bluuebunny/crossref_metadata_2025 huggingface dataset:
Silencing of mitochondrial NADP+‐dependent isocitrate dehydrogenase gene enhances selenite‐induced apoptosis
Authors: Park Jeen‐Woo , Kil In Sup , Chung Kyu Ho
Published: June 2021

Selenium has been shown to play a chemopreventive role in human cancer, presumably by inducing tumor cell apoptosis. Selenite is thought to induce oxidative stress by the generation of the superoxide anion and catalyzing the oxidation of thiol groups. We previously reported that control of the mitochondrial redox balance is a primary function of mitochondrial NADP+‐dependent isocitrate dehydrogenase (IDPm) by supplying NADPH for antioxidant systems. When we investigated whether IDPm would be a vulnerable target of selenite, the loss of enzyme activity was observed. Transfection of HeLa cells with an IDPm small interfering RNA (siRNA) markedly decreased activity of IDPm and enhanced cells?susceptibility of selenite‐induced apoptosis as indicated by morphological evidence of apoptosis, DNA fragmentation, and the modulation of mitochondrial function and apoptotic marker proteins. These results suggest that IDPm siRNA sensitizes HeLa cells to selenite‐induced apoptotic cell death presumably through the perturbation of the cellular redox status.
Expression Analysis of LEDGF/p75 Protein in Major Human Cancers using Immunohistochemistry
Authors: Perez Kayla Y. , Basu Anamika , Casiano Carlos
Published: June 2021

Multiple factors have been associated with increased risk of various cancers including age, diet, race, and inflammatory diseases. Emerging evidence suggests that an augmented state of cellular oxidative stress contributes to progression of various cancers. Our group reported previously that LEDGF/p75, a transcription coactivator, is an autoantigen in prostate, with an elevated expression in advanced stage prostatic adenocarcinomas. Our hypothesis is that LEDGF/p75 might be highly expressed in tissues having high oxidative stress such as tumors, because of its ability to promote survival against cell death induced by oxidative stress, via transcriptional activation of stress and antioxidant genes. Our preliminary screening of LEDGF/p75 in a panel of human cancers has revealed high expression of this protein in ten different tumors. To further validate our findings, we conducted an expression analysis of LEDGF/p75 protein in major human cancers using immunohistochemistry on commercially available human tissue microarrays slides (US BIOMAX), each containing multiple tissues specimens of a specific major tumor type, as well as some non‐cancerous inflammatory disease and normal control tissues. The tumor specimens included seven types of tumors. Paraffin embedded specimens were deparaffinized, rehydrated and stained with either a rabbit anti‐LEDGF/p75 serum using a Biogenic i6000 auto stainer (Biogenex Corp.). Images were acquired using an Olympus BX50 microscope with a Spot RT3TM camera. Normal tissues showed little or no staining. Our analysis revealed high expression of LEDGF/p75 in most of the tumor sections as compared to normal control tissues. A 4‐tier grading system was used to evaluate the staining intensity. These results further validate the high expression of LEDGF/p75 in major human cancers.
Alcohol Affects the Late Differentiation of Progenitor B cell
Authors: Ewing Jacquelyn Bowers , Chan Wendy , Wolcott Michael , Wang Hao
Published: June 2021

Background

Studies showed in vitro alcohol exposure can affect the differentiation of progenitor B cell. This study investigates developmental stages on B cell differentiation and how alcohol affects these stages.

Methods

Oligoclonal‐neonatal‐progenitor (ONP) cells were cultured in vitro with/without the exposure of alcohol. Phenotype analysis was performed at various time‐points and the expression levels of transcription factors (TF) were also measured.

Results

ONP cell differentiated into B220‐CD11b− and B220‐CD11b+ cells in 3 days. No statistically significant difference of B cell formation was found in both normal and alcohol groups when exposed to alcohol during the first 3 days. If alcohol was added at day 4 and exposed to the end of culture, few yielded B cells. The expression level of TFs was down regulated with 100mM alcohol.

Conclusions

ONP cells can differentiate into lymphoid or myeloid lineage but diverged at B220‐CD11b−/B220‐CD11b+ branch point. Alcohol affects the ONP cell differentiation into B lineage at a late stage after cell divergence into B220‐CD11b−. Alcohol down regulates TFs expression resulting in poor B cell differentiation.
Activation of AP‐1 Signaling Pathway in Breast Epithelial Cells by Direct and Secondhand Cigarette Smoke
Authors: Mukherjee Shyamali , Das Salil K
Published: June 2021

AP‐1, a dimeric protein complex composed of Jun and Fos proteins, plays a key role in the control of cell death, proliferation and transformation. Several epidemiological studies have linked cigarette smoking to breast cancer, particularly in young women. Due to difficulties in conducting such studies in humans, we have developed and standardized a smoke exposure model system for human breast cell lines. This is a modified version of CULTEX cell culture smoke exposure system using the principle of the air/liquid exposure technique. Cells ( MCF10, MCF 12A and MCF 12F) were grown in culture media (DMEM/F‐12) and exposed to both direct and secondhand smoke generated from 1R3F cigarettes in a THRI MS‐SS smoke exposure systems. A sham control group was also used where cells were exposed to the same environmental condition without smoke exposure. Cells were isolated after 2 days and western blot analysis was performed to determine whether smoke exposure causes any change in the Jun and Fos family protein levels. Both direct and secondhand smoke exposure caused a significant activation of AP‐1 transcription factors associated with an increase in the protein level of Fos. Furthermore, an upregulation of cyclin D1 and PCNA was observed. Both direct and secondhand cigarette smoke exposure caused a significant activation of AP‐1 signaling pathway in all three breast cell lins. (Supported by FAMRI Clinical Innovator Award # 062415).
Activation of the Akt/mTOR pathway by EGF and its family growth factors is mediated by G‐protein αi1 and αi 3
Authors: Cao Cong , Marshall John , Jiang Meisheng , Chu Wen‐Ming
Published: June 2021

