bluuebunny/crossref_metadata_2025_split huggingface dataset:
Angiogenic Gene Modification Of Skeletal Muscle Cells To Compensate For Age‐related Decline In Function
Authors: Zhan Wang , Delo Dawn , Eberli Daniel , Atala Anthony , Soker Shay 
Published: June 2021

INTRODUCTIONIn this study, we investigated muscle regenerative ability in different age groups of experimental animals.METHODMuscle precursor cells (MPCs) were cultured from mice. In vitro analysis included cell growth assay, quantitative myotube formation. In vivo studies involved cells transfected or not transfected by adenovirus encoding Vascular Endothelial Growth Factor (VEGF). MPCs were injected subcutaneously into mice. After 2 and 4 weeks, the engineered muscle samples were harvested and analyzed.RESULTSThe young mice MPC growth rate and myotube formation were greater than mature mice and old mice MPC. Bioengineered muscle tissues were smaller and less contractile in old mice group. With expressing recombinant VEGF, bioengineered muscles in all age groups had a greater mass, increased neovascularization, and contractile function. Histological analysis of the in vivo engineered muscle tissue stained positive for muscle specific proteins.CONCLUSIONSThis study demonstrates the MPC regenerative possibilities in young and old mice, and augmentation of neovascularization increased muscle mass and strength of contractility of engineered muscle tissues. Funding NIH Ruth L. Kirschstein National Research Service Award Individual Fellowship (F31), the Novartis‐Stiftung, the Roche Research Foundation, GEBERT RÜF STIFTUNG, and CROWN Foundation.
The Bilateral Nature and Origin of the Medial Circumflex Femoral Artery
Authors: Beckman Michael , Frumberg David , Márquez Samuel 
Published: June 2021

The medial circumflex femoral artery (MCFA) largely supplies the posterior aspect of the femoral head while also contributing to the neck of the femur. It thus plays a major role in the healing of hip fractures and prevention of subsequent avascular necrosis in this susceptible area of the human body. The MCFA is commonly described in anatomy and orthopaedic textbooks as typically arising from the deep artery of the thigh close to the femoral artery. In this study, 76 medial circumflex femoral arteries from 38 adult cadavers representing an urban population were surveyed during the first year gross anatomy courses for College of Health Related Professions and College of Medicine students at SUNY Downstate Medical Center. One cadaver was excluded due to a synthetic aortic‐iliac‐femoral arterial system yielding a final sample size of 74 hips from 37 cadavers. Results show a large amount of variation in the origin of the MCFA with the following four general patterns emerging from this study: 1) MCFA branches off the deep artery of the thigh; 2) MCFA branches off the femoral artery proximal to the origin of the deep femoral vessel; 3) MCFA branches off the femoral artery distal to the origin of deep femoral vessel; and 4) MCFA branches off at the bifurcation of the femoral and deep arteries of the thigh in a "trifurcation" pattern. The low statistical finding of only 37.8% of our sample with the MCFA branching off the deep femoral artery is surprising given the high frequency reported in the literature. The fact that about 50% of cadavers from this study showed bilateral asymmetry emphasizes not only the highly variable nature of MCFA branching patterns but decreases the predictive power of symmetry in any given individual.Grant Funding SourceDo Not Have Funding
Clavicular length asymmetries in primates indicative of handedness in daily activity
Authors: Grisotti Gabriella , Gannon Patrick J. , Marquez Samuel 
Published: June 2021

Handedness is well understood in humans but not for other primates. Some studies in non‐human primates have focused on behaviors such as reaching for food correlated with hemispheric specialization. Other studies reported asymmetry in upper limb bones of Pan troglodytes to support handedness. Given relations of the upper limb and clavicle, we hypothesized that evidence of handedness would be manifest in measurements of clavicular length. We collected data from chimpanzees (n=8), rhesus (n=2), and spider (n=2) monkeys to represent great apes and Old and New World monkeys from the Division of Mammalogy at the American Museum of Natural History. Preliminary results showed a non‐significant but 62% rightward asymmetry in chimpanzees but essentially symmetry in the monkeys. Clearly, a larger sample size is required to explore these findings further. The relationship of the clavicle between the axial and appendicular skeletons means that it is responsible for transmitting a great deal of force. Activities of the handed upper limb may well have a differential effect on the clavicle. Thus, the clavicle may offer a unique approach to recognize handedness even in extinct primates.Grant Funding SourceDo not have funding
An anatomical exploration of femoral angle of inclination in multi‐regional populations: clinical implications
Authors: Charubhumi Daniel , Schachar Jeffrey S , Chaudhri Ali , Márquez Samuel 
Published: June 2021

