bluuebunny/crossref_metadata_2025_split huggingface dataset:
Engagement of TLR2 during priming directs enhanced CD4 T cell expansion and protective capacities compared to other TLR agonists
Authors: Chandran Smita Sarat , Teijaro John R , Fenton Matthew , Farber Donna L 
Published: June 2021

Toll‐like receptors (TLR) comprise a family of pattern recognition receptors that recognize pathogen‐associated motifs, and trigger antigen‐presenting cell (APC) activation for priming CD4 T cells. We hypothesized that CD4 T cell responses could be optimized by targeting engagement of specific TLR during priming. We used an in vivo system in which HA‐specific or OVA‐specific CD4 T cells are CFSE labeled, adoptively transferred into BALB/c hosts, and subsequently primed with antigenic peptide in the presence/absence of agonists for TLR2 (Pam3Cys), TLR3 (polyI:C) or TLR4 (lipopolysaccharide). CD4 T cells were harvested 1–2 weeks post‐priming to monitor expansion, functional capacity and protective responses against PR8 Influenza challenge. Transferred CD4 T cells underwent robust antigen‐driven proliferation and expansion when primed with both peptide and TLR agonist compared to peptide alone. Pam3Cys‐mediated priming resulted in significantly expanded populations with high levels of IFN‐γ and IL‐2 production, whereas TLR3 and TLR4 engagement resulted in predominant IFN‐γ production with low IL‐2 production. When challenged with a sub‐lethal dose of Influenza virus, Pam3Cys‐primed CD4 T cells afforded better protection in terms of lung function and viral load compared to TLR3 and TLR4 agonists. Thus targeting TLR2 during early priming leads to enhanced CD4 T cell activation, function and protection.Supported by NIH grants R21AI065950 and RO1AI50632
Crosstalk of the Complement and Coagulation System
Authors: Amara Umme , Albers Sonja , Gebhard Florian , Bruckner Uwe B. , Huber‐Lang Markus 
Published: June 2021

Introduction: The complement and coagulation systems are phylogenetically old and important ingredients of innate immune effectors that sense incoming signals of danger. The possible crosstalk and the precise pathways, by which the activation of coagulation triggers the complement cascade and vise versa, remain unclear. Therefore, in the current study the coagulation factors FXIa, FXa, FVIII, FVII, FVIIa, plasminogen, PC, aPC and thrombin were investigated for their potency to in vitro generate C3a and C5a.Material and Methods: In vitro cleavage experiments with incubation of native C3/C5 ± coagulation factors (FXa, FXIa, FIXa, thrombin) ± various serine protease inhibitors (aprotinin, leupeptine, pefabloc, protease inhibitor cocktail) were performed at 37°C dose‐ and time‐dependently by western blots. The enzymatic activity was assessed by ELISA (C3a Quidel REF A015; C5a, DRG Diagnostic REF EIA 3327).Results: This study demonstrates bi‐directional interaction of both cascades as the coagulation factors Xa, Xia, IXa and thrombin can dose‐ and time‐dependently cleave the key proteases of the complement system, C3 and C5, and generate biologically active C5a and C3a, resp, as detected by WB, ELISA and chemotactic activity assays of either HMC‐1 cells or neutrophils. The clotting factor FXIa, FXa, IXa and thrombin‐induced C3 and C5 cleavage activity could dose‐dependently be blocked by the serine protease inhibitor leupeptine and by the structure‐dependent inhibitor pefabloc. Furthermore, FXa exhibited maximal enzymatic activity of all measured coagulation factors (FXa<FXIa<thrombin<FIXa<ctrl) in accordance to the Michaelis‐Menten equation.Conclution: The data provide first evidence of activation of the complement system by various serine proteases of the coagulation system.
Both the lectin and alternative pathways of complement activation play roles in Cryptococcus gattii infection
Authors: Mershon Kileen Louise , Vasuthasawat Alex , Morrison Sherie L. , Beenhouwer David O. 
Published: June 2021

