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Bisphenol A (BPA) is a sealant and flux of plastic materials and has been determined to be an endocrine-disrupting chemical. Prenatal exposure to BPA can lead to substantial adverse effects on fetal growth and development. This study was conducted to assess BPA concentration in pregnant women and umbilical cord blood, and to investigate whether maternal BPA exposure affected fetal outcomes including lower birth weight (LBW), smaller size for gestational age (SGA), and high leptin (HLP) and low adiponectin (LAD) secretion.
We measured the BPA levels of maternal blood (n = 97) and umbilical cord blood (n = 97) with a high-performance liquid chromatography/UV detector. The protein secretion of leptin and adiponectin were separately determined using enzyme-linked immunosorbent assay. A logistic regression was performed to estimate the effects of maternal exposure to BPA on LBW, SGA, and adverse action of adipokines in newborns.
The geometric means of BPA concentration in maternal blood and fetal cord blood were 2.5 ng/ml and 0.5 ng/ml, respectively. Elevated risks of LBW (OR 2.42, 95% confidence interval (CI) 1.72-3.36), SGA (OR 2.01, 95% CI 1.39-3.01), and adverse action of leptin (OR 1.67, 95% CI 1.12-2.25) and adiponectin (OR 1.25, 95% CI 1.52-3.97) were observed in male neonates in the highest quartile of maternal BPA exposure.
Elevated prenatal BPA exposure increased the risk of LBW, SGA, and adverse actions of adipokines in neonates, especially in male infants. These results provide further evidence that maternal exposure is correlated with adverse birth outcomes.
Bisphenol A (2, 2-bis (4-hydroxyphenol) propane; BPA), a chemical compound found in plastic products, is being used increasingly in industrial manufacturing materials. Numerous reports state that BPA production was 2, 214, 000 metric tons worldwide per year in 2003 [1], and 3, 200, 000 tons in 2005 [2]. Because BPA is used to manufacture polycarbonate plastic, epoxy resins and certain dental sealants [3], humans are frequently exposed to BPA released from plastics and food cans in daily life [4]. Therefore, through these daily exposures BPA potentially affects human health.
Previous studies through analyses of BPA in the serum of pregnant women and in cord blood collected at birth have indicated that BPA accumulates early in fetuses [5,6]. Schonfelder et al. [7] stated that BPA levels vary: from 0.3 to 18.9 ng/ml (median 3.1 ng/ml) in maternal plasma, from 0.2 to 9.2 ng/ml (median 2.3 ng/ml) in fetal plasma, and from 1.0 to 104.9 ng/g (median 12.7 ng/ml) in the placenta. BPA can readily cross the placenta [8], and some in vivo experiments have demonstrated that BPA can cause adverse birth outcomes in offspring. Pregnant rats were orally administered BPA at a dose of 10 mg/kg/day resulting in a decreased number of neonates and decreased survival rate [9]. Female genital tracts developed abnormally when the dams were exposed to BPA through gavage [10] or osmotic pumps [11]. Pregnant rats exposed to a low dose of BPA (1 mg/L) in drinking water showed increased adipogenesis in females at weaning [12].
In utero or neonatal exposure to BPA can alter offspring phenotype by stably altering the epigenome, an effect that can be counteracted by maternal dietary supplements [13]. Male offspring were orally exposed at lower dosages of BPA (0.05 mg/kg/day), which led to a significant decrease in weight gain; however, offspring exposed to BPA treatment at a higher dosage (50 mg/kg/day) showed higher body weights than the controls [14].
Mechanistically, environmental BPA is a well-known endocrine-disrupting chemical that binds to estrogen receptors (ER) α and ERβ and results in competition with estrogen [15], while disrupting the folding, assembly, and shedding of many cellular proteins by targeting protein disulfide isomerase [16]. BPA can accumulate in adipose tissue. Fernandez et al. [17] detected an average 3.2 ng BPA/g fat and 8.2 ng chlorinated BPA/g fat in female adipose tissue. Olea et al. [18] found substantially higher levels in children. Moreover, BPA present in adipose tissue may alter the release of adiponectin and leptin, which could influence insulin resistance and increase susceptibility to obesity-associated diseases [19]. Additionally, BPA triggers adipocyte differentiation [20]. The treatment of BPA in mouse 3T3-L1 embryonic fibroblasts stimulates triglycerol accumulation in adipocytes and hepatocytes, and promotes preadipocytes to differentiate into mature adipocytes. The 3T3-L1 cells treated with BPA increase the levels of lipoprotein lipase and adipocyte-specific fatty acid binding protein (aP2) mRNAs. Therefore, BPA might mimic estrogen properties and alter adipokine release, thereby affecting fetal growth. Thus, this study examined the BPA distribution in maternal and umbilical cord blood, and its profile of adipokine protein. We also investigated the effects of prenatal BPA exposure on fetal birth outcomes including fetal development and hormone regulation.
One hundred and fifty-seven healthy pregnant women were recruited for this study between January 2006 and August 2007 at an obstetrics and gynecology clinic in Hsinchu County, Taiwan. All pregnant women consented to participate in this study, and the bio-sampling process was approved by the institutional review boards of National Tsing Hua University. One hundred and thirty-four subjects (response rate 85.3%) completed a self-reported questionnaire including physical characteristics (age, height, weight, occupation, cigarette smoking, betel quid chewing, and alcohol consumption), dietary habit, and disease history. Of the 97 mother-newborn pairs enrolled in the study (72.4%). 37 dropped out before delivery.
Maternal blood corresponding to umbilical cord blood samples were respectively collected in glass heparin tubes at full-term delivery. Plastics were excluded throughout the entire analytic procedure to avoid BPA contamination. Whole blood was centrifuged at 12, 000 rpm for 10 min to separate the plasma and corpuscles, and stored at -80°C until analysis. To the plasma fraction (500 μl) was added 100 μl of 0.01 M ammonium acetate buffer (pH 4.5; Riedel-de Haen, Seelze, Germany) and 4 ml mixture of n-hexane (HPLC grade; Echo Chemical, Miaoli, Taiwan) and diethyl ether (70:30 v/v, anhydrous; J.T. Baker, Phillipsburg, NJ). The samples were mixed for 5 seconds, vortexed for 10 minutes, immobilized for one minute, and then 8.71 μl of 9.187 M perchloric acid (purity 60-62%; Sigma-Aldrich, St. Louis, MO) was added. After centrifugation at 3, 000 rpm for 5 minutes, the organic layer was evaporated to dryness, and reconstituted with 100 μl of mobile phase (methanol:water 80:20 v/v) for BPA determination by a reverse-phase high performance liquid chromatography (HPLC).
The BPA concentrations in plasma were determined using HPLC chromatography (D-7000) connected to a UV detector (L-7400) consisting of an autosampler (L-7200), a pump (L-2130) and a degasys (DG-2410) (Hitachi High Technologies America, Pleasanton, CA). HPLC conditions were as follows: 20 μl injection volume, Inertsil OctaDecylSilane (ODS)-3V column (5 μm, 250 mm × 4.6 mm; GL Sciences, Torrance, CA) fitted with a Metaguard Polaris C18 cartridge (5 μm, 4.6 mm; Varian, Lake Forest, CA) at room temperature, and mobile phase methanol:water (80:20 v/v) with a flow rate of 0.7 ml/min for 20-minute run time. The eluted peak of BPA (bis-(4-hydroxyphenyl)-propane, purity > 99%; Sigma-Aldrich, St. Louis, MO) was detected at 226 nm, and chromatographic data were analyzed by D-7000 multi-system software version 4.1. Both the initial standard stock solution, as well as the serial dilutions from the stock solution in methanol for the HPLC study, were 0.5 mg/ml in methanol. Linear calibration curves obtained for BPA ranged from 3.9-250 ng/ml, and the coefficients of the determinations (r2) were ≥ 0.995. The QA/QC materials were prepared from a plasma pool obtained from multiple anonymous pregnant women donors in analysis with standard, reagent blank, and unknown samples. We performed external calibration using the chromatographic responses of seven standard concentrations in their corresponding solvent. The recovery rates of blanks extended from 96-103%. The relative standard deviation (RSD) among triplicate analyses were 1.99-7.53%, and the recovery percentage was 96.1% with an RSD of 7.53%. Concentrations below the limit of detection (LOD) of 0.13 ng/ml [n = 25 (26%)] were given a value of LOD/√2 for statistical analyses [21].
The concentrations of plasma adiponectin and leptin protein were determined using Quantikine® human immunoassay kits (R&D Systems Inc, Minneapolis, MN) according to the operation manual. A 96-well polystyrene microplate was coated with mouse monoclonal antibodies individually against adiponectin or leptin globular domain conjugated to horseradish peroxidase. After incubation, stabilized hydrogen peroxide and chromogen tetramethylbenzidine were added, and the optical density of each well was determined at 450 nm using a VERSAmax microplate reader (Molecular Devices, Sunnyvale, CA). The levels of adiponectin and leptin were deduced from the absorbance value by extrapolation from a standard curve generated in parallel. The minimum detectable dose of adiponectin and leptin is typically less than 0.246 ng/ml and 7.8 pg/ml.
The newborns were assessed as (a) low birth weight (LBW) defined as a newborn's birth weight less than the 10th percentile (< 2, 600 g; n = 20); (b) small for gestational age (SGA) defined as birth weight less than the 10th percentile, compared with the birth weight distribution in the same gestation week and gender according to the data of nationwide newborn's birth weight percentiles by gestational age in Taiwan [22]; (c) high leptin (HLP) defined as the level of leptin more than 90th percentile (> 9.56 ng/ml; n = 25) in cord blood samples; and (d) low adiponectin (LAD) defined as the level of adiponectin less than 10th percentile (< 10.32 μg/ml; n = 22).
To consider potential confounding variables, a multivariable logistic regression was used to model the odds of adverse outcome variables (LBW, SGA, HLP and LAD). Previous studies have suggested that maternal metabolic parameters (HDL, TC, TRG, leptin and adiponectin) are important confounders relevant to birth weight and adipokine levels [23,24]. Thus, other than maternal age, BMI, serum BPA concentration, smoking and socioeconomic status, the maternal metabolic parameters, i.e., HDL, TC and TRG were selected as the independent variables for newborn LBW and SGA outcomes. Moreover, the maternal concentrations of leptin and adiponectin were included as potential confounders for cord blood leptin and adiponectin concentrations.
All statistical analyses were conducted using the SPSS statistical package (SPSS version 18.0, Chicago, IL). For continuous exposure variables, a categorical analysis was performed to compare the difference of study subjects based on BPA exposure tertile of pregnant women for testing dose response. We used Spearman correlation coefficients to assess the correlation of birth outcomes with categories of the BPA exposure level in pregnant women. Statistical differences were analyzed using the Mann-Whitney U test for nominal variables, and Student's t-test or one-way analysis of variance (ANOVA) for continuous variables. Multiple logistic regression was used to evaluate the odds ratio of potential effects on birth outcome attributed to maternal BPA concentration. All statistical significances were determined with a two-tailed p < 0.05.
Figure Figure1A1A and Figure Figure1B1B showed the distributions of BPA levels respectively in the 97 maternal blood samples and the 97 fetal umbilical cord blood samples. A log normal distribution was used to fit both the maternal and fetal BPA concentrations, which display high fitness (r2 = 0.99). Maternal BPA concentrations were highly variable (ranging from 0.3 to 29.4 ng/ml), whereas most fetal BPA levels were < 1 ng/ml (77% of the samples were 0.3 ng/ml). The geometric means (GM) of maternal and fetal BPA levels were 2.5 ng/ml and 0.5 ng/ml, respectively. The GM of maternal BPA levels was five-fold higher than the fetal umbilical cord blood.
Table Table11 presented demographic characteristics, BPA levels, and lipid profiles of the 97 pregnant women and their newborns who participated in this study. The average (± S.D.) age, height, weight and BMI of pregnant women were 28.8 ± 3.7 years, 160.9 ± 5.2 cm, 65.3 ± 11.2 kg, and 25.2 ± 4.0 kg/m2. The average values for serum BPA, adiponectin and leptin were 5.4 ± 6.3 ng/ml, 6.6 ± 4.7 μg/ml, and 17.9 ± 13.9 ng/ml in pregnant women, and 1.1 ± 2.2 ng/ml, 21.3 ± 8.2 μg/ml, and 4.6 ± 3.5 ng/ml in neonates. The neonates average weight and gestational age were 3109.6 ± 319.0 g and 38.9 ± 1.1 weeks, respectively. Gender distribution among neonates was 57.7% male and 42.3% female.
Based on the GM of maternal BPA levels, study subjects were divided into high- and low-BPA groups. The levels of TC and LDL were significantly higher in the low-BPA group than in the high-BPA group. Additionally, no significant differences in TRG, HDL, adiponectin and leptin levels were observed between these two groups. Neonates in the high BPA group were found to have lower birth weights than those in the low BPA group (p = 0.13). Some relevant differences between these two BPA groups were related to the gender of the infants.
To examine whether BPA exposure in pregnant women affected fetal BPA exposure and adverse outcomes, we analyzed the correlation between maternal BPA level, lipid profile, adipokines, birth weight, and gestational age (Table (Table2).2). The maternal BPA levels showed a significantly inverse relationship to BMI (r = -0.48, at 3rd tertile BPA level), TC (r = -0.67, at 2nd tertile BPA level), and LDL (r = -0.61, at 2nd tertile BPA level), but were positively associated with TRG (r = 0.44, at 1st tertile BPA level). Moreover, the level of leptin displayed a significant correlation coefficient with increased BPA levels (r = 0.39 at 1st tertile, r = 0.50 at 2nd tertile, and r = 0.59 at 3rd tertile BPA level). Conversely, maternal BPA exposure had a significant negative association with neonatal BPA level (r = -0.53 at 3rd tertile BPA level) and birth weight (r = -0.24 at total sample and r = -0.39 at 3rd tertile BPA level).
