Form and Function: X-Ray Scattering and Spectroscopy of Transition Metal-Based Nanoparticles

In recent decades, nanoparticles have been found to possess unique, tunable properties with an enormous variety of applications. The atomic and nanoscale structures govern these functional properties, and structural deviations from the bulk, in part, are responsible for the vast technological uses of nanoparticles. This dissertation tackles the understanding of structure in a number of metal, metal phosphide, and metal oxide nanoparticle systems. Additionally, the syntheses of monodispersed nanoparticle systems allow for correlating their structure with functional properties. Real space analysis using pair distribution functions of monometallic (Ni, Pd) nanoparticles of less than 5 nm in diameter revealed a deviation from the bulk face-centered cubic structure. Their local atomic packing disorder and lack of long-range order resemble that of bulk metallic glasses, which often consist of complex mixtures of a multitude of elements. Bulk metallic glasses have high mechanical strength and can sustain elastic deformations. The significant connection between these two seemingly disparate systems lie in the short-range ordering of their atomic packing motifs, which consist of icosahedral symmetry as seen in their pair distribution functions. Cobalt phosphide (Co2P) nanorods are promising as inexpensive, earth abundant catalysts for the oxygen reduction reaction in fuel cells. Additionally, their 1-D structures demonstrated greater stability as compared to conventional Pt catalysts. Their structure was investigated using high-resolution electron microscopy and a suite of X-ray scattering and absorption techniques. The dynamic structural nature of the solid-solid phase transition in vanadium dioxide (VO2) thin films was investigated using X-ray absorption fine structure spectroscopy. Substitution of transition metal dopants into lattice sites revealed the structurally-driven depression of the metal-to-insulator transition temperature. Bridging form and function, this dissertation reports the colloidal synthesis of monodispersed nanoparticles alongside structural investigations and functional testing

Formation and Degradation of Beta-casomorphins in Dairy Processing

Milk proteins including casein are sources of peptides with bioactivity. One of these peptides is beta-casomorphin (BCM) which belongs to a group of opioid peptides formed from b-casein variants. Beta-casomorphin 7 (BCM7) has been demonstrated to be enzymatically released from the A1 or B b-casein variant. Epidemiological evidence suggests the peptide BCM 7 is a risk factor for development of human diseases, including increased risk of type 1 diabetes and cardiovascular diseases but this has not been thoroughly substantiated by research studies. High performance liquid chromatography coupled to UV-Vis and mass spectrometry detection as well as enzyme–linked immunosorbent assay (ELISA) has been used to analyze BCMs in dairy products. BCMs have been detected in raw cow’s milk and human milk and a variety of commercial cheeses, but their presence has yet to be confirmed in commercial yoghurts. The finding that BCMs are present in cheese suggests they could also form in yoghurt, but be degraded during yoghurt processing. Whether BCMs do form in yoghurt and the amount of BCM forming or degrading at different processing steps needs further investigation and possibly will depend on the heat treatment and fermentation process used, but it remains an intriguing unknown

Familial hypercholesterolaemia in children and adolescents from 48 countries: a cross-sectional study

