5) In UA159, cystine starvation resulted in

5). In UA159, cystine starvation resulted in Selleckchem AZD6738 a lower growth yield as well as a longer doubling time (Tdc. 93.3 ± 0.7 min) compared with its growth in the presence of cystine (Tdc. 76.3 ± 1.5 min), indicating that l-cystine is required for optimal growth of S. mutans. However, growth was completely abolished in SmTycABC under cystine starvation. Supplementing the modified growth medium with 0.1 mM cystine slightly improved the drastic growth impairment of the SmTcyABC mutant (Tdc. 118.2 ± 0.8 min). Similar to the SmTcyABC transporter mutant, the TcyR-deficient mutant (SmTcyR) had a longer doubling time (Tdc. 117.2 ± 3.8 min)

under cystine-supplemented (1 mM) conditions relative to wild type (Fig. 5). In contrast to SmTcyABC, SmTcyR was able to survive under cystine-deficient conditions, although its doubling time was remarkably increased relative to wild type (Tdc. 261.0 ± 11.9 min). Also importantly, growth kinetics of SmTcyR revealed a notable increase in the lag time regardless of the presence or absence of cystine, compared with the wild-type UA159 and SmTcyABC. We further evaluated the effect on growth by individual components of the TcyABC operon by conducting growth studies on mutants deficient in each gene. Briefly, growth kinetics were monitored for the TcyA, this website TcyB, and TcyC

transporter mutants in modified MM without cystine (Fig. 6). The most drastic effect on growth was observed for SmTcyB. Similar to TcyABC, growth of this mutant was completely abolished without cystine. Although TcyA and TcyC were able to grow in cystine-deficient medium, their

growth was tremendously impaired relative to wild type as judged by their longer doubling times; Tdc. 131.3 ± 4.8 and 214.8 ± 21.5 min, respectively. Sperandio et al. 2010 also showed impaired growth in the form of pinpoint colonies when their TcyA mutant was grown in chemically defined medium with the addition of cystine as the sole sulfur source. However, they did not investigate the growth of other Tyc ABC mutants. The ability of some of our TycABC mutants to grow in the absence of cystine, albeit in an impaired fashion, suggests that the presence of other amino acids (i.e. glutamate and leucine), inorganic sulfur, and/or ammonium sources were sufficient to sustain growth. S. mutans possesses amino acid biosynthetic pathways and even though most amino acids are not freely available in the Acyl CoA dehydrogenase environment, some strains are able to synthesize all the necessary amino acids required for survival (Liu & Ferro-Luzzi Ames, 1998; Albanesi et al., 2005). The ability of S. mutans to scavenge and compete for limited nutrients in the plaque biofilm is an important aspect that confers an ecological advantage, which facilitates its survival and persistence in the oral cavity. The amino acid transport system in S. mutans UA159, encoded by the tcyABC operon that is induced under cystine-starved conditions, functions to maintain growth by transporting cystine into the cell.

It is possible that most of these patients get infected by contac

It is possible that most of these patients get infected by contact with a taenia carrier.

The time ACP-196 manufacturer elapsed between disease acquisition and symptoms occurrence suggests that, at least in some patients, clinical manifestations are related to reactivation of an infection that has previously been controlled by the host immune system. Neurocysticercosis is the most common helminthic infection of the nervous system, and a major cause of acquired epilepsy worldwide.1 The disease occurs when humans become intermediate hosts of the tapeworm Taenia solium by ingesting its eggs from contaminated food or, most often, directly from a taenia carrier (fecal-oral route). Within the central nervous system, parasites may lodge in the brain parenchyma, subarachnoid space, ventricular system, or spinal cord, causing a myriad of pathological changes that are responsible for the clinical pleomorphism of neurocysticercosis. Neurocysticercosis is endemic in most learn more of the developing world. There, millions of people are infected by cysticerci, and many of them will eventually experience the clinical consequences of this infection.2 Neurocysticercosis was rare in developed countries up to the past few decades. Together with the growing number of immigrants from endemic areas, there has been

