bluuebunny/crossref_metadata_2025 huggingface dataset:
Nuclear Pore Complex [NPC] and Epigenetic Regulator Proteins Share Conserved Domains for Interaction with Functional Non‐Coding ncRNA
Authors: Wissler Joerg E.
Published: June 2021

OBJECTIVE

Functional small hairpin ncRNA may address defined homologous helix‐nucleating domains shared in epigenetic regulator proteins entangled in growth, metabolic syndromes, vascular, cancer, epi‐ and genetic information indexing of the epigenome. Whether they may also participate in nucleocytoplasmic transport was investigated by searching conserved interaction domains in nuclear [epigenome] and NPC proteins [Mol. Biol. Cell 20 Suppl, 534, 2009; Ann. N.Y. Acad. Sci. 1137: 316–342, 2008].

RESULTS

Edited, modified, redox‐ and metalloregulated functional small hairpin ncRNA [<200n] as bioaptamers in RNP complexes were obtained from transcriptional RNA output of cells activated by extrinsic [environmental] factors. Several components of chromatin formation modulation and NPC were found having conserved homologous helix‐nucleating structural [proteomic] domains in terms of K/RxxxH [K/R3H], i.e.‐t/s/xK/R/q/n/hxxxH/y/n/q/e/d/r/kx7‐9h/xx7–9h/xx5–20K/R/q/n/e/h‐with accessory canonical basic [R/K]n, R/K‐zipper, SR/K/RS, EF‐hand and/or HxxxH/y/n/q segments. They include nucleosomal assembly, histone deacetylase, DNA‐C5‐methyltransferase, sirtuin, hypoxia‐inducible factor, nuclear pore complex, nucleoporin, importin, exportin and nuclear transport factor protein families. By redox‐ and metalloregulated multivalent interaction with molecular imprints in protein components, such ncRNA may display functions in NPC, its transfer or shuttle factors in synergism with indexing of the nuclear epigenome.

CONCLUSIONS

The results suggest that in the “terra incognita” of molecular dynamics of NPC and orchestration with genomic and nucleocytoplasmic processes, new functions associated with epigenetic imprinting and inheritance await discovery.
The novel gene C17orf37 is regulated at both transcriptional and post‐transcriptional levels
Authors: Rajendiran Smrithi , Dasgupta Subhamoy , Vishwanatha Jamboor
Published: June 2021

C17orf37, a novel gene located in human chromosome 17q12, is abundantly expressed in cancer cells compared to minimal expression in normal cells. We have previously shown that C17orf37 is a membrane bound molecule triggering downstream signaling through the AKT/NF‐κB pathway, up‐regulating migration and invasion of cancer cells. The reasons for aberrant increased expression of C17orf37 in cancer cells are still unclear. The objective of this study is to identify deregulated mechanisms leading to differential regulation between normal and cancer cells. Our data indicate that C17orf37 mRNA and protein expression is induced in DU145 and PC3 cells upon treatment with interleukin (IL) 4 and 13. IL‐4 and IL‐13 mediated signaling pathways have been shown to be commonly activated in prostate cancer patients. C17orf37 is also post‐transcriptionally regulated by specific microRNAs (miRNAs) which are lost in cancer cells. miRNA expression profiling pattern in normal and prostate cancer cells indicate several miRNAs that can directly bind to 3′UTR of C17orf37 mRNA. Knockdown of riboendonuclease III family proteins ‐ Drosha and Dicer show an increase in C17orf37 expression compared to mock controls. Identification of specific miRNAs resulting in differential expression of C17orf37 is currently in progress. Targeting C17orf37 may lead to novel therapeutic intervention for cancer patients. (NIH‐ CA109593 & MD001633)
Poly(A)‐specific ribonuclease (PARN): Molecular mechanisms of mRNA cap and poly(A) tail recognition
Authors: Virtanen Anders , Henriksson Niklas , Nilsson Per , Wu Mousheng , Song Haiwei
Published: June 2021

PARN is a homodimeric, processive, and cap‐interacting 3′ exoribonuclease that degrades mRNA poly(A) tails. To reveal mechanisms by which PARN recognizes cap and poly(A) we have performed a structural and functional study of PARN. A crystal structure and subsequent mutational analysis revealed that amino acid residues involved in cap recognition were crucial for cap‐stimulated deadenylation activity and poly(A) hydrolysis. A detailed kinetic analysis showed that PARN harbors specificity for adenosine recognition in its active site. We propose that two binding pockets, which interact with the nucleotides surrounding the scissile bond, play a pivotal role in providing specificity towards adenosine recognition. Interestingly, the binding pocket required for recognition of the penultimate adenosine residue is also required for cap binding and efficient hydrolysis. We conclude that the cap‐binding site partially overlaps with the active site of PARN.

This work was supported by the Swedish Research Council, the Linneus Support from the Swedish Research Council to the Uppsala RNA Research Centre and the Lennanders Foundation at Uppsala University. H.S. acknowledges financial support from the Biomedical Research Council of A*STAR (Agency for Science, Technology, and Research) in Singapore.
Systematic studies on the recognition of 3′‐end methylated small silencing RNAs by PAZ domains
Authors: Ding Lei , Huang Ying , Tian Yuan , Patel Dinshaw J. , Ma Jin‐Biao
Published: June 2021

