bluuebunny/crossref_metadata_2025_split huggingface dataset:
Native American and First Nation Perspectives on the Use of Human Remains in Education & Proximity to Death
Authors: Reynolds Amberly M. , Pennefather‐O’Brien Elizabeth 
Published: May 2022

Cultural attitudes regarding interactions with the deceased, the dead and contact with the lifeless human body vary throughout the world, including among Native American, First Nation, Metίs, and Inuit groups in North America. From remains that have been obtained without consent to working with cadaveric specimens, this talk addresses the beliefs and concerns of indigenous peoples regarding the use of human remains as educational specimens and interaction with the dead. Both speaker perspectives as well as preliminary results from research will address the diverse attitudes towards the dead, treatment of the dead, use of human remains for teaching and research purposes. We will also acknowledge the principle of informed consent in a colonial world. Anatomical education is often reliant on the gracious donation of humans but what are we to do if they were not obtained with consent as are many of the indigenous specimens still maintained at universities?Working with human remains can also be of concern for indigenous students with respect to their cultural beliefs. This talk will address the importance and considerations needed for indigenous students while providing the audience with a basic understanding of some of those cultural beliefs. It is not only important to have respect for the human remains but also for the students who encounter them. Anatomical educators must have an awareness of cultural and religious beliefs that are at odds with working with human remains or being in the presence of a lifeless body, so that we are prepared to provide inclusive learning environments for our students. We hope to provide an inclusive discussion on Native American and First Nation perspectives on the use of human remains in education and the proximity to death.
A novel NSC small molecule inhibitor inhibits proliferation of triple‐negative breast cancer cells through metabolic reprograming
Authors: Yousefi Hassan , Lauterboeck Lothar , Okpechi Samuel , Zabaleta Jovanny , Guidry Jessie , Yang Qinglin , Alahari Suresh 
Published: May 2022

BackgroundTreatment of patients with triple‐negative breast cancer (TNBC) has been challenging due to the absence of well‐defined molecular targets and high invasive and proliferative capacities of these cells. Current treatments against TNBC have shown minimal activity due to the high recurrence rate in the patients. There is, therefore, a pressing need for novel and efficacious therapies against TNBC. Here, we found a novel small molecule inhibitor with potent anti‐tumor activity against TNBC cells.MethodsAnti‐proliferative effects of NSC small molecule inhibitor were determined using 2D and 3D culture cell proliferation assays. Using proteomics, NGS and enrichment analysis, we globally investigated top regulatory pathways affected by this compound in TNBC cells. Subsequently, we validated the proteomics and NGS analysis data using seahorse and enzymatic assays. Finally, we showed the inhibitor anti‐tumor effects in vivo. Data were evaluated in triplicate against untreated control cells. Data were graphed and analyzed using GraphPad Prism Software 8.0 using one‐way ANOVA and the unpaired two‐tailed Student’s t‐test.ResultsIn this report, we showed that a novel NSC small molecule inhibitor reduced the proliferation of TNBC cells. Proteomic analysis confirmed a significant metabolic reprograming including suppression of glycolysis and oxidative phosphorylation after treatment. Furthermore, we found that treatment with NSC small molecule inhibitor impaired glycolysis and oxidative phosphorylation via modulating the activity of metabolic regulator enzymes.ConclusionsAltogether, our data indicate that NSC small molecule inhibitor may exhibit anti‐tumor activity in TNBC cells and provide a rationale for further investigation of the potential of NSC small molecule inhibitor as an attractive therapeutic drug for TNBC. Further in‐depth investigations are required to evaluate the exact targets and mechanism of this potent small molecule inhibitor, and its anti‐neoplasm effects on TNBC in vivo.
Mammalian testes utilize multiple modalities to moderate spermatogonial responsiveness to retinoic acid
Authors: Johnson Taylor A. , Kirsanov Oleksandr , Niedenberger Bryan A. , Harrington Ellen , Malachowski Taylor , Geyer Christopher B. 
Published: May 2022

