bluuebunny/crossref_metadata_2025 huggingface dataset:
Pulmonary T cells regulate fibrosis via integrins αVβ3 and αVβ5
Authors: Luzina Irina G. , Nacu Natalia , Lockatell Virginia , Highsmith Kendrick , Choi Jung , Todd Nevins W. , Atamas Sergei P.
Published: June 2021

T cells are commonly involved in lung fibrosis, but their mechanistic role is unclear. Lung T cells from patients with scleroderma, idiopathic pulmonary fibrosis, and healthy controls were studied. CCL18 gene delivery in vivo was used to induce selective chemoattraction of T cells to otherwise healthy or bleomycin‐injured mouse lungs. The fibrotic potential of T cells was studied in cell culture. Immunohistologically and flowcytometrically, integrins αVβ3 and αVβ5 were expressed on 32±15% of T cells in patients with fibrosis but not in healthy controls; higher integrin expression levels correlated with worse pulmonary function measures. Levels of mRNAs for these integrins were also increased. Infiltration of T cell in the otherwise healthy mouse lungs was accompanied by integrin expression on T cells, elevated active TGF‐β, and T cell‐dependent collagen accumulation. In contrast, CCL18‐driven infiltration of T cells combined with bleomycin injury showed lack of integrin expression on T cells, lower active TGF‐β, and partial inhibition of collagen accumulation. In vitro overexpression of integrin αVβ3 or αVβ5 on Jurkat cells caused an increase in TGF‐β and collagen in co‐cultures with primary lung fibroblasts. In conclusion, T cells may act pro‐ or antifibrotically, by expressing or not expressing integrins αVβ3 and/or αVβ5, and likely activating or not activating TGF‐β. Support: R01HL074067, VA Merit (SPA).
Role of cyclophilins in acute allergic asthma
Authors: Balsley Molly Ann , Morton Russell A , Okwumabua Ifeanyi , Bukrinsky Michael I , Constant Stephanie L
Published: June 2021

Cyclophilins can function extracellularly as chemotactic agents for leukocytes. Elevated levels of extracellular cyclophilins have been observed in many inflammatory diseases. Previous studies in our laboratory suggest that extracellular cyclophilins contribute to asthmatic lung inflammation, although the mechanism of contribution is unknown. The objectives of the current study were to establish: (1) the cell source of extracellular cyclophilins and when they play a role in inflammation, (2) the impact of blocking cyclophilins using NIM811 (Novartis), a non‐immunosuppressive form of cyclosporine A, and (3) the mechanism(s) by which cyclophilins contribute to inflammation. Using a mouse model of acute allergic asthma, elevated levels of cyclophilins were observed throughout the asthmatic response, potentially from infiltrating leukocytes. NIM811 treatment resulted in a 30–50% reduction in numbers of eosinophils and effector CD4+ T cells within lung tissues and airways, suggesting that extracellular cyclophilins function by promoting leukocyte recruitment. Blocking extracellular cyclophilins may provide a novel approach for reducing the pathology associated with allergic asthma. Funded by NIH AI‐059208.
Regulator of G‐protein signaling (RGS10) regulates microglial response to chronic inflammatory stimuli and impacts dopaminergic neuron survival.
Authors: Lee Jae Kyung , McCoy Melissa K , Harms Ashley S , Ruhn Kelly A , Gold Stephen J , Tansey Malu G
Published: June 2021

Epidemiological studies suggest that chronic use of non‐steroidal anti‐inflammatory drugs (NSAIDs) lowers the incidence of Parkinson's disease (PD) in humans and implicate neuroinflammatory processes in the death of dopamine (DA) neurons. Chronic systemic inflammation in mice deficient for Regulator of G protein signaling‐10 (RGS10), a microglia‐enriched GTPase accelerating protein (GAP) for G alpha subunits that is also expressed in DA neurons, induced degeneration of the nigrostriatal pathway, suggesting a protective role for RGS10. Knockdown of RGS10 alone in the BV2 microglia cell line resulted in upregulation of inflammation‐related genes and overproduction of neurotoxic cytokines, including Tumor Necrosis Factor (TNF) and enhanced the neurotoxic activity of BV2 conditioned media on the dopaminergic cell line MN9D. Importantly, removal of RGS10 in MN9Ds further enhanced their sensitivity to microglia‐derived mediators, including TNF. Given the neuroprotective effects of RGS10 on DA neuron survival, RGS10 emerges as a novel drug target for the treatment of PD.ααααααααβα
Adjuvant abolishes immune regulatory effect mediated by type II NK T cells
Authors: Maricic Igor , Halder Ramesh C. , Kumar Vipin
Published: June 2021

Myelin‐derived sulfatide (3′‐sulfagalactosyl ceramide) is a self‐glycolipid ligand recognized by a subset of CD1d‐restricted type II NKT cells. Here we show that treatment of mice with an immunodominant isoform of sulfatide inhibits the chronic and relapsing course of experimental autoimmune encephalomyelitis (EAE) in SJL/J mice. However the regulatory effect is lost upon administration of sulfatide lipid with complete Freund's adjuvant.

