bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression of TLR9 in mouse and human lung and its role in lung inflammation in chicken barn air exposure
Authors: Schneberger David , Aulakh Gurpreet , Channabasappa Shankaramurthy , Caldwell Sarah , Singh Baljit
Published: June 2021

Workers exposed to intensive animal housing facility air show signs of occupationally‐related lung dysfunction. These respiratory symptoms are associated with endotoxin, but within these environments gram negative bacteria may constitute only a small portion of the microorganisms present. In contrast, unmethylated DNA can be found in all bacteria, some viruses, and mould. Immune responses to this unmethylated DNA are mediated through the TLR9 receptor. Therefore, we sought to test expression of TLR9 in mouse as well as human lungs, to see if mice would be an appropriate model for an exposure study. We hypothesized that the immune response seen in the lungs of mice exposed to barn air would be altered in a TLR9‐deficient animal towards a less inflammatory state.

We used immunohistology, immuno‐electron microscopy and in situ hybridization to show expression of TLR9 on bronchial epithelium, alveolar septal cells, and alveolar macrophage of mouse and human lungs. Having established similar receptor expression, a barn exposure study was done using a TLR9‐deficient mouse model. Mice were exposed to chicken barn air for 8 hours/day for 1, 5, or 20 days. Examination of bronchiolar lavage and serum against a panel of cytokines (IL‐1β, IL‐6, IL‐10, IL‐12, TNF‐α, and IFN‐γ) showed no significant differences after one day exposure. TNF‐α (p=0.06) levels in TLR9‐deficient mice were reduced in blood and lavage fluids after 5 days, and somewhat reduced at 20 days of exposure (p=0.14), while IFN‐γ was also reduced at 5 days(p=0.06) and remained reduced after 20 days (p=0.05). A reduction in lung neutrophils at 20 days was also seen. Our data shows similar expression patterns of TLR9 in mouse and human lungs, and that barn dust DNA may contribute to inflammation induced following exposure to chicken barn air.

Grant Funding Source: NSERC
Interaction of Epulopiscium Methyl‐Accepting Chemotaxis Proteins with E. Coli CheW and CheA
Authors: Pryde Carson , Piefer Andrew
Published: June 2021

Epulopiscium sp. Type B is a large (up to 250 μm length) unculturable gram positive bacterium that grows symbiotically in the gut of the surgeonfish Naso tonganus. To further understand the unique biology of this organism, we recombinantly expressed GST fusions of Epulo methyl accepting chemotaxis sensory transducers (MCP) in E.coli. MCPs are responsible for sensing and binding extracellular ligands as well as recognizing pH and temperature changes. In E. Coli the MCP interacts indirectly with CheA (sensory kinase) through CheW (coupling protein). The degree of sensory kinase activity controls the motor complex resulting in taxis towards or away from the stimulus. Aligned with the Tar MCP in E. Coli, Epulo MCP 3 displays a 16.2% identity and 28.8% similarity. Epulopiscium MCP 6 has an 18.6% identity and 33.7% similarity. Epulo CheW was aligned with E. Coli CheW and shows 44.9% conservation and 26.9% identity. Epulo CheA has 30.8% identity and 48% similarity when compared with CheA from E. Coli. As a result Epulo MCPs should theoretically interact with CheA through CheW in an in vitro pull‐down assay. These protein interaction studies lay the foundation for future study of Epulo chemotaxis in E. Coli using motility “swarm” assays. This may help to elucidate the environment in which Epulo lives, and its role in symbiosis with the surgeonfish. This project was funded by the Hartwick College Department of Chemistry.
Mechanistic insights into specificity, activity and regulation of Rho exchange factors
Authors: Jaiswal Mamta , Ahmadian Mohammad Reza
Published: June 2021

The multimodular guanine nucleotide exchange factors (GEFs) of the Dbl family mostly share a tandem Dbl homology (DH) and pleckstrin homology (PH) domain organization. The role of this and other domains in the DH‐mediated regulation of the GDP/GTP exchange reaction of the Rho proteins is not well understood. This study presents detailed kinetic data on specificity, activity and regulation of the catalytic DH domains of four GEFs, namely p115, p190, PRG and LARG. We demonstrate that (i) these GEFs are specific exchange factors for the Rho isoforms (RhoA, B, C) and inactive towards other member of the Rho family including Rac1, Cdc42 and TC10; (ii) the DH domain of LARG exhibits the highest catalytic activity, reported for a Dbl protein till now, with a maximal acceleration of 107‐fold (iii) an N‐terminal region of the DH domain is involved in its association with GDP‐bound RhoA monitored by a fluorescently labeled RhoA; (iv) the tandem PH domains of p115 and PRG efficiently contribute to the DH‐mediated nucleotide exchange reaction; (v) in contrast to the isolated DH or DH‐PH domains, a p115 fragment encompassing both the regulator of G‐protein signaling (RGS) and the DH domains, revealed a significantly reduced GEF activity supporting a proposed model of the RGS‐mediated intramolecular autoinhibitory mechanism for p115‐like RhoGEFs. This work was supported in part by the DFG and Forschungskommission der Medizinischen.
Effect of Jobelyn® (Sorghum Bicolor) on nitric oxide and arginase activity in sickle cell anaemic subjects (HbSS)
Authors: ANI ANTHONY AMAECHI , Imaga Ngozi Awa
Published: June 2021

The objective of this in vivo study was designed to investigate the effect of jobelyn on nitric oxide synthase (NOS) and arginase activity in sickle cell anaemic subjects and to elucidate the biochemical pathways through which jobelyn aids in management of sickle cell anaemia; with arginase (EC 3.5.3.1) and nitric oxide synthase (EC 1.14.13.39) pathways under investigation. Both pathways are implicated in vaso‐occlusive crisis usually presented during sickle cell crisis. Three adult human subjects (n=3; Age: 24±3) that had not taken jobelyn prior to the study were recruited from the sickle cell clinic, lagos university teaching hospital. Blood samples were collected before administration, during administration and withdrawal (Day 0, Day7 & 14, and Day 28 respectively) for haematological, biochemistry, nitric oxide and arginase activity analysis. The mean haemoglobin concentration (Hb) g/L on Day 7 and Day 14 were statistically higher than Day 0(P<0.05), while Day 28 is lower than Day 7 and Day 14 (P<0.05). There was no significant difference in nitric oxide concentration μM and arginase activity U/L in all sampling days (P>0.05). In conclusion, administration of jobelyn increases the Hb concentration in sickle cell anaemic subjects, but does not affect nitric oxide concentration and arginase activity.

