bluuebunny/crossref_metadata_2025_split huggingface dataset:
Day and Night Comparison to Maximize Learning in Sprague Dawley Rats (Rattus norvegicus)
Authors: Bagoyo Editha Angela , Drew Kelly , Jenkins Mackenzie 
Published: April 2020

Domesticated mice and laboratory rats have been traditionally viewed as nocturnal creatures, the vast majority of their daily activity occurring during the “dark” phase of their circadian cycle. Behavioral studies of rats have been traditionally performed during their “light” period when rats should be sleeping. The purpose of this study is to examine whether learning using a spatial, novel object recognition paradigm is enhanced if rats are trained and tested during the dark phase of their cycle rather than the light phase. 10 Sprague Dawley rats were placed in an open field apparatus and observed for 6 minutes at a time during their “light” phase. Measurements were made of distance traveled within the open field, speed of travel, time spent within different areas of the field, time spent mobile and immobile, time spent on grooming behavior, and time spent “rearing” (standing upright). Learning was assessed by observing differences in time spent with a novel toy and a familiar toy. All tests were repeated 1 week later during the rats’ “dark” phase, and results for the two phases were compared using a t‐test. Results showed no evidence of learning in either the light or dark phases, indicated by an increased time spent interacting with the non‐novel object compared to the novel object. Locomotor activity, total time spent interacting with both objects, and rearing behavior were significantly increased during the dark phase, while total distance traveled was significantly lower, which is likely related to the increased rearing. Though these results do not indicate enhanced learning in the rats during the dark phase, the observed differences in activity and behaviors compared with the light phase warrant further study as they could have implications for other behavior experiments performed on rats during each phase.Support or Funding InformationResearch reported in this publication was supported by the NIH Common Fund, through the Office of Strategic Coordination, Office of the NIH Director with the linked awards: TL4GM118992, RL5GM118990, UL1GM118991. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. UAF is an affirmative action/equal employment opportunity employer and educational institution:
www.alaska.edu/nondiscrimination.
The Effects of Christianity on Whole Body Donation within the African American Community
Authors: Triska Jeffrey Scott , Barremkala Malli , Farr A. Celeste 
Published: April 2020

INTRODUCTIONThe goal of this project is to understand if there is a relationship between Christian perspectives relative to the practice of whole body donation. Previous studies have pointed to a racial disparity among the rates of whole body donation between Caucasians and African Americans (AA). As a majority of AAs identify as Christians, there may be a connection between the religious beliefs and perspectives of body donation among individuals of the AA community. According to the Pew Research Center, 87% of this racial group claims a religious affiliation; 53% said they attend church at least once per week.METHODSAA adults were recruited from two majority AA Christian churches in Southeastern Michigan. Focus groups were held with 5–7 participants each. Five focus groups were conducted with a total of 26 participants between the ages of 21–80 (avg 48, median 51). Prior to the focus groups, participants answered a short survey on demographics and social characteristics including questions about how large of a role religion plays in their lives and how often they attend church. The focus groups were conducted for 1.5‐hours in a conversational manner to elucidate the participants’ thoughts, feelings, and perspectives on whole‐body donation.RESULTSThe majority of participants conflated the whole‐body and organ donation processes. They were generally unknowledgeable about whole‐body donation, but were supportive of the idea, stating that it was “beneficial for advancement and science knowledge,” and they “commend those who donate their body.” Several were knowledgeable about organ donation and said they were open to that option, but others expressed fears and concerns that healthcare professionals would allow people to die rather than provide life‐saving treatment if they knew they were a body donor. Christianity seemed to mostly be an encouraging factor for donating with participants stating “Scripture says we will get a new body, we are spiritual beings,” and “It’s [my body] on loan from him. It is being loaned to me.” Others seemed to be opposed to the process due to the necessity that the donor bodies be cremated after use. Most agreed that whole‐body donations are not religious convictions, but rather a matter of personal preference.CONCLUSIONOverall, the largest barriers to body donation seemed to be lack of knowledge and trust of the medical establishment. Christianity seems to mostly be an encouraging factor for donation. There should be more educational campaigns, particularly in AA churches to spread awareness of the process, methods, and benefits of whole body donation and its difference from organ donation. There needs to be more research done on this topic and a comparison to a group of non‐religious AAs to see if these effects hold.
Reproducibility of Whole‐Body Cooling Response Using Laser Doppler Flowmetry
Authors: Schwartz Kelsey S. , Shirazi Behnia R. , Lang James A. 
Published: April 2020