Using PLP139–151/As‐tetramer and intracytoplasmic cytokine staining techniques we have examined the frequency of and cytokine secretion by pathogenic T cells in lymph nodes as well as in CNS. Our data indicate an increased number of PLP‐reactive Th1‐like cells and enhanced microglial activation in mice co‐challenged with lipid and adjuvant. These results indicate that activation of type II NK T cells in the presence of strong adjuvant is ineffective in the treatment of T cell‐mediated autoimmune diseases. (Supported by grants from the NIH, MSNRC to VK).
The histone deacetylase inhibitor trichostatin A (TSA) upregulates regulatory T cells and modulates renal disease in NZB/NZW F1 mice
Authors: Thomas Megan Ryan , Reilly Christopher , Mishra Nilamadhab , Radjavi Ali
Published: June 2021

In our present studies we sought to determine if the histone deacetylase inhibitor (HDI), trichostatin A (TSA), would alter systemic lupus erythematosus (SLE) in NZB/W mice by regulating CD4+CD25+ T‐regulatory cells. Fourteen to sixteen‐week‐old female NZB/W mice were injected with TSA (1.0 mg/kg BW) intraperitoneally (i.p.) daily, anti‐CD25 (250 mg/mouse) i.p. weekly, both TSA (1.0 mg/kg BW) and anti‐CD25 (250 mg/mouse) i.p. bi‐weekly, or untreated. The mice were monitored for various disease parameters as they aged. Mice were sacrificed at either 26 or 36 weeks of age, tissues collected and assessed for various parameters of disease. Flow cytometric analysis of splenic tissue showed TSA treated mice exhibited increased FOXP3 expression and increased CD4+CD25+ T‐regulatory cells. At 36 weeks, TSA treated animals had decreased anti‐double stranded DNA autoantibodies and decreased protein excretion. Renal pathology showed a decrease in IgG and C3 deposition in renal tissue and a decrease in pathologic glomerular disease in the TSA treated mice. These data indicate TSA decreases lupus nephritis by upregulating FOXP3 expression and CD4+CD25+T cells in NZB/W mice. Taken together, these data show TSA administration modulates lupus in part by increasing T‐regulatory cell function, and warrants further investigation as to its potential in treating SLE.
Methanol extract of saffron ameliorates experimental autoimmune encephalomyelitis in C57BL/6 mice
Authors: Rostami Parvin , Eslambolchi Shamila , Haghbin Kamahldin , Sanati Mohamad H.
Published: June 2021

Experimental autoimmune encephalomyelitis (EAE) is an inflammatory demyelinating disease of the central nervous system, and is commonly used as an animal model for multiple sclerosis (MS). Inflammation is a key event in both EAE and MS, which leads to demyelination and subsequent disability. It is shown that crocus sativus L. stigma (saffron) extracts have anti‐inflammatory activity in rats. There is modern pharmacological evidence for anti‐tumour, chemopreventive and genoprotective effects as well as radical scavenger properties of saffron extracts or its constituents. We observed that both therapeutic doses of dexamethasone and methanol extracts of saffron stigma at 100mg/kg and higher prevented EAE from progressing significantly in pMOG‐immunized female C57BL/6 mice. In contrast, an aqueous maceration extract of Crocus sativus L. did not prevent EAE progression. These results suggest that saffron may attenuate symptoms of EAE by its anti‐inflammatory activity.
Modulation of the Immune Response is Skin Deep
Authors: Bynoe Margaret S.
Published: June 2021

Multiple sclerosis (MS) is an inflammatory attack on the central nervous system (CNS) that results in extensive demyelination and loss of neuronal function that often leads to death. Experimental autoimmune encephalomyelitis (EAE) is the animal model for MS. We have recently observed that when we epicutaneously immunize (on shaved skin) C57BL/6 mice with the autoantigen myelin oligodendrocyte gylcpprotein (MOG), in a skin patch prior to inducing EAE with MOG that these mice are protected from developing EAE. Most importantly, when we epicutaneously immunize mice with MOG after they develop EAE, mice with mild EAE are protected, but not mice with severe EAE. We observed that CD4 T cells from mice that are protected from EAE produced high levels of IL‐10 and they can transfer protection to naïve recipient mice. We also found that epidermal dendritic cells that captured antigen from the patch can suppress EAE when they are isolated after epicutaneous immunization and transferred into naïve recipient mice. These studies will serve to advance our knowledge of autoimmune disease regulation both at the peripheral level and at the CNS level and could be beneficial to MS and other autoimmune diseases.
A Rapid New Procedure for Basophil Isolation from Human Peripheral Blood
Authors: Fairhurst Maureen , Nauer Melany , Eaves Allen , Thomas Terry
Published: June 2021

Basophil research has been limited by the difficulty in isolating pure basophils in large numbers. Typically, basophils comprise < 1% of peripheral blood leucocytes in non‐allergic, healthy humans. Current protocols for their isolation are time consuming involving multiple steps and often special equipment. Prolonged manipulation of basophils can lead to undesirable activation and spontaneous histamine release.

We describe a rapid (within 1.5 hours) and simple method for the enrichment of basophils from normal blood that does not require Ficoll or lysis steps and yields good purity and recovery. Blood was collected with heparin and red blood cells were removed by HetaSep sedimention. The basophils were then enriched using immunomagnetic, column‐free negative selection (EasySep®). Briefly, unwanted cells were specifically labeled with dextran‐coated magnetic particles using a cocktail of bispecific tetrameric antibody complexes. The labeled cells were then removed using a magnet leaving unlabeled basophils. The entire separation procedure can be automated with a pipetting robot (RoboSep®).

Final basophil purity range as assessed by flow cytometry was 92–99% CD45+IgE+ CD123+. May‐Grunwald stained cytospins showed typical morphology and characteristic blue‐black granules. Expression of cell‐surface CD203c was similar between whole blood and enriched basophils. Recovery averaged 49 ± 17 % (n=7).
Protection of IgE‐mediated allergic sensitization by active immunization with antigenized IgE B‐cell epitope in GFP protein scaffold
Authors: Chen Swey‐Shen , Barankiewicz Teresa J. , Yang Yong‐Min
Published: June 2021