The PAZ domain is a conserved RNA‐binding module that is only found in two core components of RNA silencing, Argonaute and Dicer. Previous studies showed that the PAZ domain specifically recognizes 3′‐end 2‐nucleotide overhang of small RNAs. Recently, it has been demonstrated that many small silencing RNAs, such as miRNAs and siRNAs in plants, piRNAs in animals and siRNAs in Drosophila, are methylated on the 2′‐OH of the 3′ terminal nucleotide. This kind of modification resulted in a reduction of binding affinity between the small RNAs and the PAZ domain from human AGO1. In order to further understand the function of the PAZ domain from different Argonaute proteins, systematic binding and structural studies of PAZ domains derived from two subfamilies of Argonaute proteins (AGO and PIWI) and four species (human, fly, fish and worm) are carried out. Results from different binding assays (fluorescence anisotropy and isothermal titration calorimetry) indicate that PAZ domains from human and Drosophila AGO1 proteins bind to 3′‐end methylated small RNAs with reduced binding affinities but PAZ domains from Drosophila AGO2 and other PIWI subfamily Argonaute proteins bind to 3′‐end methylated small RNAs with enhanced binding affinities, which consist with deeper and wider pockets found in PAZ domains from Drosophila AGO2 and PIWI subfamily proteins. Structural comparison implies that PAZ domains from divergent Argonaute proteins may have distinct functions.
How HIV‐1 Rev Gets its Groove On
Authors:   Galileo SMART team, Colton Shannon , Herman Tim , Giles Kurt , Koo Ben , Jeske Sabine , Fernandes Jason , DeBella Stan
Published: June 2021

HIV‐1 Rev is an essential viral regulatory protein. Rev is responsible for nuclear export of partially and unspliced viral RNAs; these viral RNAs are required for the production of essential late‐stage viral proteins such as Gag and Env, and also serve as genomic RNAs for virion packaging. Rev's structure is key to understanding its function. Rev has several major domains, including an arginine rich RNA binding domain. This domain forms a marginally stable alpha helix that binds a major groove in the viral RNA. We have built physical 3D models of Rev as well as a portion of the HIV‐1 RNA that Rev recognizes, the Rev Response Element (RRE). These models allow us to visually illustrate the details of this important RNA‐protein interaction. A better understanding this interaction could allow for novel anti‐HIV therapies that target this essential interaction.

The SMART (Students Modeling A Research Topic) program was established by the Center of BioMolecular Modeling at the Milwaukee School of Engineering and involves partnerships between university researchers and local high schools. This work is funded by NIH‐NCRR‐SEPA and HHMI.
Modification of the pH‐dependent function of the HMPV F protein
Authors: Hackett Brent , Dutch Rebecca
Published: June 2021

Paramyxoviruses are a diverse class of negative‐strand RNA viruses that contain numerous clinically relevant human pathogens, including human metapneumovirus (HMPV), a recently identified respiratory pathogen for which there is currently no treatment. Amidst the open reading frames encoded within the HMPV genome is the fusion (F) protein, an integral membrane protein responsible for fusion of the viral and host membranes, resulting in delivery of a viral particle into the host cell and subsequent infection. In this study, we sought to further examine the novel pH‐dependent mechanism of the HMPV F protein recently described in our lab. Prior studies of other paramyxoviral fusion proteins have identified moieties in the HRA and F2 regions of the protein that interact in the pre‐fusion state to form a putative trigger for membrane fusion. Site‐directed mutagenesis of key residues in these regions of HMPV F resulted in dramatic effects fusogenic activity in vitro, including effects on the dependency of low pH. We hypothesize that these mutations may have effects on F pre‐fusion stability and conformational change to the post‐fusion state. The data suggest that alanine mutations of some of these residues may lower stability of the pre‐fusion state, leading to increased triggering, and in some cases, may be sufficient to overcome the requirement for low pH for transition to the post‐fusion conformation. Research supported by NIH R01 AI051517.
Association Between 5‐HT Receptor Gene Promoter Polymorphism and Obesity
Authors: Hemimi Neveen Salah El Din , El Salam Mona Mohamed Abd , Khalil Marwa Mahmoud , Ahmad Heba Mohamed Galal El Din
Published: June 2021

Background

Serotonin has an integral role in appetite control, sexual and social behaviour, and stress response and these all form part of the individual susceptibility to obesity. AA genotype of 5‐HT2A gene promoter polymorphism was associated with increase receptor expression and binding.

Aim

To study the relation between the G1438A polymorphism of the 5‐HT2A gene and the susceptibility to obesity.

Material& Method

69 individuals with BMI < 25 Kg/m2 and 120 individuals with BMI ≥ 25 Kg/m2 were enrolled in the study. The G1438A polymorphism of the 5‐HT2A gene was detected by restriction fragment length polymorphism polymerase chain reaction (RFLP‐PCR) of peripheral blood DNA samples. Analysis of data was done by using SPSS program.

Results

Genotype frequencies for AA and GA& GG were 25% and 75% respectively in the group with BMI ≥ 25 Kg/m2 compared to 52.2% and 47.8% in the normal BMI group. Carriers of GA& GG genotype were at a significant high risk of developing obesity when compared with carrier of AA genotype (OR=3.3, 95%CI= 1.7–6.1). Moreover, the mean energy intake per day was significantly increased among carriers of GA&GG genotype when compared to AA genotype (p< 0.05).

Conclusion

G allele was a risk factor for obesity.

Source of Fund: Ain Shams University.
Adipose tissue responses to binge and restrictive eating in ground squirrels
Authors: Kohlnhofer Bridget M , Cooper Scott T , Maher Margaret A
Published: June 2021

Fat cells provide peripheral feedback to the brain for appetite regulation. Adiponectin, leptin, and resistin are adipocyte hormones and PPARs are diet (fatty acid) sensitive transcription factors that modulate glucose and fatty acid metabolism. The 13‐lined ground squirrel (Spermophilus tridecemlineatus) is a natural model of binge and restrictive eating associated with hibernation and coincident alterations in metabolism. We have compared key regulators of metabolism and appetite, in adipose tissue from non‐hibernating versus hibernating ground squirrels, with quantitative PCR. Amplification crossing points were determined for each gene and normalized to that of actin. Preliminary studies revealed adiponectin and PPAR alpha transcription were up‐regulated 6 and 16‐fold, respectively, while PPAR gamma transcription was down‐regulated by 3‐fold during hibernation. Adiponectin results are consistent with observations by others that levels of this hormone are inversely related to fat mass, which would be predicted to decline during hibernation. Future studies will address dietary versus endogenous nutrient contributions to these observations. Funded by University of Wisconsin‐System and WiSys grants.
Rapamycin‐sensitive Co‐localization of mTOR to the Mitotic Spindle
Authors: Shenberger Jeffrey S , Zhang Lianqin
Published: June 2021

