bluuebunny/crossref_metadata_2025_split huggingface dataset:
How arrestin assembles MAP kinases into a signaling complex
Authors: Song Xiufeng , Gurevich Vsevolod V 
Published: June 2021

Arrestins interact with GPCRs and numerous non‐receptor partners. We found that all four members of the arrestin family bind protein kinases JNK3a2 and ASK1, but only arrestin3 facilitates JNK3 phosphorylation. Mutant arrestin which does not bind receptors induced even greater JNK3 activation, demonstrating that this is a function of free arrestin. In contrast to arrestin2, arrestin3 has a native nuclear export signal (NES) and is exclusively cytoplamic. We explored the role of subcellular localization and found that arrestin3 with disabled NES activates JNK3 less efficiently. However, even engineered NES+ arrestin2 does not activate JNK3 as strongly as arrestin3. Truncated JNK3 lacking both N‐ and C‐termini does not bind arrestin3. The deletion of only N‐ or C‐terminus of JNK3 reduced binding to arrestin3 compared to full‐length, suggesting that both termini mediate this interaction. To identify arrestin3 elements involved in JNK3 and ASK1 binding, we used nuclear exclusion assay and coimmunoprecipitation. Separately expressed N‐ and C‐domains of arrestin3 interact with both JNK3 and ASK1, demonstrating that each kinase has two binding sites on arrestin. These data suggest a novel model of arrestin‐dependent scaffolding of MAP kinase cascades: arrestin3 assembles the components with both MAPKKK and MAPK positioned along the long axis of the arrestin molecule.NIH grants GM 77561, EY11500 (VVG)
The Synthesis of Polypeptide Analogues used as Molecular Transporters
Authors: Roper Latoya A. , Olson David , Danowski Barbara , Kehlbeck Joanne D. 
Published: June 2021

Molecular transporters are compounds noted for their unique ability to traverse the cell membrane while transporting cargo into the cell. One such macromolecule studied is the naturally occurring HIV‐1 transactivator of transcription (TAT). A nine amino acid sequence rich in arginine residues is responsible for cellular uptake (R49KKRRQRRR57). According to Rothbard and colleagues, the polarizability and potential for bidentate hydrogen bonding of arginine with cell surface proteoglycans like heparin sulfate allows for effective cell penetration.To test this proposal will we have synthesized polypeptide analogues called polypeptoids. Peptoids are constitutional isomers of peptides and differ in the connectivity of the side chain. While the side chain of the amino acid residue in a peptide is attached to the alpha carbon, the side chain of a peptoid is attached to the nitrogen. Polypeptoids serve as excellent mimics of polypetides and are stable to proteolysis. In particular the synthesis of a novel peptoid compound containing guanine group in the side chain will be described. The side chains of this molecule have increased polarizability and the potential to participate in bidentate hydrogen bonding. The cellular uptake properties of this compound will be compared to that of polypeptoids with arginine‐like side chains in live cell assays.Funding for research provided by Union College Chemistry Department and Union College Future Professors Program.
Role of Human Skin Resident T Cells in Wound Healing
Authors: Toulon Antoine , Jameson Julie , Bhavsar Dhaval , Tenenhaus Mayer , Rudolph Ross , Gueniche Audrey , Breton Lionel , Havran Wendy 
Published: June 2021

Cutaneous wound healing is a complex process requiring constant communication between cells in the form of cytokine release, cell‐to‐cell contacts and cell‐to‐matrix interactions. Chronic wounds are a serious clinical problem affecting numbers of elderly and diabetic patients. Skin γδ T cells have been shown to play unique roles in tissue homeostasis and during tissue repair factors in mice. Since the distribution of epidermal resident T cells (γδ vs αβ) is different in mice and humans, we aimed to characterize the role of epithelial resident T cells in wound repair in humans. We investigated the ability of T cells isolated from normal epidermis, acute wounded skin and chronic wounds to secrete Insulin‐like Growth Factor‐1 (IGF‐1), a growth factor that has been shown to be critical for wound healing. We found that epidermal resident T cells freshly isolated from normal skin produce IGF‐1 upon in vitro stimulation. Interestingly both αβ and γδ T cells were activated in acute wounds and secreted IGF‐ at an early stage of the wound healing process. In contrast, in chronic wounds neither T cell subset produced IGF‐1 or was able to respond to in vitro stimulation, supporting a crucial role for human skin T cells in tissue repair. Further characterization of the functional activity of resident T lymphocytes in acute wounds should allow the development of new therapeutic strategies to treat chronic wounds.Funded by L’Oreal and NIH AI36964
The role of glucose energy metabolism in B lymphocyte survival responses
Authors: Dufort Fay J. , Blair Derek , Bleiman Blair , Gumina Maria R. , Chiles Thomas C. 
Published: June 2021

