bluuebunny/crossref_metadata_2025 huggingface dataset:
Simultaneous measurement of the TCA cycle and respiration in isolated mitochondria and intact cells with the XF24‐3 Analyzer
Authors: Rogers George W. , Jogunoori Smitha P. , Munkholm Christiane , Neilson Andy , Ferrick David A.
Published: June 2021

Measuring mitochondrial dysfunction is increasingly important in the study of neurodegenerative and cardiovascular disease, metabolic syndrome, diabetes, cancer, and aging. Mitochondrial function is usually measured by oxygen consumption rates using respirometry methods. However, a critical aspect of mitochondrial function may be overlooked when only O2 consumption is employed: the tricarboxylic acid (TCA) or Krebs cycle, a central metabolic pathway. Measuring the TCA cycle function requires monitoring the flux of an additional analyte, such as carbon dioxide (CO2). The reported method describes simultaneous monitoring of carbon dioxide evolution rates (CDER) and oxygen consumption rates (OCR) using isolated mitochondria and intact cells, in real‐time, using the XF24‐3 Analyzer. This technology employs fluorescent sensors specific for CO2 and O2, which operate reversibly, and reveal details of mitochondrial respiration and TCA cycle activities. Results indicate that O2 consumption and CO2 evolution may be monitored simultaneously, and that data agrees with attributes of TCA cycle and mitochondrial function obtained by other methods. Differential rates of O2 and CO2 flux can be identified relative to substrate utilization and interdependency among the TCA cycle, electron transport, and oxidative phosphorylation systems. Support for this research is provided by Seahorse Bioscience.
Inhibition of cell growth by an elevated turnover number of melibiose/Na+ symport catalyzed by melibiose permease of Salmonella typhimurium
Authors: Jakkula S. Vivek , Hodkoff Alexey A. , Guan Lan
Published: June 2021

Melibiose permease of Salmonella typhimurium (MelBSt) catalyzes melibiose/cation (H+, Na+ or Li+) symport. The transport mechanism is still poorly understood. Threading model indicates that a conserved Gly117 (helix IV) is part of the Na+‐binding site. Recently, mutation analyses suggested that Gly117 plays an important role in cation binding and translocation in MelBSt. In this study, we observed that Gly117→Cys MelBSt mutant drastically inhibited cell growth in the presence of melibiose in a concentration‐dependent fashion. The mutant G117C increased Vmax value for melibiose transport by >3 fold with little changes in Km and protein expression. A spontaneous mutation Pro148→Leu was isolated from mutant G117C. Strikingly, the double mutant G117C/P148L grew at a similar rate in the absence or presence of melibiose and also largely decreased the Vmax value, with 2‐fold lower than obtained from WT. A good correlation between the Vmax values and the cell‐growth rates in the presence of melibiose indicates that the Na+/melibiose symport dissipates cell membrane potential. With a largely elevated turnover number of melibiose transport, it is likely that the cotransported Na+ may collapse the membrane potential and inhibit cell growth.

This work was supported by Texas Norman Hackerman Advanced Research Program 010674‐0034‐2009 to LG.
Dietary Histone Deacetylase Inhibitor Sulforaphane Inhibits Adipogenesis in 3T3‐L1 Pre‐adipocytes
Authors: KIM JONG SIK , Kwon Sun-Bong , Son Jung-Bin , Kim Tae-Kyun , Park Mi-Jeong , Souk Ok-Hee
Published: June 2021

In the present study, we evaluated the effects of dietary histone deacetylase inhibitor sulforaphane on cell proliferation and adipogenesis in pre‐adipocytes. Mouse 3T3‐L1 pre‐adipocytes were pre‐treated with six different phytochemicals and differentiated into adipocytes. The results showed that sulforaphane significantly decreased cell viabilities of 3T3‐L1 cells compared to other phytochemicals. Furthermore, Oil Red O staining indicated that adipogenesis was inhibited by sulforaphane in a dose‐dependent manner. To understand the molecular mechanisms involved in adipogenesis inhibition by sulforaphane, we carried out oligo DNA microarray analysis. Among down‐regulated genes, we selected five genes involved in lipid metabolism and down‐regulation of those genes was confirmed by RT‐PCR and quantitative real‐time PCR. In addition, 3T3‐L1 cells were incubated with trichostatin A, sulforaphane or 5‐aza‐deoxycytidine during adipocyte differentiation and then lipid formation was evaluated with expression level of mLeptin. Interestingly, the histone deacetylase inhibitor trichostatin A and sulforaphane suppressed mLeptin expression, whereas the DNA demethylating agent 5‐aza‐deoxycytidine increased mLeptin expression. These results indicate sulforaphane exerts anti‐obesity effects via mechanisms involving epigenic down‐regulation of adipogeneic genes.
Variation of the Radial Nerve, Ulnar Nerve and Axillary Nerve in Innervation of the Three Heads of the Triceps Brachii Muscle
Authors: Kauffman Travis , Olinger Anthony , Borman William , Fisher Cara
Published: June 2021

This study aimed to determine how frequently the radial nerve, ulnar nerve and axillary nerve provided innervation to the medial head, long head and lateral head of the triceps brachii muscle. A secondary goal of this study was to explore correlations between the distance from the point of insertion of each muscle head of the triceps brachii and where the innervating nerve enters the muscle belly. One hundred fifty‐four limbs from male and female preserved cadavers were dissected and examined during this study. The frequency of innervation by each nerve originating from the brachial plexus was recorded for the lateral head, medial head, and long head. The distance from the insertion of the triceps brachii muscle on the olecranon of the ulna to the insertion of the nerves on the muscle was measured in millimeters. The radial nerve was found providing innervation to the medial head, long head and lateral head of the triceps brachii in every limb examined. The ulnar nerve coursed through the medial head twenty times, but no branches were observed providing evidence of innervation. The axillary nerve was not observed providing innervation to any heads of the triceps brachii. The average distance from insertion of the heads of the triceps brachii on the olecranon of the ulna to the point of insertion of the innervating nerve branch on the muscle belly of the medial head, long head and lateral head was 128 mm, 183 mm and 170 mm respectively.
Receptor for Advanced Glycation End Products Functions as a Receptor for Glycosaminoglycans in Lung Metastasis
Authors: Sugahara Kazuyuki , Takahashi Jun , Mizumoto Shuji
Published: June 2021

