Rats sacrificed by decapitation without anaesthesia had their bra

Rats sacrificed by decapitation without anaesthesia had their brain rapidly excised on a Petri dish placed on ice. The olfactory bulbs, pons, medulla, cerebellum, hippocampus and striatum were discarded, and the cerebral cortex was dissected, weighed and homogenized in 10 volumes (w/v) of 20 mM sodium phosphate buffer, pH 7.4, containing 140 mM KCl. Homogenates were centrifuged at 750g for 10 min at 4 °C to discard nuclei and cell debris ( Evelson et al., 2001). The pellet was discarded and the

supernatant, a suspension of mixed and preserved organelles, including mitochondria, was pre-incubated at 37 °C for 1 h with Prist at concentrations of 1, 10, 100 Ruxolitinib clinical trial or 200 μM. Controls did not contain this fatty acid in the incubation medium. Immediately after incubation, aliquots were taken to measure TBA-RS, carbonyl formation, sulfhydryl content and GSH levels. In some experiments, antioxidants were co-incubated with supernatants at the following final concentrations: 10 μM Trolox (TRO, soluble α-tocoferol), 1000 μM GSH, 1000 μM N-acetylcysteine (NAC),

combination of 20 mU/mL superoxide dismutase (SOD) plus 20 mU/mL catalase (CAT), 750 μM Nω-nitro-L-arginine methyl ester (L-NAME) and 1000 μM melatonin (MEL). The chosen concentrations of the antioxidants were those capable to efficiently scavenge free radicals ( Halliwell and Gutteridge, 2007). TBA-RS was determined according to the method of Esterbauer and Cheeseman (1990). Briefly, Uroporphyrinogen III synthase 300 µL of cold 10% trichloroacetic acid were added Metformin mouse to 150 µL of pre-incubated cerebral cortex supernatants and centrifuged at 3000g for 10 min. Three hundred microliters

of the pre-incubated supernatants were transferred to a pyrex tube and incubated with 300 μL of 0.67% TBA in 7.1% sodium sulphate on a boiling water bath for 25 min. The tubes containing the mixture were allowed to cool on running tap water for 5 min. The resulting pink-stained TBA-RS was determined in a spectrophotometer at 532 nm. A calibration curve was performed using 1,1,3,3-tetramethoxypropane, and each curve point was subjected to the same treatment as supernatants. TBA-RS values were calculated as nmol/mg protein and represented as percentage of control. Protein carbonyl formation, a marker of protein oxidative damage, was measured spectrophotometrically according to Reznick and Packer (1994). Two hundred microliters of the aliquots from the pre-treated supernatants were treated with 400 µL of 10 mM 2,4-dinitrophenylhidrazine (DNPH) dissolved in 2.5 N HCl or with 2.5 N HCl (blank) and left in the dark for 1 h. Samples were then precipitated with 600 μL 20% trichloroacetic acid and centrifuged for 5 min at 9000g. The pellet was then washed with 1 mL ethanol:ethyl acetate (1:1, V/V) and dissolved in 550 μL 6 M guanidine prepared in 2.5 N HCl at 37 ºC for 5 min. The difference between the DNPH-treated and HCl-treated samples (blank) was used to calculate the carbonyl content determined at 365 nm.

It is fairly obvious that the current CFP framework does little t

It is fairly obvious that the current CFP framework does little to fulfil these conditions. In addition to the problem of fragmented and uneven development of stakeholder organizations across Member States, the top-down style of micro-management is not ALK signaling pathway conducive to the development of industry partners ready to take on management responsibilities. Whereas the establishment of Regional Advisory Committees (RACs) and the involvement of national and/or transnational producer organizations (POs) in quota management are steps in the right direction, there seems to be a long way to go in developing strong industry partners

