bluuebunny/crossref_metadata_2025 huggingface dataset:
Treatment with Momordica charantia Fractions Disrupts Energy Metabolism and Elevated Reactive Oxygen Species’ Generation in Human Lung and Breast Cancer Cell Lines
Authors: Ehigie Adeola Folasade , Wei Peng , Wei Taotao , Yan Xiyun , Ojeniyi Fiyinfoluwa Demilade , Ehigie Leonard Ona
Published: April 2020

The mechanism of action of mitochondrial targeted anti‐cancer drugs relies on their ability to disrupt the energy producing systems of cancer cell mitochondria. Momordica charantia (MC), an acclaimed anticancer plant in folk medicine, is here investigated for the probable mechanism by which it kills cancer cells. The ethyl acetate (D3), n‐hexane (D4) and dichloromethane (D5) fractions were obtained from the Crude Water‐Soluble extract (CWSE) of MC via solvent partitioning and then administered on MDA‐MB 436 and A549 cells at 100 and 125 μg/mL for 24 h. Cytofluorometric techniques, using TMRM and H2DCF‐DA as fluorescent probes were employed to assess the effect of MC on mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) generation respectively, while ATP levels were measured using the luciferin‐luciferase reaction. Mean respiring cell numbers obtained from Dot graphs and Histograms were further analyzed, yielding significant (P<0.05) MMP depressions only at 125μg/mL in both MDA‐MB 436 (43.08±2.33) and A549 (24.84±2.51) cells treated with D3 in comparison with untreated cells (175.8±6.13 and 131.7±11.21 respectively). In the same vein, only MDA‐MB 436 cells treated with D3 and D5 at 100 and 125 μg/mL significantly elevated ROS levels in comparison with untreated (100.0±2.16) cells; the highest generation being 332.1±3.20, seen in D3‐treated cells at 125 μg/mL. Furthermore, all fractions (D3‐D5) tested on A549 cells at 100μg/mL and 125μg/mL generated significantly higher levels of ROS compared with the untreated (42±4.50) group. The highest observed levels of generated ROS were seen at 125μg/mL in the D5‐treated cells (536±4.00). For intracellular ATP generation, significant (P<0.05) depletion was observed in both MDA‐MB 436 cells [only at 125μg/mL of D3 (80.63±4.70) compared with the control group (106.7±5.69)] and A549 cells [D3 at 125 μg/mL (43.68±0.13) < D5 at 125 μg/mL (52.56±4.16) < D3 at 100 μg/mL (59.84±5.39) compared with the control group (98.57±0.66)]. On the whole, this study suggests that the tested fractions of MC induced a mitochondrial‐dependent cell death in selected cancer cell lines.

Support or Funding Information

Organization for Women in Science for the Developing World (OWSD) and Swedish International Development Cooperation Agency (SIDA)
Tamarindus indica Fruit Pulp Restores Reproductive Function in Sodium Fluoride Administered Rats
Authors: Ajani Emmanuel Oladipo , Usman Lutfat Abimbola
Published: April 2020

Background

Infertility is often associated with increased depression and stigmatization of infertile individuals. Although there exists in Africa, fertility inducing herbal condiments, however the pharmacology of most of these agents are unknown.

Objective

This study investigated the prophylactic and ameliorative effect of ethanolic fruit pulp extract of Tamarindus indica on sodium fluoride induced reproductive dysfunction.

Methodology

T. indica fruit pulp was extracted in ethanol by maceration. The filtrate was thereafter oven dried at 60°C. Seventy rats of both sexes weighing 120 ± 5 g randomized into 7 groups of 10 rats each were used for the study. Group 1 was placed on placebo. Group 2 and 3 were first placed on 200 and 400 mg/kg of the extract respectively for 4 weeks followed by concomitant administration 20 mg/kg NaF. Groups 4, 5 and 6 were initially administered with 20 mg/kg NaF for 45 days followed by 4 weeks’ administration of 200 and 400 mg/kg T. indica (4 and 5) and 50 mg/kg clomid (group 6).

Result

The sperm function indices showed that administration of 400 mg/kg extract significantly increased sperm count and restore the sperm motility, morphology and viability to values not different from the control. Follicle stimulating hormone and progesterone also compared favorably with the control. The leutinizing hormone, testosterone, and oestrogen at 400 mg/kg dose were significantly raised. The histopathological analysis of the testes of rats pre‐etreated with the extract showed the presence of large interstitial space in the seminiferous tubules while a relatively normal histomorphological representation was seen in rats treated with the extract after sexual dysfunction induction.

Conclusion

Our results indicate that administration of the fruit pulp extract of Tamarindus indica at a dose of 400 mg/kg body weight is efficacious for the management of infertility.