Glycosaminoglycans (GAGs) at cell surfaces of cancer cells play roles in malignant transformation and metastasis. Highly metastatic Lewis lung carcinoma cells showed a strong expression of the chondroitin sulfate E (CS‐E) epitope consisting of the E‐disaccharide unit GlcUA‐GalNAc(4, 6‐O‐disulfate). The metastasis was efficiently inhibited in a mouse system by enzymatic removal of CS from the tumor cell surface or by pre‐injection of CS‐E in a dose‐dependent manner (Li et al., J. Biol. Chem. 2008). These findings prompted us to investigate the roleof the CS and/or GAG structure of the tumor cells in the experimental lung metastasis.

In this study to characterize CS‐E‐binding proteins in a mouse lung, an extract of a mouse lung homogenate was analyzed by affinity chromatography using a CS‐E‐immobilized column. A CS‐E‐binding putative receptor was identified as Receptor for Advanced Glycation End products (RAGE) by SDS‐PAGE followed by MALDI‐TOF‐MS of the trypsin digest of a protein band, suggesting that the E‐unit‐containing epitopes of CS chains as well as heparan sulfate chains at the tumor cell surfaces are recognized by RAGE in the lung, and are promising targets for diagnosis and therapy of malignant tumors (S. Mizumoto, J. Takahashi, and K. Sugahara, submitted).

This work was supported by a Grant‐in‐aid for Scientific Research (B) and Future Drug Discovery and Medical Care Innovation Program from MEXT (to KS).
Simplified Methods for the Study of Cell Cycle and Apoptotic Impacts of Anti‐Cancer Compounds
Authors: Tran Kimvan , Tyagarajan Kamala
Published: June 2021

Several anticancer compounds have shown to have impacts on both cell cycle and cell health with differential impacts in the timing and sequence of each of these processes. Drug development studies therefore need simple accessible simple and easy methods to study impacts on both cell cycle and multiple apoptotic markers to understand the complete dimension of action of compounds. In this study we present data from rapid simplified methods utilized to study the impact of multiple anti‐cancer and anti‐tumor compounds on cell cycle arrest and apoptosis utilizing no‐wash assays. Studies were performed with Jurkat cells with multiple anti‐cancer compounds such as gambogic acid, etoposide, celastrol, and nocodazole for differential treatment conditions and the impact on cell cycle and impact on apoptosis by annexin V binding and caspase activation was evaluated. Our results indicate that the impacts and kinetics of cell cycle versus apoptosis markers can vary with different compound treatments. Screening for both cell cycle and apoptosis impacts are thus critical in understanding the full dimensions of mode of action of compounds and can be valuable in determining optimal conditions of use.
How to Structure Small Groups for Better Performance in Medical Biochemistry
Authors: Sadik Amina , Bondarenko Vladimir , Fulop Csaba
Published: June 2021

Kolb's learning styles inventory was chosen to determine the best team settings for better small group performance in the Medical Biochemistry course.

Students were randomly grouped in the first half of the course, and then regrouped based on their learning style in the second half.

The majority of students belonged to the converging learning style followed by the assimilating style. The accommodating and diverging styles were rare.

We compared grade changes in collaborative work; concept maps (CM) and group quizzes, as well as assignments not related to collaborative work; individual quizzes (IQ). Approximately, 41% of the students had better grades in all grade categories. Although similar number of students achieved better grades in CM and IQ, only 54% of the total number of students showed improvement in both, indicating no strong correlation between the two grades. In fact, most students (79%) improved their grades of the IQ.

In teams formed by students of similar learning styles, the collaborative CM scores increased considerably; whereas their IQ scores did not seem to be affected significantly. The IQ scores of students belonging to diverse groups seemed to increase in the second part of the course. This finding might be due to the increased effort and/or familiarity with the assessment methods.
The Guanine Trap: RNA Guanine‐tract Recognition and Encaging by hnRNP F quasi‐RRM2
Authors: Neumeyer Paige , Kaufmann Phoenix , Evenson Theresa , Patynski Alexis , Schmidt Ian , Evans Elizabeth , Meissen Mitchel , Ebert Grace , Maala Nicole , Kinscher Joseph , McNally Mark
Published: June 2021

RNA splicing, the process where mRNA exons are ligated together after the introns are cut out, is required for the production of mature mRNA. Exons are the regions of mRNA that are translated into protein, and introns are noncoding regions. Alternative splicing, the process where different combinations of exons can be ligated together to generate many mRNAs from one gene, accounts for protein diversity and affects over 90 percent of human genes. Alternative splicing regulation is important because many diseases can arise if it occurs improperly. A molecular machine called the spliceosome performs RNA splicing to ligate exons, and hnRNP F (heterogeneous nuclear ribonucleoprotein) influences the spliceosome's alternative splicing decisions. hnRNP F binds to guanine‐(G) rich sequences in mRNA targets, resulting in their alternative splicing. Modeling shows a ‘cage’ formed around three G residues, which explains why hnRNP F binds G‐rich sequences. Using 3D printing technology, the Valders SMART team (Students Modeling A Research Topic) modeled hnRNP F binding via arginine 116, phenylalanine 120, and tyrosine 180. Research suggests that too much hnRNP H, a close homologue of hnRNP F, plays a role in promoting brain cancer. Understanding how hnRNP F binds G‐rich RNA to cause alternative splicing may lead to the development of therapies for genetic diseases. Supported by grants from NIH‐SEPA and NIH‐CTSI
Cathepsin S Inhibition: Reducing Pain is the Mission
Authors: Radler Shawn , Lantzsch Maddie , Finfrock Julie , Hefferan Sarah , Shimmin Bo , Currier Kristopher , Futch Liane , Ondoua Alysia
Published: June 2021

