bluuebunny/crossref_metadata_2025_split huggingface dataset:
The analysis of the transmission rate of locally administered siRNA to blood
Authors: Kokuryo Toshio , Yokoyama Yukihiro , Nagino Masato 
Published: June 2021

SiRNA is a promising therapeutic drug. Nek2 is a member of the serine/threonine kinase family. We previously reported the tumor suppressing effects of locally administered Nek2 siRNA (direct intra‐tumorous injection) in several cancer xenograft models. However, there is little information regarding to the pharmacokinetics of locally administered siRNA. This study sought to elucidate the transmission rate to blood and incidence of side effects after local administration of Nek2 siRNA in rat model. First, we mixed different amount of Nek2 siRNA with rat plasma (0, 1, 10, 100 and 500μM) and performed Northern blot analysis. Nek2 siRNA was detected in rat plasma in a concentration dependent manner. Next, the transmission of Nek2 siRNA to blood was evaluated after subcutaneous administration. Nek2 siRNA/biocollagen complexes were formed (500μM, 100μl) and were injected in femoral area. The blood was collected at different time point (5, 15, 30 and 60 min) and was analyzed by Northern blot. The detection of Nek2 siRNA in the blood was the highest at 15 min, and it disappeared within 1 h after the administration. There was no complication related to the local administration of Nek2 siRNA.Our data suggest that the local administration of siRNA is a safe delivery procedure with little cytotoxicity. However, further investigation will be needed for the future clinical application of siRNA.
Medical students applying newly learned anatomy knowledge as part of a mentoring and experiential learning environment with undergraduate and elementary students
Authors: Thang Christine Katie , Diaz Molly , Padilla Jessica , Ojukwu Kene , Lee Heajung , Schmalz Naomi , Wisco Jonathan 
Published: June 2021

Anatomy Academy (AA) demonstrates the need for anatomy medical education to engage, motivate, and support mentorship that reinforces medical education, provides pre‐medical support, and promotes scientific knowledge and healthy lifestyles. AA pairs medical students with undergraduate and elementary school students in a curriculum consisting of anatomy, physiology, health education, and physical activity to supplement the California 5th grade PE standards. Each team consists of one medical, one undergraduate, and 5–6 5th grade students. Medical and undergraduate students lead the teams in learning followed by physical activity outdoors. Pre‐ and post‐ AA self‐evaluations are conducted to assess knowledge and confidence. Medical and undergraduate students reported feeling more confident in their teaching skills and scientific knowledge. 5th graders responded with higher knowledge scores and interest in science. Medical students enjoyed applying their medical knowledge through teaching and mentoring. Undergraduates found value in being mentored. This curriculum and framework could be utilized in the future to develop mentorship programs among students at different stages in their education. AA was supported by the Albert Schweitzer Fellowship and the UCLA Monica Salinas Fellowship.Grant Funding Source: Albert Schweitzer Fellowship; UCLA Monica Salinas Fellowship
Insertion of a functional copy of six2 into the mouse genome to examine prenatal morphogenetic factors leading to chronic renal failure in the adult
Authors: Chang Cara , Stoytcheva Zoia , Marh Joel , Dee James , Fogelgren Ben , Lozanoff Scott , Moisyadi Stefan 
Published: June 2021

Suboptimal kidney development resulting in a decreased nephron number can have lifelong consequences and is associated with a significant risk of hypertension and end‐stage renal disease. The adult Br heterozygous mouse displays the symptoms of suboptimal kidney development along with a reduced six2 expression during embryonic renal morphogenesis. The purpose of this study was to determine whether a functional copy of six2 could be inserted into the Br mouse genome in an effort to rescue the mutant phenotype. A 26 KB six2 gene BAC fragment was isolated and processed by restriction digestion and pulse field gel electrophoresis. The resultant DNA fragment was then cloned into the mGENIE‐3‐BAC transposon vector via the Gibson Assembly method. The mGENIE‐3‐BAC‐six2 construct was subsequently confirmed by colony PCR, restriction digestion; the construct was also tested for functionality and expression in human cell lines and transgenic mice. Results from quantitative RT‐PCR demonstrated that a functional copy can be inserted into the human genome successfully, resulting in elevated transcription of mouse six2. Further experiments are being conducted to determine whether the adult CRF phenotype can be rescued prenatally by achieving an increased number of functional nephrons in adult transgenic mice.Grant Funding Source: R01‐DK‐064752 (SL), 5P20RR024206 and R01‐GM‐083158–01A1 (SM)
Study on the neurogenic mechanisms of traumatic brain injury
Authors: Duan Hubin , Fan Yimin , Hao Chunyan , Li Shuzhen , Wang Suping , Wang Hongqin , Kuai Dong , Lian Shizhong , Xue Naizhao 
Published: June 2021