No support was received.
Site‐specific proteomic approach for study protein S‐nitrosylation
Authors: Ding Shi‐Jian , Liu Miao
Published: June 2021

Protein S‐nitrosylation, the covalent attachment of nitric oxide (NO) to cysteine residues, is an emerging post‐translational modification (PTM) which regulates a large variety of cellular functions and signalling events. Characterization of S‐nitrosylated proteins and their modification sites is essential to understand the action of NO in biological systems. Here we present a novel and robust method for the identification of protein S‐nitrosylation sites in complex protein mixtures. The approach utilizes the Cysteinyl affinity resin to selectively enrich S‐nitrosylated peptides. The enriched samples were analyzed by nanoscale liquid chromatography tandem mass spectrometry on LTQ‐Orbitrap mass spectrometer. We applied this approach to MDA‐MB‐231 cells treated with Angeli's salt. Angeli's salt has been shown to inhibit breast cancer tumor growth and angiogenesis. A total of 177 S‐nitrosylation sites were identified and an S‐nitrosylation motif was revealed in our study. The 177 sites are significantly more than the number reported by previous methods, demonstrating the efficiency of our approach.
Thrombospondin‐1 and Angiotensin‐II inhibit soluble guanylyl cylase through an increase in intracellular calcium concentration
Authors: Ramanathan Saumya , Mazzalupo Stacy , Boitano Scott , Montfort William R
Published: June 2021

Nitric Oxide (NO) regulates vasodilation, wound healing, platelet activation and many other biological processes. The primary target for NO is soluble guanylyl cyclase, a 150 kDa hemeprotein that catalyzes formation of cGMP from GTP. NO binding stimulates the cyclase activity by ~200‐fold. Thrombospondin‐1 (TSP‐1), a secreted matricellular protein, was recently discovered to inhibit NO signaling and sGC activity. TSP‐1 inhibition of sGC requires binding to cell‐surface receptor CD47 using a C‐terminal domain called E3CaG1. We have examined the mechanism behind this inhibitory activity using Jurkat T cells and, using digital imaging microscopy on live cells, show that E3CaG1 binding results in an increase in [Ca2+]i, up to 300 nM. Blocking this increase with the Ca2+ chelator BAPTA overcomes sGC inhibition. Addition of angiotensin II, a vasoconstrictor known to function in part through increasing [Ca2+]i, also strongly inhibits sGC activity. Allosteric stimulators of sGC, YC‐1 and BAY 41‐2272, overcome E3CaG1‐mediated inhibition of NO‐stimulated sGC activity. Taken together, these data indicate that TSP‐1 regulates sGC through a mechanism involving calcium signaling, providing another link between calcium and NO signaling. The mechanism behind this new level of sGC regulation is under investigation.
Successes and challenges in rural Zambia: An interdisciplinary effort in a pro‐vitamin A biofortified maize efficacy trial in preschool children
Authors: Arscott Sara A , Nuss Emily T , Bresnahan Kara A , Tanumihardjo Sherry A
Published: June 2021

Food‐based approaches to increase access to and intake of vitamin A‐rich foods in developing countries show promise as sustainable strategies for combating vitamin A deficiency. Staple foods biofortified with vitamin A must not only prove efficacious, but also effective, by reaching and gaining acceptance by end‐users. A critical step in this process is the successful implementation and community acceptance of efficacy trial interventions. A retrospective examination of a 3‐month biofortified maize feeding trial in rural Zambian preschool children conducted by an international team of researchers was performed. Transcripts of community meetings and observations by the researchers suggest several important factors contributed to community acceptance and low drop‐out, including community input and on‐going sensitization, utilization of local resources, employment of subject's mothers and other community members, investment in local infrastructure and inclusion of a preschool program for the children. Additionally, the success of the trial required an interdisciplinary, integrated effort that included many players, from plant breeders and nutritionists, to teachers, healthcare workers and community leaders. Challenges that were encountered included communication, resource acquisition, anxiety about witchcraft or Satanism and timely staff reimbursement. (Support from HarvestPlus 8217)
Targeted disruption of Pde3b, but not Pde3a, protects murine heart from ischemia/reperfusion injury: Proteomic approaches to possible mechanisms
Authors: Chung Youn Wook , Chen Yong , Gucek Marjan , Murphy Elizabeth , Manganiello Vincent
Published: June 2021

The Pde3 gene family contains two subfamilies, PDE3A and PDE3B, which are generated from distinct, but closely related, genes. Cilostazol, a PDE3 inhibitor, has been reported to protect rabbit and murine heart against ischemia/reperfusion (I/R) injury. Targeted disruption of Pde3b, but not Pde3a, protected the heart from I/R injury, with significantly improved postischemic recovery of left ventricular developed pressure and reduced infarct size.