What is knownThe foundation of mammalian spermatogenesis is provided by undifferentiated spermatogonia that proliferate before responding to retinoic acid (RA). RA converts undifferentiated into differentiating spermatogonia committed to entering meiosis. This RA‐regulated fate transition is essential for maintaining normal ratios of spermatogonia during steady‐state spermatogenesis in adult testes. Imbalances in spermatogonial populations often result in infertility – enhanced differentiation leads to eventual germline depletion, while impaired differentiation increases the likelihood of developing testicular cancer.Study ObjectiveDefine the extent cell autonomous and non‐autonomous mechanisms in adult mammalian spermatogonia regulate their response to RA.HypothesisOnly spermatogonia primed to differentiate will respond to RA and precociously progress through spermatogenesis.MethodologyAdult mice were treated in vivo with vehicle (DMSO), RA, or talarozole (RA catabolism inhibitor) and euthanized 12‐48 hours later. Testes were removed and various assays used to assess populations for responsiveness to RA and capacity to precociously differentiate and enter meiosis. Concurrently, a model of synchronized spermatogenesis was used to define the potential of sequential subsets of differentiating spermatogonia to respond to RA and precociously enter meiosis. In complementary ex vivo experiments, mouse testes and patient‐derived testicular biopsies were cultured with DMSO‐alone, RA, and/or talarozole for 6‐24 hours to evaluate RA responsiveness and changes in spermatogonia fate using immunostaining and qRT‐PCR.Main resultsExogenous RA elicited a rapid (12 hour) and significant increase in expression of the RA‐responsive gene ‘stimulated by retinoic acid 8’ (STRA8) in undifferentiated (ZBTB16+) spermatogonia. STRA8 expression decreased over the next 36 hours, such that numbers of STRA8+/ZBTB16+ spermatogonia were comparable in testes from vehicle‐ and RA‐treated mice. This transient increase in STRA8 affected spermatogonia at all stages of spermatogenesis. Exogenous RA also elicited STRA8 expression in differentiating (KIT+) spermatogonia in adults and synchronized mice. Despite this capacity for responding to exogenous RA, obvious precocious progression was not observed within the testis. Talarozole treatment elicited STRA8 expression in spermatogonia akin to RA treatments. Finally, qRT‐PCR showed adult mouse and human testes regulate responsiveness to RA similarly, based on mRNA expression profiles.ConclusionIn response to exogenous RA, undifferentiated spermatogonia transiently upregulate expression of RA‐responsive genes, but this did not result in precocious progression into meiosis.ImplicationsThe adult mammalian testis is a well‐buffered system in which exogenous RA has little effect on cell fate, thus preserving the normal timing and progression of spermatogenesis.
Agomelatine Ameliorates Obesity‐Induced Kidney Injury through the Inhibition of Renal Fibrosis and Improvement of Impaired Autophagy
Authors: Promsan Sasivimon , Phengpol Nichakorn , Sutthasupha Prempree , Lungkaphin Anusorn 
Published: May 2022

IntroductionChronic kidney disease is the crucial complication that could occur in obesity. Prolong high‐fat diet consumption can stimulate reactive oxygen species (ROS) and contribute to kidney injury through various pathways. Autophagy plays an important role for maintaining intracellular homeostasis. ROS could induce impaired autophagy, subsequently led to kidney damage. Agomelatine (AGOM), a melatonin analogue, is indicated for the treatment of major depressive disorders. The antioxidant and anti‐inflammation effects have been reported in AGOM treatment in chronic mild stress‐induced depression. Previous study found that AGOM inhibited apoptosis and ER stress in the kidney under high‐fat diet condition in rats. However, the effect of AGOM supplementation on renal fibrosis and impaired autophagy in obese condition has never been elucidated.AimsTo evaluate the renoprotective effect of AGOM on kidney function in high‐fat diet‐induced kidney injury, as well as to explore the underlying mechanisms involving autophagy signaling pathway.Materials and methodsMale Wistar rats were received normal diet (ND) or high‐fat diet (HF) for 16 weeks. Then, the HF rats were separated into 4 subgroups including (1) HF; (2) agomelatine20 (AGOM20); (3) AGOM40, the rats were received AGOM at the dose of 20 and 40 mg/kg/day, respectively; and (4) N‐acetylcysteine (NAC), the rats were received NAC at the dose of 100 mg/kg/day by oral gavage for 4 weeks. After 4 weeks of treatment, all rats were sacrificed. Blood, urine and kidney tissues were collected for further investigations.ResultsThe results demonstrated that HF rats developed kidney dysfunction as shown by the raising in serum creatinine, creatinine clearance along with the changes of glomerular structure and the elevating of positive area of α‐SMA, which is renal fibrosis indicator. These alterations were reversed by AGOM and NAC treatment. Moreover, HF rats showed renal oxidative stress as indicated by the increasing in PKCα and NOX4 expression and decreasing in antioxidant enzyme, GCLC. AGOM and NAC administrations reduced oxidative stress and restored antioxidant enzyme. Furthermore, high‐fat diet consumption markedly reduced AMPK, along with the elevation of mTOR expression. Additionally, we found that HF rats aggravated impaired autophagy which could induce oxidative stress, fibrosis and finally led to kidney cell death. AGOM administration restored autophagy process as shown by the increasing in Beclin‐1, LC3B and Atg5 protein. These results demonstrated that AGOM improved kidney injury through the regulation of oxidative stress, fibrosis and autophagy process.ConclusionsThese findings indicated that AGOM prevented kidney injury under obese condition via the inhibition of oxidative stress and fibrosis and improved impaired autophagy via regulating AMPK‐mTOR‐autophagy signaling pathways.
Regulation of Sphingomyelin Synthesis by Cholesterol
Authors: Kim Yeongho , Burd Christopher G. 
Published: May 2022

