bluuebunny/crossref_metadata_2025 huggingface dataset:
Ontogenic analysis of the PACAP receptor, PAC1R, in mouse major salivary glands (916.1)
Authors: Nonaka Naoko , Nakamura Masanori
Published: June 2021

The control of saliva secretion is mainly under the autonomic nervous control. Pituitary adenylate cyclase‐activating polypeptide (PACAP) is now recognized as the multi‐functional neuropeptide in various organs. Previous our study indicated the localization of PAC1R in adult mouse major salivary glands. In this study, we examined the ontogenic expression of PAC1R in mouse salivary glands. Salivary glands at 0, 3, 5, 7, 14 and 21 days postnatal male C57BL/6 mice were used for the immunohistochemical detection of PAC1R. In parotid gland, the acinus was not clearly formed and PAC1R was not detected until 14 days after birth. At 14 days, PAC1R was detected in the striated duct. In sublingual gland, Acinus and duct were formed at 0 days after birth. PAC1R was constantly expressed in the striated duct from 0 day postnatal stage. In the case of submandibular gland, granular duct was not clearly identified and PAC1R was localized in striated first detected in striated duct until 21 days after birth. After 21 days, PAC1R was clearly detected at pillar cells in granular duct. Some of cells in the duct showed intense immunoreactions of PAC1R. These results indicated that PAC1R was expressed in the striated duct of parotid and sublingual glands and that the localization of PAC1R was changed with the formation of granular duct. The precise study might be necessary to clarify the function of pillar cells by examining the shift of PAC1R‐positive cells in submandubular gland.
Gluten‐like cellular response of maize prolamins in intestinal biopsies and Caco‐2 cells (916.6)
Authors: Ortiz‐Sánchez Juan , Calderón de la Barca Ana , Mata‐Haro Verónica , Hernández‐López Jesús , Cabrera‐Chávez Francisco
Published: June 2021

Celiac disease (CD) is an autoimmune‐mediated enteropathy triggered by wheat gliadins in genetically prone individuals. Some celiac patients are affected by maize zeins, possibly by a gliadin‐like immune response. The immunogenic peptides of gliadin (G33mer) and zein (Z34mer) were used to evaluate activation of cultured intestinal biopsies of patients with different gastrointestinal diseases. Additionally, changes in the permeability of a monolayer of Caco‐2 cells challenged with peptides of digested gliadin (GdDig) and zeins (ZeDig), were evaluated. Pathological report indicated inflamed duodenal mucosa (7), mucosal atrophy (2) and normal mucosa (1) in 10 patient biopsies. Interferon‐gamma (IFN‐γ) and interleucin (IL‐2) specifically increased in atrophic mucosa by effect of G33mer, while the Z34mer induce a smaller increase in cases of atrophic and inflamed mucosa compared to control. ZeDig peptides and G33mer induced a higher permeability of the monolayer of Caco‐2 cells, as compared to that induced by GdDig or Z34mer. Zeins could have an effect on disrupted mucosa similar to gliadins but not necessarily by induction of the initial damage.
The cervical cutaneous and subcutaneous structures in songbirds and their possible roles in vocalization (918.10)
Authors: Cozic Amanda , Suthers Roderick , Homberger Dominique
Published: June 2021

Avian vocalizations are the product of the sound‐generating syrinx and the sound‐resonating suprasyringeal structures and organs of the neck. Whereas the sound‐resonating role of the oro‐pharyngo‐esophageal cavity has been analyzed, this is not the case for the rest of the cervical structures. We therefore undertook a functional‐morphological analysis of the soft‐tissue organs of the neck in two songbird species, the Northern Cardinal, Cardinalis cardinalis, and the House Sparrow, Passer domesticus, by using micro‐dissection to create the soft tissue parameters to be applied to a virtual 3D model that was generated from a CT scan. The soft‐tissue organs of the neck are arranged in concentric layers: (1) The skin with dorsal and ventral feather tracts (pterylae) separated by large featherless lateral apteria; (2) the constrictor fascia and muscle; (3) the longitudinal fascia with longitudinal subcutaneous muscles; and (4) the trachea and esophagus. This system creates a cervical coelom that allows the esophagus, crop, and trachea some mobility relative to the cervical vertebral column. In addition, the integument and fascia may serve as a drum skin over a resonating chamber created by the oro‐pharyngo‐esophageal cavity. The tension of the skin and fascias can be modified by the varying curvature of the vertebral column during singing, which may tune the resonating chamber like the tunable Asian Indian drums (tablas and mridangams).

Grant Funding Source: Supported by NIH grant NINDS R01 NS029467 to RAS and the LSU Foundation account “Functional Morpholo
Energetic costs of feeding behaviors in the aye‐aye, Daubentonia madagascariensis (918.14)
Authors: Toler Maxx , Hanna Jandy , O'Neill Matthew , Wall Christine
Published: June 2021

