Profiling the expression of fatty acid-binding proteins and fatty acid transporters in mouse microglia and assessing their role in docosahexaenoic acid-d5 uptake.

While the processes governing docosahexaenoic acid (DHA) trafficking across the blood-brain barrier have been elucidated, factors governing DHA uptake into microglia, an essential step for this fatty acid to exert its anti-inflammatory effects, are unknown. This study assessed the mRNA and protein expression of fatty acid-binding proteins (FABPs) and fatty acid transport proteins (FATPs) in mouse BV-2 cells and their mRNA expression in primary mouse microglia. The microglial uptake of DHA-d5, a surrogate of DHA, was assessed by LC-MS/MS following interventions including temperature reduction, silencing of various FABP isoforms, competition with DHA, and metabolic inhibition. It was found that DHA-d5 uptake at 4°C was 39.6% lower than at 37°C, suggesting that microglial uptake of DHA-d5 likely involves passive and/or active uptake mechanisms. Of all FABP and FATP isoforms probed, only FABP3, FABP4, FABP5, FATP1, and FATP4 were expressed at both the mRNA and protein level. Silencing of FABP3, FABP4, and FABP5 resulted in no change in cellular DHA-d5 uptake, nor did concomitant DHA administration or the presence of 0.1% sodium azide/50 mM 2-deoxy-D-glucose. This study is the first to identify the presence of FABPs and FATPs in mouse microglia, albeit these proteins are not involved in the microglial uptake of DHA-d5.

Environmentally adapted bio-oil compounds-derived polyolesters synthesis: Optimization and properties of base fluids.

Non-edible bio-oil derived from lignocellulosic biomass could be used as environmentally friendly lubricant-ester base stock for maritime and road-type transportations. However, the use of crude bio-oil with highly oxygenated compounds required further upgrading to yield ester that mimicked the characteristics of Group V base oil (polyolesters). In this study, bio-oil based polyolesters was produced via esterification using green biopolymer alginate acid catalyst (Al-Alg). The bio-oil compounds used were acetic acid (AcA), propionic acid (PrA) and levulinic acid (LA), while polyols such as neopentyl glycol (NPG), trimethylolpropane (TMP) and pentaerythritol (PE) were used. Optimization studies revealed that NPG-PrA ester gave the best ester purity of 100%, with 95% of diester selectivity under optimum conditions of 15 wt% Al-Alg, 8 h, 6:1 PrA:NPG and 140 °C. The produced polyolesters showed potential lube characteristics with viscosity index of 76, kinematic viscosity of 2.3 mm2 s-1 at 40 °C and oxidative induction time of 15 min at 100 °C. Furthermore, a reusability study of the Al-Alg catalyst indicated high NPG-PrA diester selectivity (above 90%) for 8 consecutive cycles. The physico-chemical properties of spent Al-Alg catalyst were also discussed.

Comparison of Infrared Thermal Detection Systems for mass fever screening in a tropical healthcare setting.

OBJECTIVES: Fever screening systems, such as Infrared Thermal Detection Systems (ITDS), have been used for rapid identification of potential cases during respiratory disease outbreaks for public health management. ITDS detect a difference between the subject and ambient temperature, making deployment in hot climates more challenging. This study, conducted in Singapore, a tropical city, evaluates the accuracy of three different ITDS for fever detection compared with traditional oral thermometry and self-reporting in a clinical setting. STUDY DESIGN: This study is a prospective operational evaluation conducted in the Singapore military on all personnel seeking medical care at a high-volume primary healthcare centre over a one week period in February 2014. METHODS: Three ITDS, the STE Infrared Fever Screening System (IFSS), the Omnisense Sentry MKIII and the handheld Quick Shot Infrared Thermoscope HT-F03B, were evaluated. Temperature measurements were taken outside the healthcare centre, under a sheltered walkway and compared to oral temperature. Subjects were asked if they had fever. RESULTS: There were 430 subjects screened, of whom 34 participants (7.9%) had confirmed fever, determined by oral thermometer measurement. The handheld infrared thermoscope had a very low sensitivity (29.4%), but a high specificity (96.8%). The STE ITDS had a moderate sensitivity (44.1%), but a very high specificity (99.1%). Self-reported fevers showed good sensitivity (88.2%) and specificity (93.9%). The sensitivity of the Omnisense ITDS (89.7%) was the highest among the three methods with good specificity (92.0%). CONCLUSION: The new generation Omnisense ITDS displayed a relatively high sensitivity and specificity for fever. Though it has a lower sensitivity, the old generation STE ITDS system showed a very high specificity. Self-reporting of fever was reliable. The handheld thermograph should not be used as a fever-screening tool under tropical conditions.

Longitudinal growth and health outcomes in nutritionally at-risk children who received long-term nutritional intervention.