Support or Funding Information

Personall
IN VITRO EVALUATION OF THE CYTOTOXIC POTENTIAL OF Securidaca longepedunculata ON HUMAN BREAST ADENOCARCINOMA (MCF‐7) CELLS
Authors: Minari Joseph , Oyedele Olaoye , Okpuzor Joy , Aja Joyce , Adewole Richard
Published: April 2020

The prevalence of cancer has increased interest in the role of complementary and alternative medicine, employing the use of plant products. Securidaca longepedunculata (SL) serves as a major component of anticancer decoctions in Nigeria. The present study evaluates the cytotoxic potential of the crude extract and fractions of SL root bark against human breast adenocarcinoma (MCF‐7) cell line. The root back of SL was pulverized and extracted with 80% methanol to yield a crude extract which was then submitted to liquid–liquid fractionation with dichloromethane (DCM) and butanol (BUT). The extract and fractions were subjected to Gas Chromatography‐Mass Spectrometry (GC‐MS). MCF‐7 cell line was treated with graded concentrations (9.77 – 5000 μg ml−1) of the aqueous methanol crude extract as well as the DCM and BUT fractions for 24 or 48 hours. Cell viability was thereafter measured by XTT proliferation assay. The activity of the aqueous methanolic crude extract was found to be both dose‐dependent and time independent with IC50 values of 2241 μg ml−1 and 1808 μg ml−1 after 24 and 48 hours respectively. However, the DCM and BUT fractions were significantly (p < 0.05) cytotoxic with IC50 values of 86.27 μg ml−1 and 12.08 μg ml−1 respectively after 48 hours of treatment. The study lends credence that BUT fractions of SL may contain lead agent with anticancer properties and that informed its use in alternative medicine.

Support or Funding Information

SELF SUPPORT
Diet‐derived gallated catechins prevent TGF‐b‐mediated epithelial‐mesenchymal transition, cell migration and vasculogenic mimicry in chemosensitive ES‐2 ovarian cancer cells
Authors: Sicard Audrey-Ann , Annabi Borhane
Published: April 2020

Background

Transforming growth factor (TGF)‐β triggers ovarian cancer metastasis through epithelial‐mesenchymal transition (EMT). Whereas drug design strategies targeting the TGF‐β signaling pathway have been envisioned, the anti‐TGF structure:function aspect of chemopreventive diet‐derived catechins remains unexplored.

Methods

We assessed the effects of eight catechins on TGF‐β‐mediated cell migration using real‐time exCELLigence, induction of EMT biomarkers by RT‐qPCR and immunoblotting, and in vitro vasculogenic mimicry (VM) by 3D‐Matrigel cultures, a process partly regulated by EMT‐related transcription factors.

Results

TGF‐β‐mediated phosphorylation of Smad‐3 and p38 signaling intermediates was more effective in a chemosensitive ES‐2 ovarian cancer cell line but was inoperative in cis‐platinum‐ and adriamycin‐chemoresistant SKOV‐3 ovarian cancer cells. Increases in cell migration and in gene/protein expression of EMT biomarkers Fibronectin, Snail, and Slug were observed in ES‐2 cells. When VM was assessed in ES‐2 cells, 3D capillary‐like structures were formed and increases in EMT biomarkers found. Catechins bearing the galloyl moiety (CG, ECG, GCG, and EGCG) exerted potent inhibition of TGF‐β‐induced cell migration as well as EMT, and inhibited VM, in part through inhibition of Snail and matrix metalloproteinase‐2 secretion.

Conclusions

Our data suggest that diet‐derived catechins exhibit chemopreventive properties that circumvent the TGF‐β‐mediated signaling which contributes to the ovarian cancer metastatic phenotype.

Support or Funding Information

UQAM Foundation and Chaire in Cancer Prevention and Treatment
The effect of tamoxifen on gender differences in unilateral ureteral obstructed rats and human kidney slices
Authors: Tingskov Stine Julie , Jensen Michael Schou , Nørregaard Rikke
Published: April 2020

BACKGROUND

Renal fibrosis is regarded as the most damaging process in chronic kidney disease (CKD) and it is the most consistent predictor of irreversible loss of renal function. To date, there is no good treatment for renal fibrosis. Interestingly, females are less prone to CKD, which might be due to estrogen. We have previously shown that tamoxifen (TAM), a selective estrogen receptor modulator, plays a role for regulation of renal water and salt balance. In this study, we will investigate the effect of tamoxifen on a well‐established in vivo model using unilateral ureteral obstruction (UUO)‐induced fibrosis in female, male and ovariectomized (OVX) female rats. Furthermore, a recently develop model of human renal fibrosis was used to study multicellular pathological processes in human tissue.

METHODS

Fibrosis was induced by 7 days of UUO. Female OVX rats had both their ovaries removed two weeks before UUO. Tamoxifen (25 mg/kg for female rats and 50 mg/kg for male rats; a higher dose of tamoxifen was lethal for female rats) was given by oral gavage starting 5 days before UUO. Tamoxifen treatment was continued for 7 days after UUO. Human precision‐cut kidney slices were exposed to TGF‐β for 48h in order to make the tissue fibrotic and afterwards treated with tamoxifen for 6h. Histological changes were examined by HE staining. Expression of α‐smooth muscle actin, fibronectin, and TGF‐β were evaluated by immunohistochemistry, quantitative PCR, and western blot analysis.

RESULTS

Renal fibrosis was increased after UUO in both male, female and OVX rats. Tamoxifen treatment reduced fibronectin, α‐SMA and TGF‐β gene expression in female UUO rats, but not in male and in OVX rats subjected to UUO. Yet, tamoxifen significantly reduced the protein level of fibrotic markers and the kidney weight in response to UUO in both female as well as OVX and male rats. Furthermore, our data demonstrated that sham operated female and female OVX rats had a higher plasma creatinine level compared to male rats. On the other hand, TAM treated male rats subjected to UUO had a significantly increased urea plasma level, which was not observed in female and female OVX rats. Importantly, treatment with tamoxifen has a tendency to reduce fibronectin in human kidney slices exposed to TGF‐β from women but not in men.