Complement is a cascade of serum proteins involved in opsonization, membrane lysis, and anaphylaxis. Complement can be activated through three pathways: classical, alternative, and lectin. It has been shown that pathogenic Cryptococcus species activate the alternative pathway of the complement cascade, while the polysaccharide capsule blocks activation of the classical pathway. While human mannose binding lectin (MBL) has been reported to not bind to Cryptococcus, no published studies have characterized murine MBL binding to Cryptococcus. Using flow cytometry, we analyzed C3 deposition on C. gattii in the presence of serum from C57BL/6J (intact complement pathways), factor B‐deficient (lacking alternative pathway), and MBL‐deficient mice (lacking lectin pathway), and found that complement activation occurred via both the alternative and the lectin pathways. When mice were infected intravenously with C. gattii, C3‐ and factor B‐deficient mice died significantly faster than C57BL/6J mice and showed higher fungal burdens in the lungs. Radiolabeled heat‐killed C. gattii injected intravenously cleared rapidly in C57BL/6J mice. However, in factor B‐deficient and C3‐deficient mice, organisms cleared much less rapidly, indicating a role for complement activation in organism clearance.NIH R01 AI51415‐5
The CD4+ T cell‐ and SAP‐dependent IgG anti‐polysaccharide response to intact Streptococcus pneumoniae, in contrast to pneumococcal conjugate vaccine, is ICOS‐independent
Authors: Chen Quanyi , Akiba Hisaya , Paton James C. , Cannons Jennifer , Schwartzberg Pamela L. , Snapper Clifford 
Published: June 2021

The IgG murine anti‐polysaccharide (PS) and anti‐protein responses to intact Streptococcus pneumoniae (Pn) are CD4+ T cell and B7/CD28 dependent. However, in contrast to the IgG anti‐protein response, the primary IgG anti‐PS response peaks more rapidly, fails to generate memory, and requires a shorter period of CD4+ T cell help and B7‐dependenrt costimulation. Although CD28 is constitutively expressed on naïve CD4+ T cells, ICOS is typically induced during the first 48h following immunization. We now show that ICOS−/− mice immunized with intact Pn elicit a normal IgM and IgG anti‐PS response but a completely defective anti‐protein response, relative to WT mice. Neutralizing anti‐ICOS‐ligand mAb injected at the time of primary, but not secondary, Pn immunization was inhibitory for the anti‐protein response. In distinct contrast, ICOS−/− mice injected with a pneumococcal conjugate vaccine, exhibited both a defective IgG anti‐PS, as well as IgG anti‐protein response. Although the adaptor molecule, SAP is thought to play a key role in T cell‐dependent humoral immune responses through induction of ICOS on CD4+ T cells, SAP−/− mice challenged with intact Pn were highly defective in eliciting the ICOS‐independent IgG anti‐PS response. These data are the first to demonstrate a CD4+ T cell and SAP‐dependent humoral immune response that is ICOS‐independent.Supported by NIH grant 2R01 AI49192
Effect of Clarithromycin on lung inflammation and alveolar macrophage function in Klebsiella pneumoniae B5055 induced acute lung infection in BALB/c mice
Authors: Kumar Vijay , HARZAI KUSUM , CHHIBBER SANJAY 
Published: June 2021