Chronic pain results from continuous inflammatory and neuropathic pain affecting 116 million Americans with conditions such as rheumatoid arthritis and multiple sclerosis. Cathepsin_S (CatS), a lysosomal cysteine protease, an important mediator of pain, is released from antigen presenting cells such as microglia. Interactions between immune cells and neurons contribute to normal pain perception. Continuous noxious stimuli cause a local ATP increase, stimulating P2x7, a microglial receptor, to release CatS. CatS cleaves fractalkine (FKN), a neuronal transmembrane protein. Active FKN binds to its receptor, CX33CRI, on microglia, releasing inflammatory mediators that activate neighboring neurons and induce the perception of pain in the affected region. Competitive inhibitors of CatS have the potential to selectively attenuate the perception of pain as evidenced in animal models. The Ironwood Ridge High School SMART Team (Students Modeling A Research Topic) has modeled interactions between CatS and competitive inhibitors using 3D printing technology. Computer imaging of this site gives critical knowledge useful towards the development of a CatS specific inhibitor to treat pain clinically. Supported by a grant from NIH‐SEPA.
Adenoviral E1A oncoprotein: The Butterfly Effect
Authors: Davidson Katie , McCarthy Aaron , McCarthy Jared , Monteverde Corrie , Monteverde Danielle , Santiago Eduardo , Galea-Martinez Sandra , Jansma Ariane
Published: June 2021

Viral infections are one of the leading causes of death in humans and are usually caused by the adenovirus. The early region protein (E1A) is a multifunctional protein expressed by the adenovirus and essential for transformation of the host cell, ultimately resulting in oncogenesis. Through E1A's interactions with the host cell's retinoblastoma protein (pRb) and CREB binding protein (CBP), E1A prevents natural cellular functions. Our focus is the interaction site between E1A and CBP/p300, specifically the transcriptional adaptor zinc finger‐2 (TAZ2) of CBP/p300. Upon interaction with the TAZ2 domain, E1A undergoes coupled folding, allowing it to bind with TAZ2, competing with, and inactivating, the transactivation domain (TAD) of the tumor suppressor p53. Through competition with p53, E1A is able to inhibit apoptosis and cell cycle arrest, thus causing the cell to divide uncontrollably. E1A has the ability to reprogram gene expression. Understanding the structure of the complex will help us to design a therapeutic that would prevent E1A from interacting with CBP, allowing normal interaction with p53 to occur. This will prevent cancer in association with adenovirus infection and help us to understand more about cancer‐causing viruses in general. The El Capitan High School SMART Team (Students Modeling A Research Topic) modeled E1A using 3D printing technology. Supported by a grant from HHMI Pre‐College Program
Modeling the Tomato Bushy Stunt Virus (TBSV), a platform for viral epitope display
Authors: Woodnight A. , Wu A. , Herperger A. , Kulkarni A. , Vacheron A. , Waizman D. , Gajiwala D. , He E. , Zhu F. , Rockwood G. , Borok G. , Shi J. , Liu J. , Pie K. , Yang L. , Xie M. , Baru N. , Wang N. , Toyonaga N. , Rhezaii O. , Hvasta R. , Rehan T. , Fisher W. , Xiang A. , Zhang Y. , Gerstin E. , Reddy V.
Published: June 2021

Several antigen display platforms are being explored to optimize the immune response. While various viruses have been studied as antigen delivery tools, the tomato bushy stunt virus (TBSV) has great potential for use as a platform due to its symmetrical structure that enhances epitope display. 180 identical subunits form a single TBSV particle that can prominently display multiple copies of an epitope. TBSV's high antigen density makes it a highly efficient means of ensuring epitope recognition, thereby increasing antibody response. TBSV is especially useful because the C‐terminus sites of its capsid protein are clearly exposed. This structure allows epitopes to be added and to maintain their original tertiary structure when bound to the viral capsid. The TBSV platform was tested by addition of modified ricin toxin to the capsid protein C‐terminus. Mice injected with the construct had increased antibody responses, indicating that the high antigen density created by modified TBSV led to increased immune response. The Canyon Crest Academy SMART Team (Students Modeling A Research Topic) modeled TBSV using 3D printing technology. This 3D model allows for more thorough analysis of TBSV's structure and fuller understanding of the characteristics that make it an attractive platform for vaccine design. Supported by a grant from the HHMI Pre‐College Program.
Retinoic Acid and Cod Liver Oil Replenishing Liver Vitamin A Stores in Vitamin A Deficient Rats
Authors: Fenlason Matthew David , Kooiman John , Standard Jacob , Becker Cassandra , Meadors Edward , Godden Kyle , Molteni Agostino , Baybutt Richard
Published: June 2021

