bluuebunny/crossref_metadata_2025 huggingface dataset:
Prohibitin Plays an Important Role in Adipocyte Differentiation
Authors: Ande Sudharsana Rao , Xu Zuyuan , Gu Yuanyuan , Mishra Suresh
Published: June 2021

Insulin is a potent hormone that induces differentiation of preadipocytes into adipocytes (adipogenesis). However, various target genes required for the differentiation and in the maintenance of adipocytes are remaining to be identified. Herein we demonstrate that a nuclear coded mitochondrial protein prohibitin (PHB) is a target gene for insulin and play a modulatory role in adipogenesis. Treatment of 3T3‐L1 fibroblasts with insulin or peroxisome proliferator‐activated receptor‐gamma agonist resulted in an upregulation of PHB in a dose‐ and time‐dependent manner. An analysis of PHB promoter sequence revealed the presence of putative insulin‐response elements and CCAAT/enhancer‐binding proteins transcription elements within ~1 kb upstream of translation initiation site. Functional relevance of these sites was determined using reporter gene assay. Surprisingly, PHB was also found to be regulated by leptin. Furthermore, overexpression of PHB in 3T3‐L1 fibroblasts was sufficient to induce adipogenesis. In summary, we have identified PHB as an important protein in adipogenesis. This work was supported by funds from NSERC and MHRC. SM is a recipient of MMSF Career Development Award and SRA is supported by MHRC postdoctoral fellowship.
Real‐time detection of ethanol effects on cAMP in subcellular compartments
Authors: Gupta Ratna , Qualls-Creekmore Emily , Yoshimura Masami
Published: June 2021

Our studies have shown that acute ethanol (EtOH) exposure enhances GPCR‐stimulated adenylyl cyclase (AC) activity. However, the dynamics and intracellular distribution of cAMP is not known. Epac1‐camps, a FRET based cAMP sensor, was used to study the effect of EtOH exposure in three subcellular compartments: cytoplasm [Cyt], nucleus [Nuc], and plasma membrane [PM]. Hela cells were transfected with AC7 or AC3 isoform, dopamine (DA) D1A receptor, and FRET sensor targeted to different compartments. Time‐lapse FRET was used to monitor the cAMP level. Stimulation of cells expressing AC7 with DA showed an increase in cAMP in all the subcellular compartments. In the presence of DA and EtOH, the increase in cAMP was higher in all three subcellular compartments. In Cyt and Nuc the cAMP level reached a higher plateau in the presence of DA and EtOH. However, in PM the cAMP increased to a higher level transiently and decreased to a plateau similar to the one observed with DA alone. The results suggest that the level of cAMP is controlled by phosphodiesterase in a subcellular compartment specific manner and that the level of cAMP at PM may be under tighter control than in other compartments. EtOH had negligible effect on cAMP in all three subcellular compartments of cells expressing AC3. Therefore, the EtOH effect on intracellular cAMP is AC isoform specific and varies in different cellular compartments. Supported by NIAAA‐AA017303.
Structural studies of the regulatory domain of bovine soluble guanylate cyclase
Authors: Garcin Elsa D , Rassouli-Taylor Leida
Published: June 2021

Soluble guanylate cyclase (sGC) converts GTP to cGMP to promote vasodilation. The β subunit contains a heme nitric oxide/oxygen‐binding (HNOX) regulatory domain, an HNOX‐associated (HNOXA) and coiled‐coil dimerization domain, and a catalytic guanylate cyclase domain. Nitric oxide (NO) binds to the heme and allosterically stimulates the cyclase activity. We aim to determine the mechanism of NO‐induced activation of sGC. Ultimately, our results will facilitate the development of novel sGC activators to treat cardiovascular diseases.

We focused on two truncated constructs encompassing either the HNOX domain or both HNOX‐HNOXA domains. We used site‐directed mutagenesis to generate proteins that would be highly expressed and soluble in E. coli. From the 18 constructs generated, only 5 were expressed and soluble. Our preliminary results suggest that the HNOXA domain plays a role in proper folding of the HNOX domain and/or stability of the heme, in contrast to what was shown for bacterial HNOX homologs. Crystals of HNOX‐sGC diffracted poorly at the synchrotron, but to our knowledge, these are the first mammalian HNOX crystals. We are now working to improve the diffraction quality of these crystals.

Funding: American Heart Association Scientist Development Grant and UMBC DRIF.
Whole‐genome expression patterns in LTP knockdown Arabidopsis plants
Authors: Homayoun Yasmin , Femrite Chase , Vellanoweth Robert , Gracey Andrew
Published: June 2021

Abstract

Our lab studies the aging process in plants, using the plant model Arabidopsis thaliana. Arabidopsis’ small size and rapid life cycle make it an attractive model to study plant senescence. Previous research in our laboratory suggests that as Arabidopsis undergoes the developmental transition from vegetative growth to flowering, ascorbate peroxidase (APX) activity that is responsible for scavenging H2
O2 is repressed and 13‐lipoxygenase (LOX) is activated. LOX catalyzes the first reaction in oxylipin‐signaling pathways, which generate a multitude of lipid signaling molecules. We specifically focus on the possibility that lipid transfer proteins may carry signals during the aging process of the plant. Lipid transfer proteins are small, basic proteins present in the plant cell wall. Other data from our lab showed an up‐regulation of two lipid transfer proteins as the plant transitions from the vegetative to flowering stage. These data suggest that oxylipins may be involved as a signal between meristems and leaves during the vegetative to flowering transition in Arabidopsis. The purpose of this study is to compare gene expression patterns of Arabidopsis in wild type and LTP RNAi knockdown mutants, in meristems and flowers during different developmental stages and determine how gene expression patterns are altered in RNAi knockdowns. Our current RNAi knockdown mutants in Arabidopsis show uncontrolled growth producing an abundant number of flowers and seeds and live six times longer than an average wild type plant. Based on our knockdown phenotypes, we predict that downregulation of these genes may play a major role in the inhibition of meristem arrest.
Beneficial effects of Jamaican bitter yam biomaterials in hypercholesterolemic mice
Authors: Stennett Dewayne Kirkpatrick , Wheatley Andrew , Dilworth Lowell , McGrowder Donovan , Asemota Helen
Published: June 2021

