bluuebunny/crossref_metadata_2025 huggingface dataset:
FDA's Evaluations of GRAS Food Ingredients
Authors: Dausch Judith
Published: June 2021

Nutritional substances have been the subject of numerous Generally Recognized as Safe (GRAS) notices that have been voluntarily submitted to FDA via its GRAS Notification Program. FDA's evaluations of manufacturers' GRAS notices depends on the regulatory framework for food ingredients laid out under the Federal Food, Drug, and Cosmetic Act (the Act). Under the Act, there is a single standard for “safe” pertaining to food additive uses and GRAS ingredient uses. The regulatory framework for ingredients added to food, the safety standard, and the criteria that sets GRAS ingredient uses apart from food additive uses, namely, availability of appropriate safety data to qualified experts and consensus among those experts, will be the focus of this presentation. Case studies highlighting the types of data and information in GRAS notices for nutritional substances, such as fiber, and how FDA evaluated such notices will be used to reinforce key points.
Evaluative Ethnography is useful for Understanding the Impact Pathways of Performance‐Based Incentives on Health Worker Motivation
Authors: Schuster Roseanne , de Sousa Octávio , Reme Anne‐Kathe , Pinault Delphine , Young Sera
Published: June 2021

Performance‐based incentives (PBIs) have shown potential to improve the delivery of health and nutrition services to vulnerable populations. Strikingly, assessments of PBIs rarely incorporate qualitative approaches, making it difficult for policymakers to fully evaluate their utility. We employed evaluative ethnography to assess the impact of a longitudinal‐control PBI intervention to improve prevention of vertical transmission of HIV service delivery in rural Mozambique.

Evaluative ethnography is a structured and theory‐grounded approach to qualitative research. The motivation, opportunity, and ability framework and focused ethnographic methods characterized our evaluative ethnographic approach.

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Table 1. Utilization of focused ethnography and quantitative methods to assess impact of the performance‐based incentive intervention on delivery of prevention of vertical transmission of HIV (PVT) services

Method
Participants
Intervention Phase
n

Evaluative Ethnography Methodology

Semi‐structured interviews
Women accessing PVT services, their family members, community leaders, facility‐ and community‐based health workers
Formative
67

Group discussions
Facility‐ and community‐based health workers, district leaders
Formative, on‐going implementation
100

Participant observation
Facility‐ and community‐based health workers
On‐going implementation
140

Semi‐structured interviews
Facility‐ and community‐based health workers, district leaders
Final evaluation
15

Quantitative Assessment Methodology

Surveys of health worker motivation
Facility‐ and community‐based health workers
Formative, on going implementation, final evaluation
147

Participant observation and exit interviews revealed the major barriers to the intervention (e.g. obstructionist officials, resource constraints). This was critical to understanding the quantitative finding that the PBIs did not significantly affect health worker motivation or delivery of services within the 12‐month implementation period.

Although PBIs did not impact service delivery, evaluative ethnography allowed us to identify the principal problem areas. Considering the challenging nature of nutrition and health interventions in low‐resource health systems, we demonstrate that evaluative ethnography can augment conventional quantitative assessments.
Nonessential Amino Acids as Nitrogen Sources in Adult Men Examined Using the Indicator Amino Acid Oxidation Technique
Authors: Cooper Leah , Ball Ron , Pencharz Paul , Sakai Ryosei , Elango Rajavel
Published: June 2021

Nutritionally, there is a dietary requirement for the essential amino acids (EAA) but also a requirement for nitrogen (N) intake for the de novo synthesis of the nonessential amino acids (NEAA). It has been suggested that some NEAA may be more metabolically important than others. The goal of this study was to determine whether among the NEAA (Ala, Arg, Asn, Asp, Gln, Glu, Gly, Pro, Ser) there was an ideal N source using the indicator amino acid oxidation (IAAO) technique.

Subjects were maintained on an adaptation diet for 2 days prior to each test day. Each subject participated in 10 test diet intakes, assigned randomly. One test intake was a base diet consisting of only the EAA provided at the recommended dietary allowance. All other test intakes involved the base diet with the addition of one NEAA to meet a 50:50 ratio of EAA: NEAA on a N basis. The diets were isocaloric and isonitrogenous, given as hourly meals. Each study day followed the IAAO protocol using 13C‐Phenylalanine as the indicator. Breath and urine samples were collected at baseline and isotopic steady state. Enrichments of 13C in breath were analyzed by continuous‐flow isotope ratio mass spectrometry to calculate F13CO2.

Repeated measures ANOVA with post hoc multiple comparisons were done on the F13CO2 data. Preliminary results indicate that 7 of the 9 NEAA decrease IAAO significantly and suggest that in healthy males, most NEAA are good N sources, in the presence of adequate EAAs.

(Support by Ajinomoto Co. Inc.)
Branched‐chain Amino Acid Catabolism is Required for Muscle Cell Differentiation
Authors: Dhanani Zameer , Dhaliwal Kanwar , Adegoke Olasunkanmi
Published: June 2021