The aye‐aye is a 2.5‐3kg lemur that has a specialized feeding apparatus featuring rodent‐like incisors which they use during extractive foraging on wood‐boring larvae and seeds. To evaluate these behaviors, we measured the energetic costs of feeding on whole seeds and wood gnawing in aye‐ayes (n=4) using open‐flow respirometry. Rates of oxygen consumption were measured during stable periods of rest and feeding for each experiment. The net feeding costs reported are the cost of feeding minus the energy expended during rest in Watts. The mean net feeding rate was 14.7W (SD 3.7) during wood gnawing and 5.6W (SD 2.5) during seed eating. Our results show that wood gnawing was significantly more costly (Wilcoxon, p<0.0001) than seed predation for aye‐ayes. These differences are partially caused by variation between material properties of food objects and wood, but may also be mediated by differences in feeding behavior; wood gnawing aye‐ayes likely recruit postcranial muscles to maintain tooth contact with a large, immobile surface while closing their jaws. While these data suggest that extracting larvae is a costly behavior, insect larvae are rich in protein and fat and are less seasonal than the seeds consumed by wild aye‐ayes. Specialization of the incisor teeth to maximize bite pressure and resist wear are likely adaptations to high cost, high reward foraging behaviors in the aye‐aye.
Comparisons of mid‐facial contour shape throughout early development (918.24)
Authors: Wheat Amber
Published: June 2021

Craniofacial shape is often used as an indicator of ancestry within the field of biological anthropology. It is well known that adults portray geographical variation in the shape and relative size of dimensions in the face, but little is known about the degree to which biological and environmental factors affect development in craniofacial shape. That is, are the traits that researchers use to identify human groups evident early in life, and what factors might influence these dimensions? One method for ascertaining the developmental patterns that produce craniofacial variation is to assess the morphological integration of the mid‐face. While previous research has investigated the integration of the mid‐face region in adults, the patterns of mid‐face integration from an ontogenetic perspective is not well understood. This study investigates the ontogenetic patterns of mid‐facial contour in two human populations to better understand how development contributes to population divergence in mid‐face shape. One hundred three‐dimensional computed tomographic (CT) images of the heads of juveniles (ages 5‐16) from two different populations were utilized: American “whites” and “blacks”. Each population was divided into three different age groups. 3D coordinate locations of landmarks defining mid‐face contour were recorded. Euclidean distance matrix analysis was used to analyze differences in mid‐face shape between the two human groups across all three ontogenetic groupings. Preliminary results indicate significant differences in mid‐face contour between population groups, suggesting that development contributes to population divergence.
Higher lumbar lordosis among women: a study examining lumbar angle and dorsoventral wedging of vertebral bodies and discs in standing and supine radiographs (919.16)
Authors: Bailey Jeannie , Been Ella , Kramer Patricia
Published: June 2021

Studies have suggested that the lumbar spine of humans is sexually dimorphic with women exhibiting higher dorsoventral vertebral wedging and more lordotic curvature than men. Greater lumbar curvature is proposed to allow women to center their mass while bearing a fetal load during pregnancy. These studies are based, however, on measurements from osteological collections that cannot account for disc wedging or the effect of standing. We collected 138 radiographs of males (n=66; supine, n=20; standing, n=46) and females (n=70; supine, n=36; standing, n=34), ranging in age from 8 to 56 years. We measured lumbar angle between the cranial endplates of L1 and S1, as well as wedging angles of each vertebra and disc. Results showed lumbar angle was significantly greater in females than males (f, 61.301°, sd 2.34°; m, 53.32°, sd 1.88°; p=0.002), after adjusting for position and age. Also, that all of the vertebral bodies were significantly more dorsoventrally wedged in females and that none of the discs were significantly different between sex (mean difference between females and males: 1.28° for L1, 1.56° for L2, 1.30° for L3, 1.83° for L4, and 2.73° for L5). We concluded that while standing, women do have greater lumbar lordosis than men and that each lumbar vertebra is more dorsoventrally wedged.
Role of hepatic cytochrome P450 oxidoreductase in bone development (921.1)
Authors: Kellogg Dean , Tang Kejian , Roman Linda , Fajardo Roberto , Masters Bettie Sue , Panda Satya
Published: June 2021

Cytochrome P450 oxidoreductase (POR) is the only known electron donor for Cytochromes P450 (CYPs), heme oxygenase (HO), and dehydrocholesterol reductase in the endoplasmic reticulum (ER). Hepatic CYPs are essential for the metabolism of cholesterol, steroids, sex hormones, and xenobiotics, such as drugs and toxins. Thus, alterations in POR activity will have multiple effects on the physiological activity of multitudinous metabolites. Human mutations in POR have been reported and present as altered steroidogenic profiles, severe craniofacial and bone defects resembling Antley‐Bixler syndrome (ABS). Liver‐specific POR null (LCN) mice were examined to study endo‐ and xeno‐biotic metabolism. Surprisingly, previous studies demonstrated only minor changes in plasma and intestinal lipid content of LCN mice, suggesting that lipid homeostasis is maintained despite significant hepatic steatosis. Here, we show for the first time that the deletion of POR from the liver leads to decreased bone volume in both cortical and trabecular compartments, as measured by micro computed tomography analysis, but no notable skull defects. In bone‐specific POR null mice, however, we have previously observed defects in skull morphology and trabecular bone density. This difference suggests that the action of hepatic CYP‐derived circulating metabolites is critical for normal bone development and is distinct from bone‐ specific CYP activity. There are several potentially important metabolites that could affect bone homeostasis and require hepatic CYP hydroxylation such as vitamin D, retinoic acid and steroid hormones. These data highlight the importance of both systemic and local POR activity in healthy bone development.