Laser Doppler Flowmetry (LDF) has been used as a non‐invasive tool to measure cutaneous microvascular function. While previous studies have assessed reproducibility with perturbations that increase skin blood flow, it is unclear as to the reproducibility of the whole body cooling response. We hypothesized that the reflex vasoconstriction response to whole‐body cooling would be reproducible. Eight healthy, young adults participated in two cooling bouts, each separated by 48 hours to one week. Participants donned a whole‐body suit and were cooled from a baseline skin temperature (Tsk) of 34°C to 30.5°C over a 30 min period. The final Tsk was maintained for an additional ten minutes. Mean Tsk was calculated as the unweighted average temperature of the calf, thigh, abdomen, upper chest, back, and upper arm. LDF flux was measured at the ventral forearm and cutaneous vascular conductance (CVC) was calculated at each 0.5°C reduction in skin temperature, CVC = flux/MAP and expressed as percent changes from baseline (%ΔCVC). The coefficient of variation (CV) for each 0.5°C reduction in skin temperature were determined (33.5°C = 44.3, 33.0°C = 27.9). The data indicate poor reproducibility at the onset of cooling. Reproducibility steadily improved with lower skin temperature ranging from moderate at 32.5°C to 30.5°C to good at the cooling plateau. The data suggest that skin blood flow measurements collected at colder skin temperatures are reproducible and may be used as a longitudinal indicator of microvasculature function.Support or Funding InformationThis study received funding from the Wilderness Medical Society Hackett‐Auerbach Grant, supported by the Academy of Wilderness Medicine(r).
Disrupting the protein signaling activity of MAPK pathway as target for metastatic colorectal cancer therapy
Authors: Nkembo Augustine T. , Lamango Nazarius S. , Jin Yonghoa , Tawfeeq Nada , Mensah-Mamfo Justin 
Published: April 2020

Despite recent advances including the use colonoscopy for early detection of colorectal cancer (CRC), the 2019 incidence and death rates projection for the disease in the U S were still alarming, as they stood at 145,600 and 51,020, respectively. Patients with the metastatic colorectal cancer (mCRC) subtype remain without hope as they do not respond or quickly develop resistance to the current available therapeutics. Over 50% of patients with CRC harbor constitutively active K‐Ras and 10% with B‐Raf mutations. Men of African American descent are more likely to be diagnosed with mCRC and will equally die of it compare to other races. Also, the most aggressive and deadly subtype with constitutively active mutant B‐Raf is commonly diagnosed in women and patients of more advanced age. Therefore, investigating novel targeted therapy to inhibit and disrupt the activities of the driver mCRC remains a matter of urgency. Recent tests for biological activities of novel agents, polyisoprenylated cysteinyl amide inhibitor (PCAIs) designed and synthesized to target the hyperactivity of aberrant monomeric G‐Proteins such as Ras and Rho protein family show promising results for continued development as a drug molecule. We hypothesized that the PCAIs would target and suppress the hyperactivity of aberrant Mutant K‐Ras and/or B‐Raf function that drive the aggressive mCRC subtype. To test this hypothesis, we screened for the potency of PCAIs on inhibiting the cell viability of 2D and 3D cultures of different CRC cells including Caco‐2, HCT116 and HT‐ 29 cells. We also evaluated the mechanism by which they induce cancer cell death. Our results indicate that the PCAIs NSL‐BA‐040 and NSL‐BA‐055 inhibited 2D Caco‐2 cells cultured at very low concentrations of 3.8 and 1.4, respectively, and 2.2 and 1.5 μM, respectively for HCT116 cells. For the 3D cultures, NSL‐BA‐040 and NSL‐BA‐055 induced cell death of spheroids at EC50 of 20 and 5.5 μM, respectively. NSL‐BA‐055 induced cell death by apoptosis, blocks spheroid growth and shrinks their volumes. The effects of the PCAIs on HT‐29 2D and 3D cell cultures were even more dramatic. The PCAIs tested here demonstrate significant potency on CRC cells while sparing HEK293 cells. Since these PCAIs demonstrate a strong drug‐like property at this early stage of their development, they would be further tested, characterized and hopefully developed as drug for CRC therapy, targeting the drivers of the metastatic subtype.Support or Funding Information“Research reported in this publication was supported by the National Institute on Minority Health and Health Disparities of the National Institutes of Health under Award Number U54MD007582. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health”.
Resveratrol Prevents Hyperoxia‐Induced Lung Inflammation
Authors: Reçica Shkëlzen , Kryeziu Islam , Thaçi Qëndrim , Kurshumliu Fisnik , Mladenov Mitko , Hadzi-Petrushev Nikola , Sopi Ramadan B. , Basholli-Salihu Mimoza 
Published: April 2020