The Akt/mTOR pathway is essential for cell growth, migration and survival mediated by EGF and its family growth factors. However, the molecular mechanism underlying the activation of this pathway by EGF remains elusive. Here we report that G‐protein αi1 (Gi1) and Gi3 are critical for this activation process. Upon EGF stimulation, Gi1 and Gi3 associated with Gab1, which is required for activation of PI3K and Akt. Loss of Gi1/3 diminished EGF‐induced Gab1 phosphoryation and altered Gab1's association with PI3Kp85a. As a consequence, ablation of Gi1/3 severely impaired activation of Akt and mTOR's downstream events 4E‐BP1, S6K and S6 by EGF, HB‐EGF, and TGFα, but not by insulin, IGF and PDGF, leading to a defect in EGF‐induced cell growth, migration, cell survival and cyclinD1 expression. Thus, our findings define a novel EGF/Gαi/Gab1/PI3K/Akt/mTOR pathway and portray a genetic role for Gαi in the multiple cellular processes in response to EGF.
Integrin α11 mediates CCN1‐induced focal adhesion and β‐catenin translocation in esophageal epithelial cells
Authors: Modak Cristina , Mouazzen Wasim , Nguyen Anthony , Pham Jennifer , Chai Jianyuan
Published: June 2021

Background

CCN1 is a matricellular protein that plays a critical role in several normal and pathological processes by signaling through different integrin receptors in a cell‐type specific manner. Clinical studies have suggested CCN1 as a putative marker for malignant progression towards esophageal adenocarcinoma. However, little is known about its signaling mechanisms in this tissue. The aim of this study was to determine the integrin receptor that mediates CCN1 actions in esophageal epithelial cells.

Methods

Esophageal epithelial cells were treated with recombinant CCN1 protein. Its effects on integrins were examined by microarray, real‐time PCR, immunoprecipitation, immunoblotting, and immunofluorescence microscopy.

Results

Out of all the known α‐ and β‐integrins, ITGA11 and ITGB5 were the only ones that responded to CCN1 treatment. Co‐immunoprecipitation assays demonstrated an association between integrin α11 and β5 in the presence of CCN1. Blocking either integrin subunit prevented CCN1‐induced focal adhesion and β‐catenin nuclear translocation. Taken together, these results provide evidence that integrin α11 can form a new partnership with β5 and that together they serve as a new receptor to mediate CCN1 actions in esophageal epithelial cells.

This study was supported by the Department of Veterans Affairs of the United States and the American Heart Association.
Monomeric rhodopsin in nanodiscs is phosphorylated by rhodopsin kinase and binds arrestin with high affinity in the presence of negatively charged lipids
Authors: Vishnivetskiy Sergey A , Bayburt Timothy H , Morizumi Takefumi , Huang Chih‐chin , Tesmer John J , Ernst Oliver P , Sligar Stephen G , Gurevich Vsevolod V
Published: June 2021

The role of dimerization of class A GPCRs is hotly debated. We show that monomeric rhodopsin in nanodiscs is phosphorylated by rhodopsin kinase (GRK1) and binds arrestin1 as well as in native disc membranes. The efficiency of rhodopsin phosphorylation is the same in phosphatidylcholine (PC) nanodiscs containing 0–50% phosphatidylserine (PS). In contrast, arrestin1 binding to P‐Rh* in pure PC is low. Binding increases with the proportion of PS and plateaus at ~10‐fold higher level at ~30% PS. To investigate the role of PS in the arrestin1‐P‐Rh* interaction, we used structurally different “enhanced” arrestin1 mutants that bind P‐Rh* and Rh* with comparable affinity. Truncated arrestin1(1–378) that has no C‐tail and 3A mutant where the interaction anchoring the C‐tail to the body of the molecule is destabilized were significantly less dependent on PS, demonstrating >50% of maximum binding in pure PC. In contrast, polar core mutants with the same phenotype, R175E and D296R, depend on PS almost as much as WT arrestin1. Thus, monomeric receptor is normally phosphorylated and binds arrestin. Negatively charged lipids promote interaction by “pushing” the arrestin C‐tail away from the receptor‐binding surface. This is the first indication of the function of multiple negative charges present in the C‐tail of all arrestin subtypes. NIH grants EY011500 (VVG), GM033775 (SGS), HL071818 (JJT), and DFG Sfb740 and ER 294/1‐1 (OPE)
PACAP induces axonal elongation in cultured monkey trigeminal ganglion cells
Authors: Nakajima Emi , Walkup Ryan D. , Shearer Thomas R. , Azuma Mitsuyoshi
Published: June 2021

LASIK refractive surgery is widely accepted for correcting myopia. However, flap creation during LASIK surgery leads to complications including dry eye, caused by degeneration of sensory nerves. Our previous studies in rabbits showed that PACAP accelerated the extension of neuronal processes and recovered corneal sensitivity. The purposes of the present study were to 1) establish a cell culture system for monkey trigeminal nerve (TgN), whose fibers innervate cornea, and 2) determine if PACAP induces sprouting and elongation of axons. Expression of mRNA and cellular distribution for the PACAP receptor, PAC1, were determined by qPCR and immunostaining. Axonal length in cultured TgN ganglion cells was evaluated by staining with antibody for neurofilament. Our results showed that dissociated, cultured TgN ganglion cells contained neuronal and Schwann cells. The bioactive N‐terminal peptide PACAP27 significantly induced axonal elongation which was inhibited by the PAC1 antagonist PACAP 6–27. Downstream inhibitors of the PAC1 signaling pathway, AC and PKC, also inhibited axonal elongation. Together with positive PAC1 staining in neuronal cells, our data suggested involvement of PAC1 activation in axonal elongation. PACAP might be a candidate drug for treatment of complications after LASIK.