Protein synthesis and mTOR (mammalian target of rapamycin) activity are suppressed during mitosis. Recently, phosphorylated mTOR has been localized to the mitotic spindle. To investigate functions of mTOR during mitosis, we examined the impact of rapamycin (RAP) on mTOR localization and mTOR complex (mTORC1/2) assembly. Using immunofluorescence we observed that mTOR co‐localizes to mitotic spindle fibers and the mid‐body in normal, transformed, and cancer cells. Incubation of HeLa cells with mTOR activity‐suppressing concentrations of RAP (0.1–3 μM) for 30 min failed to alter the localization of mTOR. In A549 cells, 0.1 μM RAP dispersed mTOR from spindle fibers. Pre‐treatment with the kinesin inhibitor monastrol (ML) prior to RAP stabilized mTOR on monopolar asters in A549 cells. Immunoprecipitation of mTORC1/2 revealed similar raptor/mTOR/rictor interactions in mitotic and interphase HeLa and A549 cells. In both cell types, RAP reduced raptor in mTORC1 from mitotic and interphase fractions even in the presence of ML, while the content of rictor in mTORC2 was unaffected by RAP or ML. Our findings reveal that the association of mTOR with the mitotic spindle in A549 cells is not exclusively mediated by raptor interactions, though these interactions are sensitive to RAP. Differences in the avidity of mTOR for the spindle fibers may have implications for the cell‐specific anti‐proliferative effects of RAP.
Mechanisms of Mitochondrial Dysfunction Induced by Sulfur Mustard in Human Epidermal Keratinocytes (HEK)
Authors: Martens Margaret E
Published: June 2021

Exposure of HEK to the chemical warfare agent sulfur mustard (SM) significantly disrupts cell energy metabolism. Our data suggest that one or more enzymes of respiratory complex I may be directly inhibited by SM and that complexes II, III and IV are relatively insensitive. To further localize sites of inhibition, we examined the partial reactions NADH‐ferricyanide reductase and cytochrome c oxidase. 100 μM SM did not inhibit either reaction at 24 hrs post‐exposure. 500 μM inhibited cyt. c oxidase by 25%, possibly secondary to cell death. We conclude that SM inhibits anterior to cytochrome c for succinate and between NADH dehydrogenasecyt. c for NAD‐linked substrates. Our previous data also suggest that SM causes irreversible mitochondrial damage parallel to late stage (18–24 hrs) losses in ATP and cell viability. We examined mitochondrial membrane permeability with JC‐1 and calcein‐AM+CoCl2. At ≤ 8 hrs we saw no changes with ≤ 500 μM SM. At 18–24 hours, 100 μM SM had no effect, but 300 and 500 μM caused losses in membrane potential that, in some cells, coincided with abnormal morphology. The data suggest that collapse of the mitochondrial membrane potential is a key event in SM‐induced cell death, perhaps at the level of the transition pore. Supported by DTRA‐JSTO, Med. S&T Div. The opinions, interpretations, conclusions, and recommendations are those of the author and are not necessarily endorsed by the US Army or the DoD.
Self‐Sufficient Catalytic System of Cytochrome P450cam, Putidaredoxin, and Putidaredoxin Reductase Provides a Useful Cell‐based Enzymatic Reaction
Authors: Eun Chang‐Yong , Lim Young‐Ran , Han Songhee , Park Hyoung‐Goo , de Montellan Paul R. Ortiz , Kim Donghak
Published: June 2021

The catalytic turnover of cytochrome P450cam (P450cam) from Pseudomonas putida requires two auxiliary reduction partners, the iron‐sulfur protein putiaredoxin (Pd) and the flavoprotein putidaredoxin reductase (PdR). We report the functional expression in Escherichia coli of tricistronic constructs consisting of P450cam encoded by the first cistron and the auxiliary proteins, Pd and PdR by the second and the third. The expression level of P450cam was 500 nmol per liter of culture. Transformed bacterial whole cells efficiently oxidized (1R)‐(+)‐camphor, the normal substrate of the enzyme, to 5‐exohydroxycamphor. More interestingly, this whole cells system successfully oxygenated a monoterpene, limonene, and converted it to (−)‐perillyl alcohol, an anticarcinogenic compound with limited availability. These bioengineered E. coli cells expressing three proteins possess a heterologous self‐sufficient P450 catalytic system that may have advantages in terms of low cost and high yield for the production of the fine chemicals. This study was supported by a Korea Research Foundation Grant funded by the Korean Government (MOEHRD, Basic Research Promotion Fund)(KRF‐2008‐331‐E00030)
CyclinB/cdk1‐mediated phosphorylation of the PR‐Set7 histone methyltransferase is required for its nuclear localization and stability during mitosis
Authors: Wu Shumin , Rice Judd C.
Published: June 2021

We previously demonstrated that accumulation of PR‐Set7 and the histone modification that it creates, monomethyl‐H4K20, during G2/M are essential for mammalian cell cycle progression. However, it remains unclear how PR‐Set7 protein is regulated during mitosis. Based on recent findings, we hypothesized that PR‐Set7 is a mitotic phosphoprotein and that this phosphorylation event affects PR‐Set7 function during mitosis. Here we discovered that human PR‐Set7 is phosphorylated specifically by cyclinB/cdk1 and exclusively at G2/M. We found that PR‐Set7 phosphorylation occurs early during mitosis and that this event is required for the proper localization of PR‐Set7 within the nucleus. In addition, phosphorylation stabilizes PR‐Set7 protein levels by directly preventing its ubiquitination and proteasome‐mediated degradation. Importantly, impairment of PR‐Set7 function resulted in global chromatin decondensation, cell cycle and segregation defects. Collectively, our results reveal a novel mechanism by which cyclinB/cdk1‐mediated phosphorylation of PR‐Set7 functions to stabilize and direct this chromatin‐modifying enzyme to ensure normal cell division.
Mapping domains of anti‐apoptotic Bcl‐xL required for interaction with cell cycle check point regulator Aven
Authors: Datan Emmanuel , Roelofs Brian , Hardwick J. Marie
Published: June 2021