Cardioprotective signals are thought to be transmitted to mitochondria by specialized vesicular structures, signalosomes, which are assembled in caveolae and lipid rafts and contain various signaling molecules. We therefore performed a comprehensive characterization of ischemia‐induced caveolin‐enriched fractions (ICEF) in mitochondria preparations from Langendorff‐perfused hearts from WT and PDE3B KO mice before/after ischemia, using a combination of subcellular fractionation and isobaric tag for relative and absolute quantitation (iTRAQ). More than 500 ICEF proteins were identified; most of them were mitochondrial. Identified proteins were grouped according to their differential expression: i) no changes before/after ischemia and no differences in wild type and KO hearts, ii) changes after ischemia in both wild type and KO hearts, iii) changes after ischemia only in wild type hearts, and iv) changes after ischemia only in KO hearts. Furthermore, fractions containing the signalosomes were enriched in connexin 43‐containing mitochondrial populations, i.e. subsarcolemmal mitochondria (SSM), not interfibrillar mitochondria (IFM).

In conclusion, these data suggest that targeted inactivation of Pde3b modifies composition of signalosomes associated with SSM to exert cardioprotective effects during I/R, and that PDE3B‐selective inhibitors might provide useful therapeutics to limit I/R injury.
Structural and Kinetic Characterization of NAD Synthetase from H. sapiens and M. tuberculosis
Authors: Chuenchor Watchalee , Chang Kaiti , Doukov Tzanko I. , Gerratana Barbara
Published: June 2021

NAD synthetase catalyzes the ATP‐dependent NAD formation from nicotinic acid adenine dinucleotide in NAD biosynthesis. This enzyme is essential for survival of Mycobacterium tuberculosis but not of humans. In this work, we are determining structural and mechanistic differences between NAD synthetase from M. tuberculosis (NAD synthetaseTB) and from Homo sapiens (NAD synthetaseHS) using x‐ray crystallography and kinetic analysis. NAD synthetaseTB is homooctameric with a 40 Å long inter‐subunit ammonia tunnel that allows transport of ammonia from the glutaminase active site to the synthetase active site. Structural complexes of NAD synthetaseTB with ligands have been solved within a 2.6–3.0 Å resolution, revealing key amino acid residues responsible for substrate binding and catalysis. In addition, NAD synthetaseHS expressed in Sf9 insect cells was co‐crystallized with an intermediate analog for structural comparison. This structure solved at a 3.0 Å resolution using molecular replacement and kinetic characterization are here reported. The crystal structures and kinetic parameters of both NAD synthetaseHS and NAD synthetaseTB can help clarify the basis of substrate recognition and specificity, leading to the design of high affinity inhibitors specifically targeting NAD synthetaseTB.
HIV‐1 Env can pseudotype HTLV‐I and provide envelope‐mediated protection against super‐infection
Authors: Klase Zachary Alan , Jeang Kuan‐Teh
Published: June 2021

Superinfection exclusion is the phenomenon whereby a virus infected cell becomes resistant to subsequent re‐infection with the same or a related virus. Human Immunodeficiency Virus (HIV) is one of the many viruses for which superinfection exclusion has been demonstrated. Three HIV proteins (Nef, Vpu and Env) have been implicated in mediating the effect; however, the mechanism of resistance has not been well described. Here we demonstrate that the HIV‐1 envelope protein (Env) when over‐expressed is sufficient to mediate superinfection exclusion. The mechanism appears to work at the cell surface CD4 receptor and is independent of CD4 down‐regulation and does not appear to involve either the CCR5 or the CXCR4 co‐receptor. These results challenge the assumption that down regulation of the CD4 receptor during infection is critical in preventing super infection. As an extension to our study, we have demonstrated that the Human T‐Cell Leukemia Virus (HTLV) can be pseudotyped using HIV Env. We have developed a replication competent HTLV‐1 virus that expresses the HIV‐1 (in place of the HTLV‐1) Env protein. With this chimeric virus, we plan to transduce CD4+ human T cells and demonstrate that these cells can be protected from HIV‐1 superinfection.

This research was supported by NIAID intramural funds and a GCE phase 1 award from The Bill and Melinda Gates Foundation.
Inhibition of ecto‐ATPase activity by CrATP interferes in the promastigote‐macrophage interaction in Leishmania amazonensis infection
Authors: Moreira Otacilio da Cruz , Vidal Vitor Ennes , d'Ávila‐Levy Claudia Masini , Britto Constança , Barrabin Hector
Published: June 2021

Recent works have shown the relationship between Ecto‐Nucleoside‐Triphosphate‐Diphosphohydrolases (Ecto‐NTPDases) and virulence and infectivity in trypanosomatids. We characterized the inhibition of the ecto‐ATPase activity and promastigote growth of Leishmania amazonensis by CrATP. Furthermore, we used this compound to investigate the role of ecto‐NTPDase in the adhesion and internalization of L. amazonensis in resident peritoneal macrophages from BALB/C mice. CrATP partially inhibits the ecto‐ATPase activity of ecto‐NTPDase, presenting Ki= 575.7±199.1 and 383.5±79.0 μM, in the presence or absence of 5 mM MgCl2, respectively. The apparent Km for ATP (2.9±0.5 mM) is not altered by CrATP, suggesting a reversible non‐competitive inhibition. When CrATP was added to the cultivation medium at 500 μM, it drastically inhibited the cellular growth after 72 h. The interaction of promastigote forms of L. amazonensis with macrophages is strongly affected by CrATP. When the parasites are pre‐treated with 500 μM CrATP before interacting with macrophages, the adhesion and endocytic index are strongly reduced to 53.0±14.8% and 39.8±1.1%, respectively. These results indicate that ecto‐NTPDase plays an important role in the infection process caused by L. amazonensis and that CrATP is a potential prototype for Leishmaniasis chemotherapy.
Structure‐activity optimization of 4H‐pyridopyrimidines: a class of selective bacterial protein synthesis inhibitors
Authors: Bullard James M , Guiles Joseph W. , Toro Andras , Ochsner Urs
Published: June 2021

Introduction

Bacterial protein synthesis is the target for numerous natural and synthetic antibacterial agents. Previous screening of a focused chemical compound library using a polyU mRNA‐directed aminoacylation/translation protein synthesis system from Escherichia coli yielded a lead series of compounds that inhibit protein synthesis with IC50's ranging from 3 to14 μM (Ribble, et. al. (2010) AAC, 54, 4648–4657). This series of compounds all contained the same central scaffold composed of tetrahydropyrido[4,3‐d]pyrimidin‐4‐ol (e.g. 4H‐pyridopyrimidine). These compounds inhibited bacterial growth with minimum inhibitory concentrations (MICs) ranging from 0.25 to 32 μg/ml.

