bluuebunny/crossref_metadata_2025_split huggingface dataset:
Structural insights reveal the specific recognition of meiRNA by the Mei2 protein
Authors: Shen Siyuan , Jian Yanze , Cai Zhaokui , Li Fudong , Lv Mengqi , Liu Yongrui , Wu Jihui , Fu Chuanhai , Shi Yunyu 
Published: May 2022

Abstract
In the fission yeast Schizosaccharomyces pombe, Mei2, an RNA-binding protein essential for entry into meiosis, regulates meiosis initiation. Mei2 binds to a specific non-coding RNA species, meiRNA, and accumulates at the sme2 gene locus, which encodes meiRNA. Previous research has shown that the Mei2 C-terminal RNA recognition motif (RRM3) physically interacts with the meiRNA 5′ region in vitro and stimulates meiosis in vivo. However, the underlying mechanisms still remain elusive. We first employed an in vitro crosslinking and immunoprecipitation sequencing (CLIP–seq) assay and demonstrated a preference for U-rich motifs of meiRNA by Mei2 RRM3. We then solved the crystal structures of Mei2 RRM3 in the apo form and complex with an 8mer RNA fragment, derived from meiRNA, as detected by in vitro CLIP–seq. These results provide structural insights into the Mei2 RRM3–meiRNA complex and reveal that Mei2 RRM3 binds specifically to the UUC(U) sequence. Furthermore, a structure-based Mei2 mutation, Mei2F644A causes defective karyogamy, suggesting an essential role of the RNA-binding ability of Mei2 in regulating meiosis.
Public service motivation, role stress, and job burnout of police post-COVID-19: A study of the police in Taipei City in Taiwan
Authors: Chiu Jeng-Tzu , Chou Po-Pin , Wang Chun-Yuan 
Published: May 2022

AbstractAs the outbreak of COVID-19 has spread rapidly in Taiwan in 2021, police have more arduous tasks to fulfil. High-stress police workloads and long working times cause high job stresses that make them physically and mentally tired, eventually leading to job burnout (JB). Previous research refers that JB is caused by role stress (RS). Yet public service motivation (PSM) focuses on an individual’s service motivation to work in public organizations regarded as an inherent value of civil servants. This leads to our research questions as follows: Will police role stressors affect job burnout? How does public service motivation affect police stress and job burnout? This study conducted a questionnaire survey of frontline police in Taipei City, confirmed that job burnout is generally an extension of pressure caused by role stress. In addition, this study has shown public service motivation has a moderating effect on job burnout. Therefore, the government can provide more resources and benefits for the police to increase their motivation for public services, and indeed alleviate job burnout.
Roles for CCN proteins in Lysophosphatidic Acid and Sphingosine 1‐Phosphate Signaling in Prostate Cancer Cells
Authors: Balijepalli Pravita , Knode Brianna K. , Nahulu Samuel A. , Meier Kathryn E. 
Published: May 2022

IntroductionCysteine rich angiogenic factor (CCN1/Cyr61) and connective tissue growth factor (CCN2/CTGF) are members of the CCN family of matricellular proteins. These secreted proteins interact with extracellular matrix proteins and integrins to modulate cell signaling. CCN proteins have been implicated in survival and other mitogenic responses in various cancers. CCN1 and CCN2 are the two early inducible genes in this family. Lysophosphatidic acid (LPA) and sphingosine‐1‐phosphate (S1P) are lipid growth factors known to contribute to tumor progression in prostate cancer. In previous work, our group showed that LPA stimulates proliferation and migration in prostate cancer cells via the G protein‐coupled receptor LPAR1. Free fatty acid receptor 4 (FFAR4) agonists, inhibit LPA‐induced responses in prostate cancer cells. We also demonstrated that CCN1 expression is induced by LPA in human prostate cancer cells. However, the roles of CCN1 and CCN2 in cancer cell signaling have not been fully elucidated.HypothesisWe hypothesize that CCN proteins contribute to LPA‐ and S1P‐induced adhesion and mitogenic signaling in prostate cancer cells.Methods and ResultsPC‐3 human prostate cancer cells were used in the current study. For all experiments, cells were serum starved for 24 hours and then treated with and without 10 µM LPA or S1P. Immunoblotting showed that CCN1 and CCN2 were induced by LPA after 2‐5 hours in PC‐3 cells. The response was inhibited by the LPA receptor antagonist Ki1625. TUG‐891, an FFAR4 agonist, inhibited LPA‐induced CCN1 expression in PC‐3 cells. S1P also induced CCN1 expression in PC‐3 cells. In related experiments, whole cell extracts, and extracellular matrix were subjected to immunoblotting for CCN1, and immunofluorescence microscopy was used to localize the induced CCN1. CCN1 protein levels in extracellular matrix were increased 2‐5 hours after LPA treatment, as compared to untreated controls, and CCN1 was detected outside the cells by confocal microscopy. In cell adhesion assays, LPA‐treated PC‐3 cells exhibited an increase in adhesion to fibronectin‐coated 96‐well plates that was detected by 2 hours. Erk MAPK activation in response to LPA was biphasic, with the later phase occurring after CCN1/2 induction.ConclusionThe results of this study suggest that CCN proteins may play roles in LPA‐ and S1P‐induced signaling in prostate cancer cells and are thus potential therapeutic targets.
Post‐Pubertal Testosterone Implants Induce Hypertension in Female‐to‐Male Trans‐Sex Rat Model
Authors: Shawky Noha M. , Yanes Cardozo Licy , Reckelhoff Jane F. 
Published: May 2022

