Development and validation of a quantitative choline food frequency questionnaire for use with drinking and non-drinking pregnant women in Cape Town,South Africa

Background

Although animal and human studies have demonstrated interactions between dietary choline and fetal alcohol spectrum disorders, dietary choline deficiency in pregnancy is common in the US and worldwide. We sought to develop and validate a quantitative food frequency questionnaire (QFFQ) to estimate usual daily choline intake in pregnant mothers.

Methods

A panel of nutrition experts developed a Choline-QFFQ food item list, including sources with high choline content and the most commonly consumed choline-containing foods in the target population. A data base for choline content of each item was compiled. For reliability and validity testing in a prospective longitudinal cohort, 123 heavy drinking Cape Coloured pregnant women and 83 abstaining/light-drinking controls were recruited at their first antenatal clinic visit. At 3 prenatal study visits, each gravida was interviewed about alcohol, smoking, and drug use, and administered a 24-hour recall interview and the Choline-QFFQ.

Results

Across all visits and assessments, > 78% of heavy drinkers and controls reported choline intake below the Dietary Reference Intakes adequate intake level (450 mg/day). Women reported a decrease in choline intake over time on the QFFQ. Reliability of the QFFQ across visits was good-to-acceptable for 2 of 4 group-level tests and 4 of 5 individual-level tests for both drinkers and controls. When compared with 24-hr recall data, validity of the QFFQ was good-to-acceptable for 3 of 4 individual-level tests and 3 of 5 group-level tests. For controls, validity was good-to-acceptable for all 4 individual-level tests and all 5 group-level tests.

Conclusions

To our knowledge, this is the first quantitative choline food frequency screening questionnaire to be developed and validated for use with both heavy and non-drinking pregnant women and the first to be used in the Cape Coloured community in South Africa. Given the high prevalence of inadequate choline intake and the growing evidence that maternal choline supplementation can mitigate some of the adverse effects of prenatal alcohol exposure, this tool may be useful for both research and future clinical outreach programs.

     

Stereological estimation of the total volume of MR visible brain lesions in patients with multiple sclerosis

Point counting represents a convenient and efficient technique for estimating the area of transects through multiple sclerosis (MS) lesions on magnetic resonance (MR) images obtained for sections through the brain. When sectioning has been performed according to the Cavalieri method, unbiased estimates of the total volume of MR-visible MS plaques can be obtained with a precision of 3–5% in 5–10 min.     

NMR analysis of butyl acrylate/methylmethacrylate/α‐methyl styrene terpolymers

Composition analysis for butyl acrylate (BA)/ methyl methacrylate (MMA)/α‐methyl styrene terpolymers was carried out by NMR spectroscopy methods. ¹H‐NMR was used primarily for this analysis, but because the method did not provide independent measurements for the BA and MMA fractions, the terpolymer composition analysis was open to higher than normal levels of uncertainty. Supplementary analyses were made with quantitative ¹³C‐NMR methods to confirm the results from ¹H‐NMR (quantitative ¹³C‐NMR was used to provide corroboration of selected composition analyses). To confirm spectral assignments in the ¹H‐ and ¹³C‐NMR spectra, heteronuclear multiple quantum coherence and J Modulated Spin Echo (JMOD) pulse sequences were used. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 103: 2093–2098, 2007     

Quantitative probing the interfacial structure of TPO/CPO blends by transmission electron microscopy via EDX

High-resolution scanning transmission electron microscope (HR STEM) measurements were performed on a thermoplastic polyolefin (TPO) substrate coated with chlorinated polyolefin (CPO). This CPO was a maleated chlorinated polypropylene containing 21.8 wt% Cl. The TPO investigated was a blend of high-modulus isotactic polypropylene (iPP) with a crystalline ethylene-butene copolymer (EB9) containing 9 wt% butene. For these injection-molded samples, examined ca. 10 mm from the mold gate, a stratified morphology found at TPO surface consisting of thin fibers of EB9 trapped in a transcrystalline iPP matrix, with crystalline lamellae propagating from the matrix across the EB9 domains. This structure was unperturbed when the plaques were coated (from tetrahydrofuran solution) with a 5 μm layer of CPO, but underwent changes of increasing severity when subjected either to a dry bake at 120 °C or annealing at 120 °C in the presence of xylene vapor. The interfacial structure between the CPO and the TPO was probed by TEM with energy dispersive X-ray imaging (EDX). The elemental chlorine across the interface gave good fits to a tan h function, and the interface thickness increased from 23 ± 2 nm to 28 ± 1 nm upon annealing at 120 °C for 30 min. After annealing in the presence of xylene vapor, this value increased to 50 ± 4 nm. As reference points, we determined an interface thickness of 29 ± 3 nm for the CPO-EB interface and 15 ± 2 nm for the interface between CPO and iPP.     

Effect of Temperatures up to 90°C on the Early Hydration of Portland–Blastfurnace Slag Cements

Isothermal conduction calorimetry has been used to monitor the early hydration of Portland–blastfurnace slag (BFS)-blended cements. Portland:BFS composite cements with ordinary Portland cement replacements from 0 to 90 wt% were studied at curing temperatures from 12° to 90°C. Peak II, principally associated with alite (Ca₃SiO₅) hydration, was accelerated with increasing temperature for all blends. Peak S, associated with BFS hydration, was particularly noticeable at 40° and 60°C. At higher curing temperatures, peak S merged with peak II, indicating thermal activation of BFS. Novel plots of total heat output against percentage replacement show that BFS contributes to the heat of hydration, even at temperatures below its thermal activation.     

