bluuebunny/crossref_metadata_2025_split huggingface dataset:
Non‐Structural (NS) proteins of Respiratory Syncytial Virus (RSV) target multiple components of the interferon induction and response pathways of innate immunity
Authors: Swedan Samer Fuad , Barik Sailen 
Published: June 2021

RSV is a Pneumovirus of the family Paramyxoviridae. RSV is a leading cause of respiratory disease in the very young and the elderly. Our research focuses on the ability of RSV to inhibit innate immunity. We hypothesize that the two Non‐Structural (NS1 and NS2) proteins of RSV interfere with key signaling components in the interferon (IFN) induction and response cascades, thus inhibiting innate immunity and leading to disease. Our research goals are: 1) To identify key host targets of NS proteins and 2) To identify the IFN‐suppressor domains of the NS proteins. Our preliminary data reveal that NS1 is required for decreasing levels of TRAF3 and IKKε, which are key components for the IFN induction cascade, and that NS2 is required for the degradation of STAT2, a key component for the IFN response cascade. The C‐terminal 20 amino acids (AA) of NS1 were not required for its effect on TRAF3, whereas the C‐terminal 10 AAs were required for the reduction of IKKε. The C‐terminal 4 AAs of NS2 were required for its ability to degrade STAT2. Finally, we discovered that NS1 and NS2 can interact in vitro and can individually interact with the host MAP1B protein.Sponsored by Alabama EPSCoR and NIH grant AI059267
Inhibition of nucleosome assembly by the mismatch repair system
Authors: Takahashi Tatsuro , Taki Kanae , Higashi Torahiko , Nakagawa Takuro , Masukata Hisao 
Published: June 2021

The mismatch repair (MMR) system post‐replicatively corrects replication errors such as base‐base mismatches. The MMR reaction begins with recognition of the errors by the MutS complex. MutS and other factors then identify the erroneous newly synthesized strand. The nature of strand discrimination signals in eukaryotes remains elusive, but a gap or 3′‐end of DNA can work as the signal in vitro. Concurrent with the MMR reaction, nucleosomes re‐assemble on the replicated DNA strands. Nucleosome assembly might impede the MMR reaction, for example by obscuring strand discrimination signals. However, how the MMR system deals with nucleosome assembly remains highly unclear.Using Xenopus egg extracts as a vertebrate model system, we investigated functional interaction between the MMR reaction and nucleosome assembly. In the nucleoplasmic extracts of Xenopus eggs, nucleosomes rapidly assemble on naked DNA. Surprisingly, nucleosome assembly was severely inhibited when a base‐base mismatch was present on the DNA molecule. The MutS complex specifically bound to the mismatch‐containing DNA, and this complex was essential for both repair of the mismatch and inhibition of nucleosome assembly. These results suggest that nucleosome assembly at replication error loci is temporally inhibited by a MutS‐dependent mechanism during the MMR reaction. We propose that this mechanism is important for eukaryotic MMR in the context of chromatin.
Acetylation of Dna2 and Fen1 by P300 Promotes Long Flaps Favoring DNA Stability
Authors: Balakrishnan Lata , Stewart Jason A , Polaczek Piotr , Campbell Judith L , Bambara Robert A 
Published: June 2021

