bluuebunny/crossref_metadata_2025_split huggingface dataset:
Serum and glucocorticoid inducible kinase regulates hERG channels through inhibition of Nedd4–2 and activation of Rab11 recycling endosomes
Authors: Lamothe Shawn Michael , Zhang Shetuan 
Published: June 2021

The human ether‐a‐go‐go‐related gene (hERG) encodes the rapidly activating delayed rectifier potassium channel (IKr). Loss of function of hERG channels due to mutations or drug‐induced blockage can cause long QT syndrome, leading to ventricular arrhythmias or sudden death. Physical or emotional stress is known to affect ion channel activity. Stress hormones such as cortisol regulate the expression of various genes including the serum‐ and glucocorticoid‐inducible kinase (SGK). Using patch clamp, Western blot, co‐immunoprecipitation, and immunocytochemistry methods, we demonstrate that SGK isoforms SGK1 and SGK3 increased hERG current (IHERG) and the expression level of mature hERG proteins. We recently showed that mature hERG channels are degraded by ubiquitin ligase Nedd4–2 via enhanced ubiquitination. We observed that SGK1 and SGK3 overexpression enhanced Nedd4–2 phosphorylation which is known to inhibit Nedd4–2 activity. However, removal of the Nedd4–2 effect on hERG channels by disrupting Nedd4–2 target motif in hERG reduced but did not eliminate the SGK‐induced increase in hERG expression. Additional disruption of Rab11 proteins led to a complete elimination of SGK‐mediated hERG increase. We conclude that SGK isoforms 1 and 3 enhance hERG stability by inhibiting Nedd4–2 ubiquitin ligase and activating Rab11 recycling endosomes.Supported by: Canadian Institutes of Health Research (CIHR)
Inflammatory cytokines in human milk are inter‐correlated and may be related to infant growth characteristics and maternal weight status
Authors: Young Bridget E. , Morrow Ardythe L. , Davidson Barbara S. , Geraghty Sheela R. , Krebs Nancy F. 
Published: June 2021

This ongoing study aims to longitudinally characterize inflammatory cytokines in human milk (HM) and investigate associations with maternal weight and infant growth characteristics. Morning HM samples from 9 normal weight (NW) and 9 overweight (OW) mothers (BMI range: 21.0 – 30.0 kg/m2) and anthropometry measures of their healthy term infants were obtained at 2‐weeks (T1: 2.1 ± 0.4 wks) and 4‐months (T2: 16.7 ± 1.2 wks) post‐partum. HM % fat was measured by creamatocrit; IL‐6, IL‐8, and TNF‐α concentrations by multiplex. The inflammatory load of HM was calculated as the standardized sum of cytokine concentrations. Infant length for age, weight for age, and weight for length Z‐scores (LAZ, WAZ, WLZ) did not significantly change from T1 to T2. IL‐8, TNF‐α, and inflammatory load at T1 were positively correlated with those at T2 (p < 0.01, R2≥ 0.39). IL‐6 and TNF‐α decreased from T1 to T2 (p < 0.05). IL‐6, TNF‐α, and % fat did not differ between NW and OW mothers. HM IL‐8 and inflammatory load decreased from T1 to T2 in NW (p = 0.03) but tended to increase in OW mothers (p = 0.07). By T2, all 3 cytokines were positively inter‐correlated (p < 0.02, R2≥ 0.33). Infant growth results were inconclusive: HM inflammatory load at T1 inversely correlated with ΔWAZ from T1 to T2 (p = 0.04, R2= 0.26) but not ΔLAZ (p = 0.76) or ΔWLZ (p = 0.05). Maternal BMI status may impact inflammatory cytokines in early HM.Grant Funding Source: NIH T32DK007658–21, 5K24DK083772‐08, P01HD13021
Effects of fructose overfeeding on intrahepatic lipid accumulation and hepatic insulin sensitivity in healthy humans
Authors: Lecoultre Virgile , Egli Leonie , Boss Andreas , Carrel Guillaume , Theytaz Fanny , Zwygart Karin , Kreis Roland , Schneiter Philippe , Boesch Chris , Tappy Luc 
Published: June 2021