IntroductionAndrogens induce increases in blood pressure (BP) and risk of cardiovascular diseases (CVDs) in females, as seen in polycystic ovary syndrome females. However, it is not clear whether treatment with gender‐affirming hormone therapy in trans‐men have an impact on their BP or risk of CVD. Therefore, the present study was undertaken to test the hypothesis that chronic androgen supplements (starting post pubertal) in a rodent model of transgender men, will increase BP and promote metabolic dysfunction.MethodsFemale SD rats were implanted with testosterone propionate (TP, 7.5 mg in 5 mm silastic tubes, replaced every 3 wks, n = 10) starting at 7 wks of age (post‐puberty) and compared to controls (empty silastic implants, n = 10). Rats were weighed weekly. Radiotelemetry transmitters were implanted at 13 wks of age (n = 5‐6/group) and mean arterial pressure (MAP) was measured at 15, 21 and 25 wks of age. Body composition was determined at 15, 21 and 23 wks of age and proteinuria was determined at 15 and 21 wks of age (n = 4/group). Serum testosterone was measured by LC/MS at 15 wks of age (n = 4/group). Oral glucose tolerance test was performed at 11 and 22 wks of age (n = 4/group).ResultsTP supplements increased serum testosterone at 15 wks of age by 30‐fold compared to female controls (6.4 ± 1.5 vs 0.2 ± 0.05 ng/ml, respectively; p<0.005). Body weight (BW) increased significantly in TP group compared to controls by 10 wks of age (241 ± 4 g vs 220 ± 2 g, respectively; p<0.05) and was sustained through 26 wks of age (311 ± 7 g vs 276 ± 5 g, respectively; p<0.05). Fat mass and fat mass/BW were similar between TP and controls. Lean mass was significantly higher in TP group compared to controls at 15, 21 and 23 wks of age (254 ± 7, 268 ± 12 and 273 ± 12 g vs 218 ± 6, 227 ± 5 and 229 ± 5 g, respectively; p<0.01); however, when factored for BW, there were no differences between TP and controls. Proteinuria was significantly higher in TP group compared to controls at 15 wks of age (4.9 ± 1.2 vs 2.1 ± 0.3 mg/24 h, respectively, p<0.05) and 21 wks of age (7.6 ± 1.1 vs 2.4 ± 0.5 mg/24 h, respectively, p<0.005). Importantly, MAP was increased significantly in TP group at 15 wks of age (112 ± 3 vs 106 ± 4 mmHg, p<0.05), 21 wks of age (114 ± 3 vs 105 ± 2 mmHg; p<0.05) and 24 wks of age (115 ± 3 vs 104 ± 2 mmHg, p<0.05), compared to controls, respectively. Oral glucose tolerance test at 11 and 22 wks of age was similar between groups.ConclusionTestosterone used in a form of sex‐affirming therapy in female rats induced increases in BP that are independent of adiposity or Insulin resistance and maybe associated to renal injury. Future studies should determine whether these changes are sustained with aging, and the mechanisms responsible for the increase in BP in the female‐to‐male trans‐sex model.
Antimicrobial and cytotoxicity effect of gum Arabic‐silver nanoparticles
Authors: Ahmed Omnia A. K. , Ahmed Omnia A. K. 
Published: May 2022