The angle of inclination (AOI) is clinically relevant because of a predisposition for congenital femoral dysplasia. While AOI of the femur varies with age, gender, and ontogenetic growth patterns, abnormal AOI conditions increase stress at the hip joint and significantly affect gait. This study examined AOI in three ethnic/racial populations from European, African, and Asian ancestry. An Inuit group dated 500 BCE ‐ 500 CE was sampled from archaeological remains of Ipiutak populations from the osteological collections of the American Museum of Natural History. To measure AOI, a line was drawn from the midpoint of the intercondylar fossa to the greater trochanter projecting proximally through the shaft of the femur, and a second line was drawn from the femoral head to the femoral shaft through the center of the femoral neck; the AOI was defined by the intersection of these two lines. Preliminary results showed significant AOI differences (p < 0.01) found only between the Inuit and European populations. Developmentally, the AOI is widest at infancy and gradually narrows, ending its decline in adulthood. Inuit femurs presented with the greatest AOI of 139 degrees, demonstrating the orthopaedic condition of coxa valga. The fact that these findings suggest a possible increased prevalence of congenital femoral dysplasia in these populations has considerable clinical implications and will require further study.Grant Funding SourceN/A
Femoral neck anteversion: an orthopaedic evaluation between multi‐regional populations
Authors: Schachar Jeffrey Scott , Charubhumi Daniel , Marquez Samuel 
Published: June 2021

Femoral neck anteversion (FNA) varies with age, sex, and ontogenetic growth pattern. This study examined African (Af), European (E), and two Asian groups, one of which, an Inuit (I) population from osteological collections of the American Museum of Natural History. FNA is the angular difference between proximal and distal ends of the bone. A line was drawn in the coronal plane through the lateral and medial femoral condyles. A second line was drawn from the greater trochanter to the head of the femur, through the center of the femoral neck. The intersection forms the anteversion angle. Selecting only male and right femurs controlled for the confounding variables of sex and asymmetries. Statistically significant differences (p < 0.05) were found between E and I groups and between Af and I groups. Average anteversion angle for the Af group was 15.7 degrees, 11.1 for the E group, 33.4 for the I group, and 23.0 for the Asian group. Developmentally, the amount of FNA is 40 degrees at birth, decreasing to ~15 degrees at maturity. The study revealed that the I sample presented with the largest angle, displaying excessive femoral torsion. Femoral torsion is viewed symptomatically as in‐toeing, which usually corrects without treatment as the child matures. Our results suggest that the Inuit sample may not have undergone the physiologic torsion reduction of 25 degrees as seen in the other representative samples.Grant Funding SourceN/A
Forelimb anatomy of the aardvark, Orycteropus afer (Mammalia, Tubulidentata)
Authors: Voegele Georgina McKusick 
Published: June 2021

Soft tissue anatomy, especially myology, has been largely neglected by systematic biologists reconstructing the phylogeny of Mammalia. This is due to inadequate description and illustration of structures in many mammalian orders, and conflicting or inaccurate terminology in the existing descriptions. While anatomical terminology has been standardized for the domestic animals and their myology is well known, many rare mammals have not been dissected in decades and the available descriptions lack standardization and often compare taxa now known to be genetically unrelated. This leaves systematic biologists unable to verify muscle character states in the exotic mammals whose evolutionary relationships are most controversial. For example, little morphological evidence has been found to support the clade Afrotheria, which unites several mammalian orders thought to be unrelated until the advent of genetic sequencing.Here I present the forelimb anatomy of the aardvark, Orycteropus afer, the one living member of the order Tubulidentata and a member of the clade Afrotheria, based on my dissections of two alcohol‐preserved museum specimens. These dissections are part of a dissertation project to document the limb anatomy of several mammalian orders, particularly the little known smaller afrotheres, for use in systematics research. This research was supported by the Field Museum of Natural History Visiting Scholar Fund.Grant Funding SourceNational Science Foundation
Statistical analysis and mathematical modeling of collective cell motion in endothelial and epithelial cultures
Authors: Szabo Andras , Unnep Renata , Czirok Andras 
Published: June 2021