capable of taking on a comprehensive role as operators in a RBM system. Further, the responsibility for resource conservation, as set forth in the Treaties [67], leaves very little room for delegating management responsibility, be it to regional management bodies or industry partners. Further, there has been little movement in the direction of and cost recovery and of sharing or reversing the burden of evidence. Finally, while there are movements towards ITQ-like systems in some EU fisheries, strong arrangements for securing rights and privileges of resource users are absent in most cases.Resource

users, and their organizations may therefore lack sufficient motivation for investing in management and research through RBM like arrangements. As this suggests, the current CFP framework is not Ipilimumab cost conducive Montelukast Sodium to the development of RBM practices. To the extent that cases with RBM-like features can be found, these are at best partial, as in the cases of stakeholder initiation of management plans or in the implementation of recovery plans, or do not involve RBM in an organizational sense, as in the case of CQM. To which extent will the current CFP reform be able to change this state of affairs and construct a framework better suited for the RBM model? Given the thrust of the Green Paper, in particular its emphasis on RBM as

an approach that could repair the structural weaknesses of the CFP, this appears as a possibility. On the other hand, the CFP is strongly committed to ideas that are incongrous with RBM forms, and previous reform attempts have demonstrated that it does not change easily. Since it is not yet clear how the reformed CFP will be implemented in detail, definitive answers cannot be given to this question. The final compromise text on the Basic regulation on the CFP [68] and the Market Organization [69], however, include elements on RBM, although in a significantly changed form than the Commission envisaged in its Green Paper: The new CFP emphasizes the importance of developing multiannual management plans.

The wind effects are directly related to the pressure distributio

The wind effects are directly related to the pressure distribution over an area. However, as shown by tide gauge records, true sea level surges and falls can be several times higher than the values resulting from the action of tangential wind stress upon a fluid surface (Wiśniewski & Holec 1983). Suursaar et al. (2003) pointed out that the highest surge events on the west Estonian coast are associated with deep depressions producing strong SW and W winds in suitably oriented bays such as Pärnu Bay. An example is the mid-latitude

depression Gudrun, which occurred in January 2005 and caused the heaviest storm surge along the coasts of the Gulf of Riga (Suursaar et al. 2006). The sea level at Pärnu was 2.75 m higher selleck chemicals than the mean level there. In the Gulf of Finland, record increases in sea level were measured

as well, e.g. at Helsinki (1.51 m). Skriptunov & Gorelits (2001) NVP-LDE225 showed that significant wind-induced variations in the water level near the River Neva as well as their magnitude and duration result from the wind regime and the morphology of the near-mouth offshore zone. Averkiev and Klevanny, 2007 and Averkiev and Klevanny, 2010 analysed the effects of atmospheric pressure as well as wind direction and speed on the sea level in the Gulf of Finland. They showed the low pressure system trajectory to be potentially important in generating storm surges particularly damaging for St. Petersburg (Russia). The problem of sea surface deformation by concentric, mesoscale, fast-moving deep low-pressure systems was addressed by Lisowski, 1960, Lisowski, 1961 and Lisowski, 1963, Wiśniewski, 1996, Wiśniewski, 1997 and Wiśniewski, 2003, Wiśniewski & Holec (1983), Wiśniewski & Kowalewska-Kalkowska (2007) and Wiśniewski and Wolski, 2009a and Wiśniewski and Wolski, 2011. It seems, however, that this Rapamycin factor has been generally underestimated, even downright ignored, in the literature, a situation that has been detrimental to attempts at explaining mechanisms

of such extreme phenomena as coastal floods or low sea levels that adversely affect navigation safety, stability of hydraulic engineering structures, etc. It is true that a lowered atmospheric pressure system (a tropical cyclone or a concentric low pressure system) overlies a water cushion, moving together with the pressure system at the sea surface. Wave height depends on the pressure decrease in the centre of the system. A pressure drop of Δp = 1 hPa results in a static sea level rise of ΔHs = 1 cm under a stationary low ( Figure 1a, formula  (3)). When the depression moves over the sea surface, the latter becomes dynamically deformed (ΔHd). The sea level deformation shows positive wave elevations in the centre and negative elevations on the flanks of the deformation ( Figure 1b, formula  (4)).

