bluuebunny/crossref_metadata_2025_split huggingface dataset:
LUDIC ANATOMY PROMOTES LONG‐THERM KNOWLEDGE RETENTION
Authors: Maia Jad Gabriele Silva , Kubrusly Bruna Sobreira , Filho Cezar Nilton Rabelo Lemos , Freitas Francisco Orlando Santos Rafael , Barreto João Erivan Facanha , Cerqueira Gilberto Santos Facanha 
Published: April 2020

The project Ludic Anatomy is a learning and teaching tool used in the Medical, Pharmacology, Dentistry and Biology courses of the Federal University of Ceará. Concerning to the benefits of ludic activities, they make the teaching and learning process more dynamic and interesting, by stimulating the active studying of the muscular system and creating a playful method which is greatly different from the orthodox classes, they also have a positive influence in the memorizing development of anatomical terms. Considering the method of the ludic activity, the students used pins to mark and, then, identify fifty different muscles of the doll. Therefore, the objective of this study is to investigate if the use of toys in the anatomy’s learning and teaching process promotes long therm (6 months) knowledge retention. It was realized a descriptive exploratory study with quantitative approach with 61 medical students who participated of the project. To collect data, a semi‐structured questionnaire was applied at the time of activity and lasted six months. For statistical analysis, the Mann Whitney test was considered significant when presented the p<0,05. The mean age of the participants was 19,97 ± 0,46 for males and 19,74 ± 0,37 for females, there was not an important difference between the ages. A statistically significant difference was observed in the knowledge retention when compared the correct (7,69 ± 0,18) and wrong (1,30 ± 0,18) answers. A negative correlation (R −0,057) between the time spent studying and the numbers of questions right was observed. In conclusion, it was verified that the Ludic Anatomy promotes learning and the long‐therm knowledge retention. That project’s advantage demonstrates the importance of the ludic activity as an effective learning tool in the improvement of the traditional teaching of Anatomy.Support or Funding InformationCNPQ
Investigating the Binding of Albumin and C‐peptide to Red Blood Cells
Authors: Geiger Morgan , Zinn Kurt , Spence Dana 
Published: April 2020

The pancreatic β‐cells of healthy individuals secrete insulin and C‐peptide, a 31‐amino acid peptide, in a 1:1 mole ratio. However, the β‐cells of people with type 1 diabetes (T1D) are damaged or destroyed. Therefore, exogenous insulin is required for people with T1D to survive. Even with regular insulin treatments, patients develop complications such as retinopathy, neuropathy, and nephropathy. Research from our group demonstrates that C‐peptide increases microvascular blood flow, therefore, it could be a useful tool to decrease these complications due to poor blood flow. However, the unknown C‐peptide receptor and mechanism has been a major roadblock in utilizing C‐peptide as a therapeutic. Utilizing ELISA, our group has demonstrated that approximately 1,800 C‐peptide molecules bind per red blood cell (RBC) in the presence of albumin. Without albumin, there was no detectable C‐peptide binding per RBC. Rather than C‐peptide solely binding to a RBC receptor, we hypothesize that C‐peptide binds to RBCs through an albumin/C‐peptide complex receptor. Bovine serum albumin (BSA) binding per RBC was detectable through an attached gamma decay radiolabel (99mTc). A binding saturation experiment was conducted to examine the specific binding of BSA to RBCs with and without C‐peptide. The specific binding curves revealed that albumin saturates at 14,021 ± 1489 BSA molecules/RBC with a Kd of 1.07 (±0.19) × 10−7 M and a Bmax of 1.94 (±0.06) × 10−8 M, or 13,900 receptor molecules/RBC. Whereas, in the presence of C‐peptide and zinc, albumin saturates at 16,696 ± 1479 BSA molecules/RBC with a Kd of 1.91 (±0.09) × 10−7 M and a Bmax of 2.50 (±0.03) × 10−8 M, or 17,900 receptor molecules/RBC. At saturation, the additional 2,700 BSA molecules binding per RBC in the presence of C‐peptide and zinc indicates that an albumin receptor exists on RBCs as well as a separate albumin/C‐peptide complex receptor.Support or Funding InformationNIH
Role of astrocytes in cerebrovascular protection provided by an opioid agonist, biphalin during ischemic stroke
Authors: Nozohouri Saeideh , Albekairi Thamer , Zhang Yong , Vaidya Bhuvaneshwar , Abbruscato Thomas 
Published: April 2020

