bluuebunny/crossref_metadata_2025 huggingface dataset:
Hydrogen peroxide effects upon viability and oxygen consumption by Kluyveromyces marxianus and Saccharomyces cerevisiae wild yeast
Authors: Arellano‐Plaza Melchor , Noriega‐Cisneros Ruth , Estrada‐Villagómez M. Mirella , Clemente‐Guerrero Mónica , Manzo‐Avalos Salvador , González‐Hernández Juan Carlos , Saavedra‐Molina Alfredo
Published: June 2021

The production of reactive oxygen species (ROS) are generated during normal cellular metabolism and originated from the presence of pro‐oxidants such as H2O2 or the exposure to ionizing radiations. The ROS damage cellular components by oxidizing lipids, proteins and nucleic acids. The aim of this work was to study the effect of the H2O2 during the stationary phase (24, 48, 72 and 96 h) to wild yeasts (two Kluyveromyces marxianus [SLPA and OFF1] and one Saccharomyces cerevisiae [MC4], all of them were obtained from mezcal processes). The exposure for 1 h to hydrogen peroxide of the wild yeast grown on YPD, affected the cellular viability at the beginning of the stationary phase (24 h of growth) decreased almost 100% the cellular viability independently of the pro‐oxidant concentration in SLPA and OFF1, in the MC4 decreased only 30%. The viability increased in all the yeast after 48 h of growth, the MC4 viability were 80% at 90 mM and 100% at 10 mM. The OFF1 increased the viability only to 60% after 96 h of growth at 10 mM and 25% at 90 mM. The oxygen consumption rate decreased when increased H2O2 concentration and the growth time, obtained no consumption at 96 h of growth and 90 mM but the viability were almost 100% in SLPA. These results suggest the resistance to the H2O2 by wild yeast during the stationary phase, but the oxygen consumption was inhibited.
Mechanism of ATP‐dependent release of wild type and mutant human brain hexokinases from mitochondria
Authors: Mehyar Nimer , Watanabe Muneaki , Shen Lu , Skaff Andrew , Honzatko Richard B
Published: June 2021

Adenosine 5′‐triphosphate (ATP) can release hexokinase I (HKI) from its binding site on outer mitochondrial membrane. The mechanism of ATP release of HKI from mitochondria, however, is not clear: ATP binds to the C‐terminal half of HKI as a substrate and to the N‐terminal half near the membrane binding element. ATP also binds to voltage dependent anion channel (VDAC), the integral membrane component that binds HKI to the mitochondrion. Fluorescent nucleotide analogue 2′,3′‐O‐(2,4,6‐trinitrophenyl) adenosine 5′‐diphosphate (TNP‐ADP) binds with high affinity to the active site of HKI and to VDAC, but not to the N‐terminal half of HKI. ATP and TNP‐ADP separately release wild type HKI and the truncated N‐domain of HKI from mitochondria, excluding nucleotide binding to either the N‐ or C‐half of HKI in the release mechanism. Results here are consistent with a HKI release mechanism in which ATP or TNP‐ADP bind to VDAC. This research is supported by NIH grant NS 10546.
Oxidation‐ or Radiation‐Derived Effects on Human Mitochondria as Seen in a Fibroblast Cell Line Derived from a Family Known to Have Elevated Levels of the 4977‐bp Common Deletion in its Mitochondrial DNA
Authors: Stoll Zachary , Pogozelski Wendy
Published: June 2021

DNA in the mitochondria of cells undergoes deletion events that compromise energy function but do not prevent cells from being viable. The mechanism of how these deletions occur and how they affect cell function are topics of interest. We have been investigating the role and induction of large‐scale deletions in oxidative stress using a variety of methods, including gamma radiation. We observed a dose‐dependent increase in the levels of a 4977‐bp deletion called the “common deletion.” using real‐time PCR with an external DNA quantification standard in fibroblast cells. We are currently manipulating cell conditions to alter the metabolism of cells and compare the effects with and without this deletion.

This research was sponsored by the Camille and Henry Dreyfus Foundation, the Geneseo Foundation and the Geneseo Student Association.
Functional expression and characterization of CYP51 from Candida albicans
Authors: Park Hyoung‐Goo , Han Songhee , Lim Young‐Ran , Eun Chang‐Yong , Kim Donghak
Published: June 2021

An opportunistic fungal pathogen, Candida albicans contains the 10 putative cytochrome P450 (CYP) genes. They seem to play important roles for fungal survival and virulence. Here, we report the identification and characterization of CYP51, a putative lanosterol demethylase. The recombinant CYP51 protein containing a 6×(His)‐tag has been expressed in Escherichia coli and purified. The purified proteins showed the tight binding spectra with lanosterol. It exhibited a lanosterol demethylase activity in reconstitution with rat P450‐NADPH reductase to produce de methylated product in HPLC analysis. Azole compounds affinities of CYP51 enzyme was calibrated with econazole, itraconazole, ketoconazole and fluconazole. All four azoles showed Type II spectra with tight binding affinities to C. albicans CYP51. This study of functional characterization of CYP51 from C. albicans could provide the critical understanding of its role in the processing of pathogenesis. This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2009‐0088150).
Global Protein Synthesis and Hypoxia Responses in Yeast
Authors: Cadinu Daniela , Shah Ajit N. , Dastidar Ranita Ghosh , Henke R. Michael , Zhang Li
Published: June 2021

