bluuebunny/crossref_metadata_2025_split huggingface dataset:
Oral Supplementation of Soybean And Hop Restored Bone Mass And Bone Turnover in Ovariectomized Rats
Authors: Lee Hansol , Noh Dasom , Lim Yeni , Kwon Oran 
Published: November 2023

A decline in the level of circulating estrogen induces reduction in bone mass and microarchitectural deterioration of bone tissue, placing postmenopausal women at a greater risk of osteoporosis. However, hormone replacement therapy (HRT) often provides the risk of oncogenicity and the adverse outcomes on cardiovascular disease. Thus, alternatively dietary intervention have been received a growing attention for their use with no accompanying adverse effects. The most significant body of evidence comes from the isoflavone flavonoids. Soybeans (Glycine max L.) and hops (Humulus lupulus L.) are the important sources of isoflavones, but provide different chemical profiles each other. Soybeans provides genistein and daizein, while hops provides prenylflavanone, isoxanthohumol, and xanthohumol. The aim of this study was to test whether a mixture of soybeans and hops (GH) alleviate bone changes found in ovariectomized rats. Eight‐week female Sprague‐Dawley rats were randomly divided into five groups (n = 10/group): sham‐operated control (CON), ovariectomized (OVX) control (O), OVX + 30 mg/kg BW of GH (OL), OVX + 100 mg/kg BW of GH (OM), and OVX + 300 mg/kg BW of GH (OH). After oral administration of each diet for 8 weeks, bone microarchitecture, bone turnover markers (osteocalcin, alkaline phosphatase, CTX, and NTX in plasma and urine), and osteoclastic and resorptive gene markers in the bone were measured. The GH supplementation mitigated the OVX‐induced increase in bone turnover rate and induced favorable architectural modifications of trabecular bone. Furthermore, GH suppressed osteoclastic gene expression as determined by RANKL/OPG ratio. The data suggest that the GH supplementation may be a promising alternative to HRT for the management of postmenopausal osteoporosis.Support or Funding InformationThis work was carried out with the support of “Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ01169502)” Rural Development Administration and the Bio‐Synergy Research Project (NRF‐2012M3A9C4048761) of the Ministry of Science, ICT and Future Planning through the National Research Foundation.
Persimmon Tannin Regulates Gene Expressions Important for Lipogenesis And Cholesterol Efflux in HepG2 Cells
Authors: Oh Eun Kyoung , Ahn Youngsook , Kwon Oran 
Published: November 2023

Persimmon (Diospyros Kaki) is a deciduous fruit and largely cultivated in the world, containing an array of bioactive compounds such as phenolic acids, tannin, flavonoids and dietary fiber. The health benefit of persimmon tannin has recently been reported in relation to the hypocholesterolemic, hypolipidemic and antioxidant properties in animals. However, the regulatory mechanisms have yet not to be elucidated. Here, we tested the hypothesis that persimmon tannin exerts inhibitory effects against the accumulation of both triglycerides and cholesterol in fatty‐acid challenged HepG2 cells. Tannin‐enriched persimmon extract (PT) was prepared by pressurized water extraction (121°C, 90 min), filtration, and concentration (20 mmHg, 125 rpm, 30°C). Tannin concentration of PT was enriched by 20 times compared with that in the persimmon powder. HepG2 cells were treated with fatty acid in the presence of PT at 0, 0.25, 0.5, and 1 mg/mL. Inhibitory effect of PT on lipid accumulation was confirmed by Oil‐Red‐O staining. Gene expressions related to triglyceride synthesis (fatty acid synthase, FAS; and sterol regulatory element binding protein‐2, SREBP‐2), cholesterol synthesis and excretion (LDL receptor, LDLR; proprotein convertase subtilisin kexin 9, PCSK9; 3‐Hydroxy‐3‐methylglutaryl CoA reductase, HMGCR; and ATP‐cassette binding proteins G5/8, ABCG5/8), and cholesterol efflux via bile acid synthesis and excretion (cholesterol 7‐alpha‐hydroxylase, CYP7A1; and bile salt export pump, BSEP) were measured by real‐time quantitative PCR. The results revealed significant decreases either in lipogenesis (FAS and SREBP‐2) or in cholesterol synthesis (HMGCR and PCSK9) and a significantly decrease in cholesterol excretion transporters gene (ABCG5, ABCG8) expression, whereas a significant increase in cholesterol uptake and efflux (LDLR, CYP7A1 and BSEP). These results suggest that PT may have a potential to modulate both lipid and cholesterol metabolism. The physiological relevance test of this in vitro observation is ongoing in rats treated with a high fat and cholesterol diet.Support or Funding InformationThis work was carried out with the support of Cooperative Research Program for Agriculture Science & Technology Development Rural Development Administration (PJ01169502) and the Bio‐Synergy Research Project (NRF‐2012M3A9C4048761) of the Ministry of Science, ICT and Future Planning through the National Research Foundation.
A Novel Assay for Cardiolipin Quantification in Isolated Mitochondria and Cell Lysates
Authors: Bahl Vasundhra , Saddar Sonika 
Published: November 2023