Biphalin, a dimeric enkephalin analog (Tyr‐D‐Ala‐Gly‐Phe‐NH‐)2 with potent, nonselective opioid receptor agonist activity, has shown to be neuroprotective in both in vitro as well as in vivo ischemic stroke models. Here we aimed to understand the role of biphalin on blood‐brain barrier (BBB) integrity during ischemic stroke. In this study, we used mouse brain endothelial cells, bEnd.3 and primary astrocytes as monoculture and co‐culture in‐vitro BBB models. Restrictive BBB properties were evaluated by measuring trans‐endothelial electrical resistance (TEER) and [14C] sucrose paracellular permeability, and the redistribution of the tight junction protein occludin. The protective effect of biphalin on BBB integrity was assessed after exposing cells to oxygen‐glucose deprivation (OGD) (1% O2, 2 hours) and glutamate (2mM). Naltrexone, a non‐selective opioid antagonist was used to determine opioid receptors dependent activity of biphalin. Further, we measured the effect of biphalin on astrocytic glutamate uptake and on the expression of excitatory amino acid transporter (EAAT) to study the indirect role of astrocytes on biphalin mediated BBB protection. It was found that combined stress (2mM glutamate and 2 hours OGD) significantly increased the [14C] sucrose paracellular permeability in bEnd.3 cells as well as in a co‐culture cell model. Biphalin (100 nM) treatment attenuated the effect of the combined stress which was reversed by the opioid antagonist naltrexone (100 nM), suggesting the role of opioid receptors in biphalin activity. It was also observed that combined stress (2mM glutamate and 2 hours OGD) condition, significantly reduced the glutamate uptake by astrocytes however, biphalin treatment increased glutamate uptake in primary astrocytes, suggesting the role of astrocytes, increased buffering capacity, in biphalin meditated protection of the BBB during ischemic stroke.Support or Funding InformationTexas Tech University Health Science Center
MAGEA9: Driver or Passenger in CARcinogenesis
Authors: Johnson Jodiene 
Published: April 2020

Type I Melanoma Antigen Genes (MAGEs) are a family of genes that are normally expressed in the spermatogonial cells of the testis and abnormally reactivated in a broad variety of cancers. Due to this expression pattern and the immune response they elicit when abnormally expressed, type I MAGEs are also known as Cancer Testis Antigens (CTAs). Melanoma Antigen Genes‐A (MAGE‐A) are CTAs whose expression in cancers have been correlated with poor patient prognosis. The biological function of several members of the MAGE‐A gene family in cancer remains unclear. Ectopic expression of type I MAGEA9 in normal cells results in increased cellular proliferation. We therefore hypothesize that MAGEA9 is needed for sustenance of proliferative phenotypes. Our goals are to elucidate the molecular mechanism by which MAGEA9 increases cellular proliferation and the mechanism(s) that regulate the expression of MAGEA9. We have employed RT‐qPCR array for cancer‐associated genes to determine signaling pathways modulated by expression of MAGEA9. Results from our RT‐qPCR array indicate that MAGEA9 expression causes an upregulation in the DNA mismatch repair genes MSH6 and MSH2 and the E3 ubiquitin ligase MDM2. To identify MAGEA9 binding partners we have used IP‐mass spectrometry, and our data indicate that MAGE‐A9 binding partners are involved in invasion. We will use a combination of in silico promoter analyses and chromatin immunoprecipitation (ChIP) to determine the transcription factor (s) regulating expression of MAGEA9. Collectively, these data provide a clearer understanding for the function of MAGE‐A9 in cancer. Elucidating the role of MAGEs in cancer may provide insight for the development of novel therapeutic interventions.Support or Funding InformationNSF Research Initiation Award to S.R. #HRD1764201Department of Education Award # P382G170104 (TITLE‐VII)
Usefulness of high‐fluorescence body fluid cell count (HF‐BF %) in Automated hematology analyzer SYSMEX XN‐350 for screening abnormal cells in body fluid analysis
Authors: Lim Jihyang 
Published: April 2020

