bluuebunny/crossref_metadata_2025 huggingface dataset:
The impact of fish‐oil fatty acids on postprandial vascular reactivity (959.22)
Authors: McManus Sean , Vauzour David , Cassidy Aedin , Minihane Anne Marie
Published: June 2021

Vascular dysfunction is a significant cardiovascular disease (CVD) risk factor. High fat meals tend to incur a postprandial loss of vascular reactivity, with our previous work showing that the inclusion of fish oil, providing the long chain n‐3 PUFA, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), in part ameliorated the response. In this randomised placebo controlled crossover study (n=25), the individual impact of EPA versus DHA (4.2g) on postprandial vascular reactivity in males aged 35‐55 years old at 1.5 relative risk of CVD was assessed, along with underlying physiological mechanisms.

Both EPA and DHA improved the postprandial vascular augmentation index by 16.7% and 15.8% respectively, with the effect for DHA but not for EPA reaching significance. In addition to the vascular measures, a range of biochemical markers of vascular function and lipidomic analyses will be presented. The trial advances currently understanding of the individual vascular benefits of EPA vs. DHA and the relative impact of these fatty acids on overall cardiovascular health.
The prevalence of micronutrient deficiency among women (959.3)
Authors: Abduldayeva Aigul , Smagulova Indira , Doszhanova Gulnur
Published: June 2021

Goal ‐ is based on the studying the prevalence of micronutrient deficiencies amongchildren and women of reproductive age to develop evidence‐based recommendations aimed at improving the effectiveness of programs to prevent micronutrient deficiency.

Subject: In our study we used a representative, random sample, carried out with the cluster method in the precinct of the principle areas of medical care for children 5 years of old. If the site has at least 60 children under 5 years of old, each second child was sampled, and if more than 60 children ‐ each third child (first child selected by random sampling). The total number of investigated persons was within 80‐86 women of reproductive age and children ‐ within 80‐92 people.

Results: The studies found that the average prevalence of anemia among children aged 6‐59 months was 35.2%, with the most susceptible to iron deficiency anemia in infants aged 6‐23 months (53.3%). The prevalence of anemia among non‐pregnant women was 39.0% among pregnant women ‐ 43.8%.

In children, the prevalence of folate deficiency was the highest (27.6%). Among non‐pregnant women frequent prevalence of folic acid deficiency was 37.0%.

The prevalence of vitamin A deficiency was higher among children living in Astana (37.4%) compared with the medium‐republican level (23.2%).
Effect of fasting on the neuronal Fto expression and intracellular localization in the rat brain (959.6)
Authors: Vujovic Predrag , Stamenkovic Stefan , Jasnic Nebojsa , Lakic Iva , Djordjevic Jelena
Published: June 2021

Fat mass and obesity associated protein (Fto) is a nucleic acid demethylase which exhibits preference for thymine and uracil. This suggests that methylated single‐stranded RNA, rather than DNA, may be the primary Fto substrate. Fto is expressed in myriad of tissues, but the expression is especially high in the hypothalamic regions regulating energy homeostasis.

Since Fto expression is bidirectionally regulated, as a function of nutritional status, our aim was to examine the effect of 48 h fasting on the Fto expression in various brain regions.

qPCR and Western blot were used to assess the Fto mRNA and protein expression respectively. Immunofluorescence was employed to address Fto intracellular localisation.

Fasting upregulated overall hypothalamic Fto mRNA and protein expression. More importantly, it was documented for the first time that majority of the enzyme was localized outside cell nuclei in some neurons of paraventricular, ventromedial nucleus and lateral hypothalamic area. Change in the Fto intracellular localization was not observed in neurons of arcuate nucleus, cortex and hippocampus.

Although fasting increased hypothalamic Fto content, it did not universally affect Fto expression and intracellular localization in this brain region.

Grant Funding Source: Supported by the Serbian Ministry of Education, Science and Technological Development Grant No 17302
Sphingosine 1‐phosphate signaling in vascular and immune systems (96.2)
Authors: Hla Timothy
Published: June 2021

Sphingosine 1‐phosphate (S1P) is a simple lysophospholipid produced from the metabolism of sphingomyelin. Although originally thought as an intracellular second messenger, work over the past two decades has shown that S1P acts primarily as an extracellular lipid mediator. In addition, due to its central position in sphingolipid metabolism, it plays an important role in intracellular lipid homeostasis. Vertebrates evolved to use S1P as an extracellular mediator since their genomes contain S1P transporters and receptors. The first S1P receptor was cloned as an orphan G protein‐coupled receptor (GPCR) from vascular endothelial cells in 1990 and de‐orphaned in 1998. We now know that five distinct GPCRs (S1P1‐5) that are widely expressed mediate most of the actions of this lysophospholipid mediator. Some of the well‐studied actions of S1P include its essential roles in vascular development and immune cell trafficking. Inhibition of GPCR‐dependent S1P signaling results in therapeutic efficacy in the autoimmune disease of remitting, relapsing multiple sclerosis (RRMS). Indeed, the first S1P receptor inhibitory drug, Fingolimod, is now approved as an oral medication to treat RRMS in over 40 countries. This recent development has caused a flurry of activity to develop novel S1P receptor‐based therapeutics .

This talk will discuss cellular sources of plasma S1P, biological roles of HDL‐bound Apolipoprotein M, the major plasma S1P chaperone and the role of S1P receptor‐1 in vascular and immune homeostasis and ontogeny.