The importance of branched‐chain amino acids (BCAAs) in promoting skeletal muscle anabolism has been well studied. BCAAs isoleucine, leucine, and valine have been shown to have a profound effect on activating anabolic signaling pathways in skeletal muscle. This occurs in part by the upregulation in activity of the mammalian target of rapamycin complex‐1 (mTORC1), resulting in increased protein synthesis. However, the regulation of branched‐chain amino acids during the development of muscle remains yet to be elucidated. Here, we studied BCAA metabolism during a 5‐day differentiation of L6 myoblasts. Although no change in intracellular BCAA concentrations was observed during differentiation, L6 cells cultured in the absence of leucine were severely impaired in their ability to differentiate as expression of myosin heavy chain (MHC) was completed abrogated at day 5. Two enzymes which are critical for BCAA metabolism are the branched‐chain amino transferase‐2 enzyme (BCAT2) and the branched chain α‐keto acid dehydrogenase complex (BCKD). BCAT2 catabolizes BCAAs to their corresponding alpha‐keto acids, which are then irreversibly decarboxylated by the BCKD complex. The abundance of BCAT2 did not change during differentiation, whereas levels of the E1α subunit of BCKD was increased 7x on day 5 compared to day 1 (p<0.05). Finally, when either BCAT2 or BCKDE1α was knocked down in the presence of leucine, myoblast differentiation was abrogated (BCKDE1α RNAi: MHC decreased 4.5x from day 3 to day 5 (p<0.05); BCAT2 RNAi: MHC levels completely abolished). Our findings suggest that BCAA catabolism may be a critical process in facilitating muscle differentiation.
Enteral Supplementation with β‐Hydroxy‐β‐Methylbutyrate Increases Muscle Protein Synthesis in Neonatal Pigs
Authors: Kao Michelle , Columbus Daniel , Suryawan Agus , Steinhoff‐Wagner Julia , Hernandez‐Garcia Adriana , Nguyen Hanh , Davis Steven , Fiorotto Marta , Davis Teresa
Published: June 2021

Premature infants are at risk of poor growth due to an inability to achieve adequate protein intake. Recently, we showed that acute parenteral administration of the leucine metabolite, β‐hydroxy‐β‐methylbutyrate (HMB), enhances protein synthesis in skeletal muscle of neonates by stimulating mTOR‐dependent translation initiation. The purpose of this study was to determine the effects of enteral administration of HMB on protein synthesis in skeletal muscle of the neonate and to identify the mechanisms involved. Overnight fasted neonatal piglets were either studied immediately (Fasted) or fed for 24 h by intermittent bolus 1 of 5 diets: low protein diet (LP; 8.3 g protein•kg‐1•d‐1), high protein diet (HP; 18 g protein•kg‐1•d‐1), or LP diet supplemented with 4 (HMB 4), 40 (HMB 40), or 80 (HMB 80) µmol HMB•kg‐1•d‐1. Maximum plasma HMB concentrations were 10, 45, and 80 nmol•ml‐1 in HMB 4, 40, and 80, respectively, 5 nmol•ml‐1 in both LP and HP, and 3 nmol•ml‐1 in Fasted piglets. Compared to Fasted piglets, HMB 40 and HMB 80 increased (P < 0.05) intracellular concentration of HMB in longissimus dorsi. Muscle protein synthesis increased with HMB 80 compared to LP, HMB 4, and Fasted (P < 0.05), and increased further with HP (P < 0.05). Phosphorylation of both S6K1 and 4EBP1 and formation of the active eIF4G‐eIF4E complex increased (P < 0.05) with HMB 80 compared to LP. Our results suggest that enteral HMB supplementation increases protein synthesis in skeletal muscle of neonates with limited protein intake through enhanced activation of translation initiation.
Effect of different types of dietary proteins on liver and muscle mTORC1 pathway after nutritional recovery
Authors: Rodriguez‐Gaytran Cynthia , Márquez‐Mota Claudia , Tovar Armando , Torres Nimbe
Published: June 2021

It is known that protein synthesis is regulated by mTORC1 (mechanistic target of rapamycin). When activated, mTORC1 promotes protein synthesis mainly by phosphorylation of S6K1 and 4E‐BP1. There is an interest to study some dietary proteins or dietary protein combinations for their possible use in the nutritional recovery. In the present work we studied the effect of different types and combination of dietary proteins on the mTORC1 pathway after a period of undernutrition. Sprague Dawley rats (n= 18 per group), were given a diet with a low concentration of casein (0.5% ) during 21 days in a restricted feeding period (9 hrs). After that period, rats were divided in 7 groups and were fed with one the following diets containing different protein sources: 1) 0.5% casein, 2) 20% casein, 3) 20% soy protein, 4) 20% black bean protein, 5) 20% corn protein, 6) 10% black bean‐10% corn proteins, 7) 10% soy protein‐10% corn proteins. Rats were fed with the corresponding diets during 1, 7, or 14 days. Our results showed that rats fed with casein, soy protein, soy protein‐corn or black bean‐corn had similar weight gain and recovery, meanwhile rats fed with 0.5% casein showed the lowest weight gain followed by 20% corn protein. The phosphorylation of liver and muscle mTORC1 was similar among the groups except for the groups 1 and 5. Rats fed with the combination of dietary proteins or 20% casein showed an increase in pS6K in comparison with rats fed 0.5% casein or 20% corn protein. Thus, the results indicate that the combination of dietary proteins with corn protein probably enhance protein synthesis in a similar extent than 20% casein due to the high leucine content in the corn protein through the activation of mTOR signaling pathway via S6K.
Lipid Emulsion Halts the Progression of Non‐Alcoholic Fatty Liver Disease (NAFLD) in Nonobese Mice Fed a Liquid High Carbohydrate Diet
Authors: Huang Kuan‐Hsun , Hao Lei , Smith Philip , Patterson Andrew , Ross A Cathatine
Published: June 2021

