Search

Maternal smoking during pregnancy (MSDP), driven by nicotine crossing the placenta, causes lifelong decreases in offspring pulmonary function and vitamin C supplementation during pregnancy prevents some of those changes. We have also shown in animal models of prenatal nicotine exposure that vitamin C supplementation during pregnancy improves placental function. In this study we examined whether vitamin C supplementation mitigates the effects of MSDP on placental structure, function, and gene expression in pregnant human smokers. Doppler ultrasound was performed in a subset of 55 pregnant smokers participating in the "Vitamin C to Decrease the Effects of Smoking in Pregnancy on Infant Lung Function" (VCSIP) randomized clinical trial (NCT01723696) and in 33 pregnant nonsmokers. Doppler ultrasound measurements showed decreased umbilical vein Doppler velocity (Vmax) in placebo-treated smokers that was significantly improved in smokers randomized to vitamin C, restoring to levels comparable to nonsmokers. RNA-sequencing demonstrated that vitamin C supplementation to pregnant smokers was associated with changes in mRNA expression in genes highly relevant to vascular and cardiac development, suggesting a potential mechanism for vitamin C supplementation in pregnant smokers to improve some aspects of offspring health., (© 2024. The Author(s).)

Ferrous iron (Fe 2+ ) transport is an essential process that supports the growth, intracellular survival, and virulence of several drug-resistant pathogens, and the ferrous iron transport (Feo) system is the most important and widespread protein complex that mediates Fe 2+ transport in these organisms. The Feo system canonically comprises three proteins (FeoA/B/C). FeoA and FeoC are both small, accessory proteins localized to the cytoplasm, and their roles in the Fe 2+ transport process have been of great debate. FeoB is the only wholly-conserved component of the Feo system and serves as the inner membrane-embedded Fe 2+ transporter with a soluble G-protein-like N-terminal domain. In vivo studies have underscored the importance of Feo during infection, emphasizing the need to better understand Feo-mediated Fe 2+ uptake, although a paucity of research exists on intact FeoB. To surmount this problem, we designed an overproduction and purification system that can be applied generally to a suite of intact FeoBs from several organisms. Importantly, we noted that FeoB is extremely sensitive to excess salt while in the membrane of a recombinant host, and we designed a workflow to circumvent this issue. We also demonstrated effective protein extraction from the lipid bilayer through small-scale solubilization studies. We then applied this approach to the large-scale purifications of Escherichia coli and Pseudomonas aeruginosa FeoBs to high purity and homogeneity. Lastly, we show that our protocol can be generally applied to various FeoB proteins. Thus, this workflow allows for isolation of suitable quantities of FeoB for future biochemical and biophysical characterization., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Exposure to alcohol has multiple effects on nervous system function, and organisms have evolved mechanisms to optimally respond to the presence of ethanol. Sex differences in ethanol-induced behaviors have been observed in several organisms, ranging from humans to invertebrates. However, the molecular mechanisms underlying the dimorphic regulation of ethanol-induced behaviors remain incompletely understood. Here, we observed sex differences in ethanol sedation sensitivity in Drosophila Genome Reference Panel (DGRP) lines of Drosophila melanogaster compared to the absence of dimorphism in standard laboratory wildtype and control lines. However, in dose response experiments, we were able to unmask dimorphic responses for the control mutant line w 1118 by lowering the testing ethanol concentration. Notably, feminization of the small population of Corazonin (Crz) neurons in males was sufficient to induce female-like sedation sensitivity. We also tested the role of the transcription factor apontic ( apt ) based on its known expression in Crz neurons and its regulation of sedation responses. Interestingly, loss of function apt mutations increased sedation times in both males and females as compared to controls. No significant difference between male and female apt mutants was observed, suggesting a possible role of apt in the regulation of dimorphic ethanol-induced responses. Thus, our results shed light into the mechanisms regulating sex-differences in ethanol-induced behaviors at the cellular and molecular level, suggesting that the genetic sex in a small neuronal population plays an important role in modulating sex differences in behavioral responses to ethanol., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Oyeyinka, Kansal, O’Sullivan, Gualtieri, Smith and Vonhoff.)