Oxygen is critical for the survival and development of many living organisms, particularly eukaryotes. Living organisms ranging from yeast to humans have developed sophisticated mechanisms to respond to the changes of oxygen level in the environment. We used the yeast Saccharomyces cerevisiae as a model system to study mechanisms of oxygen sensing and regulation. To identify potential global regulators mediating oxygen regulation and hypoxia responses, we initiated a genome‐wide screen for deletion mutants that enable cells to grow better under hypoxia. By visual screening, we initially identified 104 yeast deletion mutants that grew significantly better under hypoxia, compared to the wild type parent from a library of 4786 yeast deletion strains. Further quantitative measurements showed that six deletion mutants of genes encoding functions involved in ribosome biogenesis exhibit the highest growth enhancement under hypoxia. Experiments are underway to investigate the molecular mechanisms by which ribosome biogenesis and global protein synthesis promote cell growth under hypoxia. GM062246
Cyclic AMP‐independent phosphoregulation of protein kinase A in the fission yeast, Schizosaccharomyces pombe
Authors: McInnis Brittney , Marcus Stevan
Published: June 2021

In the fission yeast, Schizosaccharomyces pombe, cyclic AMP (cAMP) and cAMP‐dependent protein kinase (PKA) are not essential for cell viability under normal culturing conditions, making this organism attractive for investigating cAMP‐independent mechanisms of PKA regulation. Here we show that S. pombe adenylate cyclase (cyr1Δ) mutants express significantly higher levels of the PKA catalytic subunit, Pka1, than wild type S. pombe cells, while expressing similar levels of the PKA regulatory subunit, Cgs1. Furthermore, in cyr1Δ cells, but not wild type cells, a substantial proportion of Pka1 protein is hyperphosphorylated by a mechanism dependent on the PKA regulatory subunit, Cgs1. Hyperphosphorylation of Pka1 is further induced in cyr1Δ cells, and to varying degrees in wild type S. pombe cells, in response to KCl stress, glucose derepression, and stationary phase growth. Our findings demonstrate the existence of a cAMP‐independent mechanism(s) of PKA phosphoregulation in S. pombe, which we speculate may function to maintain certain PKA functions in conditions in which cAMP‐dependent activities are downregulated.
Covalent Immobilization of Carbonic Anhydrase into Mesoporous Silica Nanoparticles for Drug Delivery Applications
Authors: Mendez Jessica , Griebenow Kai
Published: June 2021

The overall goal of this research project is to develop a novel nanoparticle drug delivery system through the covalent immobilization of a bio‐therapeutic agent into mesoporous silica nanoparticles (MSN). We activated the MSN through its functionalization with thiol groups. Carbonic anhydrase (CA) was modified with a cross‐linker, which will allow its immobilization into the carrier. Characterization of the activated surface was performed utilizing XPS, DLS, and SEM. We determined the level of modification for the enzyme and studied the impact of the modification on its residual activity and tertiary structure. The results demonstrated that the MSN were functionalized successfully and that the activation process decreased the size of the particles, but did not impact their morphology. Residual enzyme activity after modification was determined to be 76% and 50% for a 6% and 19% degree of modification, respectively. The CD spectra revealed that not significant changes in the tertiary structure occurred due to enzyme activation. Subsequently, the enzyme was covalently immobilized into the MSN and experiments related to establish the maximum amount of enzyme that can be immobilized into the thiol‐functionalized MSN were conducted. The release profile of the immobilized enzyme at physiological conditions was determined. Future experiments will include characterization of the released enzyme from the MSN.
Endocytosis of pro‐apoptotic twin‐base RGDSK helical rosette nanotubes in human U937 macrophage cell line
Authors: Singh (Suri) Sarabjeet , Fenniri Hicham , Hemraz Usha , Singh Baljit
Published: June 2021

Twin‐base helical rosette nanotubes (RNTs), a novel class of biologically inspired nanotubes, hold tremendous potential as targeted drug delivery shuttle. To realize biomedical potential of RNT, there is a critical need to understand the interaction of RNT with the cells. Therefore, we synthesized hybrid RNT that composed of RGDSK and FITC to gain ability to track them during in vitro experiments with human differentiated macrophages. Scanning electron microscopy confirmed nano‐scale dimensions of RGDSK RNTs. FITC‐RGDSK RNT was co‐localized with integrin avb3‐Cy5 on the macrophages within 2 minutes of incubation at 37 deg C. Integrin avb3 and flotallin‐1, a lipid rafts protein, showed fever co‐localization in non‐stimulated cells. FITC‐RGDSK RNT also co‐localizes with EEA1, an early endosome marker. The inhibition of the co‐localization of FITC‐RGDSK RNT with the integrin and translocation into endosomes at 4 deg C demonstrated energy‐dependence of the process. The incubation of differentiated macrophages with RGDSK‐TBL RNT (>5 μM) resulted in nearly 100 % apoptosis within 12 hours. These results suggest that the FITC/RGDSK‐TBL RNT could be used as a versatile platform to deliver a variety of biologically active molecules for cancer therapy.

Support: NSERC, AARI.

Grant Funding Source: AARI, NSERC
Microbiological evaluation of 3‐((E)‐(4‐chloro‐6‐hydroxypyrimidin‐2‐ylimino)methyl)‐4‐chloro‐2H‐chromen‐2‐one
Authors: Govori Sevdije R , Spahiu Sabrije , Haziri Arben , Daci Majlinda , Sehabi Muhamet , Kurteshi Kemajl
Published: June 2021

3‐((E)‐(4‐chloro‐6‐hydroxypyrimidin‐2‐ylimino)methyl)‐4‐chloro‐2H‐chromen‐2‐one 3 is synthesized in very good yields by reaction of corresponding 4‐chloro‐2‐oxo‐2H‐chromene‐3‐carbaldehyde 1 and 2‐amino‐6‐chloropyrimidin‐4‐ol 2. Antibacterial activity of products 3 is investigated and results are submitted for their activities against Escherichia coli and Enterochoaco. Applying the disc method we mesasured diameters of the inhibition zone around disc which are previously wetted with N,N‐DMF solution of compounds 3 in concentration 1 mg/l, 0.5 mg/l and 0.1 mg/l.