Objective To evaluate the anti‐inflammatory effect of the clarithromycin as well as its effect on macrophage function in Klebsiella pneumoniae B5055 induced acute lung infection in BALB/c mice.Materials and Methods: Acute lung infection was induced with Klebsiella pneumoniae B5055 by instilling 104 cfu/ml via intranasal route in mice. Group A; control, Group B; clarithromycin (30mg/kg/day/os) treated. The lungs were harvested and homogenized in 1 ml of normal saline and bacterial count, malondialdehyde (MDA), nitric oxide (NO) and myeloperoxidase (MPO) levels. The histopathological examination of the lungs was also done. The lung alveolar macrophage functions (i.e. macrophage spreading phagocytosis) were also assessed.Results: The clarithromycin treated group showed significant (p<0.05) decrease in the neutrophil influx in to the lungs and other markers of inflammation (MDA, NO and MPO) were also significantly reduced (p<0.05). The clarithromycin treatment also significantly (p<0.05) increased the lung alveolar macrophage spreading and their phagocytic potential.Conclusion: On this basis we conclude that clarithromycin decreases the lung inflammation induced by K. pneumoniae B5055 with increasing the macrophage activity against Klebsiella pneumoniae B5055 in the lung tissue.
Heat shock enhances innate immune response to PG‐PS and M. tuberculosis in C. elegans
Authors: Kannan Krishnaswamy , Wan Xiaoxiao , Kannan Aarthi , Fang Deyu , Ortmann Robert 
Published: June 2021

C. elegans is a soil nematode with a functioning immune system. It shares many similarities with mammals in their immune response to various pathogens and serves as a simple in vivo model to study innate immunity, including responses made via the toll‐like receptor (TLR) pathway. We investigated the expression of nlp‐29, a candidate antimicrobial gene, to study the host immune response to two gram positive bacterial isolates: (1) peptidoglycan‐polysaccharide (PG‐PS, an extract from streptococcal cell wall) and (2) the soluble fraction of pulverized heat‐killed M. tuberculosis strain H37ra (M. tb). Neither antigen is pathogenic to C. elegans and the worms do not exhibit any avoidance behavior.Our results showed a differential response to PG‐PS and M. tb, both rich in TLR2 agonists. As compared to OP50 E. Coli fed control, PG‐PS treated worms evoked a robust response by both DAF‐16 nuclear translocation and nlp‐29 expression. In comparison, M. tb treated worms exhibited less robust DAF‐16 translocation and no detectable nlp‐29 expression. However, mild heat‐shock at 37°C for an hour enhanced nlp‐29 expression significantly in PG‐PS treated worms and reached detectable levels in the M. tb treated group suggesting a differential response. Our results demonstrate the presence of an inducible innate immune response in C. elegans to PG‐PS and M. tb, both gram positive bacterial preparations rich in TLR2 ligands.
An Information‐Motivation‐Behavioral Skills Model for Child Sweetened Beverage Consumption
Authors: Goodell L. Suzanne , Amico K. Rivet , Pierce Michelle B. , Ferris Ann M. 
Published: June 2021

As childhood overweight rates increase, a critical need exists to understand parental choices impacting their child's caloric intake. One important and often overlooked contributor to a child's caloric intake is sweetened beverage consumption (CSBC). This study sought to apply a well validated theory of health behavior, the Information‐Motivation‐Behavioral Skills (IMB) model, to CSBC. The proposed IMB‐CSBC model hypothesizes that CSBC‐related information, motivation, and behavioral skills are key determinants of CSBC. We evaluated this model with 198 parents of preschoolers via a self‐report IMB‐CSBC survey and home beverage inventory. Using Structural Equation Modeling, we developed a model accounting for 14% of the variation in CSBC. According to the model (see Figure 1), parents’ information and behavioral skills about CSBC negatively related to CSBC. Behavioral skills also mediated a relationship between information and CSBC, as well as motivation and CSBC. This study provides support for the core IMB‐CSBC model constructs. Using this model, we plan to develop interventions for parents targeting areas such as accurately identifying sweetened and unsweetened beverages, learning how to set and implement limits for child beverage consumption, and reducing the impact of TV on child beverage preference.Funded by Storrs Agricultural Experiment Station, USDA National Needs Fellowship and UConn FSNE
Caregivers’ attitudes regarding portion size served to Head Start children
Authors: Goodell L. Suzanne , Tsuei Eugenia , Hughes Sheryl O. , Nicklas Theresa A. 
Published: June 2021

