Bacterial genomic DNA was prepared using the cetyltrimethylammonium bromide method (Ausubel et al., 1993). The purified DNA was quantified using the Nano Quant Infinite M200 spectrophotometer (Tecan, Männedorf, Proteases inhibitor Switzerland) at a wavelength of 260 nm. Primers and fluorescent dye-labeled TaqMan MGB probes were designed based on the nucleotide sequences that corresponded to the cps gene of S. pneumoniae (GenBank accession number NC_011072) obtained from SSH using the primer express 3.0 program (Applied Biosystems, Foster City, CA). The primer set cpsA-348F (5′-GCTGTTTTAGCAGATAGTGAGATCGA-3′) and cpsA-415R (5′-TCCCAGTCGGTGCTGTCA-3′)

defined an amplicon of 67 base pairs (bp). A carboxyfluorescein (FAM)-labeled probe cpsA-TaqMan FAM (5′-FAM-AATGTTACGCAACTGACGAG-MGBNFQ1-3′) was used for detection.

Standard curves and minimal limit of detection were generated by plotting the cycle threshold values (CT) of the qPCR performed on a dilution series of purified DNA from S. pneumoniae KCTC 5080T cells (107–1 CFU mL−1) against the log input cells mL−1. Streptococcus pneumoniae concentrations were calculated using a viable cell plate count method. Serial 10-fold dilutions of RXDX-106 cell line the cultures were inoculated on BHI agar (Difco Laboratories). The plates were subsequently incubated at 37 °C for 16 h, and cultural counts (in CFU) were determined in triplicate. The primer and probe concentrations for each of the three assays were optimized, and, in accordance with the experimentally optimized concentrations, 250 nM cpsA-specific primers and 150 nM cpsA-specific probes were used for all subsequent experiments. DNA was amplified with the 7300 real-time PCR system (Applied Biosystems) using the following cycling parameters: 95 °C for 10 min, followed by 50 cycles of 95 °C for 15 s

and 60 °C for 1 min. Amplification data were analyzed using sds software (7300 Real-time PCR System Sequence Detection Software v1.31, Applied Rho Biosystems). A specimen was considered positive if two of the three triplicates yielded a positive result within the <50-cycle cutoff. Conventional PCR-based methods have been developed to differentiate S. pneumoniae strains from the closely related viridans group streptococci. However, the lateral gene transfers that occur among the viridans group streptococci limit this differentiation, making it difficult to diagnosis S. pneumoniae in the human environment. In fact, some pneumococcal virulence genes of S. pneumoniae have been detected in S. mitis or S. oralis isolates (Whatmore et al., 2000; Verhelst et al., 2003; Seki et al., 2005). The cpsA gene was identified as a novel genomic marker specific to S. pneumoniae using the SSH technique, which allows accurate discrimination of S. pneumoniae from the closely related viridans group streptococci. In our study, a qPCR technique targeting the cpsA gene was developed to detect and enumerate the human pathogen, S.

As well, it would probably be valuable to explore whether such neurofunctional reorganization also occurs in aging animal models. Such studies would be important in order to distinguish between the engagements of such normal age-related phenomena and those phenomena linked to a neurodegenerative process. Abbreviations BA selleckchem Brodmann area CRUNCH compensation-related utilization of neural circuits hypothesis fMRI functional magnetic resonance imaging HAROL Dhemispheric asymmetry reduction in older adults PASA posterior–anterior shift in aging STAC scaffolding theory of aging and cognition VF verbal fluency “
“Using a transgenic mouse (Mus musculus) in which nestin-expressing progenitors

are labeled with enhanced green fluorescent protein, we previously characterized the expression of excitatory amino acid transporter 2 (GltI) and excitatory amino acid transporter 1 (Glast) on early neural progenitors in vivo. To address their functional role in this cell population, we manipulated their expression in P7 neurospheres isolated from the dentate gyrus. We observed that knockdown

of GltI or Glast was associated with decreased bromodeoxyuridine incorporation and neurosphere formation. Moreover, we determined that both glutamate transporters regulated progenitor proliferation in a calcium-dependent and metabotropic glutamate receptor-dependent manner. To address the relevance of this in vivo, we utilized models mafosfamide of acquired brain selleck inhibitor injury, which are known to induce hippocampal neurogenesis. We observed that GltI and Glast were specifically upregulated in progenitors following brain injury, and that this increased expression was maintained for many weeks. Additionally, we found that recurrently injured animals with increased