The Jamaican bitter yam (Dioscorea polygonoides) is one of the wild yam varieties found in Jamaica and is a rich source of saponins, a wide variety of phytochemicals known to possess potent hypocholesterolemic properties with associated lipoprotein redistribution. These phytochemicals have also been shown to display hepatoprotective properties. The study therefore examined the hypolipidemic and hepatoprotective effects of bitter yam biomaterials (BYB) supplementation. Hypercholesterolemic mice were placed in four groups of eight mice each while normocholesterolemic mice were placed in two groups of eight mice each. They were then fed diets with or without BYB supplementation. Hypercholesterolemic controls had significantly higher (P<0.05) liver weights, serum total and LDL cholesterol, atherogenic index, lipid peroxidation in the livers and alkaline phosphatase and gamma glutaryl transferase activities when compared with normocholesterolemic controls. BYB supplementation led to significant reductions in all parameters except for the hepatomegaly experienced by the hypercholesterolemic controls. These results suggest that BYB supplementation may be beneficial in reversing the devastating effects of hypercholesterolemia and ameliorating liver damage.
Expression and Purification of Sunflower Thiolase Mutants
Authors: Dyer James H.
Published: June 2021

Sunflower (Helianthus annus L) cotyledons have two thiolase activities that have been identified previously. Thiolase II (acetoacetyl CoA thiolase) has specificity for the four‐carbon acetoacetyl CoA, resulting in the production of two Acetyl CoA molecules. Thiolase I (oxoacyl CoA thiolase) is active towards short, medium and long chain acyl CoAs. The purpose of this research was to optimize the expression and purification of mutant forms of these two thiolases from an E. coli bacterial expression system. Mutants generated by site‐directed mutagenesis were cloned into the pET151 expression vector, and constructs were transformed into BL‐21 E. coli that were induced for production of the thiolases with IPTG (Isopropyl β‐D‐1‐thiogalactopyranoside). The soluble thiolases were purified and were used to characterize their enzymatic properties.
Simultaneous Detoxification and Allergens Inactivation of Castor Bean Meal by Calcium Compounds Treatment and Solid‐state Fermentation
Authors: Fernandes Keysson Vieira , Deus-de-Oliveira Natália , Godoy Mateus Gomes , Nascimento Viviane Veiga , Melo Edésio José Tenório , Freire Denise Maria Guimarães , Machado Olga Lima Tavares
Published: June 2021

Ricinus communis has a great economical importance due to the extracted oil of its seeds that is used for the biodiesel synthesis. After oil extraction, a castor meal containing high protein level is obtained, however, it can not be used for animal consumption due to toxic (ricin) and allergenic (2S albumin) proteins. Two processes for castor meal detoxification and allergens inactivation were proposed. In addition, for detection of ricin, a biological test was introduced. In this test, Vero cells were treated with ricin and cell death was measured by cell counting and by measuring LDH activity. The minimum amount of ricin needed to kill 1.6 × 105 cells was 10 ng/mL. As a cake detoxification treatments, solid‐state fermentation (SSF) and treatment with calcium compounds were used. For SSF, Aspergillus niger was grown on castor mea, and this meal was analysed after 24, 48, 72 and 96. The ricin was eliminated after 24 hours of fermentation and after treatments with 4% and 8% of CaCO3, Ca(OH)2 and CaO. During SSF, lipases was also produced adding value to castor meal. By the other hand, after calcium treatment, the allergenic properties of castor cake, was totally abolished and a safe by‐product was obtained.
Deciphering the mechanism of antibiotic resistance associated with a plant ABC transporter
Authors: Ayalew Mentewab , Hardy Raven , Carlington Brittani , Davis Keithara , Nichols India , Jacobsen Megan
Published: June 2021

ATP binding cassette (ABC) transporters are found in most organisms and typically function in the transport of substrates across membranes. Atwbc19 is an Arabidopsis ABC transporter protein that confers antibiotic resistance. Overexpression of Atwbc19 results in increased kanamycin resistance in plants, but the mechanism has not been established. Atwbc19 has an N‐terminal nucleotide binding domain and a transmembrane domain with six alpha helices and thus was assumed to function as a transporter. Four internal GFP fusion constructs were generated and introduced in Arabidopsis to examine the localization of Atwbc19. However, Atwbc19 could not be localized to a membrane compartment. Instead, GFP fluorescence was observed in the nucleus and cytoplasm, suggesting cleavage of the N‐terminal soluble portion of the protein and subsequent nuclear localization. To generate an alternative hypothesis, we initiated the analysis of several highly conserved regions found in the protein sequence of Atwbc19. A yeast two hybrid screen was carried out using either the N‐terminal region or the linker region as baits to identify proteins interacting with Atwbc19. Two proteins identified in the yeast two hybrid screen are a Eukaryotic Initiation Factor 3 subunit and Casein Kinase 2. Our results suggest that Atwbc19 may not function as a transporter but as a translation regulator.
Column purification of histidine‐tagged proteins from unclarified samples
Authors: Carlsson Marianne , Lundqvist Jon , Granér Therese , Andersson Lars
Published: June 2021

