within the IS muscle belly.
Regulation of early stages in clathrin mediated endocytosis revealed by quantitative analyses in living cells
Authors: Schmid Sandra L. , Aguet Francois , Antonescu Costin , Mettlen Marcel , Danuser Gaudenz
Published: June 2021

Clathrin‐mediated endocytosis (CME), which occurs at clathrin coated pits (CCPs), requires the major coat proteins, clathrin and AP2 complexes together with a myriad of endocytic accessory factors and lipids. CME proceeds through multiple stages, including initiation, stabilization of nascent CCPs, and maturation during which CCPs collect cargo molecules, grow in size and increasingly invaginate until they remain connected to the cell surface via a narrow neck. The GTPase dynamin assembles onto this neck and drives membrane fission to release clathrin coated vesicles and complete the process. Total internal reflection fluorescence microscopy (TIR‐FM) in living cells expression fluorescently‐tagged coat proteins is a powerful tool to study the assembly, maturation and dynamic behavior of CCPs; however, extracting reliable measurements from these TIR‐FM movies represents a major challenge. We have developed new methods for particle detection and tracking that enable accurate, unbiased and global analyses of CCP dynamics even at low signal:noise. These assays have revealed striking phenotypes resulting from the expression of mutant AP2 adaptors that could not be detected using conventional biochemical assays for CME.
Chondrogenesis in pellet cultures using multipotent mesenchymal stromal cells derived from Cd44−/− and wildtype mice
Authors: Veluci Roberta Medeiros , Knudson Warren , Knudson Cheryl B.
Published: June 2021

Murine multipotent mesenchymal stromal cells (mMSCs) are a promising model system to study regenerative cartilage biology. Either TGFβ or BMPs are used to induce MSCs to chondrogenesis or osteogenesis. Kartogenin (KGN) was recently shown to promote cartilage repair in osteoarthritic mouse models but its effects on the TGFβ and BMP pathways are unknown. Our previous studies showed that SMAD1, a factor in the canonical BMP pathway, interacts with the cytoplasmic domain of CD44, suggesting a mechanism whereby hyaluronan and its receptor CD44 may promote the cellular response to BMP and cartilage regeneration. Thus we tested the capacity of Cd44−/− and WT mMSCs to undergo chondrogenesis in pellet cultures containing BMP7 or TGFβ3 +/− KGN. We optimized a protocol for purifying mMSCs from tibial and femoral bone marrow cavities which yielded CD105+/CD11bmMSCs after 14d of culture (1.86% of original isolate). The osteogenic potential of mMSCs was detected with alizarin red. Chondrogenesis in pellet culture sections was evaluated using histology (e.g., Alcian blue/nuclear fast red, PNA) and morphometrics used to determine chondrogenic clusters. In terms of size, pellets treated with BMP7+KGN>; TGFβ3>; TGFβ3+KGN>; BMP7, and assessment of the chondrogenic clusters showed BMP7+KGN>; TGFβ3>; BMP7>; TGFβ3+KGN. Hence KGN may enhance BMP7 and hinder the TGFβ3 pathway to induce chondrogenesis. Supported by NIH R01‐AR39507.

Grant Funding Source: N/A
Repair the injured peripheral nerve by bridging neurons
Authors: Zhang chuansen
Published: June 2021

Restoration of the injured peripheral nerve with a long defect remains to be discussed now. We hypothesize that bridging neurons in the nerve defect can restore the injured peripheral nerve better. To confirm the hypothesis, experiments listed below have been performed. Firstly, isolation, identification and differentiation toward neurons and schwann cells of hair follicle neural crest stem cells (hfNCSCs) and bone marrow nerve tissue commited stem cells (bmNTCSCs) were investigated. we confirmed that hfNCSCs and bmNTCSCs can be induced to functional neurons. Secondly, we studied the development of the adult stem cell derived neurons in the microenvironment of the peripheral nerve and found that the cells could survive and keep the characteristics of the typical neurons. Thirdly, we laded the stem cell derived neuons on the allogenetic acelularized nerve grafts to bridge the sciatic nerve defect. We found that implanted stem cell derived neurons in vivo could survive for a long time, form the synapse and build the neural pathway. Reinervating time of effectors or receptors with neural tissue engineering nerve grafts transplantation was shorter than that with autologous nerve grafts and Schwann cells‐laden nerve grafts transplantation.
Going out on a limb: functional and developmental integration in human limb long bones
Authors: Auerbach Benjamin Miller , DeLeon Valerie Burke
Published: June 2021

Morphological integration (MI) describes the covariance of phenotypes, which in turn reveals information about shared genetic and developmental pathways underlying those phenotypes, as well as functional correlations. Previous research on MI in the elements of the limbs has focused on murine models and on comparisons among primates. The latter studies demonstrate an increase in the independence of long bone lengths among apes, especially humans. However, no research has examined integration among multiple dimensions—lengths, articulations, and diaphyses—in human limb bones.