Grant Funding Source: Supported by NIH grant GM081568 to BSSM and SPP
A one month old infant misdiagnosed with parietal bone fracture and child abuse (922.3)
Authors: Collins James , Saxton Ernestina , Carnes Alfred
Published: June 2021

INTRODUCTION. A month‐old female infant was brought to the emergency room because she would not take in fluids. Her mother indicated the infant had a birth history of a traumatic brain injury stemming from a difficult instrumental delivery. A nurse assigned to care for the infant observed what appeared to be blood on the right ear. She immediately alerted the pediatrician on duty of a possible case of child abuse. The pediatrician concurred and ordered an anterior‐posterior chest radiograph and anterior posterior and lateral skull radiographs. RESULTS. The chest radiograph was interpreted by the hospital radiologist as being positive for fractures of the ribs and the skull radiographs as being positive for bilateral parietal bone fractures without soft tissue swelling. Three‐dimensional computerized axial tomography was then performed and interpreted as positive for bilateral parietal bone fractures. Although the infant was not in acute distress, she was hospitalized with the suspected diagnosis of child abuse. An outside radiologist was consulted finding no evidence of fractures and/or swelling over the sites dictated by the hospital radiologist. The outside radiologist found an anomalous parietal bone suture misinterpreted as the parietal bone fracture. CONCLUSIONS. The purpose of this presentation is not to review the division of the parietal bone but to inform physicians and health professionals that anomalous parietal bone sutures are rare and often misdiagnosed as fractures.
Analysis of yeast SUMO ligase MMS21‐dependent genome maintenance pathway (925.4)
Authors: Liang Jason , Lee Nancy , Albuquerque Claudio , Putnam Christopher , Kolodner Richard , Zhou Huilin
Published: June 2021

Genome instability often characterizes many cancers and is indicative of high frequency of mutations and genome rearrangements. Several pathways, including the DNA damage checkpoint, play a role in maintaining the stability of the genome by suppressing gross chromosomal rearrangements (GCRs). Additionally, eukaryotic genomes contain numerous repetitive sequences, segmental duplications and regions of copy number variation, which influence the outcomes of gross chromosomal rearrangements.

Using a genetic assay in Saccharomyces cerevisiae to screen for suppressors of GCRs, we recently reported that Mms21‐dependent sumoylation specifically suppress GCRs mediated by homeologous sequences. These types of duplication‐mediated GCRs are attributed to Rad52‐dependent non‐allelic homologous recombination (NAHR). We will present findings that show unexpectedly, the DNA damage checkpoint to be required for the formation of duplication‐mediated GCRs in the absence of Mms21‐dependent sumoylation. The canonical DNA damage checkpoint genes MEC1, RAD24, MEC3, RAD9 and RAD53 are required for formation of duplication‐mediated GCRs in the mms21 mutant, while TEL1, MRC1 and CHK1 are minimally involved. Among the checkpoint genes, mutation of RAD9 strongly suppresses GCRs formed in the mms21 mutant and this suppression is independent of either SIZ1 or SIZ2, which encode two other SUMO ligases in yeast. We will also present findings on the role of DNA damage checkpoint in regulating the SUMO proteome. These results show that the outcomes of GCRs are dynamic and are governed by a fine balance of distinct genome maintenance pathways.

Grant Funding Source: GM080469, T32CA009523 ,LICR
Determining the genetic basis of equine deafness via candidate gene studies (928.3)
Authors: Innamorati Katherine , Lipsett Koren
Published: June 2021

White fur or coat pigmentation and the presence of blue eyes have been linked to cochleosaccular deafness in multiple mammalian species. At this time, no hereditary explanation has been presented for this phenotype as it appears in horses. A candidate gene study was established to determine the genetic basis for this disorder in a pedigree of Spanish Mustangs. KIT, a receptor tyrosine kinase implicated in multiple mammalian deafness studies, was sequenced in both the affected Spanish Mustangs and related members of the pedigree. A mutation (KI16 + 1037A) in the 5’ splice junction of exon 17 known to cause the sabino coat pattern appeared to be unlinked from the deafness phenotype. Sabino coloration is distinguished as a white coat, ranging from minimal white splashes to an entirely white mammal. Future research will focus on the sequencing of additional candidate genes affecting melanocyte migration and melanin production, including EDNRB, MITF, PMEL17, and SOX10.

Grant Funding Source: Supported by HHMI
Minor groove geometry determines DNA exclusion from nucleosomes (929.1)
Authors: Grilley Dan
Published: June 2021

Like all eukaryotes, the meters of DNA in each of our cells must be compacted many thousands of times to fit inside the nucleus. The first level of compaction, spooling of DNA onto a nucleosome, involves the tight wrapping of DNA around eight histone proteins. Sharply bending DNA to wrap it into nucleosomes comes at an energetic cost related to the stiffness of the DNA. Nucleosomes prefer to form on DNA sequences that have a 10 bp periodicity between AA/AT/TA steps, with CC/CG/GC steps exactly out of phase. The sites of the AA/AT/TA and CC/CG/GC steps exactly correspond to locations of the highest DNA kinking, suggesting that these sequences represent anisotropically bendable sequences. Yet, inclusion of multiple AA steps, in the form of a poly(dA:dT) elements, results in exclusion of the DNA from nucleosomes in vivo. We seek to understand what unique chemical features of these poly‐dA elements are necessary for their exclusion from nucleosomes. Poly‐dA elements are known to adopt a unique structure; characterized crystallographically as a narrowed minor groove and by CD as a double peaked spectrum. Using CD spectroscopy and non‐natural nucleotides, we show that correct minor groove geometry at the base pair level is necessary for formation of the unique poly‐dA structure. Using in vitro competitive nucleosome reconstitution assays, we have further shown that non‐natural DNA sequences that adopt the unique poly‐dA structure are excluded from nucleosomes. Since poly‐dA like sequences are excluded from nucleosomes, this suggests that the unique structure formed by these sequences is energetically stiffer than generic sequence DNA.
The role of the PAR2‐beta‐arrestin signaling axis in Alternaria and cockroach‐induced allergic asthma (934.3)
Authors: Yee Michael , Nichols Heddie , Pal Kasturi , Sy Sireena , Defea Kathryn
Published: June 2021

