Pregnancy may affect drug Epacadostat metabolism including the induction of hepatic and gastrointestinal metabolic enzymes [2,3]. For example, cytochrome p450 (CYP) metabolism changes with mean increases of 35% reported for the activity of CYP3A4, the primary isozyme responsible for lopinavir (LPV) biotransformation [2]. Consistent with these changes, we previously reported a 28% decrease in LPV plasma exposure,

as estimated by the area under the plasma concentration vs. time curve (AUC) during third-trimester pregnancy (antepartum, AP) compared to post-partum (PP) in 17 HIV-1-infected pregnant women receiving a standard LPV/r dose of 400/100 mg twice daily (bid) [4]. More recently, we have confirmed that increasing the LPV dose during pregnancy to 533/133 mg bid offsets the reduced exposure we previously observed [5]. Pregnancy may also be associated with a decrease in protein binding (PB) of drugs in plasma due to dilutional decreases in albumin and α-1 acid glycoprotein (AAG) concentrations and the displacement of drugs from binding Tacrolimus research buy sites by steroid and placental hormones [6–8]. LPV is highly bound to plasma proteins including albumin and AAG with binding of >99%. Pregnancy potentially alters this binding to clinically relevant proportions such that small changes in PB associated with pregnancy may cause large changes

in the percentage of unbound drug (fraction unbound; FU). Unbound drug is the pharmacologically active component of total drug concentrations and the fraction of drug free to traverse membranes and exert therapeutic effect. An increase in LPV

FU during pregnancy may partially offset the decrease in total drug concentrations observed with standard dosing [4]. Our primary objectives were to (a) measure the PB of LPV during the third trimester of pregnancy (AP) and PP, (b) determine FU of LPV AP and compare to PP estimates, (c) assess whether AAG or albumin concentration correlate with FU and (d) assess whether LPV total drug concentrations influence FU. International Maternal Pediatric Adolescent AIDS Clinical Trials Group (IMPAACT) Protocol 1026s (P1026s) is an ongoing, prospective, nonrandomized, unblinded, multi-centre study of the pharmacokinetics of currently Teicoplanin prescribed ARVs used by HIV-1-infected pregnant women. P1026s is a sub-study of P1025, a prospective cohort study of HIV-1-infected pregnant women receiving care at IMPAACT sites. This report describes only the PB results for those women who were prescribed LPV/r 133/33 mg soft gel capsules (SGC). Results on the pharmacokinetics of total LPV for these women have been published separately [4,5]. Eligibility criteria for the LPV/r arm of P1026s were: enrolment in P1025, age ≥13 years, initiation of LPV/r as part of clinical care before 35 weeks’ gestation and intent to continue the current regimen until at least 6 weeks PP.

Plant-parasitic nematodes are one of the most important plant pathogens, causing extensive damage to a wide variety of economically important crops. The annual losses in agriculture resulting from this pest amounted to $125 billion worldwide in past years (Sasser & Freckman, 1987; Oka

et al., 2000). Chemical insecticides of nonselective nature possessing rapid nematicidal effects are widely used as control measures against these pathogens. However, the potential negative impact on the environment and ineffectiveness after prolonged use have led to banning or restricting of the use of most nematicides. Therefore, identification of safe and effective nematicides is urgently Selleckchem Anti-infection Compound Library needed and biocontrol measures have recently been given much attention as viable options (Schneider et al., 2003). Bacteria have shown great potential as biological interventions for controlling nematode infections (Tian et al., 2007). Bacteria can affect nematodes via two primary mechanisms of action: direct obligate parasitism and indirect effects. Nematode parasitism is characteristic of Pasteuria spp.,