Results

Over 100 analogs of the initial compounds were designed and synthesized and assayed. In biochemical protein synthesis reactions nineteen of the compounds exhibited IC50 values less than 2.0 μM and seven less than 1.0 μM. These compounds were active against S. pneumoniae (MIC=2 μg/mL), H. influenzae (MIC=1 μg/mL), and M. catarrhalis (MIC≤0.12 μg/mL) and S. aureus (4= μg/ml). Macromolecular synthesis (MMS) assays in E. coli and S. aureus confirmed that antibacterial activity resulted from specific inhibition of protein synthesis.

Conclusion

Optimization of 4H‐pyridopyrimidines yields potent inhibitors of protein synthesis with good activity against respiratory pathogens.
Enzymatic and Structural Characterization of a Phosphoglycolate Phosphatase Virulence Factor from Staphylococcus aureus
Authors: Ramirez Sebastian , Hill Jacqueline , O'Handley Suzanne
Published: June 2021

Staphylococcus aureus is a major cause of hospital acquired infection. The multi‐drug resistant nature of certain S. aureus strains makes the discovery of new S. aureus drug targets vital. A newly discovered virulence factor from S. aureus was described as a homolog of NAGD. We cloned the gene, expressed and purified the protein, and determined its activity. This virulence factor is not an ortholog of NAGD UMPase, but rather a phosphoglycolate phosphatase. A phosphoglycolate phosphatase from E. coli has been implicated with a role in DNA repair. If the S. aureus phosphoglycolate phosphatase is involved in both DNA repair and virulence, this would be a novel type of virulence factor, and a potential novel antibiotic target. We have subcloned this PGPase into a His‐tag vector and purified the protein using nickel affinity and size exclusion chromatography. We are currently screening crystals for use in determining the xray crystal structure.
ApaH Diadenosine Polyphosphatase from Legionella pneumophila
Authors: Patron Alejandra Rizo , Thomson Joshua , Baltus Andrew , Edelstein Martha , Zhuang David , Edelstein Paul , O'Handley Suzanne
Published: June 2021

Legionella pneumophila (LP), the causative agent of Legionnaires' disease, is a Gram‐negative bacterium and facultative intracellular parasite. The Nudix diadenosine polyphosphatase, NudA, is a virulence factor in LP, and a related nonNudix diadenosine polyphosphatase, ApaH, is a virulence factor in E. coli and acts additively with the E. coli NudA for virulence. Thus LP apaH was cloned using PCR and an apaH knockout mutation was created in LP. While the LP apaH knockout mutation had no apparent phenotype in studies of growth at 25° C or intracellular virulence, a double apaH/nudA knockout mutant could not be created, likely because this dual mutation is lethal, thus indicating a role for apaH in LP. With this in mind, we subcloned apaH for overexpression to study the enzyme, but the resultant protein was insoluble, so we once again subcloned the gene, this time using the maltose binding protein (MBP) as a solubility tag. We were successful in obtaining a soluble fusion protein with diadenosine polyphosphatase activity. Due to technical difficulties using the MBP as an affinity tag, ApaH is currently being purified using more traditional purification methods including size exclusion and ion exchange chromatography. Once the ApaH diadenosine polyphosphatase has been purified, it will be characterized further enzymatically.
Characterization of Plasmodium falciparum Holo‐ACP Synthase: a Potential Antimalarial Drug Target
Authors: Tami Germaine , Murphy Peter W , Byers David M
Published: June 2021

The protozoan parasite Plasmodium falciparum is responsible for the most lethal form of malaria and kills about one million people annually, most of whom are children in sub‐Saharan Africa. Fatty acid synthesis (FAS) is a validated antimalarial strategy: it occurs in a specialized parasite organelle (the apicoplast) and uses a dissociated type II FAS system similar to bacteria. Bioinformatics analysis indicates that P. falciparum encodes a single enzyme, holo‐ACP synthase (PfAcpS), responsible for activating acyl carrier protein (ACP) involved in FAS. PfAcpS consists of a signal/leader peptide, a large N‐terminal domain of unknown function, and a C‐terminal domain (PfAcpS‐C) that is highly homologous (30–50% similarity) to trimeric bacterial AcpS enzymes. We have expressed and isolated the soluble PfAcpS‐C domain and shown that it catalyzes the transfer of (acyl)phosphopantetheine to PfACP from coenzyme A or from its derivatives acetyl‐CoA and octanoyl CoA. PfAcpS exhibited much lower activity than E. coli AcpS and preferred higher concentrations of CoA derivatives. Several novel inhibitors of E. coli AcpS also blocked PfAcpS activity, although the degree of inhibition differed between the enzymes. The sequence similarity between PfAcpS‐C and bacterial AcpS enzymes suggests that drugs targeting the latter might also inhibit P. falciparum growth and pathogenesis. (Supported by NSERC)
Polymorphic variations affecting response to anti‐VEGF therapy in patients with exudative age‐related macular degeneration
Authors: Wang Vinson Matthew , Rosen Richard , Meyerle Catherine , Kurup Shree , Chew Emily , Chan Chi‐Chao , Tuo Jingsheng
Published: June 2021