A common cause of asthma is the exposure to allergens such as the household fungus, Alternaria alternata (AltA), and the feces of the common cockroach, Blattella germanica (BG), which contain proteases that activate Protease‐activated‐receptor‐2 (PAR2). In several murine models of asthma, PAR2 activation has been shown to be pro‐ and anti‐inflammatory. Recently, our lab has shown that the pro‐inflammatory pathway is mediated by beta‐arrestins, which act as both terminators of GPCR signaling through G‐proteins, and as scaffolding proteins that signal independently of G‐proteins. In contrast, the anti‐inflammatory pathway requires G‐protein signaling. We hypothesize that proteases from AltA and BG extracts will promote activation of the beta‐arrestin‐2 dependent inflammatory pathway in murine models that we have previously observed with intranasal administration of Par2 peptidomimetic agonists. Here, we have performed flow cytometric analyses of immune cell populations and analyzed histological samples in physiological mouse models of AltA and BG‐induced allergic asthma. Our results show that airway inflammation induced by AltA and BG are abolished in beta‐arresitn‐2‐/‐ and PAR2‐/‐ mice. Furthermore, AltA and BG proteases activate downstream signaling mechanisms in a beta‐arrestin‐2‐dependent way. This suggests that the proteases can activate the PAR2‐beta‐arrestin signaling axis, which in turn regulates immunological responses in the airway.
Transcriptional changes in gamma‐secretase conditional knockout mice (934.5)
Authors: Arbella Jennifer , Yu Gang , Dries Daniel
Published: June 2021

gamma‐secretase is an intramembrane protease composed of four transmembrane proteins: presenilin, nicastrin, Pen‐2, and Aph‐1. Several studies have examined the role of gamma‐secretase in neurons but none have done so in non‐neuronal cells. Therefore, our lab developed a conditional knockout mouse in which gamma‐secretase was conditionally deleted from the myelinating cells of the brain. Here, we use quantitative PCR to confirm the specificity of the conditional deletion and to probe a series of putative genes for their altered expression in conditional knockout mice. Moreover, we begin to look at transcriptional changes in the conditional knockout animals using using RNA‐seq.

Grant Funding Source: Supported by National Institute of Neurological Disorders and Stroke grant 1R01NS079796
Live cell monitoring of gene modulation provides a better understanding of gene function (946.12)
Authors: Weldon Don , Williams Yuko , Koong Victor , Huryn‐Selvar Karyn
Published: June 2021

One of the most common methods for understanding gene function within a live cell is modulating the gene through knockdown or over‐expression systems. These experiments provide information regarding the role a particular gene plays within a given biological process. Unfortunately, determining the modulation in gene expression is currently performed at the population level as in the case of qPCR and or is ultimately an endpoint determination as in the case of RNA FISH.

We propose a new method for determining the RNA expression levels of target genes using a nanoparticle technology which can be used in live cells without the need for transfection reagents. Live cell gene expression detection affords the ability to monitor changes in expression levels over time. More data can be collected over the course of a single experiment at the cellular level while monitoring gene expression in a dynamic manner as opposed to studying many samples at varied time points in the case of RT‐PCR or FISH.

Here we present data illustrating the benefits of detecting gene expression in live cells while modulating both Survivin and ER alpha genes. U2OS cells which have an ER alpha knock‐in under the control of an inducible promoter were monitored using the nanoparticle detection method prior to and during the induction process enabling the monitoring of the gene modulation. Knockdown experiments were also performed using Survivin as a target gene to illustrate down regulation of genes and the ability to monitor the loss of function as well.

Monitoring the modulation of genes within live cells can assist in determination of gene function by allowing for the collection of many more data points within a single experiment without the need for duplicate samples at various time points. Furthermore, as this method is non toxic the live cells can be further characterized or used in subsequent experiments providing even more data from the same cells.
Quantifying the two populations of dual oriented P6 in nontypable Haemophilus influenzaeand Pal in Escherichia coli (948.6)
Authors: Shaw Juliana , Schmidt Rachel , MacPherson Victoria , Pichichero Michael , Michel Lea
Published: June 2021

Nontypable Haemophilus influenzae (NTHi) causes otitis media and other respiratory diseases in people of all ages, conferring the need for a vaccine against this bacterium. P6 is a lipoprotein and a vaccine candidate for protection against NTHi. Recent studies show that P6 is inserted into the outer membrane of NTHi in two distinct orientations‐ facing out toward the extracellular space and in toward the periplasm (Michel et al., J of Bacteriology 195: 3252). To further investigate the two orientations of P6, we utilized a cell‐impermeable biotinylating reagent (NHS‐LC‐LC‐biotin) and streptavidin‐agarose beads to separate the outward and inward facing P6. We then used standard protein detection methods to quantify the two populations. Results showed that the great majority of P6 faces in toward the periplasmic space of NTHi. Similar methods were used to quantify the two populations of Pal, P6’s homologue in Escherichia coli. Our results showed that, like P6, Pal is mostly oriented facing in toward the periplasmic space of the bacterial cell. We propose that, in light of our results, there may be more dual oriented lipoproteins in biology.