which are unusual mycelial, obligate, endospore-forming bacteria that can penetrate selective HDAC inhibitors the bodies of nematodes (Dong & Zhang, 2006; Tian et al., 2007). Some Pasteuria spp. have been used as a nematode control strategy (Sayre & Starr, 1985; Sayre et al., 1988, 1991; Giblin-Davis et al., 2003; Bishop et al., 2007). Different bacterial species (e.g. rhizobacteria) have antagonistic properties that affect nematode viability, including toxin production, metabolic-by-products that affect nematode viability, the production of damaging enzymes and nutrient competition (Siddiqui & Mahmood, 1999; Dong & Zhang, 2006; Tian

et al., 2007). The Pseudomonas fluorescens strain CHA0 extracellular protease AprA has been shown to possess biological activities against Meloidogyne incognita (Siddiqui et al., 2005). The Brevibacillus laterosporus G4 and the Bacillus nematocida protease demonstrated nematicidal effects when used on Bursaphelenchus xylophilus (Huang et al., 2005; Niu et al., 2006; Tian et al., 2006). Toxins suppressing nematode function have also been reported (Jacq & Fortuner, 1979; Ali et al., 2002; Jagdale & Grewal, 2002). In addition, Bacillus firmus metabolites FER generated during fermentation resulted in the death of parasitic plant nematodes (Mendoza et al., 2008). Furthermore, metabolites including 2,4-diacetylphloroglucinol (2,4-DAPG) were shown to control cyst and root-knot nematodes (Cronin et al., 1997; Siddiqui & Shaukat, 2003). Gram-positive bacteria belonging to the genus Bacillus are aerobic, endospore-forming organisms belonging to the plant growth-promoting rhizobacteria. Numerous reports have suggested that some Bacillus strains possess nematicidal properties (Kloepper et al., 1992; Krebs et al., 1998; Siddiqui & Mahmood, 1999; Siddiqui, 2002; Li et al.

Phylogenetic analysis of the gene sequences was determined using the maximum parsimony program included in paup* 4.063a (Swofford,

1998). Sequences were visually aligned for analysis and Saccharomyces cerevisiae was the designated outgroup species. In the present study, 26 strains representing 19 species of the Starmerella clade were analyzed for production of sophorolipids. Results are reported in Fig. 1, which gives the yield for strains of each species, and the phylogenetic placement of the strains as determined from the analysis of D1/D2 LSU rRNA gene sequences. Five of the 19 species tested showed significant production of sophorolipids: C. apicola, S. bombicola, learn more Candida riodocensis, Candida stellata and a new species of Candida, NRRL Y-27208, which will be described in a future study. In our Selleck NVP-BGJ398 earlier work, phylogenetic analysis detected 12 species in the Starmerella clade (Kurtzman &

Robnett, 1998) and they separated into two subclades, one represented by C. bombicola and the other by C. magnoliae. With the widespread application of gene sequence analysis in yeast taxonomy, 41 separate lineages (species) are known for the clade and all were included in the phylogenetic analysis shown in Fig. 1 to lend perspective to the placement of species that were tested for the biosynthesis of sophorolipids. However, many of the lineages are undescribed species, which are recognized only from their gene sequences, and cultures are not presently available for analysis. Even with the addition of many
ages to the Starmerella clade, the two subclades originally recognized are still evident. Based on the present analysis, sophorolipids are produced only by members of the S. bombicola subclade. Although not included in our analysis, C. batistae was shown by Konoshi et al. (2008) to form sophorolipids, and this species is a member of the S. bombicola subclade (Fig. Montelukast Sodium 1). As seen from Fig. 1, not all members of the subclade produce sophorolipids, and of particular interest for C. apicola, NRRL Y-2481 gave the greatest yield of any strain tested, whereas

NRRL Y-6688, a somewhat divergent strain of this species, produced essentially no sophorolipids. In earlier studies of sophorolipid biosynthesis by C. apicola, Tulloch et al. (1968) reported a yield of 40 g L−1 without optimizing the culture medium, much as we found in our assays. Our goal in this study was to test previously unexamined species for sophorolipid production without optimization. We did, however, examine the effect of incubation time, shaker speed and glucose concentration on sophorolipid production by C. bombicola NRRL Y-17069 and Candida sp. NRRL Y-27208, which as described below, produce sophorolipids with a different molecular structure. Starmerella bombicola NRRL Y-17069 gave maximum sophorolipid yield after 144-h incubation, whereas Candida sp.