expression of glutamate transporters within the progenitor population were resistant to subsequent injury-induced proliferation. These findings demonstrate that GltI and Glast negatively regulate calcium-dependent proliferation in vitro and that their upregulation after injury is associated with decreased proliferation after brain trauma. “
“Reward sensitivity, or the tendency to engage in motivated approach behavior in the presence of rewarding stimuli, may be a contributory factor for vulnerability to disinhibitory behaviors. Although evidence exists for a reward sensitivity-related increased response in reward brain areas (i.e. nucleus accumbens or midbrain) during the processing of reward cues, it is unknown how this trait modulates brain connectivity, specifically the crucial coupling between the nucleus accumbens, the midbrain, and other reward-related brain areas, including the medial orbitofrontal cortex and the amygdala. Here, we analysed the relationship between effective connectivity and personality in response to anticipatory reward cues.

Patients regularly followed at the Department of Infectious Diseases, San Raffaele Scientific Institute, Milan, Italy, with known HIV-1 infection since before 1988, no previous diagnosis of DM, and available HCV and HBV serology data were contacted between February and June 2008 and asked: (i) to undergo a complete physical examination,

including blood pressure MK-2206 cell line and anthropometry; (ii) to complete a questionnaire to evaluate their family history of DM, their current smoking history, and their use of lipid-lowering agents and antihypertensive medications; (iii) to provide a fasting blood sample for the measurement of glucose, insulin, total cholesterol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, and triglycerides; and (iv) to undergo an OGTT on a different day within a month of the first blood sample. All of the study participants gave their informed consent to take part in the study. Blood samples were collected after an overnight fast (defined as at least 12 h), which was always rigorously verified. All of the parameters were tested by means of routine standard procedures (Diagnostic Unit, San Raffaele Scientific Institute, Laboraf). The homeostatic model assessment for insulin resistance (HOMA-IR) index was calculated according to Matthews et al. as [fasting glucose (mg/dL) × baseline insulin (mIU/L)]/405 [28]. A standard 75-g OGTT was used to assess

2-h post-load glucose levels. Glucose values were interpreted on the basis of the criteria recommended by the American Diabetes Association [1]: FPG<100 mg/dL (<5.6 mmol/L)=normal fasting glucose; FPG 100–125 mg/dL (5.6–6.9 mmol/L)=impaired Selleckchem Daporinad fasting glucose; FPG≥126 (≥7 mmol/L)=provisional diagnosis of diabetes; 2-h post-load glucose <140 mg/dL (<7.8 mmol/L) =normal glucose tolerance; 2-h post-load glucose 140–199 mg/dL (7.8–11 mmol/L)=IGT; 2-h post-load glucose ≥200 mg/dL (≥11.1 mmol/L)=provisional diagnosis of diabetes. The subjects' family history of DM was evaluated by means of a self-administered questionnaire and was considered positive if at least one first-degree relative was/had been diabetic. Waist circumference was classified as normal

or abnormal on the basis of Methane monooxygenase the American Heart Association/National Heart, Lung, and Blood Institute (AHA/NHLBI) criteria (abnormal for males: ≥102 cm; abnormal for females: ≥88 cm) [29]. Sitting blood pressure was determined using a sphygmomanometer after a >5-min rest. Coinfection with hepatitis B virus (HBV) and hepatitis C virus (HCV) was defined as the presence of HBV surface antigen and HCV antibodies, respectively. The characteristics of the patients are described using median values and quartiles (Q1–Q3) or frequencies and percentages (%), as appropriate. The differences between subjects with IGT or DM and those with normal OGTT results were assessed for significance using Wilcoxon’s two-sample rank sum test for nonparametric data.

, 2005). Other plasmids frequently used in BF638R are also difficult or impossible to introduce into BF 9343 (data not shown). In general, more efficient transposon mutagenesis is achieved by prior modification of plasmid carrying the transposon by the host of interest. We developed an improved system for transposon mutagenesis in BF using the EZ::TN5 system. Previous attempts to mutagenize BF by transposons have been hindered

by either vector integration and/or multiple insertions (Shoemaker et al., 1986; Chen et al., 2000a). Also, those methods often used labor-intensive filter mating techniques to introduce the DNA. The method described here has several advantages: (1) transposons can be introduced into BF by electroporation, (2) all insertion events are independent, (3) no vector delivery