Added sugars may contribute to the development of insulin resistance in metabolic diseases. It is further suspected that this is related to fructose‐induced intrahepatic fat deposition. How it relates to daily fructose intake, and what the relative roles of fructose, glucose and excess energy are, remains however controversial. We therefore evaluated the effects of short term overfeeding with various amounts of fructose, glucose, or saturated fat on hepatic insulin sensitivity (HIS) and lipid content (IHCL) in healthy male subjects. Data collected in several studies were pooled. Fasting hepatic glucose production (HGP), HIS and IHCL were measured after both 6–7 days on a weight‐maintenance diet (control, C; n=53) and 6–7 days of overfeeding with 1.5 (F 1.5, n=7), 3 (F3, n=17), or 4g fructose/kg/day (F4, n=10), with 3g glucose/kg/day (G3, n=11), or with 30% excess energy as saturated fat (fat 30%, n=8).F3, F4, G3 and fat 30% all significantly increased IHCL as compared to C (all p<0.05). F4 and G3 significantly increased HGP, (both p<0.05), and F3 and F4 significantly decreased HIS (both p<0.01). In contrast, there was no significant effect of fat 30% on HGP or HISI.Overfeeding with large amounts of fructose or glucose impairs fasting hepatic glucose metabolism and increases hepatic fat content within 6–7 days. This indicates short‐term regulation of hepatic glucose metabolism by simple carbohydrates.
Macrophage apolipoprotein E and proliferation of breast cancer cells MCF‐7: Role of LXR
Authors: Bard Jean‐Marie , El Roz Ali , Valin Sabine , Huvelin Jean‐Michel , Nazih Hassan 
Published: June 2021

ObjectiveApolipoprotein (apo) E is expressed by macrophages under control of LXR, a transcription factor. LXR agonists may inhibit cancer cell proliferation in vitro. We evaluate the effect of apo E, secreted by macrophages under control of LXR on MCF‐7 cell proliferation, a model of breast cancer cells.MethodsMacrophages derived from THP‐1 monocytes were incubated with LXR agonists [22(R)‐hydroxycholesterol and TO901317] before (THP‐1 + agonists) and after (THP‐1 + siRNA apo E) deprivation of apo E by siRNA. MCF‐7 cells were incubated with media from each macrophage model, as well as exogenous apo E (20 μg/mL). Cell viability was tested by MTT method and apoptosis gene expression (BAX, Bcl‐2) was analysed by qPCR.ResultsApo E, as determined by ELISA and qPCR is increased in “THP‐1 + agonsits” medium and decreased in “THP‐1 + siRNA apo E” medium. MCF‐7 cell viability is decreased by 18 % and 40% after 24h and 48h incubation with exogenous apo E. Medium from “THP‐1 + agonists” decreases MCF‐7 cell viability by 30% after 24h incubation, while medium from “THP‐1 + siRNA apo E” has no effect. The pro‐apoptotic gene BAX is overexpressed in MCF‐7 cells treated with medium from “THP‐1 + agonists” and underexpressed when treated with medium from “THP‐1 + siRNA apo E”.ConclusionApo E from macrophages stimulated by LXR agonists may inhibit cell proliferation and induce apoptosis in MCF‐7, a model of breast cancer cells.
Vitamin A‐mediated inhibition of cell cycle progression proteins and cell growth requires protein‐bound retinol in a prostate cancer cell line
Authors: Colley Danielle L , Castonguay Thomas W 
Published: June 2021