05, P < 0.0001; Fig. 7A). Tukey post hoc analysis revealed that the severe lesion was significantly different from the intermediate lesion (P < 0.05) and highly significantly different from the mild lesion (P < 0.0001), while the intermediate lesion was significantly different from the mild lesion (P < 0.05). Apomorphine-induced rotation was also able to differentiate between the three subgroups (Group, F2,33 = 15.09, P < 0.0001; Fig. 7B). The post hoc analysis revealed that the severe lesion was significantly different from the intermediate lesion (P < 0.05) and

highly significantly different from the mild lesion (P < 0.0001), while the intermediate lesion was significanly different from the mild lesion (P < 0.05). Amphetamine

rotation was clearly less informative and could only differentiate between the animals with a mild lesion and those with > 60% striatal denervation (Group, F2,33 = 10.69, http://www.selleckchem.com/products/INCB18424.html P < 0.0001; Fig. 7C); Tukey post hoc analysis revealed that the mild lesion was significantly different Selleck Doramapimod from both the intermediate and the severe lesions (P < 0.001 and P < 0.05, respectively). By contrast, neither the stepping test nor the cylinder tests were able to distinguish between any of the lesion types (Group, F2,33 = 2.08, P = 0.15, n.s; Group, F2,27 = 1.31, P = 0.29, n.s, respectively; Fig. 5D and E). A subset of seven severely lesioned mice was followed long-term in four of the tests that showed profound deficits at the early post-lesion time-point (6–7 weeks), and were compared to a group of seven intact control animals (Fig. 8A–D). In all four tests the two groups showed stable

performance over the entire test period (20–23 weeks), and the lesioned and intact mice performed significantly different from one another in all four tests, including the corridor test (Group, χ21,48 = 827.14, P < 0.0001; Fig. 8A), apomorphine-induced rotation (Group, χ21,48 = 159.69, P < 0.0001; Fig. 8B), amphetamine-induced rotation (Group, χ21,48 = 26.91, P < 0.0001; Fig. 8C) and the stepping test (Group, χ21,36 = 208.26, P < 0.0001; Fig. 8D). There was no significant effect Benzatropine of time measured in any of the behavioural tests, thus confirming the stability performance in both the intact and lesioned groups (data not shown). The results show that intranigral 6-OHDA lesions can be used to induce profound loss of midbrain dopaminergic (DAergic) neurons, accompanied by extensive denervation of the striatum and behavioural impairments in a range of drug-induced and spontaneous motor tests. Based on the extent of striatal TH+ denervation we allocated the mice into three subgroups, exhibiting severe, intermediate and mild lesions of the mesostriatal pathway. From the behavioural impairments seen in these subgroups, it was possible to predict the severity of the lesion, i.e.

The rate of treatment modification after failure was 51.6 per 100 person-years (95% CI 45.6–58.4). Of the 194 patients whose treatment was not modified after treatment failure, three patients died, and 18 patients were lost to follow-up. Time to treatment modification after failure, by country income category and by type of treatment failure, is shown in Figure 1. The rate of treatment modification was similar in patients from high- and low-income countries. However, the rate of modification was higher in patients with a virological failure than in patients with either immunological failure or clinical progression. At the end of the first year following

failure, approximately 40% of patients with virological failure remained on the previous Natural Product Library cost regimen, compared with over 60% of patients with either immunological failure or clinical progression. Table 2 shows the factors associated with time to antiretroviral treatment modification after treatment failure by univariate and multivariate Cox proportional hazards models. In the final model (stratified by TAHOD sites) the factors independently associated with treatment modification after failure included CDC classification, CT99021 CD4 cell count and HIV viral load, all at the time of treatment failure: compared with patients who were in CDC category A, patients in category C were more likely to have a modification of treatment [hazard ratio (HR) 1.38, CI