The compounds 3 showed strong activity in reducing the microbial growth. The synthesized compound 3 showed most effective antibacterial activity against Escherichia coli.
GM‐CSF plays a key role in zymosan‐stimulated human dendritic cells for activation of Th1 and Th17 cells
Authors: Wei Wen‐Chi , Su Yi‐Hsuan , Staniforth Vanisree , Sheu Jyh‐Horng , Yang Ning‐Sun
Published: June 2021

In this study, we evaluate comparatively the effect of zymosan and LPS on human monocyte‐derived dendritic cells (hDCs). The specific and differential effects of zymosan on the expression of several key cytokines, including granulocyte‐macrophage colony stimulating factor (GM‐CSF), interleukins (IL‐1£\, IL‐1£] and IL‐10) and monocyte chemoattractant protein‐1 (MCP‐1), were found to be distinctive from the effects of LPS. Unlike with LPS activation, DCs under zymosan activation expressed little or no IL‐12 p70 due to the lack of expression of the p35 subunit. However, treatment with zymosan resulted in a substantial increase in Th1 and Th17 cell polarizing capacities. Furthermore, GM‐CSF secreted by zymosan activated DCs enhanced IL‐23 production, subsequently resulting in activation of Th17 response. GM‐CSF and IL‐27, rather than IL‐12 p70, were both major direct contributors to the activation of Th1 response. This mechanism is drastically different or even opposite, but complementary, to the LPS‐mediated T‐cell activation. Therefore, we suggest that this novel, zymosan‐induced, GM‐CSF‐mediated signaling network may be employed as a future guide for modulation and differentiation of specific immune cell type activities.
Development of a laboratory skills survey and assessment tool for use in the biochemistry teaching laboratory
Authors: Caldwell Benjamin D , Daggett Melissa
Published: June 2021

For the past three years we have been examining student learning of laboratory skills in an introductory biochemistry course. Our focus has been on evaluating student performance of basic lab skills and comprehension of the theories behind the techniques and methods used in the biochemistry laboratory. Some topics and skills that have examined include the use of basic mathematic skills in the laboratory (i.e. preparing solutions and buffers, making dilutions, and determining accuracy and precision of measurements), mastery of basic laboratory equipment, (micropipettes, pH meters and balances) and the use of instruments such as spectrophotometers (UV/Vis and fluorimeters) used for protein assays and enzyme kinetics. As an addition component we have paired a personal self‐assessment survey for measuring students’ opinions of their understanding of these basic labs skills with a quantitative test of their ability to use data. This presentation will discuss the development of these tools in measuring student performance and comprehension of essential biochemistry lab skills.
Using assessment data to direct instructional practices in the biochemistry classroom
Authors: Loertscher Jennifer , Minderhout Vicky , Lewis Jennifer , Villafane Sachel
Published: June 2021

Recent recommendations by professional societies and federal scientific agencies encourage college and university instructors to consider making changes in the content and structure of their biochemistry courses. Although much evidence exists to support the use of active learning strategies in the classroom, less is known about specific content knowledge that students bring from prior coursework into the biochemistry classroom. Prior knowledge is both expected and built upon in biochemistry courses. We are currently developing an instrument to measure student retention of general chemistry and general biology knowledge in key concept areas. The areas chosen for study are associated with common student misconceptions that chemistry and biology faculty have observed. While our instrument is in early stages of development, we are able to draw some conclusions about the preparedness of students in discrete aspects of prior knowledge. Specifically, preliminary data give new insight into student understanding of foundational concepts including bond energy, intermolecular forces, pKa, free energy changes, and protein structure. Data resulting from this study could guide curricular reform in the future.
Protein Analysis Using Micro‐Electrophoresis and Micro‐Spectroscopy
Authors: Whitson Matthew McGuire , Mancuso Matthew Cian , Harwood John J , Boles Jeffrey Oakley
Published: June 2021

Since the development of the Bradford and Lowry protein assays in 1976 and 1951, respectively, many additional techniques for the determination of protein concentration have been developed based on dye‐binding technologies. In an effort to exploit current available technologies and at the same time expose undergraduate and graduate students to emerging techniques, we carried out a comparative analysis using both traditional methods and methods exploiting micro‐electrophoresis and micro‐spectroscopy. We use a bioanalyzer “lab‐on‐a‐chip” instrument in the analysis, which represents an example of a micro‐total analysis system (u‐TAS). We are developing experiments to help chemistry students in their junior year become more familiar with biotechnology instrumentation that can be used in undergraduate and graduate research. Our poster will present the theoretical principles demonstrated in these teaching experiments, and practical details on how to include both micro‐electrophoresis and micro‐spectroscopy in chemistry and biochemistry courses. This research has been supported by the National Science Foundation.
MULTIDISCIPLINARY BIOLOGY TRAINING ‐ Undergraduate Student Perspective
Authors: Candas Mehmet , Sessoms Rebekah , Lebens Kaylen
Published: June 2021

Majority of undergraduate students in biology prospect themselves toward a career in medical professions. However, biology is a vibrant discipline and a degree in biology prepares for careers in many other professions with opportunities in a wide range of fields. Therefore, it is imperative that undergraduate biology students are prepared to recognize the potential of biology background. A key factor in this preparation is an extensive curriculum that integrates contemporary, multidisciplinary concepts so that graduates become aware of issues in a number of fields and feel empowered with their knowledge. This also is critical to building student confidence and eliminating postgraduation uncertainty. Another key factor is the engagement of students in laboratory activities. Hands‐on technical exercises cultivate students’ capacity to work cooperatively, multitask, manage time and resources, and think about how concepts and applications are related. Finally, students feel stimulated when collaborations and interactions between students are nurtured since such an educational setting generates positive experience with greater sense of belonging. In conclusion, it is important that undergraduate biology students acknowledge the broad prospects of the bachelor's degrees in biology and the importance of multidisciplinary training in realizing the potential of biology in the 21st century.
The action of vitamin K in brain during aging is linked to the pAKT/AKT signaling pathway
Authors: Guylaine Ferland , Mabit Alexandra , Fournier Chantal , Letourneau Jacynthe
Published: June 2021

