Novel methods for identification and quantification of iron fortificants in cereal flours

Iron deficiency is the most common nutrient deficiency globally. Fortification of cereal grains is a main strategy used to ameliorate global iron deficiency due to its safety and efficacy. For fortification to be effective, fortification programs must use appropriate iron compounds at appropriate levels. However, existing laboratory methods to identify and quantify fortificants are time consuming and costly. Our objective was to develop a quick and simple method to identify and quantify iron compounds commonly used for flour fortification. Unfortified whole wheat, refined wheat, and yellow corn flours were fortified with 20–60 mg Fe/kg flour using ferric pyrophosphate (FePP), ferrous sulfate (FeSO4), ferrous citrate (FeCit), ferrous fumarate (FeFum), sodium ferric EDTA (NaFeEDTA), and electrolytic iron (EFe). Using potassium thiocyanate (KSCN) with HCl with and without hydrogen peroxide (H2O2), we identified EFe, ferric, and ferrous fortificants. NaFeEDTA, FePP, FeSO4, FeCit, and FeFum were identified based on their solubility in water using ferrozine with and without ascorbic acid (ASC). An alternative method for identification that uses only KSCN as a chromogen was also developed but was inferior to the ferrozine method. Four blinded samples were prepared with randomly selected fortificants (EFe, NaFeEDTA, FePP, FeFum) and all were correctly identified by four personnel. For quantification, those four samples plus an additional FeSO4 sample were tested blindly. The average of each person\u27s reported iron levels for each sample were within 10 mg Fe/kg of actual iron levels 85% of the time. Estimated iron levels from the visual method were not significantly different than iron levels from two standard quantitative methods (p \u3e 0.05) for all the fortificants tested suggesting reliability of simple visual testing. These quick, inexpensive, and reliable methods will be useful for agencies to identify the type and amount of iron added to flour to monitor the quality of iron fortification strategies

Effects of exogenous enzymes and direct-fed microbial on broiler performance and nutrient digestibility when fed variable inclusions of soy products

The objective was to examine the effects of feed additive combinations, including xylanase, amylase, and protease enzyme (XAP), with β-mannanase (MAN) or direct-fed microbial (DFM) supplementation on broiler performance and nitrogen corrected apparent metabolizable energy (AMEn) when fed diets containing various soy products concentrations. Two experiments were conducted from 10 to 21 d of age by feeding three soy product inclusions of low (20%), intermediate (28%), and high (35%). Feed additive combinations were supplemented across the three concentrations of dietary soy in both experiments. In experiment 1 there was no enzyme supplemented, XAP (100 g/MT) alone, and XAP with MAN (100 g/MT or 50 g/MT). In experiment 2 there was no supplementation, XAP (100 g/MT) alone, and XAP with DFM (100 g/MT or 50 g/MT). Experiment 1 resulted in reduced feed intake (P ≤ 0.01) and increased body weight gain (BWG) (P ≤ 0.10) with intermediate and high soy compared to low soy inclusion. Supplementation with all enzyme combinations decreased feed intake (P ≤ 0.01) compared to non-supplemented diets and improved feed efficiency (FE; P = 0.02) and feed conversion ratio (FCR; P = 0.01). There was an interaction between soy inclusion and enzyme supplementation as XAP+½MAN improved AMEn (P = 0.05) in high soy treatments. In experiment 2, there was an interaction as the intermediate soy diet with XAP and DFM decreased BWG (P = 0.02) compared to all other soy combinations, no treatment differed from control diets without enzyme. Feed intake was not different, and high soy improved FE and FCR compared to low and intermediate soy (P ≤ 0.05). Enzyme supplementation with XAP alone improved FE and FCR over the combination of XAP and DFM (P ≤ 0.05). Low soy decreased AMEn (P ≤ 0.01) compared to intermediate and high soy. Overall, high soy diets out-performed low soy treatments possibly due to increased fiber in low soy diets from DDGS and canola meal ingredients. In experiment 1, FE and FCR were improved with all combinations of enzyme and AMEn of high soy diets was increased with enzyme supplementation demonstrating a positive effect of enzyme addition. In experiment 2, XAP and DFM did not improve broiler performance, possibly due to a well-established microbial population in the gastrointestinal tract by the time feed additive supplementation occurred

