bluuebunny/crossref_metadata_2025 huggingface dataset:
Eps8 facilitates phagocytosis by increasing TLR4‐MYD88 interaction in LPS‐stimulated macrophages
Authors: Leu Tzeng-Horng , Chen Yen-Jen , Maa Ming-Chei
Published: June 2021

Toll‐like receptors (TLRs) are crucial in macrophage phagocytosis, which is pivotal in host innate immune response. However, the detailed mechanism is not fully defined. Here, we demonstrated the induction of Src and Eps8 in LPS‐treated macrophages was TLR4‐ and MyD88‐dependent, and their attenuation reduced LPS‐promoted phagocytosis. Confocal microscopy indicated the colocalization of Eps8 and TLR4 in the cytosol and at the phagosome. Consistently, both Eps8 and TLR4 were present in the same immunocomplex regardless of LPS stimulation. Inhibition of this complex formation by eps8 siRNA or overexpression of PH‐truncated Eps8 (i.e. 261‐p97Eps8) decreased LPS‐induced TLR4‐MyD88 interaction and the following activation of Src, FAK, and p38 MAPK. Importantly, attenuation of Eps8 impaired bacterium killing ability of macrophages. Thus, Eps8 is a key regulator of the LPS‐stimulated TLR4‐MyD88 interaction and contributes to macrophage phagocytosis. This work was supported by the National Science Council (NSC100‐2325‐B‐006‐011‐)
Spinasterol‐glucose inhibits IFN‐γ/TNF‐α‐induced expression of TARC/CCL17 via blocking NF‐κB and STAT1 activation in human HaCaT keratinocytes
Authors: Jung MiRa , Lee Tae Hoon , Kim Jiyoung
Published: June 2021

TARC/CCL17 is a Th2‐type chemokine and is implicated in the development of Th2‐skewed allergic diseases. TARC/CCL17 expression is induced by pro‐inflammatory cytokines such as IFN‐γ and TNF‐α. A phytosterol, 3‐O‐β‐D‐glucopyanosylspinasterol (spinasterol‐Glc), was isolated from Stewartia koreana leaves, and was previously shown to inhibit the production of NO and pro‐inflammatory cytokines in macrophage cells. In the present study, we investigated the effect of Spinasterol‐Glc on production of TARC/CCL17 induced by IFN‐γ and TNF‐α in HaCaT keratinocytes. Pretreatment of HaCaT cells with spinasterol‐Glc inhibited the mRNA and protein expression of TARC/CCL17 induced by IFN‐γ/TNF‐α in a dose‐dependent manner. Spinasterol‐ Glc suppressed IFN‐γ/TNF‐α‐induced activation of NF‐κB and STAT1. Spinastrol‐Glc also inhibited phosphorylation of IκB‐α and IKKα/β as well as nuclear translocation of NF‐κB. We demonstrated that spinasterol‐Glc reduced IFN‐γ/TNF‐α‐induced transcription of the NF‐κB‐ and the GAS‐ minimal promoters. In addition, spinasterl‐Glc suppressed the DNA binding of NF‐κB and STAT1 to its cognate consensus site. These results suggest that spinasterol‐Glc has effective inhibitory effects on production of TARC/CCL17 via inhibition of NF‐κB as well as STAT activation, and may be used as a potential anti‐inflammatory agent for the prevention and treatment of skin inflammatory diseases.
Anti‐Mitotic Chemotherapy Activates CREB and MAPK Associated Survival Signaling in B Cell Lymphoma
Authors: Gravis Demetrius Peter , Wolf Katharine , Salas Nico , Meltzer David , Sullivan Kellen , Dickson Molly
Published: June 2021

Cyclic AMP responsive‐element binding protein (CREB) is a transcription factor linked to proliferation and survival of many cell types, including B lymphocytes and B cell lymphomas. Cancer cells can increase resistance to chemotherapy via activating survival signaling pathways, and we investigated whether CREB activating pathways are associated with chemoresistance signaling in B lymphomas. B lymphoma cell lines were treated with anti‐mitotic drugs including etoposide, which induces DNA damage, and colchicine, which promotes microtubule depolymerization. We observed that etoposide and colchicine promoted CREB phosphorylation via signaling pathways involving protein kinase C (PKC) and mitogen activated protein kinases (MAPK). We next tested whether the CREB activating PKC and MAPK pathways influenced chemosensitivity. PKC and MAPK inhibitors, used either singly or in combination with colchicine or etoposide, enhanced B lymphoma growth arrest and cell death. Our results support the continuing study of combination therapies pairing traditional anti‐neoplastic drugs with selective signal transduction inhibitors to combat chemoresistance and increase chemotherapeutic efficacy.
IDENTIFICATION OF CYTOCHROME P450 3A4 PRESENT IN CHO CELLS TREATED WITH A CITRUS HYBRID
Authors: Rodriguez Robert , Santos Santori Lizette
Published: June 2021

Orangelo or best known as ‘Chironja’ is a hybrid citrus fruit between orange and grapefruit. Previous investigations agree that different compounds found in citrus fruits like orange and grapefruit inhibit, completely or partially, the activity of different isoenzymes in the cytochrome P450 system. These compounds are ascorbic acid, flavonoids, limonoids, furocoumarins and other related complexes. The effect of this hybrid in the P450 system of Chinese Hamster Ovarian Cells, especially in CYP3A4 isoenzyme was studied. Extraction from the fruit was performed by squeezing three chironjas. CHO cells were treated for 48 hours with the chironja's extract. After this procedure the luminescence of each sample was determined using an appropriate luminescent compound. Intensity was measured at 450 nm using a spectrofluorometer. Results were: 0.827 pmol d‐luciferin/ pmol/min., 2.44 pmol d‐luciferin/pmol/min., and 1.26 pmol d‐luciferin/pmol/min., control, organic extract and treated cells, respectively. An increase on enzyme activity was observed. Further studies are being conducted in order to establish the effect of this extract on the isoenzyme.
Anatomical and clinical aspects regarding renal lymphatic drainage. A dissection study
Authors: Daescu Ecaterina , Alexa Aurora , Baderca Flavia , Zahoi Delia Elena
Published: June 2021