The purpose of this study was to identify caregivers’ attitudes regarding amounts and types of foods served to Head Start preschoolers using qualitative methods. Researchers conducted 8 focus groups (4 African American; 4 Hispanic) with 33African American and 29 Hispanic Head Start caregivers. Moderator guide questions were based on the Knowledge, Attitudes, and Behavior Model articulated to portion sizes served to Head Start preschoolers. Analysts coded verbatim transcripts for emergent patterns and discrepant data between ethnicities. Caregivers identified child preference, exposure, and pickiness, child age and size, and hunger and the home environment as key influencers on the amounts and types of foods served to Head Start children. Caregivers believed they strongly impact child food consumption through role modeling and verbal encouragement. Using the results from this study, researchers plan to establish how Head Start caregivers’ attitudes impact portion sizes and how portion size impacts consumption. Practitioners should be aware of caregivers’ attitudes regarding their influence on child food consumption and teach these caregivers appropriate behavior modeling and affirmation techniques.Funded by: #NIH 1 R01 CA107545 and NIH Ruth L. Kirschstein National Research Service Award.
Randomized controlled trial comparing the bioavailability of vitamin D3 from fortified Cheddar cheese, fortified low‐fat cheese, and supplement
Authors: Wagner Dennis , Sidhom Gloria , Whiting Susan J , Rousseau Derick , Vieth Reinhold 
Published: June 2021

Objective: We compared the bioavailability of vitamin D3 from novel fortified hard cheeses and from vitamin D3 supplement.Methods: 60 adults were randomly given weekly servings of fortified Cheddar cheese (34 g), fortified low‐fat cheese (41 g), or vitamin D3 supplement. These contained 28,000 IU vitamin D3, equivalent to 4,000 IU/d. 20 adults were also randomized to placebo. Bioavailability of vitamin D3 was assessed by comparing the serum 25‐hydroxyvitamin D [25(OH)D] response over 8 weeks.Results: In the placebo group, initial 25(OH)D, 55.0 Â± 25.3 nmol/L (mean Â± SD), declined over the 8‐wk winter protocol, to 50.7 Â± 24.2 nmol/L (paired t test, p=0.046). In the vitamin D3‐treated groups, the mean increases in 25(OH)D from baseline to 8 wk were: 65.3 Â± 24.1 nmol/L (Cheddar cheese), 69.4 Â± 21.7 (low‐fat cheese), 59.3 Â± 23.3 (supplement with food), and 59.3 Â± 19.6 (supplement without food); these changes were significantly different from the placebo group (ANOVA, p<0.0001) but not from one another (ANOVA, p=0.62). Compared with baseline, serum parathyroid hormone decreased significantly with both the fortification (paired t test, p=0.003) and the supplementation (paired t test, p=0.012) strategies.Conclusions: Vitamin D3 is equally bioavailable from fortified hard cheeses and from supplement. Therefore, hard cheese is a viable food for vitamin D fortification, and can help to enhance vitamin D status.
Glycosaminoglycans isolated from fish muscle enhance iron uptake by Caco‐2 cells
Authors: Llopis José Moisés Laparra , Tako Elad , Glahn Raymond P , Miller Dennis D 
Published: June 2021

Numerous attempts have been made to identify the factor in muscle tissue that enhances Fe absorption but the nature of the promoter remains controversial. Previous research suggested that sulfated glycosaminoglycans (GAGs) from fish muscle promote Fe uptake in Caco‐2 cells (1). This study continues that research to confirm that GAGs enhance Fe uptake by Caco‐2 cells.Fish haddock filets were cooked and lyophilized. GAGs were isolated and separated into low and high sulfur fractions (2). Enzymatic digestion with chondroitinase ABC (50 mU/mg, 1h, 37ºC) and reverse phase chromatography (RP‐HPLC) were used to characterize the GAGs. Nonheme Fe was added to the isolated fractions and to whole cooked fish and an in vitro digestion/Caco‐2 cell model was used to assess their effect on Fe uptake (1).RP‐HPLC analyses showed that retention times of the main components in the high sulfur fraction were consistent with chondroitin/dermatan sulfate‐related structures. This fraction increased Fe uptake up to 185% relative to controls exposed to nonheme Fe. Treatment of the isolated fraction with chondroitinase eliminated the enhancing effect. Unfractioned fish caused a 40% higher Fe uptake than caused by GAG isolates.These results indicate that specific GAGs contribute to the enhancing effect of fish on Fe uptake by Caco‐2 cells but that other factors are also involved.
Calculating Estimated Average Requirements (EARs) for zinc (Zn) According to Dietary Phytate
Authors: Hambidge K Michael , Miller Leland V , Westcott Jamie E , Krebs Nancy F 
Published: June 2021