Dr. Shearer receives consulting fees from, and Drs. Azuma and Nakajima are employees of, Senju Pharmaceutical Co. Ltd.
Neuroprotection by estrogen under hypoglycemic condition with special emphasis on AKT_GSK pathway
Authors: Chakraborty Sanjoy , Simeon Stephanie , Palmieri Vincent , Addya Sankar
Published: June 2021

Our previous studies have shown that estrogen is involved in the neuroprotection of cells under hypoglycemic conditions within 24 hrs. The hypothalamic cells (N38) were treated under various conditions (T1‐ media with 5%FBS and 1% Pen‐Strep at 24 hrs, T2‐ T1without glucose at 24 hrs, T3‐ T1 without glucose with 100 micro M estrogen, T4‐ T1 at 72 hrs, T5‐ T2 at 72hrs, T6‐ T3 at 72 hrs.). One of the possible pathway through which estrogen exerts its protection is through Akt‐GSK coupling. Akt regulates cell survival by phosphorylating components of cell death apparatus like glycogen synthase kinase‐3 (GSK3b). When the activity of AKT is at its highest, GSK3b is in its inhibited state leading to cell survival and vice versa. DNA microarray studies showed that AKT was down‐regulated and glycogen synthase upregulated in the absence of glucose and in the presence of estrogen there was an increase and decrease respectively in gene expression. Network/functional analysis gave further information of other signaling molecules upregulated by the estrogen treatment after hypoglycemic shock like MMP3, POSTN, LOX, PENK, TGFB1 at 72 hours whereas within 24 hours neurotrophin, NGF, KLK2, IF144 were upregulated and VMA21 downregulated. There were 11 genes that showed 1.5 fold change and 1 gene showing 2 fold change. Between T5 vs. T6 there were 58 genes that showed 1.5 and 3 genes showed above 2 fold change. The study raises the possibility of novel pharmacological interventions against hypoglycemia and use of estrogen against the neuronal deficits. The result supports ovarian steroid as modulators in CNS responsiveness to recurring glucoprivation and its mechanism of action through membrane receptors.
Targeting a Prokaryotic Protein in a Eukaryotic Parasite
Authors: Hedstrom Lizbeth
Published: June 2021

Cryptosporidium is a major cause of the “vicious cycle of diarrhea and malnutrition”, an important AIDS pathogen and potential bio‐terrorism agent. The currently approved drugs to treat cryptosporidiosis are ineffective, and no vaccines exist. Genomic analysis indicates that inosine monophosphate dehydrogenase (IMPDH) is required for the production of guanine nucleotides. Moreover, the parasite obtained its IMPDH gene from bacteria by lateral gene transfer, so that this enzyme is very different from the host counterpart. We designed a high throughput screen to target the most diverged portion of the IMPDH active site. This screen has identified fifty parasite‐selective IMPDH inhibitors. These compounds bind in the NAD site as expected. The x‐ray crystal structure of the inhibitor complex reveals the structural basis for selectivity as well as the spectrum of inhibition of IMPDHs from bacterial pathogens. We have also developed a novel assay for antiparasitic activity by re‐programming the guanine biosynthetic pathways of Toxoplasma gondii to mimic Cryptosporidium. Medicinal chemistry optimization has produced inhibitors with nanomolar affinities. The best inhibitors display antiparasitic activity with greater potency than paromomycin, the current “gold standard” for anticryptosporidial activity. Supported by NIH RO1 AI055268 (Boris Striepen, UGA) and NIH U01 AI075466 (L.H.).
MORPHINE MODULATION OF THROMBSPONDIN LEVELS IN ASTROCYTES AND ITS IMPLICATIONS FOR SYNAPSE FORMATION
Authors: Coscia Carmine James , Ikeda Hiroko , Miyatake Mayumi , Koshikawa Noriaki , Ochiai Kuniyasu , Yamada Kiyoshi , Kiss Alexi , Donlin Maureen , Panneton W. Michael , Churchill James , Green Michael , Siddiqui Akbar , Leinweber Andrew , Ezerskiy Luba , Rendell Victoria , Belcheva Mariana
Published: June 2021

Opioid receptor signaling via the ERK/MAP kinase phosphorylation cascade initiates diverse cellular responses that are cell type dependent. In astrocytes, multiple μ opioid receptor mediated mechanisms of ERK activation exist that are temporally distinctive and feature different outcomes. Upon discovering that chronic opiate treatment of rats followed by spontaneous withdrawal down regulates thrombospondin1 (TSP1) expression in the nucleus accumbens and cortex, we investigated the mechanism of action of this modulation in astrocytes. TSP1 is synthesized in astrocytes and released into the extracellular matrix where it plays a role in synapse formation. Acute morphine (h) reduced TSP1 protein levels in astrocytes by an ERK dependent mechanism. Chronic (days) opiates repressed expression of TSP1 and reduced its protein levels. Morphine reduced TSP1 levels stimulated by TGFβ1. Accordingly, chronic morphine treatment of astrocyte neuron co‐cultures reduced neurite outgrowth and synapse formation. Taken together our results suggest that inhibitory actions of morphine on TSP1 and synapse formation entail multiple temporally distinctive mechanisms. Supported by DA005412.
Resistance to cardiac doxorubicin damage in a Gsta4 knockout mouse
Authors: Benes Helen , Vuong Mai , Telemaque Sabine , Marsh James D , Zimniak Piotr , Boerma Marjan
Published: June 2021

4‐hydroxynonenal (4‐HNE) mediates both pathological effects of oxidative stress and signaling from mitochondrion to nucleus to elicit transcriptional responses that are largely controlled by Nrf2. As elevated levels of 4‐HNE are associated with several cardiovascular diseases, metabolism of 4‐HNE is critical. The murine glutathione transferase A4–4 (GSTA4–4) exhibits very high levels of 4‐HNE‐conjugating activity, which is reduced in the hearts of Gsta4 knockout (KO) mice. KO mice exhibited no cardiac pathology; cardiac 4‐HNE levels were no different in WT and KO mice. We examined how Gsta4 KO mice react to high cardiac 4‐HNE accumulation following chronic administration of doxorubicin (DOX) (5 mg/kg, 4 weekly injections). Cardiac function was assessed by echocardiography. While untreated mice exhibited similar basal cardiac function, WT mice were more sensitive to DOX, resulting in a significantly higher mortality rate. Ejection Fraction (EF) fell in both groups after the first DOX injection. At day 21, EF was significantly lower in WT mice as compared to baseline whereas EF in KO mice recovered to the basal value. Changes in fractional shortening correlated with EF in both groups. We concluded that Gsta4 KO mice exhibited resistance to DOX cardiomyopathy and will explore how Nrf2‐induction of SOD and catalase may contribute to a cardioprotective effect. Supported by a College of Medicine Pilot Study Grant.
Activation of DNA topoisomerase I and DNA topoisomerase II in nuclei of interleukin‐2‐treated HuT 78 cells
Authors: Foglesong Paul David , Pham Minh‐Hieu , Kondapalli Navya , Singh Thakur Aditya , Jaiswal Tripti , Ma Jingjing , Contreras Leonard
Published: June 2021

