5 h of batch fermentation. For the preliminary fed-batch studies, two predetermined feeding profiles, namely exponential and constant feeding were preferred. For each feeding profile, three feeding rates were evaluated: 1, 3 and 6 g

glycerol/L/h for constant feeds and 0.1, 0.2 and 0.3 h−1 for exponential feeds. To achieve the desired rates (1, 3 and 6 g glycerol/L/h), several feed mediums with different glycerol concentrations were prepared. For these assays, the three feeding rates were tested as duplicates (A and B), without induction, so that the growth profiles could be established (Fig. 3). In these fed-batch experiences, glycerol was measured as mentioned in Section 2.2.4 until the end of the feeding SCH772984 process. The growth curves for these PFI-2 solubility dmso profiles (Fig. 3) show a maximum OD of about 50 which, as expected, is considerably higher than those obtained in the batch experiments. For the 1 g/L/h constant feeding profile, glycerol concentration was kept close to zero until the end of the fed-batch process, meaning that these cultures were able to consume all of the glycerol provided by the feeding

solution. For the 3 g/L/h constant feeding profile, glycerol concentration reached close to zero values only after about 10 h of fed-batch, meaning that limiting concentrations are not reached during most of the fed-batch process. However, the maximum OD reached (52) was very similar to that of the 1 g/L/h feeding profile. Finally, for the 6 g/L/h feeding profile, glycerol concentrations either increased throughout the experiment (replicate A) or were kept constant at relatively low levels (replicate B). Since glycerol concentrations during the fed-batch phase of the feeding profiles evaluated were very different (from almost 0 g/L to as high as 30 g/L), cytometry assays were used to see if the feeding profile of 1 g/L/h was, in

fact, the best choice among the three constant feeding profiles tested. In order to assess cell physiology during the fed-batch experiments, flow cytometry assays were carried out using a PI/BOX dual staining. Dead cells will be stained with both BOX and PI, cells with depolarized membrane will be stained only with BOX and viable cells will not be stained. The results (not shown), indicate that as fermentation time increases, the percentage of dead cells (stained with PI and BOX) also increases. This effect is heightened at ADAMTS5 higher feeding rates, possibly because of the higher glycerol concentrations, which can hamper E. coli growth. In fact, at the end of the fermentation, the average percentages of viable cells were 79.43, 65.84 and 75.61% for 1, 3 and 6 g/L/h, respectively. The three chosen specific growth rates for exponential feeding profiles were 0.1, 0.2 and 0.3 h−1 with feed medium addition speed being calculated according to an equation previously described [14]. For this set of experiments, the three specific growth rates were also performed in duplicates (A and B) without induction (Fig. 4).

In find more secondary endosymbiosis, a red alga or a green

alga was engulfed by a non-photosynthetic protist (Green, 2011 and Reyes-Prieto et al., 2007). Chloroplasts of algae belonging to the heterokonts, which include diatoms, brown algae, raphidophytes and heterotrophic oomycetes, arose from a secondary endosymbiosis event including a red alga. Recent results indicate that the red algal endosymbiont succeeded a green algal endosymbiont related to prasinophytes, as a large number of nuclear genes in diatom genomes have a green algal origin (Jiroutová et al., 2010 and Moustafa et al., 2009). However, this finding is controversial, and has been the subject of criticism for taxonomic sampling bias (Burki et al., 2012 and Deschamps and Moreira, 2012). In addition to the large contribution of genetic material to algal genomes through endosymbiosis (endosymbiotic gene transfer, EGT), several genes have been introduced to nuclear and organelle

genomes independently through horizontal gene transfer (HGT) events. The nuclear genomes selleck inhibitor of the diatoms Thalassiosira pseudonana and Phaeodactylum tricornutum contain several hundred genes that appear to have been acquired from a wide range of bacteria through HGT ( Armbrust et al., 2004 and Bowler et al., 2008). Diatoms (Bacillariophyta) constitute one of the most abundant groups of marine phytoplankton, with an estimated diversity of around 100 000 species (Round et al., 1990 and Van den Hoek et al., 1995). The evolutionary success of diatoms is also reflected in their ecological importance; this group contributes approximately 40% to primary net production in the oceans (Field et al., 1998). This success is suggested to be caused at least in part by the ability of diatoms to respond and adapt to large fluctuations in light irradiance, thereby maintaining high photosynthetic efficiency over a wide range