Prolonged oxygen exposure of preterm infants results in the development of bronchopulmonary dysplasia (BPD), a chronic lung disease characterized with lung inflammation, remodeling and airway reactivity. Unfortunately, therapeutic options are limited. Resveratrol is known to have anti‐inflammatory and anti‐oxidant properties in different diseases, however it is not clear about its effects on BPD. In this study we investigated the effects of resveratrol in a rat pup model of BPD. We hypothesize that supplementation of rat pups with resveratrol prevents the lung inflammation induced by hyperoxia. At P5 rat pups were assigned either to room air (21% O2)‐ or hyperoxic (≥95% O2)‐groups and exposed for seven days. During this time, animals were supplemented daily with resveratrol (30 mg/kg of body weight; i.p.) or vehicle. At day P12 animals were euthanized and lungs were harvested, and processed for biochemical and immunohistochemistry studies. Pro‐inflammatory cytokines’ levels (IL‐1β and TNF‐α) were measured in whole lung homogenate using ELISA method. Immunostaining for T‐lymphocyte marker (CD3), B‐lymphocyte marker (CD20) and CD45 was performed on 5 μm lung sections. Levels of IL‐1β and TNF‐α in lung tissue of hyperoxic group of animals were increased significantly (p < 0.01) compared to room air group animals. Supplementation of hyperoxic animals prevented this increase of IL‐1β and TNF‐α, while did not change in room air animals. In hyperoxic animals there was an abundance of CD3, CD20 and CD45 stained cells in the lungs of rat pups exposed to hyperoxia compared to room air exposed pups. Supplementation of hyperoxic animals with resveratrol reduced the number of CD3, CD20 and CD45 stained cells. We conclude that resveratrol prevents lung inflammation induced by neonatal hyperoxia and we speculate that resveratrol might be an effective therapeutic approach to prevent the adverse effects of neonatal hyperoxia.Support or Funding InformationSupported by MEST
CNS Activity of an LHX2‐Associated Cis‐Regulatory Module
Authors: Zhang Michael , Cooper Allen M. , Britton Jessica C. , Pira Charmaine U. , Shin Alexandra , Oberg Kerby C. 
Published: April 2020

