bluuebunny/crossref_metadata_2025_split huggingface dataset:
Co‐localization of nodal in hypoxic regions of tumours as seen using confocal microscopy and stereoscopic 3D reconstruction methods
Authors: Stock Tamara , Wilson Timothy D , Postovit Lynne‐Marie 
Published: June 2021

Metastatic breast cancer is associated with a poor clinical prognosis. Understanding the factors that promote metastasis is required so that new treatments can be derived. Studies have shown that low oxygen levels (hypoxia) characterize the microenvironment of many tumours and that tumour hypoxia is correlated with increased metastatic potential. Nodal, an embryonic growth factor, is associated with tumour progression in breast cancer. Previous work indicates that Nodal is up‐regulated in response to hypoxia. The aim of this study was to investigate the co‐localization of Nodal with areas of hypoxia within three‐dimensional (3D) tumour spheroids. A poorly metastatic, well differentiated, human breast cancer cell line (MCF‐7) was grown as mammospheres and fixed using paraformaldahyde. Nuclei, hypoxia, and Nodal were simultaneously detected using immunofluorescence. Stained mammospheres were whole mounted for optical sectioning using a confocal microscope. Images were obtained at a slice thickness of 1‐2µm. Images were compiled and rendered into a 3D image using digital segmentation software. The co‐localization of Nodal and areas of hypoxia within the mammospheres were observed. Early results indicate that there may be pockets of hypoxic regions within the mammospheres and it is anticipated that Nodal will be up‐regulated in these areas. Research support: Canadian Institute of Health Research.Grant Funding SourceInternal
Differential Distribution of Podocan Isoforms
Authors: Trubin Paul Albert , Klotman Paul , Hyink Deborah 
Published: June 2021

Podocan is a relatively novel member of the small leucine‐rich repeat family of proteins. First isolated from podocytes of HIV‐1‐infected kidney tissue, this protein seems to be involved in a number of biological pathways. Initial experiments in podocan‐knockout mice revealed increases in bone density and in vascular smooth muscle cell proliferation in response to injury when compared to the phenotypes of genetically unaltered mice. In addition, protein labeling of human kidney biopsies and vascular lesions has shown both intra and extracellular distribution of podocan. To determine if multiple isoforms of the protein exist, we began a bioinformatics study to align all reported isoforms of human podocan. We found that the previously reported 613 amino acid isoform had a strong N‐terminal signal peptide, as expected for a secreted protein. However, three other isoforms had additional amino acids in front of the signal peptide. We hypothesized that additional sequences upstream of the signal peptide would lead to retention of podocan in the cytoplasm. To test this hypothesis, we fused EGFP to the N‐ or C‐ terminus of secreted podocan. These constructs were subsequently transfected into renal 293T cells. We examined the cells by fluorescent microscopy and western blotting 48 hours later. As predicted, N‐terminal GFP‐podocan prevented efficient secretion of podocan. Our studies have led us to posit that podocan is involved in differential signaling, and this area will be the focus of future investigation. We have generated 3 C‐terminal His‐tagged podocan isoforms that have differing lengths of N‐terminal sequences upstream of the signal peptide and will use these to confirm our GFP‐fusion findings. We have also developed a series of isoform‐specific primers to help us determine whether these isoforms are differentially expressed in normal or diseased human tissue.Grant Funding SourceAmerican Association of Anatomists
Ethanol and the ubiquitin‐proteasome pathway of alveolar macrophages
Authors: D'Souza Alain Joshua , Shellito Judd E , Rudner Xiaowen L , Desai Shyamal , Ruan San Bao 
Published: June 2021

The ubiquitin‐proteasome pathway is critical in the inflammatory host response to bacterial infections. Alcohol exposure predisposes to infection, particularly pneumonia caused by Klebsiella pneumoniae (Kp). We tested the effect of alcohol on the ubiquitin‐proteasome pathway using exposure to Klebsiella pneumoniae as our model. Murine alveolar macrophage cell line (MH‐S), macrophage/monocyte cell line (RAW264.7) or primary alveolar macrophages from Balb/c mice were exposed to different alcohol concentrations and challenged with 2x106 colony forming units of heat‐killed Klebsiella. At various time points cell lysates were analyzed for 20S proteasome activity by measuring the release of proteasome‐cleaved fluorigenic substrate. We observed that 20S proteasome activity was decreased, in a dose‐dependent manner, by as much as 61% in the groups exposed to ethanol, as compared to the control group (p<0.05). Cell lysates were also assayed, by Western analysis, for levels of total intracellular free protein ubiquitin and of polyubiquitinated proteins. We observed that alcohol alters levels of intracellular polyubiquitinated proteins but does not affect levels of intracellular free ubiquitin. We conclude that pulmonary infection caused by Klebsiella may be enhanced by alcohol exposure through inhibition of the ubiquitin‐proteasome pathway of alveolar macrophages.Supported by AA09803, HL076100.
Exploring leucine‐rich repeat proteins as molecular scaffolds: design of an oxidation‐resistant ribonuclease inhibitor
Authors: Rowland Elizabeth Fay , Krawisz Anna , Dickson Kimberly Anne 
Published: June 2021