Naïve B lymphocytes must acquire and metabolize external glucose in order to support the bioenergetics associated with maintaining cell volume, ion gradients, and basal macromolecular synthesis. Interleukin‐4 (IL‐4) is a pro‐survival cytokine that prevents death of naïve B lymphocytes, in part, through the up‐regulation of anti‐apoptotic proteins, such as Bcl‐xL. We show that IL‐4 stimulates glucose utilization in the form of glucose uptake and glycolytic catabolism in ex vivo splenic B cells; this is accompanied by increased cellular ATP. IL‐4‐induced glycolysis occurs normally in B cells deficient in insulin receptor substrate‐2/phosphatidylinositol 3‐kinase signaling. By contrast, Stat6‐deficient B cells exhibit impaired IL‐4‐induced Glut1 expression and glycolysis. In keeping with this, inhibition of B‐cell antigen receptor signaling following co‐cross‐linking of FcgammaRIIb results in decreased glucose uptake and glycolytic metabolism and lower cellular ATP. Moreover, mitochondrial homeostasis is significantly impaired as evidenced by decreased oxygen consumption and FcgammaRIIb‐induced mitochondrial membrane depolarization. These results suggest that glycolysis is necessary to maintain cell survival in either naïve B lymphocytes or in response to negative signaling by the FcgammaRIIb.This work was supported by USPHS grant AI60896 and AI49994.
Listeriolysin O (LLO) as a Vaccine vehicle
Authors: Vivanco Hector , Unanue Emil R. 
Published: June 2021

Specific Aim: In this project we aimed to test antigen presentation to Listeriolysin O (LLO), the major virulence factor of Listeria monocytogenes (LM). The purpose was to examine the immunogenicity of LLO and its possible use as a carrier of antigens for immunization purposes.ResultsPeptides were made to segments of LLO molecule known to induce CD4 responses. C57Bl/6 mice were immunized with the peptides and their CD4 T cells were isolated, made into hybridomas and used to examine the presentation of LLO ex vivo. The CD4 T cell response was extraordinarily potent when LLO was used as an exogenous antigen testing different Antigen presenting cells (APC). The response to LLO was about 10,000 times more efficient than that for peptide. Looking for a safer molecule without the detrimental effects at higher concentrations, we generated a LLO mutant, the LLOW492A, which has a 90% reduced lytic activity but still maintains the binding capacity to the cell membranes. When we tested this LLO mutant ex vivo the magnitude of the response at low doses of antigen was as strong as that for the wild type LLO. We generated a recombinant LLO protein by attaching a CD4 epitope to the NH2 terminus region. We used a known epitope (45–65 epitope) of the hen egg white lysozyme (HEL) molecule. When we tested the response of a known T cell to 45–65 epitope of HEL linked to the amino terminus region of LLO, the response was about 100 fold stronger than to the peptide alone.Conclusions: We have identified an extraordinarily potent LLO‐specific CD4 T cell response when the whole molecule or a mutant derivative is used as an exogenous antigen. Peptides linked to the protein are likewise highly immunogenic. LLO is a potentially powerful vehicle for immunization.
Cholesterol regulates the loading of foreign antigens onto MHC class II in dendritic cells
Authors: Berger Adam Craig , Roche Paul A. 
Published: June 2021