Previously we have found that Vitamin A deficiency is associated with abnormal bone growth and development. Our studies indicated that retinoic acid (RA) appeared to be the most effective at correcting the bone pathologies associated with Vitamin A deficiency, restoring osteoblast numbers, compared to the retinyl ester form of Vitamin A found in cod liver oil (CLO). The purpose of this research was to determine which source of Vitamin A, RA or CLO, most effectively replenished Vitamin A liver stores in rats. Twelve male weanling rats were fed a retinol‐adequate (control) diet, while thirty rats were put on a retinol‐deficient diet for 9 weeks. Six rats from the control and six from the deficient group were euthanized at week 9 to verify vitamin A deficient status. The remaining 24 deficient rats were then randomly assigned to one of 4 diets, for 4 weeks: control, RA‐supplemented, CLO‐supplemented, or retinol‐deficient diet. Although the rats were pair‐fed, the vitamin A deficient rats lost weight compared to the control rats. At week 13 the experiment was terminated. The livers were analyzed for retinoid concentrations by HPLC. The results indicated that there was no significant difference between the CLO and RA treatment groups in replenishing stores. Although dietary retinoic acid was effective in partially correcting Vitamin A deficient bone pathologies, it's effectiveness was not dependent upon liver stores.
An interaction between folate and methylenetetrahydrofolate reductase alters genome‐wide DNA methylation patterns in mouse colon
Authors: Tammen Stephanie Ann , Park Lara K , Choi Sang-Woon
Published: June 2021

A common polymorphism at nucleotide 677 of the methylenetetrahydrofolate reductase (MTHFR) gene interacts with folate intake to alter genomic DNA methylation and the risk for colon cancer. We investigated the effects of reduced MTHFR and folate intake on genome‐wide DNA methylation patterns using an intestine specific MTHFR homozygous knockout (KO) mouse model fed 0 or 2ppm folate. DNA was extracted from colonic scrapings, immunoprecipitated with an anti‐5‐methlycytosine antibody and co‐hybridized with input DNA onto a DNA methylation array chip containing probes for all promoters and CpG islands. Probes with a p‐value < 0.01 and mean difference between groups > 0.5 were considered differentially methylated (DM). The contrasts of wild type and KO groups of mice fed either 0 or 2ppm folate yielded 5,613 and 6,360 DM probes respectively. Fewer DM probes (3,328) were found when analyzing the effect of folate intake in KO mice. Applying a two‐way ANOVA for MTHFR and folate interaction yields 3,161 DM probes. We conclude that MTHFR activity has more of an effect on DNA methylation patterns than folate intake. Overall, the MTHFR‐folate interaction plays a significant role in DNA methylation patterns.
The concentration of iron in Zanzibari geophagic earth is high but its in vitro bioavailability is low and it inhibits the bioavailability of dietary iron in an in vitro digestion/Caco‐2 Cell model
Authors: Seim Gretchen , Ahn Cedric , Bodis Mary , Tako Elad , Miller Dennis , Glahn Raymond , Young Sera
Published: June 2021

Geophagy, the deliberate consumption of earth, is strongly associated with Fe deficiency. Two competing hypotheses have been proposed: 1. Geophagy is a means to increase Fe status, 2. Geophagy causes Fe deficiency by inhibiting Fe absorption. Our objective was to test these using 11 geophagic earths consumed in Zanzibar, Tanzania. Fe concentrations were measured with inductively coupled plasma atomic emission spectroscopy. Fe bioavailability from earth, cooked white beans (WB), and a mixture of WB and earth (16:1 ratio) were assessed using an in vitro digestion/Caco‐2 cell model. Ferritin formation was used as an index of iron bioavailability. Median Fe concentration of the earth was 3325.7 (IQR 2439.0, 9041.3) μg/g; all were significantly higher (p<0.05, one tail) than the Fe concentration of WB (76 μg/g). Ferritin formation in cells exposed to earth was lower than in cells exposed to WB, and iron uptake responses of 9 of the 11 earth samples were not significantly different (p<0.05, two tail) from the blank, indicating no bioavailable iron. In cells with WB and earth, 4 of 11 samples had mean ferritin levels that were significantly lower (p<0.05, one tail) than the WB alone, indicating an inhibition of Fe bioavailability. In summary, although geophagic earths are high in total Fe, this Fe is not bioavailable. Further, some earth samples inhibit Fe absorption from foods. In vivo research is now warranted to confirm these observations.
Chronic oral administration of Lactobacillus casei rhamnosus 35 can modulate the gut microflora in rats
Authors: Lin Yu-Chuan , Tsai Jyun-Han , Tsao Shu-Ping , Huang Hui-Yu
Published: June 2021

Previous studies demonstrated that oral consumption of micro‐organisms can protect the gut flora. The objective of this study was to investigate the Lactobacillus casei rhamnosus 35 whether has probiotics potential on gastro intestinal (GI) protective effect in rats. Sprague Dawley rats (n=32) were divided into four groups, such as control (Con), low dose (LD), medium dose (MD) and high dose (HD). L. casei rhamnosus 35 was orally given for 8‐ week to the experimental rats. Changes in whole bodyweights and daily food intake were monitored throughout the study, and found no significant different among the groups. For the gut microflora analysis, viable cell counts of Lactobacilli, Bifidobacteria, total anaerobic bacteria counts, Escherichia coli, and Clostridium perfringens were determined from the feces and cecum of rats. In the results we found that L. casei rhamnosus 35 treatment significantly enhanced the beneficial effects on gut microflora, but suppressed the bacterial pathogens in the feces and cecum. Findings from our study suggest that L. casei rhamnosus 35 is a potential source of probiotic in rats.
Effect of long‐term high‐fat diet feeding and fasting on energy metabolic substrates utilization in resting rats
Authors: Jeon Yerim , Kim Jisu , Hwang Hyejung , Suh Heajung , Lim Kiwon
Published: June 2021