1.01–1.87, P=0.040]; compared with patients with a CD4 count ≤50 cells/μL at the time of failure, patients with a CD4 count ≥51 cells/μL were less likely have their treatment modified (HR 0.61, CI 0.40–0.93, P=0.022); lastly, compared with patients with an HIV load <400 copies/mL at the time of failure, patients with an HIV viral load ≥400 copies/mL or those with an unavailable HIV load were more likely to have their treatment modified (HR 2.69, CI 1.90–3.81, P<0.001; HR 1.74, CI 1.14–2.66, P=0.010, respectively). Overall, there was little difference between high-

and low-income sites in terms of time this website to treatment modification after failure. However, from Figure 1 it appears that there may be some divergence after 2 years. We therefore performed an additional stratified analysis, comparing high- and low-income countries in the first 2 years, and more than 2 years, after failure. In the first 2 years, the rate of modification was similar in low- and high-income countries (low- vs. high-income, HR 1.08, 95% CI 0.80–1.46, P=0.632). In follow-up after 2 years, the rate was lower in patients from low-income countries; however, possibly because of the small numbers of patients with up to 2 years of follow-up (90 in total), the difference was not statistically significant (low vs. high, HR 0.49, 95% CI 0.23–1.03, P=0.059). Sensitivity analyses were also performed on patients who started treatment in or after 2003; the results were similar to those obtained when all eligible patients were included (data not shown).

Dnmt2 was one of the first cytosine-5 RNA methylases identified in a multicellular organism [16••]. Although Dnmt2-mediated methylation of cytosine 38 in the

anticodon loop of tRNAAsp was conserved in plant, flies and mice, none of these organisms lacking the functional Dnmt2 protein displayed any morphological differences to their wild-type counterparts [16••]. In contrast, morpholino-mediated loss of Dnmt2 in zebrafish reduced the size of the morphants by half and specifically affected liver, retina and brain development due to a failure to conduct late differentiation [17]. Over-expression of Dnmt2 on the other hand prolonged the life span of Drosophila by more than 50% and increased the resistance to stress

[ 18]. In line with these studies, Drosophila Dnmt2 loss-of-function click here Etoposide mouse mutants showed reduced viability under stress conditions, and Dnmt2-mediated methylation protected tRNAs from stress-induced ribonuclease cleavage ( Figure 1a) [ 9]. Cleavage of tRNAs is a conserved response to several stress stimuli in eukaryotes and the tRNA fragments are produced to repress translation by displacing translation initiation and elongation factors from mRNAs or by interfering with efficient transpeptidation [19, 20 and 21]. However, whether and how increased tRNA cleavage in Drosophila Dnmt2 mutants is directly linked to stress tolerance and protein translation is currently unknown. While tRNA cleavage is mediated by angiogenin in mammals, the only identified tRNA nuclease in Drosophila so far is Dicer [ 22]. Interestingly, also Amrubicin expression of Dicer is down-regulated by oxidative stress and Dicer knockout cells can be hypersensitive towards oxidative stress whereas its over-expression confers stress resistance

[ 23]. Other functions that have been linked to Dnmt2 but may be independent of its tRNA methyltransferase activity are silencing of retro-transposons and control of RNA viruses in Drosophila as well as RNA-mediated paramutations in the mouse [ 24]. Together, these data implicate that Dnmt2 is functionally redundant for normal development of most multicellular organisms but implicated in cellular stress responses at least in adult flies [ 24]. At least two more enzymes NSun2 and NSun4 can generate 5-methylcytidine in RNA in mammals (Figure 1a and b) [25 and 26]. Both belong to the S-Adenosylmethionine (AdoMet)-dependent methyltransferase superfamily and at least five more putative m5C RNA methylases in mammals (NOP2, NSun3, and NSun5–7) are predicted to methylate RNA based on sequence conservation of key catalytic residues [12]. Although the substrate specificities are unknown, NSun1 and NSun5, in addition to NSun2 and Nsun4, have been identified as mRNA-binding proteins [27].