As shown in Table Table3,3, the crude ORs showed that maternal BPA exposure was associated with LBW (OR 2.65, 95% CI 1.43-3.47) and SGA (OR 1.93, 95% CI 1.44-2.56) for male neonates but was not significantly associated with fetal adiponectin and leptin. Further adjustment for BMI, TC, TRG, HDL, LDL and adipokines attenuated this association, but maternal BPA exposure was still significantly related to an increased risk of fetal LBW (OR 2.12, 95% CI 1.05-2.38) and SGA (OR 1.34, 95% CI 1.13-2.83) in males. Additionally, the ORs of fetal adverse outcomes and the maternal BPA level were not observed in female neonates.
The risks of fetal LBW, SGA, LAD, and HLP were plotted by quartiles based on maternal BPA exposure levels in Figure Figure2.2. Among male infants, the multivariable adjusted ORs of LBW of the second (OR 2.75, 95% CI 1.52-4.22) and fourth quartile (OR 2.42, 95% CI 1.72-3.36) of maternal BPA exposure, in contrast to the first quartile of BPA levels, showed that maternal BPA levels were significantly associated with an increased risk of LBW. The adjusted ORs of SGA of the second (OR 0.24, 95% CI 0.17-0.32) and the third quartile (OR 0.44, 95% CI 0.20-0.53) of maternal BPA level were significantly decreased, but increased in the fourth quartile (OR 2.01, 95% CI 0.99-0.62) (Figure (Figure2A).2A). The dose- response curve at SGA in male infants illustrated a U-shaped curve, but showed adverse effects at the highest BPA exposure (4th quartile). Additionally, significantly increased risks of LAD (OR 1.67, 95% CI 1.12-2.25) and HLP (OR 3.03, 95% CI 2.09-4.54) were observed for male neonates at the highest BPA exposure quartile (Figure (Figure2A).2A). In female neonates, BPA levels in the second (OR 2.99, 95% CI 1.47-4.11) and the fourth quartile (OR 2.17, 95% CI 1.19-3.12) were positively correlated with an increased risk of SGA presenting a Z-shaped curve and adverse effects (Figure (Figure2B).2B). Otherwise, the ORs of LAD were significantly decreased in the second (OR 0.21, 95% CI 0.10-0.21) and the fourth exposure quartiles (OR 0.55, 95% CI 0.34-0.56). The ORs of HLP at the second (OR 0.18, 95% CI 0.12-0.18) and third quartile (OR 0.11, 95% CI 0.06-0.11) of maternal BPA exposure were significantly decreased, but increased in the fourth quartile (OR 1.75, 95% CI 1.04-1.85) (Figure (Figure2B).2B). The dose-response curve of HLP in female infants showed a U-shape, but adverse effects were observed at the highest exposure quartile of BPA.
As BPA recently has been clinically shown in human and animals to cross the placental barrier, the potential effects of maternal BPA exposure on prenatal development has become a more focal area of research [10,25]. Previous studies have shown that prenatal BPA exposure in rodents and human is associated with fetal developmental and metabolic adverse effects, for example, increased birth weight [19,26], reduced gestational age [27], and disrupted thyroid function [28]. These findings raise concerns as to the developmental toxicity since the active BPA dose is considerably lower than the reported concentrations in the serum of pregnant women in the U.S. (5.9 ng/ml) and in South Korea (2.7 ng/ml) [29,30]. This study is the first to show the correlation between maternal BPA exposure and birth outcomes in Taiwan. We analyzed BPA levels in blood samples obtained from pregnant women and umbilical cords. The levels of BPA ranged from 0.3 to 29.4 ng/ml (GM 2.5 ng/ml) and 0.3 to 18.5 ng/ml (GM 1.1 ng/ml) in pregnancy and cord serum, respectively. A recent study indicated that approximately 27% of BPA can readily cross a placenta [8] and that maternal BPA exposure transferred to the fetus can cause developmental abnormalities and other adverse health effects in the offspring [10-13]. This study found a 13% transfer of BPA from maternal blood to fetal cord blood. The results indicated that fetuses can be exposed to BPA in utero despite differences in the placental transfer percentage of BPA in a previous study [10]. In male neonates, this study found a significantly negative correlation between fetal birth weight and maternal BPA level. Additionally, the risks of LBW and SGA and higher fetal leptin and adiponectin levels increased with maternal BPA levels in the highest tertile. That prenatal exposure to BPA seems to cause adverse effects on fetal growth and development is an important concern.
The present study revealed several important findings. First of all, we found a correlation between maternal BPA exposure and birth outcomes. Prenatal BPA exposure in the highest quartile (> 7.04 ng/ml) was inversely associated with birth weight, whereas it was not significantly correlated at the lowest and second quartiles of BPA concentrations. Some animal studies supported our finding that the reduction in weight of both male and female offspring is associated with prenatal or postweaning exposure to BPA [12,31]. Kim et al. [32] reported that administration of a high BPA level (300 mg/kg) during the entire gestational period in Sprague-Dawley rats reduced the weight of the fetuses. Maternal exposure in sheep at BPA levels of 30 to 50 ng/ml during days 30 to 90 of gestation resulted in low birth weight in offspring [33]. However, certain studies provided conflicting results, reporting an increased weight in offspring whose mothers were exposed to BPA during gestation [26,34,35]. Moreover, mice embryos cultured at the two-cell stage in 1 nM or 100 μM BPA showed no noted differences in pup weight at birth [36]. However, at the time of weaning on postnatal Day 21, offspring from embryos exposed to either dose of BPA were significantly heavier than the control offspring. Although this study showed that male fetal birth weight was negatively correlated with a high level of maternal BPA exposure, the BPA treatment level of animal models compared to human exposure should be considered. Lee et al. [30] also indicated that maternal BPA levels have more marked effect on male fetuses than female fetuses, but they found a significant positive correlation between maternal serum BPA levels and fetal birth weight.
Estrogen is known to stimulate cell proliferation for growth and development. Troisi et al. [37] indicated that the serum estrogen level of pregnant women is positively correlated with fetus weight, fetus length, and head circumference, but with no existing correlation between the estrogen level of the umbilical cord and fetus weight. Nagata et al. [38] also found a positive association between birth weight and maternal serum estradiol and estriol levels. Aromatase converts testosterone to estradiol [39]. Prenatal testosterone treatment leads to growth retardation and compromised estradiol-positive feedback, as illustrated by Manikkam et al. [40], leading to growth retardation and consequently resulting in low birth weight in sheep. Savabieasfahani et al. [33] reported low birth weight in offspring because of prenatal exposure to BPA and suggested that the effect may be facilitated through a conversion of testosterone to estradiol. In the present study, the prenatal BPA exposure in male fetuses with low birth weight suggested a response to testosterone feedback. Thus, the magnitude of BPA effect on weight may be influenced by subtle changes of hormones in utero.
Second, our data demonstrated that pregnant women exposed to BPA levels greater than 2.51 ng/ml had a higher risk of giving birth to males with LBW (OR 2.12, 95% CI 1.05-2.38) (Table (Table3).3). Moreover, in the male offspring, the lower and highest quartiles of BPA levels conferred a greater risk of LBW than the lowest quartile of BPA level (Figure (Figure2A).2A). This finding produced a non-monotonic or a U-shaped dose-response curve consistent with the previous report [41]. Additionally, male neonates suffered an approximately 34% higher risk of SGA from a maternal BPA level higher than 2.5 ng/ml (Table (Table3).3). A linear dose-dependent response was noted at increased quartiles of BPA levels (Figure (Figure2).2). Conversely, the secretion of leptin and adiponectin was used as a predictor to assess the potential risk of developing metabolic syndrome in newborns. Additionally, a higher relative risk of increased leptin secretion in both male and female infants corresponding to pregnant women exposed to the highest quartile BPA level (Figure (Figure2).2). As mentioned earlier, our results implied that the effects of maternal BPA exposure produced a U-shaped dose and gender dependence on fetal birth outcomes. These results were consistent with several published studies. Differential processing of high dose BPA relative to low dose contributes to the U-shaped, non-monotonic dose-response curve and gender-difference effects [42,43]. Sex- and dose-dependent differences in weight in response to early postnatal exposure to diethylstilbestrol, a synthetic estrogen similar to the BPA structure, have been reported [35]. Prenatal or neonatal exposure to BPA was correlated with adverse effects on fetal growth parameters such as LBW and SGA. Manikkam et al. [40] reported that exposure to endocrine-disrupting compounds in utero caused fetal growth retardation in sheep. Our data suggests that BPA mimics estrogen in its action, and continued exposure to BPA during gestation is likely to have an impact on the fetus' developmental trajectory. In fact, a growing body of evidence, in addition to our data, had considered LBW and SGA to be a potential outcome of fetal exposure to BPA or endocrine-disrupting compounds [33,35].
Thirdly, our data showed that prenatal exposure to BPA may lead to a potential risk of altering metabolic features in the fetus such as increasing adverse secretion of letpin and adiponectin. Phrakonkham et al. [44] suggested that BPA increases gene expression of adipogenic transcription factors in 3T3-L1 preadipocytes. Perinatal BPA exposure is associated with the over-expression of adipocyte hypertrophy and of lipogenic genes in rats [12]. Moreover, BPA was reported to inhibit adiponectin secretion from human adipocyte explants in a dose-dependent manner [45]. An in vivo study suggested that neonatal estrogenization can influence fetal development including fetus weight [28]. These studies provided indirect evidence supporting our observation of BPA altering the secretion of fetal adipokines. However, more information about the adipogenic effect of BPA in fetuses is required for confirmation.
This study has several limitations. First, the level of plasma BPA was only detected at a single time in connection with delivery. Some imprecision exists because BPA exposure is variable over time [46]. Thus, this study assumed that the BPA exposure sources, for example, living environment, consumption habits, and exposure routes, were sustained, and that the exposure level of BPA influencing fetal development and health would be persistent in maternal plasma, though the half-life of BPA is relatively short. Second, the statistical modeling included only maternal age, BMI, and metabolic parameters as control variables to adjust for birth outcomes (LBW and SGA), but parity, a potential confounding factor for birth weight, was not considered. Additionally, it would be reasonable to concede from the results of this study that maternal BPA levels were not significantly correlated with fetal BPA levels. Nishikawa et al. [47] reported that maternal BPA-glucuronide (BPA-GA) may cross through the placenta and deconjugate to BPA in the fetus. Our results suggest that prenatal exposure to BPA might influence fetal development even though we found no significant correlation was present between maternal and fetal BPA levels due to the differences in the drug-metabolizing system of mother and fetus. However, these inferences still require further confirmation.
The BPA levels detectable in cord blood indicated that BPA can be transported to the fetus across the placental barrier. While earlier animal studies have shown a linkage between BPA exposure and adverse fetal effects, this study quantified and illustrated a potential risk in human neonates. However, obtaining more samples from pregnant women and fetuses under varying exposure levels is necessary to reliably assess the risk posed by BPA to humans.
BPA: Bisphenol A; HLP: secretion of high leptin; HPLC: high performance liquid chromatography; LBW: lower birth weight; LAD: secretion of low adiponectin; SGA: small for gestation age; LOD: limit of detection; TC: total cholesterol; TRG: triglyceride; HDL: high density lipoprotein; LDL: low density lipoprotein; BMI: body mass index.
The authors declare that they have no competing interests.
WCC performed the statistical analysis and wrote the manuscript. JLC was involved in processing the raw data and blood sample collection. CFL conducted the determination of blood BPA and adipokines. YCC designed and tested the power and validity of questionnaires, and trained the interviewers. FCS was responsible to contact the study subjects. CYC directed the studies and revised the manuscript. All the authors contributed to the discussion of data and approved the final manuscript.
This study was supported by Taiwan National Science Council grants NSC 97-2314-B-007-002, NSC 98-2314-B-007-002-MY2 and NSC 100-2621-M-007-002.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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text: siegen 20 Report post Posted March 2, 2009 Full Article @ MotorTrend Honda will ramp up production of its new Insight hybrid starting this April, according to reports. The Japanese automaker is reported to have orders for nearly three times its monthly sales target since the vehicle launched on February 6 in Japan. "The response has been overwhelming," Hiroshi Kobayashi, deputy chief operating officer for Honda's domestic operations told Reuters. "We will have to increase production plans." According to Kobayashi, the wait list for a new Honda Insight is up to one month in Japan, but just how much Honda can increase production is likely to be limited by how quickly suppliers can provide parts. What this translates to for the US is yet to be seen. The 2010 Insight goes on sale in America April 22nd (Earth Day), less than two months from now. I can't quite tell for sure if the article indicates that it is just Japanese Domestic Market Insight production that is increasing, or if production will be increasing worldwide or for the US as well. Share this post Link to post Share on other sites