Better understanding of fundamental principles involved in S1P signaling is anticipated to help in the development of novel therapeutic approaches to control many human diseases.
The role of I614 and E615 in unnatural substrate recognition in Taq DNA polymerase mutants (967.5)
Authors: Gochi Andrea , Schultz Hayley , Leconte Aaron
Published: June 2021

DNA polymerases enable key DNA biotechnologies by performing enzymatic amplification of DNA with high efficiency and fidelity, enabling DNA's use in polymerase chain reaction (PCR), sequencing techniques and many different applications in therapeutics and diagnostics. While these astounding properties of DNA polymerases enable the use of these technologies, they also restrict them because they do not recognize modified substrates. Three previous studies have evolved DNA pol I from Thermus aquaticus (Taq) to recognize 2’ modifications of DNA; while these have resulted in an increase in modified substrate recognition, these enzymes are not active enough for practical use. Here, we describe efforts to identify the key mutations that impart unnatural activity on these previously identified enzymes. In particular, we have focused on mutations at I614 and E615 since they occur in all three previously identified mutants. Data show that much of the activity that is present in the previously evolved enzymes is largely a result of the E615G mutation. Mutations at I614 have been found to have no effect when present alone, but have also been found to have varying effects on enzymatic activity when in conjunction with the E615 mutation. Current work is also being done to further investigate the synergistic interactions of mutations of I614 and E615G. These efforts will elucidate key features of the evolution of unnatural substrate recognition by mutant Taq enzymes and will aid in future polymerase engineering efforts.
Antioxidant and antiradical activity of essential oils of Kazakhstan plants (972.1)
Authors: Kazbekova A.T. , Moldabekov K.Zh. , Seitembetov T.S. , Zeinuldina A.S. , Dalenov E.
Published: June 2021

One of the current interests in the finding of potentially active natural substances is the matter of methodology for evaluation of biological activity. In modern medicine plant extracts are widely used, particularly those showing antioxidant and antiradical effects. Therefore determination of antiradical activity of individual compounds isolated from plants, the same as aggregated extracts. That can be investigated further with possible correlations between these parameters.

The main purpose of the work was the in vitro study of antioxidant and antiradical activity of essential oils of Lophanthus schrenkii, Achillea cartilaginea and luteolin glycoside (Y.Suleimen, Eurasian National University, Kazakhstan).

Comparative in vitro study of the antioxidant and antiradical activity of essential oils of Lophanthus schrenkii, Achillea cartilaginea and luteolin glycoside using the method of inhibition of DPPH by standard substance butylated hydroxyanisole (BHA). It has high antiradical activity and can be compared with investigating sample. We have determined high antiradical activity of BHA, which is higher than the activity of luteolin glycoside, essential oils of Lophanthus schrenkii and Achillea cartilaginea, i.e. the antiradiacla activity of investigated samples can be arranges in the next order BHA luteolin glycoside Achillea cartilaginea Lophanthus schrenkii.
Lingonberry (Vaccinium vitis‐idaea) anthocyanins protect cardiomyoblasts against ischemia‐reperfusion injury (973.1)
Authors: Isaak Cara , O Karmin , Siow Yaw L
Published: June 2021

Lingonberry contains more antioxidants and anthocyanins than most berries and consumption of these phytochemicals has been shown epidemiologically to reduce risk of cardiovascular disease. Since the mechanisms of protection against cellular injury require elucidation, this study examined the effects of complete lingonberry, anthocyanin‐enriched, and phenolic‐enriched extracts on apoptosis in H9c2 rat cardiomyoblasts during simulated ischemia‐reperfusion (IR). This model induced a 20‐fold increase in nuclei condensation and JNK phosphorylation and a 2‐fold increase in caspase‐3 activation. Despite having low antioxidant capacity, the anthocyanin‐enriched extract and pure anthocyanins protected cells against apoptosis by significantly inhibiting IR‐induced nuclei condensation, caspase‐3 activation, and JNK phosphorylation to the same degree as the complete extract. These results indicate that anthocyanins may exert anti‐apoptotic effects via mechanisms unrelated to their antioxidant capacity. This research highlights the effects of dietary anthocyanins on cell signaling in IR injury and bolsters the body of evidence for the health benefits of anthocyanin‐rich foods. (AAFC Project ID 1129)

Grant Funding Source: Supported by AAFC
Cardiac glycosides with selectivity for the α2 isoform of NaK‐ ATPase (974.1)
Authors: Katz Adriana , Tal Daniel , Karlish Steven
Published: June 2021

Cardiac glycosides (CG) are specific inhibitors of Na,K‐ATPase. Na,K‐ATPase consists of, α and β subunits and an FXYD regulatory protein. There are four isoforms of α and three isoforms of β, which are expressed in a tissue‐specific fashion. α1 is the general purpose isoform, α2 is expressed in heart, and plays a key role in cardiac contraction and thought to be an important target of CG’s. Inhibitors of α2 could induce cardiac contraction (positive inotropy) with minimal toxic (arhythmogenic) effects. α2 is also expressed in the eye, in non‐pigmented cells of the ciliary epithelium, and is the motor for production of aqueous humour. The aim of this project was to develop CG’s with selectivity for human α2 isoforms of NaK‐ATPase that could effectively control raised intra‐ocular pressure (IOP), and induce cardiac contraction with minimal toxic effects.