Green fluorescent protein (GFP) exhibits a rigid central β‐barrel, formed by eleven β‐strands with floppy loops spanning between the stands. The spanning loop, the site 6 of GFP, was engineered with RE cloning sites for inserting oligonucleotides corresponding to FcεRI‐binding sequence of human IgE. In a high throughput format, oligonucleotides encoding eight amino acids of the receptor‐binding regions were inserted in an expression cassette was assembled with 5’ in vitro transcription and translation enabling sequences. Antigenized C2‐3 linker (C2‐3L) was shown to react strongly by immunoblotting with polyclonal anti‐IgE under native gel electrophoresis and transfer. Recombinant C2‐3L antigenized GFP, expressed in recombinant vector was purified to homogeneity by metal affinity column, followed by Sephacryl S‐200 high resolution gel filtration. Hyperimmune sera from mice immunized with C2‐3L antigenized GFP exhibit high titers of anti‐IgE reactive with JW8 murine/human chimeric IgE. Further, elevated serum anti‐C2‐3L and affinity pure antibodies effectively inhibits binding of JW8 IgE to recombinant FcεRIα. and desensitizes JW8 to rat RBL‐2H3 transfected with human FcεRIα. This observation raises the distinct possibility that active IgE vaccine may indeed be employed in raising active protective anti‐IgE in allergic patients as an alternative to passive immunization with MAb‐E25 anti‐IgE. Moreover, this antigenized epitope (s) platform may be extended to analyzing specific B‐cell epitopes of other proteins of interest.
Tumor‐educated tolerogenic dendritic cells induce downregulation of CD3ε and apoptosis in T cells via an oxygen‐dependent pathway
Authors: Kuang Dong‐Ming , Wu Changyou , Zheng Limin
Published: June 2021

Defects in the CD3/TCR complex and impairment of T cell function are necessary for evasion of tumor, but the underlying mechanisms are incompletely understood. We found that culture supernatants from several types of solid tumor cells drived human monocytes to become tolerogenic semimature dendritic cells (TDCs). Upon encountering T cells, the TDCs triggered rapid down‐regulation of CD3ε and TCR‐α/β and subsequent apoptosis in autologous T cells. Consistent with these results, accumulation of immunosuppressive DCs coincided with CD3ε down‐regulation and T cell deletion in cancer nests of human tumors. The impaired T cell function was mediated by factor(s) released by live TDCs after direct interaction with lymphocytes. Also, the TDC‐induced effect on T cells was markedly reduced by blocking of NADPH oxidase but not by inhibition of arginase, iNOS, indoleamine 2,3‐dioxygenase (IDO), or IFN‐γ. Moreover, we found that hyaluronan fragments constituted a common factor produced by a variety of human tumors to induce formation of TDCs. These observations indicate that tumor microenvironments, including hyaluronan fragments derived from cancer cells, educate DCs to adopt a semimature phenotype, which in turn aids tumor immune escape by causing defects in the CD3/TCR complex and deletion of T cells.
Influence of virulence attenuation on the efficacy of Listeria monocytogenes as a vaccine vector for stimulating anti‐tumor immunity
Authors: Smithey Megan J , Higgins Darren , Freitag Nancy E , Bouwer H.G. Archie
Published: June 2021

Listeria monocytogenes (Lm) has been extensively studied as a paradigm for stimulating cell mediated immunity. Its’ cytosolic replication niche allows secreted proteins to access the MHC class I pathway, such that Lm infection generates long‐lived memory CD8 T cells. Because of its’ potent adjuvant properties and efficacy at stimulating T‐cell responses, Lm is being studied as a vaccine platform for stimulating anti‐tumor immunity. In this study we compared the therapeutic efficacy of wildtype (WT) and virulence attenuated recombinant Lm strains in stimulating tumor specific CD8 T cells in melanoma‐bearing mice. We evaluated the functional and phenotypic properties of vaccine‐elicited CD8 T cells and assessed the ability of these cells to control the growth of both lung metastases and subcutaneous solid tumors. Although assays of CD8 T‐cell effector function revealed comparable stimulation by wildtype vs. attenuated Lm strains, attenuation decreased the ability of the anti‐tumor CD8 T cells to control the outgrowth of subcutaneous solid tumors, with less impact on the control of lung tumor growth. This may reflect a differential effect on regulatory T cells, as only vaccination with WT Lm decreased the Treg frequency at the tumor site. If attenuated vectors are to be pursued, a better understanding of how attenuation influences vaccine efficacy is necessary. This work was supported by VA Merit Review and NIH funds.
Cross‐Talk Between NK Cells and Adenovirally‐Engineered Dendritic Cell Vaccines
Authors: Vujanovic Lazar , Newton Sarah , Landsberg Jennifer C. , Vujanovic Nikola L. , Butterfield Lisa H.
Published: June 2021

Recombinant adenoviral vectors (AdV) are an effective modality for antigen engineering of dendritic cells (DC) in order to harness DC immunostimulatory ability and activate tumor antigen‐specific T cells. Little is known about the capacity of AdV‐transduced DC (AdV/DC) to activate natural killer (NK) cells, and stimulate their cytotoxic and regulatory abilities. Here, we have investigated the effects of AdV transduction on the ability of DC to activate, and become activated by NK cells. We showed that AdV transduction does not enhance DC sensitivity to Granzyme B‐mediated lysis by NK cells when compared to immature DC (iDC). AdV/DC and matured DC both enhance activation of CD56hiCD16− and CD56dimCD16+ NK cell subsets, based on the expression of Granzyme B, CD25, and CD69 activation markers. We detected this effect in short and long‐term cultures, as well as in clinical trial patient PBMC samples post‐AdV/DC vaccination. Reciprocally, NK cells enhance the state of AdV/DC maturation in short‐term co‐cultures according to surface marker expression and cytokine profile evaluation. Moreover, NK cells pre‐activated with AdV/DC kill K562 tumor cells more efficiently than those from iDC co‐culture by Granzyme B ELISPOT. Together, this data demonstrates that there is a positive cross‐talk between AdV/DC and NK cells, which may be an additional pathway by which this vaccine platform may induce anti‐tumor activity.