Stimulated by retinoic acid homolog 6 (STRA6) is a transmembrane receptor specific for retinol‐binding protein 4 (RBP4) that mediates uptake of retinol bound to RBP4 (HoloRBP4). Retinoic acid derivatives of retinol have well‐documented anti‐proliferative actions that are mediated by retinoic acid and retinoid X receptors (RAR and RXR, respectively). Thus, it has long been assumed that the effects of circulating retinol on the regulation of apoptosis are dependent on cellular uptake of retinol and activation to retinoic acid. An additional role of STRA6 has recently been identified in vivo and in cultured hepatocytes and renal proximal tubule cells. Upon binding to holoRBP4, STRA6 has been shown to activate a cascade that led to STAT5‐mediated gene regulation. We have detected and induced STRA6 with all‐trans‐retinoic acid (RA) in PC‐3 prostate cancer cells. In this study, we found that PC‐3 cells were growth inhibited following treatment with holoRBP4 whereas 1 μmol/L RA treatment had no effect. Furthermore, we observed that holoRBP4 treatment reduced mRNA and protein abundance of cell cycle progression proteins (CD1, CDK2, and CDK4), which have been identified as a downstream target of STAT5. Collectively, these results indicate that anti‐proliferative actions of retinol may be produced via a signal transduction mechanism, which appear to occur independently of RA‐mediated gene regulation.Grant Funding Source: National Institutes of Health
Development of respiratory tract infection could be modified by the interactions between maternal diet during pregnancy and offspring's CD14 (rs#2569190) and VDR (rs#7975232) polymorphisms
Authors: Hong Seo Ah , Kwon Sung Ok , Kim Kyung Won , Shin Youn Ho , Ahn Kang Mo , Yu Jinho , Kwon Ja‐Young , Choi Suk‐Joo , Lee Kyung‐Ju , Won Hye‐Sung , Oh Se‐young , Hong Soo‐Jong 
Published: June 2021

Maternal diet during pregnancy can potentially influence disease susceptibility of offspring in addition to maternal atopy and genetic background. The objective was to investigate the association of prenatal antioxidant intakes and the genotype with the risk of respiratory tract infection (RTI) in 12‐month‐old infants. As part of a birth cohort study, the COhort for Childhood Origin of Asthma and Allergic Diseases (COCOA) (n=528), we assessed maternal diet during pregnancy by FFQ and genotyping of 4 genes (CD14, VDR, NRF2 and GSTP1) by the Taqman method. Maternal intakes of vit A and C, Fe, folic acid, and vegetable were negatively associated with the risk of RTI, particularly in the infants with atopic mothers (p<0.05). In combined analyses with genotype, we found significant interactions between the genetic variation in CD14 and vit A and C and fruit and between the genetic variation in VDR and fruit and vegetable regarding RTI (p<0.05). These findings suggest that the role of prenatal intakes of antioxidant nutrients related to lower risk of RTI may be modified by CD14 and VDR polymorphisms in infants.This study was supported by a fund (2008‐E33030–00, 2009‐ E33033–00, 2011‐E33021–00) from the Research of Korea Centers for Disease Control and Prevention.Grant Funding Source: Nutrition (ASN)
Changes in Leukocyte Populations Among Children Receiving a Follow‐on Cow's Milk‐Based Formula Containing Beta‐Glucan
Authors: Sarvetnick Nora , Murphy Kevin , Pauley‐Hunter Rosemary , Scalabrin Deolinda 
Published: June 2021

Backgroundβ‐Glucan can enhance immune system responses to foreign agents, but mechanisms of action remain unknown.ObjectiveWe examined changes in immune cell populations and activation markers produced by the addition of β‐Glucan to a children's follow‐on formula.MethodsThis was a double blinded, randomized, controlled, parallel‐designed, prospective study. Data is reported for participants who completed the study. Participants (1–4 years of age) received either a marketed, follow‐on formula (n=8), formula supplemented with 42 mg of β‐Glucan per serving (n=10), or formula with 6 × 109 colony forming units of LGG per serving (n=9), twice a day for a 3 week period. Flow cytometry was used to immunophenotype peripheral blood leukocytes. Thirty serum cytokines were measured via multiplex analysis. Changes from baseline were analyzed by Kruksal‐Wallis. As this was a pilot study, differences with p‐values < 0.10 were considered of interest.ResultsParticipants who received β‐Glucan had decreased production of stem cell factor and decreased neutrophil proportions. They also had an increase in the ratio of immature‐to‐mature B cells. Finally, they had reduced CD4 T cell activation and an increase in specific subsets of CD8 T cells.Conclusionβ‐Glucan promotes a complex pattern of activation and inhibition of distinct T and B cell subsets. To our knowledge, this is the first study to examine the effects of β‐Glucan on immune effects in healthy, young children.
Histone deacetylase (HDAC) inhibition prevents and restores bacterial lipopolysaccharide (LPS)‐induced paracellular hyper‐permeability in human lung microvascular endothelial cells (HLMVEC)
Authors: Joshi Atul D , Thangjam Gagan , Barabutis Nektarios , Shaw Mary , Patel Vijay , Catravas John D 
Published: June 2021