Cardiolipin is a phospholipid present in the inner mitochondrial membrane. It plays an essential role in regulating the enzymes involved in mitochondrial respiration and processes such as biogenesis, fission, fusion, protein transport and apoptosis. Diabetes, heart failure and aging are linked to changes in levels of cardiolipin. Exposure to toxicants like cigarette smoke and organophosphates causes an alteration in cardiolipin levels and composition, and adversely affect health. Cardiolipin abnormalities are associated with various diseases such as Barth's syndrome, a genetic disorder affecting the gene responsible for cardiolipin synthesis that can lead to cardiomyopathy, underdeveloped muscles and growth delay. Since cardiolipin is crucial to mitochondrial functions there is increasing interest in assays for determining levels of cardiolipin in biological samples.Until now, cardiolipin has been detected using mass spectrometry which requires very sophisticated instrumentation and expertise in the technique. BioVision's Cardiolipin Quantification Assay is a simple, straight‐forward and versatile plate‐based fluorometric assay that uses a proprietary probe. Our assay is highly specific for cardiolipin and no cross‐reactivity is observed with other lipids such as phosphatidylcholine, phosphatidylserine and sphingomyelin. The kit can detect as low as 0.2 nmol of cardiolipin in isolated mitochondria as well as cell lysates.
HIF‐1a‐Mediated Induction of Insig‐2 Regulates Cholesterol Metabolism by Accelerating Degradation of HMGCR
Authors: Hwang Seonghwan , Jo Youngah , Wolff Nicholas , Brugarolas James , DeBose‐Boyd Russell 
Published: November 2023

HMG CoA reductase (HMGCR) catalyzes a rate‐limiting reaction in cholesterol synthesis. In cultured cells, sterols exert tight feedback control on HMGCR by inhibiting its gene transcription and by accelerating its proteasomal degradation. Both of these processes require endoplasmic reticulum membrane proteins called Insig‐1 and Insig‐2. Although the transcriptional component of the HMGCR regulatory system has been scrutinized in livers of mice through various transgenic and knockout animals, regulated degradation of HMGCR in animal livers has not been studied in detail. HMGCR protein accumulates disproportionately to its mRNA in livers of mice genetically deficient in both Insigs, suggesting that Insig‐mediated degradation significantly contributes to feedback regulation of the enzyme. As cholesterol synthesis is an oxygen‐intensive process, the current study is aimed at elucidating whether hypoxia modulates Insig‐dependent degradation of HMGCR in liver of mice.To determine if hypoxia affects Insig‐dependent degradation of HMGCR in vivo, a liver‐specific transgenic mouse model was developed which expresses a T7‐tagged membrane domain of HMGCR, which is necessary and sufficient for degradation of HMGCR. As the transcription of the transgene is driven by a constitutive apoE promoter, the feedback regulation of the transgene product is exclusively dependent on proteasomal degradation. In this mouse model, we demonstrated that mice exposed to hypoxia stabilized hypoxia‐inducible factor (HIF)‐1a, increased Insig‐2 mRNA and protein, and accelerated degradation of HMGCR. Similar results were observed in mice administered with dimethyloxalylglycine (DMOG), a pharmacological inhibitor of prolyl hydroxylase that mimics hypoxia. DMOG failed to reduce the levels of HMGCR protein in mice that express the mutant HMGCR which is resistant to Insig‐induced ubiquitination, indicating that degradation of the enzyme significantly contributes to DMOG‐induced inhibition of HMGCR.In cultured human cell lines, DMOG treatment stabilized HIF‐1a, up‐regulated Insig‐2, and accelerated degradation of HMGCR in a dose‐dependent manner. RNAi‐mediated knockdown of HIF‐1a, but not of HIF‐2a, impaired DMOG‐induced expression of Insig‐2 and degradation of HMGCR. Moreover, DMOG‐induced degradation of HMGCR was blocked in cells lacking Insig‐2, whereas DMOG continued to augment HMGCR degradation in Insig‐1 deficient cells. Using chromatin immunoprecipitation and luciferase promoter assays, we show that Insig‐2 is a target gene of HIF‐1 in human and mouse cells. This is consistent with the presence of a putative HRE in the promoters of the human and mouse Insig‐2 gene. Overall, these findings suggest that Insig‐2 is a direct transcriptional target of HIF‐1 and that hypoxia‐induced degradation of HMGCR is mediated through HIF‐1a stabilization and subsequent up‐regulation of Insig‐2.Support or Funding InformationNIH Grants HL20948, GM112409
A Novel Yeast‐Based Screening System Identifies Signal Motifs that Regulate Membrane Protein Trafficking
Authors: Bernstein Joshua , Okamoto Yukari , Shikano Sojin 
Published: November 2023