of environmental conditions (Depauw et al., 2012). Thus far, the chloroplast genome has been sequenced in five diatoms: the centrics Roflumilast Odontella sinensis and T. pseudonana, and the pennates P. tricornutum, Fistulifera sp. JPCC DA0580 and Synedra acus ( Galachyants et al., 2012, Kowallik et al., 1995, Oudot-Le Secq et al., 2007 and Tanaka et al., 2011). In addition, the chloroplast genomes of the diatom endosymbiont of two dinoflagellates, Durinskia baltica and Kryptoperidinium foliaceum, have also been characterised ( Imanian et al., 2010). These genomes share a highly similar gene set, of which a core set of 86 genes is found in all chromalveolates ( Green, 2011). Two plasmids identified in the pennate diatom Cylindrotheca fusiformis may be associated with chloroplasts, as they hybridise with chloroplast DNA ( Hildebrand et al., 1992 and Jacobs et al., 1992). In support of this view, genes encoding putative proteins with similarity to ORFs found in the C.

The retrospective design is a significant bias concerning the data collected, which can have an impact on results. Randomized studies are difficult to implement, given the small number of patients, acquisition of one specific technique per referral center, and the fact that the patients are often specifically referred for a selected technique of treatment. However, we think that the consistency of treatment www.selleckchem.com/products/Y-27632.html over the whole study and the detailed

long-term clinical outcome allow us to provide data useful to clinical practice. In conclusion, flexible endoscopy has become a new standard for ZD treatment when performed with adequate equipment inspired by rigid diverticuloscopes. This approach has a lower rate of adverse events than open surgery or endoscopic stapling techniques. Moreover, the risk of general anesthesia in elderly patients can be avoided because airways are protected by the diverticuloscope. Recurrence

is about 25% in the long term but is easily amenable to successful repeated endoscopic treatment. “
“Zenker’s diverticulum (ZD) is an acquired disease that is formed by outpouching of hypopharyngeal mucosa between the inferior pharyngeal constrictor and the cricopharyngeus muscle in an area of junctional muscle weakness known as Killian’s triangle.1 Although the need for myotomy in addition to surgical correction of the diverticulum has been well described, it is only in the past 20 years that more clear insight has emerged on the pathophysiology of ZD despite its original description in the 1700s.1 Specifically, Cook et al2 elegantly demonstrated Navitoclax that in patients with ZD, the upper esophageal sphincter is fibrotic, contributing to reduced compliance, incomplete opening, and therefore increased pressure proximal to the cricopharyngeal Depsipeptide ic50 outlet. This leads to a “blow out” of the weakest part of the pharyngeal wall and formation of the diverticulum. Other studies have been conflicting in demonstrating

an increased tone within the sphincter.1 Nevertheless, these and other data have reinforced that cricopharyngeal myotomy is essential in relieving symptoms and preventing recurrent diverticula.1 The traditional approach to cricopharyngeal myotomy and to diverticulectomy or -pexy has been open through a lateral neck incision. In the past 2 decades, however, this operation has been shifting to a transoral endoscopic approach using a rigid endoscope because of equal efficacy, shorter hospital stay, faster return to oral intake, and lower morbidity because of a reduction in adverse events compared with open surgery.3 Such adverse events may include injury to the recurrent laryngeal nerve, mediastinitis, and fistula.1 Not all patients with ZD are amenable to transoral endoscopic therapy using a rigid endoscope.

For AFB1, its high value of IC50 is different from literature reports measured by MTT

test [10], and this is likely due to different methods used to measure cell viability. SRB refers to the total protein in the cell [22] while MTT test is based on the enzyme activity of NAD(P)H-dependent cellular oxidoreductase [32], so there is possible discrepancy between the two methods, and the value of IC50 is likely dependent see more on the cell viability measurement method. Regarding different values of IC50 between AFB1 and ST, there is a literature report that the IC50 of AFB1 (10 μM) is greater than that of ST (3.7 μM) in human lung cancer cell line of A549 [10] and [33], which also showed that ST is more toxic than AFB1. Another literature report [11] also showed noticeable difference between AFB1 and ST in hormonal induction of tyrosine aminotransferase with different values of IC50 in a rat hepatoma cell line of H4-II-E. The cytotoxicity endpoints of ROS, mitochondria membrane permeability (MMP), DNA