We combined techniques from organic synthesis, biochemistry‐protein expression, purification, binding assays and ATPase inhibition measurements. We used recombinant purified human α1β1, α2β1 isoforms to screen a large No.of known CG’s, and found that the sugar moiety determines isoform selectivity. Based on these finding we synthesized a series of perhydro‐1,4‐oxazepine derivatives of digoxin, modified in the third digitoxose moiety. Several derivatives show improved α2/α1 selectivity, close to 8‐fold. The ability of these derivatives to prevent an acute pharmacologically‐induced increase in IOP in rabbit eyes has been studied. Two α2‐selective derivatives efficiently prevent the ocular hypertension, by comparison with known CG's. These selective inhibitors of α2 isoform can turn into safer treatment of heart failure and high ocular pressure, the main risk factor in glaucoma.
Reversal of multidrug resistance by novel inhibitors of human P‐glycoprotein in cell culture (974.2)
Authors: Follit Courtney , Brewer Fran , Vogel Pia , Wise John
Published: June 2021

Multidrug resistance (MDR) and chemotherapy failure are often due to overexpression of members of the ABC‐transporter superfamily of proteins like MDR1 P‐glycoprotein (P‐gp). These membrane transporters protect cells from the accumulation of harmful toxins by using ATP hydrolysis to move a broad range of compounds across the plasma membrane. In MDR, however, the administration of chemotherapeutics selects for cancerous cells which overexpress P‐gp thereby creating a population of cancer cells resistant to many types of chemotherapeutics. Using targeted molecular dynamic simulations and massively parallel drug docking studies, we have identified several novel compounds that inhibit transport by P‐gp by targeting the ATP binding domains. Biochemical and biophysical studies have shown that these inhibitors do indeed bind to the nucleotide binding domains of P‐gp and inhibit ATP hydrolysis. In the studies reported here, we have been able to re‐sensitize MDR prostate and ovarian cancer cell lines in culture to multiple chemotherapeutics when treated with the novel compounds. The compounds reverse the MDR phenotype to sensitivities similar to the original, non‐MDR parental cell lines. Limited toxicity was observed when non‐cancerous cells were treated with the P‐gp inhibitors alone. The results suggest that these novel P‐gp inhibitors may serve as good drug leads for treating MDR and chemotherapy failure.

Grant Funding Source: Supported by NIH/NIGMS 1R15GM094771 ‐ 01A1 to PDV and JGW and the Communities Foundation of Texas
Inhibition of energy transduction in P‐glycoprotein (974.3)
Authors: Brewer Fran , Follit Courtney , Rossi Isolina , Wheelis Christopher , Wise John , Vogel Pia
Published: June 2021

Cancer chemotherapy failures often involve the over‐expression of ABC‐transporters like the MDR1 P‐glycoprotein (P‐gp). These transporters normally protect the cell by pumping toxins and xenobiotics across the plasma membrane. Selection for cells over‐expressing P‐gp during chemotherapy, however, results in populations that are drug resistant. Recently, we used massively parallel in silico drug docking to identify molecules predicted to interact with higher affinity at the energy transducing structures of P‐gp than at the drug transporting structures. In vitro assays verified that the compounds inhibited verapamil‐stimulated ATP hydrolysis and only marginally stimulated basal ATP hydrolysis activities. This indicates that these compounds do not interact with drug binding domains. The effects of the identified compounds on ATP binding was then studied using spin‐labeled ATP analogs and ESR spectroscopy. Results showed that some of the identified compounds decreased ATP binding stoichiometry to P‐gp while others decreased ATP binding affinities. These studies further showed that inhibition by the compounds was reversible. The investigations suggest that the discovered P‐gp inhibitors may make good drug leads. Experiments using ESR spectroscopy and site‐specific spin‐labeling of the drug binding domain are currently underway that may shed further light on the molecular mechanism of inhibition.

Grant Funding Source: NIGMS1R15GM094771 ‐ 01A1, Communities Foundations of Texas
Tetraiodothyroacetic acid (tetrac) receptor on integrin αvβ3: tetrac blocks activation of the integrin response to tumor cell irradiation (974.6)
Authors: Leith John , Hercbergs Aleck , Lin Hung‐Yun , Mousa Shaker , Tang Heng‐Yuan , Davis Paul
Published: June 2021

We have described a thyroid hormone‐tetrac receptor on plasma membrane integrin αvβ3 that mediates hormone analogue regulation of cancer cell proliferation and of tumor‐related angiogenesis (PJ Davis et al., Annu Rev Pharmacol Toxicol 51:99‐115, 2011). Liganded to this receptor, tetrac and nanoparticulate tetract (Nanotetrac) radiosensitize cancer cells (AH Hercbergs et al., Cell Cycle 10[2]:352‐357, 2011). One mechanism of radiosensitization is impairment of cellular repair of DNA double‐stranded breaks. We have defined in human prostate cancer (PC‐3) cells grown on the chick chorioallantoic membrane (CAM) the response of αvβ3 to ionizing radiation in the absence and presence of tetrac and Nanotetrac. PC‐3 grafts grow spherically in the CAM system and develop a core of hypoxic cells. Lysates of untreated PC‐3 cells contain about 6,900 αvβ3 molecules/cell (DuoSet R&D Systems) and ionizing radiation with 1‐10 Gy increased the number of αvβ3 moles/cell approximately 5‐fold over this radiation dose range. Investigated for state of activation (open conformation) by flow cytometry, the integrin was indeed activated 7‐fold over the 1‐10 Gy radiation dose exposure range, a protective response. Treatment of cells with tetrac or Nanotetrac (1‐2.5 μg/CAM) prevented the increase in cell number of αvβ3 molecules in response to radiation and blocked radiation‐induced activation of the integrin. Thus, radiosensitization of tumor cells in the CAM model that is conferred by tetrac formulations is complex. Inhibition by tetrac and Nanotetrac of the activation of integrin αvβ3 leads, via intracellular signaling mechanisms, to disordered repair in the nucleus of double‐stranded DNA breaks.
Regulation of ecto‐NTPDase I gene expression in Trypanosoma cruzi (974.7)
Authors: Gomes Natalia , Vidal Vitor , Moreira Otacilio
Published: June 2021