In immature DCs, MHCII resides in cholesterol‐rich intracellular lysosomal compartments. Upon activation, intracellular MHCII binds antigenic peptides and traffics to the plasma membrane. In this study, we have investigated the role of membrane cholesterol in MHCII transport through the use of the cholesterol transport inhibitor U18666A. Pretreatment of immature DCs with U18666A had no effect on the LPS‐induced expression of either the DC maturation marker CD40 or cellular levels of total MHCII and only a slight effect on CD86. However, U18666A treated DCs had a reduced capacity to stimulate naïve antigen‐specific T cells. Similar results were found using DCs from mice lacking NPC1, a protein involved in lysosomal cholesterol egress. Pretreatment of DCs with U18666A resulted in a 4‐fold reduction in the surface and cellular expression of MHCII‐peptide complexes in immature DCs and almost completely inhibited the LPS‐induced appearance of MHCII‐peptide at the plasma membrane of mature DCs. Microscopic analysis of U18666A treated DCs shows that MHCII‐peptide complexes are not found in cholesterol engorged lysosomal antigen processing compartments, suggesting that there are fewer compartments that are competent for antigen loading. We conclude that lysosomal cholesterol regulates MHCII‐peptide loading but not MHCII transport from lysosomes to the plasma membrane in DCs.This work is supported by the NCI.
Maturation and Activation of Antigen Presenting Cells in Young and Aged Mice by a Novel Adjuvant Derived From Human Anaphylatoxin C5a
Authors: Morgan Brandon N , Morgan Edward L , Sanderson Sam , Phillips Joy A , Thoman Marilyn L 
Published: June 2021

Maturation and Activation of Antigen Presenting Cells in Young and Aged Mice by a Novel Adjuvant Derived From Human Anaphylatoxin C5aHuman anaphylatoxin C5a is known to enhance humoral and T cell mediated immunity and inflammation. We have developed an analog peptide based on the C‐terminal region of human C5a [EP67: YSFKDMP(MeL)aR which preferentially enhances immune function. We hypothesize that enhanced immune function occurs via the recruitment and activation of antigen presenting cells. EP67 induces the activation and maturation of splenic adherent and bone marrow derived cells from both aged and young mice in vitro. Supernatants derived from these cultures contained IL‐6, TNF‐α, and INF‐γ, representing a pro‐inflammatory cytokine profile. EP67 also promotes the expansion and maturation of CD11b+, CD11b+ CD11x+, and CD11b+ CD11c+ CD80+ CD86+ cells derived from young and aged bone marrow. These results suggest that these adjuvant peptides promote the activation of antigen presenting cells in the enhancement of humoral and cell‐mediated immune function. This work was supported by Grants AG09948–09, AG031496, and AI065712
A method for enrichment and detection of viable mouse IL‐17‐secreting T cells
Authors: Richter Anne , Foerster Anna , Assenmacher Mario 
Published: June 2021

IL‐17 is discussed to be a key player in the protection against certain infections, but it is also involved in the pathogenesis of autoimmune diseases. We have developed an IL‐17 secretion assay, a tool for the detection and enrichment of viable mouse IL‐17‐producing cells.Mouse T cells were either directly ex vivo activated with PMA/ionomycin or beforehand stimulated and expanded using CD3/CD28 antibodies. Secreted IL‐17 was detected on the surface of IL‐17‐producing cells after labeling first the cells with a bispecific CD45‐IL‐17 antibody affinity matrix. During culturing IL‐17 was secreted and specifically caught by the affinity matrix of IL‐17‐producing cells. Subsequently IL‐17‐secreting cells were fluorescently stained with a second IL‐17 antibody or additionally labeled with magnetic beads. We detected IL‐17 secretion by CD4 T cells, CD8 T cells and γδ T cells. Already after 1–3h of stimulation the frequency of IL‐17‐secreting cells was maximal. Costaining of cells for secreted IL‐17 and intracellular IL‐17 confirms the specificity of our labeling method. After magnetic enrichment we yield populations of IL‐17+ T cells with purities of more than 90–95%.The IL‐17 secretion assay allows the detection and provides highly enriched populations of viable IL‐17‐producing cells to investigate the role of IL‐17 in host defense and disease development.
A small subset of dendritic cells controls the Th2 bias of neonatal immunity through an interplay between IL‐12 cytokine and IL‐13Rα1 expression
Authors: Hoeman Christine Marie , Lee Hyun‐Hee , Hardaway John , Zaghouani Habib 
Published: June 2021