Limb development can be described along three axes – proximal‐distal, anterior‐posterior, and dorsal‐ventral. Fibroblast growth factors (FGFs) from the apical ectodermal ridge (AER) coordinate growth along the proximal‐distal axis, while sonic hedgehog (SHH), secreted from the zone of polarizing activity, directs anterior‐posterior patterning and expansion. AER‐related FGFs and SHH regulate each other in a positive feedback loop. This reciprocal loop is critical for proper limb patterning, however the molecular intermediates involved in this loop are only partially characterized. Previously, we identified LIM homeobox 2 (LHX2) as a downstream target of FGF in the FGF to SHH loop and determined that FGF upregulates LHX2 in the absence of de novo protein synthesis. This suggests that LHX2, expressed in the mesoderm subjacent to the AER, is a primary response target of FGF signaling.Cis‐regulatory modules (CRMs) within noncoding DNA can convey temporal and spatial regulation of an associated gene and are good candidates to convey the FGF‐mediated regulation of LHX2. We identified several potential CRMs (PCRMs) upstream of the LHX2 locus based on sequence conservation, active chromatin marks, and presence of transcription factor binding sites related to FGF signaling. Based on the number of active chromatin marks, we chose four of these PCRMs to investigate. We suspect that FGF regulates LHX2 through one or more of these PCRMs. Thus, we hypothesized that one or more of these PCRMs is active in the LHX2 expression domain during limb development.To test for activity of LHX2 PCRMs we used targeted regional electroporation (TREP) to introduce PCRM‐ptk‐GFP reporter constructs into the distal mesoderm of Hamburger‐Hamilton stage 23 (HH23) chicken limb buds. Transfection efficiency was determined by co‐transfection with a b‐actin promoter‐driven RFP construct. We determined PCRM activity by fluorescence microscopy, following 24 hours of incubation after TREP.None of the four PCRMs tested exhibited activity in the limb. However, subsequent TREP of the reporter construct containing PCRM ‐7 (~ 800 bp conserved region located 215 kb upstream of the LHX2 promoter) into the neural tube of HH10 embryos revealed green fluorescence at the midbrain‐forebrain boundary 24 hours post‐TREP. This activity suggests PCRM ‐7 may play a role in regulating LHX2 expression in the developing forebrain. Further studies are underway to correlate the activity of this CRM with expression of LHX2 or other genes in the topologically associated domain.
An ELK4 Binding Site is Critical for Activity of the LHX2‐Associated Sub‐AER Regulatory Module 1 (LASARM1)
Authors: Olafsen Christian Eric , Cooper Allen , Britton Jessica , Pira Charmaine U. , Oberg Kerby C. 
Published: April 2020

Fibroblast growth factors (FGFs) secreted from the apical ectodermal ridge (AER) coordinate limb growth and proximal‐distal patterning, while sonic hedgehog (SHH) from the zone of polarizing activity directs anterior‐posterior growth and patterning. Moreover, these signaling centers maintain each other’s expression in a reciprocal feedback loop. This regulatory loop coordinates limb patterning during outgrowth. However, the molecular intermediates involved in in this loop are only partially characterized. Previously, we identified LIM homeobox 2 (LHX2), expressed in the mesoderm subjacent to the AER, as a downstream target of FGF in the FGF to SHH arm of the feedback loop.LHX2‐associated Cis‐regulatory modules (CRMs) are reasonable candidates to convey the FGF‐mediated regulation of LHX2. We have identified a 1745 bp CRM located 483 kb upstream of the LHX2 promoter that is active in the sub‐AER region within the expression domain of LHX2. In silico analysis of this LHX2‐associated sub‐AER regulatory module 1 (LASARM1) revealed several FGF‐related transcription factor binding sites, including a site for ETS like‐4 (ELK4). We performed in situ hybridization for ELK4 mRNA and found a pattern coincident with LHX2 expression. Thus, we hypothesized that mutation of the ELK4 binding site within LASARM1 would disrupt FGF‐mediated enhancer activity.To identify whether the putative ELK4 transcription factor binding site contributes to LASARM1 activity, we performed site‐directed mutagenesis on the ELK4 binding site in the LASARM1‐ptk‐EGFP reporter construct. We used targeted regional electroporation (TREP) to introduce control and mutated reporter constructs into the distal mesoderm of Hamburger‐Hamilton stage 23 chicken limb buds. Transfection efficiency was determined by co‐transfection with a β‐actin promoter‐driven RFP construct. After 24 hours of incubation LASARM1 activity was determined by fluorescence microscopy.The fluorescence of LASARM1mutELK4 was markedly reduced relative to the LASARM1control. Our findings suggest that FGF signaling utilizes the ELK4 transcription factor to regulate LHX2 expression through the LASARM1 enhancer. Further work is needed to confirm this connection.
Metformin enhances nigrostriatal dopaminergic tone via activation of atypical protein kinase ζ
Authors: Choi Dong-Young 
Published: April 2020