A two‐hit hypothesis has been proposed for the etiology of NAFLD: first, increased lipogenesis results in triacylglycerol (TG) accumulation, then inflammatory stimuli, e.g., saturated fatty acids (SFA)‐induced lipotoxicity and inflammation, cause disease progression. Previously, we showed that NAFLD‐like symptoms developed in mice fed a liquid high carbohydrate diet (76% en‐%, HCD) for 5‐wk, while mice fed HCD containing 13.5% (en‐%) of lipid emulsion (LE, Intralipid®) exhibited less hepatic TG accumulation and inflammation. However, it is unknown whether LE can cease the first hit of NAFLD once TG accumulation has begun. To test this, male C57BL/6 mice (6 wk; n=8/group) were fed either HCD for 5 wk (HCD) or HCD for 2.5 wk followed by HCD with LE for 2.5 wk (HCD/LE). At this time, liver TG in mice fed HCD/LE was 60% lower (P<0.001) as compared to mice fed HCD. Hepatic mRNAs of PNPLA3, associated with NAFLD,and several lipogenic genes, e.g., acetyl‐CoA carboxylase 1, fatty acid synthase, and stearoyl‐CoA desaturase 1, assayed by qRT‐PCR, were decreased by 42‐93% in HCD/LE vs. HCD mice (all P<0.001). We further analyzed liver total fatty acid composition, determined as fatty acid methyl esters by GC‐MS, and found that HCD/LE mice had lower levels of C14:0, C16:0 and C18:1 (n‐9) as compared to HCD mice (all P<0.001). In conclusion, in the presence of a HCD, LE can halt the first hit of NAFLD and lower the levels of saturated FA, reducing lipotoxicity and lipogenesis.
Role of the Lipolysis Stimulated Lipoprotein Receptor in Hepatic Lipid Homeostasis during the Postprandial Phase
Authors: Yen Frances , Akbar Samina , Layeghkhavidaki Hamed , Pinçon Anthony , Lanhers Marie‐Claire , Oster Thierry , Gregory Pauron Lynn , Corbier Catherine
Published: June 2021

Elevated postprandial lipemia (PPL) as an important risk factor for cardiovascular disease has been attributed to the accumulation of atherogenic remnant lipoproteins in the circulation. The hepatic lipolysis stimulated lipoprotein receptor, LSR, participates in the removal of these particles in the liver during the postprandial phase. LSR expression is regulated by leptin, and is reduced in obese mouse models which also display high PPL, suggesting a role of LSR in the regulation of lipid homeostasis. To determine the effect of reduced LSR expression on hepatic lipid metabolism, pathway‐specific qPCR arrays were used to determine mRNA expression profiles of genes related to lipid metabolism in livers from female 6‐month old LSR+/‐ mice and LSR+/+ littermates after 6 weeks on a standard (STD) or high‐fat diet (60 % kcal, HFD). Of the 84 genes studied, 32 were significantly down‐regulated in STD‐LSR+/‐ mice as compared to STD‐LSR+/+, a majority of which were involved in insulin and adipokine‐signaling pathways and were PPAR‐a target genes. Of these genes, 70% were also modified in HFD‐LSR+/+, suggesting that STD‐LSR+/‐ demonstrated a propensity towards a “high‐fat”‐like profile. Interestingly, the expression of 3 genes related to the inflammatory pathway: Ilβ, TNF‐α, and Il10 was increased in HFD‐LSR+/‐ mice, which could contribute to PPL‐associated inflammation. Our results show similar profiles of genes affected by either LSR heterozygosity or by HFD, pointing towards a significant contribution of LSR in the regulation of lipid homeostasis during the postprandial phase.

Grant source: French Ministry MESR, Région Lorraine
Effects of Differently Processed Soy Protein Compared to Milk Protein on Gut Microbial Composition: Possible Mechanisms for Cardiometabolic Effects of Soy
Authors: Butteiger Dustie , Hibberd Ashley , Napawan Nida , McGraw Nancy , Hall‐Porter Janine , Krul Elaine
Published: June 2021

Diet is a major factor influencing the composition of the gut microbiota, but studies investigating the effects of different proteins to modulate the microbiome are lacking. Soy protein consumption has several cardiometabolic benefits; however, the exact mechanism(s) whereby soy protein lowers serum lipids is unknown. This study is the first to investigate the effects of different protein sources on gut microbiota composition in a model of human lipid metabolism and to assess whether gut microbiota changes can account for the lipid lowering activity of soy protein. Golden Syrian hamsters (N=8/group) were fed a control semi‐purified diet containing 20% (wt) milk protein isolate (MPI) for 3 weeks, and then randomized to one of 4 treatment groups (MPI (control), soy protein concentrate 1 (SPC 1) or soy protein isolate (SPI 1) or (SPI 2)) for 6 weeks. The diets differed only in protein source. Feces were collected on day ‐1 and weeks 1, 3 and 6. Terminal blood samples were analyzed for serum lipids and intestinal sections were collected for 16S rDNA based microbial profiling. VLDL1+2 and non‐HDL‐C were significantly lower in all soy groups compared to the milk group suggesting that soy protein has its major effect to reduce hepatic lipogenesis. Total and LDL‐C were significantly lower in the SPC 1, and triglycerides were lower in the SPI 1 group vs the milk group respectively (soy groups NS diff from each other). There was greater microbial diversity with observed shifts in composition in hamsters fed soy diets vs MPI. Soy protein results in a different microbiota composition vs milk protein which may contribute to its lipid lowering effect. DuPont Nutrition & Health funded research.
Caspase1 Has Transcriptional Regulatory Effects Independent from That Mediated by IL1β and IL18 – Our Microarray and Meta‐analysis of Six Other Microarray Datasets
Authors: LI YAFENG , Yin Ying , Wang Hong , Yang Xiao‐feng
Published: June 2021