The ecto‐NTPDase I is an apyrase located on the outer surface of Trypanosoma cruzi plasma membrane, and previous data suggest that it possess a role to the infectivity and virulence of the parasite. In this study, we evaluate the mRNA levels for the Ecto‐NTPDase I by RT‐qPCR in distinct T. cruzi strains/clones and distinct evolutive forms. The Real Time PCR assays were performed with primers designed to amplify a 111bp sequence in the ecto‐NTPDase I T. cruzi gene and, as endogenous controls, a 268bp sequence in the calmodulin T. cruzi gene and a 100bp sequence in the GAPDH T.cruzi gene were used. We observed that the strains/ clones Cl‐Brener, Dm28c, 4167, LL014, 3663 and Y presented mRNA expression levels, respectively, 8,21±1,62; 7,17±1,51; 1,26±0,18; 1,20±0; 1,06±0,04; 0,94±0,08, times higher than the CL‐14 strain. Surprisingly, we observed that the epimastigote and amastigote presented expression levels, respectively, 14,35±2,04 and 10,23±1,16 times higher than the trypomastigote form. Moreover, following the parasite growth curve, we also observed that the expression of ecto‐NTPDase I raises in a time‐dependent manner, which could be related to a parasite response to a purine starvation in the cultivation medium. So far, our results suggest that the ecto‐NTPDase I expression is regulated at mRNA level, which is distinct between T. cruzi strains and evolutive forms.

Grant Funding Source: Supported by CNPq and FAPERJ
Human cathelicidin hCAP‐18/LL‐37 expression and subcellular localization in LNCaP and PC3 cell line models of prostate cancer: effects on growth, migration, and invasion (978.1)
Authors: Stone Sophia , Craig Maria , Deeble Paul
Published: June 2021

Human cathelicidin hCAP‐18 is an antimicrobial protein that is proteolytically processed to its active form, LL‐37, an alpha‐helical immune defense peptide. LL‐37 binds extracellular DNA and enhances cellular uptake of bacterial DNA containing unmethylated CpG motifs to activate the innate immune response. hCAP‐18 overexpression has been associated with disease progression in multiple epithelial cancers, and proliferative and pro‐metastatic effects of LL‐37 on cancer cells may contribute to a more aggressive tumor phenotype. We used RT‐PCR and western blot assays to characterize hCAP‐18 expression in LNCaP (androgen‐responsive) and PC3 (androgen‐independent) prostate cancer cell lines. We detect hCAP‐18 mRNA expression by RT‐PCR in both LNCaP and PC3 cell lines and observe higher expression in PC3 than in LNCaP cells using western blotting. Internalization and subcellular localization of LL‐37, alone and complexed with CpG oligonucleotides, were also investigated using immunofluorescence and western blot analyses. We find that LL‐37 appears to localize in endocytic vesicles and perinuclear regions of cells, and previous work by this group has shown cell line‐specific LL‐37 stimulation of growth (PC3), migration (PC3), and invasion (LNCaP). This work was supported by a Virginia Foundation of Independent Colleges Summer Undergraduate Research Fellowship.
All‐ambient processing of human blood samples: complete RNA and DNA stabilization with an automated workflow (981.10)
Authors: Liberal Vasco , Muller‐Cohn Judy , Muller Rolf , Whitney Scott
Published: June 2021

Gene expression profiles from blood samples are increasingly used to diagnose and monitor disease, requiring reliable preservation of RNA during sample collection and shipment. The highly dynamic transcription profiles can change rapidly after blood collection, potentially affecting interpretation of gene expression and ultimately dictate inadequate treatment. Biomatrica has developed a collection device for whole blood, which completely stabilizes the RNA in blood cells and, coupled with a robust RNA purification kit, provides a complete solution for all‐ambient blood sample processing. RNA isolation can be performed with multiple automated platforms, allowing an easy workflow and excellent results for low to high throughput sample processing.

For multiple studies, both DNA and RNA profiles from blood samples are needed, complicating logistics especially when large sample sets are required. Isolation of RNA and DNA from the same sample is ideal, since the use of separate samples could introduce operation errors as well as variation of the results. In order to accommodate the need for RNA and DNA isolations from the same blood sample stabilized in RNAgard Blood Tubes, we have developed an automated workflow for sequencial RNA and DNA purification, allowing for purification of both pathogen and host nucleic acids from the same stabilized blood sample.

Biomatrica’s ambient nucleic acid preservation technology, coupled with automated purification, can serve as the front end for research and molecular diagnostics, delivering accurate gene expression patterns and prove of extreme value for improved biomedical research and better patient treatment.
Metabolomic analysis of maternal energy metabolism (982.3)
Authors: Rajaee Arezoo , Murphy Aisling , Koos Brian
Published: June 2021

Objective Maternal O2 consumption progressively increases by about 20% over the course of gestation. This study was designed to test the hypothesis that the rise in maternal O2 consumption is reflected in quantitative perturbations of metabolites in critical pathways involved in maternal energy homeostasis.

Study Design Random maternal urine samples were obtained longitudinally from 16 normal gravidas with singleton pregnancies at 1) 6‐14 weeks' gestation, 24‐30 weeks' gestation, and 3) 6 weeks postpartum. The samples were analyzed on GC/MS and LC/MS/MS plateforms by Metabolon for 441 low‐molecular weight metabolites. The reported metabolites are those involved in glycolysis (glucose, lactate) and TCA cycle (alpha‐ketoglutarate, fumarate, malate, 3‐hydroxybutyrate). Concentrations were normalized to creatinine and expressed as scaled intensity. The log transformed data was analyzed by ANOVA with repeated measures to identify biochemicals that differed significantly between experimental groups. Data are expressed as fold‐changes between compared groups. Differences were significant at p<0.05.