Grant Funding Source: This study was supported by NIH NIDCD RO1 08671 (to MEP) and the Rochester Institute of Technology.
Prevention of vascular disease of vegetable oils in HUVEC (959.15)
Authors: Kim Kyoung Kon , Kim Dae Jung , Kang Yun Hwan , Lee Sung Mee , Choe Myeon
Published: June 2021

To confirmation of anti‐atherosclerosis effect by treatment of various vegetable oils, we determined the activity of cell proliferation, cell migration and NO production in HUVECs. The results of proliferation assay on vegetable oil treated HUVEC showed that, TNF‐α increased the cell proliferation and the pine nuts 0.1 μl/ml, safflower seeds 0.1 μl/ml, ginseng seeds 0.1 μl/ml inhibited the cell proliferation activity by 89.4%, 90.34%, and 91.4%, respectively, but, other vegetable oil did not affected. In cell migration assay, classified the cultured cell plates with the tip scratches and then treated with TNF‐α to stimulate cell migration. Red pepper seed, green tea seeds, citron seeds, and ginseng seeds oils of 12 vegetable oils inhibited TNF‐α‐stimulated cell migration. Nitric oxide synthase activation effect of vegetable oils was measured by NO production assay. Ginseng seeds 0.1 μl/ml, safflower seeds 0.1 μl/ml, chinese pepper 0.1 μl/ml increased the NO production by 131.43%, 129.63%, and 129.52%, respectively, but other oils did not affected. Therefore, our results suggest that further studies are required to verify the prevention of vascular disease by vegetable oil.
Anti‐diabetes effects of Smilax china L. extract (959.17)
Authors: Kang Yun Hwan , Kim Dae Jung , Kim Tae Woo , Kim Kyoung Kon , Choe Myeon
Published: June 2021

Smilax china L., a native plant found in Asian countries, has several medicinal properties including antioxidant, anti‐inflammatory, and anti‐cancer effects. Although the root of the plant is commonly used as traditional herbal medicine in Korea and China, the medicinal properties of the leaves have not gained the same attention. In this study, we analyzed the antioxidant activity, α‐glucosidase inhibitory effect and lipid accumulation inhibition effect of Smilax china L. leaf water extract (SCLE) and its solvent fractions. SCLE was fractionated by using a series of organic solvents, including ethylacetate (EA) and n‐butanol (BuOH). The EA fraction had the highest total polyphenol content (440.20 ± 12.67 mg GAE/g) and total flavonoid content (215.14 ± 24.83 mg QE/g). The radical scavenging activity IC50 values of the EA fraction for 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) and 2,2‐azino‐bis‐(3‐ethylbenzthiazoline)‐6‐sulfonic acid (ABTS) were 0.022 mg/mL and 0.13 mg/mL, respectively. Further, SOD‐like activity and reducing power values of the EA fraction were higher than those of the other fractions. However, both the α‐glucosidase and lipid accumulation inhibition assays showed that the BuOH fraction (83.35 ± 4.18% at 1 mg/mL) and water extract (11.27 ± 2.67%) were more effective than the EA fraction (64.13 ± 6.35%, and 45.66 ± 7.20%). These results provide new insights into the potential anti‐diabetic and anti‐obesity effects of Smilax china L. leaf.
Inhibitory effects of pentagalloyl glucose from winged sumac (Rhus copallinum) on albumin glycation and the formation of advanced glycation endproducts (975.4)
Authors: Ma Hang , Liu Weixi , Navindra Seeram , Dain Joel
Published: June 2021

Glycation is a spontaneous process typically initiated with a condensation reaction between a reducing sugar and a protein, leading to the formation of Advanced Glycation Endproducts (AGEs). The in vivo accumulation of AGEs is associated with many of the chronic conditions associated with diabetes including atherosclerosis, renal failure, and cataract formation. The objective of this study was to evaluate the inhibitory effects of pentagalloyl glucose (PGG), a polyphenol isolated from a native North American medicinal plant, winged sumac (Rhus copallinum), on individual stages of protein glycation using a combination of analytical methods including fluorescence spectroscopy, HPLC, circular dichroism and MALDI‐TOF. The inhibitory effects of PGG on both early and middle stages of glycation were evaluated using bovine serum albumin and D‐fructose serving as model protein and glycating agent, respectively. PGG inhibited the formation of AGEs and was 50‐fold more potent compared to the positive control, aminoguanidine (IC50 = 5.3 vs. 250 µM). Moreover, PGG was effective in the prevention of protein cross‐linking formation shown by G.K. peptide‐ribose assay, indicating that PGG not only inhibited the early and middle stages of the Maillard reaction but also the late stage of glycation. The inhibitory effects of PGG on glycation warrants further investigation as this compound may have nutraceutical applications for the reduction of glycation products in patients with diabetes and other conditions that promote hyperglycemia.
Exploitation of host cell processes for bacterial cell‐to‐cell spread (99.4)
Authors: Theriot Julie , Rengarajan Michelle
Published: June 2021