Caspase1 (Casp1) has been most intensely studied about its responsibility for proteolytic maturation of the pro‐inflammatory cytokines proIL1β and proIL18 into their mature and active cytokine forms. Recent studies have strongly indicated that Casp1 plays pivotal roles in regulation of metabolism, obesity, insulin resistance and cardiovascular disease, which are broader than the roles of IL1β and IL18. However, the underlying mechanisms remain unknown. We hypothesized that Casp1 has transcriptional regulatory effects independent from IL1β and IL18. To test this hypothesis, we performed RNA extraction from aortas of atherogenic ApoE‐/‐ mice and ApoE‐/‐/Casp1‐/‐ mice followed by the Affymatrix microarray analysis. We analyzed our Casp1 microarray data, used a novel method of meta‐analysis to analyze all gene expression datasets about Casp1, IL1b and IL18, which were uploaded by other teams into the Gene Expression Omnibus (GEO). The results include: 1) Casp1 has tissue‐specific regulatory effects on gene transcription globally and regulates 75 significantly among 7 different tissues; 2) then we compared the fold changes of each gene to that in 3 IL1β datasets. We found that Casp1 transcriptional regulatory effects are independent from that of IL1β; 3) similarly Casp1 transcriptional regulatory effects are also independent from that of IL18. In conclusion, our results have demonstrated for the first time that Casp1 has transcriptional regulatory effects which are independent from that mediated by IL1β and IL18.
Investigating the flap dynamics of HIV‐1 Protease
Authors: Stevens David , Miller Bill , Stansbury Rebecca , Springsted Gregory , Phelan Connor , Parish Carol
Published: June 2021

The potency of the HIV virus arises from the unknowing spread of the disease. Within the United States, it is estimated that one in four people newly infected with HIV is between the ages of thirteen and twenty‐four. (1) The HIV protease is an essential enzyme that cleaves three previral polyproteins leading to the maturation of the virus. The HIV virus mutates at a rapid pace, resulting in the emergence of HIV resistance to protease inhibitors. In 2003, it was suggested by Wu et al. that the flap tips of the HIV protease control enzyme activity. (2) Although the HIV‐1 protease flap dynamics have been studied extensively using both experimental and computational methods, rarely have all four flap tip conformations been observed in a single computer simulation. In addition, it has been suggested that one of the observed conformations was an artifact of crystal packing forces. (3) By performing long‐time scale simulations with two different force fields (CHARMM27 and AMBERff99SB) we were able to observe the existence of all four conformational states: closed, semi‐open, open, and wide‐open states. The lifetimes and percent occupancies of the conformations were determined through energy and structural analyses. Further analyses were performed to quantify stability and sampling of all simulations. These results will be presented and comparisons made with experimental results.

(1) AIDS.gov 2013.

(2) Wu, T. D.; Schiffer, C. A.; Gonzales, M. J.; Taylor, J.; Kantor, R.; Chou, S.; Israelski, D.; Zolopa, A. R.; Fessel, W. J.; Shafer, R. W. J Virol 2003, 77, 4836.

(3) Layten, M.; Hornak, V.; Simmerling, C. J. Am. Chem. Soc. 2006, 128, 13360.
Biophysical Characterization of Metal‐Deficient SOD5
Authors: Waninger‐Saroni Jessica , Gleason Julie , Taylor Alexander , Holloway Stephen , Culotta Valeria , Hart John , Galaleldeen Ahmad
Published: June 2021

Superoxide dismutase 1 (SOD1) is a homodimeric copper‐zinc containing enzyme that protects cells against oxidative damage through the rapid neutralization of reactive oxygen species (ROS). Its catalytic activity is dependent on the presence of a copper cofactor while zinc is important for its structural stability. Each subunit exhibits two extended loops, the zinc loop and the electrostatic loop, which both become disordered in the absence of zinc. SOD5 is a recently discovered extracellular copper‐only SOD found in the opportunistic pathogenic fungus Candida albicans that enables it to evade the host's immune response. SOD5 lacks the electrostatic loop but retains an ordered zinc loop even in the absence of zinc. In this study we determined the crystal structure of metal‐deficient SOD5. The protein was overexpressed in E. coli and found to be insoluble, necessitating denaturation, refolding, and further purification. SOD5 was stripped of metals, analyzed using sedimentation velocity experiments, concentrated, and crystallized. The crystal structure was determined by molecular replacement using SOD5‐Cu as a search model. Here we report that metal occupancy does not affect the oligomerization state of SOD5 and does not alter the tertiary structure of the molecule.
Regional Variation of Calcium Transients within Neonatal Cardiomyocyte
Authors: alvarez carlos
Published: June 2021

Cardiomyocyte contraction is preceded by a calcium transient, an influx of calcium into the cytosol causing a contraction to occur followed by relaxation which occurs as the intracellular calcium returns to resting level.We are investigating how calcium transient signals from the nuclear, perinuclear and more distant cytosolic regions of neonatal cardiomyocytes differ. Calcium transients exhibit significant differences in the rise and decay time courses between regions close to and relatively distant from the nucleus. Nuclear calcium transients are slower to occur and have broader peaks than cytosolic transients more distant from the nucleus. The distribution of SR around nuclear and perinuclear regions influences the behavior of calcium transient signals. We targeted four regions of interest (ROI) in the cell: (1) ROI 1 is the region with the least amount of nuclear calcium contribution; (2) ROI 2 refers to regions adjacent the nucleus where there is significant nuclear calcium contribution; (3) ROI 3 is within the nucleus; (4) ROI 4 refers to the signal averaged over the entire cell. In order to accurately define these four regions of interest the cells were stained with ER‐TrackerTM, a dye that binds to the SR, and with Fluo‐3. Calcium transients were recorded using Leica SP2 Fluorescent Laser Scanning Confocal Microscope. XT Scans were used to record all three regions of interest simultaneously for 30 sec and to compare these transients from an averaged signal recorded from the entire cell when cells were paced at 0.3 Hz. Neonatal ventricular myocyte measurements were obtained in an in‐line scan mode at 200 lines per second. Data files were exported in text format and graphed as bar charts and scatter plots. An averaged time to peak and averaged time to 50 percent signal decay were used to compare signal behavior between regions.
Doxorubicin‐coupled MNPs‐Fe3O4 with Electromagnetic Radiation Improve the Effectiveness of Doxorubicin on the T47D Human Ductal Breast Tumor Cell Line
Authors: Alobaid Sarah , Sinha Saion , Rossi Michael
Published: June 2021