Continuous variables were compared using Wilcoxon rank sum test (

Continuous variables were compared using Wilcoxon rank sum test (since non-normally distributed). Characteristics of the tests were analysed in 2 x 2 tables using the ‘diagt’ routine.20 The McNemar test was used to compare sensitivities of the tests. Two hundred and twenty nine patients were examined. All patients met the WHO clinical case definition for acute dengue infection.21 One hundred and sixty two patients (71%) returned for the follow-up specimen collection visit and were considered further. Of the 162 patients

included in the current analysis, 72 patients (44%) were given a laboratory confirmed diagnosis of dengue infection by paired serology (Table 1). Four patients were found to have evidence of recent dengue infection. The demographic and clinical data of the patients are shown in Table 2. Patients with confirmed dengue infection had lower white cell counts (4.8 vs 7.2, P<0.0001) ICG-001 chemical structure and platelets (147 vs. 162, P=0.03) than patients with non-dengue infection. Twenty six patients (16%) were found to have non-dengue causes for their illness: four patients (2%) grew Salmonella typhi from blood cultures, by serology nine patients (6%) had murine typhus, seven (4%) had scrub typhus, and six (4%) had leptospirosis.

Only one patient had evidence of dual infection: acute secondary dengue and typhoid; this patient had positive NS-1 antigen and dengue rRT-PCR and also grew Salmonella typhi from a blood KU-57788 cell line culture. The serotype of Casein kinase 1 dengue virus was sought by performing nested RT-PCR on the acute specimens from the 72 serologically

confirmed cases. Seventy one of the cases (99%) were serotype 3, with the remaining case serotype 2. The four patients with serological evidence of recent dengue infection were negative in the nested RT-PCR. The performance characteristics of NS-1 antigen detection, rRT-PCR, and IgM antibody detection in the acute plasma specimen against our ‘gold standard’ of paired serology are shown in Table 3. NS-1 antigen or IgM antibody test alone had low sensitivity compared with the paired serology result, 54% and 17% respectively. rRT-PCR sensitivity was 89%, significantly higher than both NS-1 antigen and IgM antibody tests (P<0.00001). Specificities of NS-1 antigen detection, rRT-PCR, and IgM antibody detection were 100%, 96% and 88% respectively. The effect of fever duration at presentation on assay sensitivity is shown in Figure 1. NS-1 antigen sensitivity peaked in the early stages of fever (three days of fever at presentation). IgM antibody sensitivity rose later, peaking in patients presenting with five days of fever. The sensitivity of rRT-PCR remained high throughout. However, as a result of the relatively small number of specimens on each day, particularly for four and five days of fever, the 95% confidence intervals around the sensitivities are wide. The performance characteristics of combinations of the acute specimen tests are shown in Table 3 (combined by an ‘OR’ operator).

Epidemiological studies are therefore needed on the distribution

Epidemiological studies are therefore needed on the distribution learn more and virulence potential of these yeasts in different population groups, addressing risk factors and developing strategies for the control and prevention of infections. 27, 30 and 31 Yeasts are found colonizing various sites in the oral cavity (lingual, palate, tonsils, mucosa of the lips and cheeks,

caries, periodontic and endodontic lesions). 32, 33, 34 and 35 Siqueira and Rôças 35 found C. albicans species associated with bacteria in teeth with periodontal pockets around areas of root exposure. For those authors, resistance to intra canal drugs, and the ability of these yeasts to colonize and invade the dentine tubules, may explain the presence of yeast in persistent endodontic infections. The use of a prosthesis is another factor that may favour colonization of the oral cavity by Candida spp. 36 with a report indicating that the microbiota between a prosthesis and palate mucosa has a composition similar to dental biofilm, except for a greater proportion of Candida species, a fact related to the development of candidiasis on the mucosa of the palate. Kleinegger et al.37 concluded that a number of natural barriers