The cell surface density of membrane proteins dictates their corresponding cellular activity. Many diseases, such as cystic fibrosis, arise from defects in cell surface trafficking of membrane proteins. Intracellular trafficking of membrane proteins is often controlled by the distinct signal motifs on the cargo proteins. A systematic approach to identify trafficking signals would facilitate our understanding of membrane protein trafficking. Here we report development of a new yeast‐based screening system that enables identification of the sequence motifs that down‐regulate surface expression of membrane proteins. We show that B31, a mutant S. cerevisiae strain lacking potassium (K+) efflux transporters, is unable to grow in high K+ media when expressing the mammalian K+ uptake channel Kir2.1 due to overloaded K+. However, B31 expressing the Kir2.1 channel defective in post‐Golgi trafficking or those fused with intracellular targeting motifs were rescued in high K+ media. Thus a growth rescue of B31 cells in high K+ media represents the reduction in the surface expression of the reporter Kir2.1 protein. Using this system, we have developed a method of screening a random C‐terminal peptide library fused to Kir2.1 channel for signals which reduce the levels of Kir2.1 on the cell surface. We have conducted a pilot screening, and identified sequences that down‐regulated cell surface expression of Kir2.1. We demonstrate that the identified sequences are functionally transplantable into different protein contexts such as CD4 and CD8. Furthermore, we have characterized the molecular mechanism of transport caused by these sequences and show that the screening system can identify signal sequences that operate through multiple distinct trafficking pathways such as ER retrieval and endocytosis. Thus our system offers a unique tool to discover potentially novel sequence motifs that regulate cell surface expression of membrane proteinsSupport or Funding InformationNIH R01GM099974
Drug Delivery in the Eye
Authors: Hosten Justin , Penumutchu Swathi , Lavik Erin 
Published: November 2023

On average, 1.4 million Americans suffer a head related injury every year. Of these injuries many can result in eye injury, in particular retinitis pigmentosa(RP). Recent innovation has lead to the creation of biodegradable microspheres and nanoparticles. One such use of these spheres/particles is to be a carrier for various drugs and be used for delivery to the human body, where oral drugs fail. For optic nerves, the blood brain barrier, which protects the brain from various chemicals that could have major negative effects, blocks many oral drugs from reaching the area where they are located. This poses a problem from optic related injuries. The drug All‐Trans Retinoic Acid (ATRA), is a compound that has the ability to repair damaged optic nerves. This drug can be loaded and directed to the optic nerves through poly (lactic‐co‐glycolic acid) (PLGA) microspheres. PLGA is a biodegradable polymer that allow for control and targeted release for various drugs. We hypothesize that loading ATRA with the addition of a co solvent, trifluoroethanol (TFE), methanol, and acetone, will increased the amount of ATRA within the microspheres, due to the hydrophilic properties of ATRA. This increased loading of the drug will allow for sustained release over long person of time, improve patient compliance, and patient treatment.Support or Funding InformationFunded by NIH and Glaucoma Foundation
Localization of the Platelet‐Activating Factor Receptor on the Intact Human Periodontal Ligament
Authors: Cerutis D. Roselyn , Nichols Michael G. , Khan Shakeel A. , Miyamoto Takanari 
Published: November 2023