Members of the Bcl‐2 family of proteins, known for their critical apoptotic functions, have often been shown to influence cellular processes based on knock‐out or over expression studies. However, the outcomes of these approach do not show how Bcl‐2 family members actually bring about such phenotypes or outcomes in over expressing, under expressing or knock‐out conditions. This void left by knock‐out/down or over expression studies is the avenue of protein‐protein interaction research. Mapping and crystallography has been the usual approach in protein‐protein interaction studies. In this study the interaction of a member of the Bcl‐2 family protein Bcl‐xL with cell cycle check point regulator Aven was mapped though point mutations in the BH homology domains of Bcl‐xL. After sequence verification and subsequent expression of the mutant constructs/plasmids with HA‐Aven in Hela cells, HA tagged Aven was then co‐immunoprecipitated with other complexing proteins. The mutant Bcl‐xL that was bound to Aven was identified through Western blots. Data suggests that amino acids 2, 18, 25, 131, 135, 136, 137, 181 are important for Bcl‐xL interaction with Aven. These amino acids belong to the BH4, BH1 and BH2 domains of Bcl‐xL. In addition to mapping out the required amino acids for interaction, results also show that the kind of Bcl‐xL mutant and it's concentration may affect or be affected by the stability of Aven in vitro. The results of this study should be verified in other conditions especially were Bcl‐xL and Aven are critical proteins.
Unraveling Spiderman's glue silk
Authors: Vierra Craig Anthony , Hsia Yang , Geurts Paul , Zhao Liang , Tuton‐Blasingame Tiffany , Blasingame Eric , La Mattina Coby
Published: June 2021

Spiders spin high performance fibers that have diverse mechanical properties for movement, prey capture and protection of eggs. To investigate the mechanism by which spiders anchor their fibers to solid matrices, we solubilized the attachment discs from black widow spiders (cob weavers) and performed in‐solution tryptic digests followed by MS/MS analysis to identify novel peptides derived from glue silks. Combining matrix‐assisted laser desorption ionization tandem time‐of‐flight mass spectrometry and cDNA library screening, we have identified the first spider glue fibroin of the silk gene family called Pyriform Spidroin 1 (PySp1), as well as a closely related fibroin in orb weavers using conventional nucleic acid‐nucleic acid hybridization. Biophysical characterization of black widow spider attachment disc fibers using atomic force microscopy, as well as biochemical analyses of PySp1 using circular dichroism and NMR, demonstrate that PySp1 has distinct molecular features relative to other silk family members. Given the unique biophysical and chemical properties of PySp1, this fibroin could be ideal for use in spinning artificial silks through genetic engineering for different commercial applications, which include body armor, tissue engineering, sutures for the medical industry, and materials for the automobile and aerospace industry. This research was supported by the NSF RUI Grant MCB‐0544087.
Inherently Antibacterial Hydrogels: Altering Activity via Tryptophan/Arginine Interactions
Authors: Larsen Tyler John , Salick Daphne , Nagarkar Radhika , Schneider Joel
Published: June 2021

Hydrogels are heavily hydrated materials that show considerable promise as artificial extracellular matrices for use in tissue regenerative therapies. The development of antibacterial hydrogels has been of great interest to the hydrogel research community as a means to combat the threat of infection during material implantation. We have developed MAX1, a self‐assembling β‐hairpin peptide hydrogel whose surface exhibits inherent antibacterial activity against several pathogens prevalent in hospital settings. Under physiological conditions, MAX1 self‐assembles into a crosslinked, mechanically rigid hydrogel, but the process is too slow for use as an injectable implant. This study aims to design a hydrogel with rapid self‐assembly and high rigidity under physiological conditions while maintaining potent inherent antibacterial activity through incorporation of cation‐π interactions, which are common in antibacterial peptides found in nature. A new β‐hairpin peptide (RWMAX1) was designed, incorporating a cross‐strand R/W pair into the MAX1 sequence. The folding and self‐assembly properties were assessed using circular dichroism and rheology and the antibacterial activity was investigated against gram positive S. aureus and gram negative E. coli. RWMAX1 gels were found to possess both favorable physical and antimicrobial properties.
Characterizing Y532H disease‐causing mutation and N‐terminal domains of the human Wilson protein
Authors: MUIA JOSHUA , Huffman David , Banci Lucia , Bertini Ivano , Wang Shenlin
Published: June 2021

The human Wilson protein (ATP7B) is a copper transporting ATPase that is involved in copper homeostasis. Mutations in the gene coding for this protein leads to Wilson Disease (WD), a hepatological disorder characterized by impaired excretion of copper in bile. We characterized Y532H disease‐causing mutant, and its interaction with a copper chaperone, HAH1 (also known as Atox1). We examined structural effects of Y532H mutation in domains 5 and 6 wild type construct of ATP7B. It was found that the mutation alters the solubility of the expressed protein, decrease overall protein stability as observed by circular dichroism (CD) spectroscopy and does not interfere with copper binding. The 15N HSQC experiments by NMR reveal an essentially folded protein, a phenomenon also supported by CD experiments. The amide chemical shifts are highly similar between the wild type and mutant WLN5‐6 for the domain 6 (G80‐Q149), as shown by chemical shift variation (CSV) data, and the most affected amino residues are those spatially close to mutation which includes Q4‐F7, K47‐H(Y)48 (mutation) and V52‐I53. In conclusion, the Y532H mutation is perceived to perturb the hydrogen bond network in domain 5, which is critical for a compact structure, thus inducing motions on the side chains and break the linkage between two domains, causing inter‐domain motion and structural heterogeneity, which is absent in the wild type. (This work was supported by Wilson's disease Association grant)
ANTIBACTERIAL ACTIVITY OF Co(II) COMPLEXES WITH SOME HETEROCYCLIC COMPOUNDS
Authors: Govori Sevdije R , Ajvazi Njomza , Buza Aida , Haziri Arben , Rifati Arleta
Published: June 2021

The antibacterial activity of cobalt (II) complexes with some heterocyclic derivatives were investigated in vitro against Staphylococcus aureus, 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 (1 – 5) in concentration 1mg/l, 0.5mg/l and 0.1mg/l.