Essays on New Product Development in Emerging Markets

Global firms are increasingly moving new product development (NPD) to large emerging markets, such as India and China. In my dissertation, I study two potential NPD strategies that a global firm can pursue when entering an emerging market- (1) Shifting NPD and (2) Partnering NPD. Using a uniquely compiled panel dataset, I estimate the effect of such NPD strategies on shareholder value. In the first essay, I examine the determinants of short-term abnormal returns to a global firm’s NPD shift to an emerging market using internal resources. Investment amount (relative local employee size) is not significantly related to short-term abnormal returns. However, the effect of investment amount and relative local employee size are moderated by employee quality emphasis, costs savings emphasis, development scope and prior profitability. Employee quality emphasis has a positive moderating effect on both investment amount-- and relative local employee size-- short-term abnormal return relationships. Cost savings emphasis has a positive moderating effect on the investment amount--short-term abnormal returns relationship, but no effect on relative local employee size. Development scope (prior profitability) has a positive (negative) moderating effect on the investment amount-abnormal returns relationship. In the second essay, I investigate the determinants of the effect of NPD partnering in an emerging market on short-term abnormal returns. NPD partnering consists of a global firm engaging in NPD with a local firm through an alliance, a joint venture, or an acquisition. The findings provide actionable insights. I find that mentioning cost savings as a reason for partnering leads to negative abnormal returns. In contrast, highlighting the quality of the partner’s local employees leads to positive abnormal returns. Interestingly, the global firm’s past profitability moderates these main effects in the opposite direction. Furthermore, financial leverage has a negative effect on the short-term abnormal returns to an NPD partnership announcement. That is, the greater the global firm’s debt is relative to equity, the lower the abnormal returns are to the NPD partnership. However, a cost savings emphasis alleviates this negative effect

Evolution of an ancient protein function involved in organized multicellularity in animals.

To form and maintain organized tissues, multicellular organisms orient their mitotic spindles relative to neighboring cells. A molecular complex scaffolded by the GK protein-interaction domain (GKPID) mediates spindle orientation in diverse animal taxa by linking microtubule motor proteins to a marker protein on the cell cortex localized by external cues. Here we illuminate how this complex evolved and commandeered control of spindle orientation from a more ancient mechanism. The complex was assembled through a series of molecular exploitation events, one of which - the evolution of GKPID's capacity to bind the cortical marker protein - can be recapitulated by reintroducing a single historical substitution into the reconstructed ancestral GKPID. This change revealed and repurposed an ancient molecular surface that previously had a radically different function. We show how the physical simplicity of this binding interface enabled the evolution of a new protein function now essential to the biological complexity of many animals

Occupational Therapy Practice Patterns In Two Rural States: Does The College Experience Influence Rural Employment Choice?

An online survey was conducted of 225 occupational therapy (OT) practitioners living in the rural states of North Dakota and Wyoming to explore practice patterns and the influence of the college experience on employment choice. Findings showed that rural practitioners had greater variability of hours spent working (5.5 more hours per week, p=.028), and one more work location on average (p=.006). Therapists in urban settings spent 15% more time in fieldwork education than their rural counterparts (p=.021). Rural practice choice was influenced by participation in Level I and Level II fieldwork (p=.002) but not by loan debt. Study implications for academic programs include focusing on multiple areas of practice in the curriculum design, and exposure of students to rural practitioners and rural practice examples/experiences. Recommendations were made for rural fieldwork educator training and employer support of rural fieldwork education. Further study of the experience of working within a rural practice context as a student and OT practitioner are recommended, including variables impacting rural practitioner work with fieldwork students, and student interest in rural fieldwork placement

The epidemiology and outcomes of central nervous system infections in Far North Queensland, tropical Australia; 2000-2019