The histidine‐tag is widely used to facilitate purification of recombinant expressed proteins. For immobilized metal‐ion affinity chromatography (IMAC) the most used metal ion is Ni2+. Also Co2+ has been shown useful in obtaining high purity with good yields of histidine‐tagged proteins. Prepacked HiTrap™ TALON® crude column facilitates scalable, fast and easy protein purification of histidine‐tagged proteins. Unclarified samples can be loaded directly to the column reducing the total purification time which helps to ensure high concentrated pure and intact target proteins with minimal degradation. Two proteins (produced in E. coli and P. pastoris) were purified ending up in >90% purity and yields of 90%. An automated purification of a kinase was done by IMAC and gel filtration. SDS‐PAGE analysis showed that the obtained purity of the kinase was above 90% and the amount purified protein was 3 mg. Further, four purification cycles, without cleaning and Co2+‐recharging between the runs, show that the HiTrap TALON crude columns can be used several times without loosing performance. Purity and yield was unchanged
Tamoxifen association to methacrylate polymer as a promising drug delivery system
Authors: Barros Susan S , Silva Jaqueline R , Azevedo Ricardo B , Gomes Anderson J
Published: June 2021

This study aimed to investigate the effect of a established cancer drug such as tamoxifen (TMX) and its encapsulation into a matrix of copolymer of ethyl acrylate, methyl methacrylate and a low content of methacrylic acid ester with quaternary ammonium groups (EUDRAGIT® RL 100) associated with a fluorescence probe such as fluorescein. TMX is used in breast cancer therapy with some side effects such as stroke, and endometrial cancer. The new drug delivery system (TMX‐NP) was designed to protect the drug from environmental and to promote a sustained drug delivery profile of the drug and also be tracked by the emission of fluorescence probe. The size and entrapment efficiency were evaluated at different preparation protocols. TMX showed a maximum entrapment efficient of 23% in the proportion of 1,5g of polymer and association to the fluorescence probe using a emulsion method. The size measurements were compared with the empty nanoparticles and showed values of TMX‐NP = 166.4± 32 nm, and empty NP = 273± 50 nm. The zeta potential of particles was +65 and +38 mV for TMX‐NP and empty NP respectively. The new drug delivery system posses some features parameter that allow it to be tested in biological samples such as cancer cells. Supported by FAPDF, CAPES, CNPq, FINATEC and DPP‐UnB.
Development of Analytical Methods for Characterization of Fab Fragments
Authors: Harwick Larissa Coombs , Fishpaugh Jeffrey R. , Rupprecht Kevin R.
Published: June 2021

The constant need for increased sensitivity of antibody‐based assays has increased the need for high affinity molecules. Use of Fab antibody fragments can increase the specificity and sensitivity of immunoassays. We developed simple GPC, SDS‐PAGE and IEF methods that can be used to characterize Fab material produced by reduction of F(ab’)2 fragments. Specific reduction of F(ab’)2 to Fab was accomplished using cysteamine HCl. The resulting Fab fragments were analyzed with and without alkylation with iodoacetamide. Fab fragments were analyzed by gel permeation chromatography using a Waters HPLC system and multiple column types. SDS‐PAGE was performed using multiple gel types and running conditions. Scanning densitometry was performed using a Bio‐Rad GS800 and electrophoretic profiles were determined. The pI range and IEF profiles of the Fab samples were determined using a Phastsystem (GE Healthcare). The methods developed provide reproducible profiles for both relative reduced fragment content and identity. The methods were capable of detecting low percentage impurities. These methods provide quick analytical protocols that can be used to characterize Fab fragments for use in immunoassays.
Identification and characterization of novel factors regulating kinesin‐1 activation and mitochondria dynamics
Authors: Cho Kyoungin , Patil Hemangi , Ferreira Paulo
Published: June 2021

The Ran‐binding protein 2 (RanBP2) is a mosaic and pleiotropic protein, which associates in vitro and in vivo with the conventional microtubule‐based motor and kinesin‐1 isoforms, KIF5B/KIF5C. We showed previously that the kinesin‐binding domain (KBD) together with the flanking Ran‐binding domains 2 and 3 (RBD2, RBD3) of RanBP2 directly jump‐start and boost the ATPase motor activity of kinesin‐1, a motor implicated in mitochondria transport. The goal of this study is to define the roles of RBD2 and RBD3 of RanBP2 and Ran GTPase in kinesin‐1 activation in vitro and multiple biophysical parameters of mitochondria motility in live cells. We show that the activation of kinesin‐1 by RBD2‐KBDRBD3 is Ran GTPase independent; yet mutations in RBD2‐KBDRBD3 and abolishing its interaction with Ran GTPase suppress the Michaelis‐Menton and partially affect the allosteric activation phases of kinesin‐1. These effects lead to a ~20% reduction of kinesin‐1 activity (kcat). In live cells, KBD alone stimulates the motility, displacement, frequency of change in direction, motility persistence and anterograde velocity of mitochondria, whereas a mutant KBD construct has no effect. In contrast, RBD2‐KBDRBD3 of RanBP2, but not mutant constructs, suppress all these biophysical parameters of mitochondria motility. Further, there is stereochemical specificity between RBD2 and RBD3 in certain parameters of regulation of mitochondria motility. These studies uncover novel Ran GTPase‐independent functions of RBD2, RBD3 and KBD of RanBP2 in kinesin‐1 activation and regulation of mitochondria motility.