an increase in the number of patients with cysticercosis in some of these countries.3,4 Also, increased tourism and international business affairs have rendered people from nonendemic areas more susceptible to acquire this parasitic disease. Neurocysticercosis in travelers has not been well characterized, and available information on these individuals is scarce and incomplete.5 The main purpose of this study is to present a review of the literature on neurocysticercosis in citizens from nonendemic countries who developed the disease after a travel to disease-endemic regions, to estimate the magnitude of the disease, and to describe the pattern of disease expression in this population. A literature search of neurocysticercosis occurring

Paclitaxel in citizens from nonendemic countries who had history of travel to disease-endemic countries over the past 30 years (1981–2011) was performed using the electronic database of MEDLINE (National Library of Medicine, Bethesda, MD, USA). Key words “cysticercosis” and “neurocysticercosis” were combined with “travel,”“traveler,” and with the name of each of the countries traditionally considered as nonendemic, including Western European countries, African and Middle-East countries of the Arab World, Israel, some American countries (Argentina, Belize, Canada, Surinam, United States, and Uruguay), Islands of the Caribbean Basin (except Haiti and Dominican Republic), and some countries of Asia and Oceania (Australia, Japan, Malaysia, and New Zealand).


P = 00015, and treatment effect: F2,20 = 1480, P =


P = 0.0015, and treatment effect: F2,20 = 14.80, P = 0.0002; n = 12 WT and 11 KO; Fig. 4A Adriamycin concentration and B]. Specifically, the level of phosphorylation increased in WT no extinction and extinction groups relative to the WT CS-only group (P < 0.05 and P < 0.01, respectively). The increase for the extinction group was also greater than for the no extinction group (P < 0.05). This was in contrast to the situation for PN-1 KO mice. As in the case for the WT, the no extinction group showed a significant increase in phosphorylation level over the PN-1 KO CS-only mice (P < 0.01); however, the extinction group did not. The WT extinction group pαCamKII/αCamKII ratios were also significantly greater than for the PN-1 KO extinction group (P < 0.01). These results suggest that the mITC cells are responsive to both fear retrieval and extinction acquisition. Similarly, the decreased GSK2126458 cost response in the mITC of PN-1 KO mice correlates with their impaired extinction behavior. The analysis of pαCamKII/αCamKII ratios in the lITC (Fig. 4C and D) showed no behavior-dependent changes in either WT or PN-1 KO mice. The overall levels for PN-1 KO groups, however, tended to be lower than for the corresponding WT group (genotype

effect: F1,21 = 6.760, P = 0.0187; n = 12 WT and 11 KO). We also examined pαCamKII/αCamKII ratios in two subdivisions of the CEA (Fig. 5). In the CEl, the WT and PN-1 KO extinction groups showed a significant increase in phosphorylation cAMP levels over their respective CS-only controls (genotype effect: F1,21 = 12.01, P = 0.0030, and treatment effect: F2,20 = 11.52, P = 0.0007; n = 12 WT and 11 KO; extinction compared with CS-only group: WT, P < 0.05 and KO, P < 0.01; Fig. 5A and B). The increase shown

by the PN-1 KO mice in the extinction group was significantly greater than the corresponding values for the WT extinction group (P < 0.05). While there were no significant changes in the no extinction groups compared with CS controls, there was an overall trend to increased phosphorylation levels in PN-1 KO compared with the WT mice. In comparison, analysis of pαCamKII/αCamKII ratios in the CEm (Fig. 5C and D), and in the LA and BA (supporting Fig. S3) showed that neither WT nor PN-1 KO values varied with the behavioral groups. Taken together, our data indicate that extinction triggers the phosphorylation of αCamKII specifically in the mITC and CEl, and that this response is perturbed in the PN-1 KO mouse. Our behavioral results indicate that fear extinction is severely impaired in PN-1 KO mice. This deficit is accompanied by an abnormal pattern of activity-dependent signaling markers across different amygdala nuclei, including the BA, mITC and CEl.