ObjectiveLPS disrupts the barrier function of endothelial cell monolayers and increases their permeability to proteins. We have previously shown that inhibition of hsp90 protects and restores LPS‐mediated hyperpermeability in human lung microvascular endothelial cells (HLMVEC). Inhibition of histone deacetylases (HDAC) hyperacetylates hsp90 and thus suppresses its chaperone function. In this study we investigated the effects of HDAC inhibition on LPS‐mediated endothelial barrier disruption.MethodsEndothelial permeability was measured by electric cell‐substrate impedance sensing (ECIS), in HLMVEC grown to confluence on gold‐plated arrays.ResultsPre‐treatment with the HDAC inhibitors (TSA, butyrate or panobinostat) protected HLMVEC monolayers from LPS‐mediated hyperpermeability. Treatment with either TSA (2μM) or panobinostat (1μM) 3–6 hours after LPS exposure, restored endothelial barrier function. In addition, LPS‐mediated phosphorylation of MLC and hsp27 was attenuated by pre‐treatment with TSA or panobinostat, possibly preventing cytoskeleton rearrangement. TSA also disrupted hsp90‐CDC37 interaction, in the presence or absence of LPS, indicating a loss of hsp90 chaperone function.ConclusionHDAC inhibition protects and restores LPS‐mediated endothelial barrier function possibly by inhibiting hsp90 chaperone function. (Supported by HL093460 and HL101902)
Giardia duodenalis cysteine cathepsin proteases and their role in intestinal disease
Authors: Bhargava Amol , Cotton James A. , Yates Robin , Buret Andre G. 
Published: June 2021

Giardia duodenalis, an enteric protozoan parasite, induces pathophysiological effects shared with chronic gastrointestinal disorders, including barrier function disruption, via unknown mechanisms. Cysteine proteases, also identified in the Giardia genome, are implicated in disease pathophysiology of other protozoan parasites.ObjectiveTo characterize and determine a role for cysteine cathepsins in the pathogenesis of giardiasis.ResultsIn vitro cathepsin B/L activity was detected within Giardia trophozoites and in co‐culture supernatants. Giardia trophozoites increased cathepsin B activity in host enterocytes during co‐cultures; this required trophozoite adhesion to host cells. Pretreating Giardia trophozoites with Ca‐074Me (a cathepsin B specific inhibitor) prevented elevation of cathepsin activity in host cells. Inhibition of Giardia cathepsins with a cysteine protease inhibitor (E64) prevented the alteration of the mucus layer glycoprotein, MUC‐2; however, Ca‐074Me failed to prevent Giardia‐induced ZO‐1 cleavage.ConclusionGiardia trophozoites increase cathepsin activity in host cells and express cathepsins that play a role in mucus alteration. Supported by NSERC, AIHS, CCFC.
Effects of exposure do air pollution on placental structure
Authors: de Fatima Soto Sonia , Lopes Karen , Veras Mariana Matera , Heimann Joel 
Published: June 2021