BACKGROUND: The benefits of short-term oral nutritional supplementation (ONS) in undernourished children are well-established. The benefits of long-term ONS in promoting longitudinal growth and health in children who are at risk of undernutrition have not been reported previously. METHODS: In this 48-week prospective, single-arm, multicentre trial, 200 Filipino children aged 3-4 years with weight-for-height percentiles from 5th to 25th (WHO Child Growth Standards) were enrolled. Parents received dietary counselling at baseline, and at weeks 4 and 8. Two servings of ONS (450 mL) were consumed daily, providing 450 kcal, 13.5 g protein and micronutrients. Weight, height, dietary intake using 24-h dietary recalls, and physical activity and appetite using the visual analogue scales were assessed at baseline and weeks 4, 8, 16, 24, 32, 40 and 48. The number of sick days for acute illnesses was collected over the study period. RESULTS: At baseline, mean age was 41.2 months with 50% being male. Weight-for-height percentiles showed the greatest increase in the first 4 weeks (12.1 and 12.8 percentiles, respectively, P < 0.0001) and remained significantly higher than baseline (P < 0.0001) but were relatively stable from week 4 onwards. Height-for-age percentiles increased steadily over time and became significantly higher than baseline from week 24 onwards (P < 0.0001). Appetite and physical activity scores at all post-baseline visits improved from baseline (P < 0.0001), and a reduction in the number of sick days from week 16 onwards was also observed (P < 0.0001). Higher parental education level, being male and higher baseline weight-for-height percentiles were significantly associated with higher ponderal and linear growth over time in repeated measures analysis of covariance. CONCLUSIONS: Intervention consisting of initial dietary counselling and continued ONS helped sustain normal growth after a catch-up growth in nutritionally at-risk children.

Insulin resistance is associated with a metabolic profile of altered protein metabolism in Chinese and Asian-Indian men.

AIMS/HYPOTHESIS: Insulin resistance (IR) is associated with obesity, but can also develop in individuals with normal body weight. We employed comprehensive profiling methods to identify metabolic events associated with IR, while controlling for obesity. METHODS: We selected 263 non-obese (BMI approximately 24 kg/m2) Asian-Indian and Chinese men from a large cross-sectional study carried out in Singapore. Individuals taking medication for diabetes or hyperlipidaemia were excluded. Participants were separated into lower and upper tertiles of IR based on HOMA indices of < or =1.06 or > or =1.93, respectively. MS-based metabolic profiling of acylcarnitines, amino acids and organic acids was combined with hormonal and cytokine profiling in all participants. RESULTS: After controlling for BMI, commonly accepted risk factors for IR, including circulating fatty acids and inflammatory cytokines, did not discriminate the upper and lower quartiles of insulin sensitivity in either Asian- Indian or Chinese men. Instead, IR was correlated with increased levels of alanine, proline, valine, leucine/isoleucine, phenylalanine, tyrosine, glutamate/glutamine and ornithine, and a cluster of branched-chain and related amino acids identified by principal components analysis. These changes were not due to increased protein intake by individuals in the upper quartile of IR. Increased abdominal adiposity and leptin, and decreased adiponectin and IGF-binding protein 1 were also correlated with IR. CONCLUSIONS/INTERPRETATION: These findings demonstrate that perturbations in amino acid homeostasis, but not inflammatory markers or NEFAs, are associated with IR in individuals of relatively low body mass.

Longitudinal study of the socio-demographic determinants of changes in body weight and waist circumference in a multi-ethnic Asian population.

OBJECTIVE: To examine the changes in weight and waist circumference of adult Singaporeans between 1998 and 2005-2007, and the associations of these changes with demographic and socio-economic factors. METHODOLOGY: A prospective study, which followed up participants aged 18-69 years from the 1998 National Health Survey. Analysis was performed on data from 2483 individuals (53% of original sample) who returned for follow-up in 2005-2007. Body weight and waist circumference were measured both at baseline and follow-up. Logistic regression was used to examine factors associated with being overweight and obese at baseline. Linear regression was used to examine changes in weight and waist circumference over time. The variables examined were age, gender, ethnicity, marital status, educational level, housing and employment status, smoking, alcohol consumption and sports activities. RESULTS: Mean weight for the population increased over the follow-up period by 1.48 kg (s.d.=4.95) and mean waist circumference increased by 3.32 cm (s.d.=7.92). Cross-sectionally, those who were overweight or obese were more likely to be Malays or Indians, married, homemakers and have lower educational level. Prospectively, individuals who gained the most weight were younger, more likely to be ethnic minority groups and have the lowest body mass index (BMI) at baseline. They also appeared to be of higher socio-economic status (SES) based on housing type. These associations were statistically significant even after adjusting for other variables. CONCLUSION: Obesity prevention should start early in the younger age. Preventive programs need to reach out to Malay and Indian ethnic groups and those with higher SES. These findings should be used in designing messaging of preventive strategies.

Common genetic variability in ESR1 and EGF in relation to endometrial cancer risk and survival.