The ubiquitous food‐borne bacterium Listeria monocytogenes causes spontaneous abortion in pregnant women and meningitis in immune‐compromised hosts. Bacteria in the intestine directly invade intestinal epithelial cells, but systemic dissemination requires that the bacteria use actin‐based motility to move into macrophages that carry them through the bloodstream to distal sites, and then that they cross the endothelial barrier to invade protected tissues such as the placenta and meninges. In tissue culture experiments, we have used fluorescence microscopy and flow cytometry to confirm that L. monocytogenes can directly invade and spread through a monolayer of human umbilical vein endothelial cells (HUVEC). We performed a targeted siRNA screen to identify host cell factors that modulate bacterial invasion and spread. Knocking down focal adhesion components including integrin, paxillin, and vinculin specifically increased bacterial invasion, suggesting that focal adhesions may normally inhibit invasion. We have also demonstrated that L. monocytogenes in infected macrophages can be robustly transferred to uninfected HUVEC via direct heterotypic cell‐to‐cell spread. Using siRNA screening, we have identified a number of cytoskeletal factors, including Arp2, capping proteins, Rac, Diaph2, and ZO1, which, in recipient cells, appear to decrease heterotypic bacterial transfer, suggesting that the endothelial cortex and tight junctions between endothelial cells may influence macrophage‐to‐endothelial transfer of bacteria.
Overexpression of the novel gene CIA7 in Chlamydomonas reinhardtii (995.3)
Authors: Pedraza Ricardo , Ynalvez Ruby
Published: June 2021

Heavy metal pollution has become a significant environmental and public health problem that the world faces. C. reinhardtii is an excellent model organism that has often been used in studies of heavy metal tolerance. This alga contains the novel gene CIA7 that possibly encodes a protein that may play a role in metal binding. This gene was isolated from inorganic carbon acquisition mutants. By cloning and overexpressing CIA7 in C. reinhardtii, it can be determined if CIA7 is critical in C. reinhardtii’s metal toxicity tolerance. This gene of interest was first extracted from the alga and then amplified using PCR. Vectors pENTR™/D‐TOPO® (Invitrogen) and psL72 plasmids were used for cloning CIA7 in E.coli. C.reinhardtii cells were then transformed through electroporation. Growth analysis between wild type C.reinhardtii D66 and transformants at different metal concentrations were done to determine the effect of CIA7 on C.reinhardtii under metal stress. If a link between this protein and heavy metal tolerance is identified, this could potentially lay the foundation for its use in heavy metal pollution.

Grant Funding Source: Supported by: Norman Hackerman Advanced Research Program
Role of lysophosphatidic acid in traumatic brain injury: anti‐LPA antibodies are neuroprotective after experimental TBI (999.3)
Authors: Wojciak Jonathan , Sabbadini Roger , Crack Peter , Morganti‐Kossmann Cristina , Zhang Moses , Pebay Alice , Conquest Alison , Morris Andrew
Published: June 2021

Lysophosphatidic acid (LPA) is a bioactive phospholipid with a potentially causative role in neurotrauma. LPA receptors are increased after human brain injury and LPA levels are upregulated in the CSF and blood of traumatic brain injury (TBI) patients. Further, LPA levels correlated with initial injury severity using GCS, GOSE and ISS clinical assessments. Blocking LPA signaling with the LPA‐directed monoclonal antibody, B3/Lpathomab, was neuroprotective in a mouse controlled cortical impact model of TBI. B3 significantly reduced lesion volume measured both by standard histological assessments and MRI. Diminished tissue damage coincided with improvement in functional outcomes. B3 was anti‐inflammatory by limiting cerebral mRNA expression of pro‐inflammatory cytokines such as interleukin‐1b. These data strongly suggest that LPA dysregulation/upregulation is causal to injury progression following TBI, and that interventions to reduce LPA may have a significant impact in mitigating early and late phases of neurotrauma. We propose that anti‐LPA mAbs could satisfy a critical unmet need by limiting the initial neuronal damage and inflammatory processes while subsequently stimulating regenerative processes to optimize long‐term functional outcomes after neurotrauma. Importantly, LPA may also be a novel biomarker for TBI.

Grant Funding Source: Lpath, Inc.
The Mechanism for Tau Tangles (LB102)
Authors: Ambessie Sallem , Shire Sequoia , Simpson Nick , Yenew Mihret , Dix Randy , Slawson Chad
Published: June 2021

Alzheimer’s disease (AD) affects more than 5.2 million people in the US. The disease is characterized by the presence of “Tau Tangles”, which is the aggregation of the protein Tau. The Tau protein is expressed in the nervous system and helps stabilize microtubules, which help establish cell shape and serve as tracks for neuronal transport. The Tau protein is regulated through phosphorylation and in patients with AD, the Tau protein is hyperphosphorylated, which can lead to the formation of “Tau Tangles”. These tangles are made up of free floating Tau and can interfere in the neuron communication because of a structural change in the protein due to the phosphorylation. In a normal mature neuron, tubulin is present in over tenfold excess of the Tau protein, and almost all Tau protein is bound to microtubules. In AD, the ratio between free floating Tau and microtubule bound Tau increases. In neurons affected by AD, hyperphosphorylated and glycosylated Tau disengages from microtubules and disrupts microtubule organization. Tau kinases phosphorylate Tau; however the regulation of Tau phosphorylation is still unknown. The Olathe North SMART Team (Students Modeling A Research Topic) modeled the Tau kinase using 3D printing technology.