Our laboratory has reported the presence of the lysophosphatidic acid receptor subtypes (LPARs) LPA1 and LPA3 on the intact human periodontal ligament (huPDL) using an in situ technique we developed (Cerutis et al. 2016). PAF (platelet‐activating factor) is another lipid mediator that also plays key homeostatic and inflammatory roles. Like LPA, it acts at nM concentrations and is found in significantly higher concentrations in saliva and gingival crevicular fluid from patients with chronic periodontal disease (PD), as we have reported for LPA (Bathena et al. 2011). It is now established that members of the LPA, PAF and prostaglandin (EP) GPCRs can localize to the nucleus in mammals (Gobeil et al., 2003).The LPAR and PAFR GPCRs are expressed in nuclear membranes in other systems, where they share signal transduction pathways to activate inflammatory gene transcription like COX‐2 and inducible nitric oxide synthase. Therefore, we hypothesized that the PAFR would also be expressed throughout healthy and PD PDL, where they would likely interact with LPARs to regulate the inflammatory response to periodontal pathogens.MethodsPolyclonal antibodies (Abs) against LPA3 and PAFR were covalently tagged with R‐Phycoerythrin and used to label both receptors on PDL from healthy and PD patients (IRB‐approved, consented donors), using our in situ technique.ResultsConfocal microscopy confirmed nuclear LPA3, and nuclear labeling was detected with PAFR Abs; similar but different patterns of distribution were seen for each GPCR. In conclusion, to the best of our knowledge, this is the first report of the presence of the PAFR on intact huPDL. In inflamed periodontal tissue, PAF may play a significant role, as it induces vasodilation and platelet aggregation; LPA is produced by activated platelets, so they are likely to interact at the molecular level. We reported that LPA regulates the expression of >60 PD‐associated inflammatory transcripts in human oral fibroblasts (Cerutis et al. 2015), supporting LPARs as having a regulatory role in chronic PD. This confirmed presence of PAFRs on the huPDL suggests that further investigation of the interplay of these two receptor families is needed to determine their role in both PDL homeostasis and in chronic PD.
Fix the Wrecks, RecA
Authors: Dubiel Katarzyna , Liang Adream , Sandholm Lily , Keck James L 
Published: November 2023

Accurate and complete DNA replication is essential to all life. However, collisions and obstacles that lead to DNA breaks are common. There are many proteins that have the essential job of correcting these mistakes. Some do this by trimming the broken ends of the strands and reconnecting them. RecA instead assists a more accurate method of repair called homologous replication. In this process, RecA binds to the broken strand of DNA and locates the same sequence on an intact duplicate strand. The ssDNA bound RecA then catalyzes a synapsis reaction between the filament and homologous double stranded DNA (dsDNA). Other proteins add complementary bases to synthesize a second copy of the missing section of the original single stranded DNA (ssDNA). Finally, the strands are separated to form a complete DNA and restore the missing information (Cox, MM. (2007)).Homologs of RecA are found in almost all organisms and are a critical part of genome maintenance and DNA repair. It is crucial for accurate homologous recombination which, when done incorrectly can lead to cancer and genome instability (Chen, Z., et al (2008)). When Rad51, the human homolog of RecA is dysregulated, it increases chances of cancer. Specifically, Rad51 is underexpressed in 100% of renal cell carcinomas (Liu, S., et al (2016)). Understanding the implications of cell repair regulation may allow for future cancer treatments.The Madison West High School SMART (Students Modeling A Research Topic) Team is Modeling RecA bound to DNA to learn more about the process of homologous recombination. Specifically, how the RecA filament interacts with DNA. Current science dictates that three RecA molecules interact with each nucleotide triplet in a DNA strand (Chen, Z., et al (2008); PDB: 3CMX), which we will be investigating in more detail. DNA replication is necessary for life, and when breaks occur in the DNA, it needs to be repaired. RecA recognizes the DNA damage, and performs homologous recombination. Understanding this process may aid in the development of novel cancer therapies, allowing for more cost‐effective treatments.
EGCG, An Active Ingredient in Green Tea, Modulates Cell Proliferation in Human Pancreatic Cancer Cells and Rat Osteosarcoma Cells in vitro
Authors: Lai Nok Hang (Tony) , Bautista Josue Kevin , Rodriguez Abigail , Bolivar Samuel , Joseph Eugene E 
Published: November 2023