and ATP content all showed cytotoxicity of AFB1 and ST (Fig. 3) to HepG2 cells, and all the endpoints show similar trends when HepG2 cells were exposed to individual AFB1 and ST or their combinations. The contents of both ATP and DNA were decreased while the ROS and MMP were increased p38 MAPK pathway along the treatment concentrations. Comparatively, the decrease of ATP and DNA is more evident than the increase of ROS and MMP, and ST is more potent

to decrease ATP content. However, no significant difference between the measured combinative toxicity and the calculated Histone demethylase toxicity (by adding the values of each endpoint at corresponding concentrations used in their combinations) demonstrates an additive nature of their combinative cytotoxicity. Correlation analysis on the relationship among all the endpoints showed that ATP is positively correlated to DNA content, but negatively correlated to ROS and SRB at a significant level. Both ROS and MMP are positively correlated to SRB (Table 1). Thus, decreased cell viability of HepG2 cells islikely caused by the production of ROS, increased MMP and decreased ATP and DNA when exposed to AFB1 and ST. Consistently, the PCA analysis of these endpoints showed that three clusters can be differentiated: SRB is one cluster, DNA and ATP content is the second cluster, and the third cluster includes ROS and MMP. Considering the biochemical processes associated with mycotoxin exposure, the increased intracellular ROS is a common feature for AFB1 or other mycotoxins [34]. The increased intracellular ROS might cause cross-linking of mitochondria membrane protein and to induce membrane permeability transition and increased MMP [35]. The increase of MMP would result in a decrease of mitochondrial membrane electrochemical potential and uncoupling ATP production from mitochondrial respiratory chain, which would lead to a reduction of ATP production.

Attempting to nurture their young careers has been the ultimate joy of my academic life. “
“Tuberous sclerosis complex (TSC) is a genetic disorder with multi-system involvement that can affect most organ systems.1 and 2

The physical manifestations NLG919 mouse include benign tumours in the heart, kidneys, lungs, skin and brain. TSC is caused by mutations in either of two genes, the TSC1 gene (9q34) 3 and 4 or the TSC2 gene (16p13.3). 5 and 4 TSC has a birth incidence estimated around 1 in 6,000 6, 7 and 8. Appropriate management and coordination of medical specialist care is crucial across the lifespan of individuals with TSC to limit morbidity and mortality in this disease. 9 TSC is also associated with a vast range of neuropsychiatric disorders.10, 11, 12 and 13 At a behavioural level, difficulties include restless and impulsive behaviour, high rates of aggression14, 15, 16 and 18, temper tantrums15 Metabolism inhibitor and

self-injury15, 16, 17 and 18. At the psychiatric level, developmental disorders, including autism spectrum disorders (ASD, 40-50%)19 and attention deficit hyperactivity disorder (ADHD, 30-50%) are commonly seen.10, 12 and 20 High rates of depressive and anxiety disorders have also been reported.15, 20, 22 and 23 At the intellectual level, approximately 50% of individuals with TSC have normal intellectual abilities, and others have varying degrees of intellectual disability.13, 24 and 25 At the academic level, many school-aged children with TSC have academic difficulties, for instance in mathematics, reading writing and spelling.13 At the neuropsychological level a range of neuropsychological deficits are also seen. These include difficulties with executive,

attentional, memory, and language skills.12, 13, 26, 27, 28, 29 and 30 At the psycho-social level, there is growing evidence of the impact of TSC on, for instance, self-esteem, family stress and parental relationships.31 Each individual with TSC will present with their own unique combination of strengths and weaknesses, and this profile may change over time. Taken together, the majority of individuals with TSC will have some neuropsychiatric problems in their lifetime, with lifetime prevalence rates in the region of 90%.32 In 2010 a survey of members Vitamin B12 of the Tuberous Sclerosis Association in the United Kingdom indicated that only 18% of individuals with TSC had ever received an assessment or treatment for neuropsychiatric disorders (personal communication P.J. de Vries). These results suggested a treatment gap of around 70%. At the 2012 International TSC Consensus Conference9 the Neuropsychiatry panel expressed concern about the enormous treatment gap and about the confusion of terminology across different levels of investigation of the bio-psycho-social aspects of TSC.