Parkinson’s disease (PD) is the second most common neurodegenerative disease characterized by the progressive loss of the dopaminergic neurons in the substantia nigra leading to a decrease in the striatal dopamine. In our previous study, we demonstrated that metformin upregulated dopamine in the mouse brain, thus we designed this study to investigate the molecular mechanism behind such pharmacological effects. Here, we found that metformin enhanced the phosphorylation of tyrosine hydroxylase (TH) which was accompanied by increased expression of BDNF, GDNF, and activated their common signaling pathways in the nigrostriatal pathway. These effects were recapitulated in SH‐SY5Y cells suggesting a plausible role of neurotrophic factors in the activation of TH. Indeed, both BDNF and GDNF were essential for metformininduced TH activation. Importantly, we found that AMPK/aPKCζ/CREB pathway was required for metformin induced GDNF upregulation and TH activation. Our research is the first to reveal the critical role of DA‐upregulating effect of metformin via TH activation that critically requires neurotrophic factor induction. These results potentiate the candidacy of metformin not only as a neuroprotective agent but also as a restorative therapy for the treatment of PD.Support or Funding InformationThis research was supported by Basic Science Research Program through the Natonal Research Foundation of Korea funded by the Ministry of Education (2018R1D1A3B07049570).
MINER2.0 Combines ImageJ and R for Fast Nearest Neighbour Colocalization of 2D Multi‐Channel Fluorescence Images
Authors: Poburko Damon , dos Santos Anita , Jensen Gabrille , Kim BaRun , Pauls Andrew D. , Shalchy-Tabrizi Sophia , Ng Irvin 
Published: April 2020

IntroductionColocalization analysis has become a common tool of microscopic subcellular localization studies. There are many tools and packages available for colocalization analyses. MINER2.0 differs from many other packages by providing user‐friendly, highly customizable and scalable Nearest Neighbour analyses, drawing on principles of centroid localization used in single molecule imaging.Methods and ResultsTo circumvent issues related to colocalization indices that rely on (often) arbitrarily thresholding color channels, our analysis pipeline compares predefined sets of regions of interest (ROIs) or can perform a de novo search for the K‐Nearest Neighbours (KNN) around the predefined ROIs. The algorithm calculates and reports: puncta area, mean intensities, full‐width‐half‐max sizes and intensity, and distances between the centroid or perimeter of reference ROIs and the centers of ROIs in up to two comparator ROI sets. Users can define ROI centroids as center of mass, geometric center or the biaxial Gaussian fit of the puncta. This latter method can statistically detect differences in distances between centroids in separate image channels of 1.4 pixels. At higher magnifications this allows detection of distances separating centroids in separate color channels at <90 nm, below the diffraction limit of optical microscopy. The pipeline is written as a user‐friendly macro for ImageJ/Fiji to ease of access. New in version 2.0, we moved from a brute force KNN calculation in ImageJ with an O(n2) time complexity to a Kd‐tree calculation in R with O(nLog(n)) complexity. This reduces the KNN calculation for large ROI sets (i.e. >1000 ROIs) from hours in ImageJ to typically <10 s in R. This enables batched analysis of large image sets on standard desktop computers (e.g. 36,000 ROIs in six 2048 × 2048 pixel images analyzed in <1.5 hours). We discuss three use cases: (1) the analysis of anti‐colocalization of the vesicular nucleotide transporter with diverse vesicle and lysosomal markers in Neuro2A cells, (2) analysis of the decrease of mitochondria containing mitochondrial DNA in A7r5 cells treated with diverse stressors including angiotensin II, 5,6‐dideoxycytidine, rotenone and hydrogen peroxide, and (3) the prevalence of micronuclei as markers of genotoxic stress in A7r5 cells expressing loss‐of‐function variants of polymerase gamma that is responsible for replicating mitochondrial DNA.ConclusionMINER2.0 provides a user‐friendly interface for fast batch processing of spatial colocalization analyses with minimal subjective bias. It provides users with extensive output parameters to allow detailed understanding of the spatial relationships of fluorescently labelled structures within cells.Support or Funding InformationNatural Sciences and Engineering Research Council of Canada