Effect of ionic strength on the reactivity ratios of acrylamide/acrylic acid (sodium acrylate) copolymerization

The ionic strength (IS) of polyelectrolyte solutions plays an important role in influencing reaction kinetics. The largely unstudied effect of IS on monomer reactivity ratios and copolymerization rates of acrylamide (AAm) and acrylic acid (AAc), in the form of sodium acrylate (NaAc), is investigated. Salt addition affects the nature of overall charges of the polyelectrolyte solution and diminishes the electrostatic repulsions between reacting chains. Therefore, changing the IS of the solution by incorporating salts affect not only the point estimates of the monomer reactivity ratios but also the overall behavior of the copolymerization (with a transition to azeotropic behavior). Experimental results on copolymerization rates confirm the observed trends in reactivity ratio behavior. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40949.     

Colloidal Stability & Conformational Changes in β-Lactoglobulin: Unfolding to Self-Assembly

A detailed understanding of the mechanism of unfolding, aggregation, and associated rheological changes is developed in this study for β-Lactoglobulin at different pH values through concomitant measurements utilizing dynamic light scattering (DLS), optical microrheology, Raman spectroscopy, and differential scanning calorimetry (DSC). The diffusion interaction parameter k_D emerges as an accurate predictor of colloidal stability for this protein consistent with observed aggregation trends and rheology. Drastic aggregation and gelation were observed at pH 5.5. Under this condition, the protein’s secondary and tertiary structures changed simultaneously. At higher pH (7.0 and 8.5), oligomerizaton with no gel formation occurred. For these solutions, tertiary structure and secondary structure transitions were sequential. The low frequency Raman data, which is a good indicator of hydrogen bonding and structuring in water, has been shown to exhibit a strong correlation with the rheological evolution with temperature. This study has, for the first time, demonstrated that this low frequency Raman data, in conjunction with the DSC endotherm, can be been utilized to deconvolve protein unfolding and aggregation/gelation. These findings can have important implications for the development of protein-based biotherapeutics, where the formulation viscosity, aggregation, and stability strongly affects efficacy or in foods where protein structuring is critical for functional and sensory performance.     

Substrate Flexibility of the Flavin-Dependent Dihydropyrrole Oxidases PigB and HapB Involved in Antibiotic Prodigiosin Biosynthesis

In the biosynthesis of the tripyrrolic pigment prodigiosin, PigB is a predicted flavin-dependent oxidase responsible for the formation of 2-methyl-3-amylpyrrole (MAP) from a dihydropyrrole. To prove which dihydropyrrole is the true intermediate, both possibilities, 5-methyl-4-pentyl-3,4-dihydro-2H-pyrrole ( 5 a , resulting from transamination of the aldehyde of 3-acetyloctanal) and 2-methyl-3-pentyl-3,4-dihydro-2H-pyrrole ( 6 , resulting from transamination of the ketone), were synthesised. Only 5 a restored pigment production in a strain of Serratia sp. ATCC 39006 blocked earlier in MAP biosynthesis. PigB is membrane-associated and inactive when its transmembrane domain was deleted, but HapB, its homologue in Hahella chejuensis, lacks the transmembrane domain and is active in solution. Two colourimetric assays for PigB and HapB were developed, and the HapB-catalysed reaction was kinetically characterised. Ten analogues of 5 a were synthesised, varying in the C2 and C3 side chains, and tested as substrates of HapB in vitro and for restoration of pigment production in Serratia ΔpigD in vivo. All lengths of side chain tested at C3 were accepted, but only short side chains at C2 were accepted. The knowledge that 5 a is an intermediate in prodigiosin biosynthesis and the ease of synthesis of analogues of 5 a makes a range of prodigiosin analogues readily available by mutasynthesis.     

Click-Chemistry (CuAAC) Trimerization of an αvβ6 Integrin Targeting Ga-68-Peptide: Enhanced Contrast for in-Vivo PET Imaging of Human Lung Adenocarcinoma Xenografts

α_vβ₆ Integrin is an epithelial transmembrane protein that recognizes latency-associated peptide (LAP) and primarily activates transforming growth factor beta (TGF-β). It is overexpressed in carcinomas (most notably, pancreatic) and other conditions associated with α_vβ₆ integrin-dependent TGF-β dysregulation, such as fibrosis. We have designed a trimeric Ga-68-labeled TRAP conjugate of the α_vβ₆-specific cyclic pentapeptide SDM17 (cyclo[RGD-Chg-E]-CONH₂) to enhance α_vβ₆ integrin affinity as well as target-specific in-vivo uptake. Ga-68-TRAP(SDM17)₃ showed a 28-fold higher α_vβ₆ affinity than the corresponding monomer Ga-68-NOTA-SDM17 (IC₅₀ of 0.26 vs. 7.4 nM, respectively), a 13-fold higher IC₅₀-based selectivity over the related integrin α_vβ₈ (factors of 662 vs. 49), and a threefold higher tumor uptake (2.1 vs. 0.66 %ID/g) in biodistribution experiments with H2009 tumor-bearing SCID mice. The remarkably high tumor/organ ratios (tumor-to-blood 11.2; -to-liver 8.7; -to-pancreas 29.7) enabled high-contrast tumor delineation in PET images. We conclude that Ga-68-TRAP(SDM17)₃ holds promise for improved clinical PET diagnostics of carcinomas and fibrosis.