2e) Taken together, these results suggested that one possible me

2e). Taken together, these results suggested that one possible mechanism of Trichokonin-induced resistance against TMV is the induction of early plant defense reactions. To find out the mechanism involved in Trichokonin-induced resistance against TMV in tobacco, the activities of PAL, POD and SP600125 PPO were analyzed. These PR enzymes play key roles in tobacco resistance against TMV (Chen et al., 2009). As shown in Fig. 3a and b, the activities

of PAL and POD increased after Trichokonin treatment. On the fourth day of treatment, both PAL and POD reached their maximum activity, with the peak values of 8.4-fold (PAL) and 5.2-fold (POD) higher than in the control plants, respectively. After a 4-day treatment, the activities of these two enzymes began to decrease and showed a drastic decrease after a 5-day treatment. PPO activity showed a slight increase during a 6-day treatment with Trichokonins (Fig. 3c). Apparently, Trichokonin treatment could differentially influence the activities of PR enzymes. To gain further insight into the mechanism involved in Trichokonin-induced resistance against TMV, the transcription levels of selected plant defense genes were analyzed. As shown in Fig. 4, seven genes involved

Seliciclib in plant defense response were studied. A gene expression level that upregulated >1.5-fold (P<0.05) was considered a significant difference between control and Trichokonin treatment. SOD, CAT, APX and POX are known to be associated with the reactive oxygen intermediate (ROI)-mediated signaling pathway (Baker et al., 1997). Trichokonin treatment led to about 1.8-fold upregulation of SOD and CAT, 2.5-fold

of APX and 2.3-fold of POX genes, compared with the controls (Fig. 4a). Trichokonin treatment also upregulated the expressions of NtPR1a, a marker gene of the SA-mediated defense pathway (1.9-fold) (Fig. 4b). The expression of NtPR3, a marker of the ethylene-mediated defense pathway, was increased by 1.9-fold, 9 h after Trichokonin treatment (Fig. 4b). NtCOI1, required for JA response in tobacco, was also induced by Trichokonin treatment, the expression of which RVX-208 was increased by 1.8-fold after a 6-h treatment. These results suggested the involvement of multiple defense pathways in Trichokonins-induced tobacco resistance against TMV. Several peptaibols isolated from Trichoderma spp. have been reported to have antimicrobial activity against Gram-positive bacterial and fungal phytopathogens (Daniel & Filho, 2007). Peptaivirins A and B from Sepedonium spp. are the only two peptaibols known to have antiviral activity against TMV, with an inhibitory effect of 74% and 79%, respectively, at concentration of 10 μg mL−1 (Yun et al., 2000). The Trichokonins isolated from T. pseudokoningii SMF2 have been shown to exhibit antimicrobial activity against a range of Gram-positive bacterial and fungal phytopathogens with a concentration of 20 μg mL−1in vitro (Song et al., 2006).

6% (n = 30 517) Eighty-three per cent (n = 25 243) of respondent

6% (n = 30 517). Eighty-three per cent (n = 25 243) of respondents were working as a pharmacist and were therefore eligible to complete the work/life balance statements. The results reported here relate to 12 364 individuals who had full data for the work/life balance scale and the demographic and work variables. Findings indicate that age, ethnicity, having caring responsibilities, sector of practice, hours of work and type of job are significant predictors of work/life balance problems. Pharmacy employers and buy LEE011 government should recognise the changing demographic characteristics of the profession and consider what support might be available to the workforce