Grant Funding Source: Supported by grants from the NIH‐SEPA and NIH‐CTSA.
Getting “Rit‐R” Iron in the Cell: The Role of RitR in Reducing Iron Transport into Streptococcus pneumoniae. (LB111)
Authors: Allbee B. , Anderson N. , Blank D. , Bolgert J. , Bothe A. , Britt N. , Geiser E. , Hammer T. , Janecek E. , Kalmer I. , Lawniczak J. , Lesiecki M. , Levy J. , Marshall M. , Meaux N. , Ruzicka M. , Temmer J. , Tiffany K. , Butt A. , Dyke S. , Vandenberg B. , Wankowski J. , Zeitlow E. , Tiffany Karen , Silvaggi Nicholas
Published: June 2021

According to the World Health Organization, pneumonia is the leading cause of death in children worldwide, and infection of lung tissue by Streptococcus pneumonia causes the bulk of bacterial pneumonia cases in children. The atypical response regulator, RitR (Repressor of iron transport Regulator), helps S. pneumoniae survive in the hostile oxidizing conditions in the lungs. RitR has two domains, a DNA‐binding domain (DBD) and an aspartate‐less receiver domain (REC). In its “inactive” form, these domains are docked (i.e., close together) and the DBD is unable to bind DNA. In its “active” form, the domains are undocked; the DBD is freed from the REC. The “active” form dimerizes and can bind to DNA to turn off the iron transport genes. To study the changes that occur when RitR is activated, the Cedarburg High School SMART (Students Modeling A Research Topic) Team used 3D printing technology to model inactive RitR and a hypothetical active RitR dimer. RitR helps the bacterium survive in the oxygen‐rich environment in lungs by stopping iron transport into the bacterial cell. If iron is transported into the cell, oxygen forms reactive oxygen species that damage and kill cells. S. pneumonia cells without RitR are unable to infect lung tissue, so RitR is a potential target for drug design.

Grant Funding Source: Supported by a grant from NIH‐CTSA.
Somatic mutant alleles of POLE found in human cancers suppress proofreading and replication fidelity in vitro (LB121)
Authors: Henninger Erin , LeCompte Kim , McBride Christine , Bunnell Bruce , Pursell Zachary
Published: June 2021

While it is now appreciated that tumor cells contain thousands of mutations, identifying their source(s) has remained problematic. Whole genome sequencing from colorectal and endometrial cancers recently identified mutations in DNA polymerase (Pol) epsilon, one of three main eukaryotic nuclear replication Pols. These mutations largely reside in the exonuclease domain, are associated with a hypermutated phenotype, and are predicted to promote tumorigenesis. However, the effects of these somatic mutations on enzyme function have not yet been studied. To address this, we introduced several of these mutations in recombinant human Pol ε and measured their effects on replication fidelity in vitro. While the mutants retained wild type DNA synthesis activity, 3'‐5' exonuclease proofreading was suppressed. Surprisingly, this suppression was variable, ranging from ~2‐fold to almost complete ablation, and resulted in increases in frameshifts and base pair substitutions in an in vitro forward mutation assay. Mutants also had increased ability to bypass abasic sites. Exonuclease inactivation of one POLE allele in human cells resulted in decreased sensitivity to H2O2 treatment. These results suggest that Pol ε mutant alleles may promote viability and mutagenesis, and possibly tumorigenesis, in part through bypass of oxidative stress‐induced DNA lesions.

Grant Funding Source: NIH RR020152
The Effect of Artesunate on Myeloperoxidase Activity of Human Polymorphonuclear Neutrophil. (LB134)
Authors: MINARI JOSEPH
Published: June 2021

Myeloperoxidase is the most abundant enzyme found in the polymorphonuclear neutrophil and is known to play a central role in the host defense system of the leukocyte. The enzyme has been reported to interact with some drugs to generate free radical which inhibits its activity. This study investigated the effects of artesunate on the activity of the enzyme and the subsequent effect on the host immune system. In investigating the effects of the drugs on myeloperoxidase, the influence of concentration, pH, partition ratio estimation and kinetics of inhibition were studied.

This study showed that artesunate is concentration‐dependent inhibitor of myeloperoxidase with an IC50 of 0.078mM. Partition ratio estimation showed that 60 enzymatic turnover cycles are required for complete inhibition of myeloperoxidase in the presence of artesunate. The influence of pH on the effect of artesunate on the enzyme showed least activity of myeloperoxidase at physiological pH. The kinetic inhibition studies showed that artesunate caused a competitive inhibition with an increase in the Km value from 0.12mM to 0.26mM and no effect on the Vmax value. The Ki value was estimated to be 2.5mM. The results obtained from this study show that artesunate is a potent inhibitor of myeloperoxidase and it is capable of inactivating the enzyme.

It is considered that the inhibition of myeloperoxidase in the presence of artesunate as revealed in this study may partly explain the impairment of polymorphonuclear neutrophil and consequent reduction of the strength of the host defense system against secondary infections.
Long intrahemispheric pathways of the human cerebral cortex visualized by dissection (LB21)
Authors: Turner Blair , Knapp Margaret
Published: June 2021

Long intrahemispheric pathways of the human cerebral cortex visualized by dissection In order to visualize in the human brain analogous corticocortical pathways linking the parietal and frontal lobes that have been found experimentally in the monkey, we dissected fixed (donated) human hemispheres. Our findings showed four such pathways: Two medial to the corona radiata, and two lateral to the corona radiata. Medial: 1. The fimbria link the hippocampus to the mammillary body. 2. The cingulum bundle connects the medial occipital lobe to the subcallosal cingulate gyrus. Lateral: 3. The uncinate fasciculus links the posterior superior temporal gyrus with the inferior prefrontal cortex. 4. The inferior longitudinal fasciculus connects the anteromedial occipital lobe to the inferior temporal gyrus. We conclude that the posterior cerebral cortex projects heavily to frontal cortex by discrete axonal pathways that connect sensory information to motor mechanisms.
β‐like Importins mediate the nuclear translocation of JNK1/2 and p38α/β (LB219)
Authors: Zehorai Eldar , Seger Rony
Published: June 2021