In women with CD, the rate of new clinically recorded fertility problems was highest in the 25–29 year age group (12.5 per 1000 person-years), and in women without CD

the rate was highest in the 30–34 year age group (12.6 per 1000 person-years). Across all age groups, however, there were no statistically significant differences between the rates of new clinically recorded fertility problems selleck screening library in women with and without CD (eg, IRR in the 25–29 year age group, 1.12; 95% CI, 0.88–1.42; IRR in the 30–34 year age group, 0.87; 95% CI, 0.70–1.08). Of the 290 women who had CD and a recorded fertility problem, 122 (42%) were classified as having undiagnosed CD and 168 (58%) were classified as having diagnosed CD in relation to the new clinically recorded fertility problem. The diagnosis of CD happened at a median of 2 months after the fertility problem (IQR,

4 years before, 2.7 years after). Figure 1 shows the time of new clinically recorded fertility problems in relation to the CD diagnosis. Approximately a quarter of the fertility problems were recorded within a year before or after the CD diagnosis, with 5% being recorded within a year before DAPT research buy the fertility problem and 19% within a year after the fertility problem. Overall, the age-specific rates of new clinically recorded fertility problems were higher in women with diagnosed CD compared with women with undiagnosed

CD (15.4 per 1000 person-years in diagnosed CD compared with 9.8 per 1000 person-years in undiagnosed CD in the 25–29 year age group) (Table 2). There was no statistically significant difference between the rates of new clinically recorded fertility problems in women with both undiagnosed and diagnosed CD compared with women without CD, except for the 25–29 year age group, in which women with diagnosed CD were Ribonucleotide reductase 41% more likely to have new clinically recorded fertility problems compared with women without CD (IRR, 1.41; 95% CI, 1.03–1.92). However, the absolute excess risk was only 0.5% (5.2 per 1000 person-years). Of the 6506 women with celiac disease, 1143 (17.6%) were recorded as symptomatic (with weight loss, diarrhea, or anemia) in the year before diagnosis. The age-specific rates of new clinically recorded fertility problems in this subset of women with symptomatic celiac disease were not statistically significantly different compared with women without celiac disease. The overall rate was found to be 40% lower, however, the absolute risk difference was only 2.3% (Table 2). Of 6506 women with CD, 4649 (71.4%) had received a gluten-free prescription. Of these women, 211(4.5%) had clinically recorded fertility problems, which was almost exactly the same as in the overall population.

This interpretation is consistent with behavioral research suggesting that executive control mechanisms may

be more efficient in bilinguals compared Y-27632 cost to monolinguals (e.g., Treccani, Argyri, Sorace, & Della, 2009). Most relevant to the current study are Blumenfeld and Marian’s (2011) findings that bilinguals’ (but not monolinguals’) inhibition of competing phonological information is associated with the group’s executive control ability. Here, we show that the behavioral differences observed between monolinguals and bilinguals in past research may indeed be driven by differences in how the groups recruit executive control resources at the neural level. Although monolinguals and bilinguals in our study did not differ in their behavioral Simon effect performance (as participants were young adults at their cognitive peak; see Hilchey & Klein, 2011), cortical changes attributed to language experience

emerge even in the absence of behavioral differences between groups (e.g., Bialystok et al., 2013 and Rodríguez-Pujadas et al., 2013). Accordingly, we observed significant correlations between performance on a non-linguistic competition task and cortical activation in regions associated with executive control during a linguistic competition task. Past research has demonstrated that non-linguistic competition is managed through the recruitment of frontal cortical regions including middle frontal gyrus (MFG; Fan et al., selleck screening library 2003 and Maclin et al., 2001), superior frontal gyrus (SFG; Fan et al., 2003 and Maclin et al., 2001), Mirabegron anterior cingulate cortex (ACC; Fan

et al., 2003, Kerns, 2006, MacDonald et al., 2000 and Peterson et al., 2002), and inferior frontal gyrus (IFG; Fan et al., 2003 and Peterson et al., 2002). When faced with linguistic competition, the bilinguals who were best at resolving non-linguistic competition were most likely to strongly activate this extended network of frontal regions. Specifically, correlations between non-linguistic competition resolution and the control of linguistic competition were found in bilateral MFG, bilateral SFG, right IFG, and ACC. This suggests that, in bilinguals, the substrates used to resolve linguistic and non-linguistic competition are highly related. In other words, bilinguals rely on inhibitory control processes that are modality- and task-independent. Monolinguals, in contrast, appear to rely on partially distinct mechanisms for the control of linguistic and non-linguistic competition. Unlike the bilinguals, for whom correlations emerged in multiple distinct regions associated with executive control (bilateral MFG, bilateral SFT, right IFG, ACC), monolinguals’ performance only resulted in significant correlations in right MFG. The finding that bilinguals’, but not monolinguals’, cortical control of linguistic competition is subserved by domain-general control mechanisms is consistent with both neuroimaging (Garbin et al.