CONCLUSION

These findings indicate that tamoxifen reduces UUO‐induced fibrosis in both female, female OVX and male rats, but have different effects on females and males when it comes to gene levels of fibrotic markers and plasma data. Furthermore, our finding provide preclinical evidence that treatment with tamoxifen affects men and women differently and the concentration of the drug must be adapted to the gender.

Support or Funding Information

This study was kindly supported by Karen Elise Jensen Foundation, the Danish Council for Independent Research, Aarhus University Foundation and the AP Møller Foundation.
Arrestin‐3 interacts with Parkin mutants linked to Parkinson’s disease
Authors: Zheng Chen , Perry Nicole A , Vishnivetskiy Sergey A , Berndt Sandra , Gurevich Eugenia V , Gurevich Vsevolod V
Published: April 2020

Arrestins are multifunctional proteins that were originally identified as suppressors of G protein‐coupled receptors signaling. Later arrestins were found to serve as adapter molecules that bind numerous signaling proteins, such as protein kinases, ubiquitin ligases, deubiquitinases, etc. Here, we showed the direct interaction with arrestin‐3 and parkin, which is a ubiquitin E3 ligase that covalently attaches ubiquitin to specific substrates, including mitochondrial proteins. Parkin is essential for mitophagy – the removal of damaged mitochondria. Many mutations in parkin are linked to familial or sporadic Parkinson’s disease. We identified several binding hot spots on both arrestin‐3 and Parkin using high throughput peptide arrays. Some pathogenic parkin mutations map to these arrestin‐binding hot spots. One of the pathogenic mutations, R275W, significantly increased parkin binding to arrestin‐3 in cells, as compared to wild type parkin. Our data indicate that arrestin‐3 may participate in parkin‐mediated physiological processes that likely contribute to Parkinson’s disease.

Support or Funding Information

This work was supported by grant R35 GM122491 from the National Institutes of Health to VVG.
Loss‐of‐Function Aldolase C Hypermethylation Cause Serotonin Production in Glioblastoma
Authors: Chang Yu-Chan , Hsiao Michael
Published: April 2020

Aldolase involves the glycolysis procedure to maintain glucose homeostasis and further produce some intermediates or energy synthesis. Aldolase C (ALDOC) is specific to the brain region and is activated when the brain develops, is injured or traumatized. Recently, we investigate that ALDOC had independent prognostic role in GBM, which was suppressed in low‐grade glioma and glioblastomas (GBM) and correlated with IDH1 mutant status, MGMT methylation and survival rate. We further found that ALDOC has CpG island in promoter region and was hypermethylated in GBM cells. We collected mRNA level and methylation state of ALDOC by RNA‐sequencing and bisulfite‐specific PCR from Cancer cell line encyclopedia (CCLE) database, respectively. The scatter plots performed the inverse correlation trends between methylation with expression level in GBM cells (n=39, spearman rho=−0.774, P=7.68e‐09). We further dissected the metabolomics pattern and compared with methylation status of ALDOC. The results showed serotonin as one of significant change in GBM cells, the production rate was increased when ALDOC hypermethylation or low expression level. Several malignant GBM cells treated 5‐aza‐2′‐deoxycytidine (5‐aza‐dC) to suppress DNA methyltransferase activity. We found that ALDOC expression levels were restored and migration/invasion ability was reduced. Similarly, the serotonin concentration has attenuated, depending on the ALDOC methylation status. We further computational analyzed the mRNA level of ALDOC correlated with IC50 of temozolomide (TMZ). T98G cells have been selected for ALDOC overexpression and the IC50 of TMZ is reduced by ALDOC induction. In contrast, we transfected shRNAs to knock down ALDOC expression levels in U‐87MG cells with an increase in IC50. The combination of selective serotonin inhibitor and 5‐aza treatment has a synergistic effect on IC50 of TMZ in GBM cells.
The acquisition of positional information in the developing cochlea
Authors: Munnamalai Vidhya , Young Caryl , Thompson Matthew , Fister Alexandra , Umulis David , Fekete Donna
Published: April 2020

The sensory epithelium of the mammalian murine auditory organ, the organ of Corti, is an exquisite example of how specialized asymmetry dictates auditory function. Across the radial axis, the organ consists of three rows of outer hair cells to one row of inner hair cells, which are innervated by efferent and afferent neurons. The mechanosensory inner hair cells that reside in the neural compartment detect sound whereas the outer hair cells in the abneural compartment modulate sound levels. A complex combination of spatio‐temporal signaling inputs across the radial axis endows cells with positional information to take on specific cell fates.