The activities of DNA topoisomerase I and DNA topoisomerase II were measured in HuT 78 cells, a human T cell lymphoma cell line, after treatment of the cells with interleukin‐2 (IL‐2). HuT 78 cells were treated with 1,000 units IL‐2/ml, and extracts of the nuclei were prepared from 0 to 12 hours after treatment. The activity of DNA topoisomerase I was assayed by relaxation of supercoiled pBR322 DNA. The activity of DNA topoisomerase II was assayed by unknotting of P4 DNA, decatenation of kinetoplast DNA, and cleavable complex formation. The decatenation activity was sensitive to etoposide, an inhibitor of DNA topoisomerase II. The specific activities of both enzymes were determined in units/mg protein. The specific activities of both DNA topoisomerase I and DNA topoisomerase II increased 3‐ to 11‐fold in nuclear extracts prepared from HuT 78 cells in three transient, concomitant peaks observed at 0.5, 4, and 10 hrs after treatment with IL‐2. The specific activities of both enzymes returned to baseline values after each peak. These results indicate that the activities of DNA topoisomerase I and DNA topoisomerase II are extensively regulated in activated human T cells, and they suggest that both DNA topoisomerase I and DNA topoiosmerase II function in both transcription and DNA replication in IL‐2‐activated T cells. This work was supported by grants from the Morrison Trust and the NIH (HD 52388).
The Response of the ERK/MAPK Cascade to Acute Stress in the Medial Prefrontal Cortex of Chronically Stressed Rats
Authors: Lam Mai Truc , Ferland C. L. , Schrader L. A.
Published: June 2021

The chronic variable stress (CVS) rodent model mimics the effects of chronic stress in humans and provides an animal model of stress‐induced depression. The medial prefrontal cortex (mPFC) is believed to negatively regulate the hypothalamo‐pituitary‐adrenocortical axis (HPA), a major system involved in stress response, through the release of glucocorticoids. The function of the mPFC may be dysregulated in chronic stress, mediated by cellular signaling cascades. Studying signaling mechanism changes in the mPFC is critical to improving our understanding of stress‐induced disorders. This study investigates acute stress‐mediated activation of the extracellular signal‐regulated kinase (ERK) cascade and the transcription factor cAMP response element binding (CREB) in the mPFC under control and CVS conditions through Western Blotting. Preliminary data show that CVS animals have reduced levels of ERK activation in the mPFC compared to control animals. Acute stress may act on CVS and control groups differently suggesting a differential response of the ERK/MAPK cascade to acute stress in the mPFC. This may provide a potential mechanism for dysregulation of the HPA axis under chronically stressful conditions.
Kruppel‐like factor 4 is involved in TGF‐beta1‐induced VSMC differentiation through trans‐activating TbetaRI expression
Authors: Li Huixuan , Han Mei , Zheng Bin , Sun Shaoguang , Su Ming , Fu Jian‐ran , Wen Jin‐kun
Published: June 2021

Krüppel‐like factor 4 (KLF4) is involved in phenotypic switching of vascular smooth muscle cells (VSMCs) induced by transforming growth factor beta (TGF‐β). However, as a non‐Smad protein, the role of KLF4 in TGF‐β‐regulated biological processes remains unclear. In this study, we found that TGF‐β1 inhibited cell cycle progression and induced expression of differentiation marker genes in cultured VSMCs, which was associated with the up‐regulation KLF4 and TGF‐β type I receptor (TβRI) expression induced by TGF‐β1. TGF‐β1 induced KLF4 phosphorylation in VSMCs, Which was blocked by pretreating cells with either SB431542, the TGF‐β inhibitor, or SB203580, the p38 MAP kinase inhibitor. We further demonstrated that treating VSMCs stimulated KLF4 interaction with Smad2, indicating that the phosphorylation of KLF4 is required for the interaction of KLF4 with Smad2 and KLF4 mediated TβRI expression via activation Smad and p38 MAP kinase signaling. In addition, TGF‐β1 increased DNA binding activity of KLF4 and Smad2 to TβRI promoter. We demonstrated that two potential KLF4 binding sites could activate TβRI promoter, and meanwhile, KLF4 formed complex with Smad2 to bind to the Smad‐binding region and activate the TβRI transcription cooperatively. These results indicate that KLF4 plays a positive regulatory effect on cell differentiation through trans‐activating TβRI expression via synergistic interaction with Smad2 in TGF‐β‐induced VSMCs.
Dynamic cross‐talk between PI3‐kinase/Akt and Ras/ERK pathways in EGF receptor signaling that can affect drug sensitivity in tumor cells
Authors: Aksamitiene Edita , Kiyatkin Anatoly , Hoek Jan B
Published: June 2021