Background: Approximately 450 000 children are born with familial hypercholesterolaemia worldwide every year, yet only 2·1% of adults with familial hypercholesterolaemia were diagnosed before age 18 years via current diagnostic approaches, which are derived from observations in adults. We aimed to characterise children and adolescents with heterozygous familial hypercholesterolaemia (HeFH) and understand current approaches to the identification and management of familial hypercholesterolaemia to inform future public health strategies. Methods: For this cross-sectional study, we assessed children and adolescents younger than 18 years with a clinical or genetic diagnosis of HeFH at the time of entry into the Familial Hypercholesterolaemia Studies Collaboration (FHSC) registry between Oct 1, 2015, and Jan 31, 2021. Data in the registry were collected from 55 regional or national registries in 48 countries. Diagnoses relying on self-reported history of familial hypercholesterolaemia and suspected secondary hypercholesterolaemia were excluded from the registry; people with untreated LDL cholesterol (LDL-C) of at least 13·0 mmol/L were excluded from this study. Data were assessed overall and by WHO region, World Bank country income status, age, diagnostic criteria, and index-case status. The main outcome of this study was to assess current identification and management of children and adolescents with familial hypercholesterolaemia. Findings: Of 63 093 individuals in the FHSC registry, 11 848 (18·8%) were children or adolescents younger than 18 years with HeFH and were included in this study; 5756 (50·2%) of 11 476 included individuals were female and 5720 (49·8%) were male. Sex data were missing for 372 (3·1%) of 11 848 individuals. Median age at registry entry was 9·6 years (IQR 5·8-13·2). 10 099 (89·9%) of 11 235 included individuals had a final genetically confirmed diagnosis of familial hypercholesterolaemia and 1136 (10·1%) had a clinical diagnosis. Genetically confirmed diagnosis data or clinical diagnosis data were missing for 613 (5·2%) of 11 848 individuals. Genetic diagnosis was more common in children and adolescents from high-income countries (9427 [92·4%] of 10 202) than in children and adolescents from non-high-income countries (199 [48·0%] of 415). 3414 (31·6%) of 10 804 children or adolescents were index cases. Familial-hypercholesterolaemia-related physical signs, cardiovascular risk factors, and cardiovascular disease were uncommon, but were more common in non-high-income countries. 7557 (72·4%) of 10 428 included children or adolescents were not taking lipid-lowering medication (LLM) and had a median LDL-C of 5·00 mmol/L (IQR 4·05-6·08). Compared with genetic diagnosis, the use of unadapted clinical criteria intended for use in adults and reliant on more extreme phenotypes could result in 50-75% of children and adolescents with familial hypercholesterolaemia not being identified. Interpretation: Clinical characteristics observed in adults with familial hypercholesterolaemia are uncommon in children and adolescents with familial hypercholesterolaemia, hence detection in this age group relies on measurement of LDL-C and genetic confirmation. Where genetic testing is unavailable, increased availability and use of LDL-C measurements in the first few years of life could help reduce the current gap between prevalence and detection, enabling increased use of combination LLM to reach recommended LDL-C targets early in life

Application of ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry (Orbitrap™) for the determination of beta-casein phenotypes in cow milk

The objective of this study was to determine the β-CN phenotypes in cow milk collected from HF and cross-bred HF dairy cattle in Phu Dong, Vietnam. In total, 85 samples of raw milk were collected from 85 individual cows. Beta-casein (β-CN) phenotypes in cow milk were determined using ultra-high performance liquid chromatography coupled to high resolution mass spectrometry (UHPLC-HRMS). The results showed that three β-CN variants A1, A2 and I were detected and identified in the milk samples. Five β-CN phenotypes A1A1, A1A2, A1I, A2A2 and A2I were found with percentages of 0.035, 0.400, 0.059, 0.482 and 0.024, respectively. The higher proportion of β-CN phenotype A2A2 compared to other phenotypes was expected because of changes in dairy cow breeding in Phu Dong, Vietnam

Identification and quantification of beta-casomorphin peptides naturally yielded in raw milk by liquid chromatography-tandem mass spectrometry

Beta-casomorphin peptides including beta-casomorphin 5 (β-CM5) and beta-casomorphin 7 (β-CM7) in raw cow milk were analyzed using a liquid chromatography-tandem mass spectrometry (LC-MS/MS). Milk samples were acidified with HCl followed by centrifuged and cleaned-up using solid-phase extraction (SPE). The eluents were evaporated to dryness and subsequently reconstituted with methanol. Limits of detection (LOD) were 0.32 ng/g for β-CM5 and 0.24 ng/g for β-CM7 and limits of quantification (LOQ) were 1.06 ng/g and 0.69 ng/g for β-CM5 and β-CM7, respectively. Recoveries ranged from 89.7% to 99.4% for β-CM5 and from 96.3% to 98.9% for β-CM7. Intra-day precision was ranged between 7.3% and 22.7% for β-CM5; and 4.5% and 7.3% for β-CM7, meanwhile inter-day precision resulted 7.3%–16.5% for β-CM5 and 9.3%–12.8% for β-CM7. The concentration of β-CM5/7 was analyzed in different milk samples. β-CM7 was detected in 21 out of 30 samples and ranged from 0.76 to 8.41 ng/g milk, while β-CM5 was unequivocally identified in 3 samples, its concentration was 0.4–0.64 ng/g. This is the first report to describe the natural occurrence of β-CM5/7 in raw milk using LC-MS/MS. The present study provides evidence of β-CMs, especially β-CM7 that can be released in raw milk from different breeds