Cancer is characterized by the uncontrolled proliferation of abnormal cells. Traditional treatment of cancer include surgery, radiation, and chemotherapy. Many studies have been done to elucidate the response of cancer cells to a variety of treatments. However, not many studies have been conducted on how cancer cells respond to natural compounds that are purported to have therapeutic effects on cancer cells. (‐)‐Epigallocatechin‐3‐gallate (EGCG) is a powerful ingredient of green tea which has been shown to induce apoptosis in tumor cells, prevent angiogenesis and modulate the invasion and migration of cancer cells. Pancreatic and bone cancer cells are among some of the most malignant forms of cancer. In this study, it was hypothesized that EGCG inhibits cell proliferation of both human pancreatic (Panc‐1) and rat osteosarcoma (UMR 106‐01 BSP) cell cultures, inhibits the expression of CXCR‐4, 14‐3‐3σ, and VEGFA proteins in pancreatic cancer cell cultures, and regulates mineralization in UMR 106‐01 rat sarcoma cell cultures. Panc‐1 and UMR106‐01 cell cultures were routinely passaged, treated with different concentrations of EGCG (0, 20, 40, 60, 80 and 100μM) in 6 well plates, then incubated for 48 hours at 37 degrees Celsius under humidification and 5% carbon dioxide infusion. At the end of the experiment, cells were counted using cell flow cytometry. Designated Panc‐1 cell cultures were treated with different concentrations of EGCG. After routine electrophoresis of control and experimental samples, Western blotting technique was employed to identify the expression of Panc‐1 proteins CXCR‐4, 14‐3‐3σ, and VEGFA. F test ANOVA was used to compare variances and all values in the results were expressed as means ±SD. The cell proliferation results from the Panc‐1 and UMR 106‐01 cell lines showed a decreasing trend in cell proliferation in a concentration‐dependent manner. At lower concentrations there was an increase in proliferation whereas at higher concentrations (>80uM) there was a decrease in proliferation. The results of the Western blotting technique showed that the expression of Panc‐1 proteins CXCR‐4, 14‐3‐3σ, and VEGFA are inhibited in a concentration‐dependent manner when treated with EGCG. The proliferation results suggest a dual effect of EGCG on cell proliferation both in Panc‐1 and UMR 106‐01 cancer cell cultures. EGCG may provide a therapeutic effect on both cancer cells lines at higher concentrations. The results from the Western blotting suggest that EGCG exerts its inhibitory effects on Panc‐1 cell proliferation through the regulation of the expression of the Panc‐1 proteins CXCR‐4, 14‐3‐3σ, and VEGFA.Support or Funding InformationThis research was supported by the innovations fund of the Biology Department of La Sierra University in Riverside, CA.
Gupan extracts increased the expression level of genes related to growth hormone
Authors: Kang Myung‐Hwa , Moon Hae‐Sung 
Published: November 2023

Growth hormone (GH) is the main regulator of longitudinal growth before puberty, and treatment with recombinant human GH (rec‐hGH) can increase muscle strength. Most physiological actions of GH are mediated by changes in gene transcription. GH is a 191‐amino acid pituitary protein that stimulates the hepatic production and release of insulin‐lilke growth factor‐1 (IGF‐1) into the systemic circulation. GH deficiency may occur as an isolated disorder or as part of multiple hormone deficiencies. The standard treatment of GH deficiency in adults and children is replacement with rec‐hGH. Current rec‐hGH therapy requires daily subcutaneous injections that may decrease compliance. But, these treatment evoked side effects, such as injection site reactions, edema, and arthralgia.In this study, we tested the exterio part of back (termed: Gupan) extracted from Geoclemys reevesii Gray. C2C12 murine skeletal muscle cells were cultured in DMEM supplemented with 10% FBS, 25 mM Hepes until they reached 80% confluence. Then, differentiation medium was added DMEM (5% horse serum), and the cells were allowed to differentiate into myotubes during 4 days. After 16 h in serum‐free medium, C2C12 myotubes were stimulated with Gupan (15 ug/ml) and rec‐hGH (500 ng/ml) for 6 h. RNA was obtained from each sample using Total RNA purification kit and cDNA was prepared form 250 ng of RNA using the high‐capacity cDNA reverse transcription kit. qRT‐PCR was conducted with FastStart Essential DNA Green Master (Roche). qRT‐PCT was used for confirmation of gene expression changes induced by rec‐hGH. Nine genes were confirmed as being differentially expressed after rec‐hGH induction. Among 9 genes, 3 genes (CISH, npy1r,ugdh) were highly expressed more than rec‐hGH. 2 genes (BTC, Bcl6) were also highly expressed more than no treatment. The other 2 genes (SOCS2, tgfbr3) was a little increased in the expression. However, BMP4 and RGS2 were not increased. Although we do not test many experiments, this study shows modifications in the gene expression profile of C2C12 cell line treated with Gupan in vitro. Further investigations need to be study with potential implications in the action of GH.