The effects of a high‐fat diet and fasting on resting energy expenditure and energy substrate utilization were examined using the calorimetry method. Eight 4‐week old rats were used for the high‐fat diet experiment. As a result of 5 weeks of control diet and high‐fat diet feeding, body weight of the high‐fat diet group tended to increase more than that in the control diet fed group, but the difference was not significant. The 24 hr respiratory exchange ratio was lower in high‐fat diet group. The control group showed significantly more carbohydrate oxidation than that of the high‐fat diet fed group. A fasting experiment was conducted using six 7‐ week old male rats. Resting metabolism rate was measured prior to fasting, and a fasting condition began from the next day for 3 days to energy metabolism. Both body weight and 24‐hour oxygen uptake decreased significantly as a result of 3‐days fasting. Total oxygen uptake in the first day decreased, and declined significantly on day 3 of fsting. The mean 24‐hour respiratory exchange ratio decreased significantly. Additionally, energy expenditure during the dark period, which is the active period for rats, decreased significantly with fasting, whereas energy expenditure during the light period did not increase by fasting. This study was supported by a Korean Research Foundation Grant funded by the Korean Government (MOEHRD) (KRF‐2009‐042‐G00058).
Comparison of analytical methods for anthocyanin quantification in berries: HPLC and pH differential methods
Authors: Lee Sang Gil , Nam Tae-Gyu , Kim Dae-Ok , Koo Sung I. , Chun Ock K.
Published: June 2021

Interest in the beneficial effects of anthocyanins on human health has resulted in increased demand for their use as food ingredients and created a need for an inexpensive and effective method to quantify the total anthocyanin contents. The pH differential method has been demonstrated to be simple, quick, and reliable for measuring the total monomeric anthocyanin content of a sample compared to HPLC method. However; results obtained by these two methods are still inconsistent. Thus, this study aimed to evaluate these two analytical methods in measuring total anthocyanin contents in berries having different anthocyanin contents and compositions. Anthocyanin fractions, obtained from the phenolic extracts of five berries (blueberry, blackberry, black currant, raspberry, and cranberry) using C18 Sep‐Pak cartridge, were used to quantitate total anthocyanin contents using pH differential and HPLC methods. In pH differential method, blueberry showed the highest anthocyanin content as 1,034 mg cyanidin‐3‐glucoside equivalent/100g dry weight, followed by blackberry (819 mg), blackcurrent (801 mg), raspberry (381 mg), and cranberry (220 mg). Results by HPLC analysis also showed similar trends but lower than those by pH differential method. In conclusion, the pH differential method is reliable and in good accordance with the data obtained by HPLC method for measuring total anthocyanin contents in berries.
Anatomy taught integration of multiple image mediums, dissection and pathology reports with donor‐cadaver‐patients
Authors: Niemeyer Matt , Delamarter Taylor , Benninger Brion
Published: June 2021

INTRODUCTION

Integrating multiple imaging modalities from a donated‐cadaver‐patient (DCP) with graded dissection and pathology recognition provides realistic clinical anatomy experience. Imaging allows students to view dissected areas and discover pathologies. Student pathology reports reinforce anatomy and radiology learning. The objective of this study was to integrate DCP imaging, dissection and pathology to provide a realistic clinical anatomy experience.

METHODS

Literature search was conducted regarding common multiple imaging modalities on DCPs during an anatomy course. Dissection and a pathology report were required.

RESULTS

Literature search revealed no studies integrating multiple image mediums on DCPs during an anatomy course. Students analyzed DCP imaging, dissected and confirmed pathology.

DISCUSSION

A vertical curriculum requires educators to integrate basic and clinical sciencesin order to develop critical thinking skills. Clinical anatomy could include radiology, surgical techniques and pathology. DCP was imaged using multiple common modalities, quality of dissection was assessed and pathology was confirmed. A DCP from this pilot study revealed bilateral popliteal aneurysms and Baker's cysts on imaging and during dissection.

CONCLUSION

This pilot study integrated radiology, dissection and pathology from DCP's to provide a realistic clinical anatomy experience.
Immunomodulating effects of glycoprotein fraction isolated from rice bran on a Raw 264.7 macrophage cell line
Authors: Park Ho-young , Choi Heedon , Yu Areum , Choi Inwook , Kim Yoonsook
Published: June 2021

The immunomodulatory activities of glycoprotein isolated from rice (Oryza sativa) bran were assessed. The glycoprotein fraction from rice bran (GFRB) prepared by 80% ammonium sulfate treatment against aqueous extract of rice bran contained 65.7% protein and 7.7% neutral sugar and showed 4.9% extraction yield. The GFRB consisted of five glycoproteins with different molecular weights and remarkably improved production of nitric oxide (NO) in murine macrophage cell, up to approximately 10‐fold compared to the control at 100 ug/mL concentration. Cytokines such as interleukin‐1beta (IL‐1β), IL‐6, IL‐10 and tumor necrosis factor‐alpha (TNF‐α) were secreted from raw 264.7 cells at levels of 224.5±4.0 pg/mL, 524.3±4.8 pg/mL, 133.0±9.5 pg/mL and 28602.0±12.5 pg/mL, respectively, at 50 ug/mL of GFRB concentration, and these levels were higher than the levels measured in the normal control. And the GFRB showed no cytotoxicity. These results demonstrated that the GFRB has potential as a new functional food material that evidences immunomodulatory activity.
Anatomy to Acoustics: The Mechanism of Humpback Whale Sound Production
Authors: Adam Olivier , Reidenberg Joy S , Laitman Jeffrey
Published: June 2021