Vitamin K (VK) participates in brain function through sphingolipid metabolism and activation of protein Gas6. We have provided evidence that long‐term low phylloquinone (K1) intake leads to cognitive deficits in aging rats and to a concurrent accumulation of ceramides in certain brain regions. To gain further insight on these findings, we investigated the pAKT/AKT cell signaling and caspase‐3 apoptotic processes in 20 mo‐old female SD rats which had been fed diets containing either very low (VL: 80), low (L: 500) and moderately high (H: 2000) levels of K1 (μg/Kg diet; n=5/group). A fourth group fed the L diet until 14 mo and the H diet until sacrifice was also investigated. The pAKT/AKT ratio (western blot) and caspase‐3 activity (enzymatic assay) which reflect cell survival and cell death respectively, were assessed in pons medulla, frontal cortex, striatum and hippocampus. Strong trends were observed for higher cell survival in the striatum and frontal cortex of rats from the H group. Furthermore, when VK was fed in limited amounts (groups VL and L), pAKT/AKT ratios were higher in the hippocampus than in the other regions. In contrast, caspase‐3 activity was more heterogenous, a finding which may be linked to the assay used. In conclusion, long‐term consumption of a VK rich diet appears to offer cellular protection in brain and could contribute to the maintenance of cognitive functions during aging. Supported by CIHR.
γ‐Tocopherol supplementation improves postprandial vascular endothelial function in lean and obese men by decreasing oxidative and nitrative stress
Authors: Mah Eunice , Ballard Kevin D , Kawiecki Diana M , Volek Jeff S , Bruno Richard S
Published: June 2021

Hyperglycemia induces oxidative stress and inflammation that impairs vascular endothelial function (VEF). We hypothesized that γ‐tocopherol (γ‐T) supplementation would improve hyperglycemia‐mediated decreases in VEF by reducing oxidative stress and inflammatory responses. A randomized, crossover study was conducted in lean and obese men (n=8/group) who followed their usual diet or were also provided γ‐T‐rich capsules (500 mg/d, 5 d) prior to an oral glucose challenge. Brachial artery flow‐mediated dilation (FMD) and markers of oxidative/nitrative stress and inflammation were measured throughout the 3 h postprandial period. Supplementation increased plasma γ‐T by 170% in all men without affecting α‐T. Baseline HOMA was greater in obese men and correlated to FMD, but was unchanged by γ‐T. Postprandial hyperglycemia decreased FMD (p<0.05) in both groups. Postprandial FMD area under the curve (AUC) increased in both groups following γ‐T supplementation consistent with greater baseline FMD. Baseline nitro‐γ‐T and total cysteine and postprandial malondialdehyde AUC were lower in all men following γ‐T whereas myeloperoxidase and C‐reactive protein were unchanged. Thus, short‐term γ‐T supplementation likely improves VEF by decreasing oxidative and nitrative damage, but without affecting insulin resistance or inflammatory responses. Support provided by International Life Sciences Institute to RSB.

Grant Funding Source: International Life Sciences Institute
Maternal serum retinol and the outcome of pregnancy (Camden Study)
Authors: Scholl Theresa O , Chen Xinhua , Schluter Margaret , Leskiw Maria , Stein T. Peter
Published: June 2021

Vitamin A affects gene expression and thus influences diverse physiologic processes. Both low and high intakes of Vitamin A are teratogenic in animals. During pregnancy low maternal levels of serum retinol in malnourished populations, are associated with poor fetal growth, preterm delivery and infant mortality. The extent to which either an excess or a deficit of retinol is linked to adverse pregnancy outcomes in other populations is not known. We assayed maternal serum retinol at week 28 gestation by HPLC in 1180 gravidae from Camden, NJ. High levels (highest quintile >54.8ug/dl) correlated (p<0.05) with increased energy adjusted intakes of total vitamin A, beta carotene and carotene. High retinol levels were associated with significantly increased risks of small for gestation birth (Adjusted Odds Ratio (AOR) 1.88, 95% Confidence Interval (CI) 1.19–2.97), preterm delivery (AOR=1.87, 95% CI 1.56–3.02) and very preterm delivery (<32 weeks) AOR=4.15, 95% CI 1.41–12.22). Maternal serum retinol was higher (~ 30%, p=0.029) among gravidae (N=7) with live born neonates who died before day 28. Women in the lowest quintile, where serum retinol levels were low (<30ug/dl) but not suggestive of deficiency, did not show an increase in risk. Thus in populations that are not Vitamin A deficient, high levels of maternal serum retinol are associated with an increased risk of adverse pregnancy outcomes.
Provitamin A from Golden Rice is as effective as provitamin A in oil to children
Authors: Tang Guangwen , Yin Shi‐an , Wang Yin , Grusak Michael , Russell Robert
Published: June 2021