A coupled wave, astronomical tide and storm-surge model has been developed and applied to the Mediterranean Sea on unstructured grid. The third-generation WWMII spectral wave model has been coupled with the 3-D hydrodynamic SHYFEM model. The method used here, and the numerical schemes employed in both models have been successfully tested and showed to be efficient in simulating tides, storm surges and waves along the Italian 5-FU datasheet peninsula. This marine model uses, as atmospheric data input, forecast fields produced by a meteorological model chain, from global to local scale. The variable resolution of the method and the effect of the depth-varying loading factor lead the present model, at least

for the Italian coast and for period of test, to perform better than other tidal models. Tide-surge non-linear interaction turns out to improve significantly the tidal model performance. Moreover, it has been found that the use of a three-dimensional formulation enhances the results of the tide-surge model. Hindcast results showed that the hydrodynamic-wave model coupling slightly enhanced

the wave prediction, while wave effect on the water level could not be resolved properly since the resolution of the numerical mesh of this application is not enough to describe the surf zone along see more the whole Italian coast. The modelling system described in this work, which includes meteorological and oceanographic components, represents a powerful short term water level forecasting system for the Italian region. The high spatial resolution of

the Kassandra system along the Italian peninsula, exploiting unprecedented high resolution meteorological model input, allows the detailed description of the sea water level and the wave field. The developed model gives a significant improvement in predicting the total water level along the Italian coastal area and represents a potentially useful tool in bathymetry and altimetry corrections. Even if the forecast skill for the surge signal depends strongly on the range of the forecast, the total water level is PIK3C2G less depended on it. The short term storm surge forecasts of the Kassandra system for the whole Mediterranean are available at http://www.ismar.cnr.it/kassandra. The operational model has been recently implemented also in the Black Sea. The implementation of the baroclinic version of the model and the investigation of different surface wind stress parameterizations will be the subject of future work. The authors thank the Italian Institute for Environmental Protection and Research (ISPRA) for providing water level and wave data. Finally, the authors would like to thank Dr. Luigi Cavaleri for the critical review of the manuscript. This research was partially funded by RITMARE Flagship Project, funded by MIUR under the NRP 2011-2013, approved by the CIPE Resolution 2/2011 of 23.03.2011.

, 1998). Lactobacillus

paracasei subsp. paracasei NTU 101 was isolated from human feces, and it is resistant to gastric juice and bile salt in the natural environment ( Lin et al., 2004). It also has probiotic characteristics that are effective in reducing cholesterol in blood and in the liver ( Chiu et al., 2006). L. paracasei subsp. paracasei NTU 101 can upregulate the antigen-presenting ability of dendritic cells and expression of natural killer group 2 D molecules capable of triggering natural killer cell-mediated cytotoxicity in BALB/c mice ( Tsai et al., 2008). Hydrolysates of L. paracasei subsp. paracasei NTU 101 can induce proliferation of macrophages and splenocytes and a release of the cytokines IL-10 and IL-12 that modulate the innate and adaptive immunity Ceritinib nmr and an inflammatory response ( Chiang et al., 2012a). Soy skim milk fermented by L. paracasei subsp. paracasei NTU 101 (NTU 101F milk), with or without

