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Almond consumption and weight loss in obese and overweight adults

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Nut consumption, specifically almonds, have been shown to help maintain weight and influence disease risk factors in adult populations. Limited studies have been conducted examining the effect of a small dose of almonds on energy intake and body weight. The

Nut consumption, specifically almonds, have been shown to help maintain weight and influence disease risk factors in adult populations. Limited studies have been conducted examining the effect of a small dose of almonds on energy intake and body weight. The objective of this study was to determine the influence of pre-meal almond consumption on energy intake and weight in overweight and obese adults. In this study included 21, overweight or obese, participants who were considered healthy or had a controlled disease state. This 8-week parallel arm study, participants were randomized to consume an isocaloric amount of almonds, (1 oz) serving, or two (2 oz) cheese stick serving, 30 minutes before the dinner meal, 5 times per week. Anthropometric measurements including weight, waist circumference, and body fat percentage were recorded at baseline, week 1, 4, and 8. Measurement of energy intake was self-reported for two consecutive days at week 1, 4 and 8 using the ASA24 automated dietary program. The energy intake after 8 weeks of almond consumption was not significantly different when compared to the control group (p=0.965). In addition, body weight was not significantly reduced after 8 weeks of the almond intervention (p=0.562). Other parameters measured in this 8-week trial did not differ between the intervention and the control group. These data presented are underpowered and therefore inconclusive on the effects that 1 oz of almonds, in the diet, 5 per week has on energy intake and bodyweight.

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2011

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Community supported agriculture membership: characterizing food and sustainability behaviors

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Community Supported Agriculture programs (CSAs) have become a viable local source of fresh agricultural goods and represent a potentially new way to improve fruit and vegetable consumption among individuals and families. Studies concerning CSAs have focused mainly on characteristics of

Community Supported Agriculture programs (CSAs) have become a viable local source of fresh agricultural goods and represent a potentially new way to improve fruit and vegetable consumption among individuals and families. Studies concerning CSAs have focused mainly on characteristics of the typical CSA member and motivations and barriers to join a CSA program. The purpose of this study was to examine whether behavior and attitudinal differences existed between current CSA members and a nonmember control group. Specifically, ecological attitudes, eating out behaviors, composting frequency, and family participation in food preparation were assessed. This study utilized an online survey comprising items from previous survey research as well as newly created items. A total of 115 CSA member and 233 control survey responses were collected. CSA members were more likely to be older, have more education, and have a higher income than the control group. The majority of CSA members surveyed were female, identified as non-Hispanic and Caucasian, earned a higher income, and reported being the primary food shopper and preparer. The majority of members also noted that the amount and variety of fruits and vegetables they ate and served their family increased as a result of joining a CSA. CSA members were more ecologically minded compared to the control group. Frequency of eating out was not significantly different between groups. However, eating out behaviors were different between income categories. CSA members spent significantly more money at each meal eaten away from home and spent significantly more money on eating out each week. In both cases, controlling for income attenuated differences between groups. CSA members composted at a significantly higher rate and took part in other eco-friendly behaviors more often than the control group. Finally, no significant difference was evident between the two groups when analyzing family involvement in food preparation and meal decision-making. Overall, some significant attitudinal and behavioral differences existed between CSA members and non-CSA members. Further research is necessary to examine other distinctions between the two groups and whether these differences occur as a result of CSA membership.

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2011

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Synthesis and evaluation of a new class of cancer chemotherapeutics based on purine-like extended amidines

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A potential new class of cancer chemotherapeutic agents has been synthesized by varying the 2 position of a benzimidazole based extended amidine. Compounds 6-amino-2-chloromethyl-4-imino-1-(2-methansulfonoxyethyl)-5-methyl-1H-benzimidazole-7-one (1A) and 6-amino-2-hydroxypropyl-4-imino-1-(2-methansulfonoxyethyl)-5-methyl-1H-benzimidazole-7-one (1B) were assayed at the National Cancer Institute's (NCI) Developmental Therapeutic Program (DTP)