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Item description for The American Religious Experience: A Concise History by Lynn Bridgers...The American Religious Experience offers a short, accessible introduction to American religious history by an award-winning writer. Recognizing the inter-denominational, inter-religious and multi-cultural perspectives that all contribute to the American religious landscape, this book explores the tension between the central, dominant streams of American Christianity and those groups relegated to the periphery. On the edges of the American mainstream we find the histories of groups rooted in visionary traditions, emotionalized forms of religious practice, and ever-expanding ethnic and racial perspectives. The complexity of the religious scene in the United States now, ongoing tensions between identity and diversity, and the many voices that inform American religious practice today grow directly out of the dynamic history that unfolds in these pages.
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List of organizations with .int domain names
|This article's accessibility is in question. The specific issue is: Table legend and visualization. Relevant discussion may be found on the talk page. Information on making articles more accessible can be found at WikiProject Accessibility. (September 2016)
This is a list of organizations with INT domain names, in alphabetical order of the second-level domain name. The list is not comprehensive. As of June 2012, the INT domain consists of 166 subdomain delegations.[1]
These organizations are generally international organizations established by treaty. Some however (such as YMCA) do not meet current restrictions and were grandfathered in from prior acceptance.
List of organizations[edit]
- Table annotations
* Bodies which are not international treaty organizations or (arguably) related to Internet infrastructure, and so do not meet current .INT requirements.
** Domains connected with Internet infrastructure; these are not international intergovernmental organizations.
*** Domains connected with treaties but which may not have the full status of intergovernmental organization (sites for treaties or treaty secretariats).
- ^ "INT Zone File". Retrieved 2012-06-05.