This study examines external linear dimensions taken from lengths, articulations, and diaphyses of humeri, radii, femora and tibiae, using a global sample of 3625 human skeletons. Eigenvalue variances and covariance matrix comparisons were used to assess MI and modularity. Results show that similar dimensions (e.g., lengths) among homologous elements are more integrated than elements in the same limb. Lengths have the highest MI. There is independence between diaphyseal dimensions and lengths, corroborating previous findings. This indicates different sources of constraint on these morphologies from developmental and functional factors.
Classification of Sex and Ancestry from Metatarsal Bones by Radiographic Analysis
Authors: McCarthy Sarah A , Albano Michael , Nedresky Joseph , Cabo‐Perez Luis
Published: June 2021

It is difficult to accurately estimate sex and ancestry from incomplete skeletal remains; particularly when the skull and pelvis are absent. It is necessary to be able to accurately estimate a biological profile from multiple bones because skeletal remains are often found incomplete. Metatarsal bones are ideal candidates for estimation of sex and ancestry because they are more numerous than other post‐cranial bones and are usually found intact enclosed in shoes, protected from taphonomic influences. The goal of this study was to evaluate the presence of sexual and ancestral differences in metatarsal bones from a modern population. To evaluate these differences, we analyzed radiographs taken between 2003 and 2012, recording length and width measurements of metatarsal bones. We then performed discriminant function analysis to determine which measurements were most useful to classify sex and ancestry. We found that we could correctly classify sex and ancestry independently, 78.4% and 59.3% of the time, respectively; additionally 45.9% of individuals were correctly classified when sex and ancestry were pooled. In conclusion, our data suggests that there are sexual and ancestral differences present in metatarsal bones. Furthermore, our results suggest that a whole set of metatarsals is preferred to estimate sex and ancestry, with an emphasis placed on the first metatarsal bone.

Grant Funding Source: LECOM Internal Grant
Parallel lumbar and pelvic morphology in atelines and early hominids: clues to the earliest hominid adaptations to upright walking?
Authors: Machnicki Allison L , Haile‐Selassie Yohannes , Spurlock Linda , Mendes Sérgio L , Strier Karen B , McCollum Melanie A , Lovejoy C. Owen
Published: June 2021

Lumbar lordosis is a critical element of human bipedality, because it obviates the need for African ape style bent‐hip‐bent‐knee (BHBK) gait. Apes have eliminated lumbar mobility by trapping the most caudal lumbars between dorsally extended ilia. The earliest known hominids show the opposite state—emancipation of the most caudal lumbar by reduction in iliac height and expansion of sacral alar breadth. Ateline monkeys have flexible prehensile tails, which, when used simultaneously with their forelimbs for support, induce substantial lordosis. To determine if similar skeletal and ligamentous morphology underlies lumbar lordosis in atelines and hominids, we examined (dissection, MRI, X‐ray) the pelves and lumbar columns across anthropoids. We find substantial parallels between hominids and atelines including reduced iliac height and position of the sacrum relative to the ilium and pubis. These results indicate that lowering iliac height is a likely first step in directly evolving bipedality without any reliance on a BHBK gait.

Grant Funding Source: None
A quantitative assessment of the morphology of the piriform aperature as an indicator of race
Authors: Gardner Seth
Published: June 2021

A quantitative method for assessing the morphology of the piriform aperture was developed in order to investigate ancestry of an unknown skull. Discriminant function analysis predicted an unknown specimen's race with an accuracy of 77.4%. Step‐wise analysis produced an accuracy of 79.0%. The objective of this study was to quantitatively assess piriform aperture morphology as a tool for discriminating ancestry between Black and White populations. Twenty‐four linear measurements as well as the perimeter of the anterior nasal aperture were taken from digital images using the Adobe Acrobat Pro 9.0 Measurement Tool. Multivariate analysis, specifically Discriminant Function Analysis (DFA), was used to analyze the data. Eleven of the twenty‐five variables measured in this study proved to be racially dimorphic. A multivariate discriminant function produced a useful equation to predict an unknown specimen's race based on the discriminate function score with an accuracy of 77.4%. Step‐wise discriminant function analysis, using only the three most significant variables, produced an accuracy of 79.0%. Where the sex was known to be male, DFA generated a function using only one variable (105° from centroid) to accurately discriminate between the two races. The results of this study bear out the hypothesis that one can determine the race or ancestry of an unknown skull, using the Adobe Acrobat Pro 9.0 Measurement Tool.