The Wnt and Bmp pathways are two potential signaling mechanisms that influence this radial asymmetry in an opposing manner. We analyzed the requirement of the Bmp pathway for setting up cochlear patterning. Smad4 cKOs showed a loss of the abneural compartment. Prolonged Bmp inhibition decreased the formation of the abneural compartment and outer hair cells. Bmp was required for determining the abneural boundary of the sensory domain. Immediately after hair cell differentiation, Bmp signaling was no longer required. Bmp is necessary, but not sufficient for patterning the entire epithelium. Activation of the Wnt pathway leads to a selective expansion of the neural compartment and inner hair cells. This selective neural expansion was also conserved in the chicken cochlea. Signaling crosstalk of these pathways in a spatiotemporal manner is necessary for proper patterning of the organ of Corti.

Support or Funding Information

R21DC016376R01DC002756R01GM132501
Transcriptional Heterogeneity of Neural Stem Cells during Axolotl Salamander Spinal Cord Regeneration
Authors: Monaghan James , Jensen Tyler , Stein David , Mun Evan , Maden Malcolm
Published: April 2020

Unlike humans, axolotl salamanders are capable of restoring sensory and motor function after spinal cord injury. Upon tail amputation or spinal cord transection, endogenous neural stem cells (NSCs) increase proliferation and differentiate into multiple neural and glial cell types of the central and peripheral nervous system. How NSCs transition from a resting state to a regenerative state and coordinate proper neural differentiation is poorly understood. Here, we study the global changes that occur in NSCs after spinal cord injury as well as specific changes in gene transcription as the NSC regenerative program unfolds. We first studied global changes in NSCs after injury by measuring global RNA synthesis temporally and spatially through incorporation of 5‐ethynyl‐uridine into nascent RNA. Substantial upward shifts in overall transcriptional output occurred in specific NSC populations during regeneration, which was correlated with loss of facultative repressive chromatin marks, and was dependent on Hippo‐YAP signaling. We also found an evolutionarily conserved upregulation of nascent RNA production in active neural progenitors in axolotls and mice. In order to dig deeper into neural stem cell dynamics during regeneration, we adopted multiplexed fluorescence in situ hybridization (FISH) in tissue section and whole mount tissues. We produced a user‐friendly, sharable, Docker Container for automated high‐throughput design of Third Generation Hybridization Chain Reaction (HCR) FISH probe sets. An advantage of HCR‐FISH is that it utilizes non‐enzymatic signal amplification, while maintaining low background signal, leading to high signal‐to‐noise detection of mRNAs. We demonstrate that the HCR‐FISH method is amenable to multiplexing through five sequential rounds of hybridization (SeqFISH). Using our probe design pipeline and HCR‐FISH, we imaged NSC activation and differentiation in tissue sections collected after injury using 15 simultaneously‐detected probe sets indicative to CNS and PNS cell types. Whole mount HCR‐FISH for three neural markers were also imaged simultaneously to assess NSC and neuron responses after injury. Overall, the genetic neuroanatomical mapping performed here using molecular classification of NSCs provides novel information on how NSCs respond to injury to repair a CNS injury.

Support or Funding Information

This project was funded by grants NSF 1656429, NSF 1558017, and NIH OD024909
Integrated Histology‐Pathology Medical Education: What is it Good For?
Authors: Pinder Karen E
Published: April 2020

Medical schools around the world now teach foundational anatomical sciences using integrated methods that have improved the content relevance in medical training and are more consistent with physicians’ clinical reasoning and practice. As stewards of one of these foundational sciences, histology experts at the University of British Columbia (UBC) medical school have embraced a renewed and integrated curriculum. Specifically, we have restructured our undergraduate medical curriculum and implemented an innovative competency‐based integration of years one and two histology with a more perceptibly clinical discipline, pathology. Results from faculty experiences, student outcomes on formative integrated examinations and educational research data from student learning surveys demonstrate the pedagogical effectiveness of our foundational science‐clinical integration.

The next step is to establish how this pre‐clinical integration allows our medical students to utilize foundational science knowledge in subsequent clinical training. That is, how can we demonstrate that the relevance and application of this integration extends beyond the pre‐clinical curriculum and into the clinical years? In other words, what is integrated histology‐pathology medical education good for?

This session will explore one answer to this question by describing our method of mapping histology‐pathology milestones and exit competencies to clinical knowledge and skills in a manner that illustrates the importance of integrated teaching in medical education. That is, by using milestones and exit competency‐based clinical scenarios, we are able to demonstrate how our integrated foundational (histology) and clinical (pathology) medical science approach enables students to progress to the level of clinical expertise expected of all graduating physicians.

Our methods, experiences and outcomes can extend to other foundational science/clinical disciplines partnerships, and this session will prove useful for both educators and clinicians who are exploring or implementing interdisciplinary integrations in both undergraduate and post‐graduate medical training.
Therapeutic Strategies for Full‐Thickness Wounds: Skin and Corneal Tissue‐Engineered Substitutes with Stem Cells
Authors: Germain Lucie
Published: April 2020