Comparing the inhibitory activities of the tested complexes, it was found that these complexes were more active than heterocyclic compounds (A – E).
Synthesis of Totarol Analogs for Inhibition of FtsZ
Authors: Rutaganira Florentine Uwamahoro , Kim Michelle B , Shaw Jared T
Published: June 2021

The recent emergence in antibiotic‐resistant strains of pathogenic bacteria has driven research on the discovery of new molecular scaffolds with efficient synthesis. Natural product synthesis provides a method for scaffold discovery. More than two‐thirds of clinically used antibiotics are natural products or their semisynthetic derivatives. Totarol, a diterpene natural product, is an antibacterial substance effective against gram‐positive bacteria (1.5 μM in B. subtilis). Although totarol may have multiple targets, experiments confirm totarol's ability to inhibit FtsZ protein, a prokaryotic analog of tubulin essential for bacterial cytokinesis. Since the total synthesis of totarol is known, we propose the synthesis of totarol analogs to gain insight into structure‐activity relationships between totarol and FtsZ. We utilized a copper‐catalyzed coupling of a benzylic grigard to an epoxyacetate. However, synthesis of the benzyl grignard proved to be difficult. Consequently, a second route was proposed that uses concepts from the umplong reaction. The analysis of chemical structures focuses on the use of Gas Chromatography – Mass Spectrometry and 1H NMR.
Using Naturally Derived Compounds to Combat HIV: A Case for Complementary and Alternative Medicinal Modalities
Authors: Narayan Vivek , Chiaro Christopher , Henderson Andrew J , Prabhu K. Sandeep
Published: June 2021

Anti‐retroviral therapies for AIDS have typically targeted HIV proteins integrase, protease, and reverse transcriptase. Transactivator of transcription (Tat), an early HIV protein that increases the efficiency of proviral transcription, and replication, is now being looked at as a possible new target for anti‐HIV therapies. We have previously shown that electrophilic prostaglandin, 15d‐PGJ2, is capable of modifying the Cys rich region in Tat irreversibly such that it loses its transactivating property. In an effort to identify more such molecules, which can inhibit Tat, we have explored a selected few naturally derived compounds with electrophilic carbon centers. Of the several compounds tested, celastrol, a triterpene extracted from Tripterygium wilfordii, inhibited Tat‐dependent HIV transcription and replication at low concentrations, and that this inhibition was independent of the NF‐kappa B pathway. Using a biotinylated derivative of celastrol, we show that the Cys‐thiols in Tat are targeted in a covalent Michael addition reaction. Furthermore, circular dichroism spectroscopy was used to demonstrate changes in the secondary structure of Tat upon interaction with celastrol. Thus, a structural perturbation of Tat could likely impact the processivity of RNA polymerase II, leading to the inhibition of transcription. These data suggest that redox‐modulating natural compounds such as celastrol have the potential to be developed as anti‐retroviral therapeutics for HIVAIDS.
Effect of Phospholipids on the Stability and Absorption of Açaí Anthocyanins
Authors: Cardona Jorge Alfredo , Talcott Stephen T , Mertens‐Talcott Susanne U
Published: June 2021

The purposes of this study were to determine the efficacy of phospholipids as aiding agents in the absorption of anthocyanins isolated from açaí on a monolayer from human Caco‐2 colon cancer cells and to evaluate their effect on anthocyanin stability. Transepithelial electrical resistance (TEER) was monitored to check cell confluence and integrity. Cells remained in culture for 18–21 days to guarantee enterocytic cell differentiation and P‐gp expression. Anthocyanin content was determined spectrophotometrically and by HPLC‐MS analysis. Açaí contained two anthocyanins, cyanindin‐3‐glucoside (C3G) and cyanidin‐3‐rutinoside (C3R). C3G and C3R alone were poorly absorbed (1.6% and 1.7% respectively). The presence of phospholipids (5000 mg/kg) significantly increased the absorption of C3G and C3R (5.66% and 10.2% respectively). Color degradation followed first‐order kinetics. Anthocyanin degradation was similar within treatments at every temperature illustrating that the presence of phospholipids did not affect anthocyanins degradation kinetics. In contrast, effects of temperature were evident. Higher temperatures resulted in higher losses of anthocyanins and reduced shelf life of the samples. Refrigeration (5 °C) resulted in an average half life of 97.7 days whereas higher temperatures (25 °C and 37 °C) resulted in reductions of 43% and 59% in half life respectively.