existed in the mucosal surfaces and body fluids; preventing the colonization in healthy individuals. Protease Inhibitor Library concentration These barriers are more or less effective, depending on factors related to age, gender, smoking, diet, drugs and the host immune status. This explains the fact that not all individuals harbour Candida spp. Saliva helps maintain oral health, provides a buffering capacity and provides lubrication of the mucous membranes; therefore, qualitative and quantitative changes in saliva inevitably affect the physiology, defence mechanisms and microbial ecology of the mouth.38 Lactoferrin and lysozyme are two proteins in the innate immune response present in saliva and exert an antifungal modulating effect on the implantation of species of Candida in the oral cavity. 39 Other important proteins in human saliva that have a cytotoxic action on bacteria and fungi are the histatins, estaterins, lactoperoxidase

and calprotectin. 37 According to Lin et al., 40 when there is a decrease when the concentration of salivary histatins, dysfunction much of these proteins occurs and candidiasis tends to manifest. HIV-infected individuals show a reduction in salivary flow and an anti Candida activity of saliva and are often suffering from oropharyngeal candidiasis. For those authors, the saliva contained mucins and aggregated IgA, histatin, lactoferrin and lysozyme, which remained focused on mucosal surfaces and exerted an antimicrobial effect. 41C albicans is able to connect to several species of streptococci (S. oralis, S. sanguinis, S. gordonii, and Fusobacterium) through recognition receptor polysaccharides in the bacterial cell surface. F.

These findings agree with previous studies (Hasegawa et al , 1997

These findings agree with previous studies (Hasegawa et al., 1997 and Hasegawa et al., 2000) showing that CV treatment increased mRNA levels for granulocyte–macrophage colony-stimulating factor (GM-CSF). This stimulus can be attributed to the presence of a glycoprotein, which is purified from CV, is soluble in water and has been reported

to be a hematopoietic stimulator that increases CSF levels and promotes progenitor cell migration from the bone marrow to the spleen followed by an expansion GW-572016 order of CFU-GM in this organ after chemotherapy (Konishi et al., 1996). The presence of α-tocopherol in CV, the former of which is a member of the vitamin E family and possesses numerous biological properties including significant effects on inflammation, cell proliferation, and apoptosis (Azzi, 2007, Lemaire-Ewing et al., 2010 and Singh et al., 2006), may also be important here, as it has been shown to increase the number of HP as demonstrated by CFU-GM assays in the bone marrow of irradiated mice after treatment (Bichay and Roy, 1986, Cherdyntseva et al., 2005 and Roy et al., 1982). The presence of these components in CV can explain, in part, the fact that we observed a small but significant increase in CSA in the BM of non-stressed animals after CV treatment; however, this increase Selleck BTK inhibitor did not interfere with the number of HP or with the CFU-GM. The reduced capacity of cultured cells to support the growth and differentiation of CFU-GM following

the application of SST or RST was consistent throughout the duration of the cultures (7 weeks), and the suppression caused by SST was more severe until the 7th week. From the 1st to the 4th weeks of culture, the stromal layer is formed in the flasks. In the 5th week, the cultures are repopulated with cells from the respective groups of mice. These cells interact with the stroma, demonstrating their capability to maintain hematopoiesis. Therefore, we propose that SST and RST directly interfere with the physical contacts between stromal and hematopoietic cells. This hypothesis is in agreement with a significant reduction in the

local 3-oxoacyl-(acyl-carrier-protein) reductase production of IL-6 and IL-1α by stromal cells after stressor application, as observed in this study. IL-6 plays a critical role in the generation and maintenance of myelopoiesis in murine LTBMC (Hauser et al., 1997) and is a survival factor for hematopoietic stem cells (Bernard et al., 1994). Both IL-6 and IL-1α have synergistic activity with CSFs in stimulating hematopoiesis, thus contributing to the maintenance of neutrophil maturation and viability (Eaves et al., 1991, Dinarello, 1996 and Muench et al., 1992). Studies in the literature demonstrate that IL-1α accelerates both granulopoietic and thrombopoietic recovery in 5-fluorouracil myelosuppressed mice (Kovacs et al., 1997). However, in contrast to what we observed with HP and CFU-GM numbers, the decrease caused by SST and RST on the levels of these cytokines was of equal magnitude.