The LOEX centre of Laval University is specialized in stem cells and tissue engineering. In addition to fundamental research, LOEX focuses on translational medicine to transfer the discoveries into better treatment for the patients. The LOEX self‐assembly approach of tissue engineering was designed to elaborate complex human living tissue substitutes. Autologous self‐assembled skin substitutes (SASSs) are produced to treat wounds of severely burned patients. The procedure is based on primary culturing dermal fibroblasts and keratinocytes from a few cm2 skin biopsy taken from an unaffected site. Then, dermal fibroblasts are cultured in the presence of ascorbic acid. They secrete and organize their own extracellular matrix (ECM) on which keratinocytes rebuild the epithelium. SASSs exhibit a well‐developed stratified cornified epithelium. The basement membrane is structured and favors long‐term preservation of stem cells in the basal layer of the tissue‐engineered epidermis. The dense collagen network of the tissue‐engineered dermis provides resistance and allow SASS manipulation. After transplantation of autologous SASSs on burn wounds, the graft take was excellent (90–100%). No significant contraction was observed in vivo after grafting. The presence of the ECM promoted a good healing and skin suppleness. SASS presented a well‐organized epidermis and stem cells were settled and maintained in the basal layer after grafting. A complete basement membrane with numerous hemidesmosomes was observed before and after grafting, supporting a strong cohesion between the dermis and epidermis. The integrity of the transplanted SASS persisted over time (1‐ to 8‐year follow‐up) with no defect in epidermal regeneration and no significant contracture. Minimal hypertrophic scars were only observed between SASS. Thus, SASSs provide permanent skin replacement for severe burns. These living substitutes comprising cells and extracellular matrix possess the capability of expanding with the child’s growth. We conclude that the TES produced by this approach is a promising skin substitute for resurfacing full‐thickness skin injury given its functional characteristics: minimal contraction after grafting and promotion of long‐term tissue regeneration. LOEX also used cultured epithelial corneal autografts (CECA), produced from a small biopsy of the limbal area containing stem cells, for the treatment of limbal stem cell deficiency. The first clinical trial is completed, 14 of the 15 patients treated improved their visual conditions. These regenerative medicine technologies pave the way for better treatment of rare diseases.

Support or Funding Information

Supported by the Canadian Institutes for Health Research (CIHR), The Stem Cell Network, Fonds de Recherche du Québec en Santé (FRQS), Fondation des Pompiers du Québec pour les Grands Brûlés and the Cell and Tissue Therapy Network of the FRQS. LG is holder of the Canada Research Chair on Stem Cells and Tissue Engineering of CIHR.
The Anatomy of Cutaneous Wound Healing and Its Inhibition: From Mechanisms to Therapy
Authors: Tomic-Canic Marjana
Published: April 2020

Cutaneous wound healing integrates multiple cellular processes (inflammation, migration, proliferation, barrier formation, matrix deposition and remodeling; differentiation) and harnesses function of many cell types (keratinocytes, fibroblasts, adipocytes, melanocytes), their cross‐talk to endothelial cells, macrophages, mast cells, platelets, peripheral neurons, local and circulating progenitors with a single goal to repair the tissue and restore barrier. Such organized process is tightly controlled in spatial and temporal manner by using complex signaling networks and micro‐environmental factors. Although this ability of skin to maintain its barrier and heal wounds is evolutionary conserved and maintained throughout a lifetime of an organism, it may fails due to various underlining issues. Chronic wounds, such as diabetic foot ulcers (DFUs) and venous leg ulcers (VLUs) represent a widespread unmet clinical challenge and major burden on healthcare systems worldwide. Development of diagnostic tools and treatment strategies is very slow in part due to lack of knowledge regarding the mechanisms of wound healing and its inhibition. We are using genomic approaches and patients’ biopsies to decipher molecular pathophysiology of these chronic wounds as well as molecular mechanisms by which various therapeutic approaches contribute to reprogramming of non‐healing into healing phenotype. Recent findings from primary cells generated from patients as well as transcriptional and mRNA/miRNA profiling revealed multiple new mechanisms that inhibit healing in patients and facilitated identification of wound tissue with repair capability. This chronic wound “genomic signature” includes key mediators of inflammatory response, DNA damage, and response to infection, and specific crosstalk between multiple cell types, revealing new targets and therapeutic approaches.

Support or Funding Information

NIH AR060562, NR015649, NR013881 and DK119085
Protein glycosylation in neural crest epithelial‐to‐mesenchymal transition and migration
Authors: Nie Shuyi
Published: April 2020

Neural crest epithelial‐to‐mesenchymal transition (EMT) and migration involves coordinated remodeling of cell‐cell and cell‐matrix adhesions. During this cell phase transition event, interactions between adhesive molecules need to be dynamically regulated. We recently explored the involvement of protein glycosylation in this process. Protein glycosylation is one of the most complexed, yet least studied types of post‐translational modification. By attaching various forms of oligosaccharide (glycan) on different sites of a protein, glycosylation can regulate protein function by modulating its interaction with other molecules. Through the employment of chemical inhibitors for protein glycosylation, we demonstrated that glycosylation is required for neural crest EMT and migration in frog embryos. To gain a comprehensive understanding of how different proteins are glycosylated during this process, we performed a glycoproteomic analysis at different stages of neural crest development. Through this approach, we identified numerous membrane and secreted proteins that are differentially glycosylated, which may contribute to the dynamic changes in cell‐cell and cell‐matrix interactions during neural crest EMT and migration. We focused on E‐cadherin, a well‐known player in EMT and cell migration, to determine the function of specific protein glycosylation in protein activity and consequently in neural crest cell behavior. Our analysis identified two glycosylation linkage sites on E‐cadherin. The two glycosylation sites were mutated individually or in combination, and the function of mutant E‐cadherin was compared with that of wild type E‐cadherin. Our results showed that mutant E‐cadherins were less effective in promoting neural crest cell migration and this was partly due to an increase in the strength of cell‐cell adhesion mediated by the mutan t proteins. Therefore, our findings suggest a new mechanism to fine‐tune the activity of cell adhesive molecules during EMT and migration.