This work was supported by the University of Pittsburgh Cancer Institute and the Henry L. Hillman Foundation (to L.H.B.)
Stimulation splicing, IRES translation, and intron‐less gene are novel mechanisms in generation of self antigens
Authors: Yang Xiao‐Feng , Yan Yan , Xiong Zeyu , Wang Hong
Published: June 2021

The question of how non‐mutated self‐tumor antigens (>2,000) and autoantigens (>200) gain immunogenicity remained poorly defined. To address this issue, we first identified the isoform of tumor antigen CML66, CML66L, is the immunodominant isoform, suggesting that alternative splicing is an important mechanism in regulation of immunogenicity of the antigen (Yan Y et al, J. Immunol. 2004). We also found that alternative splicing modulates 100% of 45 autoantigens, in contrast to 42% of the rates of alternative splicing for processing randomly‐selected genes. Therefore, we propose a novel model of stimulation‐responsive splicing for generation of untolerized spliced isoforms of self‐antigens (Yang F et al, Clin. Immunol. 2006), which suggests that in response to various stimuli, alternative splicing provides structural basis for expression of untolerized antigen epitopes. Furthermore, we demonstrated that a novel tumor antigen, MPD6, elicits immune responses against myeloid malignancies. MPD6 is encoded by a secondary open reading frame (ORF) of the gene, and its translation is driven by a new internal ribosome entry site (IRES) (Xiong Z et al, J. Immunol. 2006). Finally, we identified a novel tumor antigen, MPD5, which is encoded by an intron‐less gene located in NEK6 gene locus (Xiong Z et al, Intl. J. Immunopathol. Pharmacol. 2007). Taken together, we have identified three novel mechanisms, alternative splicing, IRES‐driven secondary ORF translation, and intron‐less gene transcription, for generation of immunogenicity of self‐tumor antigens and autoantigens. These findings provide novel insights in development of more effective immunotherapy for tumors and autoimmune diseases.
The role of E2A in B cell chronic lymphocytic leukemia
Authors: Kardava Lela , Foon Kenneth A. , Borghesi Lisa
Published: June 2021

E2A is a major regulator of B cell development and function, but nothing is known about the role of this transcription factor in B cell chronic lymphocytic leukemia (B‐CLL), which is a cancer of mature B cell origin. The aim of the present study was to examine the role of E2A in the pathogenesis of CLL. First we studied the expression of E2A in CLL cells and normal human B subsets by intracellular staining and flow cytometry. We show that E2A is normally expressed at high levels in the precursor stage of human B cell development in the bone marrow and is downregulated in mature B subsets. In contrast, E2A expression in CLL cells is upregulated compared with normal mature B subsets. Further we determined whether small interfering RNA (siRNA) mediated inhibition of E2A protein levels restores normal cell cycling or induces apoptosis of CLL cells. We transfected CLL cells with E2A specific siRNA using Nucleofection. We were able successfully perform this approach on primary CLL cells. Then we examined cell cycle status by staining with DyeCycle Violet reagent and the level of apoptosis using Annexin/DAPI staining. Our results showed that E2A downregulation in CLL cells was sufficient to promote cell cycle progression in 50% of cases and induced apoptosis in all cases studied. We conclude that overexpression of E2A is likely to play a pathogenetic role in B‐CLL and may be a potential target in the treatment of this disease. NIH/NIAMS, USIDNET
Homocysteine levels and 10‐year coronary heart disease risk among Cuban‐Americans with and without type 2 diabetes
Authors: Huffman Fatma G. , Zarini Gustavo , Gomez Gianna Perez , Nath Subrata D.
Published: June 2021

We examined the association between homocysteine (HCY) levels with 10‐year coronary heart disease (CHD) risk in 320 Cuban‐Americans with and without type 2 diabetes mellitus (T2DM). Subjects were randomly recruited from Miami‐Dade and Broward Counties, Florida. HCY levels were analyzed from fasting blood samples and CHD risk was determined using ATPIII's 10‐year Risk Calculator (online version). Statistical analyses used SPSS and included descriptive statistics, ANCOVA, correlation, and multiple linear regression. We controlled for age, gender, T2DM status, years with DM, DM medication(s), and BMI. Diabetic subjects had higher CHD risk (10.8%) than non‐diabetics (8.2%), p=.007. Males had higher CHD risk than females, for the entire group and within diabetic and non‐diabetic groups, ps<.001. Subjects with higher HCY levels had higher CHD risk, r=.21, p<.001. However, after we controlled for the variables, this association was no longer significant and age, gender, and T2DM were the strongest predictors of CHD risk. We conclude that Cuban‐Americans with T2DM are at higher CHD risk than non‐diabetics. Although high HCY levels were significantly associated with high CHD risk, once we controlled for other significant factors, HCY levels were no longer able to predict CHD risk.
Effects of routine oral iron supplementation with or without folic acid during pregnancy: an updated systematic review
Authors: Pena‐Rosas Juan Pablo , Viteri Fernando E.
Published: June 2021

Routine antenatal iron (Fe) or iron and folic acid (FeFol) supplementation may improve maternal health and pregnancy outcomes. This review assesses the efficacy, effectiveness and safety of routine daily (D) or intermittent (I) supplementation. The Cochrane Pregnancy and Childbirth Group's Trials Register (October 2007) was searched and relevant organizations were contacted for identification of additional trials. All randomized trials evaluating the effect of routine antenatal D or I Fe or FeFol were selected considering their methodological quality. Primary outcomes included maternal and infant clinical and laboratory outcomes.