A potential new class of cancer chemotherapeutic agents has been synthesized by varying the 2 position of a benzimidazole based extended amidine. Compounds 6-amino-2-chloromethyl-4-imino-1-(2-methansulfonoxyethyl)-5-methyl-1H-benzimidazole-7-one (1A) and 6-amino-2-hydroxypropyl-4-imino-1-(2-methansulfonoxyethyl)-5-methyl-1H-benzimidazole-7-one (1B) were assayed at the National Cancer Institute's (NCI) Developmental Therapeutic Program (DTP) and found to be cytotoxic at sub-micromolar concentrations, and have shown between a 100 and a 1000-fold increase in specificity towards lung, colon, CNS, and melanoma cell lines. These ATP mimics have been found to correlate with sequestosome 1 (SQSTM1), a protein implicated in drug resistance and cell survival in various cancer cell lines. Using the DTP COMPARE algorithm, compounds 1A and 1B were shown to correlate to each other at 77%, but failed to correlate with other benzimidazole based extended amidines previously synthesized in this laboratory suggesting they operate through a different biological mechanism.

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2011

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The indices of bone changes in response to exercise

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The gold standard for bone measurement is DXA (dual energy X-ray absorptiometry). Typically, to observe changes in bone by DXA, a minimum of a 4-month intervention is required. Serum osteocalcin (OST) (a bone formation marker) and quantitative ultrasound (QUS) of

The gold standard for bone measurement is DXA (dual energy X-ray absorptiometry). Typically, to observe changes in bone by DXA, a minimum of a 4-month intervention is required. Serum osteocalcin (OST) (a bone formation marker) and quantitative ultrasound (QUS) of the calcaneus can be used as indicators of bone change but the sensitivity and time course of these indices to short term interventions are unknown. The purpose of this study was twofold: to compare monthly changes in OST and QUS in response to jump training and to evaluate the relationship between DXA, OST and QUS. Young women with QUS t-scores less than 1.0 were randomized into a jump training (J) (n=16) or control (C) (n=16). J consisted of a progressive routine of 1 and 2-footed jumping performed 3 days per week for 4 months. Body composition, QUS and OST were measured at baseline, and monthly for 4 months. DXA and 24-hour dietary recalls were completed at baseline and 4 months. Low attrition rate (12.5%) and high compliance (98%) with the exercise intervention was recorded. No significant correlations between QUS and OST existed. No significant differences were observed between groups at baseline in body composition or bone variables. Monthly increases in OST were observed but there were no significant differences over time between groups in any bone variables. OST and QUS may be indicative of short term bone changes but these variables were not specifically sensitive to the jumping intervention in this population of women.

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2011

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Mapping the RNA-protein interface in telomerase RNP

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In the 1970s James Watson recognized the inability of conventional DNA replication machinery to replicate the extreme termini of chromosomes known as telomeres. This inability is due to the requirement of a building block primer and was termed the

In the 1970s James Watson recognized the inability of conventional DNA replication machinery to replicate the extreme termini of chromosomes known as telomeres. This inability is due to the requirement of a building block primer and was termed the end replication problem. Telomerase is nature's answer to the end replication problem. Telomerase is a ribonucleoprotein which extends telomeres through reverse transcriptase activity by reiteratively copying a short intrinsic RNA sequence to generate 3' telomeric extensions. Telomeres protect chromosomes from erosion of coding genes during replication, as well as differentiate native chromosome ends from double stranded breaks. However, controlled erosion of telomeres functions as a naturally occurring molecular clock limiting the replicative capacity of cells. Telomerase is over activated in many cancers, while inactivation leads to multiple lifespan limiting human diseases. In order to further study the interaction between telomerase RNA (TR) and telomerase reverse transcriptase protein (TERT), vertebrate TERT fragments were screened for solubility and purity following bacterial expression. Soluble fragments of medaka TERT including the RNA binding domain (TRBD) were identified. Recombinant medaka TRBD binds specifically to telomerase RNA CR4/CR5 region. Ribonucleotide and amino acid pairs in close proximity within the medaka telomerase RNA-protein complex were identified using photo-activated cross-linking in conjunction with mass spectrometry. The identified cross-linking amino acids were mapped on known crystal structures of TERTs to reveal the RNA interaction interface of TRBD. The identification of this RNA TERT interaction interface furthers the understanding of the telomerase complex at a molecular level and could be used for the targeted interruption of the telomerase complex as a potential cancer treatment.