AZD8055 supplier system is required and vector cointegration can be completely avoided, and (4) no suicide vector or native inducible promoters to drive transposase expression are needed. We found that the transposon inserts evenly across the chromosome. Also, analysis of the insertion points of the EZ::TN5 transposon indicates that although there is some sequence context preferred of insertion by Tn5, the insertion is sufficiently random for its effective use in construction a library of transposon mutants (Shevchenko et al., 2002). EZ::TN5 transposon mutagenesis also provides flexibility for subsequent identification of the transposon-disrupted gene. For example, if the genome sequence is not available Navitoclax datasheet for Epothilone B (EPO906, Patupilone) the organism of interest, the genes adjacent to the mutated gene can be retrieved and identified by rescue cloning and sequencing. On the other hand, if the genome sequence is available, the mutated gene can be amplified by SRP-PCR and identified by genomic means and large numbers of mutants can be easily screened. Prior passage of the transposon vector in related strains increases downstream efficiency of transposon mutagenesis. This system provides a useful genetic tool that will facilitate deeper understanding of

the pathogenic mechanisms of this important human commensal/pathogen. This research is based upon work supported in part by the Department of Veterans Affairs, Veterans Health Administration, Office of Research and Development, Biomedical Laboratory Research and Development and in part by the NIAID (NIH) Grant Number 1R56AI083649-01A2. We would like to thank Drs Elizabeth Tenorio and Yi Wen for their helpful comments and advice regarding mutant identification and Southern Blots, respectively. “
“Rapid detection of yeast contamination is important in the food industry. We have developed loop-mediated isothermal amplification (LAMP) assays to detect the emerging opportunistic pathogenic yeasts: Candida albicans, Candida glabrata, Candida tropicalis, the Candida parapsilosis group, Trichosporon asahii, and Trichosporon mucoides.

, 2005). Other plasmids frequently used in BF638R are also difficult or impossible to introduce into BF 9343 (data not shown). In general, more efficient transposon mutagenesis is achieved by prior modification of plasmid carrying the transposon by the host of interest. We developed an improved system for transposon mutagenesis in BF using the EZ::TN5 system. Previous attempts to mutagenize BF by transposons have been hindered

by either vector integration and/or multiple insertions (Shoemaker et al., 1986; Chen et al., 2000a). Also, those methods often used labor-intensive filter mating techniques to introduce the DNA. The method described here has several advantages: (1) transposons can be introduced into BF by electroporation, (2) all insertion events are independent, (3) no vector delivery

RG7422 molecular weight system is required and vector cointegration can be completely avoided, and (4) no suicide vector or native inducible promoters to drive transposase expression are needed. We found that the transposon inserts evenly across the chromosome. Also, analysis of the insertion points of the EZ::TN5 transposon indicates that although there is some sequence context preferred of insertion by Tn5, the insertion is sufficiently random for its effective use in construction a library of transposon mutants (Shevchenko et al., 2002). EZ::TN5 transposon mutagenesis also provides flexibility for subsequent identification of the transposon-disrupted gene. For example, if the genome sequence is not available this website for Lck the organism of interest, the genes adjacent to the mutated gene can be retrieved and identified by rescue cloning and sequencing. On the other hand, if the genome sequence is available, the mutated gene can be amplified by SRP-PCR and identified by genomic means and large numbers of mutants can be easily screened. Prior passage of the transposon vector in related strains increases downstream efficiency of transposon mutagenesis. This system provides a useful genetic tool that will facilitate deeper understanding of

the pathogenic mechanisms of this important human commensal/pathogen. This research is based upon work supported in part by the Department of Veterans Affairs, Veterans Health Administration, Office of Research and Development, Biomedical Laboratory Research and Development and in part by the NIAID (NIH) Grant Number 1R56AI083649-01A2. We would like to thank Drs Elizabeth Tenorio and Yi Wen for their helpful comments and advice regarding mutant identification and Southern Blots, respectively. “
“Rapid detection of yeast contamination is important in the food industry. We have developed loop-mediated isothermal amplification (LAMP) assays to detect the emerging opportunistic pathogenic yeasts: Candida albicans, Candida glabrata, Candida tropicalis, the Candida parapsilosis group, Trichosporon asahii, and Trichosporon mucoides.