Flap endonuclease 1 (FEN1) and Dna2 endonuclease/helicase (Dna2) sequentially coordinate their nuclease activities for efficient resolution of flap structures that are created during the maturation of Okazaki fragments and repair of DNA damage. FEN1 can be post‐translationally modified by the transcriptional coactivator, p300. Acetylation of FEN1 by p300 inhibits its endonuclease activity impairing flap cleavage, a seemingly undesirable effect. We now show that p300 also acetylates Dna2. However, in contrast to FEN1, acetylation of Dna2 stimulates its 5′–3′ endonuclease, the 5′–3′ helicase and DNA‐dependent ATPase activities, allowing more efficient processing of long flaps. Furthermore, acetylated Dna2 binds its DNA substrates with higher affinity. Dna2 and FEN1 form a complex with p300 both in vitro and in vivo. Differential regulation of the activities of the two endonucleases by p300 indicates a mechanism in which the acetylase promotes formation of longer flaps in the cell at the same time as insuring correct processing. Such intentional formation of longer flaps mediated by p300 in an active chromatin environment would increase the re‐synthesis patch size, providing increased opportunity for incorrect nucleotide removal during DNA replication and damaged nucleotide removal during DNA repair.This work was funded by the National Institute of Health grant GM024441 to R.A.B.
Subunit exchange and DNA‐binding dynamics of Escherichia coli single‐stranded DNA binding protein (SSB)
Authors: Mason Claire Elise , Jergic Slobodan , Dixon Nicholas , Beck Jennifer 
Published: June 2021

Single‐stranded DNA binding proteins (SSBs) are ubiquitous in all organisms, and are responsible for binding to and maintaining the structural integrity of single‐stranded DNA (ssDNA) produced during DNA replication, recombination and repair processes. Escherichia coli SSB is a homotetrameric protein, and its interaction with ssDNA has been well‐characterised in vitro, but the mechanism by which it is able to move rapidly upon DNA during dynamic processes such as replication is largely unclear. In this work, nanoelectrospray mass spectrometry was used to investigate aspects of the E. coli SSB‐ssDNA interaction. By examining subunit exchange between unlabeled SSB and uniformly labeled 15N‐SSB, it was found that ssDNA binding has a strongly inhibitory effect on subunit exchange, which occurs slowly but extensively in the absence of DNA. Furthermore, using mass spectrometry and surface plasmon resonance, intact SSB tetramers, under certain conditions, were found to be able to transfer directly between discrete ssDNA strands. Whether or not transfer occurs is dependent on both oligonucleotide length and salt concentration. This work shows that direct transfer is a possible mechanism of SSB mobility on DNA.This work was funded by an Australian Research Council Discovery Grant
Post‐transcriptional Control of Human Interleukin‐3 Mediated by its 3′‐UTR
Authors: Hernández‐Pérez Marimar , Almodov Natalia , Gonzalez‐Feliciano Jose A , Gonzalez Carlos 
Published: June 2021

Human interleukin‐3 (IL‐3) is a cytokine that stimulates the growth and differentiation of early lymphoid stem cells and has been implicated in certain types of cancer. IL‐3 is a member of a class of transiently expressed mRNAs that harbor Adenosine/Uridine‐Rich Elements (ARE) in their 3′‐UTRs. The 3′‐UTRs of many cytokines play a role in post‐transcriptional control by altering mRNA stability and/or translation. Transfection assays using HeLa and Jurkat cells harboring a luciferase‐IL‐3 3′‐UTR reporter showed a significant reduction in luciferase activity. The mRNA levels of these luciferase chimera do not show any significant differences in HeLa and T‐cells, suggesting that the hIL‐3 3′‐UTR mediates post‐transcriptional control. To understand how the AREs from the hIL‐3 3′‐UTR are involved in controlling translation, we conducted site‐directed mutagenesis of four ARE clusters that were identified in the hIL‐3 3′‐UTR. Firefly luciferase reporters harboring ARE mutations were transfected into HeLa and T cells. The luciferase chimeras lacking the hIL‐3 ARE showed an increase in luciferase activity. Electrophoretic mobility shift assays (EMSA) demonstrated that TIA‐1 and HuR, two ARE‐BP previously associated with translational control, bind to the hIL‐3 ARE. These studies will help delineate the functionality of AREs in the translational control of human IL‐3 and its relationship with oncogenesis.Supported by grants from NIH to C.I.G. (KO1 HL‐04355‐05, U54 CA96297, P20 RR 016174). M.H. is supported by the RISE Program, R25GM061838.
Non‐Coding ncRNA Modified by Environmental Factors and the Epigenetic Code: Translating the Entangled Cancer‐Angiogenesis‐Tolerance Epigenome
Authors: Wissler Josef H. 
Published: June 2021