Results Compared to postpartum (PP) values, urinary excretion of glucose and lactate was increased significantly by 26 ± 1.7 (SD) weeks' gestation (late pregnancy, LP) but not at 9 ± 2.3 weeks (early pregnancy, EP), as shown in Table 1. Malate, fumarate, 3‐hydroxybutyrate, and alpha‐ketoglutarate excretion also increased substantially with gestational age.

Conclusion The results are consistent with a substantial increase in maternal metabolic fluxes related to glycolysis and the TCA cycle.

BHBA, 3‐hydroxybutyrate. *p蠄0.05

Table 1. Fold‐changes for urinary metabolite ratios between groups.

Ratios
Glucose
Lactate
Fumarate
Malate
BHBA
Î±‐ketoglutarate

PP:EP
0.71
0.59
0.98
0.67*
0.81
1.71

PP:LP
0.05*
0.13*
0.49*
0.21*
0.26*
0.23*

LP:EP
14.13*
4.69*
2*
3.13*
3.16*
7.53*
Whole exome sequencing analysis of inter‐participant cortisol response: strategies for matching phenotype to genotype (983.1)
Authors: Murphy Patrick , Biette Kelly
Published: June 2021

The overarching goal of this project is to identify genetic variants that are indicative of an individual’s systemic and intracellular responses to corticosteroids. We previously observed inter‐participant variability in cortisol and nuclear hsp90 concentrations following exposure to mild cardiovascular stress, which could not be attributed to demographic or lifestyle characteristics or to single nucleotide polymorphisms (SNPs) in the glucocorticoid receptor, hsp90, or CRHR1 genes. We conducted whole exome sequencing of matched participant pairs who displayed similar demographic and lifestyle characteristics yet possessed distinct cortisol response phenotypes. Using an Agilent V4 exome enrichment platform and Illumina HiSeq2000 paired end (2 x 100 bp) sequencing, >44 million reads per sample were aligned to the hg19 reference build. This analysis resulted in average 30x coverage and combined analysis of >440,000 base pair variants. We developed a bioinformatics analysis pipeline to compare datasets of matched participant samples to one another and against the NHLBI dataset of >6,500 sequenced exomes. Filtering identified candidate causal variants associated with the phenotypic difference in cortisol response. These genes, which have the potential to impact corticosteroid‐mediated signaling and pharmacotherapy, are now being investigated for functional significance.

Grant Funding Source: Supported by NIH AREA grant GM086822
Genesis of antibiotic resistance (AR) I: monitoring of microbial pathogens with AR by Texas Clean Water Program in Rio Grande Basin including Amistad Reservoir (986.1)
Authors: Kannan Subburaj , Iracheta, Alyssa , Jimenez, Brenda , Lopez, Larizza , Martinez Karenny , Amezcua Judy , Subburaj Prabavathy
Published: June 2021

Water‐related (borne) diseases are categorized into four different classes: waterborne, water‐washed, water‐based, and water‐related insect vectors. The Pacific Institute Research Report (http://www.pacinst.org/) speculates that by 2020 a global mortality rate of 34‐76 million people will be observed due to water related diseases. One of the main sources for water in Southwest Texas is the Rio Grande River which receives its water influx from Las Vacas, Rio Conchos, Rio Rodrigo, and Rio Salado tributaries from Mexico. The Rio Grande River flows through the southwest border and is stored in the Amistad Reservoir located in Del Rio, Texas. The “Texas Clean Rivers Program” a collaborative program operated by The Amistad Reservoir ‐ International Boundary and Water Commission, US section(USIBWC) ‐ The Rio Grande Basin and The Texas Commission on Environmental Quality(TCEQ) monitor the microbial and water quality issues in Texas. As per the 2012 Basin Highlights Report for the Rio Grande Basin in Texas show that there were between 14,200 colonies/100 ml and 3,800,000 colony forming units/100ml of the fecal coli form bacteria in the water samples. High levels of Biological Oxygen Demand, and Chemical Oxygen Demand were also detected in the water samples tested in the urban areas of Del Rio, Eagle Pass and Laredo. Taken together the severity of the contamination, BOD and COD increases the mutagenic potential in the microbial pathogens (bacteria) to acquire antibiotic resistance through known and unknown mechanisms.

Grant Funding Source: PROFESSIONAL DEVELOPEMENT FUNDS ‐
Genesis of antibiotic resistance IX: monitoring of antibiotics, its metabolic break down intermediates by the United States ‐ Centers for Disease Control and Prevention (986.9)
Authors: Subburaj Subburaj , Dingler Margarita , Harbison Tiffany , Carrillo Esmeralda , Subburaj Prabavathy
Published: June 2021

CDC & P, a federal agency registry, formulating and advising the impact of the Antibiotics and Antimicrobial compounds, it’s precise role in tracking the use and its impact of the I. prescription, ii. Over the counter and iii. Non‐therapeutic use of Antibiotics is not well defined. CDC&P’s surveillance Systems in particular, The Emerging Infections Programs (EIP); Education Campaigns; and Antimicrobial Resistance, fall short of identifying, formulating, and providing specific advisory for each of the antibiotics either prescribed or antibiotics that are available over the counter (OTC) and non‐therapeutic use of the antibiotics. As per the advisory from CDC&P available on record, the CDC&P has neither role in monitoring neither antibiotics nor its metabolic breakdown products in potable water, sewage water, nor act as a registry for the harmful effects to the general public. The “Antibiotic Resistance Threats in the United States 2013” report published in April 2013 by EIP‐Active Bacterial Core Surveillance (ABCs) is based on the assumptions, expert opinions, and archived data. Though it is an update on prevailing trends in antibiotic resistance, recede of realistic advisory on the detection & preventive measure of on the antibiotics, and its metabolic breakdown products in the portable, untreated sewage, biosolids and processed clean water from raw sewage (domestic and medical/industrial sewage). Details on the dearth and desired aspect of the report will be presented in EB2014.