Engineered proteins with specific binding activities have evolved as a powerful technology. Repeat proteins, which exhibit structurally conserved modular frameworks and extensive surface areas, are alluring molecular scaffolds; some have yielded molecules with the specificity and robustness of antibodies. Our long‐term goal is to design novel binding surfaces on the human ribonuclease inhibitor (RI), a horseshoe‐shaped leucine‐rich repeat (LRR) protein that binds to pancreatic‐type ribonucleases with extraordinary affinity (KD ? fM). Exploration of RI as a protein scaffold has been limited by its oxidation sensitivity.We have constructed oxidation‐resistant variants of RI in which some or all of its 32 cysteine residues have been replaced. Six variants of RI have been produced in which groups of 5‐7 cysteines (at conserved positions within the LRR motif) have been mutated. Preliminary results indicate that the RI variants maintain affinity for ribonucleases, but possess decreased oxidation sensitivity. These data contribute to our understanding of the structural stability of LRR proteins and suggest that RI and other LRR proteins are practical scaffolds for the design of novel binding surfaces. This work was supported by a Monticello College Foundation Research Grant to E.F.R., and start‐up funds (Macalester College and Lawrence University) to K.A.D.
Possible Biochemical Mechanism of Action of Petroleum‐Ether Extract of Calotropis procera leaves (Asclepiadaceae) in Gravid Pregnant Sprague Dawley Rats
Authors: James Ayorinde Babatunde , Magbagbeola Olubunmi Adeola , Saibu Gbemisola Morounke , Oloyo Ahmed 
Published: June 2021

Calotropis procera (Asclepiadaceae) is one of the traditionally used antifertility plants in Nigeria. Previous studies have shown that this antifertility plant has abortifacient property but none of them has reported its possible biochemical mechanism of action.Organ bath experiments using cumulative doses of the extract on rat uterine rings in Dejalon's solution aerated with 95% O2 + 5% CO2 produced a non significant (P>0.05) increase in tension (% response). An EC50 value of 0.1059 mg/ml was obtained and the intrinsic activity was found to be above 1 in comparison with oxytocin.Blocking muscarinic receptors with 35µg/ml atropine caused the dose‐response curve to shift to the right with a non‐significant (P>0.05) decrease in maximal tension (% Response). However, the ?2‐adrenergic blocker (1µM propanolol) caused a significant (P<0.05) shift in the dose‐response curve.The extract also exhibited its effect on cAMP modulation of β2‐adrenergic receptors by causing a significant (P<0.05) decrease in cAMP level after treatment of cultured uterine cells with the extract.In conclusion, this study suggests that pet‐ether extract of Calotropis procera (Asclepiadaceae) exerts its abortifacient effect by inducing myometrial contractions of the uterus by binding to β2‐adrenergic receptors thereby causing a decrease in the level of cAMP. cAMP reduction may reduce the activation of protein kinase A (PKA) preventing PKA from inactivating myosin light chain kinase (MLCK).
Medicinal use of vinegar should be accompanied by a diet rich in potassium alkali salts to avoid possibility of low grade metabolic acidosis
Authors: Appel Christy L , Johnston Carol S 
Published: June 2021

Although daily ingestion of vinegar may help manage blood glucose, vinegar can decrease urine pH and possibly exacerbate low grade metabolic acidosis (LGMA). We examined if daily ingestion of vinegar or control would decrease urine pH in a randomized, crossover trial. Because change in urine pH may be buffered by dietary intake of potassium (K) alkali salts, we also determined the impact of K status on urine pH during the trial. Healthy subjects (3 M, 7 F, age 27.7±3.1 y [mean±SEM], BMI 24.9±2.0 kg/m2) ingested 2 tablespoons of vinegar (VIN) or cranberry juice (CON) twice daily for 3 d with a 2 wk washout period. While no pH change was noted with the CON treatment, mean 24 h urine pH after 3 d with the VIN treatment was significantly more acidic if K status was low versus adequate (pH 5.62±0.10 versus 6.24±0.11, P=0.015). Low K status was defined as baseline 24 h urine K <50 mmol/L (n=3). Thus, it appears that high intake of K‐alkali salts enhances base‐forming capacity to buffer the acid load associated with vinegar. Persons using vinegar medicinally may benefit from consuming vinegar with K‐alkali rich plant foods to avoid LGMA. This research was supported by the ASU Nutrition Research Fund.Grant Funding SourceASU Nutrition Research Fund
Effects of Moderate Zinc Deficiency and Phytase Supplementation on Lumbar Vertebrae in Exercising Rats
Authors: Catrambone Daniel E , Antczak Amanda J , Hendrickson Nathan R , Regalia Ryan S , Young Andrew J , Scrimgeour Angus G 
Published: June 2021