The most abundant sphingolipid of the plasma membrane (PM), sphingomyelin (SM), binds to cholesterol in a manner dependent on the structure of fatty acids in the SM, and it is a major determinant of the cholesterol content of the PM. Synthesis of SM in the Golgi apparatus produces an mol‐equivalent of diacylglycerol (DAG), a signaling lipid implicated in the fission of secretory vesicles from the Golgi. We speculate that coupled synthesis of SM and DAG in the terminal compartments of the Golgi results in fission of secretory vesicles enriched in SM and integral membrane proteins that favor sphingolipid and cholesterol‐rich membrane. Two classes of SMs are synthesized in the Golgi apparatus that are distinguished by the length of the fatty acyl chain on the ceramide backbone: long chain (LC; ≤C20), and very long chain (VLC; >C20) SMs. Whereas the precursor to LC‐SMs is supplied to the Golgi via non‐vesicular transport of LC‐ceramide from the ER to the Golgi by ceramide transport protein (CERT), it is speculated that VLC‐ceramide is supplied to the Golgi via bulk membrane trafficking pathways. From the Golgi, LC‐SM and VLC‐SM are trafficked to the PM in SM‐rich secretory vesicles that also contain a select cohort of secreted proteins (Deng et al., 2018).Cholesterol extraction increases sphingomyelin synthesis (Perry and Ridgway, 2006), however, the two classes of SMs are differentially regulated; we discovered that only synthesis of very long chain (>C20) SMs was increased. Furthermore, cholesterol loading reduced the synthesis of VLC‐SM, only. We then asked, what cellular components regulate VLC‐SM synthesis and how do they influence the cholesterol content of the PM and other organelles. Synthesis of VLC‐SM was inhibited by interfering with COPII‐mediated export from the ER and by depletion of ER‐localized ceramide synthase 2, which produces VLC‐ceramide. Reduced VLC‐SM synthesis resulted in the depletion of cholesterol from the PM inner leaflet and a concomitant accumulation in the cytoplasmic leaflets of the membranes of lysosomes and other intracellular organelles. Increased synthesis of VLC‐SMs by cholesterol depletion resulted in increased the level of diacylglycerol (DAG) in the Golgi and the appearance in cytoplasmic vesicles containing TGN46 which is secreted in SM‐rich secretory vesicles. The results suggest that VLC‐SM synthesis is regulated by cholesterol content on the PM and cholesterol depletion‐induced synthesis of VLC‐SM enhances SM‐specific secretory pathway from the Golgi to the PM.
Proteasomes in the Cells of the Ocular Surface in Health and Disease
Authors: Bardag Gorce Fawzia , Wolosin Mario J. , Oliva Joan , Niihara Yutaka 
Published: May 2022