Grant Funding Source: PROFESSIONAL DEVELOPEMENT FUNDS ‐
Determination of Rac inhibitor Ehop‐016 in mouse plasma by ultra‐performance liquid chromatography tandem mass spectrometry (987.2)
Authors: Humphries‐Bickley Tessa , Corujo Francheska , Castillo‐Pichardo Linette , Hernandez‐O'Farrill Eliud , Vlaar Cornelis , Rodriguez‐Orengo Jose , Dharmawardhane Suranganie
Published: June 2021

The Rho GTPase Rac is an important regulator of cancer cell migration and cell invasion; processes required for metastatic progression. We previously characterized the small molecule Ehop‐016 as a novel Rac inhibitor in metastatic breast cancer cells in‐vitro (Montalvo‐Ortiz, et al., 2012). To investigate the efficacy of EHop‐016 in‐vivo, we used a mouse model of experimental metastasis, where mice with MDA‐MB‐435 mammary fat pad tumors were treated with intraperitoneal EHop‐016. At 25 and 40 mg/kg BW, EHop‐016 significantly inhibited mammary tumor growth and metastasis. In order to characterize the pharmacokinetics of Ehop‐016, we developed a rapid and sensitive method for the quantitation of Ehop‐016 in mouse plasma by ultra high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (UPLC/MS/MS). Plasma samples were pretreated with acetonitrile (ACN) to extract matrix proteins prior to injection into UPLC/MS/MS. Separation was carried out on an Agilent Poroshell 120 EC‐C18 column (3.0 x 50) mm using a mobile phase of 50% ACN/50% methanol/0.1% formic acid and 1mM ammonium fluoride. Ehop‐016 was identified from its accurate mass and retention times from the acquired full‐scan chromatogram and quantified by its peak areas. The linear range for the determination of analytes was 5 ‐ 1000 ng/mL. Ehop‐016 was quantified in samples from the in‐vivo study using this method.
New immunological assays for diagnosis of Schistosoma mansoni for clinical acute and/or chronic forms (990.1)
Authors: Grenfell Queiroz Rafaella , Silva‐Moraes Vanessa , Harn Donald , Coelho Paulo Marcos
Published: June 2021

Control constraints of schistosomiasis include the lack of diagnostic methods with high sensitivity. We develop a prospective study in southeast Brazil to standardize new sensitive diagnostic methods for Schistosoma mansoni infection. Currently, we are investigating 4 endemic areas together with 84 travelers infected in a freshwater pool. Sera, urine and saliva samples from all those patients has been used for the standardization of innovative methods, including acute, chronic, post‐treatment and myeloradiculopathy schistosomiasis patients. Comparisons are performed with eggs in feces, IgG titers, encephalomyelitis by NMR and clinical symptoms. We could diagnose acute patients 10 days post‐infection and also more than 95% of positive cases from chronic and low endemicity patients. New methods for IgG detection using purified glycoprotein or recombinant protein or peptides of 10 amino acids were superior to conventional ELISA. Best results were seen for recombinant protein with 100% of sensitivity. Finally, monoclonal antibodies were produced for improvement of the new methods by promoting direct diagnosis. Data showed 100% of sensitivity of chronic patients and 98% of acute patients. Financial support: Fapemig, CNPq, Capes, Fiocruz, Fiotec, PDTIS (Brazil). Fulbright, NIH, University of Georgia (USA).

Grant Funding Source: Fapemig, CNPq, Capes, Fiocruz, Fiotec, PDTIS (Brazil). Fulbright, NIH, University of Georgia (USA).
Osseous head in Canis canis (Bull‐dog): seriated CT‐scan and reconstructions (LB10)
Authors: LE FLOCH‐PRIGENT Patrice , VERDEILLE Stéphane
Published: June 2021

The osseous head of an adult bulldog (Canis canis) was CT‐scan in the frontal plane (Siemens, 64 bars). The cranium was laid on the table of examination with its longitudinal axis superimposed to the table's one; the orbito‐meatal (Frankfurt) plane was parallel to the table's horizontal plane. The head was 15,4 cm long. Several reconstructions were realized from the native cross‐sections with an average density. The cross‐sections in the three planes were then reconstructed with a high resolution, a thickness of 0,75 mm and an interval of 0,6 mm. 257 cross‐sections were obtained in the frontal plane, 132 in the para‐sagittal plane, and 158 in the horizontal plane. The cross‐sections were analyzed. The seriated CT‐scans of the entire and connected cranium and mandible in the dog in the three planes of the space brought a great accuracy in the analysis of the shape of the bones, of their achitecture of their volumes and of their relationships. This method is now unvaluable in the osseous morphological studies of the complex structures in every field of gross‐anatomy.
Food addiction in an urban sample of college students (LB117)
Authors: Dopler‐Nelson Mindy , Bullock Emily , Ackerson Leland
Published: June 2021

The objectives of this investigation are to create a shortened version of a survey to assess food addiction and to determine the prevalence of food addiction in an urban population of college students. Food addiction has a web of physiological causes very similar to other addictions that trigger the reward pathway and therefore influence inter‐individual variability in response to an obesogenic environment.

Methods: One question from each of the seven DSM‐IV‐TR substance dependence criteria were included in an online survey of interested college students at the beginning of the fall 2013 semester. Participants were from a convenience sample of students recruited by email invitation. Participants answered questions related to demographics and eating behavior to assess food addiction.