Grant Funding Source: ASN
Antioxidants induced cytokine alterations indicative of a better prognosis after ACL surgery
Authors: Barker Tyler , Martins Thomas B. , Hill Harry R. , Kjeldsberg Carl R. , Trawick Roy H. , Leonard Scott W. , Walker James A. , Traber Maret G.
Published: June 2021

Following anterior cruciate ligament (ACL) surgery, muscle weakness is commonly observed and may be potentiated by pro‐inflammatory cytokines. Because vitamins E and C (AO) reportedly modulate cytokines, the purpose of this study was to evaluate the influence of AO supplements on cytokines pre‐ and post‐ACL surgery. A randomized, double‐blind, placebo‐controlled study was conducted in men undergoing ACL surgery, who were supplemented with AO or placebos (PL; n = 10/group). Supplementation began ~2‐w prior to and concluded 3‐mo after surgery. Fasting plasma AO and cytokines were measured. IL‐6, IL‐10, and IL‐2rα increased (P < 0.05) and PGF2α decreased (P < 0.05) following ACL surgery and during NSAID medication. AO supplementation ameliorated (P < 0.05) the increase in IL‐10. Thus, despite standard of care NSAID medication, a systemic cytokine response occurred post‐surgery, and AO protected against the increase in IL‐10. By 3‐mo post‐surgery and after the discontinuation of NSAIDs, IL‐1β increased (P < 0.05), but for a given increase in IL‐1β, AO potentiated the increase (P < 0.05) in IL‐4. The AO potentiated increase in IL‐4 may limit the systemic release of IL‐1β and protect against the adverse sequelae of inflammatory responses. In conclusion, AO supplementation induced immunological alterations after ACL surgery indicative of a better prognosis.

Source of support: USANA Health Sciences, Inc., SLC, UT.
Association of dietary and serum folate in a cohort of asymptomatic HIV+ patients in Botswana
Authors: Marlink Richard , Bussmann Hermann , Masmouie Hoda , Burns Patricia J , Tsalaile Lesedi , Raesi Mpho , Makhema Joseph , Campa Adriana , Baum Marianna K
Published: June 2021

Objective

Deficiency of folate is common in HIV infection and is associated with increased risk for cardiovascular disease, hypertension and anemia. We determined the variables associated with risk for folate deficiency in HIV infected antiretroviral naïve HIV infected adults in Botswana.

Methods

After consenting, 140 HIV+ adults were administered medical, physical activity and 24‐hour recall questionnaires. Clinical exams were performed. Blood was drawn for CD4 cell count, viral load and serum folate levels.

Results

The mean age was 34.0±8.21, 48% (n=67) were male and 15.2% had low serum folate (<5 ng/mL). Serum folate was significantly correlated with physical activity (r=0.244, p=0.011), BMI (r=−0.255, p=0.004), and female gender (r=−0.560, p<0.001). CD4 cell count and viral load were not significantly related to serum folate. Total folate intake was predictive of serum folate levels (β= 2.476, p=0.015) in a dose‐response relationship after controlling for physical activity, smoking, hypertension and gender.

Conclusion

Folate intake is reflected in serum levels in a dose response manner and is a good marker of folate intake from diet and from supplements in HIV+ adults. Serum folate levels may be used as an index of adherence in micronutrient supplementation trials that contain folate.

Funded by NIDA
Dietary vitamin D intake and food sources of US and Canadian population
Authors: Jonnalagadda Satya S , Albertson Ann M. , Joshi Nandan , Moore Kathy , Sharma Bhavna , Culp Julie M.
Published: June 2021

Objective was to determine vitamin D intake and food sources in US (n=7837) and Canadian (n=4025) populations using 7‐ or 14‐d food intake data from a nationally representative population. Mean vitamin D intake ranged from 3.7 to 6.0 mcg/d. One‐third of the 2–50 y age groups met their AI for vitamin D, except US women, 19–50 y, where only 23% met their AI. Less than 10% of individuals over the age of 50 y met their AI for vitamin D. The top 10 food sources of vitamin D in the diets of U.S. participants were milk, meat, fish, eggs, ready‐to‐eat (RTE) cereal, pasta and rice, dairy and non‐dairy frozen desserts, poultry, shellfish, and vegetable recipes; among the Canadian participants they were milk, meat, fish, margarine, eggs, egg recipes, pasta and rice, dairy and non‐dairy frozen desserts, vegetable recipes, and soups. Participants who frequently consumed RTE cereal had significantly higher total vitamin D intake vs. those who did not consume or were infrequent consumers. Frequent breakfast consumers had significantly higher vitamin D intake vs. those who did not consume breakfast or were infrequent consumers. Given the inadequate intake of vitamin D, emphasis needs to be put on increasing dietary sources of vitamin D, including vitamin D fortified foods, to help individuals meet the vitamin D dietary recommendations.

Funding: General Mills Bell Institute of Health and Nutrition, Minneapolis, MN
Low dietary intake of vitamina B12 predicts hyperhomocysteinemia in Mexican women.
Authors: Anaya‐Loyola Miriam Aracely , Alvarez Carina Sosa , Loria Jorge Luis Rosado , Allen Lindsay H
Published: June 2021

The objective of this cross‐sectional study was to evaluate the impact of diet on homocysteine (Hcy), vitamin B12 and folate status in women living in Queretaro (central Mexico). Average mean values for age, weight, height and BMI were 37.6±10.3 ys, 68.4 ± 14.6 kg, and 151.6±5.2 cm and 29.8±6.0 kg/m2, respectively. Mean concentrations of serum vitamin B12, folate and Hcy were 331.5±209.9 pmol/L, 21.1±8.2 nmol/L and 10.9±3.2 nmol/L. Vitamin B12 deficiency was observed in 11.4% of the participants. In addition 21.2% presented marginal status. Only 5.4% of the women were folate deficient. Prevalence of hyperhomocysteinemia was 32.6%. B12 deficient women had inadequate intakes of vitamin B12 in about 50% (<EAR). Increase in Hcy was related to dietary intake and serum concentrations of vitamin B12. Being vitamin B12 deficient increase 2.8 times the risk of presenting hyperhomocysteinemia. Dietary folate was no related to Hcy, but serum values were. Vitamin B12 deficiency as a result of low dietary intake was the most important factor associated to high Hcy in blood in these Mexican women.
Mapping of anemia prevalence among Honduran children ages 6–60 months using global positioning system data
Authors: Stoltenburg Ashley Ann , Kemmer Teresa M. , Gidvani‐Diaz Vinod , Lougee Douglas , Coello Miguel , Amador Wilmer E. , Lynch Julia , Thanapura Pravara
Published: June 2021