Chemotherapeutic and anticancer drugs are limited by their serious side effects. Doxorubicin (DOX) is an anticancer compound that is used to treat breast cancer, but it can lead to irreversible and harmful side effects. Therefore, finding more effective and specific delivery methods may enhance the efficacy of DOX and decrease its side effects. Magnetic iron oxide (Fe3O4) particles (160 nm) were coupled with DOX and magnetically delivered to T47D breast cancer cells. The aim of this study is to develop a drug delivery system and to examine the effects of using magnetic nanoparticles (MNPs) on T47D breast cancer cells in vitro. T47D cells were treated with DOX, magnetic nanoparticles (MNPs), and with the DOX‐coupled magnetic nanoparticles complexes (DOX‐MNP) in present and absent of magnetic fields. Results showed that the cytotoxicity of DOX alone against T47D breast cancer cells was similar to the cytotoxic of DOX‐MNP after two days of treatment. However, assessing the cytotoxicity over longer time periods found that DOX‐MNP nanoparticles resulted in a greater decrease in cell viability compared to DOX treatment alone. The largest difference was seen after the five and six days of treatment. Cellular uptake of the complexes DOX‐MNP was increased by increasing the exposure time to the magnetic field. This was mirrored by an increase in the DOX‐MNP induced cytotoxicity. These results suggest that the use of chemotherapeutic compounds coupled to MNP may be an effective mechanism for their delivery to tumor cells allowing greater cytotoxicity at potentially lower doses.
A Trojan Horse for Light‐Triggered Delivery of Toxic Iron to Cancer Cells
Authors: Boice Emily , Kurtz Donald
Published: June 2021

Targeted exogenous agents that rapidly elevate reactive oxygen species (ROS) above toxic threshold levels has been touted as a potentially effective cancer therapeutic strategy. The “traditional” ROS‐mediated cancer therapy, referred to as photodynamic therapy (PDT), relies on dye‐photosensitized generation of singlet oxygen. However, therapeutically effective singlet oxygen generation requires well‐oxygenated tissues, whereas many tumor environments tend to be hypoxic. We describe here a potential alternative to PDT that would generate hydroxyl radical rather than singlet oxygen as the ROS, and do so in a tumor‐targeted fashion. Hydroxyl radical (OH•) is typically regarded as the most toxic among ROS, and can be generated by the Fenton reaction of ferrous iron with relatively low levels of hydrogen peroxide. We provide proof‐of‐principle for a strategy to flood tumor cells with Fenton reactive iron. The iron storage protein bacterioferritin (Bfr) has the unique property of binding 12 heme groups in its protein shell. We substituted the hemes with the well‐known photosensitizer, zinc protoporphryin IX and loaded the inner cavity of Bfr with ~1000 irons as a ferric oxyhydroxide polymer. We also fused the RGD4C tumor‐targeting peptide to the outer surface of the 24 subunits comprising the Bfr protein shell. Irradiation with visible light released ferrous iron and led to light‐dependent killing of C32 melanoma cells. Iron is a relatively cheap pro‐drug and our approach should be generally applicable to many types of cancers.
Antibody Directed Enzyme Prodrug Therapy: Discovery of novel genes, isolation of novel gene variants and production of long acting drugs for efficient cancer treatment
Authors: Goda Sayed
Published: June 2021

Back Ground

Antibody‐Directed Enzyme Prodrug Therapy (ADEPT) is a novel strategy to improve the selectivity of cancer treatment. The ADEPT uses the bacterial enzyme, glucarpidase to produce the antibody‐enzyme complex.

Repeated cycles of ADEPT and the use of wild type glucarpidase in detoxification are essential but are hampered by the human antibody response to the enzyme.

Objectives

1. Production of novel glucarpidases and glucarpidases variants by different strategies

2. Production of long acting glucarpidase(s)

3. Synthesis of Tubulysin derivatives, as a Prodrug, on a gram scale for conjugation experiments

Methods

Novel glucarpidase producers were isolated from soil using minimal plates with folate as the only carbon source. Genomic library and PCR were used to clone two novel glucarpidase genes. The two genes were overexpressed in E. coli using pET28a vector. DNA shuffling was used to produce novel glucarpidase variants. Long acting novel glucarpidase was prepared using different PEGylation techniques. Novel prodrug was prepared by novel multicomponent approach which involves the condensation of dieptide acid, β‐amino aldehyde and isocyanide derived from Cys(Trt)/OMe.