Dietary zinc (Zn) is important for bone remodeling. Phytase (P) increases the bioavailability of Zn and other micronutrients. How modeling adaptations of the lumbar spine may be affected by a P supplemented low Zn diet has not been investigated.PURPOSEDetermine the effects of a low Zn diet and supplemental P on the 4th lumbar vertebra (L‐4) in exercising rats.METHODS30 Male Sprague‐Dawley rats were housed in cages fitted with voluntary running wheels and fed either a 5ppm Zn (ZnLo), 5ppm Zn + 1,500 FTU/kg P (ZnLo+P) or a 30ppm Zn (ZnAd) diet for 9 wks, (n=10 per group). In vivo peripheral quantitative computed tomography scans were performed at 40, 50 and 60% of the total L‐4 length. Cortical content (CNT) (mg), area (A) (mm2) and bone mineral density (BMD) (mg/cm3) were measured. Plasma concentrations (PC) (µg/ml) of Zn were determined. A one‐way ANOVA was used to compare the data.RESULTSZnLo+P exhibited increased CNT (11.6 ± 1.5 vs. 9.9 ± 0.7, p=0.006), A (12.0 ± 1.4 vs. 10.5 ± 0.7, p=0.007) and BMD (959.5 ± 15.9 vs. 938.8 ± 7.9 p=0.012) compared to ZnLo and increased BMD (959.5 ± 15.9 vs. 942.5 ± 18.9, p=0.043) compared to ZnAd. ZnAd exhibited increased PC of Zn (1.2 ± 0.2 vs. 0.8 ± 0.1 and 0.5 ± 0.2, p<0.001) compared to ZnLo+P and ZnLo respectively. ZnLo+P exhibited increased PC of Zn (0.8 ± 0.1 vs. 0.5 ± 0.2, p=0.01) compared to ZnLo.CONCLUSIONP supplementation increased PC of Zn and increased mid‐vertebral cortical CNT, A and BMD in a ZnLo fed sample. The greatest L‐4 cortical modeling was observed in rats fed a ZnLo+P diet.The study was funded by the U.S. Army MRMC
Role of dietary glutamic acid in gut‐immune system: impaired immune responses in mice fed glutamic acid‐free diet
Authors: Yoneda Junya , Kawamata Yasuko , Chin Keigi , Torii Kunio , Sakai Ryosei 
Published: June 2021

Although sufficient intake of protein and amino acids is essential for normal immune functions, it has not been elucidated well whether dietary glutamic acid (Glu), the most abundant amino acids in food proteins, plays any roles in the maintenance of immune functions or not. Therefore, this study aimed to assess the effect of removal of Glu from the diet on mucosal and systemic immune responses in immunized mice.Female BALB/c mice were divided into two dietary groups, and were given amino acids defined diet (control) or Glu‐free diet during one‐week acclimation period and successive immunization period. The mice were orally immunized with 2mg of keyhole limpet hemocyanin (KLH) and 10μg of cholera toxin (CT) on day 0, 10 and 20 or systemically immunized (i.p.) with 100μg of KLH and 1μg of CT on day 0 and 14 (n=5 for each). One week after the last immunization, plasma samples were collected to measure KLH‐specific antibody titer by ELISA. Lamina propria lymphocytes (LPL) were harvested to assess their IgA‐secretion in vitro (ELISPOT assay).Feeding of Glu‐free diets reduced the elevation of antigen‐specific antibody titer in plasma in orally immunized mice, which was accompanied with reduced IgA‐secreting activity in mucosa. Besides, there was no effect in the responses against systemic immunization. These results suggest nutritional roles of Glu in the maintenance of normal mucosal immune response.
Canola oil is protective against macrophage infiltration into subcutaneous adipose tissue of swine
Authors: Braucher Doug R , Gabler Nicholas K , Walker‐Daniels Jennifer , Davis Jeremy E , Spurlock Michael E 
Published: June 2021