Forty‐nine trials, involving 19147 women, were included. The data suggest improved Hb and Fe nutrition compared to non supplemented women. There are no significant differences between D and I in any specific variables except for higher rate of hemoconcentration during 2–3 trimesters in D than in I, and also higher side‐effects in D, except for vomiting that is more common in I. D Fe and FeFol results in slightly higher maternal Hb at term. Iron deficiency and its anemia at term were slightly less likely to occur in women on D Fe than in non supplemented women. Further studies are needed to assess the effects of pre‐conceptional iron and folate status and antenatal Fe or FeFol doses administered either D or I with emphasis on functional and perinatal outcomes.
Fat oxidation tracks with fatty acid availability at low but not high plasma fatty acid concentrations
Authors: Newsom Sean Alec , Schenk Simon , Horowitz Jeffrey F
Published: June 2021

The purpose of this study was to examine the effect of changes in fatty acid (FA) availability on resting FA oxidation and reesterification. We measured whole‐body fat oxidation (indirect calorimetry) and plasma concentrations of FA and glycerol in seven healthy, non‐obese women (BMI: 22.9±1.0 kg/m2) during and after an overnight (16h) lipid and heparin infusion (LIPID; 0.11 g fat/kg/h) compared with an overnight infusion of saline (SALINE). Although LIPID increased plasma FA concentration more than 2‐fold compared with SALINE (1.10±0.16 vs. 0.41±0.09 mM at 0700h the next morning, P<0.05), this augmented FA availability did not increase fat oxidation. The infusions were stopped at 0700h, and by 0900h plasma FA concentration during LIPID dropped to half that found during SALINE (0.20±0.06 vs. 0.43±0.07 mM, P<0.05). Importantly, this fall in plasma FA concentration was paralleled by a lower fat oxidation in LIPID vs. SALINE (2.0±0.6 vs. 4.0±0.5 umol/kg/min, P<0.05). Up to 3h after the infusions the ratio of plasma FA‐to‐glycerol concentration was lower during LIPID vs SALINE (3.76±0.49 vs. 8.02±2.71; P<0.05), suggesting LIPID had a persistent effect on endogenous fat metabolism by increasing FA reesterification. These data indicate that: 1) plasma FA availability influences lipid oxidation at relatively low but not high plasma FA concentrations, and, 2) elevated FA availability may augment intracellular FA reesterification, thereby limiting endogenous release of FA for oxidation and other metabolic fates.
Dietary glycemic index does not influence prooxidant and antioxidant gene expression in adipose tissue of C57BL/6 mice
Authors: Huggins Kevin W. , Colbert Katie E.
Published: June 2021

The objective of this study was to determine whether a high glycemic index (H‐GI) diet compared to a low glycemic index (L‐GI) diet alters prooxidant and antioxidant gene expression in adipose tissue of mice. C57BL/6 mice (n=8/grp) were fed a diet containing either H‐GI starch (100% amylopectin) or L‐GI starch (60% amylose/40% amylopectin) under low fat (LF) or high fat (HF) conditions for 15 weeks. Meal tolerance tests (MTT) were performed to determine blood glucose response to the experimental diets. The expression of the prooxidant enzyme NADPH oxidase (NOX) and antioxidant enzymes glutathione peroxidase (GPx), catalase (CAT), superoxide dismutase (SOD) was determined by real‐time PCR. In both the LF and HF groups, the area under the blood glucose curve for the MTT was significantly greater (P<0.03) in mice fed H‐GI compared to L‐GI. The HF fed mice had a significant increase (P=0.0005) in NOX expression compared to the LF fed mice. There was no significant difference in NOX expression between H‐GI and L‐GI diet groups. HF diet feeding significantly decreased (P=0.004) the expression of GPx compared to LF feeding. There was no significant difference in GPx expression between H‐GI and L‐GI diet groups. The expression of CAT and SOD was not different among any of the diet groups. Diets differing in GI do not influence prooxidant and antioxidant enzyme gene expression in adipose tissue of mice. (Funded by AAES 080‐025)
Effect of Coffee/Tea on Mean Values and Variability of The Glycemic Index of Foods
Authors: Aldughpassi Ahmed , Wolever, DM, PhD Thomas MS
Published: June 2021

There is accumulating evidence that a low glycemic index (GI) diet may reduce the risk of a number of chronic diseases, thus, there is increasing interest in GI in nutrition research. Since numerous methodological factors influence GI determination, valid use of the concept requires accurate and precise methodology. One factor that has been assumed to be important is the type of drink served with test foods. Thus, to see if allowing subjects to drink coffee/tea affected the mean and variation of GI, the GI values of Fruit Leather (FL), and Cheese Puffs (CP) were determined twice in 10 subjects using the FAO/WHO protocol with white bread (WB) as the reference food. In one series subjects could choose either 250 ml coffee or tea with the test foods, while in the other they had 250 ml water as the drink. Coffee/tea increased blood glucose (BG) 30 min after the WB and CP and reduced BG at 120 min compared to water (p<0.05). Similar trends were seen for FL (p>0.05). There were no significant differences between drinks on mean iAUC and mean GI values for all foods (p>0.05). The within‐subject coefficient of variation of iAUC (CV=100 × SD/mean) for the repeated WB tests with coffee/tea, 21±3.0 %, was less than with water, 30±5.3 %, although the difference was not significant. The GI SEM for FL and CP with coffee/tea 5.6 and 8.3, were less than those with water 49.3 and 11.6. These results suggest that coffee/tea do not affect the mean but may improve the precision of GI values.
Hydroxypropyl methylcellulose, a viscous indigestible polysaccharide, reduces adiposity and lowers plasma leptin and resistin concentrations in rats
Authors: Islam Ajmila , Hesslink Robert L. , Gallaher Daniel D.
Published: June 2021