We investigated common genetic variation in the entire ESR1 and EGF genes in relation to endometrial cancer risk, myometrial invasion and endometrial cancer survival. We genotyped a dense set of single-nucleotide polymorphisms (SNPs) in both genes and selected haplotype tagging SNPs (tagSNPs). The tagSNPs were genotyped in 713 Swedish endometrial cancer cases and 1567 population controls and the results incorporated into logistic regression and Cox proportional hazards models. We found five adjacent tagSNPs covering a region of 15 kb at the 5' end of ESR1 that decreased the endometrial cancer risk. The ESR1 variants did not, however, seem to affect myometrial invasion or endometrial cancer survival. For the EGF gene, no association emerged between common genetic variants and endometrial cancer risk or myometrial invasion, but we found a five-tagSNP region that covered 51 kb at the 5' end of the gene where all five tagSNPs seemed to decrease the risk of dying from endometrial cancer. One of the five tagSNPs in this region was in strong linkage disequilibrium (LD) with the untranslated A61G (rs4444903) EGF variant, earlier shown to be associated with risk for other forms of cancer.

Microfabricated quadrupole ion trap for mass spectrometer applications.

An array of miniaturized cylindrical quadrupole ion traps, with a radius of 20 microm, is fabricated using silicon micromachining using phosphorus doped polysilicon and silicon dioxide for the purpose of creating a mass spectrometer on a chip. We have operated the array for mass-selective ion ejection and mass analysis using Xe ions at a pressure of 10(-4). The scaling rules for the ion trap in relation to operating pressure, voltage, and frequency are examined.

Lymphoma cells selected for resistance against the cytotoxic effect of oxygenated sterols are also resistant to nonsteroidal antiestrogens.

Oxygenated derivatives of cholesterol and related compounds (oxysterols) have long been known to be cytotoxic to many different cell types. The mechanism of this cytotoxic effect is not fully understood. Our laboratory has earlier reported that oxysterol cytotoxicity resembles that of nonsteroidal antiestrogens in some aspects: (i) the cytotoxic action of both types of compounds is blocked by inhibitors of protein or RNA synthesis, and (ii) both classes of compounds bind with high affinity to the microsomal antiestrogen binding site, a protein which may mediate the cytotoxicity of its ligands. We have now extended these studies by developing cell lines which are resistant to the cytotoxic action of oxysterols. Oxysterol-resistant cells were isolated by exposing two murine lymphoma cell lines, K36 and EL4, to incremental concentrations of 7-ketocholestanol. Intriguingly, the resistant cells thus obtained also exhibited considerable resistance to the cytotoxic effects of nonsteroidal antiestrogens such as tamoxifen and clomiphene, having LD50 values which were 10-100-times higher than that of the parental cells. The resistance appeared to be selective for oxysterols and antiestrogens and did not extend to non-specific toxic agents such as azide, ethanol, Triton-X100, or heat. The biochemical basis of the resistance is not clear but is not due to diminished cellular uptake or increased metabolism of the cytotoxic agents or to changes in the antiestrogen binding protein. The availability of the resistant cell lines should facilitate further studies on the mechanism of oxysterol- and antiestrogen-induced cell death.

Inhibitors of protein and RNA synthesis block the cytotoxic effects of non-steroidal antiestrogens.

Non-steroidal antiestrogens such as tamoxifen are known to exert cytotoxic effects against various cell lines in culture. When the antiestrogens are present at sufficiently high concentrations, their cytotoxicity cannot be reversed by estrogens and is demonstrable even with cell lines which lack the estrogen receptor. The mechanism of this cytotoxicity, which is clearly independent of estrogen antagonism, remains unknown. Using two murine cancer cell lines (the K36 leukemia and the EL4 lymphoma cell line), the human breast cancer cell line MCF7, and two non-steroidal antiestrogens (tamoxifen and clomiphene), our laboratory attempted to determine whether the cytotoxic action of non-steroidal antiestrogens was mediated by a mechanism requiring protein or RNA synthesis. In the case of K36 and EL4 cells, inclusion of tamoxifen or clomiphene in the culture medium regularly caused the viable call count to fall below 20-30% of control in 36-48 h. Under these conditions, the addition of inhibitors of protein or RNA synthesis consistently increased viable cell count in a dose-dependent manner. With cultures of K36 cells grown in the presence of 10 microM tamoxifen, for example, the addition of appropriate concentrations of emetine, cycloheximide, puromycin, or actinomycin D increased the percentage of viable cells to 5.0, 2.4, 4.0, and 4.0 times that of control, respectively. Additional experiments revealed that the macromolecular synthesis inhibitors, while effective in inhibiting protein or RNA synthesis to varying degrees, did not affect the cellular uptake of [3H]tamoxifen, suggesting that their ability to protect cells against antiestrogen-induced cell death was not due to an inhibition of cellular uptake of antiestrogens. In the case of MCF7 cells, however, inhibition of protein synthesis did not protect the cells against the cytotoxic effect of tamoxifen. These observations suggest that non-steroidal antiestrogens may exert their cytotoxic effect by at least two different mechanisms; only one of these require de novo protein synthesis. The effect of antiestrogens on K36 and EL4 cells may provide a useful system for the identification of proteins involved in cell death.