Omega 3 fatty acids (FA) possess anti‐inflammatory properties and attenuate inflammatory events linked to obesity, including macrophage infiltration into adipose tissue. The anti‐inflammatory properties of dietary n‐3 fatty acids (FA) have been primarily attributed to dietary sources of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA). However, the potential benefit of dietary α‐linolenic acid (αLA) to alleviate inflammation has not been determined. We hypothesized that EPA and DHA production from dietary αLA metabolism would be sufficient to attenuate inflammation in pigs fed a high fat diet. Pigs were fed a diet containing either 25.4% canola oil, 28% lard, or 0.5% canola and 0.5% lard (control) for 11 weeks. Gas chromatography detected EPA and docosapentaenoic acid (DPA) in the liver tissue of the canola and control diet fed pigs. EPA was also in the subcutaneous (SQ) adipose tissue of pigs fed canola oil. DHA was not detected in any tissue. Macrophage infiltration was assessed based on expression of the porcine macrophage specific gene swine workshop cluster 3 (SWC3) as determined by quantitative PCR. SWC3 transcript abundance in SQ adipose tissue was lower in pigs fed the canola oil vs. those fed the lard (p=0.04) or control (p=0.03) diets. These data suggest that a diet rich in αLA creates a tissue specific fatty acid profile that protects against the infiltration of macrophages into SQ adipose tissue.Grant Funding SourceIowa State University
Splenic dendritic cell populations in vitamin A deficient C57BL/6J mice
Authors: Duriancik David Michael , Hoag Kathleen Ann 
Published: June 2021

Vitamin A‐deficient individuals have altered adaptive immunity, showing depressed antibody‐mediated [T helper (Th)2] responses and enhanced cell‐mediated (Th1) responses. In the mouse, myeloid dendritic cells (DCs) and lymphoid DCs are responsible for stimulating Th2 and Th1 responses, respectively. The objective of this research project was to compare the percentage and absolute numbers of DC populations in vitamin A‐sufficient and vitamin A‐deficient uninfected laboratory C57BL/6J mice. The hypothesis was that vitamin A‐deficient animals would have decreased myeloid DCs and increased neutrophils and precursor DCs compared to vitamin A‐sufficient animals. We used dietary restriction to deplete the animals of vitamin A and multicolor flow cytometry to detect and quantify the numbers of immune cell populations from individual mouse spleens. In accordance with our hypothesis, the myeloid DCs are decreased and the neutrophils and precursor DCs are increased in the vitamin A‐deficient animals compared to the vitamin A‐sufficient animals. Therefore, the observed decrease in myeloid DCs in vitamin A‐deficient animals may be the basis for depressed Th2 function in vitamin A‐deficient animals.Grant Funding SourceDepartment of Food Science and Human Nutrition, Michigan State University
n‐3 Polyunsaturated Fatty Acids Suppress T‐Cell Mitochondrial Translocation to the Immunological Synapse
Authors: Yog Rajeshwari , Barhoumi Rola , McMurray David N. , Chapkin Robert S. 
Published: June 2021

T helper (Th) cell activation is necessary for the adaptive immune response. Formation of the immunological synapse (IS) between Th cells and antigen‐presenting cells is the first step in Th cell activation. In vitro studies indicate that formation of the IS induces cytoskeleton‐dependent mitochondrial redistribution to the immediate vicinity of the IS. This redistribution of mitochondria to the IS in T cells is necessary to maintain Ca2+ influx across the plasma membrane and Ca2+‐dependent Th cell activation. In an earlier study, we demonstrated that n‐3 polyunsaturated fatty acids (PUFA) suppress the localization and activation of signaling proteins at the IS. Therefore, we hypothesized that n‐3 PUFA suppress CD4+ T‐cell mitochondrial translocation during the early stages of IS formation. CD4+ cells derived from fat‐1 mice, a transgenic model that synthesizes n‐3 PUFA from n‐6 PUFA, were co‐cultured with anti‐CD3‐expressing hybridoma cells (145‐2C11) for 15 min at 37°C, and mitochondrial relocation to the IS was assessed by confocal microscopy. T‐cells from fat‐1 mice significantly reduced the percentage of cells with mitochondria which translocated to the IS; fat‐1 (34%) vs. wild type control (85%); P=0.008. These results demonstrate that n‐3 PUFA limit mitochondrial translocation to the IS, a critical early step for CD4+ T‐cell activation.Supported in part by NIH grant DK07107, CA59034 and P30ES09106Grant Funding SourceNIH DK71707 and CA59034
Incidence of Obesity and Associated Health Risks in the District of Columbia Public Schools
Authors: Lockwood Susan D , Harland Barbara F , Steinert Donald , Tavakol Karman 
Published: June 2021