Support or Funding Information

NIH R00 DE022796, Georgia Tech Startup Fund
Emerging Organ Models and Organ Printing for Regenerative Medicine
Authors: Khademhosseini Ali
Published: April 2020

Engineered materials that integrate advances in polymer chemistry, nanotechnology, and biological sciences have the potential to create powerful medical therapies. Our group aims to engineer tissue regenerative therapeutics using water‐containing polymer networks called hydrogels that can regulate cell behavior. Specifically, we have developed photo‐crosslinkable hybrid hydrogels that combine natural biomolecules with nanoparticles to regulate the chemical, biological, mechanical and electrical properties of gels. These functional scaffolds induce the differentiation of stem cells to desired cell types and direct the formation of vascularized heart or bone tissues. Since tissue function is highly dependent on architecture, we have also used microfabrication methods, such as microfluidics, photolithography, bioprinting, and molding, to regulate the architecture of these materials. We have employed these strategies to generate miniaturized tissues. To create tissue complexity, we have also developed directed assembly techniques to compile small tissue modules into larger constructs. It is anticipated that such approaches will lead to the development of next‐generation regenerative therapeutics and biomedical devices.
The importance of T and B cell immunity in the production of autoantibodies to the angiotensin II type I receptor and the pathophysiology associated with preeclampsia
Authors: LaMarca Babette
Published: April 2020

Preeclampsia (PE), is estimated to affect 5–7% of all pregnancies in the US, is characterized by placental ischemia, chronic inflammation, endothelial dysfunction, systemic vasoconstriction, renal dysfunction, hypertension and IUGR. Despite being the leading cause of maternal and perinatal morbidity and death worldwide, the mechanisms underlying the pathogenesis of hypertension during PE remain unclear. PE women have elevated cytokines, autoantibodies activating the angiotensin II type I receptor (AT1‐AA), cytolytic Natural Killer cells and THelper 1 cells. We have shown that adoptive transfer of CD4+THelper 1 cells increases cytokines, B cells secreting autoantibodies leading to activated cytolytic NK cells and hypertension during pregnancy. Moreover, infusion of purified AT1‐AA activates renal and placental NK cells to cause placental and renal mitochondrial (mt) oxidative stress and dysfunction which contributes to many clinical features of PE including hypertension. Utilizing the Reduced uterine perfusion pressure (RUPP) pregnant rat, a model of PE, we have demonstrated placental ischemia to be an important stimulus for hypertension and many immune mechanisms involved in PE. One important unanswered question in PE research is the importance of B cells in the pathology of the disease, specifically memory B2 cells vs B1 cells in the production of AT1‐AA. Overall our data demonstrate the importance of immunological memory and THelper 1 cells in pathophysiology of PE. In addition to our published data I will present recent findings demonstrating the importance of B cells to secrete AT1‐AA causing hypertension and many clinical features associated with PE.

Support or Funding Information

RO1HD067541 and P20GM121334
Self‐sustaining Laboratories and Collaborative Mentoring in Anatomy Research
Authors: Salinas-Alvarez Yolanda , Quiroga-Garza Alejandro , Elizondo-Omaña Rodrigo Enrique , Guzman-Lopez Santos
Published: April 2020

The development of new technologies and tools for teaching anatomy and the high cost of anatomy laboratories have forced a tendency to decrease the use of bodies and practice. However, these are essential for teaching gross anatomy, and their value is indisputable; students can understand anatomy and develop surgical and clinical skills. Our laboratory has been structured in a manner it can cover all student activities, but also be self‐sustaining through the implementation of postgraduate courses, host for clinical practices for specialists, and pharmaceutical sponsored courses. The incomes are then re‐invested in furthering the laboratory equipment (ultrasound, fluoroscopy, arthroscopy, microsurgery, minimally invasive surgery), to offer a wider range of courses for physicians to expand their knowledge and clinical practice.

These equipments for the laboratory are also used to offer courses for near‐peers that aid in the Anatomy Department. Courses are also made specifically for the anatomy research group (GIA – Grupo de Investigación en Anatomía) students, oriented to areas of interest. Through a tightly organized and collaborative mentoring structure, the group with 17 years of experience, has grown and inspired its members into research. Mentors and mentees benefit from scientific production, enhanced research skill training, and combined experience. Scientific output has increased steadily over time, demonstrating an effective method with longitudinal results of its effectiveness. The group promotes scientific publications in indexed journals, with the students as first authors. The mentor‐mentee relationships are nurtured causing appreciation and recognition bilaterally. Most students wish to continue with a clinical residency, however, most report they wish to continue with some form of clinical research.