The study aims to analyse the topography of the renal lymphatics and their interrelations with the renal pedicle. 30 unfixed pieces sampled in blocks with the large abdominal vessels were used for dissection. The appearance and disposition of the collecting lymph trunks were analysed in relation to the arterial plane. Collecting lymph trunks were identified bilaterally, both anteriorly and posteriorly, overlapped with the renal nerve plexus. One or two middle collecting trunks were identified only in the renal pedicle, but they ended in the middle portion of the pedicle, draining into the posterior or anterior collecting trunks, not directly into the lumbar lymph nodes. On leaving the renal hilum, the anterior collecting lymph trunks (2–4) followed divergent courses, while the posterior lymph trunks (2–5) followed a parallel course mostly. In the middle portion of the renal pedicle, both types of trunks converge into a network whose terminal branches drain into the lumbar lymph nodes. The anterior and posterior collectors were interconnected through many branches travelling above the renal pedicles. The disposition of the collecting lymph trunks is closely related to the renal vascular morphology that changes the type of the lymph drainage of a kidney. Such aspects are useful to know not only for anatomical, but also clinical and surgical purposes (in lymphadenectomies).
Transgenic Expression and Purification of Myosin Isoforms Using the Drosophila melanogaster Indirect Flight Muscle System
Authors: Caldwell James Tore , Melkani Girish C. , Huxford Tom , Bernstein Sanford I.
Published: June 2021

We report a novel method for the generation of extremely pure transgenic myosin to carry out crystallization screening for x‐ray crystallography. We are optimizing a novel method for the use of genetically engineered Drosophila melanogaster to express histidine‐tagged recombinant muscle myosin isoforms. This method relies on several facts. First, that the Drosophila genome contains only one gene encoding all skeletal muscle myosin heavy chain isoforms. Second, that wild‐type or mutagenized versions of the gene can then be inserted into a specific location in the Drosophila genome. Third, that the Mhc10 fly line has endogenous myosin knocked out in the indirect flight muscles. Fourth, that the Actin88F promoter facilitates high‐level expression of accessible myosin in the thoracic indirect flight muscles. Finally, that cost of fruit fly colony propagation is relatively low. Using these advantages we devised a method for expressing and purifying two isoforms of recombinant histidine‐tagged myosin from an engineered fly strain. The myosin shows ATPase activity and is of sufficient purity and homogeneity for crystallization. This technique may prove useful for the expression and isolation of mutant myosins linked with skeletal muscle diseases and cardiomyopathies for their biochemical and structural characterization. Supported by NIH R01 GM032443 to SIB.
KCS1 deletion in Saccharomyces cerevisiae leads to a defect in translocation of autophagic proteins and reduces autophagosome formation
Authors: Chen Michael Po-Hao , Taylor Robert , Chou Chia-Ching , Jin Shengkan
Published: June 2021

Inositol phosphates are implicated in the regulation of autophagy; however, the exact role of each inositol phosphate species is unclear. In this study, we systematically analyzed the highly conserved inositol polyphosphate synthesis pathway in S. cerevisiae for its role in regulating autophagy. Using yeast mutants that harbored a deletion in each of the genes within the inositol polyphosphate synthesis pathway, we found that deletion of KCS1, and to a lesser degree IPK2, led to defect in autophagy. KCS1 encodes an inositol hexakisphosphate/heptakisposphate kinase that synthesizes 5‐IP7 and IP8; and IPK2 encodes an inositol polyphosphate multikinase required for synthesis of IP4 and IP5. We characterized the kcs1Δ mutant strain in detail. The kcs1Δ yeast exhibited reduced autophagic flux, which might be caused by both the reduction in autophagosome number and autophagosome size as observed under nitrogen starvation. The autophagy defect in kcs1Δ strain was associated with mislocalization of pre‐autophagosomal structure (PAS), a defect in Atg18p release from the vacuole membrane and mislocalization of PtdIns(3)P under nitrogen deprivation conditions. Interestingly, formation of autophagosome‐like vesicles was commonly observed to originate from the plasma membrane in the kcs1Δ deletion strain. Our results indicate that lack of KCS1 interferes with proper localization of some autophagic proteins, leads to reduction of autophagosome formation, and causes the formation of autophagosome‐like structure in abnormal subcellular locations.

Michael Po‐Hao Chen and Robert Taylor Jr. both contributed equally to the work.
Characterization of the Radial Artery for the Purposes of Catheterization
Authors: Wessel Erich Matthew , Hessel Kara , Olinger Anthony B.
Published: June 2021