OBJECTIVEFrom the genome of same genotype, genetic [Mendelian] and epigenetic [non‐Mendelian] information may be translated into different organized or mess‐chaotic [tumor] phenotype variations. ncRNA functions of cells activated by extrinsic [environmental] factors were investigated whether chance or necessity for structural codes govern genomic episcription [Ann. N.Y. Acad. Sci. 1022: 163–184, 2004; 1137: 316–342, 2008].RESULTSNascently formed modified, edited, redox‐ and metalloregulated functional small hairpin ncRNA [<200n] as bioaptamers in RNP complexes were sequenced. Some address conserved homologous helix‐nucleating domains shared in epigenetic regulator proteins entangled in tolerated growth, vascularization, metabolic syndromes, cancer and epigenome indexing. At variance to usual interpretation by genetic complementary and triplet base codes, ncRNA are not “non‐ coding”, if read in another “language alphabeth” [complementary biomolecular imprint interaction]. This epigenetic code consists of different associated intrinsic and extrinsic interactive imprints and factors: [1] Non‐Mendelian nucleic acid 3D‐episcripts in helix‐ nucleating complementary interaction with [2]: Conserved homologous Mendelian domains in epigenetic regulator protein and nucleic acid matrices. [3] Extrinsic and intrinsic factors upon which formation of [1] and interaction with [2] depends [e.g. redox‐and metalloregulation].CONCLUSIONSThe epigenetic code has more diversity, complexity and plasticity repertoires than genetic codes. It implies some chance and heuristic reactions in algorithmic [necessity] processes and Darwin‐Mendel's genetic principles in synergism with some environmental [Lamarck's] influences for epigenetic imprinting and inheritance.
Metabolomic evaluation of Masseter muscle in an experimental malocclusion model
Authors: Farinha Rodrigo , Gomes Marcolino , Tavares Ludgero , Carvalho Rui , Silvério Cabrita A. M. 
Published: June 2021

Occlusal disarrangements are very often in the basis of the sintomatology related with temporomandibular disfunction, which courses with muscular and articular involvement.The aim of this study is to give a contribution to the understanding of metabolic changes in the masseter muscle in an experimental model of malocclusion.20 Wistar male rats, 12 weeks old were distributed in two groups: the control group, kept in the standard conditions and the test group. In the test group we created a change of the normal occlusal contacts by adding fotopolimerized resin on the left superior teeth. After two weeks, all animals were killed and the complete necropsy was performed. Fractions of masseter muscle were collected for histopathological analysis and for NMR analysis. The fragments for NMR analysis were snap‐frozen in liquid nitrogen and stored at −80ºC until the spectroscopy was performed.After the spectra and statistical analysis we observed that Taurine (3.261 PPM); β‐glucose (3.42 PPM); Creatin (3.93 PPM) and a metabolite at 4.334 PPM were significantly changed between the control and test groups on left masseter. Taurine (3.261 PPM), β‐glucose (3.422 PPM), and two metabolites on 4.964 and 1.482 PPM were significantly changed between the control and test groups on right masseter. We didn't observe any statistically significant changes between left and right masseter metabolic profile on control groupThe results of this study suggest that occlusal interferences can produce metabolic changes on masseter muscle on the working and non‐working sides. Taurine and β‐glucose suffered statistically significant changes on both left and right masseter muscles.
Oxidation‐ or Radiation‐Derived Effects on Human Mitochondria as Seen in a Fibroblast Cell Line Derived from a Family Known to Have Elevated Levels of the 4977‐bp Common Deletion in its Mitochondrial DNA
Authors: Stoll Zachary , Pogozelski Wendy 
Published: June 2021