Background: The epidemiology of central nervous system (CNS) infections in tropical Australia is incompletely defined. Methods: A retrospective study of all individuals in Far North Queensland, tropical Australia, who were diagnosed with a CNS infection between January 1, 2000, and December 31, 2019. The microbiological aetiology of the infection was correlated with patients' demographic characteristics and their clinical course. Results: There were 725 cases of CNS infection during the study period, meningitis (77.4%) was the most common, followed by brain abscess (11.6%), encephalitis (9.9%) and spinal infection (1.1%). Infants (24.3%, p<0.0001) and Aboriginal and Torres Strait Islander Australians (175/666 local residents, 26.3%, p<0.0001) were over-represented in the cohort. A pathogen was identified in 513 cases (70.8%); this was viral in 299 (41.2%), bacterial in 175 (24.1%) and fungal in 35 (4.8%). Cryptococcal meningitis (24 cases) was diagnosed as frequently as pneumococcal meningitis (24 cases). There were only 2 CNS infections with a S. pneumoniae serotype in the 13-valent pneumococcal vaccine after its addition to the National Immunisation schedule in 2011. Tropical pathogens-including Cryptococcus species (9/84, 11%), Mycobacterium tuberculosis (7/84, 8%) and Burkholderia pseudomallei (5/ 84, 6%)-were among the most common causes of brain abscess. However, arboviral CNS infections were rare, with only one locally acquired case-a dengue infection in 2009-diagnosed in the entire study period. Intensive Care Unit admission was necessary in 14.3%; the overall case fatality rate was 4.4%. Conclusion: Tropical pathogens cause CNS infections as commonly as traditional bacterial pathogens in this region of tropical Australia. However, despite being highlighted in the national consensus guidelines, arboviruses were identified very rarely. Prompt access to sophisticated diagnostic and supportive care in Australia's well-resourced public health system is likely to have contributed to the cohort's low case-fatality rate

Citizen-led sampling to monitor phosphate levels in freshwater environments using a simple paper microfluidic device

Contamination of waterways is of increasing concern, with recent studies demonstrating elevated levels of antibiotics, antidepressants, household, agricultural and industrial chemicals in freshwater systems. Thus, there is a growing demand for methods to rapidly and conveniently monitor contaminants in waterways. Here we demonstrate how a combination of paper microfluidic devices and handheld mobile technology can be used by citizen scientists to carry out a sustained water monitoring campaign. We have developed a paper-based analytical device and a 3 minute sampling workflow that requires no more than a container, a test device and a smartphone app. The contaminant measured in these pilots are phosphates, detectable down to 3 mg L-1. Together these allow volunteers to successfully carry out cost-effective, high frequency, phosphate monitoring over an extended geographies and periods

Phage Display Approaches for the Isolation of Monoclonal Antibodies Against Dengue Virus Envelope Domain III from Human and Mouse Derived Libraries

Domain III of the dengue virus envelope protein (EDIII, aa295-395) has an immunoglobulin fold and is the proposed receptor-binding domain of the virus. Previous studies have shown that monoclonal antibodies against EDIII can be neutralizing and have therapeutic potential. Here, cloned Fab-phage libraries of human and mouse origin were screened for DENV specific antibodies. Firstly, bacterially expressed EDIII or whole virus particles were used as bait in biopanning against a large naïve human Fab-phage library (>10 billion independent clones). Multiple panning strategies were employed, and in excess of 1000 clones were screened, but all of the antibodies identified bound the envelope in regions outside EDIII suggesting EDIII antibodies are virtually absent from the naïve human repertoire. Next, a chimeric Fab-phage library was constructed from a panel of EDIII specific mouse hybridomas by pooling the VH and VL chain sequences from the hybridomas and cloning these into the pComb3X phagemid vector with human CH and CL encoding sequences. Biopanning against EDIII identified a unique antibody (C9) that cross-reacts with EDIII from DENV1-3 and, in the IgG format, binds and neutralizes DENV2 in cell-based assays. Sequence analysis and saturation mutagenesis of complementary determining regions (CDR) in the C9 light chain suggest an antigen recognition model in which the LCDR3 is a key determinant of EDIII specificity, while modifications in LCDR1 and LCDR2 affect DENV serotype cross-reactivity. Overall, this study supports the current prevailing opinion that neutralizing anti-EDIII monoclonal antibodies can be readily generated in murine systems, but in humans the anti-DENV immune response is directed away from domain III

Supporting surveillance capacity for antimicrobial resistance: Laboratory capacity strengthening for drug resistant infections in low and middle income countries.