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my early christmas present
Gissa who's over the moon
I am ready to kick some ass and live infront of the computer for awhile, gonna team up with my brother, Rickard, you better get your ass here and join us…
I've got THE BEST gear right now, I am so stoked.
Who is up for a 3 vs 3??? (or just ready to get their ass kicked by me) :p
Fatal1ty Gamer Headset
Thank you JoppE, Linda, Alexandra and Kevin, you are the best!!!


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In an on going effort to better serve this community we have started a new research and development team. This team will assist in the new foundation of our community and our projects. If interested please private message als
[UA] New research and development team
Posted 24 April 2013 - 01:32 AM
Posted 28 April 2013 - 06:50 AM
skynet detected some message in inbox. Maybe my master will be turned on again...
Posted 28 April 2013 - 05:14 PM
It's a Banana!!!!!!!
Hello to you Sir!
How goes it ?
Posted 29 April 2013 - 06:36 AM
not much changed here, eh ?
Looks like my last words were true.....
anyway, I will keep an eye on UA again.
If you need an advice: Cut down everything you do not need. If you think about a thing more then 30 sec. to find a reason, you don't really need it.
Posted 30 April 2013 - 02:05 AM
Quick update:
Our small team has been working hard to start up a organized task board on trello. Few of the things we have addressed are as followed. Feel free to post your feedback here.
List of active gaming communities
List of popular open source games
List of server based games
UA project ideas
UA content revisions
UA Site Development
and much more......
If you wish to join us on Trello please PM me.
Posted 30 April 2013 - 02:41 AM
Do you really think, starting a new "never-ending-planning-discussion" will solve the problem and the community will grow again ?
Here are my thoughts which should make a new and fresh look.
- The Projects Category should be archived unless a new project is available. This will avoid misleading content and will stop the confusion which new community members wil have. Or does anyone know what phpUA was and should be ?
- The UA start page. It was a nice idea but with those outdated content it is terrible. The visitor will not gain anything useful out of it. Remove it and redirect it to the forum. Make a new one if there is NEW and living content
- The forum / IRC should be the place for the community, so focus in those areas
- The wiki. Every content which is not up to date should be remove or marked as old. Every old or even false content will not be useful. It will make the community look "are-they-retarded-this-is-old-and-wrong..."
- Use the forum, talk about things. Make it look that there are ppl around. Make it "active", then new users will come and even start new projects.
just my 2c.
Posted 30 April 2013 - 03:28 AM
Thanks for the reply, and I fully agree! In fact most of these topics have already been addressed. We are taking a new approach to the community development. We are starting up a few standbox projects and discussing what needs to be changed. You're welcome to view our Trello board. Several tasks have been assigned and several motions to archive outdated material are in place. I am accepting all kinds of feedback at this time. Nothing is being changed at this time however discussion and research are being done.
PM me with an email address and I can send you a trello invite to view the board.
Posted 30 April 2013 - 04:54 AM
I hope it did not sound rude, I'm just direct with my opinions and I expect this from others too.
PM sent.
Posted 04 October 2014 - 01:45 AM
I am following UA from cheating-death times. But i am registered yet. anyway, it seems like UA is going down and staff doing nothing. I saw a post today it was about HLGuard source and you guys lost that source. i am sure it happened to CD too. You guys could update CD like sXe Injected. But you did not. CSGO and CS (goldsrc, yey!) still needs a anti cheat! Believe or not, we need.
I Believe in you guys can do an anticheat with new ideas and community can grow up again. And then, you guys will reach thosands of people. In that time you guys can do great projects with new people.
regards, if you guys still alive.
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0 members, 2 guests, 0 anonymous users