Spinach and pure β‐carotene (β‐C) in oil capsule are good sources for vitamin A. A new transgenic product, Golden Rice (GR), is enriched with β‐C genetically. To determine its vitamin A value as compared with spinach or pure 2H8 labeled β‐C (M+8), we conducted a study with healthy Chinese school children (age 7–8y). Intrinsically labeled GR and spinach were obtained by growing plants in a nutrient solution containing ~25% 2H2O. The GR β‐C and spinach β‐C were enriched with deuterium with the highest abundance at M+9. An oil capsule of 13C10 ‐ retinyl acetate (M+10) was taken as a reference dose to determine the β‐C to retinol conversion factors. Serum samples collected from the subjects were analyzed using GC/ECNCI‐MS for the enrichments of labeled retinol by monitoring the isotopes of M+4 (derived from 2H8 β‐C), M+5 (from GR or spinach β‐C) and M+10 (from 13C10 ‐ retinyl‐acetate). By using the response to the dose of 13C10 ‐ retinyl acetate (0.5 mg) as a reference, our preliminary results (using area under the curve of percent enrichment at days 1, 3, 7, 14, and 21 after the labeled doses) showed that the conversions of pure β‐C (0.5 mg), GR β‐C (0.6 mg), and spinach β‐C (1.4 mg) to retinol were 2.0, 2.1, and 7.3 to 1 by weight, respectively. These results indicate that GR can be as effective as β‐C in oil in contributing to the daily vitamin A requirements of children, thus, to improve population‐based vitamin A adequacy.
Analysis of mineral contents in bottled natural water and estimation of theirs intakes in Korean adults
Authors: Choi Mi‐Kyeong , Kim Moo‐Kyung , Sim Jin‐Ah , Eom Hee‐Jung , Kim Myung‐Hee , Kang Myung‐Hwa
Published: June 2021

Minerals play an important role in the body as essential nutrients. However, it's not easy to meet DRIs because food sources of minerals are limited. Recently, consumption of bottled natural water is increasing because of water pollution and distrust in tap water in Korea. The present study was conducted to investigate mineral contents in bottled natural water and theirs intakes in Korean adults. We analyzed 8 minerals in 7 kinds of bottled natural water by ICP‐spectrometer and conducted survey on intake status of water and natural water of 400 Korean young adults. The mean contents of Ca, Mg, Fe, Zn, Cu, Mn, Se, and Mo in bottled natural water were 22.5±22.5 mg/L, 10.6±10.0 mg/L, 0.3±0.2 ug/L, 2.1±1.5 ug/L, 1.2±0.1 ug/L, 1.4±0.4 ug/L, 1.9±1.0 ug/L, 3.3±0.8 ug/L. The mean age, height, weight, and BMI were 22.8 years, 174.9 cm, 68.6 kg, 22.4 kg/m2 for male (n=150) and 21.3 years, 162.0 cm, 51.1 kg, 19.5 kg/m2 for female (n=250). The daily intakes of total water and natural water as water itself were 670.3 mL and 212.2 mL for male and 488.0 mL and 132.7 mL for female. The daily intakes of Ca, Mg, Fe, Zn, Cu, Mn, Se, and Mo through natural water were 4.8 mg, 2.2 mg, 0.06 ug, 0.4 ug, 1.2 ug, 0.3 ug, 0.4 ug, and 0.7 ug for male and 3.0 mg, 1.4 mg, 0.04 ug, 0.3 ug, 0.7 ug, 0.2 ug, 0.3 ug, and 0.4 ug for female. To sum up these results, the contents of Ca, Mg, and Se in bottled natural water were relatively high and the daily intakes of these minerals were 0.5~1.0 % DRIs.
A Randomized Trial of Multivitamin Supplementation in Children with Tuberculosis in Tanzania
Authors: Mehta Saurabh , Mugusi Ferdinand M , Bosch Ronald J , Aboud Said , Chatterjee Anirban , Finkelstein Julia L , Fataki Maulidi , Kisenge Rodrick , Fawzi Wafaie W
Published: June 2021

Background

Multivitamin supplementation may enhance the health of children with tuberculosis (TB); however, its efficacy as an intervention has not been examined.

Methods

255 children with TB were randomized to receive either a daily multivitamin supplement or placebo in the first eight weeks of anti‐TB therapy. They were followed up for the duration of supplementation through clinic and home visits to assess anthropometric indices and laboratory parameters, including hemoglobin and albumin.

Results

Multivitamin supplementation resulted in a significant improvement in hemoglobin levels at the end of follow‐up; the median increase in children receiving multivitamins was 1.0 g/dL, compared to 0.4 g/dL in children who were given placebo (p<0.01). No significant differences were observed in growth indices, including height and weight gain. Among children who were also co‐infected with HIV (n=87; 34%), daily multivitamins had a smaller but statistically significant increase in albumin levels compared with placebo.

Conclusions

Multivitamin supplementation for eight weeks improved the hematological profile of children with TB. Larger studies with a longer period of supplementation are needed to confirm these findings and assess the effect of multivitamins on clinical outcomes including treatment success and growth failure.

ClinicalTrials.gov Identifier: NCT00145184

Grant Funding Source: NIH 3U01AI045441
Zinc absorption from traditional meals by vitamin A‐depleted Bangladeshi women is greater in those with altered intestinal permeability, while iron absorption is not affected
Authors: Perez‐Exposito Ana Beatriz , Woodhouse Leslie , Haskell Marjorie J , Jamil Kazi M , Barfod Gry , Wahed Mohammed A , Brown Kenneth H
Published: June 2021

Background

Zinc and iron deficiencies have been associated with altered intestinal mucosal function.

Objectives

To examine the relationship between intestinal permeability and zinc and iron absorption in non‐pregnant, vitamin A‐depleted Bangladeshi women (plasma retinol < 1.05 umol/L).

Methods

Zinc absorption from a traditional test meal was assessed using the dual stable isotope tracer ratio method, and iron absorption was assessed by erythrocyte incorporation of a stable iron isotope 10 days after the oral dose. Intestinal permeability was evaluated with the lactulose‐mannitol (L:M) test.

Results

Fractional zinc absorption (FZA) was greater in women with altered intestinal permeability (urinary L:M recovery ratio >0.07) than in women with L:M ≤0.07 [FZA = 36.1 ± 9.4 % vs. 24.9 ± 10.3 %, respectively, p = 0.038]. Iron absorption from meals did not differ by intestinal permeability status [2.9 % (1.4, 5.8) vs 2.6 % (1.8, 3.8) p = 0.79]. Zinc absorption was marginally correlated with the urinary lactulose:creatinine ratio (p = 0.07).