Results: 1980 students completed the online survey. 92.5% of the students answered at least five food dependence questions and were included in the data analysis. 46.2% had at least one symptom of food dependence. 9.1% of students were identified as having higher food addiction scores (3 or more symptoms) and 1.5% had clinical impairment to be classified with the diagnosis of food addiction.

Conclusions: These findings illustrate that nearly half of urban college students have symptoms of food addiction and approximately 10% meet the clinical diagnosis of food addiction. This may help to explain why certain individuals are predisposed to gain weight during their college years.

No external support was used to conduct this study.
Improving DNA extraction quality by the salting‐out method to prepare blood samples for real‐time PCR. (LB124)
Authors: Araujo Natalia , Carmona Bruno , Nogueira Glaucia Regina , Minicucci Marcor Ferreira , Conde Sandro
Published: June 2021

PCR, or “polymerase chain reaction”, is one of most useful techniques in genomics research. High quality DNA extraction is necessary to obtain satisfactory results. Some methods utilized to obtain DNA from blood include a simple salting‐out procedure for extracting DNA from human nucleated cells. This method is widely used by many laboratories because it can obtain DNA with sufficient quality to realize PCR, using materials that do not pose risk to researchers and is more cost‐efficient than other approaches. Although salting‐out has spread to many laboratories, its protocols do not highlight technical details of their steps that could affect final DNA quality and, for this reason, underwent some modifications in this work in order to obtain an efficient extraction. We used 2 to 4 mL of blood for each of 30 DNA samples. For the supernatant step in first EDTA wash, we established a subtraction pattern from 15 mL to 2.5 mL, independently of initial sample volume of blood, to avoid losing leucocytes. The number of washes required is directly proportional to loss of leukocytes in the blood sample, considering that 4 mL samples can receive five washes while 2 mL samples tolerate only four. It was noted that a “centrifuge brake” can reduce DNA extraction quality, thus indicating its inadvisability. All DNA samples were submitted to spectrophotometry, which showed the DNA/Protein ratio staying between 1.1 and 1.6. According to the literature, 93% of our samples were ready for real‐time PCR after modifications, indicating that those modifications were relevant for achieving efficient extraction. Integrity was confirmed by electrophoresis. All modifications made in the salting‐out method have improved the quality and integrity of DNA samples.

Grant Funding Source: FAPESP 2013/19808‐8
Bone Marrow ARA, EPA, and DHA are Correlated with Femur Minerals Content and Enzyme of Bone Formation in Growing Rabbits (LB152)
Authors: Alkhalifa abdulrahman , Nouri Doha
Published: June 2021

Bone Marrow ARA, EPA, and DHA are Correlated with Femur Minerals Content and Enzyme of Bone Formation in Growing Rabbits Alkhalifa,A.S, Al‐Nouri, D. M 1 Department of Food Sciences and Nutrition, Faculty of Food Sciences and Agriculture, King Saud University, Riyadh, Saudi Arabia ABSTRACT: The effects of long‐term supplementation with different dietary omega‐6/omega‐3 (ω‐6/ω‐3) polyunsaturated fatty acid (PUFAs) ratios on the bone marrow fatty acids level, plasma biomarkers of bone metabolism, and minerals content in bone were evaluated in rabbits. Weanling male and female New Zealand white rabbits were randomly assigned to five groups and fed ad libitum for 100 days on diets containing 70 g/kg different dietary oils which providing the following ω‐6/ω‐3 ratios: 8.7 g soy bean oil (SBO control), 21.8 g sesame oil (SO), 0.4 g fish oil (FO), 0.6 g DHA algae oil (DHA), and 0.7 g DHA and ARA algae oils (DHA/ARA). The bone marrow arachidonic (ARA), eicosapentaenoic (EPA), and docosahexaenoic (DHA) fatty acid levels were significantly influenced by and reflected the dietary ω‐6/ω‐3 ratios fed to rabbits. Rabbits fed on the FO diet maintained a lower ω‐6/ω‐3 ratio and a higher EPA and DHA levels , those fed on the DHA/ARA diet maintained a lower ω‐6/ω‐3 ratio and a higher ARA level, while those fed on the SO diet maintained a higher ω‐6/ω‐3 ratio and a lower ARA level. Plasma alkaline phosphatase (ALP) activity was significantly higher in male and female rabbits fed the DHA/ARA diet compared with those fed the control, SO, FO, or DHA diets. There was a significant main effect of dietary treatment on femur Ca, P, Mg, and Zn contents in both genders. This study confirmed that different dietary oil sources with varying ω‐6/ω‐3 ratios significantly altered the fatty acids level of bone marrow. In addition, the significant elevation in minerals content and the maintenance of optimal Ca/P ratio in bone of DHA/ARA and DHA fed groups beside the significant elevation in ALP activity in the DHA/ARA fed group proved that marine algae oils may be promising dietary sources for promoting bone mineralization and formation, thus improving bone mass during the growth stage.
Cadaveric study utilizing the ilizarov method for limb stabilization (LB17)
Authors: Summers Orianna , Welch Zachary , Lee Kelsey
Published: June 2021

By utilizing the Ilizarov principles as well as external fixation devices, the objective of this project is to actively demonstrate and analyze post‐fracture tibial‐fibular stabilization. This study will be conducted through the use of a cadaver supplied the University of California San Francisco’s Willed Body Program, and through the use of San Francisco State University’s Human Anatomy Laboratory. This cadaver will have stress applied to the lower limb in order to cause an accurate representation of a medically induced tibial‐fibular fracture. In order to correct this disjunction, the Ilizarov principles and external fixation device will be applied. Upon the placement of the Ilizarov external fixation device, the initial fracture will be corrected to provide stabilization of the tibia and fibula. This poster will aptly showcase the results of this study.