Moon et al (2012) concluded that despite the methodological shor

Moon et al. (2012) concluded that despite the methodological shortcomings, the evidence supports a causal relationship between high arsenic exposure and CVD, but remains inconclusive for low levels of exposure. Recent systematic reviews of hypertension likewise report that heterogeneity among studies limits conclusions regarding the consistency of the evidence. A meta-analysis of cross-sectional studies on arsenic exposure, hypertension, and blood pressure reported ZD1839 research buy a pooled

odds ratio comparing the highest with the lowest exposure groups in eight studies of 1.27 (95% CI: 1.09–1.47; p-heterogeneity = 0.001) ( Abhyankar et al., 2012). Paradoxically, the five studies with moderate to high exposure

yielded a non-significant pooled odds ratio with significant heterogeneity (OR = 1.15, 95% CI: 0.96–1.37; p-heterogeneity = 0.002), whereas the three low exposure studies (average <50 μg/L in drinking water) showed a clearer association with arsenic (pooled OR comparing the highest with the lowest exposure categories = 1.56, 95% CI: 1.21–2.01; p-heterogeneity = 0.27). The few studies that evaluated changes in systolic and diastolic blood pressure by arsenic exposure levels reported inconclusive findings ( Abhyankar et al., 2012). Similar findings of an elevated risk with considerable heterogeneity were reported in a second meta-analysis of arsenic exposure and hypertension ( Abir et al., 2012). An additional cross-sectional study from West Bengal, India, reported increased prevalence of hypertension

in a region with a mean well water arsenic concentration Depsipeptide of 50 μg/L (broad range of <3–326 μg/L) compared to a region with <3 μg/L (OR = 2.87; 95% CI: 1.26–4.83) ( Guha Mazumder et al., 2012). The Strong Heart prospective cohort study suggested that low arsenic exposure may be associated with CVD risk (Moon et al., 2013), although inconsistent results for iAs limit their use in dose–response assessment. Associations in never smokers but not smokers, and in those with greater amounts of DMA in urine, but not iAs and MMA, are in conflict with other Terminal deoxynucleotidyl transferase studies (e.g., Chen et al., 2001, Chen et al., 2011, Chen et al., 2013a, Tseng, 2009 and Wu et al., 2006) and with the mechanistic understanding of the toxicity of iAs and its metabolites ( Cohen et al., 2013). The urinary arsenic associations reflect ingestion of DMA or organic precursors (e.g., arsenosugars) in the diet rather than ingestion and metabolism of iAs. Moon et al. (2013) note that grains are a major source of dietary iAs; however, grains also supply DMA based on their low percentage of arsenic as iAs (11%, corn meal; 28%, wheat flour; 24%, rice; Schoof et al., 1999). Ingested DMA and organic precursors are considerably less toxic than iAs, particularly at low doses in the diet ( Cohen et al.

The ethanol yield from fungal pretreated rice straw in SSF alone

The ethanol yield from fungal pretreated rice straw in SSF alone was 67.1% (untreated RS, 23.4%; and EBI-RS, 61.4%) of the theoretical maximum yield of ethanol (Fig. 2). In addition, during the WEBI pretreatment, the loss of three main components (glucan, xylan, and lignin) and a total mass loss (w/w) in RS were negligible within the error range as they were <5% (i.e., <0.5 g) of the indices of evaluation (% glucose and % ethanol). In order to upgrade traditional Sirolimus datasheet EBI, RS was pretreated to improve the hydrolysis yields by using a water-based electron beam at 0.12 mA – 80 kGy