The volume was calculated according to the procedure of Cavalieri, and variability within groups was assessed via the coefficient of error (CE). CEs for all analyses were = 0.10. To quantify graft innervation, TH+ sections 240 μm apart were analysed using the Space Balls estimator program (StereoInvestigator, MicroBrightfield, Williston, VT, USA) to obtain an unbiased estimate of TH+ neurite density in the striatum. Fiber density analyses were conducted

in 4–6 selleck compound serial sections. Contours were drawn for three fields of view at the lateral border of the graft at 4×, and neurites that crossed the borders of the hemispheric probe were counted at 60× with oil immersion. Neurite density was calculated as neurite length/volume. The volume was calculated according to the procedure of Cavalieri, and variability within groups was assessed via the CE. CEs for all analyses were = 0.10. A modified bootstrapping method was used for the analysis of behaviors that had extensive temporal data. This approach involved the inclusion of re-sampled data from the following

time-point http://www.selleckchem.com/products/bmn-673.html groupings: ‘pre-graft maturation’ time-point (weeks −2, 0 and 2 post-grafting); ‘early post-grafting’ time-point (weeks 4 and 6 post-grafting); ‘mid post-grafting’ time-point (weeks 8 and 10 post-grafting); and a ‘late post-grafting’ time-point (weeks 18 and 20 post-grafting). A two-way repeated-measures analysis of variance (anova) was performed for each behavior, to assess the effects of treatment, time, and treatment by before time interaction. Significant differences of main effects were determined using Bonferroni post hoc analyses. Differences in spine density, TH+ cell counts and TH+ fiber densities were determined using one-way anovas followed by Tukey’s

post hoc analyses. Analysis of Golgi-treated striatal tissue showed a > 40% reduction in spine density on dendrites both distal and proximal to the cell bodies of MSNs in the dopamine-depleted striatum compared with controls (control: distal = 10.52 ± 0.85 spines per 10 μm, proximal = 12.32 ± 0.79 spines per 10 μm; 6-OHDA-treated: distal = 5.57 ± 0.83 spines per 10 μm, proximal = 6.78 ± 0.88 spines per 10 μm). This loss was protected against in parkinsonian rats receiving nimodipine pellets at both distal and proximal sites, with nimodipine-treated rats showing no significant difference from intact controls (6-OHDA + nimodipine: distal = 9.02 ± 0.41 spines per 10 μm, P = 0.39; proximal = 10.78 ± 0.58 spines per 10 μm, P = 0.42) but differing significantly from parkinsonian rats receiving vehicle pellets (distal: F2,12 = 12.15, P = 0.01; proximal: F2,12 = 13.54, P = 0.007; Fig. 2). Both dopamine-grafted groups showed significantly reduced rotational behavior when compared with sham-grafted controls (early post-graft: dopamine-grafted = 0.38 ± 0.18 rotations per min, dopamine-grafted + nimodipine = 0.42 ± 0.23 rotations per min, sham-grafted = 3.08 ± 1.

Raltegravir was generally well tolerated over 96 weeks of treatment in HIV-infected patients see more with and without HBV and/or HCV coinfection. The incidence of hepatobiliary adverse events ranged from 0 to 3% in patients with HBV or HCV and from 3 to 4% in those without HBV or HCV coinfection. Grade 2–4

liver enzyme elevations were observed more frequently in patients with HIV and hepatitis coinfection than in HIV-monoinfected patients, but this difference was noted in both the raltegravir and control groups. These results are consistent with two recent reports. Rachlis et al. [17] found that, among patients receiving darunavir with low-dose ritonavir in the POWER 1 and GPCR Compound Library in vitro 3 studies, patients with HBV or HCV coinfection had a higher incidence of ALT and AST elevations than those without coinfection. Vispo et al. [18] found that liver enzyme elevations occurred more frequently in HIV/HCV-coinfected patients than in HIV-monoinfected patients (P<0.001) across four antiretroviral drug classes, and that liver enzyme elevations were less frequent in patients receiving raltegravir or maraviroc than in those receiving nonnucleoside reverse transcriptase inhibitors or protease inhibitors. With regard to efficacy, we found that the antiretroviral

and immunological effects of raltegravir were similar in patients with HIV and HBV/HCV coinfection and those with HIV infection only. The studies included in these analyses were not designed to compare 3-mercaptopyruvate sulfurtransferase treatment effects in patient subgroups based on hepatitis coinfection