Background: Estimation of the DRIs (dietary reference intakes) for Zn by the Institute of Medicine (IOM, 2001) specifically included calculations only for phytate‐free diets.The objective of this investigation was to apply a mathematical model (J Nutr, 2007) of daily total absorption of Zn (TAZ) as a function of daily total dietary intake of Zn (TDZ) and of phytate (TDP) to derive EARs for Zn in adults consuming various quantities of phytate.Methods: The IOM estimated physiologic requirements for Zn for men (3.8 mg Zn/d) and women (3.3 mg Zn/d) were used for this purpose. An updated version of the model was used to calculate the TDZ and TDP values which produce TAZs equal to these physiological requirements.Results: The relationship between TDZ and TDP to yield a constant TAZ is linear and described by the following equation for the case of TAZ = 3.8 mg/d: TDZ = 0.0093 × TDP + 8.5, where TDZ and TDP are in mg/d. For a TDP of 1000 mg/d, the model predicts an EAR that is approximately double, and for a TDP of 2000 mg/d an EAR approximately triple, the ‘no phytate’ EAR of the IOM.Conclusion: As these and greater intakes of phytate are typical in developing countries, these calculations have major implications for many populations, not only for EARs, but also for RDAs and Upper Limits.Supported by IAEA and MRDDR Center Grant P30 HD004024.
Hypoglycemic effect of chlorella in Goto‐Kakizaki and Wistar rats
Authors: Kim You Jin , Jeong Hyejin , Kim Mi Kyung 
Published: June 2021

The aim of this study was to examine the hypoglycemic effect of chlorella in 6 week‐old type 2 diabetic Goto‐Kakizaki (GK, n=30) and 6 week‐old normal Wistar (n=30) rats. Animals were fed different three experimental diets containing 0%, 3% or 5% (w/w) chlorella for 8 wks. Blood HbA1c and fasting plasma glucagon concentrations were measured. And HOMA‐indexes were calculated using fasting blood glucose and plasma insulin concentrations.In GK rats, fasting plasma glucagon levels in 3% and 5% chlorella groups were significantly lower than control group, and levels of blood HbA1C, fasting glucose, plasma insulin and HOMA‐indexes in 3% and 5% chlorella groups were tended to be lower than control group.In Wistar rats, blood HbA1C, plasma insulin levels and HOMA‐indexes of 5% chlorella group were tend to be higher, and fasting plasma glucagon levels of 5% chlorella group were significantly higher, than control and 3% chlorella groups.In conclusion, the feeding of chlorella was beneficial to improve insulin resistance in type 2 diabetic GK rats and to maintain blood glucose level in normal Wistar rats.*This work was supported by the grants from Daesang.Co.Ltd. and the second stage of Brain Korea 21 project in 2007.
Polyglutamine immunoreactivity is present in FTLD‐U and normal controls
Authors: Herndon Emily Susanne , Hladik Christa , Shang Ping , White Charles L. 
Published: June 2021