Grape cell suspension cultures have been reported to accumulate stilbenes including trans-resveratrol, trans/cis-piceid, ɛ-viniferin, δ-viniferin, pterostilbene, and trans-astringin [9] and [10]. However, the

accumulation of resveratrol in untreated grape cell cultures is low, less than 0.01% of dry weight or 2–3 mg/L [11]. The production of secondary metabolites in plant cell and tissue cultures can be enhanced by elicitors [12]. A number of elicitors including UV, methyl jasmonate, and indanoyl-isoleucine triggered the production of secondary metabolites, including resveratrol [10], [13], [14], [15] and [16]; however, the roles of many other potential elicitors remain to be investigated. If secondary metabolites are click here secreted, in situ adsorption is considered. Amberlite XAD-7, hereafter XAD-7, surpassed other XAD adsorbents in adsorption of many antioxidants SNS-032 purchase including α-tocopherol and α-tocopheryl acetate, which share several common characteristics of resveratrol [17]. In situ adsorption might be crucial, as exogenous resveratrol at a concentration greater than 100 μM or 22.8 mg/L inhibited cell

growth of V. vinifera cv. ‘Pinot Noir’ in a dose-dependent manner [18]. In this study, the elicitation of seven compounds, including jasmonic acid (JA), salicylic acid (SA), 3-methyl-salicyclic acid (MeSA), betaine (BET), β-glucan (GLU), methyl-β-cyclodextrin (MeCD) and chitosan (CHI) was investigated in single and combined treatments for enhancing the production of resveratrol in V. vinifera L. cv. Gamay Fréaux cell suspension cultures. As resveratrol was found secreted into the medium, the elicitation technology was then combined with in situ adsorption and artificial extracellular storage for optimizing resveratrol

production, with a view toward large-scale production. Unless indicated, all chemicals were purchased from Sigma (Australia). The V. vinifera L. cv. Gamay Fréaux cell line was a gift from Dr. Francois Cormier (Québec, Canada). This cell line was grown in GC-2 medium pH 5.7–5.8, which is B5 medium supplemented with 30 g/L P-type ATPase sucrose, 250 mg/L casein hydrolysate, 0.1 mg/L α-naphthaleneacetic acid, and 0.2 mg/L kinetin. Cell suspension cultures were maintained on a reciprocating shaker (Ratek Instruments, Australia) at 100 strokes/min at 27 ± 1 °C. The cultures were kept in the dark to prevent the biosynthesis of anthocyanins that complete with resveratrol and other stilbenes for the same precursors. Pre-cultured 7-day-old cell suspensions were filtered through a 50 μm stainless mesh (Endecotts Ltd. London, England), and the cells were transferred in 20 mL fresh GC-2 medium to reach the concentration of 50 g fresh cells/L. The flasks in triplicate were incubated on a reciprocal shaker (Ratek Instruments, Australia) at 100 strokes/min in a dark, temperature-controlled room at 27 ± 1 °C.

, 2003 and Meng et al., 1999) that led to significantly increased acceptability ratings compared to non-question contexts (Bornkessel & Schlesewsky,