Age‐related macular degeneration (AMD) is the leading cause of central, irreversible visual impairment in the elderly and genetic components are major contributors to disease pathogenesis. The most widely utilized therapy option for neovascular AMD is intravitreal administration of anti‐vascular endothelial growth factor (VEGF) antibodies such as bevacizumab and ranibizumab. However, some AMD patients do not respond (continue to exhibit subretinal fluid or continue to loose visual acuity) adequately to this therapy. We hypothesized that a pharmacogenomic mechanism may play a role in the differentiated therapy response and examined 12 SNPs within CFH, HTRA1, IL‐17, IL‐23R, CYP3A, and LEP in 43 patients (29 responders and 14 non‐responders). We observed a higher rate of G allele carriers in the responders (4/29) compared to the non‐responders (0/14) for an IL‐23R SNP, rs11465804. In addition to examining genotypic variations, we also arrayed the expression of 36 cytokines in the plasma of selected responders/non‐responders using R&D Systems' Human Cytokine Array Panel A. We found that plasma IL‐17, a pro‐inflammatory cytokine regulated by IL‐23R, was higher in the non‐responders (n=1) than that of the responders (n=2). These results suggest that gene variation within IL‐23R and altered relevant cytokine profile might contribute to the distinct efficacy of anti‐VEGF therapy for exudative AMD.
The Association Between Food Insecurity And Gait Speed Varied By Smoking Status In Older Adults
Authors: Xu Beibei , Zizza Claire
Published: June 2021

Very few studies have examined the association between food insecurity and physical performance among older adults. Using data from the 1999–2002 National Health and Nutrition Examination Survey, we examined the association between food security and a performance‐based measure, gait speed, in 3,144 older adults (≥ 50 y). Participants were categorized by the US Food Security Survey Module and gait speed (m/s) was calculated as the walking distance (20 ft = 6.15 m) divided by time (s). Multiple linear regression models controlling for age, gender, race/ethnicity, education and income were used. The association between food insecurity and gait speed varied by smoking status (P = 0.005). For non‐smokers, those who were marginally food secure (β = −0.13, P = 0.016) and food insecure (β = −0.10, P = 0.004) had significantly slower gait speeds than food secure participants. No statistically significant associations were observed among current and former smokers. Finding no association among current and former smokers may be a result of poor physical performance among the referent group, food secure participants who smoke.

Grant Funding Source: Alabama Agricultural Experiment Station (ALA013‐020)
Cyclosporin A and itraconazole synergistically inhibit endothelial cell proliferation and tube formation
Authors: Nacev Benjamin Alexander , Liu Jun O
Published: June 2021

Angiogenesis underlies a number of pathologies including cancer and macular degeneration, but current single agent antiangiogenic therapies are only modestly effective in the treatment of these diseases. Thus, we sought to identify compounds which combinatorially and synergistically inhibit angiogenesis. From a screen of 741 binary clinical drug combinations for the synergistic inhibition of endothelial cell proliferation we identified cyclosporin A (CsA), an immusuppressant, and itraconazole, an antifungal drug. The IC50 dose of each drug is reduced by 3 to 9 fold in combination. The combination also synergistically inhibited endothelial cell tube formation and was specifically active against endothelial cells when compared with human foreskin fibroblast viability as well as Jurkat T cell and HeLa cell proliferation. We conclude that it is a viable strategy to combine existing clinical drugs to synergistically inhibit endothelial cell proliferation. Drawing on a pool of clinical drugs has the potential advantage of a faster transition to phase II studies when potent combinations are identified. This model may be useful for the development of new antiangiogenic treatments. This work was supported in part by NCI, FAMRI, CTSA, the Keck Foundation, the Walsh Prostate Cancer Fund, and the Commonwealth Foundation (J.O.L.), and by the NIH Medical Scientist Training Program (B.A.N).
Effectiveness of dietitian‐led diabetes management program on glycemic and diet control after a 2 year intervention in primary care in Taiwan
Authors: Sung Yi‐ching , Lin Chia Hui , Pai Hsiu‐hua , Hsu Chih‐Cheng , Shin Shyi Jang , Huang Meng‐Chuan
Published: June 2021

Objective

The aim of this study was to evaluate effectiveness of dietitian‐led diabetes management program on glycemic and diet control after a 2 year intervention in primary care clinics in Taiwan.

Methods

At baseline, we randomly assigned adult patients with type 2 diabetes to a routine care control (n=75) or a dietitian‐led intervention group (n=79) receiving diabetic self‐management education every 3 months.

Results

After 2 year intervention, completion rates were 77% (control: n=43; intervention: n=48). Hemoglobin (HbA1c %) were significantly different between baseline and 2 year only in intervention patients with baseline HbA1c ¡Ù7%, changes of HbA1c also correlated significantly (r=0.365, p=0.016) with carbohydrate intake. However, HbA1c did not improve significantly in control subjects regardless of entry HbA1c levels.

Conclusion

On‐site dietitian‐led management of diabetes can improve glycemic control in patients with poorly managed type 2 diabetes in primary care setting in Taiwan.
Case Control Study for the Evaluation of Beneficial Effect(s) of Pistachio Nut Intake on Cardiovascular Risk Factors in Asian Indians with the Metabolic Syndrome
Authors: Gulati Seema , Misra Anoop
Published: June 2021

Background

The metabolic syndrome is precursor of diabetes and coronary heart disease. Previous studies have suggested that nut consumption leads to improvement in risk factors of the metabolic syndrome. Pistachio nuts are easily available and frequently consumed by general population.

Objective

To determine whether substituting 15% and 20% of the daily caloric intake in the form of pistachio nuts improves the risk factors of the metabolic syndrome among Asian Indians.

Design

Randomized case control design.

Setting

Outpatient dietary modification, counseling and blood analysis.

Patients

Sixty patients with the metabolic syndrome.