1C2 is a monoclonal antibody raised against the N‐terminus of the human TATA‐box binding protein that contains a string of 38 glutamines. We have previously demonstrated 1C2 immunoreactivity in the substantia nigra, locus coeruleus, and globus pallidus of 2 FTLD‐Ub cases. To investigate this further, we stained sections of the midbrain, pons, and globus pallidus of 10 FTLD‐Ub cases and 2 normal controls using 1C2. In all cases 1C2 immunoreactivity was present in neuropil and in the cytoplasm of neurons, microglia, or both. 9 out of 10 FTLD‐U cases demonstrated 1C2 immunoreactivity in the substantia nigra, but only one revealed immunopositivity in the locus coeruleus. Chromogen deposition was independent of neuromelanin distribution. Granular 1C2 immunoreactivity was present within the neuropil or cytoplasm of globus pallidus microglia in all cases. This strong, reproducible, but previously undescribed 1C2 immunoreactivity warrants further assessment of its binding specificity. Alternatively, expanded CAG repeats may be present in selected CNS locations in normal individuals and those without known CAG repeat diseases. In either case, caution is indicated in interpreting 1C2 immunostaining results in diagnostic and pathogenetic studies of possible CAG repeat disorder cases.Supported in part by NIH ADC grant AG12300, the Winspear Family Center for Research on the Neuropathology of Alzheimer Disease, and the McCune Foundation.
Evaluation of a Novel Vibration Perception Threshold Instrument
Authors: May Kelsi J. , Latman Neal S. 
Published: June 2021

Monitoring peripheral nerve pathology is critical in order to prevent loss of limb or even loss of life. Semmes‐Weinstein (SW) Monofilaments have been the gold standard in non‐invasive peripheral nerve sensitivity monitoring for over forty years. Vibration perception threshold (VPT) instruments have recently been developed as an alternative method of testing peripheral sensitivity, but their measurements exhibit a low correlation with results obtained using the SW monofilaments. One possible reason for this low correlation may be that the current VPT instruments stimulate different receptors than the SW Monofilaments. The purpose of this study was to determine whether a novel VPT instrument, designed to stimulate the same receptors as the SW monofilaments, provides a higher correlation.The two instruments used in this study were the standard SW monofilaments and the novel VPT instrument. Each subject was tested for peripheral sensitivity on the fingertips of their non‐dominant hand using both instruments. The 98 subjects tested ranged in age from 22 to 88 years, with 20% male and 80% female.When analyzed, the SW measurements and novel VPT measurements showed a correlation coefficient of 0.67, significantly greater than the current VPT instruments’ typical correlation of 0.39. The reliability of the VPT measurements demonstrated low intra‐user variability, with an average mean deviation of only 0.3 mV.Therefore, this study indicated that the novel VPT Instrument provides measurements of peripheral nerve sensitivity that have a stronger correlation with SW monofilaments than currently available VPT instruments.
Analysis of secondary structures of odorant receptors
Authors: Nguy May , Qiu Rongde , Feng Ying‐Hong 
Published: June 2021

The structure‐function relationship of rhodopsin family GPCRs is well studied whereas little is known about the family of G protein‐coupled odorant receptors (ORs). Here the amino acid sequences and the functional domains of 385 human ORs were analyzed using AlignX, transmembrane (TM) helix prediction, and helix wheel projection. Most importantly, the critical residues, motifs and domains of ORs were examined by comparison with the corresponding ones of the rhodopsin family, especially the bioamine subfamily GPCRs that bind ligands with small size. Our result shows that as many as 59 residues in the TM domains and 33 residues in the non‐TM domains are conserved in >90% of ORs, an increase of 36.1% over 36 residues in the TM domains of the rhodopsin family GPCRs. Interestingly, the distributions of the highly conserved residues are largely distinct between the two family GPCRs, especially, in TM2, TM3, TM4, TM6 and TM7. The TM3 of ORs bearing highly conserved DRY motif for activation of G&#945; proteins was found to be one residue longer than that of the rhodopsin family GPCRs. These results suggest that ORs may have unique, but rather conserved global conformation and utilize structural mechanisms distinct from GPCRs of the rhodopsin family for ligand binding and activation.Supported by ASPET SURF award to MN and NIH grant (HL065492) to YHF