Transradial coronary artery intervention is a safe alternative to traditional catheterization techniques that utilize the femoral or brachial arteries. There is no consensus in the literature as to which point of the radial artery (RA) is the safest to introduce a catheter. The purpose of this study was to measure the diameter, tortuosity and branching patterns of the distal RA with respect to the radial styloid process (SP). 93 cadaveric RAs (46 male, 47 female) were dissected and measured. The diameter of the RA was measured at 2 cm increments starting at the SP and moving proximally along lateral edge of the radius. The location and degree of tortuosity of the artery were recorded if >35□. Lastly, branches of the RA were recorded with respect to their distance from the SP. In males, we found the average diameter of the RA at the SP was significantly larger than the diameter at 6 cm and 10 cm. In males, we observed typical RA branches at an average of 1.72 cm from the SP and tortuosity of the RA >35□ at 1.65 cm from the SP. In females, there were no significant changes in the diameter of the RA along its course. In females, branches and tortuosity >35□ occurred on average at 1.98 cm and 2.93 cm from the SP respectively. Our data suggests that catheters could be safely inserted at distances greater than 3 cm from the SP to avoid excessive tortuosity or branches, and greater accessibility may be found 6 cm proximal from the SP in males.
Sphingolipid metabolism in Xenopus laevis oocyte maturation and apoptotic death
Authors: Petcoff Douglas W. , Bulow Aviva A. , Hostetter Trevor D. , Stith Bradley J.
Published: June 2021

Physiological maturation of Xenopus laevis oocytes is brought about by progesterone (PG). The action of PG involves an increase in ceramide and can be mimicked by injection of ceramide or treatment with sphingomyelinase (SMase)(Strum et al. 1995). Whereas prophase‐arrested Xenopus oocytes can be maintained in culture for several days, matured oocytes or eggs (both MII‐arrested) die rather quickly. The apoptotic death of maturing Xenopus oocytes has recently been shown to correlate with increased phosphorylation of Bad on residue S128 (Du Pasquier et al. 2011). We have initiated a study to determine the relationship between sphingolipid metabolism, Bad S128 phosphorylation, and cell death in Xenopus oocytes. We have found that treating cells with SMase triggers maturation and death in a manner that is similar to the effects of PG treatment. These events correlate to an increase in phosphorylation of Bad S128 as evidenced by western blotting with an anti‐S128 Bad phosphospecific antibody. Phosphorylation of this residue is slowed by including the JNK inhibitor SP600125 in the assay. To assess the post‐mitochondrial phase of apoptosis, we have used a microinjected near‐infrared dye caspase substrate to show that treatment with ceramide increases the activity of caspase 3. In characterizing the roles of SM pathways, JNK activity, and Bad S128 phosphorylation state in the biochemistry of maturing oocytes, we are currently using both genetic and pharmacologic approaches to teasing apart maturation and death. For example, using cycloheximide we have shown that both PG and SMase‐induced maturation require protein synthesis, whereas phosphorylation of Bad S128 does not. Results of experiments currently under way using genetic manipulation of JNK and sphingosine kinase 1 (SK1) will also be reported.
RTK Phosphorylation
Authors: Schmidt Jane , Schmidt Jane
Published: June 2021

A homogeneous multiplex immunoassay for RTK phosphorylation state that is adaptable to HTS

Jane Schmidt, Ph.D., Judy Haning, Shaya Anderson, Richard Fuerstenberg. R&D Systems, Inc. Minneapolis, MN

Receptor Tyrosine Kinases (RTKs) are widely expressed transmembrane protein receptors for various extracellular signaling molecules. Binding of a specific ligand leads to phosphorylation at specific sites in the cytoplasmic tail which triggers further signal transduction processes. RTKs play critical roles in the regulation of developmental processes including cell survival, proliferation, and motility, which are involved in cancer formation when not properly regulated. For these reasons, RTKs have become popular therapeutic targets for drug development. The Human RTK VersaMAP Magnetic Multiplex System kits are bead‐based multiplex immunoassays for the analysis of RTK phosphorylation. Due to the homogeneous assay format and small sample volume, these assays can be readily adapted for high‐throughput screening with minimal manual intervention from cell treatment to data collection. To demonstrate this, several different cell lines were grown in 96‐well plates, treated with various stimulants or inhibitors, and lysed in the same plate. Diluted lysates were transferred to the assay plate for analysis of RTK phosphorylation by Human RTK VersaMAP assays in two multiplexes of 25 each. Using these assays, relative phosphorylation states were successfully determined for the various cells and treatments.
Student Learning Outcomes and Attitudes when Biotechnology Lab Partners are of Different Academic Levels
Authors: Witherow D. Scott , Miller Heather , Carson Susan
Published: June 2021

Courses teaching fundamental concepts and methodologies in biochemistry and molecular biology can be beneficial for both undergraduate and graduate students alike. Should these courses be taught differently and separately to students of different levels? Anecdotal evidence has shown that students prefer pairing with someone of the same academic level. However, retention of main ideas in a peer learning environment has been shown to be greater when partners have dissimilar abilities. In our “Manipulation and Expression of Recombinant DNA” course, which can be taught as an advanced Biochemistry/Molecular Biology course, we tested the hypothesis that there will be enhanced student learning when lab partners are of different academic levels. Learning outcomes were met by both undergraduate and graduate students, regardless of pairing. Average undergraduate grades on every assessment method increased when paired with graduate students. Many of the average graduate student grades also increased modestly when paired with an undergraduate. Attitudes toward working with partners dramatically shifted. The work presented here suggests that offering dual‐level courses benefit students of all levels. This format is useful for institutions that wish to offer “boutique” courses where student enrollment may be low, but specialized equipment and faculty expertise is needed.