ObjectiveThe objective of this study was to test the antimicrobial and cytotoxicity effect of silver nanoparticles (AgNPs) manufactured by means of green synthesis using gum Arabic against microorganisms associated with oral biofilm.MethodsTwo types of AgNPs (4g/1g) were synthesized from gum Arabic powder (GA). These GA‐AgNPs were characterized via UV‐Vis spectral analysis, DLS, FTIR, and TEM. Additionally, the stability of both GA‐AgNPs was tested in artificial media representing the condition on the oral cavity. Subsequently, their antimicrobial activity was tested against S.sanguinis, S.mutans, L.acidophilus,and C.albicansusing disc diffusion method. Their minimum inhibitory concentrations (MICs) were established. Finally, cytotoxicity was tested on the buccal mucosa fibroblast cells (BMF) using MTT assay.ResultsBoth GA‐AgNPs were spherical, ranging between 10‐20nm in diameter and were more stable in Mullier Hinton Broth compared to the other media tested. 4gGA‐AgNPs had relatively comparable zones of inhibition to chlorohexidine, whereas those for 1gAgNPs were negligible. The average MIC90 & MIC50 for 4gAgNPs range between 3.125‐12.5µg/ml, with 25‐100µg/ml for 1gGA‐AgNPs. The 1gGA‐AgNPs had more than 60% survival below 25µg/ml, while 4gGA‐AgNPs was below 3.125µg/ml.Conclusion4gGA‐AgNPs showed the highest antimicrobial activity against the tested oral microbes, however with higher cytotoxicity that needs to be reduced for further dental application.
Citronellol exerts lung protective and anti‐inflammatory effects through hampering NF‐κB in lipopolysaccharide‐induced acute lung injury in mice
Authors: Redha Samah M. , Kathem Sarmed H. 
Published: May 2022

BackgroundAcute lung injury (ALI) is one of the most serious conditions characterized by an exacerbation of inflammatory response in the lungs that can result from lung infection. Citronellol, a natural monoterpenoid found in citronella oil and other natural sources. In this study, the lung protective and anti‐inflammatory effects of citronellol were investigated in ALI induced by Lipopolysaccharide (LPS) in mice.Methodadult mice were allocated into groups, control mice received normal saline. Mice in the induction group received LPS 10mg/kg/day intraperitoneally and euthanized 2 hours later. Mice in the treatment groups received either 50mg or 100mg/kg/day of oral citronellol for 5 consecutive days prior to LPS injection.ResultsPretreatment with citronellol (50mg/kg/day) attenuated lung inflammatory events as observed from the significant reduction of TNF‐α (6.02±1.039 vs 29.20±4.77) and COX2 (4.12±0.23 vs 10.58±0.77) mRNA expressions in lung tissue compared to non‐treated mice. Interestingly, increasing the dose of citronellol to 100mg/kg/day also resulted in a significant reduction in both TNF‐α (3.30±0.745 vs 29.20±4.77) and COX2 (4.62±0.58 vs 10.58±0.77) mRNA expression compared to non‐treated mice.Looking for the upstream events revealed that NF‐κB gene expression was significantly hampered by citronellol 50mg/kg/day (5.11 ± 1.096 vs 11.88±1.57) and 100mg/kg/day (3.22± 0.253 vs 11.88±1.57) compared to non‐treated mice. Analysis of the data revealed that the effect of citronellol were dose dependent in terms of TNF‐α and NF‐κB gene expression.ConclusionThis study clearly revealed that citronellol exerted anti‐inflammatory and lung‐protective effects in LPS‐induced acute lung injury. The observed effects were dose‐independent and resulted through hampering of NF‐κB pathway.
A novel histogram based on the SEER database predicts overall survival in patients with high‐grade cerebellar glioma
Authors: Duan Hubin , Hao Chunyan , Zhang Yu , Qiao Binjun , Jing Zhijie , Chen Xianghui , Zheng Juan , Qiao Xiaohong 
Published: May 2022