Grant Funding Source: None
Examining Irf4 genomic programming of lineage development using limited populations of purified immune cells via an optimized protocol for ChIP‐seq (LB176)
Authors: Giorda Kristina , Marcus Roland , Taylor Madison , Farias‐Hesson Eveline , Bluestein David , Ghosh Srinka , Meredith Gavin , Leach Sonia , O'Connor Brian
Published: June 2021

Dendritic cell (DC) lineages coordinate immune system activity through functional specialization. Irf4, a transcription factor, is required for CD11b+ DC lineage development from bone marrow stem cells and has been implicated in multiple inflammatory diseases such as asthma. The epigenetic consequences of immune specialization in CD11b+ DCs and relation to inflammatory diseases remain largely unexplored. One reason for this lack of genomic analysis stems from the difficulty of using highly purified, typically limited populations of cells in ChIP‐seq assays. A robust ChIP protocol was developed using limited amounts of K562 cells as a benchmark study. ChIP‐Seq analysis of the transcription factor CTCF or histone modifications strongly correlated with ENCODE data sets. This methodology was used to compare ChIP‐seq analysis of Irf4 genomic binding sites using flow sorted populations of 1, 3, 5, or 25 million CD11b+ lineage DCs. Very similar quality scores and quantitative results comparing Irf4 ChIP‐seq from 5 vs. 25 million sorted CD11b+ DCs were observed. Moreover, results indicate that fewer than 5 million flow sorted cells can be used to acquire high quality ChIP‐seq data. We identified genomic Irf4 binding sites proximal to genes, whose activity is consistent with CD11b+ DC lineage activity and/or known to contribute to inflammatory disease. To examine Irf4 functional regulation of these identified gene targets, RNA‐seq analysis with CD11b+ DCs and a related lineage, CD103+ DCs were performed. We integrated the expression analysis with the ChIP‐seq data and observed a unique CD11b+ DC gene expression program concordant with Irf4 loci association in comparison to CD103+ DC.
Age‐related differences in the plasma proteome in healthy adults: modulatory effect of regular aerobic exercise (LB178)
Authors: Santos‐Parker Jessica , Harrison Brooke , McQueen Matthew , Seals Douglas
Published: June 2021

Aging causes physiological dysfunction, some of which can be prevented or reduced by habitual aerobic exercise (AE). However, the molecular mechanisms underlying these effects remain largely unknown, particularly in humans. We used a novel high‐throughput molecular assay (SOMAscan v3.0, SomaLogic Inc., Boulder, CO) to characterize the effects of age and regular AE on the plasma proteome of healthy adult humans. From 1129 analytes, the expression of 48 proteins was greater and 37 lower in older (O; 64±6 yr; n=16) vs. young (Y; 24±4 yr; n=16) healthy men (false discovery rate [FDR] < 15%, p<0.01). Expression of the non‐collagenous extracellular matrix (ECM) protein dermatopontin was greater in O vs. Y (P=0.007), but was lower in older AE‐trained men (63±7 yr; n=8; VO2max = 49.6±3.7 mL/kg⋅min) compared with their sedentary counterparts (VO2max = 27.4±2.4 mL/kg⋅min) (FDR < 1.28%, p<0.001). Within O, no other group differences in plasma protein expression associated with exercise status met our FDR threshold (>35%). These preliminary results suggest that normal human aging is associated with several changes to the plasma proteome that may serve as molecular biomarkers of physiological dysfunction or disease risk. Regular AE status influences age‐related changes in circulating dermatopontin, a possible mediator of age‐related ECM remodeling.

Grant Funding Source: Supported by NIH AG013038 and RR025780
Autocrine hGH stimulated miRNA 96‐182‐183 cluster promotes epithelial‐mesenchymal transition and invasion in breast cancer (LB201)
Authors: Qian Pengxu , Zhang Weijie , Lobie Peter , Zhu Tao
Published: June 2021

Human growth hormone (hGH) plays critical roles in pubertal mammary gland growth, development and sexual maturation. Accumulated studies reported that autocrine hGH is an orthotopically expressed oncogene and could induce normal mammary epithelial cell malignant transformation. Previously, our group reported autocrine hGH promotes mammary epithelial cell epithelial‐mesenchymal transition (EMT), facilitating invasion and increasing MMP activity. However, how hGH regulates EMT and invasion remains largely obscure. miRNAs are widely reported to involve in multiple processes of cancer. To determine whether autocrine hGH promotes EMT through miRNAs, we performed microarray profiling using MCF‐7 cells stably expressing normal or translation deficient hGH. We found that miR‐96‐182‐183 cluster exhibited highest response to autocrine hGH stimulation. We further observed that forced expression of miR‐96‐182‐183 conferred epithelial MCF‐7 cells with mesenchymal phenotype and invasive behaviors both in vitro and in vivo. Moreover, we revealed that miR‐96‐182‐183 promotes EMT and invasion by directly and simultaneously suppressing Breast Cancer Metastasis Suppressor 1‐like (BRMS1L) gene. In addition, miR‐96 and miR‐182 target GHR, introducing a potential negative feedback loop in the hGH‐GHR signaling pathway. Mechanically, we found that autocrine hGH stimulates miR‐96‐182‐183 expression and facilitates EMT and invasion via STAT3 and STAT5 signaling pathway. Clinically, we inspected that miR‐96‐182‐183 expression is remarkably enhanced in metastatic breast tumors. Together, we elucidate one novel hGH‐GHR‐STAT3/STAT5‐miR‐96‐182‐183 cluster‐BRMS1L‐ZEB1/E47‐EMT‐invasion axis, which sheds new insights into understanding the mechanism of autocrine hGH stimulated EMT and invasion of breast cancer cells.