Support or Funding Information

None
Impact of the GH‐IGF Axis on Adipose Tissue and Obesity: Are there Benefits of Endocrine Defects
Authors: Kopchick John
Published: April 2020

Disruption of the mouse growth hormone receptor gene (GHR−/−) results in dwarf, obese, GH resistant mice that possess low levels of IGF‐1 and high levels of GH with intense insulin sensitivity. These mice are similar to humans with Laron Syndrome (LS) in which the individuals are small in stature with low IGF‐1 and high GH levels. Cancer and diabetes rates in both the GHR−/− mice and LS individuals are low or non‐existent. In this regard, GHR−/− mice have extended longevity, in fact, they are the longest‐lived laboratory mouse. Thus, a defect in the action of GH may be beneficial for increased health‐span and longevity. In attempts to dissect which, if any, of the insulin sensitive tissue contribute to the extended health and lifespan of these mice, we have individually disrupted the GHR gene in liver, muscle, and adipose tissue. Surprisingly, only muscle specific GHR gene disruption resulted in a modest increase in life span while none of the others have the impressive extended longevity seen in GHR−/− mice. Since a decrease in the GH/IGF‐1 axis is important for extended health and lifespan, the ability to down regulate its action in adult life may be beneficial. To that end we have, and are, attempting to disrupt the mouse GHR in adult life. To date, disruption at 6 weeks of age results in female mice with extended longevity. Experiments to ‘knock‐out’ the GHR gene at 6 months and one year are ongoing. Also, since adipose tissue (AT) is metabolically important, data regarding the location and function of AT in these mice will be presented.

Support or Funding Information

This work was and supported by the State of Ohio’s Eminent Scholar Program that includes a gift by Milton and Lawrence Goll, the AMVETS, and NIH R01AG059779.
Neuroanatomical characterization of angiotensin‐sensitive neurons within the paraventricular nucleus of the hypothalamus reveals an access point for controlling behavioral and physiological responses to stress.
Authors: Krause Eric G.
Published: April 2020

Stressors perceived in the environment or those arising from the internal milieu create neural signals that converge on the paraventricular nucleus of the hypothalamus (PVN). The PVN contains distinct neuronal phenotypes that integrate and transduce these signals into cardiovascular, neuroendocrine and behavioral responses that cope with the impact of psychological and physiological stress. Accumulating evidence implicates angiotensin receptors within the PVN as mediators of the stress response; however, the neuroanatomical mechanisms underlying these effects are not completely understood. Here, we use the Cre‐LoxP system in mice with virally‐mediated gene transfer and optogenetics to characterize the structure and function of neurons in the PVN that express the angiotensin type 1a receptor (Agtr1a). Neuroanatomical experiments discovered that Agtr1a(s) were expressed on neurons in the PVN that synthesize mRNAs for corticotropin‐releasing‐hormone (CRH) or thyroid‐releasing‐hormone (TRH). These CRH and TRH neurons were found to send direct axonal projections to the median eminence and their optogenetic excitation increased, but inhibition decreased, circulating levels of corticosterone and thyroxine, indicating that Agtr1a‐expressing neurons in the PVN control the activity of neuroendocrine axes. To investigate how angiotensin signaling may affect stress responding, we engineered mice with Agtr1a specifically deleted from the PVN (Agtr1a‐PVN KO). Under basal conditions, no effect of the deletion was observed; however, maintenance on high fat diet, a metabolic stressor, revealed a gene x environment interaction whereby Agtr1a‐PVN KO mice had increased adiposity and body weight but lower blood pressure relative to controls. In psychogenic stress paradigms, Agtr1a‐PVN KO mice were found to have slight but significant decreases in anxiety‐like behavior and cardiovascular reactivity. Collectively, these results suggest that stimulation of Agtr1a(s) in the PVN contribute to activation of neuroendocrine axes that mediate behavioral and physiological responses to metabolic and psychogenic stressors.

Support or Funding Information

NIH: HL122494, HL139868, HL136595
Hypothalamic Regulation of Blood Pressure
Authors: Lazartigues Eric D.
Published: April 2020

In addition to brainstem nuclei well‐known to be involved in baroreflex regulation of blood pressure, several hypothalamic regions have also been shown to modulate autonomic regulation and cardiovascular function. Among them, the paraventricular nucleus (PVN) comprised of magnocellular neurons involved in vasopressin and oxytocin secretion and parvocellular neurons involved in pre‐sympathetic activity. Elevated firing activity of pre‐sympathetic PVN neurons is thought to contribute to hypertension. The PVN also contains components of the renin‐angiotensin system (RAS), like the angiotensin‐II type 1 receptor (AT1R) which expression is enhanced in hypertensive conditions. We previously reported that Angiotensin converting enzyme 2 (ACE2) is also expressed in the PVN and capable of metabolizing Ang‐II levels thus reducing Ang‐II pro‐hypertensive signaling. More recently, we identified ADAM17 as a critical mediator of AT1R downstream signaling, supporting the maintenance of hypertension and neuro‐inflammation. We observed that during RAS overactivity, AT1R stimulation leads to translocation of ADAM17 from the cytoplasm to the plasma membrane, leading to enhanced activity of this sheddase. As a result, ACE2 is cleaved by ADAM17 from the plasma membrane and the compensatory activity of the enzyme is thus compromised, promoting an imbalance of autonomic function and favoring the development of neurogenic hypertension. Recently, we investigated the interaction of AT1R, ADAM17 and ACE2 on PVN pre‐sympathetic neurons and hypothesized that ADAM17 would impair ACE2 regulation of neuronal activity on these cells, leading to an increase in sympathetic outflow and eventually blood pressure. We also attempted to identify patterns of expression for these RAS components on specific PVN neurons. To address these questions, we relied on novel transgenic mouse models with selective deletion of RAS components in the PVN and electrophysiology. Our observations suggest that ACE2 maintains inhibitory neurotransmission into the PVN and its expression is prevalent on inhibitory neurons. On the other hand, ADAM17 is critical for Ang II‐mediated activation of kidney‐related presympathetic glutamatergic neurons in the PVN, confirming the supportive role of ADAM17 in increasing presympathetic neuronal activity. Balancing inhibitory and excitatory neurotransmission from the PVN is essential to prevent a sympathetic overdrive and our observations provide new targets to modulate excessive sympathetic activity in hypertension.