Methods

The intervention duration was of 24 weeks preceded by a diet and exercise run‐in period of 30 days. For initial 12 weeks subjects in case group were given basic diet plus pistachio nuts constituting 15% of daily calorie intake. Subsequently, dosage of pistachio was increased to 20% of the total calorie intake for another 12 weeks. Controls were given equi‐calorie, high fiber low fat diet. All the subjects in cases and controls were followed for 6 months. At the end of the study, comparative analysis was done between cases (who consumed pistachio nuts) and controls for anthropometry and measures of obesity, measures of glycemia, anti‐inflammatory markers and markers of oxidative stress.

Results

Results would be presented after analyzing the data.

Conclusions

Results are awaited.
Environmental change program associated with relative decrease in frequency of carryout/fast food consumption among African American caregivers
Authors: Cessnun Kathryn , Dennisuk Lauren , Coutinho Anastasia , Sharma Sangita , Shin Ahyoung , Christiansen Karina , Yong Rachel , Lwin Julia , Gittelsohn Joel
Published: June 2021

Low‐income African American families in Baltimore City live in neighborhoods with limited access to healthy foods. The Baltimore Healthy Eating Zones (BHEZ) study aimed to improve diet quality among adolescents and their caregivers through an environmental intervention that involved peer mentoring, cooking demonstrations, taste tests, point‐of‐sale marketing and promotional giveaways at recreation centers and corner stores over a 9 month intervention period. Caregiver and child dyads (n=242) from 7 control and 7 intervention sites were interviewed before and after the intervention and asked about their monthly food purchases from carry out and fast food restaurants. Changes in the frequency of these food purchases were analyzed using a Wilcoxan Signed Ranks Test. The frequency of food purchases by caregivers from carry out and fast food restaurants significantly increased from 1.50±2.18 to 3.88±4.99 times per month among the control group (p=0.037) but did not significantly change from 8.67±10.06 to 6.12±8.70 times per month in the intervention group (p=0.119). These preliminary findings suggest that an environmental intervention like the BHEZ program may be an effective approach for improving the food purchasing behaviors of adult African American caregivers.

Grant Funding Source: The Robert Wood Johnson Foundation
Lifestyle barriers to managing hypertension in people of African descent
Authors: Lancaster Kristie J , Midberry Sara A , Watts Sheldon O , Cole Helen V , Schoenthaler Antoinette M , Ogedegbe Gbenga
Published: June 2021

Purpose

To determine the lifestyle behaviors that act as barriers to controlling blood pressure among a sample of hypertensive persons of African descent.

Methods

A total of 25 people of African descent diagnosed with hypertension (HTN) were recruited at 3 churches, one African American (AA, N=10), one African immigrant (AI, N=7), and one predominantly West Indian (WI, N=8). A Nominal Group Technique (NGT) focus group session was held at each church. Each group generated a list of responses to the question: “What are some things that make it hard for people to follow lifestyle recommendations to take care of their high blood pressure?” Participants then ranked their top responses, and the rankings were summed. The 10 responses with the highest scores were then given a rating of 0 to 10 by participants and the ratings were summed.

Results

The WI group was older, had less education, and had had HTN for longer. The AI group was the youngest, had the most education, and had HTN for the shortest time. The AA & WI groups rated stress, physical limitations to exercise, and weight maintenance in their top 5 barriers. The AI group rated time for exercise, work, and eating habits in their top 5.

Conclusions

This study suggests groups of African descent may have different lifestyle barriers to managing HTN. Future work should examine the need for tailoring intervention by ethnic group. Funded by NHLBI.
Fish oil increases tumor associated T regulatory cells in experimentally induced colitis
Authors: Gurzell Eric A , Rondini Elizabeth A , Fenton Jenifer I
Published: June 2021

Our laboratory recently reported the unexpected observation that feeding fish oil enriched with docosahexaenoic acid (DFO) exacerbated colitis and colonic dysplasia in SMAD3−/− mice. These effects were associated with reduced T cell populations in secondary lymphoid tissue. The objective of this study was to determine the effect of DFO on immune cell populations and markers of inflammation at the local site of inflammation/dysplasia by immunohistochemistry. Mice were fed AIN‐93G diets modified to include corn oil (CO), safflower oil (SF), or DFO (0.75–6.00%) as the fatty acid source for 8 wks, gavaged with H. hepaticus to induce colitis, then sacrificed 4 wks post‐infection. We found significant differences between dietary treatments for multiple T cell markers in the proximal colon. Compared to controls, total T lymphocytes as well as FoxP3 positive cells were more prominent in the 3.75 and 6% DFO diets, which corresponded with the highest colitis scores. Staining for iNOS and COX‐2, markers associated with colon carcinogenesis, did not differ across treatments. These data suggest that DFO promotes an immunosuppressive environment at the tumor site with strong direction by T regulatory cells, shifting from a Th1 type to Th2 type immune response. The observed changes in immune cell populations and suppressed cell‐mediated immunity may begin to explain the exacerbated colitis.

Grant Funding Source: n/a
Tea and coffee and basal cell carcinoma in a case‐control study
Authors: Ferrucci Leah M , Cartmel Brenda , Molinaro Annette M , Leffell David J , Bale Allen E , Mayne Susan T
Published: June 2021

