bluuebunny/crossref_metadata_2025_split huggingface dataset:
Integrin‐targeting ferritin for drug delivery and tumor imaging
Authors: Chen Yan-Jun , Wang Hsu-Yuan , Huang Feng-Ting , Huang Chiun-Wei 
Published: April 2020

Various potent chemotherapeutic drugs have been developed over decades. Doxorubicin, the lead compound of the anthracycline family, has been applied clinically in a wide range of malignancies including leukemia, lymphomas, myeloma, breast cancer, lung cancer and sarcoma etc. Despite its profound therapeutic efficacy, this chemotherapeutic drug causes numerous side toxicity effects. One main side effect is cardiotoxicity. To eliminate the side effect of this conventional chemotherapeutic drug, targeted delivery of chemotherapeutic drugs to tumor cells is being investigated. This concept can be achieved using nano‐scaled drug‐containing particles, which are called nanoparticles. Many promising nanoparticles with various materials have been developed, such as liposomes, micelles, and carbon nanotubes. However, biocompatibility and metabolism issues related to these nanoparticles have limited their clinical applications. Hence, various nanoparticles have been fabricated to improve above disadvantages. Ferritin, consisting of 24 subunits, could self‐assemble into a shell‐like sphere and which make ferritin a suitable drug‐carrier. Moreover, with precisely assembled nanoscale shape, high safety and biocompatibility, ferritin has the potential for in vivo imaging and therapeutic purpose.Integrin α2β1 is a major collagen receptor that highly expresses on various tumor cells such as prostate, lung, and colorectal cancer cells. In this study, we have constructed integrin‐targeted ferritin which expresses integrin α2β1‐targeting peptide on the ferritin surface to specifically target cell lines with high‐expression of integrin α2β1. Our results indicated that the integrin‐targeted ferritin showed higher doxorubicin loading efficiency and yield than unmodified ferritin. Moreover, flow cytometry analysis showed that integrin‐targeted ferritin had better binding ability to U‐87 MG and PC3, cell lines with high integrin α2β1 expression, than unmodified ferritin. Furthermore, in cell cytotoxicity assay, doxorubicin encapsulated within integrin‐targeted ferritin showed higher cytotoxicity effects than doxorubicin encapsulated within unmodified ferritin and free doxorubicin in U‐87 MG and PC3 cells. All of our results support that integrin‐targeted ferritin has the potential to be a novel anticancer nanomedicine.Support or Funding InformationThis study was supported by Chang Gung Memorial Hospital, Taoyuan, Taipei, TAIWAN and Department of Biochemical Science and Technology, National Taiwan University, Taipei, TAIWAN
Exogenous Lactate Intake Can Induce Muscle Synthesis in Rats
Authors: Kyun Sunghwan , Yoo Choongsung , Jang Inkwon , Kim Jisu , Lim Kiwon 
Published: April 2020

PurposeLactate had been recognized as fatigue substance during high intensity exercise. However, the lactate paradigm has changed in the last decades, and it was reconsidered as a potential factors of energy resource and muscle adaptation. Nevertheless, the effect of administration of exogenous lactate on muscle has not been elucidated, and it has been no molecular signaling study of the muscle synthesis over time after ingestion. Therefore, we aimed to investigate the effect of lactate on muscle synthesis on skeletal muscle, according to the different time intervals within 2 hours.MethodsMale rats were randomly assigned into one of following four timings that were sacrificed time after oral administration of sodium lactate (2 g/kg); 0 min (n = 8), 30 min (n = 8), 60 min (n = 8) and 120 min (n = 8). After administration lactate, rats were sacrificed and immediately obtained blood and skeletal muscle (plantaris). To determine the effect of lactate on muscle synthesis (Akt, mTOR) and degradation (MuRF1, atrogin‐1), we measured mRNA expression in plantaris by RT‐PCR, and analyzed blood parameters of lactate, insulin, IGF1, glycerol and glucose.ResultsThe results showed that, despite no significant differences in the hormone receptors for androgen and IGF1 associated with synthesis, Akt remained 1.5~2 fold higher after 30minutes, and mTOR was lower than the 30‐minute expression level but showed a high trend at 60 and 120 minutes. In muscle degradation factors, MuRF1 remained elevated after 30 minutes, but atrogin‐1 had no change. Lactate intake increased