Conclusions

Zinc absorption from a traditional meal in vitamin A‐depleted Bangladeshi women is greater in those with altered intestinal permeability than in those with normal permeability, but iron absorption is not affected

Funding was provided by HarvestPlus. Ana Perez‐Exposito was partially funded by the International Nutrition Foundation/Ellison Medical Foundation.
Three models of HemoCue photometers: comparison of venous blood hemoglobin concentrations and influence of heat on cuvettes
Authors: Mandava Usha , Drammeh Bakary S , Zhang Mindy , Schleicher Rosemary L , Pfeiffer Christine M
Published: June 2021

Three models of portable HemoCue photometers are often used to measure hemoglobin (Hb) as part of nutrition surveys in developing countries: Hb‐B, Hb‐201 and Hb‐301. We investigated the comparability between and within models and simulated high temperature storage of cuvettes during field surveys. Adult venous blood Hb (n=35) was measured in duplicate using 2 photometers per model and optimally stored cuvettes. Hb (n=24) was also measured in duplicate using non‐optimally stored cuvettes (37°C for 6 hours, 1–3 days and 1–3 weeks). We calculated mean and 95% confidence intervals (CI) for Hb concentrations and mean changes from optimal for heated cuvettes, and performed ANOVA and t‐tests using SAS software. Mean Hb concentration (14.3 g/dL; 95% CI: 13.8–14.7 g/dL) was not significantly different (p=0.37) between and within models, thus the 3 HemoCue models are interchangeable. For cuvettes stored at 37°C for up to 3 weeks, in no case was the difference from optimal storage ≥ 1%. Extended exposure to heat adversely affected Hb‐201 cuvettes but the effect was minimal. Hb‐301 cuvettes have the widest allowable storage temperature range (10–40°C vs 15–30°C) and provided robust data for up to 3 weeks exposure to heat.
Bioconversion of B‐cryptoxanthin into polar metabolites other than vitamin A
Authors: Mein Jonathan Ryan , Dolnikowski Gregory , Russell Robert , Wang Xiang‐Dong
Published: June 2021

Several well‐implemented cohort studies have shown dietary intake or blood levels of β‐cryptoxanthin, but not other pro‐vitamin A carotenoids such as β‐carotene and α‐carotene, to be strongly associated with a reduced risk of lung cancer. This has raised an important question as to whether the potential protective activity of β‐cryptoxanthin, a mono‐hydroxy carotenoid, is due to the intact molecule or formation of unique metabolites. The objective of the present study was to investigate the excentric cleavage activity of mammalian carotene‐9′,10′‐oxygenase (CMO2) towards β‐cryptoxanthin. We found that baculovirus‐generated recombinant ferret CMO2 cleaved β‐cryptoxanthin at both the 9,10 and 9′10′ double bond in a protein‐, time‐ and dose‐dependent manner. In addition to the expected apo‐10′‐carotenal and β‐ionone cleavage products, HPLC and LC‐MS analysis identified the formation of two unique metabolites: 3‐OH‐apo‐10′‐carotenal and 3‐OH‐β‐ionone. Furthermore, in the presence of NAD+, incubation of 3‐OH‐apo‐10′‐carotenal with ferret liver S9 lysate resulted in the dose‐dependent formation of 3‐OH‐apo‐10′‐carotenoic acid. In conclusion, the bioconversion of β‐cryptoxanthin by CMO2 into 3‐OH‐apo‐10′‐carotenoids is significant because 3‐OH‐apo‐carotenoids may represent biologically important metabolites with specific biological activities related to human health.

Grant Funding Source: R01CA104932
Tomato carotenoid bioaccumulation and gene expression of carotenoid cleavage enzymes are altered by soy germ in male Copenhagen rat tissues
Authors: Zuniga Krystle Erin , Erdman John W
Published: June 2021

Epidemiological studies have supported the hypothesis that dietary intake of soy foods or tomato products is associated with a lower risk of prostate cancer. However, little is known about how simultaneous consumption may impact absorption, tissue distribution, and metabolism of tomato carotenoids. Diets containing 10% tomato powder (TP) (n=17), 2% soy germ (SG) (n=16), neither (n=17), or a combination (TP+SG) (n=13) were fed to 12 week old male Copenhagen rats for approximately 24 weeks. Bioaccumulation of tomato carotenoids were lower in various tissues of rats fed TP + SG compared to TP alone, especially in androgen sensitive tissues. Testes, seminal vesicles, and ventral prostate of the TP+SG group had lower levels of lycopene (p=0.001, p=0.04, and p=0.002, respectively) and β‐carotene (p=.003, p=.003, p=.001, respectively) than the TP group. Gene expression of carotenoid 15,15′‐monooxygenase (CMOI) and carotenoid 9′10′‐monooxygenase II (CMOII) were measured in the liver, duodenum mucosa, and prostate. Rats fed TP had significantly higher hepatic CMOI expression compared to the control group (p=.046), but expression was not significantly different from rats fed SG or TP+SG. Liver expression of CMOII was not significantly different between groups. Our results suggest that dietary soy germ may alter uptake and/or metabolism of carotenoids. Research supported in part by USDA NNF 2005‐03750.