NIH support GM083165 & EY019492 and RPB
A novel mitochondrial ubiquitin ligase involved in the regulation of mitochondrial fusion
Authors: Benischke Anne-Sophie , Hemion Charles , Neutzner Melanie , Norris Kristi , Frank Stefan , Youle Richard , Karbowski Mariusz , Neutzner Albert
Published: June 2021

Mitochondrial protein quality control is one of the mechanisms that protect mitochondrial integrity. Recognition of terminally misfolded proteins is achieved through outer mitochondrial membrane‐associated degradation (OMMAD). Misfolded proteins are ubiquitinated, retrotranslocated by the AAA‐ATPase p97 and then degraded by the proteasome. OMMAD ubiquitin ligases such as MARCH5, Parkin and IBRDC2 are key proteins in the control of mitochondrial fission, mitophagy and apoptosis, respectively. We describe an ubiquitin‐ligase (RTM9) which is localized on the outer mitochondrial membrane. RTM9 consists of transmembrane domains and a RING finger domain, exposed to the cytosol. In an ubiquitination assay RTM9 shows E3 ubiquitin ligase activity. RTM9 is a novel regulator of mitochondrial fusion: while expression of wildtype RTM9 had no effect on mitochondrial morphology, expression of RTM9RING‐ resulted in the fragmentation of the mitochondrial network. In addition, RTM9 interacts with both mitofusins, Mfn1 and Mfn2, suggesting a role for RTM9 in the regulation of mitochondrial fusion.

Taken together, RTM9 is a novel mitochondrial ubiquitin ligase involved in the regulation of mitochondrial fusion potentially playing a critical role during mitochondrial quality control.
Acute and Chronic Exposure of Nickel Promotes Breast Cancer Cell Growth
Authors: Sabangan Jackielyn Perez , Louie Maggie
Published: June 2021

Increasing incidences of breast cancer have been a global public health concern, however the underlying cause remains unknown. Studies have suggested that exposure to elevated levels of heavy metals has been associated with deregulated cell growth and cell death. Heavy metals including nickel (Ni2+) are known to mimic the effects of estrogen and promote breast cancer cell growth, but its mechanism of action is unclear. To understand Ni2+ carcinogenicity, we examined the effects of Ni2+ on ER+ breast cancer cell (MCF‐7) growth. Our results suggest that acute and chronic Ni2+ exposure increases MCF‐7 cell growth. Further analysis at the molecular level indicate that the expression of several cell cycle genes were upregulated, whereas the expression of two cyclin‐dependent kinase inhibitors (CKI), p21 and p27 were significantly decreased in response to Ni2+ exposure. The ability of CKIs to induce cell cycle arrest is dependent upon their interaction with their respective cyclin/cyclin dependent kinase complex. Understanding the molecular mechanism of how nickel regulates the expression of CKIs, and the role of ERα in this process will provide insights to how Ni2+ contributes to breast cancer etiology.
A mutant of Leishmania donovani deficient in elongating mannosylphosphoryl transferase activity
Authors: Phillips Megan R. , Turco Salvatore J.
Published: June 2021

Leishmania parasites manipulate essential interactions with its sand fly vector and vertebrate host by expressing developmentally‐regulated forms of its surface lipophosphoglycan (LPG). LPG is a GPI‐anchored polysaccharide consisting of repeating 6Galβ1,4Manα1‐PO4 units. The number of repeat units varies depending on the stage of the life cycle: n~15 in procyclic and n~30 in metacyclic promastigotes; and n=0 in the intracellular amastigote forms.

Our work focuses on the family of mannosyl‐phosphate transferases involved in LPG formation. At least two mannosylphosphoryltransferases (MPTs) have been implicated in repeat unit assembly: an “initiation” specific MPT (iMPT) which uses the LPG core as acceptor and an “elongation” specific MPT (eMPT) which uses repeat units as acceptor. We previously identified a chemical mutant of L. donovani that we showed to have only one Gal‐Man‐P unit in its LPG by being deficient in eMPT activity. We are currently examining the mutant for its potential in elucidating further molecular information regarding the critical eMPT that is essential in determining the length of the LPG phosphoglycan.

The funding source for this research is NIH grant is RO1‐AI31078 awarded to Salvatore J. Turco.
Avocado oil increases cell viability and respiratory rate of yeast during oxidative stress induced by ferrous iron
Authors: de la Paz J. Lucio Fernández , de Jesús Martínez-Morales Pedro , Calderón-Cortés Elizabeth , Fernández-Quintero Maricela , del Carmen Montoya-Pérez Rocío , Clemente-Guerrero Mónica , Saavedra-Molina Alfredo , Cortés-Rojo Christian
Published: June 2021