– 1 MeV. Based on the mass balance and the optimal WEBI (water soaking ratio of 100%) conditions, pretreated RS showed increases in the enzymatic hydrolysis (70.4% of the theoretical maximum) of cellulosic substrates as well as in ethanol production (67.1% of the theoretical maximum) in SSF, compared with those of the untreated RS. Structural composition analysis revealed that physical changes in lignocellulosic surfaces were most likely a result of WEBI. Quite importantly, the cost-effective

yields resulting from the WEBI pretreatment were not lower than those resulting from the physicochemical programs, and inhibitors were rarely generated. However, no “physicochemical programs” (i.e., Alectinib cost benchmark pretreatment runs) were included in the study. This work was supported by the by the Ministry of Education, Science and Technology, Republic of Korea. “
“Erythropoiesis is one of the body’s most productive cell proliferation processes, yielding an average of 2 × 1011 new erythrocytes from hematopoietic stem cells of the bone marrow every day to replace those lost to senescence and destruction [25]. A reduced erythropoietic output or the production of malfunctioning erythrocytes leads to anemia which

can have severe and even fatal consequences when tissues are insufficiently supplied with oxygen [17]. Homeostasis of erythrocyte production is primarily regulated by the hormone erythropoietin (EPO), whose production is upregulated upon tissue oxygen depletion [9] and [30]. However, numerous factors – both exogenous (such as toxins) and endogenous (such as inflammatory cytokines) – can inhibit proliferation and/or differentiation of erythroid cells [27]. In addition, the requirement for large amounts Miconazole of iron for hemoglobinization makes the process highly dependent on the availability of sufficient concentrations of transferrin-bound iron [16]. In diseases of chronic inflammation such as rheumatoid arthritis, erythropoiesis is impaired both by the direct action of proinflammatory cytokines such as tumor necrosis factor (TNF)-α and interferon (IFN)-γ and by upregulation of the liver hormone hepcidin, the primary regulator of iron uptake and storage, leading to a reduction in the amount of bio-available iron in circulation [10].

55 ml of acetone and 63 μl of concentrated HCl (37%) in capped Ep

The mixture was vortexed vigorously and then centrifuged at 24,462g for 10 min at 4 °C. The supernatant was extracted and the absorbance was measured at 407 nm against a reagent blank. Two replicates Nivolumab were measured, myoglobin solutions were used to make a linear standard curve and hemin concentrations were read from

the standard curve. Meat samples were placed into 16 × 125 mm screw-cap Pyrex culture tubes and 0.8 ml of the C13:0 internal standard, 0.56 ml of 10 N KOH in water, and 4.24 ml of MeOH were added. All tubes were incubated in a 55 °C water bath for 1.5 h with hand-shaking for 5 s every 20 min to properly permeate, dissolve and hydrolyse the samples. The samples were cooled to below room temperature and 0.464 ml of 24 N H2SO4 was added. All the tubes were incubated again in a 55 °C water bath for 1.5 h with hand-shaking for 5 s every 20 min; then the tubes were cooled again in a cold water bath and 2.4 ml of n-hexane were added to each tube. All the tubes were vortex-mixed for 5 min and centrifuged for 5 min in a table top centrifuge. The hexane layer, containing the fatty acid methyl esters, was transferred into a GC vial, capped and kept at −20 °C prior to GC analysis ( O’Fallon, Busboom, Nelson, & Gaskins, 2007). The fatty acid composition of the meat samples

was determined by gas chromatography on a fused capillary column. The oven temperature was 70 °C at the start, PR-171 solubility dmso held there for 4 min and then increased to 160 °C at a rate of 20 °C/min. Thereafter the temperature was held for a further 15 min, then the temperature was further increased at 3 °C per minute to 230 °C. Helium was used as the carrier gas at a flow rate of 68.4 ml/min at a temperature of 280 °C and the column head pressure was 309.4 kPa. Both the injector and