Previous studies in this laboratory have suggested that viscous indigestible polysaccharides reduce adiposity with little or no effect on body weight. To further understand this effect, we have examined the effect of a viscous indigestible polysaccharide, hydroxypropyl methylcellulose (HPMC) on body composition and plasma adipokines in rats. Male Wistar rats were fed either a cellulose based control diet (no viscosity) (n = 12) or HPMC‐containing diet (high viscosity) (n = 24) for 6 weeks. Body composition was measured by dual X‐ray absorptiometry (DEXA) before and after 6 weeks of feeding. Rats fed the HPMC diet had a high intestinal contents viscosity whereas rat fed cellulose diet had no intestinal contents viscosity. After statistically adjusting for slight differences in final body weight, rats fed HPMC had significantly reduced body fat mass and greater lean body mass than the cellulose group, based on DEXA scanning and lower weights of epididymal and perirenal fat pads. Rats fed HPMC had significantly lower fasting plasma leptin and resistin concentrations. However, plasma adiponectin levels did not differ between the groups. There were no significant differences between groups in fasting plasma glucose or insulin levels. These results indicate that viscous indigestible polysaccharides reduce adiposity independent of an effect on body weight. (Supported by Imagenetix Inc.)
Effect of omega‐3 fatty acid suplementation on Omega‐3 Index and red blood cell (RBC) membrane fatty acid composition
Authors: Carughi Arianna , Perelman Dalia
Published: June 2021

Omega‐3 fatty acids (n‐3 FA) have been shown from epidemiological studies and clinical trials to lower incidence of cardiovascular disease (CVD) in patients with pre‐existing CVD as well as in healthy individuals. In randomized secondary prevention trials fish or fish oil have been shown to lower total and coronary heart disease (CHD) mortality at intakes of about 1 g/day. The Omega‐3 Index (EPA + DHA expressed as % of total FA) has been proposed as a physiologically relevant, modifiable, independent and graded risk factor for death from CHD. RBC membrane FA composition correlates well with biomarkers of n‐3 FA including serum EPA and DHA, whole blood EPA, DPA and DHA and FA composition of cardiac tissue. The present dietary intervention study investigates the effect of a fish‐oil based, n‐3 FA supplement on Omega‐3 Index and RBC membrane FA composition. Thirty healthy men and women consumed a supplement providing 1070 mg total n‐3 FA (460 mg DHA, 480 mg EPA, and 80 mg other n‐3 FA) daily for 4 wks. At the end of this period there was a significant 24% increase (p<0.01) in the Omega‐3 Index (Fig.1). While there were no significant changes in % linoleic, gamma linoleic, and alpha linolenic acid; EPA, DPA and DHA significantly (p<0.01) increased compared to baseline values (0.76% versus 1.4%, 2.30% versus 2.58% and 5.49% versus 6.60% respectively), and % arachidonic acid significantly decreased (19.58% versus 18.83%, p<0.05). This study shows that RBC membrane FA composition and so Omega‐3 Index can change in a short time using a fish‐oil based supplement.
Improved bioavailability of calcium in genetically‐modified carrots
Authors: Hawthorne Keli M. , Morris Jay , Hotze Tim , Abrams Steven A. , Hirschi Kendal D.
Published: June 2021

Osteoporosis is one of the world's most prevalent nutritional disorders and inadequate absorbed calcium is a known contributor to the pathophysiology of this condition. In a cross‐over study of 15 male and 15 female young adults, we used a dual stable isotope method with 42Ca‐labeled genetically‐modified (sCAX1) and control carrots and intravenous 46Ca to determine the absorption of calcium from these carrots in humans. Subjects also consumed 48Ca‐labeled milk for comparison. Calcium absorption efficiency was 42.6 +/− 2.8% and 52.1 +/− 3.2% (p<0.001) for the sCAX1 carrots and control carrots, respectively (all data are Mean +/− SEM); however, total calcium absorption per 100 g of carrots was 41 +/− 2% higher in sCAX1 carrots compared to control carrots (26.50 vs. 15.34 mg Ca/100g, p<0.001). Calcium absorption efficiency from the milk was similar to that from the control carrots (50.1 +/− 3.0% vs. 52.1 +/− 3.2%, p=0.7) but was slightly higher than that from the sCAX1 carrots (50.1 +/− 3.0% vs. 42.6 +/− 2.8%, p<0.05). These results demonstrate the first human study to establish the nutritional consequences of a genetically‐modified calcium increase in food and support the use of novel approaches to enhance the micronutrient content of vegetables. Funding: NIH Grants IR01 DK 062366 and USDA CSRESS#2005‐34402‐16401.
Control of Obesity and the Metabolic Syndrome Risk Factors by a Low Calorie‐Moderate Carbohydrates Diet in Mexicans
Authors: de Jesús Vergara‐Jimenez Marcela , Castro‐Acosta Monica Lizzette , Arias‐Gastelum Mayra , Valdez‐Zazueta Guadalupe
Published: June 2021

Obesity (O) is a multifactorial disorder and is associates with the development of several chronic diseases. The metabolic syndrome (MS) is diagnosed when three or more than any factors like dislipidemia, hyperglicemia, hypertension or central obesity are presented. Some studies associated the carbohydrate intake with the development of obesity and the MS has been reported. The objective of this study was to evaluate the effect of a low calorie‐ moderate carbohydrate diet (40% energy) on obesity and the MS in people from Mexico for 8 weeks. 156 subjects participated in the study, however only 29 finished it. Every week, the participants came for the recommended diet used within the intervention and the anthropometric and clinical parameters were measured. Blood sample were drowning at points 0, 4 and 8 weeks of the study which were used to analyze the biochemical parameters. Anthropometric, clinical and biochemical parameters were measured by using standardized methods. Participants lost 14.4 pounds average during the whole intervention, 8.4 cm of waist circumference and 2.5 of BMI. 21.4 and 17.8% reduced hypertension and glucose respectively. 32.1% reduced total cholesterol, 32.1% reduced LDL‐C concentrations and triacylglycerides and 28.6% increased their HDL‐C. Low calorie‐moderate carbohydrate diet (40% energy) is very useful in the control of obesity and the factors related to the MS, and also it resulted very convenient for Mexican since, carbohydrates are an important part of their cultural dietary habits.