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2011

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Almond consumption and dietary compensation in overweight and obese adults

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ABSTRACT Epidemiological studies have suggested a link between nut consumption and weight. The possible effects of regular nut consumption as a method of weight loss has shown minimal results with 2-3 servings of nut products per day. This 8 week

ABSTRACT Epidemiological studies have suggested a link between nut consumption and weight. The possible effects of regular nut consumption as a method of weight loss has shown minimal results with 2-3 servings of nut products per day. This 8 week study sought to investigate the effect of more modest nut consumption (1 oz./day, 5 days/week) on dietary compensation in healthy overweight individuals. Overweight and obese participants (n = 28) were recruited from the local community and were randomly assigned to either almond (NUT) or control (CON) group in this randomized, parallel-arm study. Subjects were instructed to eat their respective foods 30 minutes before the dinner meal. 24 hour diet recalls were completed pre-trial and at study weeks 1, 4 and 8. Self-reported satiety data were completed at study weeks 1, 4, and 8. Attrition was unexpectedly high, with 13 participants completing 24 dietary recall data through study week 8. High attrition limited statistical analyses. Results suggested a lack of effect for time or interaction for satiety data (within groups p = 0.997, between groups p = 0.367). Homogeneity of of inter-correlations could not be tested for 24-hour recall data as there were fewer than 2 nonsingular cell covariance matrices. In conclusion, this study was unable to prove or disprove the effectiveness of almonds to induce dietary compensation.

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2011

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Novel strategies for producing proteins with non-proteinogenic amino acids

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The biological and chemical diversity of protein structure and function can be greatly expanded by position-specific incorporation of non-natural amino acids bearing a variety of functional groups. Non-cognate amino acids can be incorporated into proteins at specific sites by using

The biological and chemical diversity of protein structure and function can be greatly expanded by position-specific incorporation of non-natural amino acids bearing a variety of functional groups. Non-cognate amino acids can be incorporated into proteins at specific sites by using orthogonal aminoacyl-tRNA synthetase/tRNA pairs in conjunction with nonsense, rare, or 4-bp codons. There has been considerable progress in developing new types of amino acids, in identifying novel methods of tRNA aminoacylation, and in expanding the genetic code to direct their position. Chemical aminoacylation of tRNAs is accomplished by acylation and ligation of a dinucleotide (pdCpA) to the 3'-terminus of truncated tRNA. This strategy allows the incorporation of a wide range of natural and unnatural amino acids into pre-determined sites, thereby facilitating the study of structure-function relationships in proteins and allowing the investigation of their biological, biochemical and biophysical properties. Described in Chapter 1 is the current methodology for synthesizing aminoacylated suppressor tRNAs. Aminoacylated suppressor tRNACUAs are typically prepared by linking pre-aminoacylated dinucleotides (aminoacyl-pdCpAs) to 74 nucleotide (nt) truncated tRNAs (tRNA-COH) via a T4 RNA ligase mediated reaction. Alternatively, there is another route outlined in Chapter 1 that utilizes a different pre-aminoacylated dinucleotide, AppA. This dinucleotide has been shown to be a suitable substrate for T4 RNA ligase mediated coupling with abbreviated tRNA-COHs for production of 76 nt aminoacyl-tRNACUAs. The synthesized suppressor tRNAs have been shown to participate in protein synthesis in vitro, in an S30 (E. coli) coupled transcription-translation system in which there is a UAG codon in the mRNA at the position corresponding to Val10. Chapter 2 describes the synthesis of two non-proteinogenic amino acids, L-thiothreonine and L-allo-thiothreonine, and their incorporation into predetermined positions of a catalytically competent dihydrofolate reductase (DHFR) analogue lacking cysteine. Here, the elaborated proteins were site-specifically derivitized with a fluorophore at the thiothreonine residue. The synthesis and incorporation of phosphorotyrosine derivatives into DHFR is illustrated in Chapter 3. Three different phosphorylated tyrosine derivatives were prepared: bis-nitrobenzylphosphoro-L-tyrosine, nitrobenzylphosphoro-L-tyrosine, and phosphoro-L-tyrosine. Their ability to participate in a protein synthesis system was also evaluated.