Mitochondrial dysfunction due to oxidative damage it has been linked to several pathological conditions, which has lead to an intense search for molecules with antioxidant properties at mitochondrial level. Avocado oil (AO) is a natural product that constitutes a source of lipophilic antioxidants. To evaluate the possible protective effects of AO against cellular and mitochondrial oxidative damage, we tested this oil in yeast cells subjected to oxidative stress induced by Fe2+. The growth of cell cultures in the presence of a quantity of AO equivalent to 10mM oleic acid increased cell viability in 36% and 11% after the exposure to 25 and 200 μM Fe2+, respectively. Avocado oil induced an increase of approximately 75% in both state 4 and uncoupled respiration at concentrations of Fe2+ up to 100 μM, while at 200 μM, respiration in uncoupled state was detected only in AO cells. No lipoperoxidation was detected at any condition studied. These results suggest that AO increase resistance of cells to oxidative stress by a lipoperoxidation‐independent mechanism by possibly increasing the respiratory capacity of the cell. This work was supported by CONACYT (130638 to CCR) and PROMEP (PTC‐266 to CCR) grants.
Identification of mechanism of action of anti‐HIV properties of compounds present in Neem (Azadirachta indica) extracts
Authors: Arora Sonia , Vanniyasingam Jananiga
Published: June 2021

Neem (Azadirachta indica) is a tree in the mahogany family Meliaceae. It is one of two species in the genus Azadirachta, and is native to Indian subcontinent. In India, Neem is widely regarded as “village pharmacy” because almost each part of the plant has been used for several different medicinal purposes. Neem extract prepared from its bark, leaf and seed has been shown to possess anthelmintic, antifungal, antibacterial, and anti‐malarial properties. Recently, in an in vitro study crude extract prepared from Neem leaves showed anti‐HIV‐1 activity. However the exact mechanism of action of this remains unknown. Hence the major goal of this project is to identify the molecular target of anti‐HIV1 action of Neem compounds. We hypothesize that compounds found in Neem may interact with various proteins essential for either entry or replicative life cycle of the HIV‐1 virus. Using structural bioinformatics approaches a small database of all the compounds known to present in crude extracts of Azadirachta indica was created. In silico docking was next used to study binding affinity of these compounds with several different molecular target proteins important in HIV‐1 life cycle viz. gp41 entry protein, reverse transcriptase, integrase and protease. Predicted binding affinity and interaction pattern emerged from in silico studies was further used to match compounds with their most likely protein target. Interestingly, our deductive analyses showed that most compounds present in Neem extract interact very strongly with HIV‐1 protease, hence making it plausible molecular target. In vitro assays are currently underway to further verify our in silico data. This research is supported by funds from Kean University and Ronald E. McNair Scholars Program.
CASP‐8 D302H gene polymorphism and susceptibility to primary brain tumors
Authors: Pence Sadrettin , Cacina Canan , Turan Saime , Genc Feyza , Kafadar Ali , Kaynar Mehmet Yasar , Eraltan Ilhan Yaylim
Published: June 2021

Caspase‐8(CASP‐8)is an initiator CASP in the cell death receptor mediated apoptotic pathway, and plays a crucial role in carcinogenesis. Our aim was to investigate the association of CASP‐8 D302H gene polymorphism with primary brain tumors susceptibility. CASP‐8 D302H genotypes were determined by using a polymerase chain reaction‐restriction fragment lenght polymorphism analysis. A total of 91 patients with primary brain tumors including and 114 control individuals were enrolled in the current study. There was significant difference in the distribution of CASP‐8 genotypes between patients with primary brain tumors and controls. We found that CASP‐8 D302H GG/CC homozygous genotypes have a significant 1.28‐fold increased risk in developing gliomas as compared with controls. These results suggest that D302H polymorphism in CASP‐8 gene may be associated with increased risk of primary brain tumors.
Monkeys, beach balls, and Twinkies®: using analogies to teach enzyme kinetics
Authors: First Eric Allen
Published: June 2021

Traditionally, enzyme kinetics is taught by emphasizing the mathematics behind the kinetic equations. For example, students are shown how to derive and transform the Michaelis‐Menten equation and calculate Km, kcat, and Ki values from various plots. Many, if not most, students respond by memorizing formulas and using a “plug and chug” approach to solve kinetic problems. This is unfortunate, as the students never gain an appreciation of how the kinetic analysis relates to the catalytic mechanism. An alternative to the above method is to use analogies to emphasize the physical picture behind the mathematics. This latter method gets students out of their comfort zone by forcing them to connect the physical picture to the reaction mechanism and, ultimately, the mathematical analysis. Students gain an appreciation of where the kinetic equations come from and develop a more flexible approach to problem solving. By encouraging interaction between the students and teacher, the analogies make teaching more effective and learning more enjoyable. Monkeys eating peanuts (“peanutases”) and students opening beach ball valves (“beachballases”) are especially useful analogies for teaching concepts in enzyme kinetics, including the meaning of Km and kcat, saturation kinetics, and enzyme inhibition. As for Twinkies®, they are both a delicious snack and a great way to explain the difference between thermodynamics and kinetics.
Promoting and Supporting Innovation Through Collaborative Research
Authors: Gentile Jim
Published: June 2021

Many of the major scientific challenges of the 21st century are likely to require collaborative and interdisciplinary teams. Creating good “team players” who can work synergistically to undertake high‐risk, high‐reward research should be a priority, and demonstrating success in collaborations should be encouraged by private and federal granting agencies, and more highly rewarded by universities in tenure and promotion decisions. Given the challenges that our nation faces, tenure and promotion decisions should no longer be based merely on the number of papers published or overhead dollars generated. Journal editorial boards and foundation peer‐review panels are essential to the highest‐quality science, but they should not be viewed as surrogate members of tenure and promotion committees. Rewarding careers paths should be clearly visible to the nation's newly minted science Ph.D.s, postdoctoral researchers and “tenure track” professors. Allowing these early‐career scientists more expansive opportunities during their most creative years would make the United States, and the academic communities in which they serve, an increasingly more vibrant and creative environments. The heart and soul of American science and enterprise is the willingness to take risks, to learn from failure and to repeat the process until success is achieved. Our early‐career scientists should have the same opportunity.
Getting the high schools involved in scientific research! The New York City Partnership to Advance Science (NYCPAS); Collaborative research projects between NYC high schools and a liberal arts college: The pilot year
Authors: Aguanno Ann
Published: June 2021