Cell survival (PI3K/Akt) and mitogenic (Ras/ERK) signaling cascades rarely act as independent parallel pathways, rather they interact at different points and phases of signal propagation, generating positive and negative feedback loops. We explored the patterns of cross‐talk between PI3K and ERK in various normal and cancer cells that were pretreated with wortmannin or U0126 inhibitors prior to stimulation with increasing EGF doses. Time‐courses of total and phosphorylated ERK or Akt expression were detected by Multistrip Western blotting, a novel procedure used for concurrent comparison of signals derived from multiple blots. Quantitative analysis of blots showed that PI3K enhanced and sustained ERK responses, whereas activated ERK suppressed PI3K activity in time‐ and EGF dose‐dependent manner. The differential extent of such bidirectional reciprocal crosstalk could be attributed to diverse recruitment mechanisms of GAB family adaptor proteins, as suggested by immunoblot analyses of subcellular fractions and immunoprecipitates. Dynamic PI3K‐ERK interactions also persist in tumors, making the activation patterns robust to individual perturbations, which can be linked to different drug sensitivity profiles. Nonetheless, combined inhibition of PI3K and ERK activities synergistically decreased tumor cell viability and growth, as measured by AlamarBlue assay. Supported by NIH Grant GM059570
Exosome‐release of beta‐catenin: A novel mechanism to antagonize Wnt signaling
Authors: Chairoungdua Arthit , Smith Danielle , Pochard Pierre , Hull Michael , Caplan Michael
Published: June 2021

Expression of the tetraspanins CD82 and CD9 suppresses tumor metastasis. Previous studies have shown that CD82 and CD9 are enriched in exosomes. The functional roles of tetraspanins in exosomes and their roles in the suppression of tumor cell metastasis remain unknown. We find that CD82 and CD9 serve as negative regulators of the Wnt/β‐catenin signaling pathway, whose inappropriate activation is involved in the development and progression of several cancers. CD82 and CD9 expression strongly suppressed Wnt/β‐catenin activity as measured by luciferase reporter assay and induced a significant decrease in β‐catenin protein levels in both cytoslic and nuclear fractions in HEK 293T cells. In addition, RNAi knock down of endogenous CD9 resulted in an increase in β‐catenin levels and Wnt/β‐catenin activity. CD9 and CD82 expression inhibited Wnt/β‐catenin signaling through a mechanism independent of GSK‐3β and of the proteasome and lysosome‐mediated protein degradation pathways. CD82 and CD9 expression enhanced β‐catenin transport into exosomes via a mechanism that was blocked by sphingomyelinase inhibitor, GW4869. CD82 failed to induce exosome release of β‐catenin in CHO cells, which do not express endogenous E‐cadherin. However, β‐catenin and E‐cadherin were detected in exosomes purified from the culture medium of CHO cells that were cotransfected with CD82 and E‐cadherin. Exosome release from dendritic cells generated from CD9 knockout mice was reduced compared to dendritic cells generated from wild‐type mice. Taken together, our results indicate that CD82 and CD9 might suppress tumor metastasis through downregulation of the Wnt signaling pathway via a novel mechanism that leads ultimately to the exosomal discharge of β‐catenin.
Effect of SEB induced cell death in DU 145 cells
Authors: Zindel Kristin , Wiewel Kurstin , Hammamieh Rasha , Jett Marti , Mendis Chanaka
Published: June 2021

Staphylococcal enterotoxin B (SEB) is one of the enterotoxins that have been thoroughly investigated in multiple cell lines; however, little is known about the cascades of signaling events that explain its patho‐mechanism. This study follows a molecular route to better understand the apoptosis related events known to be induced by SEB in a cancer cell line, DU 145. DU 145 is a prostate cancer cell line which has moderate metastatic potential. As SEB is known to induce multiple signal transduction pathways, we have identified a crucial pathway inter‐connector (JNK) that may have an inhibitory effect in SEB induced unwanted cellular activities including cell death. Our study was centered on inhibiting pathway inter‐connector JNK by using SP600125 in this cancer cell line, followed by an evaluation of the efficacy of the target through alterations to a set of known apoptosis and cell growth related genes. We have quantified the RNA and protein samples, and have carried out gene expression analysis. As our experimental design specifically targeted pathway inter‐connector JNK, we believe that the target is unique and has the ability to sustain longer lasting inhibitory effects.
Effect of helicobacter pylori infection on iron absorption in asymptomatic adults consuming iron and zinc fortified wheat
Authors: de Romaña Daniel Lopez , Diazgranados Doricela , Pizarro Fernando , Ruz Manuel , Brunser Oscar
Published: June 2021

Introduction

Micronutrient deficiencies and Helicobacter pylori (HP) infection are prevalent and co‐exist in several countries.

Objective

The objective of the study was to determine the effect of HP on iron absorption in asymptomatic adults consuming iron and zinc fortified wheat.

Materials/Methods

HP was assessed using the C13‐urea breath test. On day 1 subjects received for breakfast bread fortified, per Kg flour, with either 55 mg ferrous sulfate and 60 mg zinc sulfate, labeled with 59Fe, as sulfate, or same amounts of ferrous fumarate and zinc oxide, labeled with 55Fe, as fumarate. On day 3 they received the other type of bread. On day 17 blood (20 ml) was drawn to assess circulating radioactivity. Iron absorption was determined using Eakins and Brown's double isotopic technique.

Results/Findings

Prevalence of HP was 78%. Fourteen HP negative and 16 HP positive individuals completed absorption studies. There were no significant differences in the mean iron absorption of either ferrous sulfate or ferrous fumarate by infection status.

Conclusion

HP has no effect on iron absorption in asymptomatic adults consuming iron and zinc fortified wheat.