This research was funded by Palmer College. This investigation was conducted in a facility constructed with support from Research Facilities Improvement Program Grant Number C06 RR15433–01 from the National Center for Research Resources, National Institute of Health.
Morphological integration of the human mandible in two pre‐ Columbian skeletal samples
Authors: Reeves Nicole M. , Wheat Amber , Dudzik Beatrix
Published: June 2021

The human skull is a complex structure composed of many semi‐independent but integrated units. Functionally, the cranium is often partitioned into three modules: the vault, basicranium, and splanchnocranium. While the mandible is an integral functional component of the skull, it is rarely examined in the field of human morphological integration. In this study, we investigate patterns of integration between the vault, basicranium, and mandible in adult skulls from two related pre‐Columbian populations—Caddo and Arikara—using 3D geometric morphometrics. The Caddo exhibit artificial cranial deformation (ACD), while the Arikara are nondeformed (ND). This study tests two null hypotheses: (1) ACD skulls will demonstrate the same levels of overall integration as ND skulls, and (2) patterns of integration between the mandible and cranial base are not affected by ACD. Procrustes superimposition is used to scale and rotate the coordinate data, and principal component analysis and discriminant function analysis are used to assess shape differences and variation between the samples. Results indicate that integrated shape changes are complex, but the overall pattern is similar among the ACD and ND samples. This is one of the first studies to examine how patterns of integration between the mandibule and other modules of the cranium are affected by ACD.

Grant Funding Source: AAA
Inhibition of Homologous Recombination and the DNA damage Checkpoint by Caffeine
Authors: Tsabar Michael , Eapen Vinay , Memisoglu Gonen , Haber James E.
Published: June 2021

In response to DNA double stranded break H2AX undergoes rapid phosphorylation by the checkpoint PI3 Kinases ATM and ATR. This modification, termed γ‐H2AX, persists as long as the damage is present. We used caffeine, a known PI3 kinase inhibitor, to ask if the modification requires continuous activation of ATM and ATR in Saccharomyces cerevisiae. We find that the inhibition of Mec1 and Tel1, yeast's ATR and ATM homologues respectively, does not lead to the loss of γ‐H2AX. Also we show, for the first time, that upon caffeine treatment Rad53 and Rad9 are rapidly dephosphorylated, and that this is reversible when caffeine is removed from the media. Lastly we show that caffeine inhibits homologous recombination by eliminating resection and evicting Rad51 filaments from the DNA in a manner independent of Mec1 and Tel1 inhibition. These results demonstrate two independent pathways by which caffeine treatment alters the DNA damage response.
Ionizing radiation resistance mediated by the novel selenium‐containing antioxidant selenoneine
Authors: Imamura Shintaro , Yabu Takeshi , Ishihara Kenji , Yamashita Yumiko , Yamashita Michiaki
Published: June 2021

The novel selenium (Se)‐containing imidazole compound, 2‐selenyl‐trimethyl‐histidine, selenoneine, has been identified from the blood and tissues of the tuna (Yamashita et al, JBC, 285, 18134–18138, 2010). This compound had a strong radical scavenging activity, and had a protective function under oxidative stress conditions. In order to elucidate the molecular mechanism of DNA repair by Se, DNA damage response to γ‐irradiation was characterized in zebrafish embryos (8 hpf) in the presence of selenoneine at 10–100 nM in culture medium at 0.1–8 Gy. Although apoptosis was induced and ionizing radiation had no discernable effects on overall growth and morphology, TUNEL‐positive apoptotic cells were detected in brain, eye and spinal cord. Such apoptosis in CNS was reduced by the treatment of selenoneine at 25 nM after γ‐irradiation. CDC48/VCP/p97, which promotes both ubiquitin‐proteasome system and autophagy, was phosphorylated at Ser‐784 in response to DNA damage, and the treatment of selenoneine blocked CDC48 phosphorylation, suggesting that DNA double strand break by ionizing radiation might be reduced by selenoneine radical scavenging function. Knockdown of CDC48 by antisense morpholino oligos enhanced abnormal morphogenesis and apoptosis in CNS by γ‐irradiation. Therefore, Se redox mechanism by selenoneine and CDC48‐mediated autophagy had a protective function against ionizing radiation.
Cloning and expression of uvrA gene of Corynebacterium pseudotuberculosis to value its role in NER pathway
Authors: Castro Bárbara Catarina Teodoro , Resende Bruno Carvalho , Araújo Priscila Assis , Machado Carlos Renato , Azevedo Vasco Ariston Carvalho , Miyoshi Anderson , Lopes Débora Oliveira
Published: June 2021