Results

We successfully produced, by DNA shuffling an ultra‐active glucarpidase that degrades MTX with a high efficiency. We also isolated three novel glucarpidase producing bacteria from Qatari soil. Two novel glucarpidases were successful cloned, overexpressed and purified. We managed to introduce a new mutation into one of the newly isolated glucarpidase gene which led to a 500% increase in the glucarpidase activity. The talk will cover many the production of long acting glucarpidase and many other issues.
Changes in Dietary Variety Scores over Six Months Among 4‐ to 8‐year‐old Zambian Children
Authors: Lewis Bess , Dyer Brian , Siamusantu Ward , Klemm Rolf , Talegawkar Sameera , Schulze Kerry , Palmer Amanda , West Keith
Published: June 2021

Dietary variety scores are often used as indicators of dietary quality, especially among women and children in resource‐poor settings. However, few studies have examined the stability of these scores over repeat measures. To study such change, we used data on dietary intakes of 4‐ to 8‐year old children living in rural Zambia (n=188) collected by 24 hour recall interviews conducted six months apart. We calculated two dietary variety scores from each 24 hour recall: a 14‐food group score and a count of individual foods. At both baseline in the late post‐harvest season and follow‐up in the early harvest season, the median food group score was 6 out of 14 [IQR (baseline and followup): 5,7) and the median food count was 8 [IQR(baseline): 6, 9.5; IQR(followup): 7, 10]. Mean absolute change from baseline to follow‐up was 1.2 for the food group score (95% CI: 1.1, 1.4) and 2.3 for the foods count (95% CI: 2.1, 2.6). Agreement between tertile classifications at baseline and follow‐up was poor (both scores: kappa<0.15). Though summary statistics of two dietary variety scores appear stable across repeat measures taken six months apart, analysis of within‐individual change reveals fluctuation in both score and tertile classification. Funded by HarvestPlus.
The Price of Oil: Behavior Change Communication and Increased Oil Ration Improve Mothers' Uses of Vegetable Oil in Preparing Food Aid Rations for Children in Malawi
Authors: Jayson Lauren , Passarelli Simone , Suri Devika , Maganga Gray , Marcus Shelley , Webb Patrick , Rogers Beatrice
Published: June 2021

Objective

As part of USAID's Food Aid Quality Review, a study evaluated the effectiveness of providing an additional oil ration in tandem with behavior change communication (BCC) to an existing supplementation program to increase the oil content and reduce sharing of porridge prepared for children under 5 with moderate acute malnutrition in Malawi.

Methods

Mothers/caretakers (BMCs) of beneficiary children in the program were randomly selected at 12 intervention and 4 control food distribution points (FDPs) for participation in interviews and to provide samples of their porridge for analysis. The intervention group received a monthly ration of 2.6 L fortified vegetable oil, 8 kg Corn Soy Blend (CSB) and BCC. Controls received a standard ration of 1 L oil, 8 kg CSB and no messaging. The proportion of BMCs preparing porridge at the target ratio (TR) of 30g oil to 100g CSB, the mean % of oil in porridge, and rates of HH sharing of porridge were compared between study groups using Stata v13.1.

Results

There were 235 BMCs in the intervention and 160 in the control groups. Roughly 6% of BMCs in the intervention group met the TR of oil to CSB, while 4% in the control group met this ratio (NS). The TR is equivalent to 23% oil; mean % oil in the prepared porridge was 21% in the intervention group and 12% in the control group (p<.001). Sharing of the porridge with other HH members was reported in 46% of intervention and 74% of control HHs (p<.001).

Conclusion

While few BMCs met the TR of oil to CSB, those receiving an additional oil ration and BCC prepared porridge with almost twice as much oil as those in the control groups, and reported less sharing within the HH than those not receiving the intervention.
Gender Differences in Body Image Perception, Weight Management Practices, and Food Choices of High School Students in the Bangkok Metropolitan Region
Authors: Chongwatpol Pitipa , Gates Gail
Published: June 2021

The objective of this study was to assess body image perceptions and weight management practices among Thai adolescents. A questionnaire was distributed to grade 10‐12 students from 9 single and mixed gender schools located in Bangkok Metropolitan Region. 2082 questionnaires with height, weight, body image perception, food choices, physical activity level, and weight management practices were included in the analysis. Approximately 55% of students had a normal BMI, while 31.2% were underweight. Only 18.1% of females and 20.6% of males did not indicate thinness dissatisfaction using Stunkard's figure rating scale (FRS). More than 66% of females selected a thinner ideal figure than their current figure, while 15.9% wanted a bigger figure. In contrast, among males, 44.3% wanted a thinner figure, but 35.1% wanted a bigger figure. However, univariate analysis observed school type but not gender differences in the degree of body image dissatisfaction; students in single gender schools had more body dissatisfaction. Females reported using more weight management practices but less physical activity than males. Males reported healthier food choices than females, and participants in single gender schools had healthier food choices compared to mixed gender schools. In conclusion, females and males had similar levels of body dissatisfaction but a majority of females want to be thinner, while males want to be either smaller or bigger. These results support the contention that it may not be appropriate to calculate an average dissatisfaction score from FRS for males because they report a preference for both smaller and bigger figures; therefore, the average dissatisfaction score will result in underreporting of body dissatisfaction.
Perspectives of Food Availability, Healthfulness and Modernity Among Adolescents in India
Authors: Shaikh Nida , Maxfield Amanda , Ramakrishnan Usha , Patil Shailaja , Cunningham Solveig
Published: June 2021

Background

Globalization of food markets has increased availability of foods & beverages (FaB) in developing regions. Adolescents' perception of FaB availability, healthfulness & modernity however is unknown.

Aim

Understand adolescents' perceptions of FaB in a globalizing Indian city.

Methods

School‐going adolescents (n=26), randomly selected, completed freelists in Bijapur, India. Participants were asked to define 'healthy, unhealthy, traditional, & modern' & freelisted FaB that sold in different food stores, new to their city, traditional & healthy, modern & healthy, traditional & unhealthy, and modern & unhealthy. Frequency & saliency for freelists were calculated.