Tea and coffee may impact carcinogenesis via their bioactive components, such as polyphenols and caffeine. Basic science research suggests a protective role for several of these compounds in relation to skin cancer, but epidemiologic data are limited. We evaluated the association between tea and coffee consumption and basal cell carcinoma (BCC) risk in 685 non‐Hispanic Whites under age 40 in a case‐control study in Connecticut. BCC cases (n=329) were identified through Yale's Dermatopathology (YDP) database. Controls (n=356) were randomly sampled from individuals in YDP's database with benign diagnoses and frequency matched to cases on age, gender, biopsy year, and biopsy site. Subjects completed a structured in‐person interview and provided a saliva sample. We calculated multivariate odds ratios (OR) and 95% confidence intervals (CI) with unconditional logistic regression models. There was no clear association between regular consumption of tea (OR=0.94, 95% CI=0.66–1.35) or caffeinated coffee (OR=0.82, 95% CI=0.55–1.22) and BCC. Amount, frequency, and duration were not associated with risk. There were suggestive differences by tea type, with opposing associations for green (OR=0.86, 95% CI=0.54–1.36) and black (OR=1.26, 95% CI=0.87–1.83) tea, but neither reached statistical significance. There was no evidence of effect modification by gender, single vs. multiple BCC, or melanocortin 1 receptor variant status. Overall, our findings do not support a clear association for tea or coffee and BCC. Funded by National Cancer Institute grants 1P50 CA121974 and 1F32 CA144335.
Dietary carbohydrate intake, plasma adipokine concentration and risk of breast cancer
Authors: Shin Hye Lim , Kim Jeongseon , Sung Mi‐Kyung
Published: June 2021

Dietary carbohydrate intake and obesity are suggested as risk factors for postmenopausal breast cancer. The study was conducted to determine whether carbohydrate quality measured by glycemic index(GI) and glycemic load(GL), and plasma adipokine concentration account for breast cancer risk. Breast cancer patients (n=263) and hospital‐based control subjects (n=263) were recruited from National Cancer Center, Korea. GI and GL were calculated based on food frequency questionnaires, and plasma concentrations of adiponectin and leptin were determined. Study results indicated that the GI(p=0.037) and GL(p=0.045) values were negatively associated with plasma adiponectin concentration while GL (p=0.028) was positively associated with plasma leptin concentration. There was a significant association between GI (OR = 6.06, 95% CI=2.24–16.37, p=0.0029), and GL (OR=4.42, 95% CI=1.59–12.28, p=0.0191) with breast cancer risk in postmenopausal women. The associations between GI, GL and breast cancer were more evident in subjects with lower plasma adiponection. This study was funded by Korean Science and Engineering Foundation (R01‐2007‐000‐11293‐0).
Dietary Intake and Serum Levels Status of Folate and Vitamin B12 among Jordanian Colorectal Cancer Patients: a case‐control study
Authors: Waly Mostafa I , Arafa Mostafa A , Jriesat Sahar B , AlKhafajei Ahmed M , Sallam Sunny A
Published: June 2021

Low daily dietary intakes of folate and vitamin B12 have been associated with an increased risk of colorectal cancer in animal and human studies. The retrospective dietary intake of folate and vitamin B12 was investigated as well as the biochemical measurements of these nutrients in colorectal cancer patients and controls group in Jordan. Four hundred and forty subjects were recruited for this case‐control study, 220 colorectal cancer patients and 220 controls. The mean daily intake of folate and vitamin B12 was significantly lower amongst patients (189.2 ± 11.7 μg/day and 1.09 ± 1.1 μg/day respectively) when compared to the controls group (212.5 ± 13.1 μg/day and 1.49 ± 1.3 μg/day respectively). The same pattern was observed for folate serum measurements of colorectal cancer patients (6.6 ± 0.3 μg/L) as compared to controls (16.2 ± 2.8 μg/L) and for serum vitamin B12 of cases (233.7±17.9 pg/mL) as compared to controls group (458.9±30.9 pg/mL). This study suggests that the studied colorectal cancer patients had a low dietary intake of folate and vitamin B12 that is concomitant with low serum levels of these two nutrients, a well established etiological factor for different types of human cancer, including colorectal cancer.
Associations of the thymidylate synthase 3¡¦UTR polymorphisms with one carbon status, lymphocytic p53 oxidative lesions and hepatocellular carcinoma progression
Authors: Kuo Chang Sheng , Hwung Chyi‐Yun , Kuo Shine‐Tou , Lin Ching‐Yih , Cheng Chin‐Pao , Chen Chien‐Hung , Yang Feili Lao , Huang Rwei‐Fen Syu
Published: June 2021

The aims of the study were to investigate relationships between thymidylate synthase (TS) 3'UTR polymorphisms. Genotypes in a 6‐bp deletion/insertion in the 3’‐untranslated region (TS 3'UTR), blood folate and homocysteine (Hcy) levels were determined on 232 HCC patients. The frequencies of TS 3’‐UTR genotype of the study HCC patients were 52% (del6/del6), 43% in del6/ins6, and 5% in ins6/ins6. Increased frequencies of one 6‐bp insertion in the TS genotypes were associated with decreased Hcy levels (r = −0.154, P = 0.04). Compared with the (del6/del6) variant, HCC patients with at least one 6‐bp insertion variants had lower levels of AFP tumour marker (P = 0.05). Among individuals with low serum folate levels (< 14 nM), the TS del6/ins6 and ins6/ins6 variants had significantly lower levels of p53 promoter oxidative damage than the del6/del6 variant. After adjustment for multiple factors including life‐style, clinical factors and serum folate status, the odds ratios for developing large tumour (size > 3 cm) were 0.24 (95% CI: 0.07–0.08) for combined MTHR C677T mutant variants (CT+TT genotypes) and TS 3'UTR mutant variants (del6/ins6 + ins6/ins6) compared with wild type variants. This protective effect against developing large tumour was negated after further adjustment of lymphocytic p53 oxidative lesions. Taken together, our data suggested that HCC patients carrying combined genotypes with at least one mutant allele in MTHFR C677T and TS 3'UTR 6 bp‐insertion variants have reduced risks of developing large tumour, which may be mediated by amelioration of p53 oxidative lesions.