This work was supported by the NCSU Biotechnology Program.
Transthyretin Stabilizers: Preventing Amyloidosis
Authors: Arakawa Reina , Bang Jakyung , Rotondo Michael , Saenz Andres , Sherman Noah , Harrold Eric , O'Dea Conor , Yan Vincent , Rokous Judy South , Connors Lawreen H.
Published: June 2021

Amyloidosis, a disease caused by aggregation of misfolded protein, affects millions of people. Deposition of aggregates interferes with normal function of organs and is sometimes caused by mutant forms of transthyretin (TTR), a protein found in blood and cerebrospinal fluid. Wild‐type TTR is a homotetramer that carries thyroxine (T4), a hormone that regulates metabolism. The tetramer arrangement forms a central channel which contains binding sites for T4. One point mutation responsible for V122I is amyloidogenic and common in African Americans (4% allele frequency). This substitution likely causes destabilization of TTR's native conformation, promoting dissociation to partially unfolded monomers. The monomers aggregate to form insoluble amyloid fibrils that are deposited in the heart. Valine and isoleucine, both hydrophobic amino acids, differ only by one methyl group; how this small alteration causes cardiac amyloidosis is unclear. T4 analog diflunisal, previously used to treat the disease, inhibits the vital enzyme cyclooxygenase. The Governor's Academy SMART (Students Modeling a Research Topic) team designed a model using 3D printing technology to highlight important structural features of TTR. A thorough understanding of the T4 binding site within TTR will allow design of effective kinetic stabilizers. Supported by a grant from NIH‐SEPA.
Designing and Implementing Team‐Based Learning Exercises in a Medical Biochemistry and Molecular Biology Course
Authors: McKee Edward E.
Published: June 2021

Context

Team‐Based Learning is a well‐characterized and highly interactive pedagogy that has been used for some time in a variety of disciplines. However, its use in medical education and in Medical Biochemistry is more recent. The popularity of Team‐ Based learning appears to be growing as medical education strives to become more interactive, particularly because TBL is an interactive teaching strategy that can be delivered by a single faculty member. I have made extensive use of TBL for the past 5 years in my course in Medical Biochemistry and Molecular Biology.

Objectives

The poster will describe the basics of TBL and how TBL has been used in my course to integrate and apply content. The added value of TBL in a competency based curriculum is also described. TBL is a basis for students to practice, refine, and assess their peers on a variety of competencies required for team interaction, including: communication, life‐long learning, self awareness, ethical and moral reasoning, problem solving, and professionalism.

Conclusion

The TBL sessions in this course were highly valued by the students and provided a highly interactive environment for learning content and perfecting team skills.
Cadmium Inhibition of the Sodium‐Glucose Transporter
Authors: Lozier Emilie , Murali Ananya , Murali Anjana , Murali Lalitha , Kothinti Rajendra K. , Niloofar Tabatabai
Published: June 2021

Cadmium (Cd2+) is a toxic metal found in the environment, a product of industrial pollution, and can enter the body through inhalation. Chronic exposure to Cd2+ causes kidney failure characterized by Fanconi‐like Syndrome, in which an array of sodium‐dependent nutrient transport is inhibited. Cd2+ causes a concentration‐dependent reduction in sodium‐glucose co‐transport that is correlated to the sodium‐glucose transporter (SGLT1) gene expression in kidney proximal tubule epithelial cells. SGLT1 is regulated by the zinc finger transcription factor, Sp1. In the presence of Cd2+, the zinc ion is displaced by a Cd2+ ion, resulting in the loss of transcriptional regulation of SGLT1. The structure of human SGLT1 is not known; as such the Shorewood SMART Team (Students Modeling A Research Topic) modeled a bacterial homolog of SGLT1 using 3D printing technology. The SGLT1 protein transmembrane helices (TM2E, TM3, TM7E, TM8, and TM11) and amino acids on these helices vital for Na+ dependent glucose co‐transport are conserved. SGLT1 undergoes a conformational change enabling Na+ dependent glucose transport across the apical membrane of proximal tubule of kidney. In order to understand the glucose transport in the face of Cd2+ and to understand the underlying molecular mechanism responsible for SGLT1 glucose transport, it is essential to know the protein structure of SGLT1. Supported by a grant from NIH‐SEPA and NIH‐CTSA
Assessments of undergraduate student writing in the sciences
Authors: Smith Sally Kathryn Sommers , Griffin Harry , Lavalli Kari
Published: June 2021

Undergraduate science educators at the university level have noted the difficulty many beginning students have in generating satisfactory scientific writing. This study was designed to assess undergraduate scientific writing and student attitudes about writing in the sciences. We examined and quantified specific areas of scientific with which students had difficulty as well as the results of opportunities to revise submitted work. Areas of difficulty included failing to clearly illustrate the purpose of the study as well as failing to formulate, discuss and link a stated hypothesis to the results of a study. Statistical methods were applied to determine whether significant improvements had been made in revised papers. We also looked at the pre‐existing attitudes of students towards the practice of writing in the sciences and what effects these attitudes may have on their ability to communicate well in a science writing assignment. The results of this study provide educators with some insight necessary to craft writing assignments that challenge students as well as support their scientific writing efforts. Because scientific writing is the vehicle for the dissemination of new and important discoveries, advancing students’ abilities to generate and comprehend scientific writing will benefit students as they move into their major areas of study.
Modeling of M2‐PYK to show Possible ″Drugable Sights″ in Cancer Cells
Authors: Wuelzer Jennifer , Schultz Christine , Patel Nehal , Fiscus Allie , Bordewick Bridgette , Dix Randy , Fenton Aron W.
Published: June 2021

An important discovery in cancer biology is that the expression of pyruvate kinase M2 (M2‐PYK) as the basis of cancer specific metabolism. M2‐PYK is a protein that catalyzes the last step of glycolysis by transferring the phosphate group from the phosphoenolpyruvate (PEP) to adenosine diphosphate (ADP) resulting in pyruvate and adenosine triphosphate (ATP). During the Warburg effect, tumors show an increased rate of glycolysis, but instead of pyruvate moving to the Krebs cycle, it enters the lactic acid fermentation pathway. The isozyme, M2‐PYK, dissociates from the active tetramer into two inactive dimers. The Olathe North SMART Team (Students Modeling A Research Topic) modeled the M2‐PYK using 3D printing technology to show possible drug binding sites designed to stabilize the tetramer against dissociation. Our mentor provided tools to locate potential “drugable pockets” on the surface of the protein that are created by residues from the two neighboring subunits across the interface. The criteria used to identify a drugable pocket were: 1) the presence of multiple side‐chains that could be ionized to carry a charge that come from each subunit to make up the interface; and 2) a surface that reflects some level of penetration into the protein, i.e. a fissure in the protein's surface when every atom in the protein is rendered by its Van der Waals radius. Supported by a grant from NIH‐SEPA.
Regulation of the suppressor of cytokine signaling 3 (SOCS3) by zinc
Authors: Liuzzi Juan , Tracey Ashley
Published: June 2021