Results

Mean age of adolescents was 15 ± 1.6 y, with 56% male & 65% attending public schools. Amongst ten most salient FaB new to city, 6 were viewed as modern & unhealthy, 3 as traditional & unhealthy, 2 as modern & healthy, and 4 as available in supermarkets & kiraana (small) stores, respectively. Six of ten most salient FaB are sold in supermarkets & kiraana stores. Vegetables & rice were among ten most salient FaB sold in supermarkets, also viewed as traditional & healthy.

Conclusion

Adolescents view healthfulness & modernity of FaB as distinct domains. Most new to city FaB believed to be modern & unhealthy are also sold at different food stores. Adolescents' perceptions of FaB need to be considered in development of interventions to improve adolescent health.
At‐Scale Delivery of Essential Nutrition Interventions (ENIs) in India is Limited by Variable Gaps in Policy, Design, Implementation and Demand
Authors: Avula Rasmi , Kim Sunny , Kohli Neha , Chakrabarti Suman , Tyagi Parul , Singh Kavita , van den Bold Mara , Kadiyala Suneetha , Menon Purnima
Published: June 2021

Evidence suggests that increasing coverage of 14 ENIs from pre‐pregnancy to early childhood can reduce maternal and child undernutrition. In India, less than 55% of women and children receive any of the ENIs. We assessed readiness to achieve at‐scale delivery of ENIs through an analysis of policies, programs, intervention design, implementation, monitoring and service use. Policy guidance and program design were examined through content analysis. We studied provision of and demand for ENIs in two states ‐ Odisha & Madhya Pradesh, using primary data collection [a survey of frontline workers (FLW) (N=1140) and households with children aged 0‐2 y (N=2300) in 3 districts per state; semi‐structured interviews with officials and health workers at multiple levels]. Evidence‐based policies and operational guidelines exist for most ENIs but not monitoring indicators; gaps exist for community‐based management of acute malnutrition (CMAM) and nutritional care during illness. Limited implementation experience exists for many ENIs, particularly for counseling of complementary feeding, pediatric anemia control and CMAM. FLW knowledge and role clarity are also weak for these ENIs. Less than 1/3 of mothers got infant feeding advice during FLW home visits; receipt of information on pediatric anemia and awareness about use of iron supplements were also low, reflecting demand challenges. Despite policies and program guidelines, scale up of ENIs in India is limited by gaps in operational guidance, monitoring indicators, unclear FLW roles and low service demand. Funding: Bill & Melinda Gates Foundation, through POSHAN, led by IFPRI
Psychosocial Determinants of Exclusive Breast‐Feeding in Rural Kenya
Authors: Gewa Constance , Chepkemboi Joan
Published: June 2021

We conducted a cross‐sectional survey to assess the association between breastfeeding‐related knowledge, perception and social norms and premature cessation of exclusive breastfeeding (EBF) in rural Kenya. Four‐hundred mothers with children, 0 ‐24 months participated in the survey. Mothers were asked to recall breastfeeding practices since birth. A battery of close and open‐ended questions were used to assess EBF‐related knowledge and social norms, and mothers' perception on the impact of EBF on children's health and growth, impact of EBF on mother's health and physical appearance and impact of EBF on family dynamics. Participants' knowledge and perception scores were calculated as a percentage of correctly‐answered questions. Mean knowledge and perception scores were above 75%. Over 14% and 23% of mothers indicated that EBF was not acceptable to their male partners and in‐laws, respectively. Higher knowledge levels and more positive perception on the impact of EBF on children's health, family dynamics and mother's health were each significantly associated with lower risks of premature cessation of EBF. Mothers who reported that EBF was not acceptable to either their male partners or in‐laws were significantly associated with higher risks of premature cessation of EBF. This study identifies specific determinants that need to be addressed in breast‐feeding promotion activities in rural Kenya. Psychosocial factors play an important role in determining individuals' decisions to engage in particular health‐related behaviors.
Effects Of Two Ready to Use Supplementary Foods (RUSF) Containing Different Levels Of Dairy Protein On Mother's Nutritional Status In Rural Guinea‐Bissau
Authors: Schlossman Nina , Batra Payal , Balan Ela , Pruzensky William , Saltzman Edward , Roberts Susan
Published: June 2021

Food insecurity and micronutrient deficiencies in Guinea‐Bissau are widespread. This Cluster Randomized Control Trial (RCT), funded by the United States Department of Agriculture, was the first such study in Guinea‐Bissau and tested the effectiveness of 2 RUSF in rural village mothers over 12 weeks.

497 mothers were randomly assigned to one of two intervention arms (92g sachets containing 500 kcal/sachet and either 15% or 33% of protein from a dairy source, provided 5 days per week at a supplement distribution center in the village) or to a control group that received no supplements. Weight, height, mid upper arm circumference (MUAC), hemoglobin, and retinol‐binding protein (RBP) were measured at baseline and endline.

Randomization to either of the intervention group resulted in improvements in maternal weight, height, body‐mass‐index (BMI), and MUAC. The intervention was correlated with reduction in self‐reported sick days. Mothers in the intervention groups experienced a smaller decrease in hemoglobin compared to control mothers (‐0.38±1.45 vs. ‐0.87±1.21, p>0.05). Study mothers are especially vulnerable to iron deficiency due to seasonal food shortages and higher iron needs among reproductive age women. The supplement appeared to have a protective effect on iron status.
Overall, the supplementation may be helpful in improving anthropometric status, preventing iron status decline, and reducing number of sick days in non‐pregnant mothers.
Resilience factors for food and nutrition security in Guatemalan communities
Authors: deFulladolsa Patricia , Rivas Adolfo , Sibrian Ricardo , Lee EunKyung , Bermudez Odilia
Published: June 2021

Objective

The interaction of poverty and climate‐related crisis, along with socio‐ecological determinants of poor nutrition, could become major factors negatively affecting food and nutrition security (FNS). Therefore, it is important to identify resilience factors to enrich FNS. This study aimed at the identification of resilience factors (RF) as contributors to food and nutrition security in 3 communities from the municipality of Santa Catarina Mita, Department of Jutiapa, Guatemala.