Grant Funding Source: Department of Health in Taiwan
The effects of omega‐3 fatty acids and bexarotene on human breast cancer progression
Authors: Hawk Susan Nicole , Trappmann Jessica
Published: June 2021

Breast cancer cell growth can be inhibited by retinoid X receptor (RXR) specific retinoids. Omega‐3 fatty acids share growth regulatory effects similar to those noted when breast cancer cells are treated with RXR specific retinoids. One synthetic RXR‐selective retinoid ligand, bexarotene (LCD 1069, Targretin), is an effective antineoplastic agent in rodent mammary carcinoma models. It is also used clinically to treat cancer patients. Of concern is that some patients are unable to tolerate high doses of treatment drugs. We hypothesized that omega‐3 fatty acids and bexarotene may work synergistically to slow breast cancer cell growth. To facilitate this study, we used an in vitro cell culture model to investigate the relationship between α‐Linolenic acid (ALA), Docosahexaenoic acid (DHA) and Eicosapentaenoic acid (EPA) alone and in conjunction with bexarotene on MCF‐7 cell growth. Following a 72 hr incubation, neither bexarotene nor ALA altered cell growth. DHA alone and in combination with bexarotene showed a strong growth inhibitory effect. EPA alone was not as effective in altering cell growth. Interestingly, when combined with bexarotene, EPA tended to slow growth more that when cells received EPA alone. Thus, select omega‐3 fatty acids are most effective in slowing breast cancer cell progression, and may even enhance the uptake of retinoid treatment drugs and help potentiate their efficacy.
Rosmanol potently induces apoptosis through both the mitochondrial apoptotic pathway and death receptor pathway in human colon adenocarcinoma COLO 205 cells
Authors: Cheng An‐Chin , Lee Ming‐Fen , Pan Min‐Hsiung , Hong Wei‐Hsiang , Kuo Chia‐Chang
Published: June 2021

The roles of tumor suppression of rosemary (Rosmarinus officinalis), a culinary spice and medicinal herb, have been attributed to the major components, including carnosic acid, carnosol, and rosmarinic acid, rosmanol, and ursolic acid. This study was to explore the effect of rosmanol on the growth of COLO 205 human colorectal adenocarcinoma cells and to delineate the underlying mechanisms. When treated with 50 μM of rosmanol for 24 h, COLO 205 cells displayed a strong apoptosis‐inducing response with a 51% apoptotic ratio (IC50 ~42μM). Rosmanol increased the expression of Fas and FasL, led to the cleavage and activation of pro‐caspase‐8 and Bid, and mobilized Bax from cytosol into mitochondria. The mutual activation between tBid and Bad decreased the mitochondrial membrane potential and released cytochrome c and apoptosis‐inducing factor (AIF) to cytosol. In turn, cytochrome c induced the processing of procaspase‐9 and procaspase‐3. These results demonstrate that the rosmanol‐induced apoptosis in COLO 205 cells is involvement of caspase activation and involving complicated regulation of both the mitochondrial apoptotic pathway and death receptor pathway.
GP‐1 induces autophagy of human colorectal cancer cells by inhibiting AKT/mTOR and activating type III PI3K/beclin 1 signaling pathways
Authors: Su Chun‐Li , Yen Cheng‐Hsin , Lin Chun‐Nan , Jiang‐Shieh Ya‐Fen , Won Shen‐Jeu
Published: June 2021

GP‐1, a pure compound isolated from the pericarp of Garcinia subelliptica, is a traditional remedy which exhibits anti‐oxidative, anti‐inflammatory, and anti‐tumor effects. Our unpublished data indicate that GP‐1 induced apoptosis of human colorectal cancer HT‐29 cells via caspase‐ and mitochondria‐related pathways. Here we report that GP‐1 also induced autophagy of the cells characterized by formation of acidic vesicular organelles using flow cytometry, expression of autophagic marker LC3‐II using immunoblotting and confocal microscopy, and accumulation of autophagic vacuoles using transmission electron microscopy. In the GP‐1‐treated cells, the expression of p‐AKT, p‐mTOR, and p‐p70S6k was decreased but that of type III PI3K, becline 1, and Atg5‐Arg12 was increased. Pre‐treatment of the cells with autophagy inhibitor 3‐methyladenine or general caspase inhibitor Z‐VAD‐FMK respectively reduced GP‐1‐induced apoptosis or autophagy using annexin V‐FITC and propidium iodine double staining or acridine orange staining followed by flow cytometry. These findings describe the molecular actions of GP‐1 in HT‐29 cells and reveal the enhancement of apoptosis by autophagy, and vice versa.

(Supported in part by National Science Council, Taipei, Taiwan, No. NSC 95‐2313‐B‐309‐001 and NSC 96‐2313‐B‐309‐001‐MY2).
Volatile Components of Mustard Seed induce apoptosis through both caspase‐dependent and ‐independent pathways in human colon cancer cells
Authors: Kim Jaeyong , Hong Hyeran , Lee Juhye , Jeong Ilyun , Shim Kihwan , Seo Kwonil
Published: June 2021

Mustard seed is used widely as a culinary spice because of the taste associated with the isothiocyanates which are responsible for several beneficial health effects. To investigate the anticancer activity of mustard seed in HT‐29 colon cancer cells, the volatile components including 3 types of isothiocynate from mustard seed were collected by simultaneous distillation extraction method. The volatile components induced apoptotic cell death significantly, resulting in a morphological change, apoptotic DNA fragmentation, and an increase in the sub‐G1 apoptotic cell population. The components triggered the mitochondrial caspase‐dependent apoptotic pathway as shown by an increase in Bax/Bcl‐2 ratios, activation of caspase‐9 and caspase‐3, and cleavage of PARP. In addition, the volatile components also induced AIF release from mitochondria, indicating a stimulated apoptosis through caspase‐independent pathway. These results demonstrated that the volatile components from mustard seed inhibited proliferation of HT‐29 cells through inducing apoptosis, which may be mediated via both caspase‐dependent and ‐independent apoptotic pathways.