Backgroud and purposeIn recent years, the mechanism of neurogenic inflammation in traumatic brain injury (TBI) gradually attracted people's attention. The purpose of research is to explore the specific mechanisms of traumatic brain edema caused by TBI.Methods40 male Wistar rats were randomly divided into control group, mild trauma group, moderate trauma and severe trauma group. Discharge frequency of the thalamic ventral posterior medial nucleus (VPM) were recorded after TBI.The expression of neuropeptide Y (NPY), substance P (SP), aquaporin‐4 (AQP4) and nuclear transcription factor‐kappa B (NF‐¦ÊB) were detected by immunohistochemical, Western blot, RT‐PCR and ELISA.ResultsWith the increase of trauma degree, the more obvious cerebral edema, the more visible neurotransmitters release and inflammatory cell infiltration between the synapses, the more gene and protein expression of SP, NPY, NF‐¦ÊB, and AQP4. There are positive correlations between the above gene and protein expression; There are also positive correlations between SP and the firing frequency of VPM.(P <0.05).ConclusionThe more severe damage of TBI, the more gene and protein expression of above neuropeptides. Neurogenic inflammation and neurogenic mechanisms play an important role in traumatic brain injury.
Regulation of the DNA‐binding protein CFP1 by ERK1/2
Authors: Klein Aileen M , Earnest Svetlana , Cobb Melanie H 
Published: June 2021

CFP1 (CXXC finger protein 1) was identified in our lab as an ERK2 interactor through a yeast two‐hybrid screen. CFP1 binds to the maintenance cytosine methyltransferase DNMT1 and is required for DNA methylation in an embryonic model. Methylation of DNA in promoter regions is critical for transcriptional silencing throughout development and is often dysregulated in cancer. ERK2 is required for KRAS‐driven aberrant gene silencing via promoter methylation by DNMT1, but ERK2 substrates that may mediate this effect have not been identified. I propose that CFP1 is the major ERK2 substrate that directs DNA methylation by promoting the targeting or activity of DNMT1. ERK2‐bound mononucleosomes are enriched for CFP1. Active ERK2 can phosphorylate CFP1 on multiple sites in vitro. Current work is focused on determining the requirement for CFP1 in KRAS‐driven gene silencing, and dissecting the relationship between promoter methylation and the presence of active ERK2 at DNA methylation sites guided by ERK2 ChIP‐sequencing data from a lung cancer model. If ERK1/2 targeting of CFP1 helps to drive directed DNMT1 activity and tumor suppressor gene silencing, it would cement a clear mechanistic linkage between a common activating mutation in cancer and conserved downstream epigenetic effects that promote development of the disease.This work was supported by NIGMS Training Grant T32 GM008203 and by NIH R37 DK34128.
LC‐MS plasma biomarkers associated with weight loss over 24 months following Roux‐en‐Y gastric bypass surgery
Authors: Bowers Jeremiah , LeGreve Tracy , Gletsu‐Miller Nana 
Published: June 2021

ObjectiveTo observe global changes in plasma metabolic profiles of patients following Roux‐en‐Y gastric bypass (RYGB) surgery.MethodPlasma global metabolite profiles from patients that were monitored before 6 months (n=13) and 24 months (n=5) following RYGB were determined using high performance liquid chromatography‐mass spectrometry (HPLC‐MS). Fat mass and dietary intake were assessed using air displacement plethysmography and 3‐day dietary records. Data analysis included repeated measures analysis of variance and Pearson correlations, (p < 0.05).ResultsDuring 6 months following surgery levels of 3‐indoleacetic acid and proline betaine increased while (9R,13R)‐1a,1b‐dihomo‐jasmonic acid, acetoacetic acid, L‐valine, and choline decreased. Over 24 months, DL‐2‐aminooctanoic acid increased. No cross‐sectional associations were found between metabolites and fat mass, dietary fat, dietary carbohydrate, dietary protein, or daily caloric intake.ConclusionsChanges in plasma metabolic profiles following RYGB suggest a metabolic response unrelated to changes in adiposity or macronutrient intake. The impact of specific foods will be investigated further.
Relationship between Inflammatory cytokines and bone mineral density in posmenopausal women with hormone replacement therapy.
Authors: Carvalho Cristina , Gonçalves Giovana , Bonetti Tatiana , Massad‐Costa Ana Maria , da Silva Ismael Guerreiro 
Published: June 2021