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2013

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The efficacy of nopales (Opuntia spp) on lipoprotein profile and oxidative stress among moderately hypercholesterolemic adults

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Background: Evidence about the purported hypoglycemic and hypolipidemic effects of nopales (prickly pear cactus pads) is limited. Objective: To evaluate the efficacy of nopales for improving cardiometabolic risk factors and oxidative stress, compared to control, in adults with hypercholesterolemia. Design:

Background: Evidence about the purported hypoglycemic and hypolipidemic effects of nopales (prickly pear cactus pads) is limited. Objective: To evaluate the efficacy of nopales for improving cardiometabolic risk factors and oxidative stress, compared to control, in adults with hypercholesterolemia. Design: In a randomized crossover trial, participants were assigned to a 2-wk intervention with 2 cups/day of nopales or cucumbers (control), with a 2 to 3-wk washout period. The study included 16 adults (5 male; 46±14 y; BMI = 31.4±5.7 kg/m2) with moderate hypercholesterolemia (low density lipoprotein cholesterol [LDL-c] = 137±21 mg/dL), but otherwise healthy. Main outcomes measured included: dietary intake (energy, macronutrients and micronutrients), cardiometabolic risk markers (total cholesterol, LDL-c, high density lipoprotein cholesterol [HDL-c], triglycerides, cholesterol distribution in LDL and HDL subfractions, glucose, insulin, homeostasis model assessment, and C-reactive protein), and oxidative stress markers (vitamin C, total antioxidant capacity, oxidized LDL, and LDL susceptibility to oxidation). Effects of treatment, time, or interactions were assessed using repeated measures ANOVA. Results: There was no significant treatment-by-time effect for any dietary composition data, lipid profile, cardiometabolic outcomes, or oxidative stress markers. A significant time effect was observed for energy, which was decreased in both treatments (cucumber, -8.3%; nopales, -10.1%; pTime=0.026) mostly due to lower mono and polyunsaturated fatty acids intake (pTime=0.023 and pTime=0.003, respectively). Both treatments significantly increased triglyceride concentrations (cucumber, 14.8%; nopales, 15.2%; pTime=0.020). Despite the lack of significant treatment-by-time effects, great individual response variability was observed for all outcomes. After the cucumber and nopales phases, a decrease in LDL-c was observed in 44% and 63% of the participants respectively. On average LDL-c was decreased by 2.0 mg/dL (-1.4%) after the cucumber phase and 3.9 mg/dL (-2.9%) after the nopales phase (pTime=0.176). Pro-atherogenic changes in HDL subfractions were observed in both interventions over time, by decreasing the proportion of HDL-c in large HDL (cucumber, -5.1%; nopales, -5.9%; pTime=0.021) and increasing the proportion in small HDL (cucumber, 4.1%; nopales, 7.9%; pTime=0.002). Conclusions: These data do not support the purported benefits of nopales at doses of 2 cups/day for 2-wk on markers of lipoprotein profile, cardiometabolic risk, and oxidative stress in hypercholesterolemic adults.

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2013

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Studies on the three-dimensional structures of proteins using X-ray crystallography

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X-ray diffraction is the technique of choice to determine the three-dimensional structures of proteins. In this study it has been applied to solve the structure of the survival motor neuron (SMN) proteins, the Fenna-Mathews-Olson (FMO) from Pelodictyon phaeum (Pld. phaeum)