The rapid nuclear translocation of signaling proteins upon stimulation plays an important role in the regulation of de‐novo gene expression. We have studied the stimulated nuclear shuttling of JNK and p38 MAPKs, and found that they translocate into the nucleus in NLS‐ or NTS‐independent manners, unrelated to their catalytic activity. In addition, we found that this translocation is mediated by three β‐like importins (Imp 3, 7 and 9). Knocking down these importins inhibits the stimulated nuclear translocation of the MAPKs, and thereby, activation of their transcription factors targets. We demonstrated that this process involves the binding of either JNK1/2 or p38α/β to Imp7 or Imp9 and requires their further association with Imp3. These heterotrimers then translocate to the nuclear envelope, where Imp3 remains, while Imp7 or Imp9 escort the MAPKs through the nuclear pores. These results suggest that β‐like importins are central mediators of stimulated nuclear translocation of signaling proteins, and therefore add a central level of regulation to stimulated transcription.
Case Report: Contribution of the Radial and Ulnar Arteries on the Blood Supply of the Thumb (LB22)
Authors: Valentin Natalia , Del Rosario Cristina , Veras Wilson
Published: June 2021

Understanding the vascular patterns of the hand and its variations is important for surgeons, radiologists and anatomists due to the constant innovations on microsurgical techniques for the traumatized hand. Even though the thumb contributes 40 percent of the overall function of the hand, its irrigation is sometimes neglected. The principal irrigation of the thumb is normally provided by the princeps pollicis artery. It has been described that this artery usually arises from the radial artery or in other occasions can arise directly from the deep palmar arch, or from a common trunk with the radilis indicis artery. However, anatomical variations to this description have been found in previous studies. In order to determine the origin of the princeps pollicis artery, 12 hands were dissected out of 6 adults formalin preserved cadavers from the Gross Anatomy Laboratory of the Universidad Central del Caribe during the fall semester of 2012‐2013. In the present study we have found 58.3% of the specimens with the princeps pollicis artery arising from the first dorsal metacarpal artery. In 85.7% of these cases there is a persistent anastomosis between the princeps pollicis and the superficial palmar arch. Additionally, the irrigation of the thumb was found as normally described in 25% of the specimens. The remaining 16.6% of the specimens were discarded. These results allow us to suggest further investigation in this topic to provide significant statistics in terms of the incidence of this anatomical variation in the population.

Grant Funding Source: Supported by: This publication was possible by grants from the National Center for Research Resources (5 G12 RR 003035‐27) and the National Institute on Minority Health and Health Disparities (8G12 MD 007583‐27) from the National Institutes of Health
Fitness and Biosensor Modules for Programmed Evolution (LB271)
Authors: Pearson Sara , Chadinha Spencer , Parrish Phoebe , Heyer Laurie , Campbell Malcolm , Poet Jeffrey , Eckdahl Todd
Published: June 2021

We are developing a process called Programmed Evolution to optimize orthogonal metabolism in E. coli. Applications of the process include the production of pharmaceuticals, biofuels, chemical commodities, and bioremediation. The system contains a gene expression device that controls production of one or more enzymes in a metabolic pathway. One Programmed Evolution cycle includes introducing genetic variation into the gene expression device using a Combinatorics Module, selecting optimal metabolism with a Fitness Module, using a Stress Response Module to mitigate damage, and measuring metabolic output with a Biosensor Module. We developed and tested Fitness Modules based on the adhE gene (for use of ethanol as an energy source) and the tetA gene (resistance to the antibiotic tetracycline). Both Fitness Modules rely on a riboswitch that is activated by binding to the product of the optimized metabolic pathway. We also incorporated the riboswitch into a Biosensor Module to measure metabolic output.
Effects of two popular weight loss programs on changes in body composition and visceral adipose tissue in women (LB296)
Authors: Coletta A , Mardock M , Lockard B , Byrd M , Simbo S , Jagim A , Kresta J , Baetge C , Jung Y , Koozehchian M , Khanna D , Kyul H , Oliver J , Greenwood M , Rasmussen C , Kreider R
Published: June 2021

The purpose of this study was to determine if different types of exercise and diet interventions promote greater changes in visceral adipose tissue (VAT). 54 sedentary women (34±8 yrs, BMI 35±6) were randomized to participate in the Curves (C) or Weight Watchers (W) weight loss programs for 16‐wks. Participants in the C program followed a 1,200 kcal/d diet for 1‐wk and 1,500 kcal/d diet for 3 wks (30:45 CHO:PRO). Subjects then ingested 2,000 kcals/d for 2‐wks (45:30 CHO:PRO) and repeated this diet while participating in the Curves with Zumba program 3‐d‐wk. Remaining subjects followed the W point‐based diet program, received weekly counseling, and were encouraged to exercise. DEXA body composition with VAT was determined at 0 & 16 wks. MANOVA revealed a significant time (p<0.001) and group x time (p=0.035) effect. Both groups lost a similar amount of weight (W ‐6.3±4, C ‐4.8±4 kg, p=0.17), fat mass (W ‐3.1±6, C ‐5.9±6 kg, p=0.13), and VAT mass (W ‐98±468, C ‐240±445 g, p=0.26). Subjects in the C group experienced greater gains in FFM (W ‐3.3±5.9. C +1.2±4.1 kg, p=0.002) and tended to lose more body fat (W ‐1.1±8, C ‐4.5±5.5 %, p=0.07). Changes in VAT mass significantly correlated with changes in weight (r=0.38), fat mass (r=0.73), FFM (r=‐0.62), and body fat (r=0.64). Results indicate that different types of diets can differentially affect changes in body composition and VAT.

Grant Funding Source: Curves International, Inc. (Waco, TX)