BackgroundAutomated hematology analyzer Sysmex XN‐350 is operated by two analytical software, whole blood‐mode and body fluid mode. The body fluid mode can be counted the cells twice as many as than whole blood mode and it is suitable for body fluid analysis which the sample has low cell count. XN‐350 is calculated the cell counts including total nucleated cell, WBC, RBC, mononuclear(MN) cell, polymorphonuclea(PMN) leukocyte and differentiated MN%, PMN%, high‐fluorescence body fluid cell(HF‐BF)%. The cell types in HF‐BF are macrophages, mesothelial cells, synovial cells and other abnormal cells. Higher HF‐BF% means higher abnormal cells. We evaluated the correlation HF‐BF % by XN‐350 to microscopic examination.MethodsFive hundreds twelve body fluid samples enrolled in this study. All samples analyzed HF‐BF % by XN‐350 instrument and the results compared to microscopic examination. In preliminary study, fifty‐two body fluid samples analyzed by XN‐350. Twelve samples in all samples revealed less than 1% HF‐BF and they were all normal cells by microscopy, and then we decided the cut‐off level for manual slide review was above 1.0% HF‐BF. We analyzed abnormal cell detection rate according to HF‐BF %.ResultsIn five hundreds twelve samples, two hundreds eighteen samples(42.6%) were less than 1% HF‐BF and two hundreds nighty‐four samples(57.4%) were more than 1% HF‐BF. Twenty‐seven samples in more than 1% HF‐BF were detected clinically significant abnormal cells by microscopy. In according to the grade of HF‐BF %, 1.0~4.9% HF‐BF was one hundred thirty‐two samples including three samples(2.3%) that detected abnormal cells by microscopy, 5.0~9.9% HF‐BF was fifty‐one samples including six samples(11.8%) that detected abnormal cells by microscopy, 10.0~19.9% HF‐BF was fifty‐eight samples including six samples(10.3%) that detected abnormal cells by microscopy, more than 20.0% HF‐BF was fifty‐three samples including twelve samples that detected abnormal cells by microscopy. Therefore, higher HF‐BF % assumed higher detection rate of abnormal cells by microscopy.ConclusionHF‐BF % in SYSMEX XN‐350 instrument is useful for screening of abnormal cells in body fluid analysis. Cut‐off level of above 1% HF‐BF can be detected majority abnormal cells and decreased microscopic examination rate.
Differential effects of opioid versus alcohol dependence on pain avoidance behavior and regional endocannabinoid system deficiency
Authors: Cucinello Jessica Ann , Berner Melissa E. , Pahng Amanda R. , Edwards Kimberly N. , Edwards Scott 
Published: April 2020

In contrast to their analgesic effects, chronic use of opioids or alcohol often creates the paradoxical emergence of heightened pain sensitivity, termed hyperalgesia, during withdrawal. A hallmark symptom of hyperalgesia is the development of motivational strategies to avoid painful stimuli, and we have shown that adult male Wistar rats display pain avoidance behavior using the mechanical conflict avoidance system when in withdrawal from morphine (n=10–11/group) and alcohol (n=8–9/group). The present study seeks to identify how morphine and alcohol differentially affect pain avoidance behavior and endocannabinoid signaling within pain‐ and avoidance‐ associated brain regions, including the periaqueductal grey (PAG) and basolateral amygdala (BLA).Morphine dependence was induced via daily subcutaneous morphine injections (escalating dose of 10–20mg/kg) over 3 weeks. Alcohol dependence was induced via exposure to chronic intermittent ethanol vapor (CIEV) for 14 hours per day for 8 weeks. Pain avoidance behavior was quantified using the MCS as the latency for rats to exit a brightly lit start chamber onto nociceptive probes of varying heights (0–5mm) to enter a dark goal box. Western blotting was conducted for quantification of endocannabinoid system‐related proteins cannabinoid receptor 1 (CB1R) and eCB synthetic and catabolic enzymes diacylglycerol lipase alpha (DAGLa), monoacylglycerol lipase (MAGL).We found that morphine and alcohol dependence produced distinct patterns of pain avoidance behavior, where morphine dependence produced a hyperalgesic behavior and ethanol dependence produced an allodynic phenotype. This behavior negatively correlated with PAG levels of CB1R and DAGLa in morphine dependent animals and with the ratio of BLA DAGLa:MAGL in alcohol dependent animals, reflective of a decreased eCB tone in these animals. This suggests that exogenous cannabinoid treatment may be an effective therapy for substance‐induced hyperalgesia. Because substance‐induced hyperalgesia and negative affective components of pain are integral to addiction reinforcement, targeting these behaviors therapeutically could treat substance abuse.Support or Funding InformationNIAAA: T32AA007577, R00AA020839, R01AA025996, R21AA025736, NIH‐NICHD‐2018‐05
CblX is a New Cobalamin Syndrome Affecting Craniofacial Development
Authors: Achilleos Annita , Chern Tiffany , Tong Xuefei , Hill Matthew C. , Chaudhury Arindam , Reineke Lucas C. , Neilson Joel , Sun Jenny , Ray Russel S. , Watkins David , Thiagarajan Perumal , Dasgupta Swapan K. , James Martin F. , Poché Ross A. 
Published: April 2020