to help alleviate work/life balance problems being experienced by certain groups

of pharmacists. “
“This study evaluated the barriers and facilitators that were experienced as pharmacists were integrated into 23 existing primary care teams located in urban and rural communities in Saskatchewan, Canada. Qualitative design using data from one-on-one telephone interviews with pharmacists, mTOR inhibitor physicians and nurse practitioners from the 23 teams that integrated a new pharmacist role. Four researchers from varied backgrounds used thematic analysis of the interview transcripts to determine key themes. The research team met on multiple occasions to agree on the key themes and received written feedback from an external auditor and two of the original interviewees. Seven key themes emerged describing the barriers and facilitators that the teams experienced during the pharmacist integration: (1) relationships, trust and respect; (2) pharmacist role definition; (3) orientation and support; (4) pharmacist personality and professional experience; (5) pharmacist presence and visibility; (6) resources and funding; and (7) value of the pharmacist role. Teams from urban and

rural communities experienced some of these challenges in unique ways. Primary care teams that integrated a pharmacist experienced several common barriers and facilitators. The negative impact of these barriers can be mitigated Selleckchem Lumacaftor with effective planning and support that is individualized for the type of community where the team is located. “
“Objectives  To investigate older patient, physician and pharmacist perspectives about the role of pharmacists in pharmacist-patient interactions. Methods  Eight focus-group discussions were held in senior centres, community pharmacies and primary care physician offices. Participants were 42 patients aged 63 years and older, 17 primary care physicians and 13 community pharmacists. Qualitative analysis of the focus-group discussions was performed. Key findings  Participants in all focus groups indicated that pharmacists are a good resource for basic information about medications. Physicians appreciated pharmacists’ ability to identify drug interactions, yet did not comment on other specific aspects related to patient education and care.

Simple indexes are easy to implement and cost-effective However,

Simple indexes are easy to implement and cost-effective. However, the diagnostic yield of these indexes is lower in HIV/HCV-coinfected patients [3,4]. In particular, the diagnosis of cirrhosis cannot be established confidently [3]. TE seems a promising technique for use in HIV/HCV-coinfected patients [5,6]. However, the high rate of classification errors produced by use of a single cut-off value precludes its application

for establishing the absence of fibrosis and detecting mild fibrosis in coinfected patients [5,6]. Use of two cut-off values to detect and exclude significant fibrosis improves the diagnostic accuracy of TE in HIV/HCV selleck kinase inhibitor coinfection, but leaves a substantial proportion of patients unclassified [7]. In addition,

TE is not widely available because of its high cost, and regulatory issues are a barrier to access to TE in some countries. Fibrosis is a wound-healing response characterized by increased fibrogenesis and fibrolysis, both of which may produce increased levels of circulating extracellular matrix components or their fragments [8,9]. Matrix metalloproteinases and their inhibitors, tissue inhibitors of metalloproteinases, are enzymes controlling matrix degradation [8]. Matrix metalloproteinase 2 (MMP-2) is expressed in liver injury and degrades normal extracellular matrix. As a consequence, normal low-density basement membrane is replaced Tamoxifen in vivo with fibril-forming matrix that is deleterious to hepatocyte function [8]. Tissue inhibitor of metalloproteinase 1 (TIMP-1) inactivates proteases and can have antiapoptotic effects on hepatic stellate cells, thus leading to an increased pool of fibrogenic cells [8]. Serum levels of MMP-2 and TIMP-1 may therefore correlate with liver fibrosis. Multi-component tests have been developed, some of which include measurements of metalloproteinases and their inhibitors, in various combinations to predict fibrosis in HCV infection [9–14]. However, there is little information on the diagnostic yield of these serum biomarkers in HIV/HCV-coinfected patients [15–18]. Noninvasive

diagnosis Nintedanib (BIBF 1120) of liver fibrosis needs to be improved in HIV/HCV coinfection. Simple serum indexes can spare liver biopsy in up to half of patients [19]. Serum tests which include measurements of extracellular matrix markers (e.g. the SHASTA index) or which are entirely based on markers of fibrogenesis also leave a significant proportion of patients without a definitive diagnosis [15,16]. TE is less accurate in identifying patients with less advanced fibrosis [5,6]. Against this background, we examined the utility of serum MMP-2 and TIMP-1 in combination with routinely available data to predict liver fibrosis in HIV/HCV-coinfected patients. This was a retrospective cross-sectional study carried out in the Infectious Diseases Unit of Hospital Universitario de Valme, Seville, southern Spain, from November 1999 to December 2006.