Iron is an essential element for nearly all organisms, and under anoxic and/or reducing conditions, Fe 2+ is the dominant form of iron available to bacteria. The ferrous iron transport (Feo) system is the primary prokaryotic Fe 2+ import machinery, and two constituent proteins (FeoA and FeoB) are conserved across most bacterial species. However, how FeoA and FeoB function relative to one another remains enigmatic. In this work, we explored the distribution of feoAB operons encoding a fusion of FeoA tethered to the N-terminal, G-protein domain of FeoB via a connecting linker region. We hypothesized that this fusion poises FeoA to interact with FeoB to affect function. To test this hypothesis, we characterized the soluble NFeoAB fusion protein from Bacteroides fragilis, a commensal organism implicated in drug-resistant infections. Using X-ray crystallography, we determined the 1.50-Å resolution structure of BfFeoA, which adopts an SH3-like fold implicated in protein-protein interactions. Using a combination of structural modeling, small-angle X-ray scattering, and hydrogen-deuterium exchange mass spectrometry, we show that FeoA and NFeoB interact in a nucleotide-dependent manner, and we mapped the protein-protein interaction interface. Finally, using guanosine triphosphate (GTP) hydrolysis assays, we demonstrate that BfNFeoAB exhibits one of the slowest known rates of Feo-mediated GTP hydrolysis that is not potassium-stimulated. Importantly, truncation of FeoA from this fusion demonstrates that FeoA-NFeoB interactions function to stabilize the GTP-bound form of FeoB. Taken together, our work reveals a role for FeoA function in the fused FeoAB system and suggests a function for FeoA among prokaryotes., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Maternal smoking during pregnancy (MSDP) affects development of multiple organ systems including the placenta, lung, brain, and vasculature. In particular, children exposed to MSDP show lifelong deficits in pulmonary function and increased risk of asthma and wheeze. Our laboratory has previously shown that vitamin C supplementation during pregnancy prevents some of the adverse effects of MSDP on offspring respiratory outcomes. Epigenetic modifications, including DNA methylation (DNAm), are a likely link between in utero exposures and adverse health outcomes, and MSDP has previously been associated with DNAm changes in blood, placenta, and buccal epithelium. Analysis of placental DNAm may reveal critical targets of MSDP and vitamin C relevant to respiratory health outcomes., Results: DNAm was measured in placentas obtained from 72 smokers enrolled in the VCSIP RCT: NCT03203603 (37 supplemented with vitamin C, 35 with placebo) and 24 never-smokers for reference. Methylation at one CpG, cg20790161, reached Bonferroni significance and was hypomethylated in vitamin C supplemented smokers versus placebo. Analysis of spatially related CpGs identified 93 candidate differentially methylated regions (DMRs) between treatment groups, including loci known to be associated with lung function, oxidative stress, fetal development and growth, and angiogenesis. Overlap of nominally significant differentially methylated CpGs (DMCs) in never-smokers versus placebo with nominally significant DMCs in vitamin C versus placebo identified 9059 candidate "restored CpGs" for association with placental transcript expression and respiratory outcomes. Methylation at 274 restored candidate CpG sites was associated with expression of 259 genes (FDR < 0.05). We further identified candidate CpGs associated with infant lung function (34 CpGs) and composite wheeze (1 CpG) at 12 months of age (FDR < 0.05). Increased methylation in the DIP2C, APOH/PRKCA, and additional candidate gene regions was associated with improved lung function and decreased wheeze in offspring of vitamin C-treated smokers., Conclusions: Vitamin C supplementation to pregnant smokers ameliorates changes associated with maternal smoking in placental DNA methylation and gene expression in pathways potentially linked to improved placental function and offspring respiratory health. Further work is necessary to validate candidate loci and elucidate the causal pathway between placental methylation changes and outcomes of offspring exposed to MSDP. Clinical trial registration ClinicalTrials.gov, NCT01723696. Registered November 6, 2012. https://clinicaltrials.gov/ct2/show/record/NCT01723696 ., (© 2021. The Author(s).)