a Momordica charantia supplement, is effective at preventing and retarding hyperlipidemia-induced oxidative stress and atherosclerosis in hyperlipidemic hamsters ( Tsai et al., 2009). This type of milk is also useful for prevention of acute gastric ulcers induced by pylorus ligation and acidified ethanol (via prostaglandin E2), and it significantly enhances superoxide dismutase (SOD) activity ( Liu et al., 2009). Moreover, NTU 101F milk can attenuate bone loss in ovariectomized (OVX) mice ( Chiang and Pan, 2011) and aging-induced bone loss in BALB/c mice and can Liothyronine Sodium lower the risk of osteoporosis ( Chiang et al., 2012b). In addition, NTU 101F milk can upregulate and www.selleckchem.com/products/pd-166866.html downregulate lipolysis and

heparin-releasable lipoprotein lipase, respectively, in 3T3-L1 cells and can reduce obesity in Wistar rats fed a high-fat diet ( Lee et al., 2013). Taken together, the above data show that intake of cultured probiotic L. paracasei subsp. paracasei NTU 101 is likely to have various beneficial effects on the health of humans and animals. Hence, the safety of L. paracasei subsp. paracasei NTU 101 must be evaluated. Accordingly, the aim of this study was to assess the possible genotoxicity and mutagenicity of Vigiis 101 powder made from L. paracasei subsp. paracasei NTU 101. In addition, we performed a 28-day oral toxicity assay in Wistar rats. Sodium azide, benzo[α]pyrene, mitomycin C, 2-aminofluorene, 2-amino-anthracene, 4-nitro-o-phenylenediamine, 9-aminoacridine, acridine orange, and cyclophosphamide monohydrate were purchased from Sigma (St. Louis, MO, USA). The main media, including McCoy’s 5A medium with 10% fetal bovine serum and penicillin-streptomycin solution, minimal glucose agar plates, master plates, soft agar, and nutrient broth, were obtained from Biological Industries (Kibbutz Beit-HaEmek, North District, Israel). The probiotic used in this study was Vigiis 101 (SunWay Biotecn Co., Ltd., Taipei, Taiwan). Vigiis 101 is a dry powder which contains 6 billion bacteria of L.

GMP consist of high molecular weight glutenin subunits (HMW-GS) linked with low molecular weight glutenin subunits (LMW-GS) through disulfide bonds [6]. HMW-GS play an important role in determining the glutenin protein network structure [5],

and LMW-GS may also have a specific effect on glutenin aggregation [4]. GMP consisting of a higher ratio of HMW-GS to LMW-GS is correlated with improved wheat flour quality [7]. Therefore, subunit composition and GMP characteristics determine the rheological properties of wheat dough, and a close correlation between GMP http://www.selleckchem.com/products/lgk-974.html characteristics and end-use quality has been shown. HMW-GS are encoded by polymorphic genes at the Glu-1 loci on the long arms of group 1 chromosomes. Hexaploid wheat usually contains 3–5 subunits, zero or one encoded by Glu-A1, one or two by Glu-B1 and two by Glu-D1 [8]. The content and size distribution of GMP in wheat grains are both genetically and environmentally controlled. Drought promotes HMW-GS accumulation

in the early grain filling stage, whereas the opposite effect occurs at late grain filling [9]. Increasing N fertilizer increases the proportion of GMP in wheat flour [10]. Clay soil results in the accumulation of HMW-GS and GMP when compared to loam soil and sandy soil [11]. When under high temperature stress during the kernel filling period, the contents of particular Glu-D1 HMW-GS in weak-gluten wheat are much more sensitive than that in strong-gluten wheat [12]. In recent years, frequent soil water stress in northern China has influenced both dry matter production Caspase activation and quality of wheat [13]. Increased N levels promoted the accumulation of HMW and LMW-GS, GMP content and proportion of larger GMP particles under irrigated conditions. Under rainfed conditions, increased N fertilizer also increased protein content [14]. Both dough development time and dough stability time were longest with a single post-anthesis irrigation, whereas a second irrigation led to shortened dough development and dough stability times and weakened gluten strength, as well as a decreased glutenin polymerization