It is known that neonates are susceptible to microbial infection and allergic reactions. This is likely due to the lack of Th1 but excess of Th2 cells. The mechanism underlying this Th1/Th2 unbalance has not been clearly elucidated. When exposed to antigen (Ag) on the day of birth, both Th1 and Th2 cells develop in the primary response but Th1 cells up‐regulate IL‐13Rα1 chain which associates with IL‐4Rα to form an IL‐4Rα/IL‐13Rα1 heteroreceptor. During rechallenge with Ag, IL‐4 from Th2 cells utilizes this heteroreceptor to drive apoptosis of Th1 cells, hence creating a secondary Th2 immunity bias. Herein, it is shown that day 6 after birth represents a point at which the neonate acquires the ability to develop Th1 responses and overcome the Th2 bias. This transition lies on delayed developmental maturation of the CD8α+CD4‐ dendritic cell (DC) subset. Indeed, during the first 6 days of birth the neonates had minimal CD8α+CD4‐ DCs and were unable to produce significant IL‐12 which led to up‐regulation of IL‐13Rα1 expression on primary Th1 cells and formation of IL‐4Rα/IL‐13Rα1 heteroreceptor. By day 6 accumulation of CD8α+CD4‐DCs reaches optimal level leading to significant IL‐12 that opposes IL‐13Rα1 up‐regulation on Th1 cells and minimizes IL‐4Rα/IL‐13Rα1 expression.This research was supported by grants 2RO1AI48541 and R21AI06279 from NIH. CMH is supported by training grant GM08396‐16A1 from NIGMS.
Chemopreventative activity of a cyclooxygenase‐2 inhibitor in FBV transgenic mice with a CMV promoter driven MMTV/neu transgene
Authors: Abe Fuminori , Donkor Moses , Scholar Eric , Dafferner Alicia , Westphal Sherry , Hoke Traci , Talmadge James E. 
Published: June 2021

We have previously examined the effect of transplantable, murine mammary tumor growth on the expansion of myeloid‐derived suppressor cells (MDSCs) and their tumor infiltration, as well as the relationship of MDSC frequency to T cells. We report an extension of these studies using mouse mammary tumor virus (MMTV) promoter driven HER2/Neu proto‐oncogene transgenic, FBV mice (neu transgenic mice). Studies in this model also included an assessment of the tumor chemopreventative activity by the cyclooxygenase‐2 inhibitor, celecoxib (80 mg/kg in drinking water). We report that celecoxib administration delayed tumor induction, increased the incidence of apoptotic tumor cells in the treated mice, and prolonged survival. In addition, immune surrogates in the spleens and tumors were studied by flow cytometry and quantitative reverse transcriptase polymerase chain reaction analysis. These studies revealed that tumor development expanded MDSCs numbers and depressed the T cell frequency in the spleen. Celecoxib administration reduced the MDSC expansion and T cell depression in the spleen and tumors. The tumor associated increase in transcript levels of COX‐2, G‐CSF, GM‐CSF TGF‐β, NOS‐2, ARG‐1 and VEGF‐α were reduced by celecoxib administration. The greatest increase in cytokine and enzyme levels were observed within tumor infiltrating leukocytes relative to spleen cells, suggesting a tumor‐immune cell interaction.Supported in part by funding from NIH grant 5 RO1 AT001739, Eppley Cancer Center and Nebraska DHHS LB506.
Access of Peruvian families to animal source foods when their child is 6–18 months and height for age at 4–5 years old
Authors: Penny Mary Edith , Marin R Margot 
Published: June 2021

Micronutrient deficiencies are a cause of stunting in young children. Animal source foods (ASF) are good sources of bioavailable micronutrients and interventions promote access to ASF. We used data from 1954 families in the YoungLives longitudinal study of Peruvian children to examine the association of family access to ASF when the child was 6–18m (1st round data collection) and height‐for‐age at 4–5 yrs (2nd round). Families reported the value of bought and home grown foods for 32 different food groups including 5 ASF groups for a 2 week period. Results are reported as $US/month. 97.6% of families consumed animal source foods with a median per capita (<5yrs old) value of $6.6/m, range 0–60.7. Univariate analysis explored the association of Z‐score height‐for‐age with maternal education and height, wealth index, district of residence, urban/rural, crowding, number in household, value of food eaten, and birth weight. A generalized linear model including ASF per capita consumption,urban/rural and district explained 28.4% of the variability in height‐for‐age but there was colinearity with maternal education, and a model substituting this for ASF consumption, explained 29% of the variability. The influence of ASF access in early childhood on later height varied by district and could not be differentiated from maternal education level.YoungLives is funded by the UK Department for International development.
Family access to animal source foods:an analysis of the Young Lives project in Peru
Authors: Penny Mary Edith , Marin R Margot , Quispe Rosario E , Alarcon Roxana 
Published: June 2021