Educational systems across the globe were disrupted by the COVID-19 pandemic, and faculty, staff, and students had to rapidly transition to e-learning platforms. These groups had little preparation to cope with the challenges of this newly adopted system. However, as we begin to emerge from the COVID-19 era, efforts are being made to assess the impact of this transition and develop a framework of best practices to help educators prepare for possible future disruptions. This commentary aims to discuss some of the challenges associated with the rapid transition to the new academic environment, including the modes of instruction employed, technical obstacles encountered, student responses to change and efforts made to evaluate didactic and practical aspects of the curriculum in the contexts of premedical and medical education, at the newly established College of Medicine at Khalifa University of Science and Technology in the United Arab Emirates., Competing Interests: Declaration of conflicting interests: The author declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2021.)

The authors recount the challenges they overcame to deliver lecture content and assessments while engaging students at their newly established medical school. Faculty must multitask in new and added ways to achieve the same goal in e-learning platforms. Online course delivery introduces additional barriers to engaging students, atypical of face-to-face sessions. We received valuable feedback, adjusted our delivery, and allowed our students to access lecture recordings at their convenience. Our sessions with students were more than just a lecture but a way to help people through a unprecedented time. Remote learning platforms also provided faculty with opportunities to develop new pedagogical skills and alternative assessments., Competing Interests: Declaration of conflicting interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2021.)

National nutrition surveys have shown that over half of all adults in Ireland, the United Kingdom (UK), and the United States of America (USA) have low vitamin K intakes. Thus, dietary strategies to improve vitamin K intakes are needed, and vitamin K biofortification of food may be one food-based approach. The primary aim of our study was to establish whether increasing the vitamin K 3 content of hen feed can increase the vitamin K content of eggs, and the secondary aims were to examine the effects on hen performance parameters, as well as egg and eggshell quality parameters. A 12 week hen feeding trial was conducted in which Hyline chickens were randomized into four treatment groups ( n = 32/group) and fed diets containing vitamin K 3 (as menadione nicotinamide bisulfite) at 3 (control), 12.9, 23.7, and 45.7 mg/kg feed. Vitamin K 1 , menaquinone (MK)-4, MK-7, and MK-9 were measured in raw whole eggs via a liquid chromatography tandem mass spectrometry method. MK-4 was the most abundant form of vitamin K (91-98%) found in all eggs. Increasing the vitamin K 3 content of hen feed over the control level significantly ( p < 0.001) enhanced the MK-4 content of eggs (mean range: 46-51 µg/100 g, representing ~42-56% of US Adequate Intake values). Vitamin K biofortification also led to significant ( p < 0.05) increases in the yellowness of egg yolk and in eggshell weight and thickness, but no other changes in egg quality or hen performance parameters. In conclusion, high-quality vitamin K-biofortified eggs can be produced with at least double the total vitamin K content compared to that in commercially available eggs.

Background: Red meat and meat products can contribute meaningfully to the mean daily intake of vitamin D. Beef and lamb can contain vitamin D 3 and 25-hydroxyvitamin D 3 [25(OH)D 3 ] but also potentially vitamin D 2 and 25-hydroxyvitamin D 2 [25(OH)D 2 ], all of which contribute to meat's vitamin D activity., Objectives: We aimed to measure the vitamin D 3 , vitamin D 2 , 25(OH)D 3 , and 25(OH)D 2 content of Irish beef and lamb., Methods: Full striploin steaks (longissimus dorsi) ( n  = 39) from beef cattle slaughtered in winter, spring, summer, and autumn as well as lamb steaks (hind leg) from sheep slaughtered in autumn ( n  = 8) were sourced and homogenized. The contents of all 4 vitamin D-related compounds were analyzed using an LC-tandem MS method in conjunction with the National Institute of Standards and Technology's standard reference material no. 1546a-Meat Homogenate. The total vitamin D activity of meat was defined as: {vitamin D 3 + [25(OH)D 3  × 5] + vitamin D 2 + [25(OH)D 2  × 5]}., Results: The median (IQR) total vitamin D activity of striploin beef steak ( n  = 39, irrespective of season) was 0.56 (0.37-0.91) μg/100 g. The content of all 4 vitamin D compounds in beef steak varied significantly ( P  < 0.0001) with season ( n  = 8-11/season group). Median total vitamin D activity of beef steak increased in a stepwise manner ( P  < 0.0001) from winter to the following autumn (increasing from 0.31 to 1.07 μg/100 g). The mean total vitamin D activity of lamb samples ( n  = 8) from autumn was 0.47 μg/100 g., Conclusions: About one-third of the total vitamin D activity of Irish beef was attributable to its combined vitamin D 2 and 25(OH)D 2 content, estimates of which are largely or completely missed in food composition tables. There was significant seasonal variation in all 4 vitamin D compounds as well as in total vitamin D activity, which has implications for vitamin D nutrient claims for beef., (Copyright © The Author(s) on behalf of the American Society for Nutrition 2020.)