Development of antimicrobial resistance (AMR) threatens our ability to treat common and life threatening infections. Identifying the emergence of AMR requires strengthening of surveillance for AMR, particularly in low and middle-income countries (LMICs) where the burden of infection is highest and health systems are least able to respond. This work aimed, through a combination of desk-based investigation, discussion with colleagues worldwide, and visits to three contrasting countries (Ethiopia, Malawi and Vietnam), to map and compare existing models and surveillance systems for AMR, to examine what worked and what did not work. Current capacity for AMR surveillance varies in LMICs, but and systems in development are focussed on laboratory surveillance. This approach limits understanding of AMR and the extent to which laboratory results can inform local, national and international public health policy. An integrated model, combining clinical, laboratory and demographic surveillance in sentinel sites is more informative and costs for clinical and demographic surveillance are proportionally much lower. The speed and extent to which AMR surveillance can be strengthened depends on the functioning of the health system, and the resources available. Where there is existing laboratory capacity, it may be possible to develop 5-20 sentinel sites with a long term view of establishing comprehensive surveillance; but where health systems are weaker and laboratory infrastructure less developed, available expertise and resources may limit this to 1-2 sentinel sites. Prioritising core functions, such as automated blood cultures, reduces investment at each site. Expertise to support AMR surveillance in LMICs may come from a variety of international, or national, institutions. It is important that these organisations collaborate to support the health systems on which AMR surveillance is built, as well as improving technical capacity specifically relating to AMR surveillance. Strong collaborations, and leadership, drive successful AMR surveillance systems across countries and contexts

Differences in Maternal and Infant Cord Blood Vitamin D Between Racial/Ethnic Groups

Background: Vitamin D deficiency associated with lower 25-hydroxyvitamin D (25(OH)D) concentration is common among individuals with more melanin pigmentation. Low 25(OH)D levels in pregnant women may be related to increased risk of low birth weight and preterm delivery. Still, few studies have assessed how serum levels of 25(OH)D vary between maternal and infant race/ethnicity. Objective: This study aimed to investigate the relationship between 25(OH)D levels in maternal blood and infant cord blood within certain ethnic groups, prematurity status, and low birth weight. Experimental Design:An IRB-approved study enrolled 86 mother-infant pairs. Maternal blood samples and infant cord blood samples were analyzed for 25(OH)D serum levels. Descriptive statistics and Kruskal-Willis tests comparisons were conducted with the use of IBM SPSS Statistics 28 software to assess the relationship between maternal and cord blood 25(OH)D levels in other race/ethnicity groups, birthweight, and preterm birth. Prematurity was categorized into two groups: premature (weeks) and term (≥37 weeks). Birth weight was categorized into two groups: low birth weight (\u3c 2500 g weeks) and not low birth weight (≥2500 g weeks). A p-value of Results:Median levels of 25(OH)D serum were lower in infant’s cord blood (22.52 ng/mL) than maternal blood (38.06 ng/mL). White participants had significantly higher 25(OH)D levels than African American participants in both maternal blood (40.76 ng/mL vs 27.79, p = Conclusion: Our findings suggest a possible association with lower serum 25-hydroxyvitamin D concentration in darker skin pigmentation, even in a small sample size. These results suggest that prematurity and birth weight should be replicated in larger sample sizes of different Race/Ethnic groups, limiting this finding. Further studies should focus on examining differences with larger and more diverse sample sizes. Such research should include measuring Vitamin D intake in pregnancy and clinical outcomes.https://digitalcommons.unmc.edu/surp2021/1008/thumbnail.jp