Cell migration within confluent monolayers is instrumental in several morphogenic processes. The phenomenon was studied with time‐lapse microscopy of in vitro monolayer cultures of endothelial and epithelial cells. Statistical analysis of cell trajectories reveals that motile confluent cultures may behave either as vortical fluids or as deforming elastic sheets. The type of behavior is determined by the duration of the cell‐cell contacts: trajectory analysis of adjacent cell pairs reveals that endothelial cells diverge easily, while other epithelial cells adhere to each other for an extended time period. The observed flow fields can be explained by our generalized cellular Potts model; in the model we represent cell polarization and the resulting self‐propulsion, as well as links between the cytoskeleton of adjacent cells.The work was funded by the NIH (R01 HL87136), and the Hungarian National Office for Research and Technology (NKTH).
A method to quantify apoptosis in mouse embryos using flow cytometry
Authors: Mallela Murali K , Gutierrez Juan Claudio , Hrubec Terry C 
Published: June 2021

Cell proliferation, differentiation and apoptosis play a major role in morphogenesis and organogenesis of the developing embryo. Teratogens can cause alteration in apoptotic patterns in the developing embryo which may lead to fetal malformations; however, no studies have quantified apoptosis in early embryos. We developed a protocol to quantify apoptosis in CD‐1 mouse embryos using flow cytometry. Valproic acid (VA) and diabetes mellitus were then used to induce neural tube defects and cardiac malformations respectively to evaluate apoptotic changes induced by known teratogens. Apoptotic levels were quantified in head regions of VA exposed embryos and hearts of hyperglycemic fetuses. Embryonic cells were disassociated enzymatically followed by differential staining with Annexin‐V and propidium iodide to differentiate apoptotic, dead and live cells. Apoptosis levels increased in the head regions of VA exposed embryos (18.1± 2.2 %) than the controls (12.7 ± 1.9 %). Increased levels were also observed in the hearts of hyperglycemic fetuses compared to controls. TUNEL assay, used to localize apoptotic regions, showed increased apoptosis along the open neural folds of the VA exposed embryos. This is the first report using flow cytometry to evaluate apoptosis in early embryos, and the results indicate this may be a useful method to quantify relative changes in apoptotic levels in teratogen exposure.Grant Funding SourceNIH grant K01RR16241‐01e
N‐cadherin and actin are present at desmosomes in the seminiferous epithelium of the adult testis
Authors: de Asis Marc Aristaeus Kaw , Young J'Nelle S. , Vogl A. Wayne 
Published: June 2021

Desmosomes in the seminiferous epithelium of the adult mammalian testis are not as well defined morphologically as those in most other epithelia. Moreover, the plasma membrane related plaques in Sertoli cells are linked to vimentin filaments, not to keratin filaments. Previous studies have indicated that these desmosomes contain classical actin‐related cadherins, thereby raising the interesting possibility that these junctions may have characteristics both of actin and of intermediate filament based junctions. To explore this possibility, we probed the seminiferous epithelium of adult male Spraque Dawly rats for N‐cadherin, actin, and plectin. In fixed frozen sections, N‐cadherin labeled sites at the base of the epithelium that only partly appeared to colocalized with the intensely labeled actin in ectoplasmic specializations. In fragmented epithelium, the actin probe reacted with plectin positive desmosomes. At the ultrastructural level, the probe for N‐cadherin clearly labeled desmosomes. Significantly, an antibody to actin also reacted above background with desmosomes and was localized close to the cytoplasmic face of the membrane related plaques. Our results are consistent with the conclusion that desmosomes in the seminiferous epithelium have characteristics both of actin and of intermediate filament based attachment junctions.Grant Funding SourceNSERC
Endothelial mechanosensing by primary cilia
Authors: Hierck Beerend P , Van der Heiden Kim , Egorova Anastasia D , Van de Pas Simone , Poelmann Robert E 
Published: June 2021