Support by FONDECYT grant 1080032
Negative Iron Balance: Risk Among Female Vegetarian College Students
Authors: Hawk Susan N. , Englehardt Kimberly G.
Published: June 2021

Iron deficiency anemia is the most common nutrient deficiency, impacting up to 6% of Americans. Given that 5 to 14% of women in the United States between ages 15 and 44 have impaired iron status, depletion may be most prevalent in younger women. Of particular concern is the health impact of iron deficiency anemia among vegetarian women of child bearing age. The objective of this study was to determine if iron deficiency anemia is widespread among college age vegetarian women. A cross sectional analysis of 39 female students at California Polytechnic State University in San Luis Obispo, CA was conducted. Anthropometric data including height, weight and BMI was collected. Three day dietary records were maintained by participants to determine dietary iron intake. A fasting blood sample was obtained and analyzed for white blood cells, red blood cells, hemoglobin, hematocrit, serum iron, serum ferritin, transferrin saturation and total iron binding capacity. Vegetarians tended to eat less iron daily than non vegetarians. Of interest was the tendency for lower serum ferritin status and a greater expression of stage 1V negative iron balance compared to non vegetarians as evidenced by low serum iron, high total iron binding capacity, low transferrin saturation and low serum ferritin. Thus, vegetarian college age women appear to be at higher risk than non vegetarians for developing negative iron balance. Funded by California Polytechnic State University Faculty Support Grant.
Hemoglobin regeneration efficiency method revisited: an alternative bioassay for iron bioavailability
Authors: Yokoi Katsuhiko , Konomi Aki , Otagi Miki
Published: June 2021

Iron (Fe) deficiency is a worldwide problem, and a search for foods containing bioavailable Fe is an effective measure. The slope‐ratio method using graded levels of dietary Fe vs hemoglobin (Hb) are commonly used for the estimation of the bioavailability of Fe in foods, but it requires a considerable number of rats and do not give a value for individuals. The objective was a comparison between the Hb regeneration efficiency (HRE) method and the slope‐ratio method.

HRE method

Twenty‐one F344/N male weanling rats were fed a Fe‐deprived diet for 4 weeks to deplete body Fe stores. Then, three groups of seven rats each were repleted for 20 days using diets fortified with ferrous sulfate, ferric citrate, or cocoa powder (van Houten brand) each provided 20 mg Fe/kg diet. The diets were based on the AIN‐93G formula. HREs were 0.733, 0.350 and 0.336 for ferrous sulfate, ferric citrate and cocoa powder, respectively. The HRE of Fe in cocoa powder was 96% that of ferric citrate.

Slope‐ratio method

Another 32 weanling rats were depleted in Fe stores for 4 weeks and then 8 groups of four rats each were repleted for 25 days using graded levels of dietary ferric citrate or cocoa powder. The slope for cocoa powder was 99% that of ferric citrate.

In summary, the HRE method gave a similar outcome to the slope‐ratio method. The bioavailability of Fe in cocoa was significantly less than that of ferrous sulfate and was similar to that of ferric citrate.
Relation of infant iron status at 12 and 18 months to neonatal iron status and postnatal iron intake
Authors: Wachs Theodore D. , Creed‐Kanashiro Hilary
Published: June 2021

Does neonatal iron status predict infant iron status at 12 and 18 months, over and above the contributions of postnatal dietary iron intake? At 12 months our sample consisted of 241 infants from low income families living in Lima Peru. Because of funding issues we could only follow‐up 104 of these infants to 18 months. Our data base consisted of maternal hemoglobin assessed twice during pregnancy, neonatal serum ferritin, transferrin saturation and hemoglobin taken from cord blood, 6,12 and 18 month iron intake taken from 24 hour dietary recalls and 12 and 18 month transferrin saturation and hemoglobin levels taken from venous blood. Models testing the prediction of 12 or 18 month transferrin saturation were non‐significant. 12 month hemoglobin was predicted (R2 = .12, p<.01) by neonatal ferritin (Beta = .24, p<.01),, neonatal hemoglobin (Beta, .18, p<.05) and 12 month dietary iron intake (Beta = .18, p<.05). The model remained significant after covarying out family demographics and non‐iron dietary intake. 18 month hemoglobin was predicted (R2 = .44, p<.01) only by 12 month hemoglobin (Beta = .63, p<.01) and remained significant when covariates were entered. The overall pattern suggests that both neonatal iron status and dietary iron intake uniquely predict 12 month iron status, which in turn is a primary predictor of later hemoglobin. Research supported by the National Science Foundation.
Effect of dietary iron deficiency and overload on hepatic ZIP transporter expression in rats
Authors: Nam Hyeyoung , Knutson Mitchell D
Published: June 2021

The mammalian ZIP (Zrt‐, Irt‐like Protein) family of transmembrane transporters consists of 14 members. Although ZIPs have been characterized largely by their ability to transport zinc, some ZIPs can transport other essential metal ions as well, including Fe2+ and Mn2+. We sought to determine if iron deficiency or overload affected the expression of ZIP transporters in the liver, a main site of iron storage. Weanling male rats (n=6/group) were fed diets containing either 9 ppm Fe (deficient), 215 ppm Fe (control), or 2.8% carbonyl Fe (loaded). After 3 wk, liver non‐heme iron levels were 15, 87, and 5223 μg/g. Q‐RT‐PCR analysis of ZIP mRNA levels identified an 8‐fold induction in ZIP5 mRNA levels in iron‐loaded livers, and reciprocal regulation of ZIP10 by iron (twice as high in deficiency as in overload). The iron‐related changes in ZIP5 and ZIP10 expression were confirmed in a second study of rats fed AIN93G diets providing 10 ppm Fe, 50 ppm Fe, or 1.9 % carbonyl Fe, resulting in liver non‐heme Fe concentrations of 13, 30, and 1813 μg/g. These studies suggest that ZIP5 and ZIP10 may play a role in hepatic iron/metal homeostasis during iron deficiency and iron overload.
Association of cognitive impairment and zinc intake in HIV+ adults
Authors: Mandal Priyanka , Baum Marianna K , Sales Sabrina , Li Yinghui , Jayaweera Dushyantha T , Page John B , Campa Adriana
Published: June 2021

Background

The central nervous system contains 10% of all body zinc. Zinc deficiency, which is prevalent in HIV‐infection, has been associated with cognitive impairment. This study examined the risk factors associated with impaired cognitive performance in HIV infection.

Methods

After consenting 231 HIV‐infected adults, a 30 point‐questionnaire assessed cognitive function by testing immediate memory, problem solving ability and quality of life. Those with <26 points were considered cognitively impaired. Blood was drawn for plasma levels of zinc and HIV disease parameters. Quality of life, alcohol and drug use, and 24‐hour dietary recall questionnaires were administered.