Based on national data, 1/3rd of all Honduran children under age 5 suffer from anemia. No studies to date have mapped the prevalence of anemia within rural Honduras. Objectives were to 1) determine and map the prevalence of anemia within children ages 6 to 60 months and 2) determine socioeconomic, growth, nutrition, and health factors associated with anemia. Randomized household surveys were conducted in collaboration with the Honduran Ministry of Health. Hemoglobin was obtained from 851 children living within 16 health centers. Altitude, latitude, and longitude were obtained at each household using a handheld global positioning system. Anemia was defined as hemoglobin < 11.0 g/dL (altitude adjusted). Overall prevalence of anemia was 29.8%. Anemia prevalence in one health center was <20%, 20–30% in 7 health centers, 31–40% in 6 health centers, and 40–50% in 2 health centers. Mapping data shows the prevalence of anemia in each health center. Risk factors associated with anemia included living at elevations >5,000 feet, (p=.012), mother was anemic (p=.005), weight‐for‐age z‐scores < −2 (p=.049), clinic distance > 1hour from home (p=.004), consumed meat < 2 times per month (p=.029), > 7 people living in household (p=.019), house constructed from material other than brick (p=.030), mother attended school < 3 years (p=.015), and child breastfed > 24 months (p=.000). By targeting health centers at highest risk for anemia, the Honduran Ministry of Heath can use this data to implement prevention measures as a means of reducing childhood morbidity. Supported by Henry M. Jackson Foundation for Advancement of Military Medicine.
Reductions of atherogenic lipoproteins are related to concentrations of plasma carotenoids in women following a Mediterranean‐style low glycemic diet
Authors: Barona Jacqueline , Kopec Rachel , Jones Jenifer , Lerman Robert , Schwartz Steven , Fernandez Maria Luz
Published: June 2021

We investigated the effects of a Mediterranean‐style low glycemic diet with (n=20) or without (n=15) a medical food containing soy protein and plant sterols, rho iso‐alpha acids, and proanthocyanidins on plasma carotenoid concentrations and on lipoprotein subfractions and size in 35 women classified with metabolic syndrome. Independent of treatment, women had an increase in plasma lutein (P <0.0001), zeaxanthin (P < 0.05) and beta‐carotene (P < 0.0001), while lycopene was reduced (P < 0.05) after 12 wk. LDL cholesterol (LDL‐C) was reduced from 139.5 ± 34.6 to 113.4 ± 33.7 mg/dL (P < 0.0001). In addition, decreases were observed in the atherogenic subfractions: large VLDL (P < 0.05), small LDL (P < 0.00001) and medium HDL (p < 0.05). Plasma lutein concentrations at wk 12 were negatively correlated with large VLDL (r= −0.349, P < 0.05) and small LDL (r= −0.340, P < 0.05) and positively correlated with LDL size (r = 0.337, P < 0.05). In contrast, plasma lycopene was positively correlated with LDL‐C. The data indicate that women complied with the dietary regimen by increasing vegetable intake. The decrease in high glycemic foods rich in lycopene such as pasta and pizza may be responsible for the lowering of this carotenoid in plasma after 12 wk. The results also suggest that lutein transport in plasma is associated with the formation of larger LDL while the lowering in plasma LDL‐C is associated with the lowering of plasma lycopene. [Supported by MetaProteomics LLC, Gig Harbor, WA]
The dose‐response effects of the amount of oil in salad dressing on the bioavailability of carotenoids and fat‐soluble vitamins in salad vegetables
Authors: Agustiana Agatha , Zhou Yang , Flendrig Leonard , White Wendy S.
Published: June 2021

The objectives were to define the dose‐response relation of the amount of added oil and: 1) the absorption of carotenoids, phylloquinone and tocopherols in salad vegetables; 2) the absorption of retinyl palmitate formed from the ingested provitamin A carotenoids, α‐ and β‐carotene. Women (n = 12) each consumed 5 salads containing equivalent amounts of carrot, cherry tomato, romaine lettuce and spinach. The salads with salad dressings containing 0, 2, 4, 8 or 32 g tocopherol‐stripped soybean oil were ingested in random order separated by ≥ 2 weeks. Blood samples were collected at baseline and 2, 3.5, 5, 7, and 9.5 h postprandially. Chylomicron fractions were extracted and analyzed by HPLC with coulometric array electrochemical detection. When the salads were ingested with 0 g oil, there was negligible absorption of α‐ and β‐carotenes, lutein, lycopene, phylloquinone, retinyl palmitate, α‐ and γ‐tocopherols. For α‐ and β‐carotenes, lycopene, retinyl palmitate, and α‐ and γ‐tocopherols, absorption was increased with each amount of oil compared with 0 g oil (P < 0.05). The absorption of each carotenoid and fat‐soluble vitamin was highest with 32 g ingested oil (P < 0.002). Supported by Unilever.
Dietary fats with increased ratio of unsaturated to saturated fatty acids enhance absorption of carotenoid and vitamin E by increasing both efficiency of micellarization and lipoprotein secretion.
Authors: Chitchumroonchokchai Chureeporn , Failla Mark L. , Ferruzzi Mario G. , Campbell Wayne W.
Published: June 2021