ObjectiveHigh‐grade cerebellar gliomas are clinically challenging to treat, and there is a relative lack of large sample statistics as a prognostic criterion. In order to compare changes in patient survival under different conditions, we screened the risk factors that affecting survival in patients with high‐grade cerebellar gliomas, developed Nomograms and verified their accuracy.MethodsPatients with high‐grade cerebellar glioma from 1975 to 2016 were retrieved from the Surveillance, Epidemiology, and End Results (SEER) database, excluding patients with no biopsy or surgery, unknown tissue type, grade, or survival data, then coded patient data for classification and processed, and a prediction group and a validation group were randomly assigned using the CARET procedure in R (4.0.2). We use univariate and multivariate Cox regression models to screen variables that were significantly related to the prognosis of patients with high‐grade cerebellar glioma, and these variables were then used to construct Nomogram. Secondly, we use the consistency index (C‐index), receiver operating characteristic curve (ROC), calibration chart and decision curve analysis (DCA) to analytically validate the accuracy of the Nomogram model. Finally, Kaplan‐Meier survival analysis was used to test the prognostic indicators of Nomogram.ResultsA total of 197 patients with high‐grade cerebellar glioma were selected from the SEER database, and the patient data were categorized and coded into a prediction group (n = 100) and a validation group (n = 97). Univariate and multivariate Cox regression models were used to obtain four statistically significant indices of age, tumor type, type of surgery, and radiation therapy, which were then used to construct Nomogram with a C‐index of 0.830 (95% CI: 0.765‐0.895) for the prediction group and 0.789 (95% CI: 0.715‐0.863) for the validation group.ConclusionNomograms have promising clinical applicability and more accurate predictive outcome. For patients with cerebellar glioma, radiotherapy and surgical resection are definitive in improving prognosis. Older patients tend to die earlier, and the pathological type is also a key factor in prognosis.
Characterizing the Correlation Between Angiogenesis and Osteogenesis In Vivo Using Multicontrast Functional Imaging in a Calvarial Defect Model
Authors: Ren Yunke , Bhargava Akanksha , Senarathna Janaka , Chu Xinying , Grayson Warren , Pathak Arvind 
Published: May 2022

Angiogenesis is a key factor in bone healing that allows the delivery of oxygen, nutrients, inflammatory and bone precursor cells to the defect site. It induces dramatic structural and functional remodeling of the vasculature during the first 4 weeks of bone healing. Although recent advances in optical imaging have elucidated the in vivo relationship between angiogenesis and osteogenesis in a calvarial defect model, these efforts were mostly limited to structural imaging of the vasculature and bone. Therefore, to better characterize changes in vascular function (i.e. blood flow, oxygenation, etc.) during the bone healing cascade we developed a multicontrast optical imaging framework to assess in vivo changes in microvascular architecture using intrinsic optical signal (IOS) imaging; changes in blood flow with laser speckle contrast (LSC) imaging; and bone formation with bright‐field imaging at high spatial (5 µm) and temporal (200 ms) resolutions. With this system we acquired multicontrast images from 10 animals with calvarial defects every 2 days, over 4 weeks (Fig. 1a, d). The bone and vasculature were then digitally segmented from the bright‐field and IOS images for quantitative analysis (Fig. 1b, c). We also euthanized an animal each week for assessing 3D changes in bone volume and vascular architecture using an ex vivo, high‐resolution (10 µm) CT imaging workflow we recently developed called VascuViz (Fig. 2a, b, d). Using this framework, we demonstrated that angiogenic remodeling within the bone defect microenvironment was most robust from post injury day (D) 4 to D10, during which blood vessel length, volume and density all showed significant increase (Fig. 2c). The blood flow exhibited a sudden increase at D6 when angiogenesis started, and the angiogenic vessels were most perfused at D12 (Fig. 1d). These vascular changes peaked by the end of week 2 and plateaued during the next 2 weeks which were correlated with a large increase in bone volume during the first 2 weeks and smaller increase during the last 2 weeks (Fig. 2e). These results indicate that both structural and functional changes of the microvascular system strongly correlate with bone growth during the early stages of bone healing.Next, we plan to include additional indices of vascular function such as vessel maturity and intravascular oxygenation, as well as image‐based hemodynamic models to further characterize the role of the vascular microenvironment during osteogenesis. We believe that this novel imaging framework for characterizing the defect microenvironment can be utilized to inform the design of novel tissue engineering (TE) constructs for craniofacial bone regeneration. Collectively, these advances herald a new era of “image‐based TE”.
Elucidating the cellular mechanism of methionine toxicity
Authors: Pitsch Emily , Hughes Adam 
Published: May 2022