PurposeLimbal stem cell deficiency (LSCD) is associated with corneal conjunctivalization that compromises corneal transparency and causes visual impairment. Previously, we found marked changes in proteasome population in corneas of rabbit with LSCD, and showed immunoproteasome (IPR) presence in central corneas during conjunctivalization compared to healthy rabbit cornea. In the present study, we further map the distribution of proteasomes population in the ocular surface.Methodsgene expression intensity obtained by microarray analysis of human healthy corneas and conjunctiva previously reported by the group of Dr. Wolosin (Turner H.C. et al 2007) was used to conduct this study. Constitutive proteasome (CPR) subunits (B5, B2, and B1) were compared to the inducible immunoproteasome (IPR) subunits (B5i, B2i, and B1i). The ratio between CPR and IPR subunits was calculated for cornea and for conjunctiva.ResultsThe comparison of the ratio B5/B5i, B2/B2i and B1/B1i between cornea and conjunctiva showed a significant high expression of CPR in corneas and a high expression of IPR in conjunctivae. In addition, PA28, the regulatory complex of IPR, and the major histocompatibility complex, class I A and B, were found significantly expressed in conjunctiva. Keratins K3/K12 were specifically expressed in cornea and keratins K4/K13 were specifically expressed in conjunctiva, which could be used to normalize the data analysis.ConclusionThe results indicated that conjunctivae contain high levels of IPR and that during conjunctivalization, corneas lose CPR to express IPR. The ration CPR/IPR could be used as an indication of conjunctivalization level in case of corneal injuries.
Needs Assessment of Essential Anatomy ‐ A Perspective from Primary Care Residents
Authors: Harmon Derek J. , Hankin Mark H. , Martindale James , Niculescu Iuliana 
Published: May 2022

Introduction. Curricular development begins with identifying a problem, followed by a need assessment of the curricular components. Changes in pedagogical approaches and reductions in curricular time for gross anatomy have been identified as a problem. The objective of this study was to conduct a needs assessment of the most essential anatomy from the perspective of primary care residents.Methods. Primary care residents in Family Medicine (FamMed) and General Internal Medicine (IntMed) at Oakland University William Beaumont School of Medicine were recruited for the study. A survey assessed the clinical importance of 907 specific anatomical structures from all anatomical regions on a 1‐4 Likert‐scale (1= “not important and 4 = “essential”). The structures were selected from the bold terms in Gray’s Anatomy for Students, Clinically Oriented Anatomy, and Grant’s Dissector. In the analyses, all 3 and 4 ratings for each structure were summed and divided by the total number of respondents. A structure rated as a 3 or 4 by ≥75.0% of respondents was classified as “essential”, 50.0‐74.9% “more important”, 25.0‐49.9% “less important”, and 0.0‐24.9% “not important”. Kruskal‐Wallis analyses were conducted to assess differences between the two residency groups for each region.Results. Fifty adult primary care residents in FamMed and IntMed completed the survey. In total, 16.7% of structures were classified as essential, 58.9% more important, 24.3% less important, and 0.7% not important. FamMed rated structures categorized as skeleton, nerves, fasciae, anatomical spaces, blood vessels, lymphatics and surface anatomy (p<0.0001), and muscles (p=0.002) significantly higher than IntMed, but there was no significant difference in organs (p=0.012). It was notable that 100.0% of cranial nerves were classified as essential, and 94.5% of surface anatomy structures were classified as essential or more important. There were no statistically significant differences between the two groups of residents for any of the anatomical regions.Conclusion. Conducting a needs assessment is a crucial early step in curricular development. This is the first study to conduct a needs assessment of anatomy curricular content solely from the perspective of primary care clinicians. Surface anatomy was an area with nearly all structures rated as essential or more important, which reinforces the importance of including surface anatomy in pre‐clinical gross anatomy education.Significance. The data from this study provides insight into how clinicians rated specific anatomical structures. Due to the broad variation in medical curricula, the authors suggest that the classifications of importance by these clinicians can serve as a curricular roadmap by anatomy educators for the evaluation, revision, or development of curricula with regard to gross anatomy education in the context of reduced contact hours, new pedagogies, and vertical integration.
Structural Landscape of SARS‐CoV‐2 entry & activation of spike glycoprotein & potential interventions
Authors: Vankadari Naveen 
Published: May 2022