Humpback whales emit complex songs comprised of successive sound units with a fundamental frequency and harmonics. It is unknown how these calls are produced, given that the respiratory tract is closed (i.e., no egress of air from the nose or mouth) during underwater vocalizations. Unlike open system of terrestrial mammals, the whale vocal tract is subject to flow cessation as pressure builds. Prior studies on cetacean laryngeal anatomy (Reidenberg and Laitman, 2007) showed a communication for airflow between the trachea and an expandable ventral diverticulum called the laryngeal sac. We propose a theoretical model of humpback whale sound production based on the source‐filter theory that accounts for a closed system constrained by the dimensions of the trachea, the nasopharynx, and the laryngeal sac. We used recordings of humpback whales from the Indian Ocean (provided by Cetamada Association, Madagascar). Results show a relationship between time (duration) and spectral features (fundamental frequencies) of these calls. As this is a closed system, duration is dependent upon laryngeal sac pliability, while frequency is determined by vocal tract length. Our investigation is a first step towards defining individual acoustic signatures. Future studies will assess whether predicted durations and frequencies, based upon known vocal tract measures, fall within the range of actual recorded sounds.
The relationship between dietary intake and allergic disease in Korean adolescents: The Fourth Korea National Health and Nutrition Examination Survey (2007–2009)
Authors: Kim Hyesook , Kim Ki-Nam , Hur Jinhee , Kim Soyeon , Kim Yuri , Oh Se-Young , Kang Myung-Hee , Chang Namsoo
Published: June 2021

We investigated the relationship between dietary factors and allergic disease (AD) in Korean adolescents, aged 12–18 years, using the data of the Korea National Health and Nutrition Examination Survey (KNHANES IV, 2007–2009). From a total of 2,485 subjects, 112 had asthma, 269 atopy, and 354 either asthma or atopy. Adolescents with AD were found to consume more total energy. After adjusting for potential confounders, adolescents with AD were found to consume more energy from carbohydrates than those without. Among micronutrients iron intake was higher in subjects with AD than those without. When we further examined gender difference in the relationship between nutrient intake and AD, boys with AD consumed more energy from carbohydrates, and iron. Girls on the other hand, the relationship between nutrient intake and AD was not observed except for total energy intake. Further studies are needed to identify specific dietary patterns that are associated with AD in South Korean adolescents.

This work was supported by the research grant (#911047‐1) from the projects for the ‘Globalization of Korean Foods’ and the Brain Korea 21 project.
Rat models for the study of cholesterol disequilibria
Authors: McCoy Aaron Michael
Published: June 2021

Cholesterol homeostasis has been correlated to several human disease pathologies including obesity, diabetes, atherosclerosis, Alzheimer's disease and Huntington's disease. To develop a better understanding of the mechanisms of cholesterol regulation we have created rats with manipulations in the genes Apoe, Ldlr and Lep. These mutants were created by injection of zinc finger nucleases (ZFN) into one‐cell Sprague‐Dawley rat embryos. Apoe KO animals demonstrate a 4.5 fold increase in total serum cholesterol concentration at 10 weeks relative to their wild type littermates. Rats deficient in Ldlr demonstrate a 4–8 fold increase in total serum cholesterol and a slight increase (~10%) in weight gain at 18 weeks relative to WT. Lep deficient rats demonstrate a 2 fold increase in body weight at 18 weeks relative to WT and a nearly a 4‐fold increase total triglyceride blood concentrations post 16hr fast. Additionally, we performed a glucose tolerance assay in Lep deficient and WT animals at 5 and 10 week time points. Lep deficient animals consistently demonstrated increased concentrations of glucose in the blood, indicating impaired glucose tolerance in the absence of functional Lep.
Effects of cayenne pepper on metabolic dysregulation
Authors: Plumlee Sonya Deva , Baca Kyrie , Hong Mee Young
Published: June 2021

Cayenne pepper is a strong anti‐inflammatory agent has been associated with increased metabolism. What is unclear is if dietary consumption of cayenne pepper attenuates the risk factors associated with metabolic dysregulation, namely, weight gain, lipid profile, glucose control and inflammation. Forty 21‐day old Sprague‐Dawley rats were separated into four weight‐matched groups (n=10). A 2 (diet) × 2 (treatment) factorial‐design was performed using an atherogenic diet with or without a cayenne component and with or without a Dextran sodium sulfate (DSS) inflammation inducing agent treatment for four weeks. Experimental cayenne powder was determined to contain 82,879 heat units and the following percent progression was performed with cayenne groups, 3 days 0.6%, 4 days 1.2%, 3 days 1.8% and 15 days at 2.9%. Cayenne pepper treatment groups showed significant decreases in serum tryglycerides (p =0.032), total cholesterol (p=0.039), LDL cholesterol (p=0.032), final body weight (p=0.046), weight gain (p=0.023), final epididymal fat weight (p=0.038), C‐reactive protein (p=0.044) and significant increases in HDL cholesterol (= 0.046). Cayenne pepper consumption did not significantly decrease serum glucose. These results support the use of dietary cayenne pepper consumption for the attenuation of the factors associated with metabolic dysregulation. Supported by SDSU UGP and NUTR 302L class.
Prevalence of multimorbidity among U.S. adults: data from the National Health and Nutrition Examination Survey
Authors: Staimez Lisa Rachel , Wei Melissa Y , Ramakrishnan Usha , Narayan K.M. Venkat , Saydah Sharon
Published: June 2021

Background

Chronic diseases have common modifiable risk factors, among them poor nutrition. Multimorbidity, the presence of two or more chronic diseases in an individual, and its associated risk factors have not been well characterized.

Methods

We analyzed data from non‐pregnant adults = 20 years in the National Health and Nutrition Examination Survey, 2003–2008. Cardiovascular disease (CVD), chronic obstructive pulmonary disease (COPD) and diabetes were based on self‐report. Chronic kidney disease (CKD) was computed using clinical parameters. Results were age‐standardized to the 2000 US census population.