Background: In women with prolactinoma medical treatment with dopamine agonists (DA) can restore fertility. A number of studies have established the safety of DA during pregnancy and the impact of pregnancy and lactation on remission of prolactinoma. However, the total number of reported cases remains modest and further evidence is needed., Aims: To evaluate the safety of DA during pregnancy and remission of prolactinoma after pregnancy and lactation., Materials and Methods: Retrospective cohort study (2002-2014) of 57 pregnancies in 47 women with prolactinoma who received DA. Neonatal and pregnancy complications were recorded. Prolactin levels and treatment data were collected at the time of diagnosis, pre-conception, during pregnancy and lactation, and post-partum (up to 114 months)., Results: DA treatment was stopped a median of 4.5 weeks after conception in 49 pregnancies (86%). There were 49 live births (86% of pregnancies) and six miscarriages. Six pregnancies had an adverse neonatal outcome including two with congenital malformations. Following 26% of pregnancies women achieved remission after birth or lactation, and 25% of women were in remission at last follow-up. Remission was associated with older maternal age (P = 0.036), a lower prolactin level at diagnosis (P = 0.037), and a smaller adenoma at diagnosis (P = 0.045)., Conclusions: Successful pregnancy and lactation is common after DA treatment for prolactinoma. Fetal exposure in the first four weeks of pregnancy appears to be generally safe. Encouragingly, post-partum and after lactation a quarter of women had a normal prolactin level without medical treatment., (© 2019 The Royal Australian and New Zealand College of Obstetricians and Gynaecologists.)