plasma lactate by about 3.2 mmol / L at 60 minutes and decreased insulin and IGF 1 after 30 minutes. Furthermore, plasma glycerol was increased at 120 min, but glucose has not changed within 2 hours.ConclusionIt is demonstrated in the present study that lactate administration contributes to increase muscle synthesis factors at 30 minutes, and it remained at high levels within 2 hours. Moreover, the reduction of insulin and IGF1, and the upregulation lactate confirmed that the administration of lactate might be similar to the blood response of the human body during low or moderate exercise. Therefore, our results suggest that lactate intake as a supplement has the potential to increase muscle synthesis and have a positive effect on muscle injury and recovery after exercise. Despite these results, because our study is acute intake and synergies with exercise are not identified, further research is required to investigate the long‐term administration of lactate and the effects of exercise on muscle synthesis.Support or Funding InformationThis work was supported by the Ministry of Education of the Republic of Korea and the National Research Foundation of Korea(NRF‐2019S1A5A8033825)
Loss of von Hippel‐Lindau protein in renal collecting duct cells is associated with massive changes in gene expression and induces a mesenchymal phenotype
Authors: Edemir Bayram , Chernyakov Dmitry , Fischer Annika , Becker Franziska , Gallwitz Lisa , Bornkessel Nicola , Groß Alexander 
Published: April 2020

IntroductionClear‐cell renal cell carcinoma (ccRCC) is the most common form of kidney cancer. It is characterized by a loss of function mutation of the von‐Hippel‐Lindau tumor suppressor (VHL). The proximal tubule is traditionally the source of ccRCC but there is also evidence that a subset con also originate from collecting duct. Alongside the renal nephron, cells are exposed to increasing concentrations of NaCl and urea. In the collecting duct, osmolality can reach up to 1200 mosmol/kg, generating a hyperosmolar environment. It has previously been shown, that VHL deficient mice express a phenotype of polyuria and reduced aquaporin‐2 expression, which is predicted to be caused by a hypoosmolar environment in the nephron. This implicates a physiological function of VHL in the collecting duct segment. Most research however, that discusses the effects of VHL deletion has been conducted with renal cancer cell lines such as 786‐0, Caki or RENCA, which harbor various other specific mutations. Within this study we analyzed the impact of VHL deletion, using “healthy” renal collecting duct cells.Materials and methodsWe created a VHL deficient mpkCCD cell line by using CRISPR/Cas9. Single clones with functional VHL deletion were selected. The knockout was confirmed by western blotting. Cell morphology studies were performed, using the immune fluorescence method. The gene expression profiling was performed by Next Generation Sequencing analysis and results were validated with real time PCR. Proliferation and migration assays were performed for functional characterization of the cells.ResultsWe successfully generated single clones harboring dysfunctional VHL. This was associated with stable Hif1a expression. VHL deficient mpkCCD cells undergo drastic morphological and functional changes. We observed the partial loss of tight and adherent junctions. Deletion of VHL is also associated with higher migration and lower proliferation potential compared to wildtype cells. The expression of Aqp2, a specific marker of the collection duct epithelial cells, was massively reduced. RNA‐seq analysis showed massive changes in global gene expression pattern and cancer associated pathways were affected by VHL deletion. Correlation with the Humans Pathology Atlas showed that VHL loss induces an unfavorable gene expression pattern.ConclusionsSince the mpkCCD cells only lack active VHL, we are able to conclude a direct link between the loss of function of VHL and the cancerous transformation of epithelial cells. Therefore, our results might be useful for further research and novel treatment strategies in the future.Support or Funding InformationGerman Research Fundation
Racial Differences in Cardiometabolic Complications in Polycystic Ovary Syndrome
Authors: Stockton Savannah Marie , Alexander Nathan , Fernandez Edgar Torres , Huffman Alexandra , Reckelhoff Jane , Romero Damian , Cardozo Licy Yanes 
Published: April 2020