DNA in the mitochondria of cells undergoes deletion events that compromise energy function but do not prevent cells from being viable. The mechanism of how these deletions occur and how they affect cell function are topics of interest. We have been investigating the role and induction of large‐scale deletions in oxidative stress using a variety of methods, including gamma radiation. We observed a dose‐dependent increase in the levels of a 4977‐bp deletion called the “common deletion.” using real‐time PCR with an external DNA quantification standard in fibroblast cells. We are currently manipulating cell conditions to alter the metabolism of cells and compare the effects with and without this deletion.This research was sponsored by the Camille and Henry Dreyfus Foundation, the Geneseo Foundation and the Geneseo Student Association.
Endocytosis of pro‐apoptotic twin‐base RGDSK helical rosette nanotubes in human U937 macrophage cell line
Authors: Singh (Suri) Sarabjeet , Fenniri Hicham , Hemraz Usha , Singh Baljit 
Published: June 2021

Twin‐base helical rosette nanotubes (RNTs), a novel class of biologically inspired nanotubes, hold tremendous potential as targeted drug delivery shuttle. To realize biomedical potential of RNT, there is a critical need to understand the interaction of RNT with the cells. Therefore, we synthesized hybrid RNT that composed of RGDSK and FITC to gain ability to track them during in vitro experiments with human differentiated macrophages. Scanning electron microscopy confirmed nano‐scale dimensions of RGDSK RNTs. FITC‐RGDSK RNT was co‐localized with integrin avb3‐Cy5 on the macrophages within 2 minutes of incubation at 37 deg C. Integrin avb3 and flotallin‐1, a lipid rafts protein, showed fever co‐localization in non‐stimulated cells. FITC‐RGDSK RNT also co‐localizes with EEA1, an early endosome marker. The inhibition of the co‐localization of FITC‐RGDSK RNT with the integrin and translocation into endosomes at 4 deg C demonstrated energy‐dependence of the process. The incubation of differentiated macrophages with RGDSK‐TBL RNT (>5 μM) resulted in nearly 100 % apoptosis within 12 hours. These results suggest that the FITC/RGDSK‐TBL RNT could be used as a versatile platform to deliver a variety of biologically active molecules for cancer therapy.Support: NSERC, AARI.Grant Funding Source: AARI, NSERC
Microbiological evaluation of 3‐((E)‐(4‐chloro‐6‐hydroxypyrimidin‐2‐ylimino)methyl)‐4‐chloro‐2H‐chromen‐2‐one
Authors: Govori Sevdije R , Spahiu Sabrije , Haziri Arben , Daci Majlinda , Sehabi Muhamet , Kurteshi Kemajl 
Published: June 2021

3‐((E)‐(4‐chloro‐6‐hydroxypyrimidin‐2‐ylimino)methyl)‐4‐chloro‐2H‐chromen‐2‐one 3 is synthesized in very good yields by reaction of corresponding 4‐chloro‐2‐oxo‐2H‐chromene‐3‐carbaldehyde 1 and 2‐amino‐6‐chloropyrimidin‐4‐ol 2. Antibacterial activity of products 3 is investigated and results are submitted for their activities against Escherichia coli and Enterochoaco. Applying the disc method we mesasured diameters of the inhibition zone around disc which are previously wetted with N,N‐DMF solution of compounds 3 in concentration 1 mg/l, 0.5 mg/l and 0.1 mg/l.The compounds 3 showed strong activity in reducing the microbial growth. The synthesized compound 3 showed most effective antibacterial activity against Escherichia coli.
γ‐Tocopherol supplementation improves postprandial vascular endothelial function in lean and obese men by decreasing oxidative and nitrative stress
Authors: Mah Eunice , Ballard Kevin D , Kawiecki Diana M , Volek Jeff S , Bruno Richard S 
Published: June 2021