Background: The aetiology of pelvic floor dysfunction (PFD) is still poorly understood. However childbearing is recognized as a major risk factor., Objectives: To elucidate the natural history of PFD by investigating the impact of the mode of delivery on postnatal pelvic floor dysfunction in primiparas, when PFD existing before the first pregnancy is taken into consideration., Study Design: 4P-study (Prevalence and Predictors of Pelvic floor dysfunction in Primips) is a prospective cohort study, nested within the Screening for Pregnancy Endpoints (SCOPE) study set in a tertiary referral teaching hospital with 9000 deliveries annually. Established and proposed risk factors for urinary, fecal, prolapse and sexual dysfunction and the severity of symptoms for each of these outcomes were assessed using the Australian Pelvic Floor Questionnaire in 1482 nulliparous women, who each completed the questionnaire in early pregnancy. Of these, 1060 (72%) repeated the questionnaire 12 months postpartum.Outcomes were analyzed using multivariate ordinal logistic regression., Results: Significant (p<0.05) risk factors for postpartum PFD were pre-pregnancy presence of similar symptoms Odds Ratio (OR) (5.0-30.0), smoking (OR 2.2-4.6), recurrent UTI (OR 2.2-17.3), high hip circumference (OR1.4-1.6), vigorous exercising (OR 3.1-17.9), induction of labor (OR 1.5-2.3), forceps delivery (OR 1.8-8.8), and 3rd degree perineal tear (OR 2.4-2.7). Cesarean section was associated with a lower risk of stress urinary incontinence (OR 0.3-0.5). Other common pre-pregnancy significant (p<0.05) risk factors for various PFD types prior to the first pregnancy were: diagnosed depression - (OR 1.6-2.1), high BMI (OR 3.1), strenuous exercising (OR 1.3-2.2), recurrent UTI (OR 1.5-2.5) and lower educational achievement (OR 1.5-1.6)., Conclusions: Pre-pregnancy PFD was mainly associated with modifiable risk factors such as smoking and exercising. The main risk factor for postpartum PFD was the presence of similar symptoms prior to pregnancy, followed by anthropometric and intrapartum factors. Hip circumference seems to be a better predictor of PFD compared to BMI. When pre-pregnancy PFD was included in the analysis, Cesarean section was protective only for stress urinary incontinence, while delivery by forceps increased the risk of prolapse., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Gamma(2)-melanocyte-stimulating hormone (γ2MSH) is a peptide hormone released by the pituitary gland which is thought to act directly on the renal inner medulla to promote increased sodium excretion into urine (natriuresis). The aim of this study was to determine if a stable analog, [Nle(3), D-Phe(6)]-γ2MSH (NDP-γ2MSH), of the native peptide regulated the activity, expression and cellular localization of epithelial sodium channel (ENaC) in a murine inner medullary collecting duct (mIMCD-3) cell line. Our results indicate that expression of the γ2MSH receptor, melanocortin receptor 3 receptor (MC3R), is up-regulated by culturing the cells in media with an increased osmolality (∼400mOsm/kg). Furthermore, stimulation of cAMP signaling and sodium transport by 1nM NDP-γ2MSH occurs only in cells cultured in the high osmolality media. Finally, treatment of mIMCD-3 cells cultured in high osmolality medium for 1h with 1nM NDP-γ2MSH causes a reduction in expression of serum- and glucocorticoid-induced kinase (sgk1) and a reduction in expression and cell surface abundance of the alpha subunit of ENaC. Collectively, this data suggest that γ2MSH directly regulates both ENaC expression and cellular localization in the inner medulla to exert its natriuretic effect., (Copyright © 2013 Elsevier Inc. All rights reserved.)

2, 3, 5, 6-Tetrachloro-1, 4-benzoquinone (TCBQ) is a metabolite of pentachlorophenol known to react with cysteines of glutathione transferases (GSTs). TCBQ treatment of rat kidney rGSTA1-2 and rGSTA1-1 abolishes 70-80% conjugation of glutathione (GSH) to 1-chloro-2, 4-dinitrobenzene and results in strongly correlated quenching of intrinsic fluorescence of Trp-20 (R>0.96). rGSTA2-2 is only inhibited by 25%. Approximately 70% (rGSTA1-1) and 60% (rGSTA1-2) conjugation activity is abolished at TCBQ: GST stoichiometries near 1:1. The inactivation follows a Kitz/Wilson model with K(D) of 4.77+/-2.5microM for TCBQ and k(3) for inactivation of 0.036+/-0.01min(-1). A single tryptic peptide labelled with TCBQ was isolated from kidney rGSTA1-2 containing Cys-17 which we identify as the site of modification. Treatment with more than stoichiometric amounts of TCBQ modified other residues but resulted in only modest further inhibition of catalysis. We interpret these findings in terms of localised steric effects on the relatively rigid alpha-helix 1 adjacent to the catalytic site of subunit 1 possibly affecting the Alpha class-specific alpha-helix 9 which acts as a "lid" on the hydrophobic part of the active site. Homology modelling of rGSTA1-1 modified at Cys-17 of one subunit revealed only modest structural perturbations in the second subunit and tends to exclude global structural effects.