Zinc is an essential trace element that is known to have anti‐inflammatory activity; however, the mechanisms by which zinc inhibits inflammation are not completely understood. Promoter analysis of the family of suppressors of cytokine signaling (SOCS) revealed that the SOCS3 gene contains four binding sites for the metal regulatory transcription factor 1 (MTF1) located within 1800 bp upstream of the transcription start site. Hence, we hypothesized that zinc regulates the expression of SOCS3 through the activation of the transcription factor MTF1. A series of experiments were carried out with human hepatoma cells (HepG2) to examine the role of zinc and MTF1 in the regulation of SOCS3. We evaluated the effect of zinc excess on the expression of SOCS3 mRNA in cells treated with or without actinomycin D. In addition, we tested the role of MTF1 in the regulation of SOCS3 expression using chromatin immunoprecipitation (ChIP) assay and siRNA. Results indicate that SOCS3 expression is induced by zinc at the transcriptional level. Furthermore, ChIP assay revealed that zinc treatment increases MTF1 binding to the promoter of SOCS3 gene. Lastly, MTF1 knockdown caused a significant reduction of the basal expression of SOCS3. The results suggest that zinc regulates SOCS3 in a MTF1 dependent manner; however, more studies are needed to determine the possible involvement of SOCS3 in the anti‐inflammatory activity of zinc.
Semi‐quantitative analysis of milk proteomes reveals new evolving activities in carbohydrate metabolism in breastfeeding women
Authors: Gao Xinliu , Zhang Qiang , McMahon Robert J , Woo Jessica G , Davidson Barbara S , Morrow Ardythe L
Published: June 2021

Human milk proteins are thought to support a number of fundamental biological processes in the neonate. Among these processes may be carbohydrate metabolism, as lactose is abundant in the diet of the breastfed infant. Here we present semi‐quantitative analysis of whey proteins of human colostrum (first week, n = 3) and corresponding mature milk (3 mo.) by shotgun proteomics. Extended Gene Ontology of the identified proteins reveals that carbohydrate metabolism gene products are well represented in human milk. 80 proteins related to carbohydrate metabolism were identified, 24 of which were found to be significantly higher in mature milk. The diverse array of human milk proteins related to carbohydrate metabolism and their higher abundance in mature milk may suggest a role for human milk in adapting to increasing carbohydrate content during breastfeeding. The research is supported by Mead Johnson Nutrition and NIH HD 13021.
Effect of long‐term high‐fat diet feeding on energy metabolic substrates utilization in resting rats
Authors: Hwang Hyejung , Kim Jisu , Suh Heajung , Lee Seongno , Lim Kiwon
Published: June 2021

This study investigated the effect of long‐term high‐fat diet feeding on energy metabolic substrates utilization in resting rats and to revalue source fat energy efficiency using high‐fat diet feeding for measure energy expenditure and body fat accumulation in resting rats. Which were divided into control (CON) group and high‐fat diet (HF) group. Rats had fed high fat diet (w/w 40%) adlibitum for 5 weeks. Energy expenditure was measured using animal energy metabolism chamber after 4 weeks feeding. Final body weight did not changed between CON and HF groups, but caloric intake was significantly higher in HF group than in CON group (p<0.05). There was no difference between CON and HF groups in oxygen uptake, however carbon dioxide production showed significant higher in HF group. Also, respiratory exchange ratio was higher in HF group. Carbohydrate oxidation was lower in HF group than in CON group, but fat oxidation in HF group was significantly greater. These results mean that energy substrates oxidation at rest is affected by diet composition, especially dietary fat content. Abdominal fat fad weights were significantly higher by 33% in HF group than in CON group even though the calorie intake in HF group was higher by 6%. These results suggested that the dietary fat calorie value might be higher Atwater value of 9 kcal/g, and mean that dietary fat calorie value could be reconsidered in obese or weight control athletes.
Influence of short‐term nutritional overload on inflammationrelated gene expression profile in adipose tissue of Wistar rats
Authors: Francisqueti Fabiane Valentini , Pierine Damiana Tortolero , Nascimento Andre Ferreira , de Azevedo Melo Luvizotto Renata , dos Anjos Ferreira Ana Lucia , Cicogna Antonio Carlos , Correa Camila Renata
Published: June 2021

High consumption of fat and sugar is linked to obesity and induces adipocyte to secrete proinflammatory cytokines. However, it is unclear when this process occurs. The aim was to determine the effect of short‐term of high‐saturated fat diet plus sugar on obesity and inflammatory cytokines gene expression in adipose tissue. Male Wistar rats (body weigth =348±34g) received standard chow (C, 25% protein, 58% carbohydrate and 5% fat 17%, with 17% saturated and 82% unsaturated) or high‐fat diet (H, 21% protein, 45% carbohydrate, 29% fat with 50% fat saturated and 50% unsaturated fat) plus sugar in the water (30%). Obesity was characterized by adiposity index (%). Was assessed gene expression of inflammation‐related gene (IL‐6, TNF‐α and MCP‐ 1). H rats showed an adiposity index higher than C (C= 4,6 ±0,9 vs H= 7,7±1,5%, p< 0,001). There was no diference between groups for inflammation‐related gene in adipose tissue. The short‐term of nutritional overload leads to obesity, but was not enough to alter the inflammation‐related gene expression profile in epididymal adipose tissue.
Using Clinically Relevant Approaches in Teaching of Anatomical Science Courses to Medical Students
Authors: Cui Dongmei , Haines Duane E , Lynch James C , Yang Gongchao , Conway Marianne L
Published: June 2021