the detector were set at 260 °C. The split ratio was 30:1. The flame ionisation detector temperature was 290 °C with H2, air and N2 make-up gas flow rates of 40, 450 and 45 ml/min, respectively. The run time for a single sample was 92 min. C13:0 was added Hydroxychloroquine in vitro as an internal standard and used to calculate the amounts of fatty acids in muscle (mg/100 g). The fatty acids were identified by comparing their retention times with the fatty acid methyl standards. Minitab (version 16; Minitab Inc., State College PA, USA) was used for univariate regression analysis (incl. stepwise regression) and one way ANOVA. The unscrambler (version X 10.2 CAMO Software AS, Oslo, Norway) was used for principal component analysis (PCA), as well as partial least square (PLS) regression. Evaluation of the PLS regression model was with full cross-validation. Beef and chicken meat samples were incubated for different times, with or without liposomes, to examine when the largest amount of peroxides was formed. The peroxides in raw beef and chicken homogenates increased rapidly during the first 2 h of incubation at 37 °C.

The enzyme was completely inhibited by iron chloride, silver nitr

The enzyme was completely inhibited by iron chloride, silver nitrate and SDS in both concentrations tested. In the assay conditions, the denaturing action of SDS probably affected the integrity of the enzyme tridimensional structure which is fundamental for its catalytic activity. Inhibition caused by SDS (1 and 10 mM) was also demonstrated by Li, Jiang, Fan, and Liu (2012) for cloned β-glucosidase using metagenomic DNA from mangrove Torin 1 manufacturer soil. D. hansenii UFV-1 β-glucosidase activity was greatly increased by β-mercaptoethanol,

glucose, urea and aluminium chlorid at both concentrations tested. Non-inhibition by EDTA implies that divalent cations are not essential to enzyme activity ( Chen, Li, & Zong, 2012b) and it is not a metalloenzyme. Many works reported that EDTA does not inhibit β-glucosidases as in the case of Pyrococcus furiosus β-glucosidase that was considered metal-independent ( Yeom et al., 2012). β-Mercaptoethanol was the agent which best promoted enzyme activation in both final concentrations tested. The activation by this Tenofovir datasheet reducing agent can be explained

by the fact that some reduced chemical ligations in the enzyme structure are favourable for the catalytic activity. Calcium and magnesium have a stimulatory effect on D. hansenii UFV-1 β-glucosidase. It has been reported that these two ions are enhancers of β-glucosidase activity ( Oyekola, Ngesi, & Whiteley, 2007). Glucose was found to be a competitive inhibitor of D. hansenii UFV-1 β-glucosidase and the Ki value was 11.36 mM. In general, β-glucosidases are inhibited

by glucose and this inhibition is competitive ( Yang et al., 2004). Soy molasses is a by-product generated in the production of soy protein concentrate, in which isoflavones and other phytochemicals are enriched all (Hosny & Rosazza, 1999). This by-product in the soy industry is used as an inexpensive animal feed, but the processing and use of soy molasses as a functional food has been suggested (Najafpour & Shan, 2003). The potential of D. hansenii UFV-1 intracellular β-glucosidase to hydrolyze isoflavones in soy molasses to their aglycon forms was demonstrated for the free β-glucosidase and the alginate immobilised cells containing this enzyme ( Table 4). Prior to hydrolysis, glucoside isoflavones were predominant in the soy molasses, representing approximately 80%, where aglycones made up about 10%. After 2 h of treatment with the free or immobilised β-glucosidase the isoflavone glucosides were almost completely hydrolyzed after which there remained about 3% of these compounds. There was no change in the amounts of isoflavone glucosides and aglycones after 4 or 8 h of enzymatic treatment of soy molasses compared to the assay after 2 h of hydrolysis (data not shown). This indicates that a short incubation period is preferred over a prolonged incubation with the free or immobilised enzyme.