Combined methylmalonic acidemia and homocystinuria (cblC type), an inherited disorder of cobalamin (vitamin B12) metabolism, is a rare metabolic and multi‐systemic disease caused by mutations in MMACHC. Patients with cblC can have severe neurodevelopmental defects including microcephaly, hydrocephaly, and seizures as well as renal, cardiac and hematological defects. Recently, two new variants of cblC were discovered and termed cblX and cblX‐like. Rather than being due to mutations in MMACHC, cblX and cblX‐like result from homozygous mutations in the transcription cofactor HCFC1 and its transcription factor partner RONIN (THAP11), respectively. Patients with either HCFC1 or RONIN mutations were shown to have a dramatic reduction in MMACHC transcription. We have known that HCFC1 is an obligatory partner for RONIN and we have previously shown that the HCFC1/RONIN transcriptional complex directly regulates mouse Mmachc expression. These findings suggest that cblX and cblX‐like disorders comprise a novel family of rare and severe cblC‐like disorders that are transcriptional in nature. To better understand the cellular and molecular mechanisms underlying the pathophysiology of these devastating neurodevelopmental diseases, we have generated Hcfc1A115V and RoninF80L mice, which have the same point mutations observed in cblX and cblX‐like patients. Both Hcfc1Y/A115V hemizygous and RoninF80L/F80L homozygous mice exhibit defects in cobalamin metabolism consistent with an inherited vitamin B12 disorder, as well as defects that recapitulate those observed in the human cblX and cblX‐like patients, including severe brain developmental defects, cardiac malformations, and anemia. Additionally, these mice also exhibit craniofacial deformities, a developmental phenotype that has not been previously linked to vitamin B12. Our data identifies, for the first time, a role for RONIN/HCFC1 in craniofacial development. Together, the phenotypic and molecular data confirm that the RoninF80L and Hcfc1A115V mouse models will serve as powerful tools to further uncover the pathophysiology of this complex family of diseases, as well as to achieve a better understanding of the complexities of craniofacial development.Support or Funding InformationThis work is supported by R01 DE028298.
PGE2 is a Candidate Distal Nephron Remodeling Factor
Authors: Zapf Ava , Grimm P.R. , Al-Qusairi Lama , Delpire Eric , Welling Paul A. 
Published: April 2020