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The Deal
Natural lashes are given enhanced length and volume with a set of lash extensions at this salon in Belsize Park.
The Merchant
Caprice Hair and Beauty provides men, women and children with a range of hair and beauty treatments from a cosy Belsize Park location. Beauty treatments include waxing, threading and nail work, while a selection of massages help patrons relax, and microdermabrasion takes care of visages.


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Private Practice Section
Bridging to Best Practice: Translating Evidence to Improve Efficiency, Effectiveness, and Marketability
E-Learning Course
At a time when Evidence-Based Practice is on the rise, are we truly translating the research into superior practice models? How do we bridge the gap between academia and private practice to ask the best clinical questions and to maximize the efficient dissemination of new information? Research shows a significant lag time, up to 10-15 years, for evidence to be integrated into daily clinical practice. As a group and through direct interaction with attendees, we will establish the barriers, benefits and solutions of not only applying a Best Practice model to a clinic but also participating in clinical research. With representatives from various aspects of our profession, we will have an interactive discussion for clinicians across the spectrum and discuss one proposed Best Practice model.
Course Code:LMS-163
Brought to you by:APTA
Available discounts:
- Non Member (until 12/31/2030) $90.00
- Member (until 12/31/2030) $0.00
* Prices in US Dollar


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Charming Edwardian
This Charming Late-Edwardian (circa 1915) property will delight. Step behind the cute picket fence and evergreen front garden, to find this grand residence comprising 4 bedrooms, 2 bath + modern separate rear Studio and DLUG all set on a 470m2 corner allotment. This property was built to last on solid bluestone and slate foundation, and can be enjoyed now in all its charm or updated to a new era. Features include ornate 11 foot high ceilings, stained-glass features, baltic pine floors and traditional fretwork hallway. Comprising a front sitting room/library, equipped with built in bookcases and a decorative open fireplace, 3 generous sized bedrooms (2 with BIR) all with hydronic heating and decorative ceiling roses. The highlight of the home is a wonderful dining/living room which is divided by a fully working two-way ornate fireplace, making this space perfect for entertaining or unwinding after a long day. Down the hall there is a functional spacious kitchen, equipped with two ovens, one IXL cast iron slow cooker and the other a traditional gas cook top and oven. There is also ample storage, dishwasher and a large walk-in pantry. At the rear of the home you will find the main bathroom with oversized shower, brass fittings, separate bath, plus a lovely light-filled laundry area which overlooks the spacious backyard. Outside there is an undercover entertaining deck area perfect for bbq's. The modern self-contained office or additional living area is equipped with kitchenette and en-suite bathroom can also be accessed via the side gate or through the large DLUG. It provides an ideal office or extra family accommodation. Ideally positioned just minutes walk from Geelong Hospital, and a stone's throw away from Garden St cafes and parks. This home would be perfect for families, those wanting to work-from-home, or re-purposed for central Geelong Medical or Professional consulting STCA. FEATURES Hydronic heating, Ornate OFP, 11 foot ceilings, Self-contained Studio, 470m2 approx, period Geelong Red/polychrome brickwork with refurbished tuck-pointing.