ObjectiveTo evaluate the effects of exposure to air pollution during the pre‐gestational or gestational period.MethodsFemale Wistar rats (7–9/group) were exposed to filtrated (F) or polluted (P) air in a concentrator of atmospheric particles for 3 weeks. At 12 weeks, after mating, rats were divided in 4 groups exposed to F or P (FF, FP, PF, PP). At 19th day of pregnancy: fetal placenta (Pl), decidua (Dc) and fetus (F) mass; Pl and Dc longitudinal (LD), transversal (TD) diameters and thickness (T); Pl, Dc and serum ACE activity; serum Aldosterone were measured.Results (mean±SEM – p<0.05):

FF
FP
PF
PP

Mass (g)
Pl+Dc
0.63±0.010
0.61±0.009
0.57±0.0141
0.55±0.0071

Pl
0.42±0.006
0.44±0.006
0.40±0.0052
0.43±0.004

Dc
0.13±0.005
0.14±0.008
0.12±0.0051
0.12±0.0071

Diameters ratio (LD/TD)
Pl
1.08±0.02
1.24±0.043
1.11±0.02
1.17±0.02

Dc
1.07±0.02
1.16±0.03
1.10±0.03
1.25±0.043

T (mm)
Pl
4.42±0.10
4.16±0.06
3.93±0.094
4.40±0.11

Dc
3.15±0.10
2.77±0.095
2.92±0.09
2.95±0.11

ACE (nmol His‐leu/min/mL)
Pl
1.05±0.07
0.99±0.11
1.12±0.10
1.35±0.24

Dc
0.83±0.07
0.67±0.06
0.59±0.055
0.53±0.055

Serum
33.50±1.39
28.89±1.375
28.90±1.675
26.53±1.065

vs FF, FP;
vs FF, FP, PP;
vs FF, PF;
vs FF, PP;
vs FF.

Serum Aldosterone was similar among all experimental groups.ConclusionsExposure to air pollution affects placental structure indicating a possible prejudice of fetal nutrition.Supported by FAPESP and CNPq.
Novel use of a three‐dimensional in vitro environment; does flexor and extensor tendon cell behavior differ?
Authors: Stanley Rachael Louise , Carolin Amy Miriam , Agabalyan Natasha Anahid , Evans Darrell John Rhys 
Published: June 2021

Flexor tendons commonly suffer strain‐related injuries that are challenging to treat. Extensor tendons rarely suffer such damage. Investigating the differences between these tendon types may help us develop new treatments. Traditional 2‐dimensional culture of tendon cells does not appear to adequately replicate the 3D structure of tendons; therefore we investigated the differences of chick flexor and extensor tendon cell behavior in 2D and 3D environments. By creating cylindrical collagen gel structures seeded with separate cell types, we examined cell morphology and extracellular matrix (ECM) protein expression. Results demonstrated that in a 3D environment, both cell types and ECM proteins were arranged in the direction of tension; a pattern similar to tendons in vivo. Our results show that use of a 3D gel construct is an easy and reproducible method for future in vitro tendon research. To our knowledge, this is the first study to compare flexor and extensor cells in a 3D environment. Funded by The Leverhulme Trust, The Wellcome Trust and Brighton and Sussex Medical School.Grant Funding Source: N/A
Histone Acetyl Transferase (HAT) HBO1 and PHD Finger Protein 17 (PHF17/JADE1) in Epithelial Cell Regeneration
Authors: Panchenko Maria , Havasi Andrea , Haegele Joseph , Bonegio Ramon , Ichimura Takaharu , Siriwardana Nirodhini , Gall Jonathan 
Published: June 2021