Results

There were 54 cases (23.3%) with cognitive impairment. When compared to those without impairment, cases had significantly poorer dietary zinc intake (8.1±SD5.7 vs. 10.4±SD7.8 mg, p=0.027) and were significantly older (44.8±SD6.3 vs. 42±SD7.1 years, p=0.0088). Alcohol and illicit drug use and frequency of use, plasma levels of zinc, CD4 count<200 cell/mm3, gender, and quality of life scores were not significantly different between groups.

Conclusion

Lower zinc intake is a risk factor for impaired cognitive performance in this cohort of HIV+ adults. Prospective studies are needed to examine whether zinc supplementation might alleviate cognitive impairment in HIV+ adults with zinc deficiency.

Funded by NIDA
Lycopene suppressed leptin‐mediated malignant progression in human colon cancer cells
Authors: Lin Meng‐Chiu , Tang Feng‐Yao
Published: June 2021

Colorectal cancer is one of the leading causes of cancer death in western countries. Previous studies indicate that leptin induces cellular proliferation through multiple signalling pathways in colon cancer cells. Recent studies suggested that loss of E‐cadherin adherent molecule has been associated with epithelial–mesenchymal transition (EMT) and the malignant progression of human colon cancer cells. Our previous studies indicated that lycopene could exhibit anti‐cancer effects on human colon cancer cells. However, the inhibitory effect of lycopene on leptin‐mediated malignant progression of human colon cancer has not been demonstrated well yet. The present study demonstrated that leptin dose‐dependently (0, 0.0125, 0.0625, 0.25μM) induced cellular migration and invasion through up‐regulation of matrix metalloproteinase (MMP)‐7 as well as suppression of E‐cadherin in human colon cancer HT‐29 cells. The molecular mechanisms of actions were through the modulation of MAPK/JNK and JAK/STAT‐3 signaling pathways. Furthermore, our results demonstrated that lycopene (2 μM) effectively inhibited leptin‐mediated cellular migration through up‐regulation of E‐cadherin molecule and modulation of JNK signaling pathway in human colon cancer HT‐29 cells. Take together; our results suggest that lycopene could act as an effective chemo‐preventive agent against the leptin‐ mediated malignant progression in human colon cancer cells.

Grant Funding Source: National Science Council grant under agreement No. 97‐2320‐B‐039‐043‐MY3
Dietary inhibitors of monoamine oxidase A: quercetin could be a naturally‐occurring antidepressant
Authors: Ramsay Rona Ruth , Dixon Clarke Sarah E.
Published: June 2021

Health supplements are frequently sought to alleviate low mood. The pharmacokinetics of components of tobacco and on flavonoids were examined to see whether a normal diet provides compounds active as monoamine oxidase inhibitors in sufficient quantities to affect mood. A literature search identified dietary monoamine oxidase inhibitors, their levels in foods, the pharmacokinetics of absorption, and the distribution and tissue levels observed. Estimated daily intake and expected tissue concentrations were compared with the efficacies of the compounds as inhibitors of monoamine oxidases from the literature and, for monoamine oxidase A, assayed spectrophotometrically.

Norharman, harman and quercetin can be present in a healthy diet at up to 0.15 mg norharman, 0.11 mg harman and 54.2 mg quercetin per day. The pharmacokinetics and tissue levels for these compounds were consistent with significant levels reaching neuronal monoamine oxidase from the diet or smoking. Although diverse methods in the pharmacokinetic studies make the comparisons qualitative, quercetin was the highest in the sample diet and was competitively inhibited monoamine oxidase A. Quercetin in the diet could be a natural antidepressant compound.

Supported by a Biotechnology and Biological Sciences Research Council Vacation Bursary.
Anti‐cancer effect of Korean radish(Raphanus sativus L.) leaf extract through induction of apoptosis and down‐regulate Erb B2 signaling and Akt pathway in human breast MDA‐MB‐231 cell lines
Authors: Chang Moon‐Jeong , Kim Boram , Lee Hye‐Sung , Kim Woo‐Kyoung , Cho Kangjin
Published: June 2021

The vegetative portion of radish have not been characterized in terms of its biological activity. In vitro anti‐cancer effect of aqueous extract from Korean radish leaf (EKRL) was evaluated in human breast MDA‐MB‐231 cell lines. EKRL preferentially inhibited the growth of cell lines in a dose‐dependent manner (p<0.05). In addition, EKRL treated MDA‐MB‐231 decreased expression of anti‐apoptotic Bcl‐2 protein and unchanged expression of pro‐apoptotic Bax protein, eventually leading a decrease in the Bcl‐2/Bax ratio. Phosphorylated Akt levels markedly reduced in cells treated with 100ug/L EKRL and total Akt levels decreased at 300ug/L. Furthermore, a dose‐dependent decrease in Erb B2 and Erb B3 levels was detected in EKRL treated cells. These results demonstrated that EKRL inhibits the proliferation of MDA‐MB231 through induction of apoptosis and down‐regulate Erb B2 signaling and the Akt pathway, making EKRL a potent candidate as new‐anti cancer food components.

(This research was supported by Korea Rural Development Adminstration, project #2009AFT143783066)
Effects of Phellinus ribis Water Extracts(PRWE) on Apoptosis‐Induction in Liver Cancer Cell(Hepa1c1c7 cell)
Authors: Jeong Boyoung , Kim Dongyeop , Kim Gyoung Woo , Fan Jiang Ping , Han Gi Dong
Published: June 2021

The Phellinus ribis has been known to exhibit potent biological activities, such as anti‐viral, anti‐cancer and anti‐diabetes activities. This study investigated the effects of apoptosis induction in Hepa1c1c7 mouse liver cancer cells by treatment of PRWE. There were no cytotoxicity of PRWE evaluated by MTT assay with NCTC1469 normal cells and Hepa1c1c7 mouse cells. The changes of mRNA and proteins pivotally related to apoptosis were investigated using RT‐PCR and Western blots. The mRNA and protein expressions of TNF‐α, Caspase‐8 and Fas associated with death ligand/receptor‐dependent apoptosis pathway were increased in PRWE treatment group. The treatment of PRWE regulated mitochondria‐dependent apoptosis pathway by inducing the increase of Caspase‐9, Caspase‐3 and the decrease of Bcl‐2 in Hepa1c1c7 cells. In addition, the expression of p53 was increased in PRWE treatment group, indicating a direct damage on the cells. These results showed that PRWE effectively induced death of liver cancer cell by activating apoptosis.
Lipid components of 10 different nut types
Authors: Kornsteiner‐Krenn Margit , Elmadfa Ibrahim
Published: June 2021

The objective of this study was to determine beneficial constituents, which are responsible for the lipid‐lowering effects of nuts.