The pandemic COVID19 illness caused by SARS‐CoV‐2, which produces pneumonia and lower respiratory tract infections, is a serious public health concern, with frightening mutations causing over 4.5 million deaths globally. Whilst effective immunisation shows promise globally, several antiviral treatments are being clinically evaluated to fill the “therapeutic gap” in treating infected people. Understanding the entire repertoire of diverse host factors engaged by SARS‐CoV‐2 for entry and pathogenicity is required for long‐lasting potential therapeutics or vaccines. Here, using a structural and molecular approach, we show multistage processing of SARS‐CoV‐2 spike‐protein for virion activation, infection, and how mutations influence it. We solved the structures of spike‐protein in complex with different host cell factors (TMPRSS2, Furin, CD26 (DPP4), and NRP1) with functional activity, and these insights into uncovering how viral spike‐protein engages and primed with these multiple host factors, in addition to ACE2, to hijack host cell entry. Furthermore, our COVID19 patient genome sequencing reveals that allele in TMPRSS2 (V160M) and Furin provided protection from COVID19 infection, and its structural mechanism is further addressed and potential drug clinical trials. Additionally, our large‐scale retrospective cohort studies proved Arbidol and derivatives as potential therapies for COVID19, using structural studies, we demonstrated the mechanism of action of Arbidol in disrupting spike function. These findings cognize the complete mechanism of viral spike‐glycoprotein processing/priming that leads to cascading entry into the host cell, paving the door for future vaccine development and identifying key targets. Our comprehensive, multifaceted research reveals the complexity of the SARS‐CoV‐2 spike‐protein and clinical studies aid in therapies.References: 1. Li, F., et al., Nature Communications 12, 866 (2021). 2. Vankadari, N. & Jacqline, A., Emeging Microbes & Infections, 9(1), 601‐604 (2020). 3. Vankadari, N. Int. Journal of Antimirobial Agents, 56(2),105998 (2020) 4. Andersen, K.G., et al., Nature Medicine 26, 450–452 (2020). 5. Qibin Liu et al., MedRxiv CSH Press (2021), March 22 (2) 6. Ravikanth, V et al., mGENE (2021), 5, e100930. NEJM (Reviewed) 7. Vankadari, N. ACS Journal of Physical Chemistry Letters, 11(16), 6655‐663 (2020).
A Bifidogenic Prebiotic Supplement Beneficially Alters Gut Microbial Composition, Activity, and Plasma Metabolites in Healthy Individuals Consuming Low Fiber Diets
Authors: Kang Jea Woo , Tang Xinyu , Zivkovic Angela M. 
Published: May 2022

A randomized order, double‐blinded, placebo‐controlled cross‐over intervention study was conducted to test the effectiveness of a prebiotic supplement in healthy participants consuming a diet low in fiber. This clinical trial was registered at clinicaltrials.gov as NCT03785860.Fecal microbial DNA was extracted and sequenced with shallow shotgun sequencing for metagenomics. Plasma metabolites were analyzed with untargeted liquid chromatography–mass spectrometry for metabolomics. Statistical analyses were performed with the package edgeR in R version 4.0.2. For each outcome, linear mixed models were used with fixed effects of treatment, treatment order, and their interaction and random effect of individual participants.From the metagenomic analysis, the relative abundance of the genus Bifidobacterium and several bifidobacterial species increased after prebiotic supplementation compared to the placebo. Additionally, gene counts of genes sacA, xfp, xpk associated with prebiotic utilization significantly increased after the prebiotic supplement. Other genes that increased in counts were poxB and ackA, which are related to acetate metabolism. Changes in bifidobacteria abundance were positively correlated with changes in genes related to fructose and choline metabolism, and negatively correlated with a gene that encodes for acetate kinase.Plasma concentrations of indolepropionate significantly increased whereas choline and several acylcholines such as stearoylcholine, dihomo‐linolenoyl‐choline, linoleoylcholine, arachidonoylcholine, palmitoylcholine, and oleoylcholine significantly decreased after the prebiotic compared to the placebo arm.The results of this study demonstrate that a convenient, easy to use bifidogenic prebiotic supplement beneficially altered gut microbial composition and activity, which was reflected in changes in plasma metabolites in healthy individuals consuming low fiber diets.
3’ Terminal end‐labeling of structured RNA by Klenow DNA polymerase
Authors: Mwangi Mary N. , Baird Nathan N. 
Published: May 2022

RNA labeling is a key step in studying RNA localization, folding, and structural dynamics using fluorescence based assays. Current methods of fluorophore attachment to long (>100 nucleotides) and structured RNAs are hampered by low labeling yields and limitations in attachment position. These challenges have led to an interest in developing alternative methods that would enable the site‐specific labeling of long and structured RNAs. To this end, we have developed a site‐specific, chemo‐enzymatic method of fluorophore incorporation to RNAs. This novel method uses Klenow enzyme, a DNA polymerase, to add modified nucleotides to RNAs 3’ end in a templated manner. To facilitate this modified nucleotide addition in structured RNAs, we designed helper DNA oligonucleotides to disrupt any pre‐existing RNA intramolecular structure. Using this method, we have achieved efficient 3’ terminal end‐labeling of chemically synthesized and in‐vitro transcribed RNAs of different lengths with cyanine dyes.This method will be routinely used in fluorescence‐based assays that investigate the structure‐function relationships of RNAs involved in cancer and HIV progression.