HBO1 acetylates lysine residues of histones and is involved in DNA replication and gene transcription. Two isoforms of JADE1, JADE1S and JADE1L, bind HBO1 and promote acetylation of chromatinal histones. We characterized the role of JADE1‐HBO1 complexes in vitro and in vivo during epithelial cell replication. Downregulation of JADE1 by siRNA diminished the rate of DNA synthesis in cultured cells, decreased endogenous HBO1 protein expression, and prevented chromatin recruitment of replication factor Mcm7, demonstrating that JADE1 is required for cell proliferation. We used a murine model of acute kidney injury (AKI) to examine expression of HBO1‐JADE1S/L in injured and regenerating epithelial tissue. In control kidneys, JADE1S, JADE1L, and HBO1 were expressed in nuclei of proximal and distal tubular epithelial cells. Ischemia and reperfusion injury resulted in an initial decrease in JADE1S, JADE1L, and HBO1 protein levels which returned to the baseline during renal recovery. HBO1 and JADE1S recovered as cell proliferation reached its maximum while JADE1L recovered after bulk proliferation had ceased. The temporal expression of JADE1S correlated with the acetylation of histone H4 on lysine 5 and 12, but not with acetylation of histone H3 on lysine 14, demonstrating that the JADE1S‐HBO1 complex specifically marks H4 during epithelial cell proliferation. These data implicated JADE1‐HBO1 complex in AKI and suggested distinct roles for JADE1 isoforms during epithelial cells recovery.NIH RO1 DK087910, and ACS IRG‐72–001‐32 (to M. V. P.). NIH 1KO8DK090143 (to A.H.).
Prevention of ubiquitination at lysines in the N tail, R, ICL4, and NBD2 domains disrupts CFTR trafficking and immune response
Authors: Lee Seakwoo , Schiffhauer Eric , Kang Po Wei , Zeitlin Pamela 
Published: June 2021

RationalePost‐translational modification of lysine residues in CFTR is predicted to regulate CFTR trafficking and recycling. We hypothesized that disruption of ubiquitination in lysine residues will accelerate destruction of wt CFTR and increase inflammatory responses.MethodsSite‐directed mutagenesis, Immunoblot, confocal analysis, ELISA assay, etc.ResultsWe identify seven key lysines in CFTR protein that regulate expression, maturation, poly‐ubiquitination, and trafficking of CFTR and subsequently, its role in inflammatory responses. More specifically, K14/K68/K1218 residues are important for lysosomal degradation, while the K710/K716/K1080 residues are necessary for stabilization/poly‐ubiquitination of CFTR, and the K1041 residue is critical for overall expression of CFTR. Moreover, the K710R, K716R and K710/716R mutants are inflammatory however, inhibition of lysosomal function dampens inflammatory signaling by these mutations. K14R, K68R, K1041R, K1080R and K1218R mutants are not inflammatory. Inhibition of proteasomal degradation enhances non‐inflammatory effect of K14R and K1080R mutants.ConclusionsPost‐translational modifications in CFTR lysine residues regulate maturation and trafficking of CFTR and the inflammatory responses.Supported by R01 HL 59410; Eudowood Division of Pediatric Respiratory Sciences
The Brain‐specific BNIP‐2 homology protein Caytaxin acts as a scaffold to transport ATP Citrate Lyase on kinesin and induce neurite outgrowth
Authors: Sun Jichao , Low Boon Chuan 
Published: June 2021

Mutations in the gene, ATCAY that encodes the brain‐specific protein Caytaxin, is linked to the development of the human disease, Cayman ataxia. To understand the cellular and molecular mechanisms underlying the neuronal function of Caytaxin, proteomics pull‐down was performed and ATP citrate lyase (ACL) was identified as a novel interacting partner of Caytaxin. In rat pheochromocytoma PC12 cell, Caytaxin and ACL overexpression enhanced neuronal development characterized by long protruding neurites. However, deficiency of either Caytaxin or ACL by genetic knockdown led to marked reduction in neurite outgrowth, despite the presence of the binding partners. Caytaxin requires ACL enzyme activity to promote neurite outgrowth. Further biochemical and live imaging studies revealed that Caytaxin functions as a scaffold protein that links ACL and kinesin light chain motor protein without altering the ACL activity, and that enhances ACL trafficking along the neurites. Our results therefore support the notion that Caytaxin could regulate neuronal development through its binding and trafficking of ACL. Consequently, loss of Caytaxin function could affect ACL signaling pathway, leading to ataxia.This work is supported in part by grants from the Biomedical Research Council of Singapore and Mechanobiology Institute, co‐funded by National Research Foundation and Ministry of Education.