P and JL were recipients of a graduate fellowship provided by

P. and J.L. were recipients of a graduate fellowship provided by the MEST through the Brain Korea 21 Project. “
“Host immune pressure and associated immune evasion of pathogenic bacteria are key features of host-pathogen co-evolution. A previous study showed that human T-cell epitopes Torin 1 cost of Mycobacterium tuberculosis are evolutionarily hyperconserved and thus it was deduced that M. tuberculosis lacks antigenic variation and immune evasion. Here, we

selected 173 clinical M. tuberculosis complex (MTBC) isolates from China, amplified the genes encoding Rv2945c and Rv0309, and compared the sequences. The results showed that genetic diversity existed in these two genes among the MTBC strains and two single nucleotide polymorphisms (SNPs) presented higher polymorphisms. Antigen Rv2945c harbored a higher number of amino acid substitutions of its T-cell epitopes, which may reflect ongoing

immune evasion. In addition, the high dN/dS value of Rv0309 suggested antigen Rv0309 might be involved in diversifying selection to evade Ganetespib cost host immunity. Finally, a small group of strains were identified based on the genetic diversity of these two genes, which might indicate that they interact differently with human T cells compared with other strains. “
“Farnesyl pyrophosphate (FPP) is utilized for many cellular processes, including the production of dolichols, ubiquinone (CoQ), sterols, farnesylated heme A and prenylated proteins. This lipid synthesized Aprepitant by FPP synthetase (ERG20) becomes attached to target proteins by the prenyltransferases, CDC43/RAM2 and RAM1/RAM2 complexes after the formation of the C15 and C20 units, respectively. Defects in protein prenylation as a result of inhibiting these enzyme complexes lead to pleiotropic effects in all eukaryotes. In this study, using Candida glabrata conditional mutants, the importance of the ERG20 and RAM2 genes for growth using both in vivo and in vitro assays was assessed by placing the RAM2 and

ERG20 genes under the control of a regulatable promoter. Repression of RAM2 gene expression revealed growth defects under both conditions. However, repression of ERG20 gene expression did not impair fungal growth in a mouse host, but did result in growth defects on laboratory media. Thus, FPP synthase is not required for survival in an infected mouse, but the RAM2-encoded prenyltransferase was critical for growth under both conditions. This study strongly suggests that inhibitors of prenyltransferase may be promising antifungals. Farnesyl pyrophosphate (FPP), produced by the isoprenoid pathway, serves as a precursor of metabolites including sterols, dolicols and ubiquinones and as a substrate for protein prenylation required for, among other processes, signal transduction and membrane anchoring (Fig. 1). Specifically, FPP, sterol biosynthesis and protein prenylation are prominent drug targets for the development of a wide range of inhibitors (Gelb et al., 2006; Kuranda et al.

Similarly, 6-hydroxydopamine-induced chronic dopaminergic denerva

Similarly, 6-hydroxydopamine-induced chronic dopaminergic denervation induced a significant increase in expression of AT1, AT2 and p47phox, which decreased with L-dopa administration. A significant reduction in expression of AT1 mRNA was also observed after administration of dopamine to cultures of microglial cells. Transgenic rats with very low levels of brain AII showed increased AT1, decreased p47 phox and no changes in AT2 expression, whereas mice deficient in AT1 exhibited a decrease in the expression of p47 phox and AT2. The administration of relatively high doses of AII (100 nm) decreased the expression of AT1, and the increased expression of AT2 and p47phox in primary mesencephalic cultures.