We recently discovered that DCT1 cell‐specific expression of constitutively active SPAK (CA‐SPAK) drives a remodeling process of the entire distal nephron that includes atrophy of the aldosterone‐sensitive distal nephron and parallel inhibition of ROMK and ENaC (Grimm et al, JASN ’17). The mechanism by which genetic activation of NCC drives downstream nephron remodeling remains elusive but observations that PGE2 inhibits ROMK and ENaC ex vivo raises that paracrine signaling may be involved (Jin et al, AJPR 2007; Guan et al, JCI 1998). Here, we tested the hypothesis that activation of NCC causes atrophy of the CNT through increased PGE2 synthesis and release.We found increased levels of PGE2 in kidney cortical homogenates of CA‐SPAK mice compared to control mice on a control diet. Feeding wild‐type mice (WT) a low potassium diet (LKD) to physiologically activate NCC resulted in a comparable elevation of PGE2 and ASDN remodeling response, characterized by a decrease in ROMK/ENaC. Administration of the NCC‐blocking diuretic, hydrochlorothiazide, to CA‐SPAK mice restored PGE2 to control levels but had no effect on PGE2 of control or SPAK KO mice. Thus, PGE2 is elevated in response to super activation of NCC.PGE2 synthesis is controlled by specific prostaglandin E synthase isoforms. As revealed in western blot analysis, the microsomal prostaglandin E synthase‐1 (mPGES1) was selectively elevated in the kidney cortex of CA‐SPAK mice and WT mice on LKD compared to control mice, and this response was blocked by thiazide diuretics.Localization of mPGES1 by quantitative microscopy surprisingly revealed that increased protein abundance of mPGES1 was confined to the CNT and CCD of CA‐SPAK or the DCT2 and CNT of LKD treated mice. No significant changes in mPGES1 were observed in the DCT1.In conclusion, these studies identify PGE2 as a possible remodeling factor that is released from the late DCT and ASDN in response to NCC activation. We speculate that decreased sodium delivery to the CNT and CCD when NCC is activated drives the response; similar to how low NaCl drives PGE2 synthesis in the macula densa. The paracrine/autocrine pathway provides a novel means to communicate potassium sensing in the DCT to potassium secretion in the ASDN by driving tubule remodeling of specific segments of the distal tubule.Support or Funding InformationNIH, NIDDK T32 and Fondation LeDucq
A putative GSK3β binding site in the aquaporin 1 (AQP1) C‐terminal tail controls pulmonary arterial smooth muscle cell (PASMC) function.
Authors: Yun Xin , Jiang Haiyang , Philips Nicolas , Shimoda Larissa 
Published: April 2020

Pulmonary hypertension (PH) is a devastating disease, characterized by functional and structural pulmonary vascular changes. In distal, small diameter pulmonary arteries, PASMC migration and proliferation are important contributors to the development of PH. In past studies, we showed that hypoxia, a stimulus for PH, elevated the expression of the water channel, aquaporin 1 (AQP1). We also showed that AQP1 induced PASMC migration and proliferation via a mechanism involving β‐catenin protein accumulation, which required the AQP1 C‐terminal tail but was independent of water transport. In this study, we wanted to elucidate the mechanism underlying AQP1‐mediated regulation of β‐catenin in PASMCs. β‐catenin can play various roles in the cell, and its protein levels are tightly regulated by the serine/threonine kinase, glycogen synthase kinase 3 β (GSK3β). Under control conditions, GSK3β is an important factor regulating the turnover of β‐catenin, binding to β‐catenin and targeting it for degradation. Recently, we performed in silico analysis of the AQP1 C‐terminal tail, and found in the proximal portion of the tail a potential binding site for GSK3β. Thus, we explored whether mutating the putative GSK3β binding site on the C‐terminal tail of AQP1 alters PASMC migration and proliferation and, if so, whether β‐catenin was involved. Primary cultured rat distal PASMCs were harvested and used for experiments. Cells were infected with adenoviral constructs expressing wild‐type AQP1 (AdAQP1), AQP1 with the putative GSK3β binding site mutated (AdAQP1M), or green fluorescent protein (AdGFP), as a control for infection. Infection with AdAQP1 and AdAQP1M increased total and surface AQP1 protein levels compared with AdGFP. The rate of change in water flux into PASMCs, tested by a water permeability assay using calcein self‐quenching, indicates that both AQP1 constructs function similarly as a water channel. While infection with AdAQP1 increased PASMC migration and proliferation, measured by BrdU incorporation and transwell assays, respectively, infection with AdAQP1M had no effect. Our results suggest that AQP1 controls PASMCs migration and proliferation via a mechanism that requires specific residues within the tail region that may form a putative GSK3β binding site. With increased AQP1 levels, as during PH, we think GSK3β is sequestered, β‐catenin escapes degradation, translocates to the nucleus and initiates transcription of the target genes involved in cell proliferation and survival.Support or Funding InformationThis work was funded by HL12651, 18POST34030262