IntroductionBone formation and resorption are influenced by hormonal and inflammatory processes. Osteoporosis is a systemic disease condition with low bone mineral density (BMD), deterioration of the skeletal micro architecture and increased bone fragility highly prevalent at menopausal women.AimWe evaluated the relationships among inflammatory markers, hormone replacement therapy (E2) and bone mineral density (BMD) in healthy postmenopausal women and determined the contribution of inflammatory markers in BMD after 1‐year hormone replacement.Materials and MethodsThis preliminary analysis included 9 women evaluated pre and 1‐year pos E2 replacement. lumbar spine (LS) BMD were measured by dual energy x‐ray absorptiometry (DEXA) and serum inflammatory markers interleukin (IL)‐8, IL‐ 1β, IL‐6, IL‐10, IL‐12 and tumor necrosis factor‐alpha (TNF‐α) were measured at baseline and 1‐year post E2 replacement by flow cytometry using the Human Inflammation Kit BD Cytometric Bead Array (CBA). Statistical significance was analyzed by SPSS software and p<0.05 was considered statistically significant.ResultsThe Peason correlation analysis showed a significant negative correlation between IL‐1 β and BMD at baseline (r=− 0.803;p=0.009) and 1‐year (r=−0.768;p=0.016).ConclusionsIL‐ 1β is known for its role as inflammation mediator and recent researchers has shown involvement with ovarian processes. Our results suggest that increases in IL‐1β levels are related to decrease in BMD. Thun, modifying inflammatory markers, even in healthy postmenopausal women, may possibly reduce bone loss.
Stimulation of PAR2 during OIR reduces endothelial cell death and provokes rapid revascularization
Authors: Sitaras Nicholas , Joyal Jean‐Sebastien , Rivera Jose Carlos , Shao Zhuo , Sapieha Przemyslaw , Chemtob Sylvain 
Published: June 2021

In the retina, a highly regulated vascular meshwork irrigates neurons and supplies necessary metabolic requirements. Sustained vessel loss, as in Retinopathy of Prematurity (ROP), prompts revascularization despite initial suppression of vaso‐survival cues such as VEGF. Thus, other forces are likely involved. Importantly, we have previously demonstrated the substantial role of Protease‐activated receptor 2 (PAR2), a GPCR, in the developing retina. The exact mechanism by which PAR‐2 modulates angiogenic cues in ROP remains elusive.Expression and localization of PAR2 was determined by Western blot and immunohistochemistry in mouse whole retina. Appraisal of retinal vasculature and expression of pro‐ and anti‐angiogenic cues was assessed following intravitreal administration of PAR2 agonist peptide (SLIGRL) or lentiviral shRNA knockdown of PAR2 (LV shPAR2) in mice exposed to a murine oxygen‐induced retinopathy model (75% O2 from P7‐P12).Our results demonstrate a pronounced expression of PAR2 preferentially in RGC. PAR2 sharply increases during the initial phases of vaso‐obliteration (VO) and during the neovascularization (NV) phase. Activation of PAR2 with SLIGRL in vivo reduced Sema3A while increasing VEGF. Importantly, SLIGRL treatment in vivo prior to and following oxygen exposure significantly decreased VO and NV. Treatment with LV shPAR2 increased VO while augmenting NV.This study underscores the importance of the coupling between neurons and vessels and shows that RGC‐specific PAR2 plays a pivotal role in modulating angiogenesis in a pathological context.
Metformin modulates hyperglycemia‐induced endothelial dysfunction through SIRT1
Authors: Arunachalam Gnanapragasam , Samuel Samson M , Ding Hong , Triggle Christopher R 
Published: June 2021

Endothelial dysfunction, detected at an early stage in the development of diabetes‐related microvascular disease, is associated with accelerated endothelial senescence. Hyperglycemia‐induced endothelial senescence is an important contributing factor that promotes the development of ageing‐associated cardiovascular events. Clinical data indicate that the hypoglycemic agent, metformin, has an endothelial protective action. Our data with mouse microvascular endothelial cells cultured in high glucose (HG, 40mM), versus normal glucose (NG, 11mM), indicate a significant reduction in SIRT1 protein, an increase in FoxO‐1 and p53 acetylation along with an increase in the p21 levels and senescence‐associated beta‐galactosidase activity. Metformin treatment significantly abolishes the reduction of SIRT1 level and modulates the SIRT1 downstream targets FoxO‐1, p53/p21. We also investigated the role of small noncoding RNAs, miRNAs, as molecular targets that regulate SIRT1 expression. Our data demonstrates that hyperglycemia‐induced downregulation of SIRT1 plays a crucial role in diabetes‐induced endothelial senescence and, furthermore, the endothelial protective effect of metformin against HG‐induced endothelial dysfunction is, at least in part, due to its effects on SIRT1 expression and/or activity.This project was funded by grant # NPRP 4–910‐3–244 & NPRP 08–165‐3–054
Detection of the hyponitrite species (HO‐N=N‐O) in denitrification: Reactivity of NO with the heme Fe‐Cu center of cytochrome caa3 and the heme Fe –Fe center of Nitric oxide reductase
Authors: Varotsis Constantinos , Pinakoulaki Eftychia , Ohta Takehiro , Kitagawa Teizo 
Published: June 2021