status. In the BENCHMRK studies, there may be relevant differences in important baseline characteristics between the subgroups because patients were not stratified by hepatitis coinfection status. In addition, the method for defining HCV infection in the BENCHMRK studies may represent a bias, as patients with HCV antibodies consist of patients with chronic HCV disease as well as successfully resolved HCV infection, which could lead to lower hepatotoxicity rates. Despite these limitations, the results of the current analyses suggest that raltegravir is generally well tolerated and efficacious for the treatment of HIV infection in patients with HBV and/or HCV coinfection, and is therefore an appropriate therapeutic alternative for these patients. Merck Sharp & Dohme Corp., a subsidiary of Merck & Co. Inc., provided financial support for the studies included in this report. “
“Long-term antibody responses to 23-valent pneumococcal polysaccharide vaccine (PPV) among HIV-infected patients receiving highly active antiretroviral therapy (HAART) are rarely investigated. Antibody responses to three pneumococcal capsular polysaccharides [Pneumococcal polysaccharide (PPS) 14, 19F and 23F] were assessed among 169 HIV-infected patients who received HAART and 23-valent PPV.

, 1999; Macomber et al., 2007). Copper, in either the Cu(I) or Cu(II) states, has strong affinity for sulfhydryl groups, and the binding of copper to thiol or nitrogen-containing groups in proteins could inhibit protein function (Gerba & Thurman, 1989; Kershaw et al., 2005). However, copper is more toxic to bacterial buy Buparlisib cells under anaerobic conditions, where there is a greater proportion of Cu(I) (Beswick et al., 1976; Outten et al., 2001). Early work on copper suggested that Cu(I) is more toxic owing to increased binding to amino acids and nucleosides (Cramp, 1967). Cu(I) displaces the iron in iron–sulfur clusters

and binds to the thiol groups in important metabolic enzymes (Macomber & Imlay, 2009). To avoid the toxicity exerted by copper, bacteria utilize intricate mechanisms to reduce free intracellular concentrations of the metal (Osman & Cavet, 2008). Although selleck there are distinct differences between copper and silver in their role in and effects on biological systems, these metals share very similar chemical and ligand-binding properties. Cu(I) and Ag(I) belong to the group

of soft Lewis acids that have high polarizability and form bonds with nitrogen- and sulfur-containing molecules, which are soft Lewis bases (Housecroft & Sharpe, 2005). Silver can actively compete for copper sites in biomolecules, thus disrupting their function and key interactions (Dibrov et al., 2002). It has been observed that systems that aid in copper homeostasis can also actively detoxify silver (Rensing et al., 2000; Stoyanov et al., 2003). Regulatory control of metal concentrations

in living organisms is vital to prevent cellular damage. Owing to the toxic nature of the metals, bacteria have 4��8C developed sophisticated mechanisms conferring silver and copper resistance (Grass & Rensing, 2001b; Rensing & Grass, 2003; Grass et al., 2011). In Escherichia coli, the Cue and the Cus systems detoxify/remove excess silver and copper from the cells. The Cue response system consists of CopA, a P-type ATPase that exports intracellular Cu(I) into the periplasm (Rensing et al., 2000), and CueO, a periplasmic multicopper oxidase that oxidizes Cu(I) to Cu(II) (Grass & Rensing, 2001a). The Cus response system consists of the chemiosmotic CusCFBA efflux system (Grass & Rensing, 2001b; Franke et al., 2003). The Cus system is activated when the Cue system is overwhelmed with copper or under anaerobic conditions, when the oxidase CueO is inactive (Outten et al., 2001). The Cus system is particularly important to confer periplasmic Ag(I) tolerance to the cell, as CueO is inhibited by Ag(I) (Singh et al., 2011).

There are also indirect estimates of the dominance of fungal denitrification in alkaline soils after the application of bacterial or fungal

inhibitors (Castaldi & Smith, 1998; Laughlin & Stevens, 2002; Crenshaw et al., 2008). Fungi are thought to contribute to N2O production through nitrite or nitrate reduction, as denitrification or codenitrification (Bollag & Tung, 1972; Shoun et al., 1992; Tanimoto et al., 1992), under low oxygen (O2) conditions, for example ‘initially selleckchem aerobic’ culture vessels (Zhou et al., 2001). Fungal denitrification occurs in the fungal mitochondria (Kobayashi et al., 1996), whereas bacterial denitrification is restricted to the cell membrane. However, the universality of this trait within all fungal groups is unknown. The symbiotic mutualistic ectomycorrhizal fungi dominate the microbial biomass in acidic temperate and boreal Crizotinib order forest soils (Smith & Read, 2008).