Primary cilia are microtubule containing membrane protrusions which are associated with fluid sensing in endothelial cells (EC). We show that ciliated EC are present at atherosclerotic predilection sites in adult mice under athero‐prone flow conditions, but not in athero‐protected areas. Upon plaque formation in Apoe−/− mice, as well as after experimental induction of flow disturbance, the number of ciliated EC is increased. The aim of this study was to relate the presence of a primary cilium on EC to the flow profile to which these cells are exposed to, in vitro and in vivo, and to decipher its role in the mechanosensor for shear stress. We analyzed EC under steady, pulsatile and oscillatory flow and show that ciliation is indeed induced by oscillatory fluid flow. EC respond to shear with an acute intracellular calcium transient, and with a prolonged change in gene expression. The latter is mediated through the microtubular part of the cytoskeleton to which the primary cilium is connected. Experimental de‐ciliation of EC largely abolishes the shear response, which shows that the primary cilium sensitizes EC for fluid shear stress.The mechanosensor for shear forces facilitates a two step response in endothelial cells: (1) an acute and transient increase in calcium and release of stored vasoactive substances, and (2) a prolonged phenotypic adaptation which is reflected by altered morphology and gene expression.
Analysis of Tissue Layer Stratification in Cells over Expressing Cdk4 and Long‐time Cultured Cells with Green Fluorescence Protein
Authors: Terefe Eyuel Solomon , Vaughan Melville Bailey 
Published: June 2021

I examined whether the stratification of keratinocytes, the primary cells of skin epithelium, that over expressed the Cdk4 and telomerase (hTER)‐which made them into immortal cells, had an influence on whether the keratinocytes tissue layer was organized as normal keratinocytes. These immortal keratinocytes were engineered to overexpress GFP so they could be used in wound healing models to monitor keratinocytes migration in real time as they healed a wound. The goal was to determine if GFP affected their ability to stratify and differentiate similar to normal keratinocytes. I arrived at my results by examining immunoflourcense stains for the presence of proteins within the epithelial layers/ dermal compartment in their proper location and orientation. The staining was for the proteins: Involucrin, Keratin 14, and basement membrane proteins collagen IV and laminin‐5. I found that the organization of the epithelial layers of the over expressing keratinocytes to be similar as those of non‐over expressing cells, which means that keratinocytes maintained epithelial cellular organization although they were going through abnormal proliferation. I also analyzed the epithelium layer organization of keratinocytes that were cultured for prolonged period. I also found that skin equivalents that were maintained for long period of time were similar to those that weren't maintained for such a long period.Grant Funding SourceLS‐OKAMP
Atypical PKC isoforms are expressed in regions of interest to respiratory motor control
Authors: Guenther Courtney H. , Baker‐Herman Tracy L. , Windelborn James A. , Mitchell Gordon S. 
Published: June 2021

Atypical protein kinase C (PKC) isoforms play important roles in many neural processes, including synaptic plasticity and neurodegenerative disease. However, the role of atypical PKCs in respiratory motor control has not been explored. Since preliminary data from our laboratory suggest that atypical PKC inhibition alters respiratory nerve activity in anesthetized rats, we explored the neuroanatomical distribution of atypical PKCs in brainstem and spinal cord regions known to play critical roles in respiratory control. Specific areas examined include neurons associated with: rhythm generation (preBötzinger Complex), pattern formation (Bötzinger Complex, ventral respiratory group), chemosensitivity (retrotrapezoid nucleus, nucleus tractus solitarius), and respiratory motor nuclei (phrenic and hypoglossal). Brainstem and spinal cords from perfused Lewis rats were sectioned (40μm slices) and stained with an antibody targeting the C‐terminus of atypical PKCs. Results: atypical PKCs are expressed in phrenic and hypoglossal motoneurons, nucleus tractus solitarius, and the Bötzinger complex. Identifying the neuroanatomical distribution of atypical PKCs in regions critical for respiratory motor control is a first step towards understanding their potential role in the control of breathing (Supported by NIH HL 80209).Grant Funding SourceNIH HL 80209
The Origin of Inputs to the Monkey Central Mesencephalic Reticular Formation (cMRF)
Authors: Warren Susan , Zhou Lan , May Paul J. 
Published: June 2021