BackgroundPolycystic Ovary Syndrome (PCOS), the most common endocrine disorder in reproductive‐age women, is characterized by androgen excess and ovarian dysfunction. PCOS women have an increased prevalence of components of the metabolic syndrome, such as increased blood pressure (BP), insulin resistance and obesity. Whether African American women with PCOS have worse cardiometabolic profile than Caucasian remains unknown.AimsTo determine the prevalence of components of the metabolic syndrome in patients of different ethnicity with PCOS in the state of Mississippi.MethodsUniversity of Mississippi Medical Center (UMMC) de‐identified patient data from 2013–2017 was obtained from electronic medical records (EMR) using UMMC’s Research Data Warehouse. Cardiometabolic risk factors were analyzed and compared between African American and Caucasian PCOS women. Data is expressed as mean. SEM and statistical analysis was performed using SPSS.ResultsDuring the 2013 to 2017 period, EMR of 1,200 PCOS patients were obtained and analyzed from a total of 784,666 patients that were evaluated at UMMC. Compared to Caucasians, African American women with PCOS have significantly higher mean arterial BP (97.06±0.52 vs 94.23±0.48 mmHg p<0.001), hemoglobin A1C (6.03±0.08 vs 5.63±0.07 % p<0.001) and Body Mass Index (39.30±0.43 vs 36.05±0.39 p<0.001). About 44% of patients with PCOS have a BMI more than 40 (morbid obesity).ConclusionsMorbid obesity is highly prevalent in patients with PCOS in the Mississippian population cared for at UMMC. African American women with PCOS have a significantly higher prevalence of cardiometabolic risk factors than Caucasians. Lifestyle modifications should be implemented to ameliorate the prevalence of cardiometabolic risk factors in patients with PCOS.Support or Funding InformationFunding for this research is supported by NIH R01 HL135089 (JFR), P20 GM12334(LLY, ETF, JFR, DGR) and PO1HL51971 (JFR).
SPARC: Determine the topology and function of DRG neurons innervating mouse colon and rectum.
Authors: Feng Bin , Guo Tiantian , Bian Zichao , Chen Longtu , Zheng Guoan 
Published: April 2020

Visceral pain is the major complaint of patients with irritable bowel syndrome (IBS) that affects ~15% of the U.S. population, and satisfactory management of IBS pain remains an unmet clinical challenge. Neuromodulation targeting afferent somata in dorsal root ganglions (DRG) has emerged as a new and effective therapy for many types of chronic pain. To advance neuromodulation for treating IBS‐related visceral pain, new knowledge is needed regarding the topological distribution and neural encoding functions of lumbosacral DRG neurons that innervate distal colon and rectum (colorectum). Here, we implemented GCaMP6f to conduct high‐throughput optical recordings of colorectal afferent activities in lumbosacral DRG. By establishing this optical approach with single‐spike resolution, the current study characterizes the neural encoding from hundreds of colorectal afferents in the DRG, which are sparsely distributed and difficult to study with conventional methods. Using a mouse ex vivo preparation with distal colorectum and L5‐S1 DRG in continuity, we recorded 791 colorectal afferents’ responses to graded colorectal distension (15, 30, 40, 60 mmHg) and/or luminal shear flow (20–30 mL/min), then functionally classified them into four mechanosensitive classes, and determined the topological distribution of their somata in the DRG. Of the 791 colorectal afferents, 90.8% were in the L6 DRG, 8.3% in the S1 DRG, and only 0.9% in the L5 DRG. L6 afferents had all four classes: 29% mucosal, 18.4% muscular‐mucosal, 34% low‐threshold (LT) muscular and 18.2% high‐threshold (HT) muscular afferents. S1 afferents only had three classes: 19.7% mucosal, 34.8% LT muscular and 45.5% HT muscular afferents. All seven L5 afferents were HT muscular. In L6 DRG, somata of HT muscular afferents, i.e., putative colorectal mechano‐nociceptors were clustered in the caudal region whereas somata of the other classes did not cluster in specific regions. Outcomes of this study can directly inform the design and improvement of next‐generation neuromodulation devices that likely implement spatially selective DRG modulation to alleviate visceral pain in IBS patients.Support or Funding InformationSupported by NIH SPARC grant U01 NS113873 (BF and GZ)
Sex Differences in the Cerebrovascular Response to a Metabolic Stimulus
Authors: Pearson Andrew G. , Miller Kathleen B. , Howery Anna J. , Corkery Adam T. , Barnes Jill N. 