Nitric oxide reductase (NOR) is the third of the four enzymes of bacterial denitrification that is responsible for the catalytic formation of laughing gas (N2O). Here we report on the formation of the hyponitrite (HO‐N=N‐O−) species (νN‐N=1332 cm−1) in the heme b3 Fe/non‐heme FeB dinuclear center of NOR from Paracoccus denitrificans. With the identification of the hyponitrite species the mechanism for the 2e−/2H+ reduction of NO to N2O can be described with more certainty.This work was financially supported by the Cyprus Research Promotion Foundation grant TEXNOLOGIA/THEPIS/0609(BE)/05.
TAZ Activation Drives Fibroproliferation in Spheroid Culture
Authors: Jorgenson Amy , Choi Kyoung , Sicard Delphine , Tschumperlin Daniel 
Published: June 2021

The transcriptional co‐activator TAZ plays a critical role in lung alveolarization (Mitani et al. Am J Respir Crit Care Med. 2009). We recently observed enhanced TAZ expression in the nuclei of fibroblasts present in lung tissue from patients with idiopathic pulmonary fibrosis, suggesting that TAZ is reactivated during fibrosis. To test the role of TAZ in driving a fibroproliferative response, we developed scaffold‐free cultures in which fibroblasts were grown in 3D hanging drop spheroids. This spheroid model removes the influence of the artificial 2D cell culture environment on cell growth. NIH3T3 fibroblasts were stably transduced with doxycycline (dox)‐inducible constructs expressing nuclear‐localizing mutant TAZ4SA or empty vector (courtesy of X. Varelas, Boston University). Spheroids were initiated by seeding 500 cells in hanging drops, and were subsequently fed growth media supplemented with 0, 1, 10, or 100 ng/mL dox every other day up to 15 days. Spheroid growth was tracked by measuring 2D projected area under light microscopy. Control NIH3T3 fibroblast spheroids remained stable in culture, but neither increased nor decreased in size significantly. In contrast, TAZ4SA‐transduced spheroids grew over time, and by day 15 spheroids treated with 100 ng/ml dox had a 2D projected area ~2.2‐fold larger than those grown in the absence of dox (p=0.0054). Intermediate growth levels were observed in 1 and 10 ng/ml doxycycline treated spheroids. Our results demonstrate the utility of the spheroid culture model for studying fibroproliferation, and document a prominent role for TAZ in promoting fibroblast spheroid growth.*Supported by NIH grants RO1 HL092961 and NIH R21 HL113796.
Comparison of LOCI Digoxin Assay and iDigoxin Assay for Rapid Detection of Convallatoxin, the Active Glycoside of the Lily of the Valley Plant
Authors: Everett Jamie , Kojima Yumi , Dasgupta Amitava 
Published: June 2021

ObjectiveLily of the Valley is a poisonous plant, used as an herbal remedy, with toxic effects due to convallatoxin, a cardiac glycoside. It has been reported that the LOCI digoxin assay, a luminescent oxygen channeling based digoxin immunoassay, has sufficient cross reactivity with Lily of the Valley extract, and the specific toxic component, convallatoxin. Yet, cross reactivity by the iDigoxin immunoassay, a chemiluminescent microparticle immunoassay, has not been reported in the literature. In this study, we evaluate iDigoxin immunoassay's ability to detect Lily of the Valley and convallatoxin, and compare these findings to those measured by the LOCI digoxin assay.Materials and MethodsAliquots of drug free and digoxin serum pools were supplemented with microliter amounts of Lily of the Valley extract or nanogram quantities of convallatoxin. Digoxin concentrations were measured using the iDigoxin immunoassay applied to the ARCHITECT i2000SR analyzer (Cabbott Laboratories), and the LOCI digoxin assay applied to the Vista 1500 analyzer (Siemens Diagnostics).ResultsThe iDigoxin assay measured significantly higher apparent digoxin concentrations than the LOCI digoxin assay in all supplemented aliquots. Also, the LOCI digoxin assay demonstrated bi‐directional interference with aliquots of a digoxin pool supplemented with various amounts of convallatoxin. However, with the iDigoxin assay all values were falsely elevated, indicating positive interference only.ConclusionsThe iDigoxin assay can be used for rapid detection of convallatoxin and Lily of the Valley in human serum. Also, the iDigoxin assay is more sensitive than the LOCI digoxin assay in convallatoxin detection.
Mechanism of Action in NaHS‐Induced Increase in Aqueous Humor Outflow
Authors: Robinson Jenaye , Okoro Esther , Mitchell Leah , Nguyen Olivia , Opere Catherine , Ohia Sunny , Njie‐Mbye Ya Fatou 
Published: June 2021