In recent years, medical schools have recognized the importance of integrating clinical information into the teaching of basic sciences. Many efforts have been made by educators to engage clinical relevance in basic science courses to bridge the gap between the basic science and clinical education. At the University of Mississippi Medical Center, we have integrated clinical information into our anatomical science courses using various approaches that have directly or indirectly transformed the courses and have provided a clinical context. These methods include (1) using clinical terminology during lectures and laboratory exercises; (2) using clinical cases and examples to make anatomic points; (3) using clinical vignettes in discussion sessions and in examinations; (4) inviting clinicians to participate in teaching our basic courses; (5) using textbooks that balance basic science and clinical knowledge; (6) using MRI, CT in teaching anatomy, preparing M1 students for future clinical years. In summary, These approaches pave the road for medical students to develop critical thinking skills and better prepare them to transfer the basic anatomic knowledge in their clinical practice.
Contribution of Unilever Reformulation to Changes in Nutrient Intakes: Modeling of Sodium & SAFA
Authors: Roodenburg Annet Johanna Christina , Dotsch-Klerk Mariska
Published: June 2021

Based on the WHO Strategy for Diet Physical Activity and Health (2004), the food industry has been reformulating products into a healthy direction. In 2005, Unilever started as one of the first global food companies with an active programme aimed at food reformulation; now continued under the Unilever Sustainable Living Plan.

To estimate the effect of reformulation by Unilever of specific foods in the period 2005–2007, the Monte Carlo Risk Assessment model was used. Inputs for this model were food intakes from surveys and nutrient composition of foods (NEVO) as well as data from Unilever (including market shares).

Results

Sodium intake from soups was reduced 7%–19%. In absolute numbers this was 10–20 mg sodium: representing <1% of total daily sodium consumption (> 2400 mg/d) for the total population.

SAFA intake from spreads and oils was reduced 12–33%. This was 0.6–1.4 g/d per day: 3–4% of total daily SAFA consumption (32g/d) for the total population.

The analyses show changes in nutrient intakes due to reformulations within product groups. The public health relevance depends on the relative importance of these product groups in the total food consumption pattern of a population.
Bioaccesibility of Carotenoids from Maize Flour with Varying Levels of Resistant Starch Type 2 and 3
Authors: Surles Rebecca Lynn , Tanumihardjo Sherry , Palacios Natalia
Published: June 2021

Background

It has been proposed that resistant starch may decrease the bioavailability of provitamin A carotenoids from some foods possibly by interfering with micelle formation in the gut. Research is needed to determine if the type of resistant starch (RS) and processing methods of food like maize influence bioavailability of carotenoids. To do this it is first necessary to assess interactions between provitamin A carotenoids and resistant starch content.

Object

Three Stage In Vitro Digestion Method will be used to assess micellarization efficiency of carotenoids from maize flour with various concentrations of resistant starch to determine bioaccesibility of provitamin A.

Study design

Type of RS in the maize flour depends on the processing method. Uncooked flour will have higher proportions of resistant starch 2 (RS2) whereas cooked and cooled maize flour will be higher in the retrograded starch or resistant starch 3 (RS3). Starch in cooked maize will be gelatinized increasing the availability of starch (AS). Total RS will be analyzed with a Megazyme kit and carotenoids will be quantified before and after in vitro digestion in maize flour with various carotenoid and RS concentrations. Total starch and oil will be quantified to assess bioaccesibility interactions. Results from this study will provide necessary research towards understanding provitamin A bioaccesibility in maize. Supported by Harvest Plus.

*All the following procedures will be conducted in the cooked and raw samples

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*All the following procedures will be conducted in the cooked and raw samples
The MTHFR C677T gene polymorphism controversy. A protective effect for renal function in patients without hyperomocysteinemia under adequate alimentary regimen
Authors: Trovato Guglielmo M , Catalano Daniela , Ragusa Angela , Trovato Francesca M. , Pirri Clara , Di Nora Concetta , Buccheri Maria Antonietta , Tonzuso Antonia , Martines G. Fabio
Published: June 2021

The association with atherosclerosis, renal failure and Cardio‐Vascular events of MTHFR gene C677T polymorphism is still controversial.

Aim

to investigate relationship of MTHFR 677C>T polymorphisms with Renal Function in non‐diabetic subjects without hyperhomocysteinemia. We studied 178 subject: 78 MTHFR C677CC (no polimorphism), 52 HFR C677CT and 48 MTHFR C677TT (with polymorphism) patients (age 50.93±14.85 yrs, BMI 27.95±5.98 kg/m2), all with adequate adherence to Mediterranean Diet Italian Guidelines and normal serum homocysteine. Glomerular Filtration Rate (eGFR), Insulin Resistance (HOMA), Parathyroid Hormone, UltraSound Doppler Renal Resistive Index of renal artery and Echocardiography Left Ventricular Mass/m2 were assessed.