Marymount Manhattan College (MMC), a liberal arts college in New York City, has a small but growing Division of Sciences. To meet the need for more graduates in STEM fields, the division received funding from the U.S. Dept. of Education to establish the New York City Partnership to Advance Science (NYCPAS). This program fosters research collaborations between NYC high schools and the MMC Science Division by funding small research projects devised by high school students and their science teachers, under the guidance of MMC science faculty and undergraduates. Through these collaborative projects the program aims to enhance science education at the high school level, to establish long term connections between local high schools and MMC science faculty, and to recruit potential MMC science majors. Over two hundred high schools were solicited for proposals in fall 2010. Science faculty were available to help each high school team devise a research proposal. Thirteen proposals were funded and matched with MMC science faculty and upper level undergraduate mentors. High school teams implemented their projects during the spring of 2011 and had access to MMC facilities and instrumentation if needed. Research findings were then presented in poster format in a competitive symposium in June 2011 on the MMC campus. Assessment of this pilot year reveals that this program is a model for engaging and recruiting K‐12 students in science.
Characterization of a compensatory mutant of Leishmania major that lacks ether lipids but exhibits normal growth and G418 resistance
Authors: Zhu Tongtong , Dhalladoo Subbhalaskshmi , Bibis Sergio , Zufferey Rachel
Published: June 2021

Ether glycerolipid biosynthesis in Leishmania major initiates with the acylation of dihydroxyacetonephosphate by the glycosomal dihydroxyacetonephosphate acyltransferase LmDAT. We previously reported that a null mutant of LmDAT is severely affected in logarithmic growth, survival during stationary phase, and in virulence in mice. In addition, it lacks all ether glycerolipids, produces altered forms of the ether‐lipid based virulence factors lipophosphoglycan and increased levels of GPI‐anchored protein gp63. Here, we describe the characterization of a compensatory mutant of a null strain of LmDAT, Δlmdat/Δlmdatrev. Similarly to the null mutant, the Δlmdat/Δlmdatrev strain formed altered forms of lipophosphoglycan and increased levels of gp63, and was avirulent in mice infection. Further, dihydroxyacetonephosphate acyltransferase activity was absent in the revertant clone, indicating that a mutation in another acyltransferase gene did not confer dihydroxyacetonephosphate specificity. In contrast, the revertant grew normally but still exhibited poor survival during stationary phase. In addition, agarose gel analysis of its genomic DNA failed to detect any amplified DNA. Surprisingly, its sensitivity to aminoglycoside based antibiotic G418 was three times lower than that of the null mutant, while resistance to blasticidin and hygromycin was similar to that of the null mutant.
Effect of Whole Grains on Markers of Systemic Inflammation
Authors: Lefevre Michael , Jonnalagadda Satya S.
Published: June 2021

Reducing subclinical systemic inflammation has been suggested as one potential mechanism to explain the favorable association between whole grain consumption and reduced risk for cardiovascular disease, diabetes and certain cancers. The objective of this study was to review the evidence derived from both epidemiologic and interventions on the effects of whole grain consumption on markers of systemic inflammation. Epidemiological studies provide reasonable support for an association between diets high in whole grains and lower CRP levels. After adjusting for other dietary factors each serving of whole grains is estimated to reduce CRP levels by approximately 7%. This translates to ~20% decline in CRP levels if one achieves the recommended 3 servings/d of whole grains. In contrast to epidemiology studies, intervention studies did not demonstrate a clear effect of increased whole grain consumption on CRP or other markers of inflammation. Issues related to the extent of dietary control, population selection and types of whole grains may underlie these discrepant findings. Based on the existing evidence, it appears that ~40% of the improvement in CRP levels associated with increased whole grain consumption may potentially be through their favorable effect on adiposity and insulin sensitivity. Additional carefully controlled intervention studies are needed to confirm effects of whole grains on subclinical systemic inflammation.
Identifying Nutrition Education Deficits that Affect Food Purchasing by Low‐Income Families with Young Children
Authors: Calloway Eric Elyett , Sweitzer Sara J , Briley Margaret E , Romo Maria , McInnis Katie , Mcallaster Micheal
Published: June 2021

Participants (n=34) were low‐income parents with preschool‐aged children, 72% Hispanic, 91% female, and 75% with total family incomes under $40,000. Parents completed questionnaires concerning knowledge, attitudes, and beliefs regarding family food purchasing habits, and returned food‐purchasing receipts representing one month. Although 88% of parents agreed that consuming diets high in fruits, vegetables and fiber, and low in fat and added sugar can decrease risk for heart disease and diabetes, the knowledge to apply this to their food‐purchasing selections was lacking. Participants could identify foods high in added sugar, but were unable to consistently identify foods high in fat or high in fiber. Out of 98 responses, 60% (n=59) misidentified pasta, bread, or soda as being high in fat. For four items low in dietary fiber (corn flakes, white rice, red meat, and chicken), over 1/3 (n=44) of 130 responses misidentified them as high in dietary fiber and 12% (n=15) indicated “don't know.” Total family food‐expenditure devoted to fruits, vegetables, high‐fiber and low fat food items accounted for only 31% of money spent for food purchases. Though parents agreed healthy diets reduce disease risk, they lacked the knowledge to apply this to food purchasing which presents a significant public health risk and need for nutrition education in vulnerable populations. Funding provided by Little Dudes child care centers.