Published: April 2020

BackgroundBlood pressure control and regulation of cerebral blood flow are influenced by biological sex. We have previously demonstrated sex differences in cerebrovascular reactivity to hypercapnia in young adults. However, sex differences in the cerebrovascular response to a metabolic stimulus are unknown. As prostaglandins play a role in regulating cerebrovascular reactivity, evaluating cerebrovascular responses to a metabolic stimulus with cyclooxygenase (COX) inhibition may reveal important differences between males and females. Therefore, the purpose of this study was to investigate sex differences in the cerebrovascular response to a metabolic stimulus with and without COX inhibition in young adults.MethodsForty‐three young adults, 21 males (age = 28 ± 5 y) and 22 females (age = 25 ± 5 y), completed two levels of difficulty of the Stroop Color Word Test and the average response was calculated. Beat‐to‐beat mean arterial pressure (MAP) and middle cerebral artery velocity (MCAv) were measured at baseline and in response to each level of the Stroop test before and after administration of the COX inhibitor indomethacin. The peak MAP, MCAv, pulse pressure (PP), and cerebral pulsatility index (PI) responses were calculated as the highest 3‐beat average during each Stroop test.ResultsAt rest, MCAv was higher (Females: 65 ± 2, Males: 55 ± 2 cm/s, p < 0.01) and PI was lower (Females: 0.70 ± 0.02, Males: 0.78 ± 0.02, p < 0.05) in females compared to males, with no differences in MAP or PP. There were no differences in the percent change or peak MCAv, MAP, PP, or PI between males and females in response to the metabolic stimulus. COX inhibition reduced MCAv (Females: −28 ± 2%, Males: −28 ± 3%, p < 0.01 compared to baseline) and increased PI (Females: 17 ± 2%, Males: 20 ± 4%, p < 0.01 compared to baseline), with no change in MAP or PP. During COX inhibition, MCAv at rest was higher (Females: 46 ± 2, Males: 38 ± 1 cm/s, p < 0.01) and PI was lower (Females: 0.81 ± 0.03, Males: 0.93 ± 0.03, p < 0.05) in females compared to males, with no differences in MAP or PP. During COX inhibition, the percent change in MCAv, MAP, and PI in response to the metabolic stimulus did not differ between groups (p > 0.05); however, the percent change in PP during the metabolic stimulus was greater in males compared to females (Males: 16 ± 2%, Females: 9 ± 1%, p < 0.01).ConclusionYoung females had greater MCAv and lower PI at rest and this difference persisted following COX inhibition. Prior to COX inhibition, there were no differences between males and females in response to the metabolic stimulus. After COX inhibition, young males demonstrated a greater change in PP in response to the metabolic stimulus. These results suggest that COX inhibition reveals sex differences in the PP response to a metabolic stimulus.Support or Funding InformationFundingThis research was funded by NIH grant HL118154.