Support or Funding Information

NHLBI HL093178.
A Sequenced and Team Approach to Surface Anatomy Education in Physical Therapy in Preparation for Clinical Practice and Collaborative Care
Authors: Topp Kimberly
Published: April 2020

A deep understanding of anatomy is foundational in the clinical practice of physical therapy, and is key to discovery and scholarly work in movement sciences. Similar to several other health science professions, education of the physical therapist begins with prerequisite coursework in an undergraduate program, continues with doctoral‐level courses in a graduate program, and often post‐licensure residency or fellowship training. Anatomy and physiology are prerequisites for many DPT programs, and anatomy is included in the required elements for accreditation of DPT programs, residencies and fellowships. Surface anatomy is foundational for clinical work in physical therapy and is honed over years of practice.

In the DPT curriculum, surface anatomy is incorporated into early courses in gross anatomy and skills acquisition, including training in goniometry and assessment of muscle strength and vital signs. Surface anatomy continues into advanced gross anatomy courses and education in physical examination, neuromusculoskeletal assessment, and movement science. Early on, students identify bony landmarks, arterial pulses and muscle bulk, similar to early learners in medical curricula. As they progress, DPT students learn to observe subtle contours and shadows, identify muscle weaknesses and joint restrictions through observation of movement, assess muscle contraction and tendon motion using ultrasound imaging, and palpate delicate superficial nerves, millimeter differences in joint motion, and directional restrictions in layers of fascia.

Licensed physical therapists gain additional knowledge of anatomy through continuing education courses, residency and fellowship training. Courses may include advanced training in such areas as pelvic health, oncological care, prosthetics, or vestibular rehabilitation. Residencies and fellowships enable clinical specialization, such as in pediatrics, orthopedics or cardiopulmonary care. Each of these courses and specialty certifications includes advanced training in surface observation of underlying anatomy.

In this talk, Dr. Topp will describe a sequenced approach to surface anatomy education in physical therapy, and how surface anatomy is used in clinical physical therapy. As budding experts in anatomy, students in physical therapy may provide guidance in surface anatomy for physicians in training, and the talk will include examples of near‐peer learning of surface anatomy and case‐based musculoskeletal examinations.

Support or Funding Information

No funding was required.
‘It All Begins with the Outside of the Body, with a Person Wrapped in Their Own Skin’: How to Make Surface Anatomy a Central Part of a Curriculum
Authors: Tunstall Richard
Published: April 2020

The majority of patient examinations and interventions within a healthcare setting rely on the use of surface anatomy; why, therefore, do so many of the courses used to train healthcare practitioners place little emphasis on ensuring proficiency, or indeed mastery, in the surface landmarking and localisation relevant to safe practice? Despite the regularity with which it is used in practice, surface anatomy is often treated as a second‐class relation to gross anatomy, as an add‐on component that students will figure out for themselves via logical deduction. To further add to the issue, many current texts present outdated descriptions of surface anatomy and fail to recognise that there is a range of normal positions within which structures may sit.

Arguably, surface anatomy should form one of the core components of any clinical anatomy and medical imaging curriculum for healthcare practitioners and all graduating practitioners should be able to consistently locate and visualise structures relevant to their practice via the skin. This presentation will discuss a range of techniques, methods and resources that have be used to enable surface anatomy to become an explicit and central part of undergraduate and postgraduate training, and will highlight the challenges and outcomes of implementing such an approach.
Use of Modeling Clay as an Inexpensive Educational Tool to Improve Student Understanding of the Anatomy of the Human Perineum
Authors: Castilla Marina , Whitney Jennifer
Published: April 2020

The human perineum is a complicated area of anatomy for graduate and medical students to master. Dissection/prosection of this area is time consuming and often yields minimal results. For graduate students taking a non‐dissection gross anatomy course, we have used an alternative three‐dimensional modeling method which may facilitate student learning and potentially serve as an adjuvant or replacement for cadaveric dissection of this region. To assess the activity we measured multiple‐choice question performance and student self‐reported confidence of subject matter knowledge. Student performance on multiple‐choice questions before and after the activity was assessed and showed an over 20% improvement. Participants in the modeling activity also scored on par with medical students that perform cadaveric perineum dissection. Overall student confidence in their knowledge improved slightly. Building a perineum out of modeling clay may be a useful tool for non‐dissection anatomy courses and may be further investigated as a potential replacement to cadaveric dissection of the perineum.