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Seminar Host HotelHampton Inn Hendersonville 155 Sugarloaf Road Hendersonville, NCCall 828-697-2333 Code PPH for the room rate of $159.00 plus tax
Chuck Hill - Seminar email@example.com
Rex C. Truell - Director email@example.com
Depart hotel for sundown on the Blue Ridge Parkway We will travel by caravan to Chuck's favorite overlook on the BRP to photograph the sunset. Sunset will occur at approximately 6:55pm. It can get very cold on the Parkway in Oct. Bring a heavy coat.
Chris will walk you through creating colorful action composites using lots of layers and brushes. He will cover everything from lighting to extraction to final processing. These images can be great sellers if you market them correctly. Learn how to add them to your studio's toolbox!
Great lighting is easy to achieve if you know what to look for. This class will break down advanced lighting techniques into straight forward, easy to understand, and simple steps that will give you superb lighting every time. In this class we will cover - What is good exposure and how do you get it? How to achieve great consistent skin tone. Equipment choices. Main light theory. Fill light theory Hair and Kicker light theory. Background/accent light theory Location lighting anytime anywhere. Four location lighting scenarios and how to work with them And much more.
Have you ever seen a beautiful picture of the Milky Way as it stretches across the night sky, hovering above expansive and majestic landscapes? It's not as hard as it seems, and you don't even need a telescope! Join me as we explore the basics of astrophotography (settings, scene selection, light painting, etc.) inside and outside the classroom to learn how to create beautiful nighttime images! We will begin in the classroom teaching the basics of astrophotography and reviewing all of the crucial steps you need to know to create a night sky photo. Then, we will venture out into the night and apply what we learned inside the classroom to create vivid astrophotography landscapes. At the end of the course, you will possess the basic knowledge to create a portrait of the night sky that will wow and impress your friends!
Plan to be on the Road to Randy's studio in Shelby NC by 9am. It will take approximately one hour to travel from the Hampton Inn to McNeilly Photography in Shelby. For those from the Central or Eastern region of North Carolina this will put you closer to home. If you have never been to Randy's studio you are in for a treat. Studio address is 1241 Fallston Road Shelby NC 28150. Lunch will be provided by Bridges BBQ.


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6712 50th Ave
This is a Multi-Family Home.
6712 50th Ave is located in Woodside, a neighborhood in Other.
6712 50th Ave is a 2-floor, 3-unit Multi-Family Home in Queens's Woodside.
The typical wait time for a taxi at 6712 50th Ave is 20 minutes.
As for the demographics around 6712 50th Ave, the median age is 42, 54% of residents are married, and 25% of residents have at least one child. The median individual full-time income is about $37,396 and about 15% of adults here have a bachelor's degree or higher. Registered voters in this building are 100% Democrat.
Regarding weather, the average summer temperature is 77.4°F, while the average winter temperature is 32.9°F. The average annual rainfall is 44.7 inches and the average annual snowfall is 26.9 inches.


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Concerning the microstructure changes that occur at the surface of UO(2) pellets on irradiation to high burn-up
It is shown that in addition to the precipitation of small gas filled pores, there is a pronounced reduction in the grain size at the surface of UO(2) fuel at high burn-up. These microstructure changes were first observed when the local burn-up exceeded 70-80 MWd/kgM. Generally, the change in microstructure does not penetrate more than 200 microns. However, in a HWR fuel irradiated to 75 MWd/kgM a large part of the pellet cross-section was found to have been affected. Temperature predictions for this fuel suggests that it is the restructuring accompanying thermally activated fission gas release at 1100 to 1200 C that limits the distance over which the microstructure changes occur. Apparently, the formation and fission of Pu is not directly responsible for the change in fuel microstructure. The porosity evidently contains most of the fission gas that is lost from the UO(2) lattice in the region where the microstructure changes take place.
Additional information
Bibliographic Reference: Paper presented: Fall Meeting of the European Materials Research Society, Strasbourg (FR), Nov. 27-30, 1991
Availability: Available from (1) as Paper EN 36393 ORA


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HOUSTON - Katherine Mize went to the drive-thru window in her vehicle to get her order at a Starbucks in Houston. The barista, identified in the lawsuit as Brie, handed Mize a 20-ounce cup of coffee. Mize alleges that Brie squeezed the cup when she handed it to her, causing the top to come off. Mize is seeking $200,000 to $1 million for past and future physical pain, mental anguish, physical impairment, medical expenses, loss of earning capacity and physical disfigurement.
Follow Texas News Blog on Facebook


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Another mountain to climb for White
Five years after half of his tongue was lost to cancer, Nick White is about to run up a mountain.
The Wellingtonian will be one of an elite field running up the slopes of Mt Fuji, the highest mountain in Japan, on July 25.
A keen runner, White says he is in the best shape of his life. "Which is pretty freaky."
In 2009, White was diagnosed with aggressive head and neck cancer. He was given long odds - a 50/50 chance he would live for five years.
He would have died were it not for pioneering surgery by Professor Swee Tan at Hutt Hospital.
The right side of his tongue and 34 lymph glands in his neck were removed, then the tongue rebuilt using muscle from his arm.
Intensive radiotherapy and chemotherapy followed.
It was a long and painful road back to health but he has entered the Fuji Summit Run to raise money for Tan's Gillies McIndoe Research Institute.
He wants to raise $1 for every metre of the climb to the 3776m summit and has already raised $2648 through a givealittle page.
The lingering effects of treatment make running slightly more complicated for White than other competitors.
For example, he can't suck liquid through a straw or out of a CamelBak, a water carrying device beloved by runners. Instead, he sticks with traditional water bottles.
"It's hard for me to speak and once I lose a bit of energy it's even harder, that's for sure."
In the run-up to the event - he flies out on July 20 - he has been running in an oxygen mask to simulate the lower oxygen levels the higher he climbs the mountain. It's a tough race with strict time limits. Starting from the city hall of nearby Fujiyoshida at 7am, runners must reach the summit, 21km away, by 11.30am.
"The time limit is really challenging - anything over that and you're disqualified," White says. "If I can get to the summit, great."
The Dominion Post