X-ray diffraction is the technique of choice to determine the three-dimensional structures of proteins. In this study it has been applied to solve the structure of the survival motor neuron (SMN) proteins, the Fenna-Mathews-Olson (FMO) from Pelodictyon phaeum (Pld. phaeum) protein, and the synthetic ATP binding protein DX. Spinal muscular atrophy (SMA) is an autosomal recessive genetic disease resulting in muscle atrophy and paralysis via degeneration of motor neurons in the spinal cord. In this work, we used X-ray diffraction technique to solve the structures of the three variant of the of SMN protein, namely SMN 1-4, SMN-WT, and SMN-Δ7. The SMN 1-4, SMN-WT, and SMN-Δ7 crystals were diffracted to 2.7 Å, 5.5 Å and 3.0 Å, respectively. The three-dimensional structures of the three SMN proteins have been solved. The FMO protein from Pld. phaeum is a water soluble protein that is embedded in the cytoplasmic membrane and serves as an energy transfer funnel between the chlorosome and the reaction center. The FMO crystal diffracted to 1.99Å resolution and the three-dimensional structure has been solved. In previous studies, double mutant, DX, protein was purified and crystallized in the presence of ATP (Simmons et al., 2010; Smith et al. 2007). DX is a synthetic ATP binding protein which resulting from a random selection of DNA library. In this study, DX protein was purified and crystallized without the presence of ATP to investigate the conformational change in DX structure. The crystals of DX were diffracted to 2.5 Å and the three-dimensional structure of DX has been solved.

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2013

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Directed evolution of gp120 binding mutants of the lectin Cyanovirin-N

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Cyanovirin-N (CV-N) is a naturally occurring lectin originally isolated from the cyanobacteria Nostoc ellipsosporum. This 11 kDa lectin is 101 amino acids long with two binding sites, one at each end of the protein. CV-N specifically binds to terminal Manα1-2Manα

Cyanovirin-N (CV-N) is a naturally occurring lectin originally isolated from the cyanobacteria Nostoc ellipsosporum. This 11 kDa lectin is 101 amino acids long with two binding sites, one at each end of the protein. CV-N specifically binds to terminal Manα1-2Manα motifs on the branched, high mannose Man9 and Man8 glycosylations found on enveloped viruses including Ebola, Influenza, and HIV. wt-CVN has micromolar binding to soluble Manα1-2Manα and also inhibits HIV entry at low nanomolar concentrations. CV-N's high affinity and specificity for Manα1-2Manα makes it an excellent lectin to study for its glycan-specific properties. The long-term aim of this project is to make a variety of mutant CV-Ns to specifically bind other glycan targets. Such a set of lectins may be used as screening reagents to identify biomarkers and other glycan motifs of interest. As proof of concept, a T7 phage display library was constructed using P51G-m4-CVN genes mutated at positions 41, 44, 52, 53, 56, 74, and 76 in binding Domain B. Five CV-N mutants were selected from the library and expressed in BL21(DE3) E. coli. Two of the mutants, SSDGLQQ-P51Gm4-CVN and AAGRLSK-P51Gm4-CVN, were sufficiently stable for characterization and were examined by CD, Tm, ELISA, and glycan array. Both proteins have CD minima at approximately 213 nm, indicating largely β-sheet structure, and have Tm values greater than 40°C. ELISA against gp120 and RNase B demonstrate both proteins' ability to bind high mannose glycans. To more specifically determine the binding specificity of each protein, AAGRLSK-P51Gm4-CVN, SSDGLQQ-P51Gm4-CVN, wt-CVN, and P51G-m4-CVN were sent to the Consortium for Functional Glycomics (CFG) for glycan array analysis. AAGRLSK-P51Gm4-CVN, wt-CVN, and P51G-m4-CVN, have identical specificities for high mannose glycans containing terminal Manα1-2Manα. SSDGLQQ-P51Gm4-CVN binds to terminal GlcNAcα1-4Gal motifs and a subgroup of high mannose glycans bound by P51G-m4-CVN. SSDGLQQ-wt-CVN was produced to restore anti-HIV activity and has a high nanomolar EC50 value compared to wt-CVN's low nanomolar activity. Overall, these experiments show that CV-N Domain B can be mutated and retain specificity identical to wt-CVN or acquire new glycan specificities. This first generation information can be used to produce glycan-specific lectins for a variety of applications.

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2013