Caseous lymphadenitis is a infection that affects sheep and goats and caused by Corynebacterium pseudotuberculosis. DNA repair, the target of this study, is essential for survival of any organism, to prevent potential deleterious effects. The objective of this study was to characterize uvrA gene from C.psudotuberculosis using in silico and in vitro techniques. In silico analyzes were performed to check the physical and chemical parameters, search for conserved domains and modeling of 3D structure using bioinformatics programs. The analysis shows that the protein has a molecular weight of about 101.5 kDa and a conserved domain of ABC transporters family and ABC excinuclease. The amplification of uvrA gene was carried out using a Long Range PCR Kit and the insert was cloned into the TOPO PCR 2.1 vector. E. coli strain DH5α was electroporated with recombinant plasmid and after that these were digested with restriction enzymes BamH1 and Xho1 for subsequent cloning into the expression vector pProEX HTB. The constructions were confirmed by sequencing. Financial support: CNPq and FAPEMIG
TRF1 interacts with XRCC4 and XLF at telomeres
Authors: Alsina Katherina Maria , Nunez Tatiana , Abbas Zamzam , Berhe Feven Bereket , Le Timothy Le Christian , Sen Pramila , Jain Renu , Jabbur James Ramzi , Ribes‐Zamora Albert
Published: June 2021

At telomeres, the shelterin complex is entrusted with a protective role against DNA repair mechanisms that could mistakenly recognize the natural ends of chromosomes as the product of a double strand break, and create telomeric end‐to‐end fusions that can drive genome instability and tumorigenesis. Paradoxically, telomeres also harbor several members of the DNA repair machinery whose actions they are trying to block, like members of the NHEJ pathway. How these NHEJ proteins are prevented from engaging in NHEJ at telomeres is not fully understood. Using protein fragment complementation assay, we have detected novel interactions between shelterin proteins TRF1 and TRF2 with XRCC4 and XLF, two NHEJ proteins that interact and are involved in later steps during DSB repair. Furthermore, cells expressing interacting TRF1 and XRCC4 proteins displayed clear punctate staining that co‐localized with other telomeric proteins, indicating that XRCC4 and TRF1 can interact at telomeres. Mutations that disrupt XRCC4‐XLF interaction or known phosphorylation sites for ATM and DNA‐PKcs kinases did not impair XRCC4‐TRF1 interaction nor did they impair XRCC4 localization at telomeres. Overall our results suggest that NHEJ proteins XRCC4 and XLF can be recruited to telomeres through interactions with shelterin proteins TRF1 and TRF2 in ways that require neither XRCC4‐XLF interaction nor ATM or DNA‐PKcs phosphorylation.
Systems Biology Analysis of HIV‐Host Interactions
Authors: Young John A.T.
Published: June 2021

Viruses like HIV‐1, the cause of AIDS must exploit numerous cellular functions to facilitate their replication. At the same time they must counteract both innate and adaptive host immune responses to avoid virus clearance. Along with our collaborators we are using systems biology approaches to dissect the networks of cellular cofactors and restriction factors that regulate virus replication. These studies have revealed a novel role for the cellular sulfonation pathway in regulating HIV‐1 gene expression, numerous novel cellular substrates for HIV‐1 accessory factors, and has revealed a new role for viral apoptotic mimicry in regulating the cellular antiviral response. Our progress in these areas will be discussed.
In Vivo role of Site‐Directed Cleavage of DNA by the RecA‐dependent nuclease Ref
Authors: Baker Staci J. , Ronayne Erin A. , Cox Michael M.
Published: June 2021

The Ref protein, encoded by bacteriophage P1, is a RecA‐dependent nuclease. A member of the HNH nuclease family, Ref will cleave DNA only if it is bound by the RecA protein. When RecA filaments are formed on an oligonucleotide and used to form a D‐loop in a target duplex DNA, Ref will cleave both strands of the target DNA within the D‐loop. In this mode of action, Ref is effectively a universal restriction enzyme. The function of Ref nuclease in the life cycle of bacteriophage P1 and related bacteriophages is a mystery. In this report, we will present data related to the in vivo function of Ref. In particular, wild type Ref protein both enhances certain types of RecA‐dependent recombination events and also suppresses P1 lysogenation.