The results reveal an important interaction between the dopaminergic and local renin–angiotensin system in the basal ganglia, which may be a major factor selleck chemicals llc in the progression of Parkinson’s disease. “
“Thermoregulation enables adaptation to different ambient temperatures. A complex network of central autonomic centres may be involved. In contrast to the brainstem, the role of the cortex has not been clearly evaluated. This study was therefore designed to address cerebral function during a whole thermoregulatory cycle (cold, neutral and warm stimulation)

using 18-fluordeoxyglucose-PET (FDG-PET). Sympathetic activation parameters were co-registered. Ten healthy male volunteers were examined three times on three different days in a water-perfused whole-body suit. After Protirelin a baseline period (32°C), temperature was either decreased to 7°C (cold), increased to 50°C (warm) or kept constant (32°C, this website neutral), thereafter the PET examination was performed. Cerebral glucose metabolism was increased in infrapontine brainstem and cerebellar hemispheres during cooling and warming, each compared with neutral temperature. Simultaneously, FDG uptake decreased in the bilateral

anterior/mid-cingulate cortex during warming, and in the right insula during cooling and warming. Conjunction analyses revealed that right insular deactivation and brainstem activation appeared both during cold and warm stimulation. Metabolic connectivity analyses revealed positive correlations between the cortical activations, and negative correlations between these cortical areas and brainstem/cerebellar regions. Heart rate changes negatively correlated with glucose metabolism in the anterior cingulate cortex and in the middle frontal gyrus/dorsolateral prefrontal cortex, and changes of sweating with glucose metabolism in the posterior cingulate cortex. In summary, these results suggest that the cerebral cortex exerts an inhibitory control on autonomic centres located in the brainstem or cerebellum. These findings may represent reasonable explanations for sympathetic hyperactivity, which occurs, for example, after hemispheric stroke. “
“The molecular mechanisms leading to neurodegeneration in Parkinson’s disease remain elusive.

, 2010)

, 2010). Tamoxifen cost However, only two sequences of small plasmids from Arthrobacter species are deposited in GenBank database. The plasmid pA3 (AJ131246) is 2205 bp in length and harbours five hypothetical open reading frames (ORF). The second plasmid (pRE117-2, FQ311476) is 8528 bp in length, and 13 ORFs are predicted, two of them encode putative mobilization proteins (Monnet et al., 2010). Recently, Miteva et al. (2008) have described the cryptic plasmid p54 (1950 bp), which harbours seven ORFs, few of which sharing similarities with proteins of known function. However, the nucleotide sequence is not publicly available. To date, a few vectors for the bacteria of Arthrobacter genus have been

created. Two hybrid plasmids buy ICG-001 have been developed using the ori sequence of pCG100 from Corynebacterium glutamicum (Shaw & Hartley, 1988; Sandu et al., 2005) and pBL100 from Brevibacterium lactofermentum (Shaw and Hartley, 1988).

One vector has been constructed on the basis of pULRS8 from Brevibacterium lactofermentum (Morikawa et al., 1994). The pART2 and pART3 vectors can be applied for both constitutive and nicotine-inducible gene expression as well as for promoter screening by GFP fusion (Sandu et al., 2005) or production of MalE-fused hybrid proteins (Kolkenbrock & Fetzner, 2010). All above-mentioned E. coli–Arthrobacter shuttle vectors are developed from cryptic plasmids of phylogenetically related species. Recently, the hybrid vector Thiamet G pSVJ21 has been constructed