Results

Of 13446 individuals with data on CVD, COPD, CKD and diabetes, 8.8% (standard error (SE), 0.3) had multimorbidity. Specifically, 6.6% (SE 0.2) had two diseases, 2.0% (SE 0.2) had three diseases, and 0.4% (SE 0.1) had four diseases, while 19.4% (SE 0.4) had one disease. CKD was most prevalent (15.5%, SE 0.4), then CVD (8.8%, SE 0.4), diabetes (7.8%, SE 0.4) and COPD (7.6%, SE 0.4). Multimorbidity was highest among adults >60 years (25.7%, SE 1.0), then 40–59 years (6.5%, SE 0.5) and 20–39 years (1.2%, SE 0.2). Multimorbidity was similar across sex.

Conclusions

One‐third of U.S. adults have at least one chronic disease, and about nine percent have two or more chronic diseases. Targeting common risk factors such as nutrition may have high impact on the prevention of multimorbidity.
Triacsin C Suppresses Non‐Esterified Fatty Acids (NEFA) and Increases Nitric Oxide (NO) Synthesis
Authors: Blakeman Nina Ruth , Trepanier Marc-Olivier , Bazinet Richard P. , Weis Margaret T.
Published: June 2021

Endothelial dysfunction is characterized by decreased vascular NO availability. Elevated NEFA decreases eNOS activity. In cultured cells, we found that fatty acyl CoA synthase (FACS) inhibitor Triacsin C (TC) interrupted eNOS palmitoylation, increasing eNOS activity but not changing vascular‐active eicosanoids. Hypothesis: TC mitigates endothelial dysfunction by increasing NO and decreasing NEFA. In this study, intravascular NO synthesis was measured by electrochemistry in the ischemic hind limb with heparinized rat model in a time course design. The post‐ischemic NO was significantly elevated in TC (100 μg/kg)‐treated animals than controls. NOS inhibitor treatments in this model implicate a role of eNOS, but not iNOS. Acute hyperlipidemia was induced by a bolus dose of Intralipid®(1 ml) to increase NEFA. Pre‐treatment with TC had no effect on total plasma lipids or triglycerides, either before or after Intralipid®. TC reduced baseline plasma NEFA from 240 ± 26.8 to 147 ± 14.3 μg/ml (p=0.038). Fifteen minutes after Intralipid®, NEFA rose to 1461 ± 130 μg/ml (control), which was blunted by TC to 393 ± 2.55 μg/ml (p=0.0079). By 85 min., the difference had subsided (194 ± 10.2 control vs. 281 ± 74.8 μg/ml TC; p=0.74). The TC effect was not uniform across all fatty acid species measured. These data show that TC increases post‐ischemic eNOS activity and blunted plasma NEFA induced by acute hyperlipidemia.
Deficiency of secreted cryptdin‐4 detected in a mouse model of Crohn fs disease using a new sandwich ELISA
Authors: Nakamura Kiminori , Kuroishi Aiko , Kono Mai , Kobayashi-Sakamoto Michiyo , Ayabe Tokiyoshi
Published: June 2021

Paneth cells in the base of small intestinal crypts secrete α‐defensins in response to bacteria, contribute to innate immunity, and shape the composition of intestinal microbiota. Among mouse α‐defensins, cryptdin‐4 (Crp4) is the most potent microbicide. However, a lack of sensitive assays to quantify α‐defensins released into the intestinal lumen has limited efforts to establish a relationship between secreted α‐defensin and diseases. Here we establish a highly sensitive sandwich ELISA, and analyze secreted Crp4 in feces from a mouse model of Crohn's disease. SD rats were immunized with native Crp4, and monoclonal antibodies recognizing different epitopes were identified and paired to optimize ELISA conditions. To measure levels of secreted Crp4, feces were collected from BALB/c mice and IL10 null, Crohn's disease model mice, and fecal proteins were extracted. The established ELISA was specific and highly sensitive with a Crp4 detection range of 120~1600 pg/ml. The Crp4 concentration measured in feces of normal mice was 20.4 ± 5.1 ng/ml (n=5). In contrast, feces from IL10 null mice contained 8.7 ± 5.0 ng/ml of Crp4 (n=5). Secreted Crp4 was significantly reduced in IL10 null mice. Quantifying secreted α‐defensin in feces enable us to know luminal α‐defensin levels for the first time, and contributes to a deeper understanding of diverse diseases such as inflammatory bowel disease, infectious disease, and obesity.
Universal Adapter for Telecytopathology
Authors: Markovic Olivera , Markovic Nenad
Published: June 2021

We present a prototype of a Universal Adapter to connect “any” microscope with “any” cellphone camera. It is intended to be used as an accessory medical device for wireless and digital telecytopathology – mass cervical cancer screening in particular.

For the first time, at EB 2004, we presented our proprietary, biomarker‐based MarkPap® platform technology for detection of abnormal cells on cervical specimens. At the subsequent meetings we followed with further developments, from simple manual MarkPap® Test through semi‐automated processing, self‐home specimen's collection, to digital and mobile telecytopathology. The universal adapter has two platforms, one for the microscope and the other for cellphone cameras and has a capability for 4D movements to achieve the best optical conditions for image capturing.

In case of mass cervical cancer screening, the health care providers (even low trained technicians) at the point of care processes the specimen with the customer‐friendly MarkPap® Kit, searches for biomarker‐highlighted abnormal cells, captures those images with cellphone camera using the adapter and forwards only those images for distant diagnosis. The result can be returned while women are still on the premises.

This work is a part of our current project for development of simple, low‐cost medical devices for low‐resource areas worldwide.