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TIANJIN (Reuters) - Airbus SE on Wednesday opened its Chinese completion plant for A330 jets, with hopes that an increased presence in the world's fastest growing aviation market would help boost demand for the firm's profitable but ageing wide-body jets.REUTERS
Professor offers students extra credit for examples of 'white privilege'
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One of USC's best options at receiver isn't actually a receiver
USC's coaches thought Texas had sniffed out their play call on the Trojans' final drive of regulation on Saturday. The plan was for Sam Darnold to throw a pass up the middle of the field to Velus Jones Jr., a slot receiver that USC had lined up as a running back in hopes of a favorable matchup....Los Angeles Times - California, national and world news - Los Angeles Times
Kasey Kahne lands 2018 NASCAR ride, signs with Leavine Family Racing
Kasey Kahne will replace Michael McDowell as driver of the Leavine Family Racing No. 95 car. Kasey Kahne is the latest big-name free agent to solidify his 2018 plans, announcing he will join Leavine Family Racing next season to replace Michael McDowell as driver of the No. 95 Chevrolet. The 37-year-old Kahne is losing is spot with Hendrick Motorsports at the end of the season despite having a year remaining on his contract. Kahne's largely underwhelmed during tenure with Hendrick going 102 races between wins before winning the Brickyard 400 at Indianapolis Motor Speedway in July. Kahne is joining a middle-class, single-car team, a dramatic change from the four-car Hendrick team that is regarded as one of NASCAR's best organizations. LFR was founded in 2011, but didn't begin running fulltime until last season and is winless in 139 Monster Energy Cup Series starts. "I want to thank Leavine Family Racing for this opportunity and will work as hard as possible to help them continue improving," Kahne said. "I am ready for this challenge and look forward to making a fresh start." McDowell and LFR have shown marked improvement this season, with McDowell recording 13 finishes of 20th or better -- compared to nine in 2016 -- including a career-best fourth in the July race at Daytona International Speedway. The team currently sits 26th in the owner standing, four spots higher than the year before. Despite the increased performance, team owner Bob Leavine said Kahne gives the team a driver who's shown he can win at NASCAR's top level in addition to someone it can market to potential sponsors. Kahne has won 18 races in a 14-year career and qualified for the playoffs six times. He is currently ranked 15th in the 16-driver playoff field, coming off a 21st-place finish in the postseason opener Sunday at Chicagoland Speedway. "We look at this as an opportunity to go to the next level," Leavine said. "This deals partially with performance, because obviously, Kasey is a playoff-caliber driver. He has a wealth of information being with a larger organization like Hendrick Motorsports, and we think that will help us. We also look at this as an opportunity for marketing to be able to sell sponsorship. "Overall, we think Kasey is a good fit and felt we needed a change. There is absolutely nothing that Michael did, it was just a matter of us having to make some changes to help us move up the ladder and be a factor in the playoffs. I know Kasey will bring things that will help us." Why it matters: Although a high-profile driver, there was uncertainty whether Kahne would find a ride to continue racing in Cup. And while LFR isn't an upper-echelon team, it does provide him a good opportunity to rebound after an uneven five-year stint with Hendrick. Kahne moving out of the spotlight that comes with be a Hendrick driver spotlight and getting the chance to rejuvenate his career unfortunately comes at the expense of McDowell, whose average finish is only 1.6 positions higher than Kahne this season. McDowell's options to remain in Cup are suspect, unless he can find sponsorship to bring to another mid-size team.Sports News, Scores and Fan Opinion Powered by 320 Sports Blogs
Jimmy Kimmel takes on GOPers new health care bill in scathing opening monologue
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Half way through our 14 Night Adriatic Cruise, the ship headed once again to the port in Venice to return 7 night cruisers, reset the ship for new guests, and begin its second 7 day itinerary – which we'd be joining them for. 🙂
This was our chance to see more of Venice, so we headed straight for the St. Mark's Square via water bus to get in the line to tour St. Mark's Basilica early. The "bus" ride was pretty and it was surreal to sit back and look at the views from the water!
The square was already full of its visitors from all over the world. About 25 million people visit Venice each year.
Bryan's research proved handy as he was able to pre-check our backpack so that we could enter the church without delay.The scaffolding (shown below) on the church itself was the only real eye sore here..but the inside of the church was simply stunning (sorry no photos were allowed of the main rooms) .
We found it challenging to navigate from here to the location we were to meet our Free Walking Tour Guide. I mean, the city's layout with islands, canals and alleys is pretty crazy! Eventually we found our meet-up spot.
I had low expectations for the reliability and professionalism of a free walking tour, but as 3 young professionally dressed adults passed out forms, took attendance and divided the group amongst the three of them for the day, I was already blown away. Our tour guide was Andreas ("On-dray-us") and he was a resident there in Venice who was extremely knowledgeable and entertaining! Instead of the packed famous squares (that we would all easily find on our own later anyway) he took us down back alleys and quiet spots showing us the architectural attempts to help the buildings stay upright, various attempts to build more accommodations in the spacially-limited area, interesting bridges, views, and cultural and historical points of interest we'd never realized without him telling us. The Free Walking Tour turned out to be an incredible experience!
We ate lunch in a small square before continuing the last leg of our tour. The main distraction during lunch was this cat who preyed on the pigeons as they flocked to handouts. It was cute enough that I filmed it, but unfortunately when I did he snagged a poor bird! Just my luck. I was so shocked/horrified I quit filming.
But don't worry! The bird actually got away in the end. And I hate to say it but I don't think he learned his lesson.
After lunch we had a bit more of the tour left and we ended at a good place to take the "must do" gondola ride. It was really hot and I honestly wasn't thrilled about sweating on the cute little boat…but we wanted to do it in case it was our only chance to be together in Venice. Thinking he could get away with it since I wasn't making a special thing out of the gondola ride, before I know it Bryan agreed to split a gondola ride with another older couple, dashing any possibility of the gondola being a romantic memory! That's a man for you! I just shook my head and laughed. Despite it becoming a surprise double date gondola ride, it was nice and the couple was truly very fun and relatable – lucky for Bryan haha!
Our gondolier was quite the ladies' man – haha. He called out to and flirted with every girl he saw on the walkways by our route with his italian charm. It was hilarious!
After this we made our way back to the ship where my parents had boarded and we're waiting for us! I was beyond excited to see them and take them on their first cruise ever!
It began to look like a storm was headed for Venice right as we set sail, making the familiar exit from the city unique and dramatic with the stormy sky. 🙂
But we were on to better weather! After a day at sea, we would have an insanely busy four days – with many crazy adventures and stories. Check back here for our next post featuring our time with my parents running the island of Corfu, Greece!


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INDIANAPOLIS, Ind. -- Randy Schwoerer, a veteran of the music entertainment and auto racing business for more than 30 years, has joined the Project Racing Group (Team P.R.G.) as vice president of marketing and communications. Schwoerer will be...
INDIANAPOLIS, Ind. -- Randy Schwoerer, a veteran of the music entertainment and auto racing business for more than 30 years, has joined the Project Racing Group (Team P.R.G.) as vice president of marketing and communications.
Schwoerer will be in charge of sponsorship development, marketing, corporate hospitality and oversee communications for the sophomore FedEx CART Championship team owned by German natives Andreas Leberle and Christian Danner. The team recently announced that driver Luiz Garcia Jr. will be back for his second year with Team P.R.G. and in addition, CART rookie Michael Krumm joins the team as they will expand to a two-car effort for the 2001 season.
"I have known Randy for many years and have always talked to him about working together," said Leberle. "I am grateful that he has decided to hang his hat here with us. Together we can create a first-class business environment and presence in the FedEx CART Championship.
"We have a lot to prove to establish ourselves and with Randy's help we are certainly a step closer."
The native of Minnesota has had several successful years in the music industry including co-managing the popular pop band "The Jets" to Gold and Platinum album sales. He began his racing career in 1990 by managing Indianapolis 500 champion Arie Luyendyk and later worked with CART drivers Derek Daly, Didier Theys, Parker Johnstone and Greg Moore. Schwoerer also worked with three-time Tour De France champion Greg LeMond.
Schwoerer's creativity was instrumental in implementing comprehensive sponsorship campaigns for Brown & Williamson Tobacco's KOOL brand while he was at the Indianapolis-based U.S.D.C. He later had successful runs with Forsythe Racing, the Klipsch Loudspeaker Company and Damon's Restaurants in the Indy Racing Northern Light Series.
"I'm excited to be working with Team P.R.G.," said Schwoerer. "It's my personal goal to help Andreas and Christian to get the proper funding and business structure that will enable Team P.R.G.to be a competitive team in the CART FedEx Championship Series."
Schwoerer lives in Indianapolis with his wife Germaine. He has three children and one grandchild.
-Ken Severson