Almonds, Brazil nuts, cashews, hazelnuts, macadamia, peanuts, pecans, pines, pistachios and walnuts were investigated in terms of total oil content, fatty acid pattern, and phytosterol content. Total lipid content and fatty acid pattern were analysed with Soxhlet extraction and GC. The phytosterol content was determined with RP‐HPLC.

The fat content varies between 45% (cashews) and 75% (macadamias). Almonds, cashews, hazelnuts, macadamias and pistachios are good sources of monounsaturated fatty acids (> 55%). Peanuts and pecans are rich in monounsaturated and polyunsaturated fatty acids (both > 40%), whereas Brazil nuts, pines and walnuts have the highest polyunsaturated fatty acid content (> 50 %). The mean total sterol composition (including campesterol, ergosterol, stigmasterol and β‐sitosterol) oscillated from 48 mg (Brazil nuts) to 139 mg (Pistachios) per 100 g nuts. Campesterol, ergosterol and stigmasterol were minor phytosterol components (< 10 mg).

The results demonstrated that nuts are rich in unsaturated fatty acids and phytosterols, which are recognized for lipid‐lowering effects.
Effect of Sasa borealis bamboo Diet on Lipid Levels, Platelet Aggregation and Erythrocyte Na+ Efflux in OLETF Rats
Authors: Kang Ja Young , Lee Dae‐Ho , Kang Young‐Hee , Kang Jung Sook
Published: June 2021

This study was conducted to investigate the effects of Sasa borealis bamboo leaves on plasma and liver lipids, platelet aggregation and erythrocyte Na efflux using OLETF rats. OLETF and the normal counterpart LETO rats were divided two groups each and fed either control diet or diet with 10% Sasa borealis bamboo leaves for four weeks.

The final weight with high FER were significantly increased in OLETF rats (p<0.05). Plasma and liver total cholesterol and TG were significantly increased in OLETF rats compared with LETO rats (all, p<0.05). Plasma total cholesterol and TG in OLETF‐bamboo was significantly decreased compared with OLETF‐control (p<0.05, p<0.01). HDL‐cholesterol was significantly lower and blood glucose was significantly higher in OLETF than LETO rats (p<0.05, respectively). Platelet aggregation in initial slope was significantly decreased in OLETF rat (p<0.05), and bamboo reversed it. Intracellular Na was significantly increased in OLETF rats and bamboo diet significantly decreased (p<0.05). Na‐K ATPase and Na‐K cotransport tended to be lower and Na passive transport be higher in OLETF than LETO rats, and bamboo diet significantly decreased Na‐K cotransport in LETO and OLETF rats (p<0.05). Liver TBARS production decreased significantly in OLETF fed bamboo compared with OLETF‐control (p<0.05).

In present study, OLETF rats, type II DM model showed unfavorable parameters in terms of weight gain and blood and liver lipids, and hypolipidemic and antioxidant effects of Sasa borealis bamboo were more evident in OLETF rats.
SIGNIFICANT DECREASE IN HOMOCYSTEINE AND ox‐LDL IN HEALTHY HEAVY SMOKERS AFTER NUTRACEUTICAL SUPPLEMENTATION
Authors: Bamonti Fabrizia , Novembrino Cristina , Vigna Luisella , De Giuseppe Rachele , de Liso Federica , Pellegatta Marco , Gregori Dario , Della Noce Cinzia , Accinni Roberto
Published: June 2021

Nutraceuticals may attenuate some negative effects of cigarette smoking. We evaluated effects of two different encapsulated formulas, consisting primarily of mixed juice powder concentrate (Juice Plus+®), in a randomized placebo controlled, double‐blind study on 101 heavy smokers (>20 cigarettes/d, duration ≥10y, median age 47; 41–57 y, 63 M, mean capsule compliance 85.2%). Smokers were divided into 3 groups: A‐placebo, B‐fruit/vegetable, C‐fruit/vegetable/berry. Serum and erythrocyte folate (s‐fol and ery‐fol, respectively) and total homocysteine (tHcy) concentrations were measured before and after 3‐m supplementation. In 75 (46 M) compliant (>95%) subjects, lipid panel parameters (total Cholesterol, tChol; LDL‐c; oxidized‐LDL, ox‐LDL) were evaluated. Median values were compared (t‐test for paired data); data are reported as mean delta values (± standard deviation). Intervention with both nutraceutical formulations resulted in reduction in ox‐LDL and tHcy compared to placebo.

Support: NSA, Collierville, TN

A (placebo)
B (fruit/veg)
C (fruit/veg/berry)

tChol (mg/dL)
9.3 (±18.4)
−11.6 (±22.9)*
−6.3 (±25.5)*

LDL‐c (mg/dL)
16.8 (±17.6)
−8.1 (±17.9)*
−0.8 (±20.5)*

ox‐LDL (U/L)
−2.3 (±11.3)
−13.8 (±12.5)*
−11.2 (±12.2)*

tHcy (μM)
−0.14 (±1.4)
−2.38 (±2.3)#
−3.20 (±5.2)#

s‐fol (nM)
2.8 (±5.5)
13.7 (±9.9)#
16.6 (±9)#

ery‐fol (nM)
194 (±178.3)
488.2 (±349)#
646.3 (±362.1)#