Introduction: Pregnancy and lactation-associated osteoporosis (PLO) is an uncommon condition characterized by the occurrence of fracture(s) during late pregnancy or the puerperium. The aetiology is uncertain, and its management and natural history poorly defined., Methods: We report a series of 11 women with PLO seen at our institution over the past 20 years, with follow-up ranging from 1 to 19 years., Results: Ten women presented with painful low-trauma vertebral fractures, at a median of 1 month postpartum. In nine cases the fractures were multiple (median: 3, range: 2-5). At least one recognised risk factor for osteoporosis (low body weight, smoking history, family history of osteoporosis/fracture, vitamin D insufficiency) was present in nine patients. Bone density was in the osteoporotic range at the spine (mean T score: -2.8), with less marked reduction at the proximal femur (mean T score: -1.9). Nine patients received bisphosphonate treatment, for a median duration of 24 months. In the five women who received a bisphosphonate within 1 year of presentation, spinal bone density increased by 23% over baseline values after 2 years of treatment (p=0.0014). Of the 5 women who had subsequent pregnancies, one, who had declined bisphosphonate therapy after the initial presentation, sustained a fracture in the postpartum period. Two patients (both of whom were followed for at least 10 years) sustained fractures outside of pregnancy., Conclusions: PLO is therefore associated with significant morbidity, a high prevalence of recognized risk factors for osteoporosis and a risk of recurrence in subsequent pregnancies. Bisphosphonate therapy administered soon after presentation substantially increases spinal bone density in patients with PLO.

Female voiding dysfunction is poorly understood; it lacks standard definitions, and there is no consensus on diagnostic criteria. In the majority of women who are neurologically intact the cause is idiopathic. It affects the sufferers' quality of life, but unfortunately there is a paucity of published literature on its management. This review examines the current knowledge on the management of this common problem. Diagnosis is aimed at identifying the underlying aetiological factors, which are discussed, as well as the importance of a detailed history and focused physical examination. Investigations essential to management are outlined. Developments in the medical treatment of voiding dysfunction have been disappointing. The role of surgery is even more limited except for those with postoperative voiding problems after new-generation sling procedures. Intermittent self-catheterisation, supervised and supported by a dedicated nursing specialist, remains the mainstay of management. A multidisciplinary approach is essential to success. Emerging treatment modalities such as sacral and peripheral neuromodulation and the use of alpha(1)-blockers are discussed. Botulinum toxin A injections have been useful in some cases. There are relatively few publications on the effectiveness of these interventions in clinical practice. These issues need to be addressed by quality research. Female voiding dysfunction presents a challenge to urogynaecologists and urologists alike.

Most fungal glutathione transferases (GSTs) do not fit easily into any of the previously characterised classes by immunological, sequence or catalytic criteria. In contrast to the paucity of studies on GSTs cloned or isolated from fungal sources, a screen of databases revealed 67 GST-like sequences from 21 fungal species. Comparison by multiple sequence alignment generated a dendrogram revealing five clusters of GST-like proteins designated clusters 1, 2, EFIBgamma, Ure2p and MAK16, the last three of which have previously been related to the GST superfamily. Surprisingly, a relatively small number of fungal GSTs belong to mainstream classes and the previously-described fungal Gamma class is not widespread in the 21 species studied. Representative crystal structures are available for the EFIBgamma and Ure2p classes and the domain structures of representative sequences are compared with these. In addition, there are some "orphan" sequences that do not fit into any previously-described class, but show similarity to genes implicated in fungal biosynthetic gene clusters. We suggest that GST-like sequences are widespread in fungi, participating in a wide range of functions. They probably evolved by a process similar to domain "shuffling".

Despite advances in the understanding of the pathophysiology of asthma and the availability of effective treatment, the World Health Organization estimates that asthma accounts for 1 in every 250 deaths worldwide. Viruses are associated with half of all asthma exacerbations. The immune response to viral infection may enhance preexisting airway inflammation via the release of chemokines and cytokines and local recruitment of inflammatory cells. Murine models have provided evidence for a deleterious role for CD8+ T cells in the context of respiratory viral infection. Passive transfer of respiratory syncytial virus-specific cytotoxic T lymphocytes (CTLs) to infected mice results in virus clearance, which is associated with acute and sometimes fatal pulmonary disease. Compared with control subjects, CD8+ cells appear to be preferentially sequestered in the airways of individuals with asthma during acute exacerbations. In addition, an expanded CD8+ T cell population, dominated by activated cytotoxic CD8+ lymphocytes, has been documented in biopsies from patients dying as a result of acute asthma in association with a viral infection. Undoubtedly, CD8+ CTLs are a crucial in cell-mediated immunity in response to respiratory viruses. However, it would appear that an aberrant CD8+ T cell response in the context of a viral infection may place individuals with asthma at risk for fatal asthma.