based on the cryptic plasmid p54 from Arthrobacter sp. (Miteva et al., 2008). This paper reports on characterization of a small cryptic plasmid pPRH (5.0 kb) from Arthrobacter rhombi PRH1 strain and describes the pPRH-derived hybrid vectors, which replicates in both Arthrobacter and Rhodococcus species as well as in E. coli. One of the vectors has been successfully applied for functional screening of 2-hydroxypyridine catabolism encoding genes from Arthrobacter sp. PY22, using a nonconventional host. The bacterial strains and plasmids are listed in Table 1. Arthrobacter and Rhodococcus spp. strains were cultivated at 30 °C on nutrient agar (NA) (Oxoid) plates or in nutrient broth (Oxoid) aerobically. When necessary, antibiotics were added to the media: ampicillin (50 μg mL−1), chloramphenicol (10–20 μg mL−1), kanamycin (40–60 μg mL−1 and tetracycline (10–40 μg mL−1). Cloning and DNA manipulations were performed as described by Maniatis et al. (1982). Plasmid DNA from Rhodococcus and Arthrobacter cells was isolated by alkaline lysis method following the incubation with lysozyme (10 mg mL−1) for 30 min. Escherichia coli and Arthrobacter (Rhodococcus) cells were prepared for electroporation by the method of Sharma & Schimke (1996) and Gartemann & Eichenlaub (2001), respectively. A restriction analysis of the pPRH plasmid was carried out using single and double digestions. The DNA fragments were subcloned in pTZ57R.

4 kb);

4 kb); http://www.selleckchem.com/products/PLX-4720.html EF650850 (MTT1-BS07 2.4 kb); EF650851 (MTT1-BS07 2.7 kb); EF650852 (MTT1-A15 2.4 kb) and EF650853 (MTT1-WS 2.7 kb). Previously deposited sequences

are available under accession numbers DQ010168–DQ010174. Sequences were aligned using clustalw (Thompson et al., 1994). prosite was used to find motifs and membrane-spanning domains in the purported proteins (http://www.expasy.org/prosite/). Binding sites in the promoters were analysed using siteseer (Boardman et al., 2003). We have reported previously that antimycin A strongly inhibits the growth of lager strain A15 on a solid medium with maltotriose, but has less effect on the growth of lager strain WS34/70 (Dietvorst et al., 2005). As shown in Fig. 1, lager strain BS07 was also inhibited in its growth on maltotriose

in the presence of antimycin A, but to a smaller extent than lager strain A15. A fourth lager yeast strain, BS01, shows a similar growth profile as strain WS34/70. For growth on maltose as a carbon source, the effect of antimycin A was less and about the same for all four strains (Fig. 1). To investigate the presence of MTT1-like and/or selleck chemicals llc MAL31-like genes in the lager yeast strains A15, WS34/70, BS01 and BS07, PCRs were performed using specific primer combinations MAL31-fw – MAL31-rv and Mty1-fw – Mty1-rv, respectively (Table 1 and Fig. 2). These primers discriminate between MTT1- and MAL31-like genes. Using these primers, we showed that all four lager strains contain both MAL31 and MTT1 genes (data not shown). To isolate MAL31 Amrubicin and MTT1 genes from the four lager yeast strains, independent PCRs were performed using the universal primers ‘MAL31Xba’and ‘MAL31BamH’. This PCR amplifies the sequences between 542 or 836 bp upstream and 26 bp downstream of the open reading frame (ORFs) and yielded both 2.4- and 2.7-kb products as reported previously for strains A15 and

WS34/70 (Dietvorst et al., 2005). The PCR products were inserted into the pCR-TOPO vector and independent clones were isolated and characterized by PCR with the MTT1-specific pimers Mty1-fw and Mty1-rv and the MAL31-specific primers Mal31-fw and Mal31-rv (Table 1 and Fig. 2). From strains WS34/70 and BS07, both 2.4- and 2.7-kb versions of the MAL31 and MTT1 genes were isolated, whereas the 2.7-kb version of MTT1 was not found in strains A15 and BS01 (Table 2). To further study the role of these genes in maltotriose metabolism, the various MAL31 and MTT1 genes were recloned into the multicopy vector pRUL409(KanMX). A15 transformants containing these constructs were tested for their ability to start growing rapidly on maltotriose in the presence of antimycin A. For each strain, at least two independent isolates of both the 2.4- and the 2.7-kb versions of both the MAL31 and the MTT1 genes were tested in this manner, except for the 2.7-kb MTT1 versions from strains BS01 and A15, which were not found. Transformants carrying the 2.