These fungi form symbiotic associations with tree roots (e.g. pine, birch, poplar): in return for carbon (C) derived from host-plant photosynthesis, the fungi forage and acquire nutrients for their host via the extensive fungal mycelial network. Although fungi, in general terms, were proposed as a source of N2O in acidic forest soils (Bleakley & Tiedje, 1982), the ability of the ectomycorrhizal fungal group to produce N2O or their contribution to soil N2O fluxes remains unknown. Ectomycorrhizal fungi can grow on certain nitrogen (N) sources, proteins, amino acids, ammonium and nitrate (Finlay et al., 1992); however, the N reduction pathway in ectomycorrhizal fungi is poorly understood compared with other fungal groups. For example, the presence of the nitrate reductase enzyme in 68 ectomycorrhizal fungal species Cyclin-dependent kinase 3 was only recently confirmed (Nygren et al., 2008). Here, we provide the first evidence of the ability of two ectomycorrhizal fungi, Paxillus involutus (Batsch) Fr. and Tylospora fibrillosa (Burt.) Donk, which are highly competitive when inorganic N concentrations are high (Brandrud, 1995; Carfrae et al.,

2006), to produce N2O through nitrate reduction under low O2 conditions. N2O production by these fungi was compared with that of the known fungal denitrifier, F. lichenicola [CBS 483.96; Centraalbureau voor Schimmelcultures (CBS), the Netherlands] previously known as Cylindrocarpon tonkinense IFO 30561 (Shoun et al., 1992; Usuda et al., 1995; Watsuji et al., 2003). The production of N2O was examined in fungi P. involutus 8 (Batsch) Fr. (from Sheffield University), T. fibrillosa F23 3AT (Burt.) Donk (isolated from Sitka spruce root tips) and F. lichenicola CBS 483.96, under aseptic pure culture conditions. The growth medium was modified Melin–Norkrans liquid medium (Marx, 1969) with glucose, ammonium and malt extract omitted and the pH adjusted to 5.6.

5a). In the control strain, approximately 70% of hyphae contained stained Spk 30 min after the initial staining (Fig. 5b and c), which increased to 90% after 60 min (Fig. 5b). In contrast, in the

aipA-overexpressing strain, approximately 35% of hyphae with stained Spk were observed 30 min after the staining (Fig. 5b and c), which only increased to 50% after 60 min (Fig. 5b). Notably, the mutant aipA-overexpressing strains showed nearly identical Spk staining as that of the control AG-014699 price strain (Fig. 5b and c). Taken together, these results suggest that the endocytic recycling of FM4-64 to Spk is both defective and delayed in the aipA-overexpressing strain. However, because the aipA-overexpressing strain also displayed impaired growth, it is possible that the Spk was not present in certain hyphae,

and thus, the relative rate of endocytic recycling was not substantially delayed in this strain. To exclude this possibility, we calculated the half-time required for Spk staining with FM4-64 for each of the strains (Fig. 5d). The half-time for staining in the aipA-overexpressing strain was clearly longer than that in the control and mutant aipA-overexpressing strains, indicating that the find protocol aipA-overexpressing strain has defects with respect to endocytic recycling; this delay could be caused by the defect of endocytosis, that of trafficking of vesicles to Spk, or both. We also confirmed that there was no significant difference between the control and the ΔaipA strains in this analysis (data not shown). In this study, we discovered a putative

AAA ATPase, AipA, as a binding partner of AoAbp1 by YTH screening. Although the ΔaipA strain did not display growth or endocytic defects, the aipA-overexpressing strain showed impaired growth, abnormal hyphal morphology, and a deficiency in the endocytic recycling of FM4-64, whereas the mutant aipA-overexpressing strains did not. The subsequent localization and functional analyses using the aipA-overexpressing strain suggested that AipA negatively functions second in endocytic recycling at the tip region of A. oryzae. There seems to be one AipA ortholog in filamentous fungi and two in yeasts. Both Sap1p and Yta6p, S. cerevisiae AipA orthologs, are putative AAA ATPases, but their molecular function is unknown. Sap1p was found by the YTH analysis as a binding protein with Sin1p, a transcriptional repressor (Liberzon et al., 1996). Yta6p is one of 12 YTA family proteins and is localized at the cortex in mother cells, but not in daughter cells (Schnall et al., 1994; Beach & Bloom, 2001). Single disruptants of either SAP1 or YTA6 are viable and no remarkable phenotypic alteration has been reported.