Nuts are high in fat but have a fatty acid profile that may be beneficial in relation to risk of coronary heart disease. Nuts also contain other potentially cardioprotective constituents including phytosterols, tocopherols and squalene. In the present study, the total oil content, peroxide value, composition of fatty acids, tocopherols, phytosterols and squalene content were determined in the oil extracted from freshly ground walnuts, almonds, peanuts, hazelnuts and the macadamia nut. The total oil content of the nuts ranged from 37.9 to 59.2%, while the peroxide values ranged from 0.19 to 0.43 meq O2/kg oil. The main monounsaturated fatty acid was oleic acid (C18:1) with substantial levels of palmitoleic acid (C16:1) present in the macadamia nut. The main polyunsaturated fatty acids present were linoleic acid (C18:2) and linolenic acid (C18:3). alpha-Tocopherol was the most prevalent tocopherol except in walnuts. The levels of squalene detected ranged from 9.4 to 186.4 microg/g. beta-Sitosterol was the most abundant sterol, ranging in concentration from 991.2 to 2071.7 microg/g oil. Campesterol and stigmasterol were also present in significant concentrations. Our data indicate that all five nuts are a good source of monounsaturated fatty acid, tocopherols, squalene and phytosterols.

Epidemiological evidence suggests that dietary intake of carotenoids and tocopherols may influence the risk of certain chronic diseases, such as cancer and CVD. In vitro studies investigating the synergistic effects of mixtures of carotenoids and tocopherols have been hindered due to the difficulty of solubilising these lipophilic compounds. The objective of the present study was to develop a system for delivering tocopherols and carotenoids simultaneously to cells in culture. Differentiated human colonic adenocarcinoma cells (CaCo-2) were incubated with a mixture of these phytochemicals for 24 h. The phytochemical mixture included carotenoids (astaxanthin, canthaxanthin, lutein, lycopene, alpha-carotene, beta-carotene) and tocopherols (alpha-tocopherol and gamma-tocopherol). The emulsifiers polyoxyethylene sorbitan monopalmitate (Tween 40) and polyoxyethylene sorbitan monooleate (Tween 80) were employed as the delivery vehicles, and were compared with tetrahydrofuran (THF). Each vehicle was added at a maximum concentration of 1 ml/l. No toxic effects to the CaCo-2 cells were noted when Tween 40 or Tween 80 were used. Both Tween 40 and Tween 80 resulted in greater solubility of the mixture and delivered substantially more carotenoids and tocopherols to the cells than THF. In particular, lycopene was detected within the cells when Tween 40 and Tween 80 were employed, whereas it was below the limits of detection by HPLC when THF was used as the delivery vehicle. The phytochemicals were retained within the cells for 24 h after supplementation. Tween 40 and Tween 80 have potential as simple, rapid and non-toxic methods for delivering mixtures of carotenoids and tocopherols to cells in culture.

This case report demonstrates Mineral Trioxide Aggregate obturation of the root canal system of a retained primary mandibular second molar where no succedaneous permanent tooth was present. The technique seemed to provide a biocompatible seal of the root canal system in this case. It is not recommended for obturation of primary teeth that are expected to exfoliate since it is anticipated that Mineral Trioxide Aggregate would be absorbed slowly, if at all.

A two-step strategy involving DEAE-cellulose and POROS PI anion exchange chromatography has been developed for rapid purification of acetolactate decarboxylase (ALD) from Leuconostoc lactis NCW1. This results in 5333-fold purification with a yield of 30%. Purified ALD is a dimer of 49-kDa subunits, has a pH optimum of 6.0, a pI of 4.2 and its activity is independent of metals or branched chain amino acids. At the optimum pH, the K(m) for 2-acetolactate (ALA) was found to be 1.3 mM and the turnover number was 4000 min(-1). N-terminal sequence comparison with other ALDs showed little sequence conservation in this region. Purified ALD does not catalyse direct production of diacetyl from ALA, unlike the crude extract.