Grant Funding Source : Little Dudes Child Care Centers
Determining motivators and barriers of college students in purchasing milk from on campus vending for use in development of tailored messages that promote purchase of vended milk
Authors: Sullivan Jennifer , Monnat Laura E , Kennel Julie A , Gunther Carolyn W
Published: June 2021

Calcium intake decreases in many men and women from late adolescence to young adulthood. Because college students rely on food/beverage purchases from vending machines, milk vending, though not widespread, offers an opportunity for improving calcium intake. Research is needed to develop, test, and measure effectiveness of messages that would motivate students to purchase milk from vending machines. The objective of this study was to determine the primary motivators/barriers of on‐campus milk vending purchases in college students for future use in development of tailored messages that promote purchase of vended milk. We surveyed 152 students at the point of purchase from milk and non‐milk beverage vending machines. On a 5‐point Likert scale, the top motivators of consuming vended milk were: ease of taking ‘on the go,’ taste, and variety. Barriers were: mode of purchase, cost, limited availability of milk vending machines on campus, difficulty of taking vended milk ‘on‐the‐go,’ and calorie content. In the next phase of this project, we will develop messages that address these reported motivators/barriers and then test the pre‐determined messages in focus groups of college students. Identifying messages that resonate with the target audience is critical for effectively motivating students to increase consumption of calcium rich foods/beverages, particularly from convenient sources, such as vending machines.
Nutrient intake from food among women of childbearing age ‐ is the burden of nutrient inadequacies growing in the USA?
Authors: Rai Deshanie , Fulgoni Victor L , McBurney Michael , Chapman-Novakofski Karen
Published: June 2021

Currently, we have little understanding of nutrient intakes among women of childbearing age within the U.S.A. This information is relevant given the growing incidence of metabolic complications during pregnancy; accumulating evidence that maternal body weight gain and nutrient intakes prior to and during pregnancy may predict weight gain, body composition and incidence of chronic disease during infancy and childhood. The objective of this study was to evaluate nutrient intakes in women of childbearing age and to ascertain the influence of ethnicity and income. The 2003–2008 National Health and Nutrition Examination Survey data were used to assess nutrient intakes among White (n=1560), African‐American (n=889) and Mexican‐American (n=761) women 19–50 years old. Poverty‐Income Ratio (PIR) was used as an index of family income. African American women had the lowest intake of fiber, folate, vitamin A, calcium and magnesium (p < 0.05). Women (31–50 years) with a PIR of ≤ 1.85 had a significantly lower intake of almost all nutrients analyzed (p < 0.05). Irrespective of ethnicity and income, a significant percentage of the women surveyed were not consuming the recommended amounts of several key nutrients along with fiber. For example, ~80% had vitamin A intake below the RDA and ~70% had calcium intakes below adequate intake. These findings may be a useful resource to guide efforts focusing on nutrition science advocacy and policy in health care settings, government, academia, and private industries.
Health Disparities in Coronary Heart Disease in Miami‐Dade County among Multi‐ethnic populations
Authors: Nagi Gagandeep , Huffman Fatma G.
Published: June 2021

The objective of this study was to measure health disparities in Coronary heart disease (CHD) stratified by race/ethnicity in Miami‐Dade. Data were obtained from Florida Department of Health‐ Bureau of Vital Statistics (N=2,496,435) with 73.8% Whites, 65.0% Hispanics, 18.9% Blacks, 1.5% Asians. Single year rates for Years of potential life lost (YPLL) and Age‐adjusted death rates (AADR) for CHD were used. Disparities were measured in Absolute rate differences and Index of Disparity (ID). Whites were used as the reference point. High absolute rate differences obtained for YPLL in Blacks showed 219.1 more cases per 100,000 population in 2010 than Whites, Hispanics had 1.2 higher cases in 2010 in YPLL than Whites, and Asians had 10.8 lesser cases in 2010 than Whites. Disparity in AADR showed that Asians had 35.6 lesser cases in 2010, and Hispanics had 2.5 times higher cases of mortality in CHD than Whites in 2010. The ID in YPLL for all races as a group was 15.54% whereas AADR showed an ID of 8.69% in 2010. These measurements will be useful for their intended use in communicating the magnitude of disparity and allocating resources towards nutritional assessment and risk prevention for CHD.
Automated DNA extraction and multiplexed PCR assay to identify Escherichia coli O157:H7 from ground beef and kitchen surface swab samples
Authors: Chapman Jessica , Parrish Torrey , Duggan Megan , Williams Patrick
Published: June 2021