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Ok..probably not week, just days..
Last night was the second night of power outage. I was damn tired and hungry. Complained as usual. My dad decided to bring us to Riverview Hotel for some food. Ok, all because of me. But my mom does not want and thus they compromised on going to Greenhouse. You know? The place where I had yucky EXPENSIVE crabs? Last night was my second night there. We wait for about more than five minutes to get served. We waited for more than forty five minutes to get my food served. My food is UNCOOKED. I was supposed to change it yet IT IS A FREAKING FORTY FIVE MINUTES! >< Nevermind then. The food was not that nice. My dad cooks a lot better. Then my mom's came the last after like one hour? It was just some prawn in tom yam sauce over kolo meehoon! OMG.. what took so long? AND THE PRAWN WAS UNCOOK! I thought my mom bite a chilli when she her face turned all red, seemingly felt like vomitting and chocking >< THAT WILL BE THE LAST TIME EVER! ><
Ok forget that yucky food incident. It was midnight and I was tired then. >< But I did walk over to the counter and told the waitress off. "We waited for so long. Yet, the chicken is uncook! The prawn is raw!" Then I strode out with my iPod on, walked towards my dad's car leaving my parents and my brother there to settle the bill. BND25 for such food..ewww
Oh yeah..this is not a rant but more on a happy side of things. I was assigned to work in The Mall the other day? On the 21st of January? I met TEK TING! OMG..I never expected to meet her there. Well, I know she did not too. But oh well, it is so nice seeing her there!! I also met Jia Wei and his cousin, Lesley, Serene, another guy who did not came in to talk to me and also whom I forgotten his name (so sorry!)
I do not know why..but just felt recently, there is so much luck in meeting people! I saw Wei Kee in immigration the other day (20th January) and just last two nights, I saw Junaidi. Ok, he saw me! LOL. It had been such a long time that I did not see him! >< I was in shock and a state of blur when he called me. After a few seconds that felt like an hour, I suddenly recalled who he is LOL
Just yesterday afternoon!!! I met Wan Theng and her two sisters who are extremely friendly! OMG, Brunei is really this small that you meet people whom you seldom meet even when you study in the same university together. Nice meeting them! =))
I am sad and furious to have my dear Pat bullied by people. Even if it is not the case, I still feel that she is on the weaker side! Do NOT mess with my darling Pat or I will curse you like I do now! >< Pat, we shall talk soon! =)) I will get to spend my CNY eve with you =D Take care and see you soon! =D


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Niche finding is the base on which you build your whole affiliate marketing business upon.
Getting it wrong will have a devastating effect on your affiliate marketing business that would be hard to recover from.
Put simply, the niche you are in will determine:
- Which affiliate products you choose to promote
- what topics to write about on your affiliate website
- how much traffic you can expect
- How many sales you could potentially make
With that in mind, I will be guiding you through this article on how to find the most profitable niche for your affiliate website and how to find those gem niches.
What is a Money Niche
There is a great distinction between money niches and free niches, as an affiliate marketer, you will be always looking for profitable niches.
But to find those profitable niches you need to be able to analyze any niche and determine yourself if it will be lucrative for your affiliate website.
I myself few years ago fell into the trap of not knowing how to research my niche probably before jumping in and putting a lot of time and money in it.
The result for me was devastating and I ended up closing my affiliate website and losing 1 year of hard work on it.
But after I realized that my niche wasn't a money niche it was too late, and that's why I want to teach how to tell good and bad niches apart.
There are two types of niches, free and money ones.
Free niches are usually those markets where people are looking for articles to read.
The problems they are seeking solutions for are not of high priority for them or not something they are willing to invest in a product to solve it for them.
If a niche has too many articles and less number of products then it means it's a free niche and something you should avoid basing your affiliate marketing website on.
For example, the "weight gain" niche (which was my niche by the way) is not a money niche; people there are looking for recipes and general understanding of the problem.
See what a Google search return for this niche:
The majority of results (and all the top 10) is bunch of forums and blogs that give advices.
Now imagine you trying to rank your website for this main keyword "how to gain weight" (which has 1000s of monthly searches).
Anyone browsing through the top 10 results will read at least two articles and find the answering satisfying for them, which makes it harder for you to sell your affiliate product.
But a "muscle gain" niche which is a subset of the "weight gain" niche is profitable, and later on I will show you that segmentation can help you turn any niche into a money one.
The Importance of Niche Segmentation
Niche segmentation is about dividing the main niche into smaller ones called "micro niches".
Let's for example segment the weight loss niche:
As you can see, each segment is a market in itself with different types of products being sold.
When you segment your niche you get to stay away from intense affiliate competition, and you also can build your affiliate website to be highly relevant and specific to your micro niche.
Entering a micro niche doesn't mean you will limit your profit potential because you entered a smaller market, in contrast, many micro niches have tens of thousands of people in them.
Even micro niches with just couple of thousands (market size) can be very profitable if you picked the right niche segment to target.
Local niches are a great example of micro niches with small market size and yet very profitable.
For example, if you are an affiliate for SEO Company that provides services to a certain city, local people who search for this service will eventually find your affiliate website.
They will most likely buy from you since you are one of the very few websites that offer help.
In the end, if you can segment your base niche into very smaller parts and picked the part that is known to have high buyers number, then there is no reason to ignore that micro niche.
And there will be no reason for you not to create another affiliate website and target another micro niche.
Your work will be less since all your micro niches (subsets of your main niche) will be related somehow.
How to Find a Profitable Niche
When it comes to finding niches in general, people tend to go for something they are familiar with.
And that is a good strategy, because after all you will only succeed if you target a niche you have immense knowledge in.
This way you can establish your authority to the market and gain trust, and trust is what sells in affiliate marketing.
So for your main niche find what you are really good at, may be a hobby of yours or a life experience (like quitting smoking or losing weight or even fishing).
And using that main niche I will show you how to segment it and find out if it is a money niche or not.
First let me show you how to actually segment your niche.
Go to Google and type in your main niche plus a,b,c,d and up to Z:
From what you can see in the above screenshot, Google's auto-complete feature generates a list of micro niches for you.
You need to browse through them all and find which segment you can provide the most value in.
Now let's take for example the "lose weight pills" segment and see if it is profitable or not.
One thing you should know though, profitable niches tend to have multiple products competing among each other.
That's because many vendors have already studied the niche and found that there is a great demand on products there, so many of them created products to satisfy this need.
Our niche's profitability factor will be to look for paid-products while also trying to figure out the market size.
First do a simple Google search for your micro niche to find out if there are any paid-products there:
We can see that there is no shortage of paid-products in this micro niche.
You can also go further and do Amazon search to see the types of products that are offered in the niche (ebooks, software or physical products).
Now the last step is to try and determine how big your micro niche is, because if your micro niche is big then the potential for affiliate sales is also big.
There are many ways to find this out, but for starters you could use YouTube.
When you enter your micro niche keyword and search for videos on YouTube, you get to the video views on these videos.
High video views mean that there are many people in the niche:
As you can see, over 430K views for a video that has been around for just 8 month is an indication of a large market.
And since this is an already segmented market; it is certainly the right niche choice to make.
Final Words
Always try to be creative with your niches and don't just read a list of profitable niches somewhere and blindly start promoting products in that niche.
Every person has his own niche that suite his knowledge and expertise, try to find yours by doing a good niche finding research like I discussed in this article.
Even some affiliate marketing courses will teach you advanced techniques of finding the right niche and some may even give you some tools that make your research a bit easier.