Previous studies from our laboratory demonstrate that the hydrogen sulfide (H2S) donor, sodium hydrosulfide (NaHS) can increase aqueous humor (AH) outflow facility in porcine trabecular meshwork tissues, ex vivo. In the present study, we investigated the mechanism/s that underlie the NaHS‐induced increase in AH outflow facilityMethodsPorcine ocular anterior segments explants were perfused with Dulbecco's Modified Eagle's Medium maintained at 37° C, 5% CO2 and constant pressure of 7.35 mmHg. Once outflow was stable (~3 hours), explants were exposed to the adenylyl cyclase inhibitor SQ22536 (100 µM), enzyme inhibitors for H2S biosynthesis [proparglyglycine (PAG, 1 mM); aminooxyacetic acid (AOA, 30 µM)], and prostanoids (flurbiprofen, 3 µM), 30 minutes prior to application of NaHS (10 µM). Outflow was then monitored for an additional 4 hours, and vehicle control (0.1% saline) was run in parallel.ResultsThe NaHS‐induced increase in AH outflow was attenuated by AOA and PAG by 79% and 56%, respectively. In the presence of flurbiprofen (3 µM), effects of NaHS on outflow was significantly (p<0.02) inhibited from [130.6 ± 10.51% of basal] to [102.9 ± 2.88% of basal]. Furthermore, the adenylyl cyclase inhibitor, SQ22536 completely blocked (p <0.05) the NaHS‐induced increase in outflow facility.ConclusionWe conclude that the ability of NaHS to increase AH outflow in porcine trabecular meshwork is dependent upon the intramural generation of H2S and prostanoids, and on the integrity of the adenylyl cyclase pathway.
Deletion of Soluble Epoxide Hydrolase Modulates Coronary Reactive Hyperemia in Isolated Mouse Heart
Authors: Hanif Ahmad , Zeldin Darryl , Nayeem Mohammed 
Published: June 2021

Coronary reactive hyperemia (RH) is impaired in hypertrophic hearts. Epoxyeicosatrienoic acids (EETs) exert cardioprotective effects in ischemia / reperfusion injury. Soluble epoxide hydrolase (sEH) is the main enzyme responsible for EETs breakdown. EETs level is increased in sEH knock‐out mice (sEH‐/‐). We hypothesized that in the absence of sEH, isolated mouse heart exhibits increased RH through increased generation of EETs in response to no‐flow ischemia compared to wild type (sEH+/+). Coronary flow (CF) in isolated sEH+/+ and sEH‐/‐ mouse heart was measured using Langendorff system. Perfused isolated heart was exposed to 15 second ischemia and RH was assessed. Following ischemia, flow repayment volume (RV) was significantly higher in sEH‐/‐ compared to sEH+/+ (7.6 ± 0.5 in sEH‐/‐ vs. 5.9 ± 0.7 in sEH+/+, p<0.05). Moreover, sEH‐/‐ exhibited significantly higher repayment to debt ratio when compared to sEH+/+ hearts (2.1 ± 0.2 in sEH‐/‐ vs. 1.5 ± 0.1 in sEH+/+, p<0.05). Upstream of EETs; the increased in RV in sEH‐/‐ was significantly decreased by SCH‐58261 (a selective adenosine A2A antagonist, 1.0 mM) (6.8 ± 0.8 in sEH‐/‐ without SCH‐58261 vs. 3.5 ± 0.3 with SCH‐58261, p<0.05). Also, downstream of EETs; T‐0070907 (a selective PPARg antagonist, 10.0 mM) significantly decreased RV in sEH‐/‐ (7.7 ± 1.2 in sEH‐/‐ without T‐0070907 vs. 5.4 ± 0.8 with T‐0070907, p<0.05).Our results demonstrate that deletion of sEH increases RH compared to WT and that the enhanced RH is significantly decreased by SCH‐58261 and T‐0070907. These data suggest that targeting or silencing sEH may have a beneficial effect on myocardial recovery from ischemia. Supported by HL‐114559 to MAN and z01‐ES025034 to DCZ