A Youth Risk Behavior Study (YRBS) was conducted by the Center for Disease Control and Prevention in collaboration with the District of Columbia (DC) Public Schools and the DC Department of Health. Twenty‐seven percent of DC high school students were either overweight or obese with BMI's over 25. Eighty percent did not meet recommended levels of physical activity. These values are higher than corresponding values for the national average. Physical activity is important for both short‐ and long‐term maintenance of normal body weight. Physical activity appears to have an independent effect on health‐related outcomes in addition to weight control. These negative health conditions may substantially increase the risk of morbidity from hypertention, dyslipidemia, type 2 diabetes, coronary artery disease, stroke, gallbladder disease, osteoarthritis, sleep apnea, asthma, respiratory problems and cancer. Overweight youngsters are permitting TV, video games, and indiscriminate eating to take the place of regular, controlled regimes including vigorous physical activity accompanied with intelligent food selection.A proposal has been developed for the study of obesity among high school children grades 9 through 12 in the DC public schools, via a survey of 3 schools from poorer neighborhoods, and 3 schools from wealthier areas (1200 subjects). Heights, weights, BMI's and blood pressure will be recorded. Faculty and students from the University of the District of Columbia and Howard University will officiate in an attempt to interrupt these negative behavior patterns.Funded by Research Infrastructure in Minority Institutions (RIMI).
Evolutionary conservation of Tino/Quaking interaction suggests its impact to cell polarity control in ontogenesis and oncogenesis
Authors: Lulli Matteo , Donnini Martino , Lapucci Andrea , Quaglierini Paolo , Witort Ewa , Papucci Laura , Specogna Rebecca , Morello Dominique , Capaccioli Sergio 
Published: June 2021

Loss of cell polarity is very common in invasive and metastatic tumours, but also in early stages of cancerogenesis. In C. elegans the conserved Par genes control several proteins transducing polarity cues in the embryo. One of these transducers is ceMex‐3, a translational regulator of cell fate during development. Its human ortholog is Tino, firstly identified in our lab as a post‐transcriptional regulator. The high conservation between ceMex‐3 and Tino suggests that also Tino could have an impact on mammalian embryogenesis, cell polarity and cancer. Par‐4 is a ceMex‐3 translational regulator, its human ortholog Lkb1 is responsible, if mutated, for the Peutz‐Jeghers cancer syndrome. Another liaison dangereuse co‐involves ceMex‐3 with Gld‐1 gene, whose mammal ortholog is Quaking (Qki). Both ceMex‐3 and Gld‐1 co‐regulate common targets and if mutated induce onset of teratomas during embryogenesis.Here we show that Qki contributes to the negative regulation of Tino, confirming our hypothesis of evolutive conservation of Tino pathway. Furthermore, identification of Tino's target mRNAs and their consensus reveals that 50% of these mRNAs contains also Qki consensus, underlying the possibility that Qki and Tino co‐regulate common targets.
Role of Phox2B in neuroblastoma development
Authors: Alam Goleeta Nazaneen 
Published: June 2021

Neuroblastoma (NB) a common childhood malignant tumor of the sympathetic nervous system presents as a heterogeneous group of tumors ranging from tumors comprising of undifferentiated neuroblasts to those largely consisting of fully differentiated neurons. We have used MYCN transgenic mice‐ mice with targeted expression of the oncogene MYCN in neural crest cells‐ as a model system to investigate the cellular basis for NB development. These mice develop overt neuroblastomas preceded by a pre‐neoplastic stage characterized by development of hyperplastic lesions in the post‐natal sympathetic ganglia. Immunostaining analysis of these lesions revealed extensive proliferation and absence of staining with sympathetic differentiation markers, indicating their undifferentiated state. A majority of the hyperplastic cells as well as majority of cells in all primary tumors examined showed strong staining for Phox2B, a marker of sympathetic neural progenitors. Recently, we observed that retinoic‐acid induced differentiation of human NB cell lines is accompanied by a marked down‐regulation of Phox2B suggesting that Phox2B may be important for maintenance of the neuroblasts characterizing NB tumors. Also, Phox2B down‐regulation in the human NB cell line SK‐N‐AS results in decreased survival of cells. Given together, these data suggest that Phox2B may function as a lineage‐survival oncogene in NB development.NCI