Results

MTHFR 677C>T polymorphism has higher eGFR (73.8±27.99 vs. 58.64±29.95; p<0.001) and lower Renal Failure risk (OR 0.684; CI 95% 0.313–1.494) in comparison with controls. By multivariate analysis (MLR), MTHFR polymorphism favorable effect is independent from the negative effects of insulin resistance, hyper‐parathyroidism and obesity. Conclusiono. MTHFR 677C>T gene polymorphism is associated with better renal function in well‐nourished patients, independently from other variables and homocysteine levels.
Dietary consumption of eggs does not cause hypercholesterolemia and may not be indicated in the development of cardiovascular disease
Authors: Johnson Sarah Brooke Ardanuy , Chai Sheau Ching , Hooshmand Shirin , Saadat Raz , Arjmandi Bahram H.
Published: June 2021

Although hypercholesterolemia (HC) is regarded as a major risk factor for cardiovascular disease (CVD), approximately 50% of Americans die from CVD without ever having HC. People often limit their egg consumption to reduce their cholesterol intake, in turn also depriving themselves of other nutrients known to have positive effects on overall CVD health. The objective of the present study was to examine the degree to which CVD risk factors were influenced by egg consumption. In a cross‐sectional study, 146 healthy, postmenopausal women were divided into 1st, 2nd, 3rd, and 4th quartiles, respectively (<1 egg/week, 1 egg/week, 2–6 eggs/week, and >7 eggs/week). Overnight fasting measurements included lipid profiles, glucose, and inflammatory markers. Pearson correlation coefficients were used to determine the relationship between variables of interest. The 2nd quartile was associated with increased HDL‐C levels (82±3 mg/dL) and reduced TG levels (78±5 mg/dL). The 4th quartile was associated with reduced TG levels (93±20 mg/dL), and all other variables, including TC, were not correlated. These findings suggest that egg dietary cholesterol does not cause HC and may not be indicated in the development of CVD. Source of funding: USDA/NRI.
Altered fasting human plasma metabolite profile associated with short‐term blackberry feeding
Authors: Nicastro Holly , Novotny Janet A , Clevidence Beverly A , Charron Craig S , Dawson Harry D , Milner John A
Published: June 2021

Healthy adults (n=51) over the age of 40 consumed a wash‐in diet for 3 d followed by an intervention diet with 300 g blackberries or an isocaloric gelatin control at breakfast for 6 d. Metabolite profiling was conducted by Metabolon using GC/MS and LC/MS/MS platforms using fasting plasma samples from d 9. Following log transformation and imputation with minimum observed values, Welch's two‐sample t‐test was used to identify biochemicals that differed significantly between experimental groups. Blackberry intake compared to control revealed metabolite changes that reflect likely changes in gut metabolism (phenol sulfate), TCA cycle energetics (α‐ketoglutarate), the pentose phosphate pathway (threitol), and metabolism of benzoate (phenol sulfate), isoleucine (3‐hydroxy‐2‐ethylpropionate), medium‐chain fatty acids (undecanoate), collagen (trans‐4‐hydroxyproline), choline (betaine), and xanthine (3‐methylxanthine) and decreased fucose. Methyl‐beta‐glucopyranoside, a likely anthocyanidin metabolite, and one unknown compound were identified as potential markers of compliance for blackberry intake. Levels of these markers, however, did not correlate with levels of other altered metabolites. Support: NIH/USDA
Hass Avocado Improves Postprandial Vascular Reactivity When Consumed in Combination with Cooked Hamburger Meat
Authors: Li Zhaoping , Zhang Yanjun , Henning Susanne M. , Wong Angela , Jones Alexis , Zerlin Alona , Thames Gail , Bowerman Susan , Heber David
Published: June 2021

The Hass Avocado contains monounsaturated fats (oleic acid) and antioxidants (carotenoids, tocopherol). Cooked hamburger meat contains lipid peroxidation products formed during cooking. After ingestion, the stomach functions as a bioreactor, generating lipid peroxides and this process can be inhibited when antioxidants are ingested together with the meat. The present study was conducted in eleven healthy subjects on two separate occasions consuming either a 250 gram hamburger patty alone (ca. 436 Cal and 25 g fat) or the hamburger together with 68 grams of avocado flesh (an additional 114 Cal and 11 g of fat for a total of 550 Cal and 36 g fat), a common culinary combination, to assess effects on vascular health. Following a standard peripheral arterial tonometry (PAT) method (Itamar) to calculate PAT index there was a significant decrease 2 hours following hamburger ingestion alone (2.19 ± 0.36 vs. 1.56 + 0.21, p=0.0007) which was prevented when avocado was ingested with the burger (2.17 ± 0.57 vs. 2.08 ± 0.51, NS (p= 0.68). Monocytes were isolated from postprandial blood samples and I kappa‐B alpha (IKBa) protein measured to assess effects on activation of inflammation. At 3 hours, there was a significant preservation of IkBa (131 % vs. 58 %, p=0.03) consistent with inhibition of inflammation via the NF‐kappa B pathway. Triglycerides, MDA, IL‐6, and TNF‐alpha increased postprandially but were not further increased when avocado was ingested with the burger compared to burger alone, despite the added fat and calories from the avocado. These observations are consistent with beneficial antioxidant and vascular health effects of ingesting added Hass avocado with a hamburger patty.

Study Supported by The Hass Avocado Board
A high fat/high fructose (HF) diet does not exacerbate the metabolic or cardiac functional consequences induced by diabetes in C57Bl6J mice
Authors: Lachance Dominic , Vaillant Fanny , Shi Yanfen , Daneault Caroline , Frayne-Robillard Isabelle , Deschepper Christian , Carpentier Andre , Tardif Jean-Claude , Rosiers Christine Des
Published: June 2021

Background & Objectives

High fat or high sugar diets are associated with increased risk for coronary heart disease but have not been found to exacerbate cardiac dysfunction in rodent models of heart failure. We thus tested the effect of HF diet in a mouse model of diabetes and its associated cardiac dysfunction.