Although it is well established that dietary fat promotes the bioavailability of carotenoids and other dietary lipophiles, insights regarding the effects of different lipid sources on processes associated with absorption of these plant compounds remain unclear. We previously reported that triacylglycerides (TG) with higher ratios of unsaturated to saturated fatty acids (FA) increased the efficiency of micellarization of carotenes. Here we examined the influence of FA composition of TG on uptake of β‐carotene (BC), lutein (Lut) and αTC and their basolateral secretion in chylomicrons by Caco‐2 cells. Differentiated Caco‐2 cells were pretreated 4h/d for 4d by apical exposure of cultures to mixtures of saturated, mono‐unsaturated and poly‐unsaturated FA similar to those present in butter (76:24:0) and olive(12:74:14), canola(9:56:35) and soybean oil (16:24:60). On the following day, micelles containing βC, Lut and αTC were included along with the FA mixtures in apical medium. Uptake of test compounds was unaffected by micelle FA composition, whereas micelles rich in unsaturated FA strongly promoted basolateral secretion of BC, Lut and αTC. The number and size of chylomicrons (total TG and TG to apoB48/100 ratio) also were significantly increased by chronic exposure to micelles with FA composition mimicking vegetable oils. Therefore, it appears that ingestion of TG enriched in with unsaturated FA enhances bioavailability of carotenoids and αTC by increasing micellarization during digestion and transepithelial transport by absorptive epithelial cells. Supported by USDA NRI Grant # 2007‐02313 and OARDC.
Genotype and diet alter carotenoid bioaccumulation and the expression of carotenoid cleavage enzymes in CMO‐I KO, CMO‐II KO, and wild‐type mice
Authors: Ford Nikki Ann , Clinton Steven K. , von Lintig Johannes , Erdman John W.
Published: June 2021

Two primary enzymes are involved in carotenoid metabolism, carotenoid monooxygenase I (CMO‐I) and carotenoid monooxygenase II (CMO‐II). Evidence suggests β‐carotene is centrally cleaved by CMO‐I to form vitamin A. We hypothesize that acyclic carotenoids, like lycopene, are eccentrically cleaved by CMO‐II. We investigated carotenoid bioaccumulation in adult male wild‐type (WT) mice or mice lacking either CMO‐I or CMO‐II. Mice consumed a control AIN‐93G diet or AIN‐93G diets with 10% tomato powder, 100mg lycopene/kg diet (DSM beadlets) or placebo beadlets for 4 or 30 days. Lycopene accumulated in CMO‐II KO mouse tissues while β‐carotene accumulated in CMO‐I KO mouse tissues thus supporting our hypothesis. RNA expression of CMO‐I and CMO‐II in response to diet and/or genotype was evaluated by RT‐PCR (ANOVA with Tukey's post hoc analysis). We observed tissue specific expression patterns that show significant interactions between diet and genotype. For example, tomato powder feeding decreased CMO‐II expression in livers of WT mice, but increased expression in CMO‐I KO mice. Interestingly, CMO‐II, but not CMO‐I, showed detectable expression in the prostate and was significantly lower in CMO‐I KO mice compared to WT mice. Overall, diet and genotype alter tissue expression of key carotenoid cleavage enzymes. Supported by NIH grant PHS‐1‐RO1 CA125384.

Grant Funding Source: NIH grant PHS‐1‐RO1 CA125384
Dietary cotton‐seed oil attenuates secretion of adhesion molecules by Human Umbilical Vein Endothelial Cells (HUVEC)
Authors: Sundaresan Sinju , Sodhani Shradha , Vijayagopal Parakat , Mills Nathaniel , Imrhan Victorine , Prasad Chandan
Published: June 2021

We investigated the effect of post‐prandial HDL isolated from healthy females consuming a single high‐fat meal rich in cotton seed oil (CSO) on TNF‐α stimulated soluble Intercellular Adhesion Molecule (sICAM‐1) and Vascular Cell Adhesion Molecule (VCAM‐1) expressions in HUVEC. In a randomized cross‐over design, eleven healthy females consumed 1 of 2 iso‐caloric meals containing either CSO or blended palm oil (BPO) (1 g of fat/kg BW). Blood was drawn at 0, 3 and 6 hrs after meal and HDL isolated by sequential centrifugation. HUVEC incubated with TNF‐α were treated with HDL (2, 4, 8 μg/mL) and protein expression of adhesion molecules secreted were determined by Western blotting. Serum TNF‐α, oxidized LDL, sICAM‐1, and sVCAM‐1 were determined by ELISA. One‐way repeated ANOVA was performed to determine differences between three time periods and two groups. HDL isolated from CSO but not BPO attenuated expression of sICAM‐1 and VCAM‐1, in a dose‐dependent manner. Levels of oxidized LDL in the CSO were significantly lower than the BPO (p<0.05). The CSO‐mediated attenuation of adhesion molecule expression should reduce the adherence of inflammatory cells to the vascular endothelium, thus reducing the risk for cardiovascular disease.

Supported by Texas Department of Agriculture
Pharmacokinetics of dietary equol in ovariectomized rats
Authors: Legette LeeCole L , Prasain Jeevan , Arabshahi Alireza , Barnes Stephen , Weaver Connie M
Published: June 2021

Recent findings indicate that soy isoflavones and their metabolites may play a major role in mitigating post‐menopausal bone loss. Equol, a metabolite of soy isoflavone daidzein produced by intestinal bacteria, has shown some potential but only 30–50% of US population is capable of converting dietary daidzein to equol. There is limited data on the pharmacokinetics of dietary equol and its metabolites. This study was conducted to assess the levels of equol and its conjugates in plasma for a 24h period resulting from oral administration of dietary equol in ovariectomized Sprague Dawley rats. Plasma samples were analyzed for using liquid chromatography‐tandem mass spectrometry. Although the majority of equol metabolites present were glucronidated conjugates (≥99%), there were low levels of sulfated conjugates present, specifically equol monosulfate. Pharmacokinetic parameters for equol monosulfate were: Cmax (21±7 nmol/L), Tmax (2.8±2h), AUC0‐t (237±79 nmol h/L), AUC0‐inf (600±390 nmol h/L), T1/2 (32±24h). Pharmacokinetic parameters for glucronidated equol were: Cmax (10.5±2.9μmol/L), Tmax (0.88±0.75h), AUC0‐t (11±14 μmol h/L), AUC0‐inf (179±14 μmol h/L), and T1/2 (16±3h).

Grant Funding Source: NIH