Hyperglycemia induces oxidative stress and inflammation that impairs vascular endothelial function (VEF). We hypothesized that γ‐tocopherol (γ‐T) supplementation would improve hyperglycemia‐mediated decreases in VEF by reducing oxidative stress and inflammatory responses. A randomized, crossover study was conducted in lean and obese men (n=8/group) who followed their usual diet or were also provided γ‐T‐rich capsules (500 mg/d, 5 d) prior to an oral glucose challenge. Brachial artery flow‐mediated dilation (FMD) and markers of oxidative/nitrative stress and inflammation were measured throughout the 3 h postprandial period. Supplementation increased plasma γ‐T by 170% in all men without affecting α‐T. Baseline HOMA was greater in obese men and correlated to FMD, but was unchanged by γ‐T. Postprandial hyperglycemia decreased FMD (p<0.05) in both groups. Postprandial FMD area under the curve (AUC) increased in both groups following γ‐T supplementation consistent with greater baseline FMD. Baseline nitro‐γ‐T and total cysteine and postprandial malondialdehyde AUC were lower in all men following γ‐T whereas myeloperoxidase and C‐reactive protein were unchanged. Thus, short‐term γ‐T supplementation likely improves VEF by decreasing oxidative and nitrative damage, but without affecting insulin resistance or inflammatory responses. Support provided by International Life Sciences Institute to RSB.Grant Funding Source: International Life Sciences Institute
A Randomized Trial of Multivitamin Supplementation in Children with Tuberculosis in Tanzania
Authors: Mehta Saurabh , Mugusi Ferdinand M , Bosch Ronald J , Aboud Said , Chatterjee Anirban , Finkelstein Julia L , Fataki Maulidi , Kisenge Rodrick , Fawzi Wafaie W 
Published: June 2021

BackgroundMultivitamin supplementation may enhance the health of children with tuberculosis (TB); however, its efficacy as an intervention has not been examined.Methods255 children with TB were randomized to receive either a daily multivitamin supplement or placebo in the first eight weeks of anti‐TB therapy. They were followed up for the duration of supplementation through clinic and home visits to assess anthropometric indices and laboratory parameters, including hemoglobin and albumin.ResultsMultivitamin supplementation resulted in a significant improvement in hemoglobin levels at the end of follow‐up; the median increase in children receiving multivitamins was 1.0 g/dL, compared to 0.4 g/dL in children who were given placebo (p<0.01). No significant differences were observed in growth indices, including height and weight gain. Among children who were also co‐infected with HIV (n=87; 34%), daily multivitamins had a smaller but statistically significant increase in albumin levels compared with placebo.ConclusionsMultivitamin supplementation for eight weeks improved the hematological profile of children with TB. Larger studies with a longer period of supplementation are needed to confirm these findings and assess the effect of multivitamins on clinical outcomes including treatment success and growth failure.ClinicalTrials.gov Identifier: NCT00145184Grant Funding Source: NIH 3U01AI045441
Zinc absorption from traditional meals by vitamin A‐depleted Bangladeshi women is greater in those with altered intestinal permeability, while iron absorption is not affected
Authors: Perez‐Exposito Ana Beatriz , Woodhouse Leslie , Haskell Marjorie J , Jamil Kazi M , Barfod Gry , Wahed Mohammed A , Brown Kenneth H 
Published: June 2021

BackgroundZinc and iron deficiencies have been associated with altered intestinal mucosal function.ObjectivesTo examine the relationship between intestinal permeability and zinc and iron absorption in non‐pregnant, vitamin A‐depleted Bangladeshi women (plasma retinol < 1.05 umol/L).MethodsZinc absorption from a traditional test meal was assessed using the dual stable isotope tracer ratio method, and iron absorption was assessed by erythrocyte incorporation of a stable iron isotope 10 days after the oral dose. Intestinal permeability was evaluated with the lactulose‐mannitol (L:M) test.ResultsFractional zinc absorption (FZA) was greater in women with altered intestinal permeability (urinary L:M recovery ratio >0.07) than in women with L:M ≤0.07 [FZA = 36.1 ± 9.4 % vs. 24.9 ± 10.3 %, respectively, p = 0.038]. Iron absorption from meals did not differ by intestinal permeability status [2.9 % (1.4, 5.8) vs 2.6 % (1.8, 3.8) p = 0.79]. Zinc absorption was marginally correlated with the urinary lactulose:creatinine ratio (p = 0.07).ConclusionsZinc absorption from a traditional meal in vitamin A‐depleted Bangladeshi women is greater in those with altered intestinal permeability than in those with normal permeability, but iron absorption is not affectedFunding was provided by HarvestPlus. Ana Perez‐Exposito was partially funded by the International Nutrition Foundation/Ellison Medical Foundation.
Bioconversion of B‐cryptoxanthin into polar metabolites other than vitamin A
Authors: Mein Jonathan Ryan , Dolnikowski Gregory , Russell Robert , Wang Xiang‐Dong 
Published: June 2021