Maternal Nicotine Exposure During Lactation Alters Craniofacial Development
Authors: mohi amr , Kishinchand Rajiv , Durham Emily , Cray James 
Published: April 2020

Maternal exposure to nicotine has been linked to increased incidence of craniofacial abnormalities in infants. One of the craniofacial anomalies is craniosynostosis, which results from the premature fusion of the calvarial sutures. In the US, incidence of craniosynostosis is 1:1800–2500 births and often requires neurosurgical intervention to allow for normal brain growth. As the use of nicotine is increasing among young population specially through vape/electronic cigarettes use, we feel the need to investigate the effects of maternal nicotine on craniofacial development. We aim to determine the most critical period along pregnancy and lactation where maternal nicotine exposure could cause craniofacial anomalies and how does nicotine affect cranial sutures and other craniofacial structures. We compared the widths of coronal, sagittal, and posterior interfrontal sutures in offspring after maternal nicotine exposure during pregnancy only, lactation only, and pregnancy + lactation. Using Micro‐CT scans for cephalometric measurements, our preliminary data shows that maternal exposure to nicotine during lactation affects coronal suture development evidenced by decrease suture width and incidence of fusion in PN day 15 mice. We plan future experiments using histomorphometry and immunohistochemistry to better understand how sutural tissue and cells are affected by maternal nicotine exposure.Support or Funding InformationThis study was supported by research grants from the National Institute of Dental and Craniofacial Research[R03DE026192 (JC), F31DE026684 (ED)], National Institutes of Health National Center for Advancing Translational Sciences [UL1 TR000062]Plastic Surgery Foundation [Pilot Award 512114].
Store‐Operated Calcium Entry Upregulated IL6 Expression in Glomerular Mesangial Cells through NF‐κB Pathway
Authors: Chaudhari Sarika , Shotorbani Parisa Yazdizadeh , Wei Celine , Davis Mark E. , Ma Rong 
Published: April 2020

BackgroundIL6 is a pleotropic cytokine and functions as a pro‐ as well as anti‐inflammatory mechanism depending on the cell type or the specific pathological environment. It is unclear if a particular signaling pathway confers IL6 with this property. Immunological and inflammatory mechanisms play a significant role in the development of diabetic nephropathy (DN). Locally produced cytokines/chemokines contributes to the early changes and histological impairment in DN. Glomerular mesangial cell (MC) is a major cell type in glomerulus to produce cytokines/chemokines in response to diabetes and a major contributor to mesangial expansion in DN. We have previously demonstrated that the Orai‐1 mediated store‐operated calcium entry (SOCE) suppressed ECM protein production by MCs. The aim of this study was to determine whether and how SOCE in MCs regulated IL6 production by MCs.MethodsOrai1, the channel protein mediating SOCE in MCs was knocked down using the targeted nanoparticle‐siRNA delivery system in wild type C57BLKS/J mice at the age of 16 weeks. Immunohistochemistry was performed on the paraffin embedded kidney sections to examine glomerular IL6 expression. In cultured human MCs, the expression of IL6 was examined in culture media and whole cell lysates using ELISA and Western blot analysis, respectively in the presence of normal glucose (5 mM D‐glucose + 20 mM L‐glucose) with/without an activator (thapsigargin at 1 μM) or inhibitor (GSK‐7975A 10 μM) of SOCE for 15 hrs. To determine the role of NF‐κB in SOCE effects on IL6 production, human MCs were treated with an inhibitor of NF‐κB, helenaline or siRNA against p65 subunit of NF‐κB. Furthermore, the effect of SOCE on the nuclear translocation of p65 was examined by assessing abundance of nuclear p65 protein in response to activation and inhibition of SOCE.ResultsThe IL6 expression level was reduced in the glomeruli of the mice treated with nanoparticle/Orai1 siRNA for 2 weeks compared to that in the control mice. In cultured human MCs, thapsigargin significantly increased IL‐6 level in supernatant media and in whole cell lysates of MCs, and this effect was attenuated by GSK 7975‐A, a selective inhibitor of SOCE. However, tunicamycin, an ER stress inducer did not alter the amount of IL6 as thapsigargin did. Treatment with helenaline, an inhibitor of NF‐κB pathway, or knockdown of p65, a subunit of NF‐κB significantly blunted the thapsigargin‐induced increase in IL6 protein abundance. Moreover, thapsigargin stimulated the nuclear translocation of p65 in human MCs.ConclusionOrai1‐mediated SOCE positively regulates IL6 production by MCs through activation of NF‐κB pathway.Support or Funding InformationResearch Support: NIH/NIDDK (1RO1 DK115424); AHA Southwest Affiliate.