Grant Funding Source: NIH
A lutein enriched diet modified lipoprotein subfractions and prevented cholesterol accumulation in aortas in guinea pigs
Authors: Kim Jung Eun , Leite Jose O , DeOgburn Ryan , Clark Richard M , Fernandez Maria Luz
Published: June 2021

Two groups of guinea pigs were fed a control diet (n =9) or a diet containing 0.1% lutein (n =10) for 12 wk to evaluate the effects of this carotenoid on lipoprotein metabolism and potential protection against atherosclerosis. Lipoprotein subfracion and size were measured using nuclear magnetic resonance, hepatic and aortic lipids were extracted with chloroform:methanol to measure tissue cholesterol. Lutein concentrations in liver were determined by HPLC. Plasma total and LDL cholesterol were not different between groups, however, the lutein treated group had higher concentrations of plasma total VLDL and LDL particles (P < 0.05) and lower concentrations of large HDL particles (P < 0.01) and higher concentrations of medium and small HDL compared to the control group. The increases in these particles may be related to the way lutein is transported in plasma, mainly in LDL and small HDL. In contrast, aortic cholesterol was lower in the lutein (3.7 ± 1.0 mg/g) compared to the control group (6.0 ± 0.9 mg/g). Guinea pigs from the lutein group had an average of 23.83 ± 5.61nmol/g of lutein while this carotenoid was not detected in the control group. Hepatic lutein concentrations were negatively correlated with small LDL (r = −0.561, P < 0.05) and with aortic cholesterol (r=−.734, p< 0.01). These results suggest that lutein in the liver is related to less formation of small LDL particles and therefore lower deposition of cholesterol in the aorta.
Aggregation Behavior of Xanthophylls
Authors: Landrum John T. , Babino Darwin
Published: June 2021

Aggregation of carotenoids in biological systems influences their properties. The objective of this study is to investigate the extent to which carotenoid aggregation varies as a function of the polarity of the solvent mixture and temperature. The visible spectra of lutein, zeaxanthin and the monomethyl ether of lutein were measured in alcohol/water mixtures that favor aggregation. Over a range of temperatures in these solvent mixtures the extent of aggregation changes, in some cases dramatically. Using a two‐component, monomer/aggregate model an estimate of the enthalpy for the aggregation by these xanthophylls has been made. Our results for these systems provide insight into the energetics of xanthophyll aggregates. Lutein and zeaxanthin exhibit little difference at low concentration in their aggregation properties. Measured ΔH (kcal/mole) values for aggregation range between +5.9 and +11.5 for lutein and +3.9 and +10.3 for zeaxanthin and are sensitive to solvent composition. Methylation of the 3′‐OH group of lutein disrupts aggregate formation showing that H‐bonding at both ends of the carotenoid is a significant contributor to the stability of aggregates.
Intracellular accumulation and metabolism of beta‐carotene and lycopene in normal and non‐small cell lung cancer cells
Authors: Marisiddaiah Raju , Gong Xiaoming , Wiener Doris , Rubin Lewis P.
Published: June 2021

β‐Carotene (BC), lycopene (lyc) or total dietary carotenoids have protective role in non‐small cell lung cancer (NSCLC). However, high‐dose BC increased NSCLC incidence in high‐risk individuals. Limited knowledge of uptake and metabolism of BC and lyc in human lung cells prompted this investigation in immortalized human bronchial epithelial (BEAS‐2B) and alveolar epithelial‐derived adenocarcinoma (A549) cells. Our results show that BEAS‐2B and A549 cells respectively express high levels of CMO2 and CMO1 mRNA (qRT‐PCR) and protein (western blot). We analyzed cellular BC, lyc and putative metabolites by HPLC after 3, 6, 24 and 48h exposures to 3 and 5μM of BC or lyc. BC and lyc were taken up in a time‐ and dose‐dependent manner. Incubation with 3 and 5μM BC resulted in 306 and 374 pmol BC (A549), and 512 and 306 pmol BC (BEAS‐2B) per million cells. Interestingly, BC was metabolized to retinal in A549 cells after 6h, consistent with the high level of CMO1 expression. A reduced retinal accumulation observed after 48h appears to be due to further retinoid metabolism. In contrast, BEAS‐2B cells showed no detectable BC metabolites, indicating BC is not significantly metabolized by these cells. Lyc treatment generated several isomers and oxidized metabolites in both cell lines. After 24h treatment with 3 and 5μM lyc, accumulation was 93 and 102 pmol lyc (A549), and 491 and 614 pmol lyc (BEAS‐2B) per million cells. We conclude that normal and NSCLC cells may differently metabolize BC and lyc. These findings, and our other data, suggest that a shift in carotene metabolism (a) accompanies NSCLC carcinogenesis and (b) may underlie the deleterious effects of high‐dose BC in lung cancer clinical trials. Further studies are needed to characterize metabolism in normal and abnormal lung epithelium.

Grant Funding Source: NIH HD42174
Efficiency of intestinal absorption of beta‐carotene (BC) is not correlated with cholesterol (CHL) absorption in humans
Authors: Fleshman Matthew Kintz , Cope Keary A , Novotny Janet A , Riedl Ken , Schwartz Steven J , Jones Peter J , Baer David J , Harrison Earl H
Published: June 2021

The intestinal absorption of BC and of CHL varies among individuals. Experiments in vitro and in mouse models suggested that BC and CHL shared certain molecular mechanisms of absorption. A study was conducted to investigate the variability in BC absorption and its conversion to vitamin A and to compare the efficiency of absorption of BC with that of CHL. Ten men consumed a 5 mg dose of deuterium labeled BC (BCd8), with 6 subjects repeating the dose 2 months later. Blood was collected hourly for 9 h, and enrichment of chylomicrons with BCd8 and retinyl‐d4 esters (REd4) was determined by HPLC‐MS. CHL absorption was measured in the same subjects by the oral/IV dual isotope ratio method using deuterated CHL and 13C CHL and ranged from 28% to 60%. Chylomicron BCd8 AUC ranged from 0.047 to 0.414 μM‐hr and the CM REd4 AUC ranged from 0.102 to 2.59 μM‐hr. For subjects who repeated the treatment, the total BC absorbed at the two times was correlated (r=0.99; p<0.0001). The conversion of BCd8 to REd4 ranged from 37–77%, and the extent of conversion at the two times was correlated (r=0.77; p=0.08). There is marked inter‐individual variability in BC absorption and conversion to RE. In contrast, intra‐individual variability in BC absorption and its conversion to RE is low. Importantly, there was no correlation between an individual's efficiency of absorption of BC and their efficiency of absorption of CHL (r=0.21; p=0.56). HL049879 & USDA/ARS
Carotenoid levels in ageing human brain with reduced cognitive function
Authors: Ferruzzi Mario G. , Ho Lap , Pasinetti Giulio M.
Published: June 2021