Several well‐implemented cohort studies have shown dietary intake or blood levels of β‐cryptoxanthin, but not other pro‐vitamin A carotenoids such as β‐carotene and α‐carotene, to be strongly associated with a reduced risk of lung cancer. This has raised an important question as to whether the potential protective activity of β‐cryptoxanthin, a mono‐hydroxy carotenoid, is due to the intact molecule or formation of unique metabolites. The objective of the present study was to investigate the excentric cleavage activity of mammalian carotene‐9′,10′‐oxygenase (CMO2) towards β‐cryptoxanthin. We found that baculovirus‐generated recombinant ferret CMO2 cleaved β‐cryptoxanthin at both the 9,10 and 9′10′ double bond in a protein‐, time‐ and dose‐dependent manner. In addition to the expected apo‐10′‐carotenal and β‐ionone cleavage products, HPLC and LC‐MS analysis identified the formation of two unique metabolites: 3‐OH‐apo‐10′‐carotenal and 3‐OH‐β‐ionone. Furthermore, in the presence of NAD+, incubation of 3‐OH‐apo‐10′‐carotenal with ferret liver S9 lysate resulted in the dose‐dependent formation of 3‐OH‐apo‐10′‐carotenoic acid. In conclusion, the bioconversion of β‐cryptoxanthin by CMO2 into 3‐OH‐apo‐10′‐carotenoids is significant because 3‐OH‐apo‐carotenoids may represent biologically important metabolites with specific biological activities related to human health.Grant Funding Source: R01CA104932
Tomato carotenoid bioaccumulation and gene expression of carotenoid cleavage enzymes are altered by soy germ in male Copenhagen rat tissues
Authors: Zuniga Krystle Erin , Erdman John W 
Published: June 2021

Epidemiological studies have supported the hypothesis that dietary intake of soy foods or tomato products is associated with a lower risk of prostate cancer. However, little is known about how simultaneous consumption may impact absorption, tissue distribution, and metabolism of tomato carotenoids. Diets containing 10% tomato powder (TP) (n=17), 2% soy germ (SG) (n=16), neither (n=17), or a combination (TP+SG) (n=13) were fed to 12 week old male Copenhagen rats for approximately 24 weeks. Bioaccumulation of tomato carotenoids were lower in various tissues of rats fed TP + SG compared to TP alone, especially in androgen sensitive tissues. Testes, seminal vesicles, and ventral prostate of the TP+SG group had lower levels of lycopene (p=0.001, p=0.04, and p=0.002, respectively) and β‐carotene (p=.003, p=.003, p=.001, respectively) than the TP group. Gene expression of carotenoid 15,15′‐monooxygenase (CMOI) and carotenoid 9′10′‐monooxygenase II (CMOII) were measured in the liver, duodenum mucosa, and prostate. Rats fed TP had significantly higher hepatic CMOI expression compared to the control group (p=.046), but expression was not significantly different from rats fed SG or TP+SG. Liver expression of CMOII was not significantly different between groups. Our results suggest that dietary soy germ may alter uptake and/or metabolism of carotenoids. Research supported in part by USDA NNF 2005‐03750.Grant Funding Source: NIH