index and average sized GMP [15]. However, information about the impact of different irrigation patterns Glutathione peroxidase on accumulations of GMP in wheat grain is still limited. Although numerous studies have been conducted on size distribution and properties of GMP particles in wheat grains, there is limited information about the size distribution of different quality types of wheat under irrigated and rainfed conditions. The objective of the present study was to investigate differences that may occur in GMP accumulation in field-grown wheat cultivars under irrigated and rainfed regimes. HMW-GS and GMP contents and GMP particle distributions in four wheat cultivars were therefore investigated. The experiment was conducted on the experimental farm of the Research Institute of Agricultural Science (37°N, 116°E), Dezhou, China.

The phytosterol mixture contained 46 g/100 g β-sitosterol, 26 g/100 g campesterol,

17 g/100 g stigmasterol and 11 g/100 g of others minor PS. Cocoa powder, butter and liquor (Barry Callebaut®, São Paulo, Brazil), palm oil (Agropalma®, selleck inhibitor Jundiaí, São Paulo), hazelnut paste (La Morela Nuts®, Tarragona, Spain), rice protein (Acerchem International®, Shangai, China), polydextrose (Winway®, São Paulo, Brazil), erythritol (Cargill®, São Paulo, Brazil), maltitol (Huakong®, São Paulo, Brazil), sucralose (Tate Lyle®, São Paulo, Brazil), nut aroma (IFF®, Taubaté, Brazil) and soy lecithin were purchased in a specialized market (São Paulo, Brazil). The antioxidants (ascorbic acid and α-tocopherol) were obtained from Sigma–Aldrich (St. Louis, MO, USA).

A chocolate formulation containing 50 g/100 g of cocoa was used to coat the filling and was provided by Chocolife Indústria e Comércio de Alimentos Funcionais Ltda (São Paulo, Brazil). Bis(trimethylsilyl)-trifluoracetamide (BSTFA) containing 1 g/100 g trimethylchlorosilane (TMCS), pyridine, cholesterol, 5β-cholestan-3α-ol (epicoprostanol), (24S)-ethylcholest-5,22-dien-3β-ol (stigmasterol), (24R) –ethylcholest-5-en-3β-ol (β-sitosterol), 24α-ethyl-5α-cholestan-3β-ol(stigmastanol),(24S)-methylcholest-5,22-dien-3β-ol SCH772984 in vivo (brassicasterol) and (24R)-methylcholest-5-en-3β-ol (campesterol) were purchased from Sigma–Aldrich (St. Louis, MO, USA). Control chocolates (CONT) were formulated mixing cocoa powder, cocoa liquor, palm oil, polydextrose, rice protein, cocoa butter, xylitol, maltitol, hazelnut paste, erythritol, soy lecithin, polyglycerol polyricinoleate, nut flavor, sucralose and nut flavor. In pheromone the PHYT and PHAN formulations, palm oil used to prepare the filling was replaced by PS esters. In the PHAN chocolates, ascorbic acid and α-tocopherol were also added into the filling formulation (0.90 mg/100 g of chocolate). Belgian pralines were produced in an industry pilot plant as one batch. Firstly, all fats were weighted and placed in the mixer to melt at 45 °C. Afterward, dried ingredients were added to the melted fats and the mixture was conched by

a runner mill at 60 °C/6 h, promoting the evaporation of undesirable flavors and water. The mixture was refined at 40–55 °C until an average particle size of 23 μm had been achieved. All samples were manually tempered in a cold marble surface until the temperature reached 29 °C. The chocolate was molded in plastic moulds (14 cm length and 13 mm height) to receive the filling. A thin layer of chocolate was placed in the mould, left to cool and added of 15 g of filling. PS and antioxidants were included in the filling to avoid the negative temperature effect on lipid oxidation during the coaching and tempering process. After cooling the filling at room temperature, another thin layer of chocolate was added to cover the filled chocolate. Thus, each bar (30 g) was composed of 15 g of shell and 15 g of filling.