FOUNDED BY PROFAPI‐UAS 2006
The addition of docosahexaenoic and arachidonic acid to the diet of artificially reared pups improves the response of splenocytes to lipopolysaccharide
Authors: Goruk Susan , Hoedl Abha , Lau Jenny , Bell Rhonda C , Field Catherine J
Published: June 2021

Immune cells from infants are reported to have a lower cytokine (IFNγ) response to bacterial antigens such as lipopolysaccharide (LPS), which may contribute to their higher susceptibility to infections. Recently, it was reported that the addition of the long chain polyunsaturated fats (PUFA), docosahexaenoic (DHA) and arachidonic (ARA) acid to infant formula reduced the risk of infections. Using the artificially reared rodent model, we studied the effect of feeding isocaloric nutritionally adequate rat milk substitute with or without long chain PUFA (0.24% DHA + 0.36% AA) for one week (12–21 d of age). Both groups of rats grew similar to suckled pups and there was no difference in body or spleen weight. Except for a higher (P<0.05) proportion of dendritic cells (OX62+) in PUFA‐fed rats, supplementation with PUFA had minimal effects on the major lymphocyte phenotypes in spleen. After ex vivo LPS stimulation (48h), isolated splenocytes from the PUFA‐fed rats produced more IL‐1β (1.4X) and IFNγ (1.9X) than those from the control group (P<0.05). These results suggest that adding DHA and AA to the diet of formula‐fed rats significantly improved the immune response to LPS. Funded by NSERC.
Relationship between Glycemic Control and C‐Reactive Protein in Cuban‐Americans with Type 2 Diabetes Mellitus
Authors: Huffman Fatma G , Zarini Gustavo , Swink Michele , Gomez Gianna Perez
Published: June 2021

We investigated the relationship between HbA1c and C reactive protein (CRP) among 182 Cuban‐Americans (Females=110, Males=72) with type 2 diabetes mellitus (T2DM). Subjects were randomly recruited from Miami‐Dade and Broward Counties, Florida. Body mass index (BMI) was calculated from weight and height, and HbA1c and CRP were determined in fasting blood samples. Subjects were stratified into three groups according to HbA1c levels (<7.0%, 7.0–8.9%, and >9.0%). High CRP levels were defined as >3.0 mg/L. Statistical analyses used SPSS and included descriptive statistics, Spearman's rho correlation, and linear and logistic regressions controlling for age, gender, BMI, smoking, years with diabetes, diabetic medications, and NSAID drugs. Subjects with HbA1c >9.0% were significantly younger than those with HbA1c <7.0%, and HbA1c 7–8.9%, p<.05. Subjects with HbA1c <7.0% had significantly less years with T2DM than those with HbA1c 7–8.9%, and >9.0%, p<.05. HbA1c was significantly correlated with CRP, rs=.23, p<.001. For every one unit (%) that HbA1c increased, CRP was predicted to increase by 3.9 mg/L, p<.001. Subjects with HbA1c >9.0% were 3 times more likely to have high CRP levels than those with HbA1c <7.0%, p=.028. Higher HbA1c levels were significantly and positively associated with high CRP levels in Cuban‐Americans with T2DM.
Transposon‐mediated stable integration of genes into migrating cells during blood vessel patterning
Authors: Takahashi Yoshiko
Published: June 2021

A number of cells appear to migrate over long distances in the body during organogenesis. However, the molecular mechanisms by which such migratory behaviors are regulated remain largely unknown. It has been challenging to genetically manipulate and trace migratory cells over a long period of time. Recently, we have developed a Tol2‐transposon‐mediated gene transfer in chickens, a method that allows for stable genomic integration of electroporated genes. Co‐electroporation of a EGFP‐expressing cassette cloned into the Tol2 construct along with a transposase‐encoding plasmid results in persistent expression of EGFP until at least embryonic day 8 (E8), thus surmounting the shortcoming of conventional electroporation techniques that limit the duration of activity of exogenous genes. Using these methods, we have recently found that a subpopulation of somitic cells electroporated with Tol2‐CXCR4 is attracted to forming intercostal vessels, the thin blood vessels protruding from the dorsal aorta, and ultimately differentiates into smooth muscle cells (SMC) around these vessels at E7. The attraction of SMCs is mediated by the chemokine SDF1, which is produced at the tip of the growing vessels. Such selective migration of SMCs appears to determine the patterning of intercostal vessels since these vessels are specifically lacking in CXCR4 KO mice.
ADENOSINE DEAMINASE: AS INDICATOR OF NUTRITIONAL STATUS.
Authors: Feliu Maria Susana , Perris Paula , Slobodianik Nora
Published: June 2021

Previous papers in thymocytes' suspensions of rats, point out to the activity of Adenosine Deaminase(ADA) as potential marker of nutritional status. Moreover, some authors have verified its increase in serum of patients with pathologies related with lymphocytes T. Therefore, subjects with nutritional disorders have shown depression of the immune system, evaluated through immunological markers. So, we analyze if serum ADA activity on populations in nutritional risk, could be considered functional biochemical parameter.

The study included 26 women with anorexia nervosa (AN, 14–32 years, , Body Mass Index (BMI)Kg/m2: 17.9±1.8); 33 obese children of both sexes (O, 5–13 years, , (BMI:Kg/m2: 27.4±4.3) and 16 children with cystic fibrosis (CF, 5 months‐9 years).; the activity of ADA (U/L) was determined by the method of Giusti and Galanti. Groups were compared with the respective age‐match well‐nourished control(C), applying the test of Student. The results expressed as X± SD (UI/L) were: AN: 25.7±8.2*vs. 21.0±4.6(n=22); O: 27.1±9.1*vs. 23.0±5.6(n=48); CF: 36.3±15.8*vs. 23.0±5.6 (n=48)

It was observed an increase in the activity of ADA at a level of p<0.02 in all groups. These findings reinforce the hypothesis to propose the determination of serum ADA activity in the nutrition studies, as a functional indicator related to the mechanisms of defense.