Escherichia coli O157:H7 is a common pathogen found in processed beef trimmings and ground beef that has lead to many infectious outbreaks associated with undercooked meat. Only one strain of E. coli is tested for in ground beef, however, legislation is being considered for identifying up to 6 additional strains. We have developed a testing method that uses the Automated Sample Preparation (ASP) instrument, with a multiplexed PCR assay containing HyBeacons, to easily identify three genes in the O157:H7 strain. HyBeacons probes are hybridization probes that simultaneously identify multiple genes via melt curve analysis. In this study, samples of ground beef inoculated with O157:H7, and swabs from inoculated surfaces that had been in contact with ground beef, were cultured at 37°C for 24 hours. Following incubation, 200 μl from each sample was placed into an ASP cartridge for DNA extraction and purification. DNA from each sample was used in a multiplexed PCR assay that identifies stx 1, stx 2, and eae gamma genes of O157:H7. DNA yields from beef culture and swab samples were measured on a NanoDrop (Thermofisher) and yielded an average of 26ng/ul and 17.2 ng/ul respectively. All gene targets were successfully amplified and confirmed via high resolution melt curve analysis. The multiplexed assay would be ideally suited for expansion to include additional O157:H7 genes or even identification of multiple strains of E. coli.
Vitamin D Assessment Using Simple Questionnaires from a College Cohort Study
Authors: Bishop Megan , McDermott Ann Yelmokas , Nazmi Aydin , Stevenson Charles , Hall Laura
Published: June 2021

Vitamin D is important to the health of college students. Our objective was to measure vitamin D intake, sun exposure, skin pigmentation, and serum 25‐hydroxyvitamin D (25[OH]D) in a subset of students from the FLASH Longitudinal College Health Study at Cal Poly, San Luis Obispo, CA. Vitamin D intake and sun exposure were measured using two different tools. First, an online categorical (C) questionnaire administered by the FLASH Study including fish and milk intake, minutes in the sun on the weekdays and weekends. Second, a 28 day recall (R) questionnaire specific to vitamin D foods/supplements, and sun exposure with minutes outside and clothing worn to calculate a sun exposure index (SEI). Skin pigmentation was measured using a reflectance spectrophotometer. Serum 25(OH)D was measured at a local pathology lab. The different measurements for diet and sun exposure were each incorporated into a regression model along with average forehead skin reflectance (FSR) to predict 25(OH)D (Spring 2010, n=35). The average SEI from 11am–3pm (R), fish intake (C), and FSR were significant predictors (R2=0.39, p=0.0036). Weekend sun exposure (C), average monthly intake of vitamin D (R) and FSR were also significant predictors (R2=0.42, p=0.0005). Weekend sun exposure (C), fish intake (C), and FSR were the strongest predictors (R2=0.50, p=0.0001). Thus, simple questionnaires can be used to help determine vitamin D status.
Eating behavior traits during weight loss and weight loss maintenance in premenopausal women
Authors: Zack Melissa K , Shlisky Julie D , Durward Carrie M , Nickols-Richardson Sharon M
Published: June 2021

Changes in body weight (BW), energy intake (EI) and eating behavior (EB) during 12 wks of BW loss and 12 wks of BW loss maintenance were examined. Overweight/obese women (n=56; BMI: 29.8±2.9 kg/m2; age: 35.5±7.2 y) were randomized to 1 of 2 diet plans. BW, EI and EB were measured at wks 0, 12 and 24. In this secondary data analysis, women were grouped by baseline EB as: high restraint/disinhibition (HR/HD); high restraint/low disinhibition (HR/LD); low restraint/high disinhibition (LR/HD); low restraint/disinhibition (LR/LD). Changes in BW, EI and EB were analyzed using repeated measures MANCOVA with pairwise comparisons to detect differences between groups and over time. There was significant BW (kg) loss (p<.001), with no difference between diet groups (p=.89). Over time, energy density (kcal/g) decreased (p<.05), restraint increased (p=.05), and there were interactions between EB group and time for energy (kcal, p<.05), BW (p<.05), BMI (p<.05) and restraint (p<.05). There were differences between EB groups in energy [main effect p<.05, LR/HD>HR/HD p<.05], BW [main effect p<.01, LR/HD>LR/LD p=.01, LR/HD>HR/LD p<.05], BMI [main effect p<.01, LR/HD>LR/LD p<.01] and hunger [main effect p=.001, HR/HD>LR/LD p=.001, HR/HD>HR/LD p<.01]. The influence of EB on EI and BW should be considered during BW loss and BW loss maintenance.

Funded by The Bell Institute of Health and Nutrition, General Mills Inc., Minneapolis, MN.
Anti‐diabetic activities of the extract of Rehmanniae Radix preparata
Authors: Song Jung Eun , You Bo Ram , Kim Min Jee , Kim Mee Ree
Published: June 2021

The present study investigated the anti‐diabetic effects of Rehmanniae Radix preparata extract (RRP) in alloxan‐induced diabetic ICR mouse. Changes of food and water intakes, body weight, blood glucose, insulin, C‐peptide, cholesterol, HDL‐cholesterol, triglyceride, alanine aminotransferase (ALT), aspartate aminotransferase (AST), uric acid (UA), blood urea nitrogen (BUN) in the plasma by administration of RRP(1 g/kg B.W.) were investigated. Body weight gain and food efficiency ratio were significantly lower in diabetic groups than those in normal control group. Concentration of blood glucose was significantly higher in the diabetic control groups than other groups. plasma insulin and c‐peptide level in RRP group was significantly higher than those in diabetes control group (p<0.05), while plasma insulin and c‐peptide level in diabetes group decreased by 59.6% and 67.4%, respectively, compared to those of normal control group. Plasma triglyceride level of diabetes group was significantly higher than that of normal group, whereas that of RRP, group significantly decreased. Plasma HDL‐cholesterol of RRP group was higher than other groups. Meanwhile, the activities of blood plasma AST and UA decreased significantly in RRP group. In conclusion, the results indicated that Rehmanniae Radix preparata improved anti‐diabetic effects by reducing body weight gain and decreasing plasma glucose and triglyceride level.