2006b). A set of 160 experimental trials (40 trials per condition) was constructed. Each trial consisted of a three-sentence discourse depicting a scene of two animals performing a transitive action in which both were equally plausible to be the agent or patient of the scene. All trials followed the structure shown in Table 1. (1) In the first sentence (lead-in) of each trial, the current scene with both animals and the instrument of the to-be-performed action was introduced. Thus, in terms of information structure, the relevant characters were discourse-given (Prince, www.selleckchem.com/products/nutlin-3a.html 1981) and the action was inferable (Prince, 1992)

from the instrument mentioned. The same lead-in was used for all conditions. (2) The find more following wh-question (i.e., context question) differed with regard to the factor CONTEXT TYPE: The context question either induced a wide scope of the scene (NEUTRAL CONTEXT) or indicated one of the two animals as the aboutness topic (TOPIC CONTEXT). (3) The third sentence (target sentence) provided a plausible answer to the preceding context question by describing the final action event of the two animals. The target sentence varied according to the factor WORD ORDER and was thus presented in SO or OS order. The different scenes were created based on 40 animals (monomorphemic nouns, masculine gender, 1-syllabic (n = 18) to 2-syllabic (n = 22)) and 10 actions (monomorphemic verbs, transitive, accusative-assigning, 2-syllabic) with corresponding instruments and a scene-setting prepositional phrase (e.g., in the park). Note that both grammatical and thematic roles coincided (i.e., the grammatical subject was always the agent, the grammatical object was always the patient). The critical nouns and verbs were matched for written lemma

frequency, type frequency and normalized log10 familiarity values, taken from the Cyclin-dependent kinase 3 dlex database ( Heister et al., 2011). To control for position effects, each noun occurred once in each of the four conditions at the first and second noun phrase position of the target sentence. Thus, each animal served four times as the agent and four times as the patient of the target sentence, respectively, always with a different action and co-animal. In the lead-in sentence, the first and second mention of the potential agent and patient was counterbalanced across conditions. Both animals of a scene always differed in the initial phoneme. To minimize possible effects of structural priming ( Scheepers & Crocker, 2004), all trials were pseudo-randomized such that maximally two consecutive trials were of the same condition or had the same word order in the target sentence.

“
“Scientific and technological development brings benefits and advantages to our modern lifestyle. Innovation is currently a necessity due to the great demand for new consumer products, but this also brings serious consequences to the current and future generations due to factors such as air, soil and water pollution as related to the release of several chemicals potentially harmful to the environment and human health. Amongst these compounds are the brominated flame retardants (BFRs) that represent a class of contaminants widely used in consumer products due to their high Idelalisib research buy efficiency in inhibiting or minimizing the effects caused by fires, and their low cost; representing 25% of the world market of flame

retardants (Hardy, 1999). However it has been shown that they persist in the environment and show high bioaccumulation potential, Tyrosine Kinase Inhibitor Library being classified as persistent organic pollutants (POPs). Polybrominated

Diphenyl Ethers (PBDEs) are a class of BFRs used as additives in plastics, textiles, electronic circuits and equipments, building materials and many other consumer goods. They are added during the manufacture of various products in daily use, but no effective chemical bonds occurred during the process which would cause their release into the environment during manipulation or improper disposal (Mcdonald, 2002). The bioaccumulation potential of PBDEs and their persistence in the environment are due to their lipophilicity, and high levels of these compounds have been detected in samples of animal fats, blood, placenta and breast milk. (Covaci et al., 2009, Hites, 2004, Li et al., 2008, Ma et al., 2012, Shen et al., 2010, Letcher et al., 2010 and Toms et al., 2007). The HSP90 main contamination routes for humans are house dust and contaminated foods (Branchi et al., 2003 and Talsness, 2008). Amongst the effects described as caused by exposure to PBDEs, there is evidence of a neurotoxic potential (Branchi et al., 2003, Madia et al., 2004 and Verner et al., 2011) and changes in the endocrine system, by acting

on hormone receptors such as estrogen and progesterone, and decreasing the levels of the thyroid hormones (Costa and Giordano, 2007, Costa et al., 2008, Madia et al., 2004, McDonald, 2002 and Zhang et al., 2008). They have also being related to the development of liver toxicity and thyroid cancer (Albina et al., 2010, Hu et al., 2007 and Zhang et al., 2008), but the mechanisms underlying these effects are still not completely understood. 2,2′,4,4′ Tetrabromodiphenyl ether (BDE-47) and 2,2′,4,4′,5 pentabromodiphenyl ether (BDE-99) are the most commonly found congeners in environmental samples and biological systems, and show high levels of toxicity. In vitro investigations have shown that some PBDE congeners, such as BDE-47 and BDE-209, present cytotoxic potential in several cell lines such as HepG2 ( Madia et al., 2004, Jing et al., 2010, Weihong et al., 2008, Hu et al., 2007, Hu et al.