Supported by University of Buenos Aires (B060)
HPLC Methods for Fucoxanthin Analysis in Dietary Supplements
Authors: Craft Neal E.
Published: June 2021

Fucoxanthin (FX) is the second most prominent carotenoid in the earth's biomass. It is the primary carotenoid in some forms of brown algae. FX has been shown to reduce visceral adipose in mice and humans and is being incorporated into dietary supplements for weight loss. To examine the content of commercial FX products, we developed and validated Normal‐Phase and Reversed‐Phase HPLC methods to measure FX. Dietary supplements were purchased from local suppliers and over the internet. Samples were extracted into methanol prior to HPLC analysis. The NPLC was performed on a Diol column using a mobile phase of 6%IPA in hexane while monitoring 450nm. The RPLC was performed on a C30 column using a gradient of methanol, water, and ethyl acetate while monitoring 450nm. Samples tested contained various forms of seaweed or seaweed extracts. Products with stated FX content ranged from 250mcg to 5mg/unit. The products exhibit quite different carotenoid profiles and some do not meet label‐claims for FX. Due to current levels of obesity and increased use of thermogenic supplements, these methods will be useful to monitor the FX in current and future supplements.
Associations between vitamin K and circulating cytokines in older men and women
Authors: Shea M. Kyla , Dallal Gerard E. , Dawson‐Hughes Bess , Ordovas José M. , O’Donnell Christopher J. , Gundberg Caren M. , Peterson James W. , Booth Sarah L.
Published: June 2021

Vitamin K modulates cytokines involved in bone turnover in vitro, suggesting an alternative mechanism by which vitamin K may affect skeletal health. In 379 men and women (60–80y, 59% F) participating in a double‐blind randomized controlled trial (RCT) to assess the effect of phylloquinone supplementation on bone mineral density (BMD), we examined cross‐sectional associations between measures of vitamin K status[plasma phylloquinone and serum % undercarboxylated osteocalcin (%ucOC)] and interleukin‐6 (IL6), osteoprotegerin (OPG), and C‐reactive protein (CRP) concentrations at baseline, and the effect of 3‐years of daily 500mcg phylloquinone supplementation on circulating cytokines. Cross‐sectionally, plasma phylloquinone was inversely associated with IL6 and CRP (p<0.05); %ucOC was positively associated with IL6 (p<0.05). OPG was associated positively with plasma phylloquinone and inversely with %ucOC (p<0.05). IL6, OPG, and CRP concentrations did not differ after 3 years between individuals who received phylloquinone supplementation and those who did not. There was no association between the 3‐year changes in circulating cytokines and BMD. That the baseline cross‐sectional associations between vitamin K status and circulating cytokines were not supported in a 3 year RCT may suggest that vitamin K status is reflective of healthy lifestyle patterns that influence inflammation.
Validation of a semiquantitative food‐frequency questionnaire measuring dietary vitamin K intake in elderly
Authors: Presse Nancy , Shatenstein Bryna , Kergoat Marie‐Jeanne , Ferland Guylaine
Published: June 2021

The study objective was to validate a semiquantitative food frequency questionnaire (FFQ) designed to measure dietary vitamin K (VK) intake specifically. A 50‐item FFQ was interviewer‐administered and compared with data previously obtained from 5‐d food records (FR). Thirty‐nine community‐dwelling healthy men and women aged 65 to 85 years were recruited from the Montréal metropolitan area. Absolute and relative agreements between methods were assessed. Vitamin K intakes measured by the VK‐FFQ (mean ± SD; 222 ± 186 μg/d) were 2.26 times (95%CI = 1.90, 2.67) higher than those obtained by FRs (135 ± 153 μg/d; P< 0.001). However, cross‐classification showed that 72% of participants were correctly classified into thirds and only 8% were grossly classified. Kappa value (κ=0.60) and correlation coefficient (r=0.83) also indicated a good relative agreement. In light of these results, the VK‐FFQ developed is a valid tool for ranking individuals according to their VK intakes. The poor absolute agreement likely results from the inability for FRs to adequately measure the usual intake of episodically consumed foods. The VK‐FFQ will be useful in large‐scale, population‐based research on VK and diseases as well as in clinical practice, especially that focusing on anticoagulant therapy. This research was supported by the Canadian Institutes of Health Research.
Phylloquinone intake is associated with glucose metabolism in middle‐ and older‐aged men and women
Authors: Yoshida Makiko , Booth Sarah L , Meigs James B , Saltzman Edward , Jacques Paul F
Published: June 2021

Animal and metabolic studies suggest that vitamin K may have a beneficial role in glucose homeostasis. The aim of this study was to examine the association between vitamin K intake and measures of glucose metabolism in a community‐based sample of healthy adults. We assessed the cross‐sectional associations between phylloquinone (vitamin K1) intake and glucose metabolism measures in the 2778 participants (1277 men and 1501 women; mean age, 54 y) of the Framingham Offspring Cohort Study. Phylloquinone intakes were assessed by food frequency questionnaire. Surrogate measures of glucose metabolism included fasting and 2‐h post oral glucose tolerance test (OGTT) glucose and insulin, and HbA1c. Associations of glucose metabolism measures with phylloquinone intakes were assessed using the analysis of covariance with Bonferroni adjustment for the multiple outcomes. Lower phylloquinone intake was associated with higher 2‐h glucose and insulin concentrations after adjustment for age, sex, waist circumference, lifestyle characteristics and diet quality (2‐h glucose: 106.4 mg/dL vs. 102.1 mg/dL in the lowest compared to the highest quintile, P‐trend = 0.05; 2‐h insulin: 80.2 μU/mL vs. 72.0 μU/mL, P‐trend = 0.02), but was not associated with fasting glucose and insulin concentrations and HbA1c. Our findings suggest a potential beneficial association of phylloquinone intake with glucose metabolism in the fed state.