Methods

Five‐week‐old C57Bl6J male mice received either standard (Std) or HF diet for 2 weeks, then received multiple injections of either low doses (40 mg/kg) of streptozotocin (STZ) or vehicle (Veh).

Results

Ten weeks after STZ injections (and compared to Std‐Veh mice), both STZ groups displayed similar (i) fasting plasma glucose and insulin levels, which were respectively increased and decreased (p<0.001) and (ii) cardiac systolic dysfunction, as evidenced by a 8% lower ejection fraction (p<0.05) at echocardiography. However, the increase in plasma levels of ketone bodies, glycated haemoglobin and branched chain amino acids was smaller (p<0.05) in HF‐STZ mice than in Std‐STZ at both 6 and 12 weeks. Likewise, HF‐STZ mice maintained (in contrast to Std‐STZ mice) body weight and epidydymal fat mass as well as Std‐Veh.

Conclusion

Not only did HF not exacerbate diabetes‐induced cardiac dysfunction, but it even attenuated the systemic metabolic alterations induced by STZ, which may in the long term delay the development of heart failure. (Supported by CIHR & HSFC)
Grains, nuts and fruits dietary pattern is associated with a reduced likelihood of having metabolic syndrome
Authors: Kang Minji , Park Young-Hee , Baik Hyun Wook , Paik Hee Young , Joung Hyojee
Published: June 2021

The purpose of this study was to explore the relationship between dietary patterns and metabolic syndrome (MetS) and its risk components among Korean adults at risk of metabolic diseases. This cross‐sectional study included 280 subjects (152 men, 128 women) between 30 and 59 years of age recruited by the Bundang Jesaeng Hospital, Korea. MetS was defined based on Adult Treatment Panel III. Dietary data were collected using 3‐day food records. The percentage of energy intakes from 23 food groups was calculated, and a factor analysis was applied to identify dietary patterns. Four major dietary patterns were derived; “Grains, nuts and fruits (9.1%)”, “Bread, dairy and cereal, snacks (9.1%)”, “Meat and alcohol (7.1%)” and “Sweets, seasonings and oils (7.1%)”. In “Grains, nuts and fruits” pattern, participants in the highest quartile had lower odds ratios for MetS (OR: 0.43, 95% CI: 0.20–0.95) and high fasting glucose (OR: 0.42, 95% CI: 0.19–0.93) than those in the lowest quartile. Our study suggests that the “Grains, nuts and fruits” pattern has beneficial effects on the risk of MetS in Korean adults. This work was supported by the grant from the Cooperative Research Program for Agriculture Science & Technology Development (no. PJ007211), Rural Development Administration, Republic of Korea.
The apoptotic effect of allicin in MCF‐7 human breast cancer cells: role for ATF3
Authors: Park Bongkyun , Kim Kyoungho , Rhee Dong-kwon , Pyo Suhkneung
Published: June 2021

Allicin induces apoptosis in various carcinoma cells and inhibits tumorigenesis. Activating transcription factor 3 (ATF3) is a stress‐inducible gene and known as a transcription factor to induce or inhibit target genes. However, the precise mechanism of allicin‐induced apoptosis is still unclear in human breast cancer cells. We investigated the effect of allicin on cell proliferation, apoptosis, and its pathway in MCF‐7 cells. MTT assay and western blotting showed that allicin decreased concentration‐dependently cell proliferation, induced apoptosis related proteins (Bax, Bid, AIF) and activated caspase‐3 and 8 in a concentration dependent manner, respectively. Our data also showed that allicin increased phosphorylation of ERK1/2, p38 and JNK in a concentration‐and time‐dependent manner and protein level of ATF3. In addition, MTT assay and western blotting showed that allicin treatment resulted in decreased cell proliferation and high level expression of apoptosis related proteins in ATF3‐overexpressed MCF‐7 cells, respectively, suggesting that ATF3 expression is involved in the regulation of allicin‐induced apoptosis. Taken together, the data suggest that allicin induces apoptosis through MAPKs and ATF3 signaling pathways in MCF‐7 cells.
The inhibitory effect of Ramalin on the expression of VCAM‐1 in human aortic smooth muscle cells
Authors: Park Bongkyun , Rhee Dong-kwon , Pyo Suhkneung
Published: June 2021

Atherosclerosis is a chronic inflammatory disorder of the arteries. Since chronic inflammation increases various adhesion molecules, interfering with the expression of adhesion molecules may be an important therapeutic target of inflammatory disorder. Ramalin has been known to have a variety of biological activities including anti‐inflammatory activity. However, other biological activities of ramalin and its molecular mechanism remain unknown. We examined the effect of ramalin on the expression of the vascular cell adhesion molecule (VCAM‐1) and peptidylarginine deiminase IV (PADI4) in TNF‐α‐induced human aortic smooth muscle cells (HASMCs). Cell adhesion assay and western blotting showed that ramalin dose–dependently inhibited TNF‐α‐induced adhesion of THP‐1 monocytic cells and expression of VCAM‐1 and PADI4. In addition, inhibition of PADI4 activity by siPADI4 suppressed TNF‐α‐induced VCAM‐1 and c‐Fos protein expression, suggesting that PADI4 is involved in VCAM‐1 expression. Our data also revealed that ramalin inhibited phosphorylation of MAPKs and the nuclear translocation of NK‐κB and AP‐1 induced by TNF‐α. Overall, these results provide ramalin inhibits TNF‐α‐induced VCAM‐1 expression through the suppression of PADI4, MAPKs, NF‐κB and AP‐1 signaling pathways and may be considered in therapeutic strategy to reduce the risk of atherosclerosis.