This work was supported by BioSciCon, Inc., Rockville, Maryland.
Anti‐inflammatory Effects of Peanut Skin Extracts on COX‐2 in Raw 264.7 Cells
Authors: Lewis Wanida , Dean Lisa , Harris Gabriel Keith
Published: June 2021

Peanut skins have been regarded as a low economic waste by‐product of the peanut industry but they contain high levels of bioactive compounds including catechins and procyanidins, which could benefit human health. This is the first report that investigated the anti‐inflammatory effects of peanut skin extracts (PSE) in vitro. Defatted peanut skins were extracted using aqueous solvent mixtures (acetone/water and ethanol/water) and dried to powders. Antioxidant activity was determined by Hydrophilic Oxygen Radical Absorbance Capacity (H‐ORAC) and Total Phenol assays. PSE extracted with acetone/water had higher H‐ORAC and total phenolic values (3060 μmol Trolox/100 g and 290 mg GAE/g) than PSE extracted with ethanol/water (2620 μmol Trolox/100 g and 250 mg GAE/g, respectively). Raw 264.7 mouse cells were treated with three concentrations of PSE (1, 2.5 and 5% (w/v)) and induced with an inflammatory marker, lipopolysaccharide (LPS) for 16 hours. Prostaglandin PGE2 was measured by enzyme linked immunosorbant assay (ELISA). Increasing concentrations of PSE induced with LPS demonstrated a decrease in PGE2 activity. PSE extracted with acetone or ethanol treatments of 5% showed significantly decreased concentrations of PGE2 activity compared to the negative control (p<0.05). Western blotting analysis showed an inhibition in COX‐2 expression as PSE levels increased suggesting that PSE's inhibition of PGE2 inhibits COX‐2. This study highlights the need to investigate the relationship between antioxidant and anti‐inflammatory properties of a waste by‐product that has the potential use as anti‐inflammatory agent.
Biological characterization of THRX‐956485, a peripherally‐selective serotonin transport reuptake inhibitor
Authors: McConn Donavon , Tsuruda Pamela , McNamara Alexander , Hegde Sharath , Stangeland Eric
Published: June 2021

Peripheral serotonin has been implicated in many therapeutic indications including thrombosis, pulmonary hypertension and osteoporosis. A peripheral selective serotonin reuptake inhibitor (SSRI) without CNS activity could provide a therapeutic index exceeding standard SSRIs. We describe a novel compound that selectively inhibits peripheral serotonin reuptake.

THRX‐956485 is a potent (hSERT uptake IC50/binding Ki = 2.5nM/1.0nM) and selective SERT inhibitor in vitro and is >2500‐ fold selective over other reuptake transporters and serotonin receptors. In rat PK studies (single 5 mg/kg PO), or 14‐day osmotic mini‐pump infusions (0.3, 0.7 and 1.6 mg/kg/day), THRX‐956485 showed pronounced peripheral selectivity compared to a CNS‐penetrant SSRI, fluoxetine. Based on unbound CNS vs. plasma exposures and measured SERT occupancy ex vivo, THRX‐ 956485 was >20‐fold peripherally selective (CSF:unbound plasma concentration ratios varied from 1–5%). Additionally, THRX‐956485 demonstrated a dose‐dependent reduction in blood serotonin concentrations in rats. Following 14‐day infusions, blood serotonin concentrations decreased from 94 ng/mL (vehicle) to 43, 25, and 11 ng/mL at 0.3, 0.7 and 1.6 mg/kg/day, respectively. These findings are consistent with inhibition of platelet serotonin uptake, and suggest that THRX‐956485 is a peripherally selective SSRI.
A novel peripheral CB1 antagonist in enhancing energy expenditure
Authors: Hung Ming-Shiu , Yeh Yen-Nan , Hsiao Wen-Chi , Shia Kak-Shan
Published: June 2021

Cannabinoid receptor 1 (CB1), a G protein‐coupled receptor, regulates appetite and metabolic pathways via both central and peripheral mechanisms, and has become a potential therapeutic target for treating metabolic diseases. To limit the psychological side effects resulted from the central action of CB1 receptors, identification of peripheral CB1 antagonists are being actively pursued. A novel alkynylthiophene series of CB1 antagonists identified demonstrated low brain/plasma (B/P) ratio, and compound 1 is a representative of this series. Compound 1 is an orally active inverse agonist. Chronic blockade of CB1 ameliorated obesity‐associated insulin resistance, and upregulated genes for energy expenditure, like uncoupling protein 1 (UCP1) and b2‐adrenergic receptor (b2‐AR) in visceral adipose tissue in DIO mice. These results suggest peripheral acting CB1 antagonists may contribute to energy expenditure by transdifferentiation of white adipose tissue to brown adipose tissue‐like property.
Complete artificial saliva alters expression of proinflammatory cytokines in human dermal fibroblasts
Authors: MALPASS GLORIA , PRASAD GADDAMANUGU , HOWLETT ALLYN
Published: June 2021

Complete artificial saliva (CAS) is a saliva substitute often used as a solvent control/vehicle for test reagents. In cultured normal human dermal fibroblasts incubated for 5 hours with CAS, gene expression for proinflammatory cytokines including interleukin 8 (IL8) and interleukin 1á (IL1á) was increased. Expression of vascular cell adhesion molecule 1 (VCAM1), a gene upregulated by certain proinflammatory cytokines including IL1á, was also increased. Furthermore, cytometric bead array assays indicate that CAS increased the release of proinflammatory cytokines including IL8 into the cell culture media. These results suggest that constituents of CAS may induce a proinflammatory response in cultured human dermal fibroblasts. Therefore, it is important to carefully evaluate/consider the “vehicle effects” of CAS and its constituents in oral/dental research. Funded by RJ Reynolds Golberg Postdoctoral Fellowship to GEM.