While increased plasma carotenoid levels have been associated with protection of cognitive function in humans, only limited information is available on carotenoid levels in human brain tissues through ageing. To better understand if brain carotenoid profiles change as a result of enhanced aging and declined cognitive function, human brain tissues (frontal cortex) were collected and analyzed post‐mortem from young‐old (YO; n=15) (<85 years) and old‐old (OO; n=15) (>85 years) individuals defined with normal cognitive function (NCF), mild (MCI) or severe cognitive impairment (SCI). Lutein, zeaxanthin and β‐cryptoxanthin were predominant carotenoid species in brain tissues from both normal and cognitively impaired accounting for >70% of total carotenoid levels in brain tissues. Higher total carotenoid levels were observed in tissues from OO (347.1 pmol/g) compared to YO (147.8 pmol/g) subjects with normal cognitive function. Additionally, a marked decrease was observed in total carotenoid content of OO tissues from individuals with mild cognitive impairment relative to normal function (224.6 versus 347.1 pmol/g respectively). Carotenoid levels in both OO and YO brain tissues with severe cognitive impairment did not different from normal individuals. These data suggest that carotenoid levels in brain tissue may correlate with ageing and may decline with mild cognitive impairment. The clinical significance of these results and the extent to which these differences are associated with specific markers of cognitive function merits future consideration.
Production of [13C]‐lycopene from high lycopene tomato cell suspension cultures
Authors: Engelmann Nancy J. , Rogers Randy B. , Rupassara S. Indu , Garlick Peter J. , Lila Mary Ann , Erdman John W.
Published: June 2021

The goal of this work was to use tomato cell suspension cultures treated with enzyme inhibitors and grown with [13C6]‐glucose as a carbon source to produce labeled carotenoids (CAR) for human metabolism research. To promote CAR accumulation, the high lycopene (LYC) tomato cell suspension line ‘Ailsa Craig’ hp‐1 was grown with carotenogenic enzyme inhibitors norflurazon (NORF), 2‐(4‐chlorophenyl‐thio) triethylamine (CPTA), both herbicides (BOTH), or neither (CONT). Significantly greater combined CAR [phytoene (PE), phytofluene (PF), and LYC] yields were obtained with the NORF, CPTA, and BOTH treatments (5.2 ± 0.7, 3.6 ± 0.4, 4.2 ± 0.4 mg/L, respectively) than CONT (0.3 ± 0.1 mg/L). Different PE, PF, and LYC profiles were obtained where CPTA led to 0.26 ± 0.02, 0.14 ± 0.02, and 3.18 ± 0.45 mg/L; NORF led to 4.96 ± 0.68, 0.20 ± 0.06, and 0.06 ± 0.02 mg/L; BOTH led to 2.98 ± 0.14, 0.18 ± 0.05, and 1.05 ± 0.36 mg/L; CONT led to 0.18 ± 0.09, 0.02 ± 0.00, and 0.06 ± 0.01 mg/L, respectively. To biolabel LYC the hp‐1 cell line was grown in medium containing 30 g/L [13C6]‐glucose and CPTA. LYC, PE, and PF yields from labeled cultures were 2.5 ± 0.1, 0.3 ± 0.0, 0.1 ± 0.0 mg/L, respectively. Mass isotopomer distribution analysis for LYC [13C]‐enrichment from a pilot labeling run was performed using LC‐MS/MS, and 82% of the LYC molecules had at least 34 of 40 carbons labeled, and the greatest percent (17%) was uniformly labeled. (NIH/NCI CA 112649‐01A1)
Quercetin causes dose‐related increases in energy expenditure and PGC1α in muscle of C57BL/6J mice
Authors: Henagan Tara Michelle , Lenard Natalie R , Rietschier Helena , Anaya Stephanie , Gettys Thomas W , Cefalu William T , Stewart Laura K
Published: June 2021

We explored the protective effect of varying doses of the antioxidant quercetin on mitochondrial function in C57BL/6J mice on a high fat (HF) diet. Six week old male C57BL/6J mice (N = 192) were randomly assigned to one of the following diets: HF (45% kcal fat), HF + 600ug quercetin/day (600Q), HF + 250ug quercetin/day (250Q) or HF + 50ug quercetin/day (50Q). Body weight (BW), body composition (BC), and food consumption (FC) were measured weekly. At 3 weeks (wk) and 8 wk, energy expenditure (EE), respiratory exchange ratio (RER), and activity were assessed. At the 3 and 8wk time points, animals were sacrificed and quadriceps muscle was used for quantification of peroxisome proliferator‐activated receptor gamma coactivator‐1 alpha (PGC1α) mRNA by real time PCR. The assessed variables did not differ at 3 wk, but at 8 wk 50Q significantly increased EE (P < 0.05) but did not change PGC1α Conversely, the 600Q group exhibited a decrease in EE yet higher levels of PGC1α in comparison to HF (P < 0.05). 250Q had no difference in EE at 8 wk but increased PGC1α (P < 0.05). There were no differences in BW, BC, or FC among groups at 8wk. We conclude that 8wk of quercetin supplementation causes dose‐related metabolic responses that may be indicative of protective mitochondrial adaptations to a HF diet, independent of a reduction in BW or a change in BC.

Grant Funding Source: Botanical Research Pilot Funding Program ‐ Pennington Biomedical Research Center