Identifying the 20‐HETE Binding Site on the 20‐HETE Receptor (GPR75)
Authors: Pascale Jonathan V. , Adebesin Adeniyi , Falck John R. , Schwartzman Michal , Garcia Victor 
Published: April 2020

The G‐protein coupled receptor GPR75 and its ligand the cytochrome P450 (CYP)‐derived vasoactive eicosanoid 20‐hydroxyeicosatetraenoic acid (20‐HETE), are implicated in the activation of pro‐inflammatory and hypertensive signaling cascades including decreased nitric oxide (NO) production and increased angiotensin converting enzyme (ACE) protein levels. Little is known as to how and where 20‐HETE interacts with GPR75. Computational modeling of the putative ligand binding pocket identified a site located across the fifth and sixth transmembrane on GPR75 (SiteScore: 1.182 and Dscore: 1.303). An independent analysis modeling the ligand‐receptor interaction between 20‐HETE and GPR75 places 20‐HETE in close proximity to the putative binding site location (Docking Score: −3.229, Glide Emodel: −29.045). Using the PRESTO‐TANGO methodology as a measure of receptor activation through the quantification of a ligand‐dependent β‐arrestin recruitment, 20‐HETE (1 nM) increased β‐arrestin recruitment by 3.48‐fold (± 0.63; p<0.05) compared to vehicle treated HTLA cells transfected with the WT‐GPR75‐TANGO construct. Recruitment of β‐arrestin was not observed with other structurally similar lipids including arachidonic acid, arachidonoyl ethanolamide and 12(S)‐HETE. Moreover, co‐treatment of WT GPR75‐TANGO cells with equimolar concentrations of 20‐HETE and the 20‐HETE antagonist N‐disodium succinate‐20‐hydroxyeicosa‐6(Z),15(Z)‐diencarboxamide (AAA) prevented the 20‐HETE‐mediated increase in GPR75 receptor activation, i.e., β‐arrestin recruitment [3.45 ± 0.19 vs. 0.89 ± 0.15 fold, 20‐HETE (1 nM) vs. 20‐HETE (1 nM) + AAA (1 nM), respectively; p<0.05]. Analysis of the 20‐HETE‐GPR75 interaction revealed several potentially key amino acids involved in hydrogen bond interactions with 20‐HETE including Ser205, Thr212 and Ser209. To assess the significance of these amino acids with respect to 20‐HETE‐mediated activation of GPR75, we generated site‐directed mutants of each residue. Cells transfected with the Ser205Ala and Ser219Ala mutant GPR75‐TANGO constructs exhibited similar increases in 20‐HETE receptor activation when treated with 1nM 20‐HETE (3.73 ± 0.79‐fold and 3.01 ± 0.57‐fold increases in luminescence, respectively, compared to vehicle treated controls; p<0.05). Interestingly, Thr212Asn GPR75‐TANGO transfected cells showed no significant changes to receptor activation when exposed to 20‐HETE (1nM) (1.00 ± 0.29 vs. 0.84 ± 0.18‐fold, vehicle vs. 20‐HETE). Taken together, this study identifies Thr212 as a critical residue of the 20‐HETE receptor binding site located across GPR75’s 5th and 6th transmembranes. Further studies are necessary to uncover other essential residues and their role in 20‐HETE binding and receptor activation.Support or Funding InformationThis study was supported by National Institute of Health grants HL39793 (M.L. Schwartzman), HL39793‐01S1 (V. Garcia) and the Robert A. Welch Foundation (I‐0011, J.R. Falck).