Cells possess several mechanisms to maintain amino acid homeostasis including regulation of amino acid permease trafficking, protein synthesis, metabolic reactions, and storage within the lysosome or vacuole. With aging and in certain diseases these processes are impaired, leading to a toxic accumulation of amino acids. While it is well‐established that excess amino acids are problematic for cells, it is poorly understood how individual amino acids drive cellular dysfunction and how cells combat toxic levels of these metabolites. To begin to elucidate cellular mechanisms of amino acid toxicity, we focused our analysis on methionine, because of its unique role in translation, metabolism, and as a precursor for the methyl donor, s‐adenosylmethionine. Additionally, methionine restriction has been shown to extend lifespan and delay progression or improve disease phenotypes, suggesting that methionine itself can be problematic to organisms.To study methionine toxicity, we first identified yeast strains that were sensitive to the amino acid. We genetically modulated pathways cells use to traffic two amino acid permeases – the high‐affinity methionine permease (Mup1) and the general amino acid permease (Gap1). These permeases are targeted for removal from the plasma membrane by ubiquitination of N‐terminal lysine residues, which is facilitated by an α‐arrestin protein. Mup1 internalization was inhibited by knocking out the α‐arrestin, LDB19, and Gap1 internalization was blocked usingGAP1K9R,K16R. Compared to wildtype, these mutants were greatly sensitized to exogenous methionine, measured by growth inhibition, highlighting the importance of modulating methionine levels to maintain cellular health. To explore mechanisms of methionine toxicity, we conducted RNA‐sequencing and found that excess methionine leads to an upregulation of genes under transcriptional regulation of Hsf1 and Gcn4. These transcription factors are activated in response to disruptions in protein homeostasis and tRNA charging, respectively. In addition to Hsf1 activation, we observed aggregation of heat shock proteins, Hsp104 and Hsp42, via fluorescent microscopy. Genetic depletion of Hsf1 using a tet‐repressible allele led to diminished growth on methionine compared to control cells, suggesting there is an increased requirement for Hsf1 when cells are exposed to excess methionine. Finally, we found a decrease in total protein when cells are exposed to excess methionine. This could be explained by the activation of Gcn4 which is activated in response to alternations in tRNA homeostasis and would reduce translation rates.The reasons for methionine toxicity are currently unknown, but our data suggests that excess methionine drives proteotoxicity and may perturb tRNA charging homeostasis. This work provides us with preliminary evidence for the mechanisms through which methionine imparts toxicity and how cells respond to excess levels of this amino acid.
Interleukin‐8 (IL‐8) as a Potential Mediator of an Association Between Trimethylamine N‐Oxide (TMAO) and Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) among African‐Americans at Risk for Cardiovascular Disease
Authors: Baginski Alyssa , Farmer Nicole , Baumer Yvonne , Wallen Gwenyth R. , Powell‐Wiley Tiffany 
Published: May 2022

Trimethylamine N‐Oxide (TMAO) is a microbial metabolite derived from the hepatic‐gut axis and linked to inflammation and cardiovascular disease (CVD). Studies have shown that TMAO is linked to CVD and its risk factor hyperlipidemia by enhanced macrophage foam cell formation or alterations in reverse cholesterol transport. Other mechanisms have not been reported to date. Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) is a largely hepatic expressed protein that blocks Low Density Lipoprotein (LDL) receptor recycling, leading to hyperlipidemia. The primary objective of this study was to investigate a potential relationship between TMAO and PCSK9 in order to explore a novel mechanism linking TMAO and CVD risk. We hypothesized that TMAO and PCSK9 would have a direct association among a sample of African American adults, a population at increased risk for CVD, and that this relationship would be mediated by inflammatory cytokines.African‐American adults at risk for CVD living in the Washington, DC area were recruited to participate in a cross‐sectional community‐based study that involved clinical examination and collection of fasting blood samples. Serum samples were analyzed for clinical CVD risk factors. Fasting levels of inflammatory cytokines (interleukin (IL)‐1 beta, tumor necrosis factor‐alpha, and interleukin‐8), TMAO, and PCSK9 were measured using Luminex and ELISA, respectively. Univariate and multivariate linear regression analyses and structural equation mediation analyses were conducted using STATA. All models were adjusted for body mass index (BMI) and atherosclerotic CVD risk score (ASCVD).The participant sample (n = 60) included a majority of self‐identified African‐American women (93.33%), who presented as obese (Mean BMI = 33). A significant association between TMAO and PCSK9 was identified in both unadjusted (β = 0.30, p = 0.02) and adjusted (β = 0.31, p =0.02) models. Both TMAO and PCSK9 were significantly associated with IL‐8 (TMAO: β = 0.45, p = 0.00; PCSK9: β = 0.23, p = 0.05) in adjusted models. Mediation analysis indicated that 34.77% of the relationship between TMAO and PCSK9 was explained by IL‐8 (Figure 1).Our findings indicate a potential PCSK9 involved pathway for TMAO and CVD risk. Furthermore, our results suggest this pathway is mediated by the inflammatory cytokine, IL‐8. Future studies may identify the role of dietary or stress‐related factors that could increase IL‐8 and potentially influence the TMAO‐PCKSK9 relationship.