Animal source foods (ASF) are important sources of bioavailable micronutrients often lacking in the diets of young children. Young Lives is an international study of children growing up in poverty. The monetary value of 32 food groups bought or home produced and eaten during the preceeding 2 weeks was reported by 2049 families with children 6–18m in 20 districts of Peru. Cost was converted to US$/month. Median family consumption of food was valued at $82.6/m. 77.7% of food was purchased. Home production consisted mainly of grains, tubers, and pulses, all red meat, poultry and fish was purchased. Only 2.4% of households reported no ASF but the median per capita, (children and adults >5yrs) expenditure varied from $0–$60.7/m, median $6.6/m. Per capita, urban families spent median $8.1/m, range $0–$60.7; rural families median $4.2/m, range $0–$45.1. Eggs were the most common reported ASF, consumed by 90.1% of urban (U) and 81% or rural (R) families. For other groups of ASF % consumption was: Red meat U 71.2%, R 55.5% rural; poultry U 83.3%, R 49.9%, fish U 66.5%, R 31.6%, cheese, yogurt, butter urban U 60.4%, R 42.7%. The value of animal source foods eaten was associated with total value of food acquired, educational level of the mother and wealth index. Most families had access to one or other animal source foods but these were often bought in small amounts.YoungLives is funded by the UK Department for International development
The consumption of sugar‐sweetened beverages (SSBs) is associated with the increment of body mass index (BMI), mass fat proportion (MFP) and waist circumference (WC) in Mexican adolescents
Authors: Denova Edgar , Jimenez Alejandra , Huitrón Gerardo , Salmerón Jorge 
Published: June 2021

The objective was to evaluate the relation between SSBs consumption and body mass index (BMI), mass fat proportion (MFP) and waist circumference (WC) in Mexican adolescents.A transversal analysis using data from adolescents from Mexico was performed. Information gathering was carried out from 2004 to 2006 by means of a self‐applicable survey. SSBs consumption was evaluated through a semi‐quantitative survey on food consumption frequency. BMI was calculated as weight in kg/height in mt squared, at the time MFP and WC using standardized techniques. Information on socio‐demographic conditions was obtained. The relation was evaluated by means of models of multiple linear regression and logistical regression.A total of 1055 adolescent from 10 to 19 years of age (14.5±2.5) were evaluated. Those individuals with a consumption of 3 o more portions of SSBs a day increased in 0.8 kg/m2 their BMI (p=0.04), independently from other variables. We also observed that those subjects with consumptions of 3 or more proportions of SSBs have a risk 1.7 times superior to suffer from central obesity in respect to those who consume < 1 SSBs proportion/day (p=0.02).Our data corroborate the hypothesis that the consumption of SSBs is associated with an increment in the risk of overweight and obesity in adolescents, as well as an increment in MFP and WC.Financing by Autonomous University of the State of Mexico.
Selective, Multi‐Target Protein Kinase Inhibitors, Rho iso‐alpha acids (RIAA) and Proanthocyanidins, Combined with Cholesterol Lowering Foods, Reduced Cardiovascular Risk Factors in Humans with Metabolic Syndrome
Authors: Tripp Matthew , Lerman Robert , Lamb Joseph , Minich Deanna , Darland Gary , Konda Veera , Emma Dennis , Carroll Brian , Babish John , Pacioretty Linda , Bland Jeffrey 
Published: June 2021

Background: Metabolic Syndrome (MetS), insulin resistance and hypercholesterolemia are major cardiovascular disease (CVD) risk factors. Epidemiological studies suggest that a phytochemically poor diet may be implicated, however, the effect of specific dietary phytochemicals on MetS remains largely unexplored.Objective: To identify phytochemicals which improve insulin sensitivity using in vitro models, and investigate their effects in animal models of insulin resistance, and on CVD risk factors in humans with MetS.Results: RIAA and proanthocyanidins improved insulin signaling in an in vitro 3T3L1 adipocyte screen. In cell free assays they were found to be selective, multi‐target protein kinase inhibitors (e.g., GSK3α and β; PKCβ2; IKKβ), possibly explaining their mode of action. In combination they reduced insulin and glucose to a similar degree as metformin in obese db/db mice. Thus, a human clinical study was conducted. In this 2‐arm, open‐label, 12 wk study, volunteers with MetS and hypercholesterolemia were randomized to either a phytochemical rich Mediterranean‐style, low glycemic load diet, MED (n=19) or MED plus tailored phytochemicals (PED) (n=25) including soy protein, plant sterols, RIAA, and proanthocyanidins. Phytochemical supplementation (PED) led to a greater decrease in CVD risk compared with diet alone (P<0.001), a result independent of weight loss or caloric restriction.