Methods: We assessed food and nutrition security with direct interviews, with a food security (FS) scale and with anthropometry in 184 families from the communities of La Barranca, El Quebracho y La Javia, from the southern department of Jutiapa.

Results: A total of 590 subjects, mean age of 31y with 0‐99y as range, were recruited from the selected households (hholds). FS was present in 24% of hholds. RF associated with less vs. more malnutrition in preschool age children included hholds with cement floor, using a sewer system, availability of rice, eggs and leafy leaves and head of hhold not working in agriculture. For mothers, RF for less vs. more overweight and obesity included education, not buying sweet drinks, drinking water from the public system and head of hholds growing corn and beans as cash crops.

Conclusions

Only one‐quarter of families evaluated were food secure. Contrasting them against those food insecure, we identified resilience factors to avoid malnutrition in preschoolers and to control overweight/obesity among mothers. The identification of RF for sustainable FNS and for promoting health could contribute to solutions for the socio‐economic development of rural communities.
Mothers' Child‐feeding Practices Are Associated with Children's Sugar‐sweetened Beverage Intake
Authors: Park Sohyun , Li Ruowei , Birch Leann
Published: June 2021

We examined the associations between mothers' child‐feeding practices and children's sugar‐sweetened beverage (SSB) intake among 6‐year‐olds using data from the Year 6 Follow‐Up of Infant Feeding Practices Study II (n=1350). The outcome variable was child's SSB intake. The exposure variables were 4 child‐feeding practices of mothers (set limits on sweets or junk foods; regulating child's favorite food intake to prevent overconsumption; pressuring their child eat enough; and pressuring their child to “clean the plate”). We used multivariate logistic regression and controlled for child and maternal characteristics. Analyses were stratified on child weight status. Consuming SSB 蠅1 time/day at age 6 years was observed among 17.1% of underweight/normal weight children and 23.2% of overweight/obese children. The adjusted odds for consuming SSB 蠅1 time/day (vs. no SSB intake) were significantly lower in children with mothers who set limits on sweets/junk foods (odds ratio (OR)=0.29 for underweight/normal weight children; OR=0.16 for overweight/obese children). SSB intake was higher among underweight/normal weight children whose mothers tried to keep them from eating too much of favorite foods (OR=2.03). Mothers' tendency to pressure their children to consume more food or “clean the plate” was not associated with child's SSB intake. SSBs were commonly consumed by young children and mothers' reported attempts to restrict children's intake of foods was associated with child's SSB intake. Future studies can investigate the impact of alternative child feeding practices on reducing children's SSB intake.
Plasma 3‐epi‐25‐hydroxyvitamin D3 in Very Low Birth Weight Preterm Infants in Canada During the First Five Weeks After Birth
Authors: Djekic‐Ivankovic Marija , Jones Glenville , Kaufmann Martin , Weiler Hope
Published: June 2021

Vitamin D status during infancy is important for skeletal growth and development. Response to vitamin D3 supplementation is complicated by the presence of 3‐epi‐25‐hydroxyvitamin D3 (3‐epi‐25‐OH‐D3). We undertook a small prospective study to simultaneously examine changes in 25‐OH‐D3, 3‐epi‐25‐OH‐D3 and 24,25‐dihydroxyvitamin D3 (24,25‐(OH)2D3) in preterm infants over the first five weeks after birth. Nineteen infants from Manitoba, born before 32 weeks of gestation and <1500 g, were included in the study. Plasma samples were obtained at the end of the first (wk1) and fifth (wk5) week after birth.Vitamin D metabolites were analyzed in triplicate using LC‐MS/MS. Differences over time were tested using ANOVA and Bonferroni post hoc test, p<0.05. Mean gestational age at birth was 29.0 ± 1.5 weeks and birth weight 1144.5 ± 244.4 g. Total vitamin D intake increased over time (Table). Increases were observed in vitamin D status based on plasma 25‐OH‐D3 concentration over the first 5 weeks (wk1: 18.1 ± 7.1 vs wk5: 25.3 ± 10.6 ng/ml, p=0.02) as well as in 3‐epi‐25‐OH‐D3 (wk1: 5.1 ± 4.2 vs wk5: 20.0 ± 15.9 ng/ml, p<0.0001). Over the same time‐period 24,25‐(OH)2D3 (wk1: 0.9 ± 0.5 vs wk5: 0.7 ± 0.5 ng/ml, p=0.13) was not significantly changed.This work showed that 3‐epi‐25‐OH‐D3 increased over the first 5 weeks in parallel with increases of 25‐OH‐D3 while 24,25‐(OH)2D3 was relatively stable suggesting the CYP24A1 activity does not readily mature during 5 weeks postnatally in prematurely born infants.

Vitamin D intake from all sources (IU/day)
Mean
SD
Min
Max

Week 1
202.8a
115.9
4.5
400.7

Week 2
189.3a
118.0
54.9
442.8

Week 3
250.8a
155.7
65.0
551.9

Week 4
366.2b
163.6
42.5
626.3

Week 5
489.3c
186.9
93.1
736.7

Week 6
515.9c
192.5
177.7
779.6

Means with different superscript letters are different, p<0.05