The central mesencephalic reticular formation (cMRF) is a midbrain region reciprocally connected to the superior colliculus (SC) where stimulation elicits horizontal saccades. Here we sought to identify its sources of input, by injecting retrograde tracers into the cMRF of M. fascicularis monkeys. In the diencephalon, labeled neurons were observed in the zona incerta, hypothalamus and the ventral lateral geniculate, ipsilaterally. The main source of midbrain input to the cMRF outside the SC was the ipsilateral substantia nigra pars reticulata, and contralateral cMRF. Within the pons, the paramedian reticular formation displayed a few cells, mostly ipsilaterally. Labeled neurons were found bilaterally in the nucleus prepositus hypoglossi and in the interposed and fastigial nuclei. A few neurons were present at medullary levels. It has been suggested that the cMRF might provide a conduit to the SC for ascending information on saccade duration and direction. While some feedback was observed for most of the cMRF's downstream targets, extensive connections to support this proposed function were not found. Instead, the cMRF seems to receive its greatest input from motor, as opposed to sensory, structures that also supply the SC. If the cMRF reticulotectal projection is inhibitory, then the GABAergic cells in the nigra and zona incerta are in position to release this inhibition via the cMRF.Support: NIH Grant EY014263
The Macaque Midbrain Reticular Formation Sends Different Types of Feedback to Each Side of the Superior Colliculus
Authors: Wang Niping , Warren Susan , May Paul J. 
Published: June 2021

The central mesencephalic reticular formation (cMRF) likely plays a role in gaze control, as cMRF neurons receive tectal input and provide a bilateral projection back to the superior colliculus (SC). Whether this feedback is excitatory or inhibitory is unknown. In this study, biotinylated dextran amine (BDA) was injected into the cMRF of M. fascicularis monkeys to anterogradely label reticulotectal terminals and retrogradely label tectoreticular neurons. BDA labeled profiles in the ipsi‐ and contralateral intermediate gray layer (SGI) were examined electron microscopically. Postembedding GABA immunochemistry was used to identify putative inhibitory profiles. Nearly all (98%) of the ipsilateral BDA labeled terminals were GABA‐positive, but profiles postsynaptic to these terminals were exclusively GABA‐negative. Most (81 %) of the retrogradely labeled tectoreticular profiles in ipsilateral SGI were GABA−. In addition, BDA labeled terminals were observed contacting BDA labeled dendrites, indicating the presence of a monosynaptic feedback loop connecting the cMRF and ipsilateral SC. In contrast, within contralateral SGI, 54% of the BDA labeled terminals were GABA+, while 46% were GABA−. Most of the postsynaptic profiles were again GABA−. These results indicate the cMRF inhibits the side of the SC that activates it during a saccade, but it has more complex effects on the contralateral SC.Support EY014263
Growth factor and cytokine regulation in ventral cervical spinal cord following C2 hemisection in rats
Authors: Windelborn James A , Mitchell Gordon S 
Published: June 2021

Large shifts in growth factor and cytokine expression have been reported in several models of spinal injury. However, published reports seldom specify changes in spinal motor areas of the ventral horn, particularly in segments below the injury. Understanding injury‐induced changes in these molecules will help define abnormalities in the molecular environment that underlie spontaneous functional recovery and/or alter the response to therapeutic intervention. Thus, we studied changes in the expression of growth factors and inflammatory mediators in the spinal ventral horn after C2 hemisection (C2HS) or sham surgery in male Lewis rats 1‐28 days post‐injury. Relative changes in BDNF, VEGFA, TNF‐α and TrkB protein levels at the site of injury and near the phrenic motor nucleus (C4) were quantified by densitometry and expressed as a percentage of time‐matched sham rats. Surprisingly, no protein measured was changed 14 or 28 days post‐C2HS. BDNF levels were increased in C2 ventral horn 1 and 3 days post‐C2HS (p<0.01), but not in C4. TNF‐α increased 1 day post‐C2HS in both C2 and C4 (p<0.05), but was only marginally elevated 3 days post‐C2HS (p=0.058). Thus, despite traumatic spinal injury, BDNF and TNF‐α expression only transiently increase in the cervical ventral horn whereas other growth factors/receptors studied are unaffected by injury.Support by NIH HL69064 and HL007654 (JW)Grant Funding SourceNIH HL007654