B.-B. “
“The clone Escherichia coli O25 ST131, typically producing extended-spectrum beta-lactamases (ESBLs), has spread globally and became the dominant type among extraintestinal isolates at many parts of the world. However, the reasons behind the emergence and success of this clone are only partially understood. We compared the core

type genes by PCR of ESBL-producing and ESBL-nonproducing strains isolated from urinary tract infections in the United Arab Emirates and found a surprisingly high frequency of the K-12 core type (44.6%) among members of the former group, while in the latter one, it was as low (3.7%), as reported earlier. The high figure was almost entirely attributable to the presence of members of the clone O25 ST131 among ESBL producers. Strains from Bleomycin the same clone isolated in Europe also carried the K-12 core type genes. Sequencing selleck chemical the entire core operon of an O25 ST131 isolate revealed a high level of similarity to known K-12 core gene sequences and an almost complete identity with a recently sequenced

non-O25 ST131 fecal isolate. The exact chemical structure and whether and how this unusual core type contributed to the sudden emergence of ST131 require further investigations. In Escherichia coli, the core oligosaccharide (OS) part of the lipopolysaccharide (LPS) molecule occurs in five different types: R1–4 and K-12, respectively

(Muller-Loennies et al., 2007). The core has a crucial role in maintaining the structure of the cell wall, although to what extent and how its specific type affects the colonizing capacity or the virulence of a pathogen remains to be elucidated. Nevertheless, earlier studies consistently found a highly disproportional distribution of these core types among commensal and clinical E. coli isolates (Gibb et al., 1992; Appelmelk et al., 1994; Amor et al., Vitamin B12 2000; Gibbs et al., 2004). Among strains recovered from extraintestinal infections, the frequency of R1 core type reached 61.0–81.0%, while that of the K-12 type was found the least or the second least common (2.2–5.6%) (Gibb et al., 1992; Appelmelk et al., 1994; Amor et al., 2000). These frequencies were well reflected by the distribution of core-type-specific antibodies in the population (Gibbs et al., 2004). In the past decade, the spread of extended-spectrum beta-lactamase (ESBL)-producing E. coli strains considerably altered the epidemiology and treatment options of extraintestinal infections (Woodford et al., 2011; Van der Bij et al., 2012). A significant percentage of these isolates belong to a limited number of clones, some considerably differing in their panel of virulence factors from those described earlier (Totsika et al., 2011; Van der Bij et al., 2012).

In this study, although p24 antigen ELISA testing was able to detect the same HIV-positive cases

identified by the nucleic acid technique, previous studies suggested that p24 antigen testing could identify from 79 to 90% of acute infections [10]. Thus, the p24 antigen ELISA may be an option for improving early detection of HIV infections only where access to nucleic acid-based detection methods is limited. In conclusion, the results of this study suggest that the algorithm for early diagnosis of acute Ponatinib mw HIV infections should include individual nucleic acid detection in MSM with HIV-negative WB with discordant results in the screening assays, as well as in those with HIV-indeterminate WB. An accurate early diagnosis of

acute HIV infection may benefit patients by permitting clinical interventions, which can limit viral spread by decreasing viral loads and thus reducing the risk of transmission. The authors thank Fundación Alberto J. Roemmers (Buenos Aires, Argentina) for financial support and Siemens Argentina for the donation of reagents. The authors also thank Mr Sergio Mazzini for revision of the manuscript. “
“Antiretroviral therapy during pregnancy is recommended to reduce the risk of mother-to-child transmission of HIV and for maternal care management. Physiological changes during pregnancy can affect pharmacokinetics, potentially altering pharmacological activity. We therefore evaluated the pharmacokinetics of twice-daily (bid) darunavir in HIV-1-infected pregnant women. HIV-1-infected pregnant women receiving an antiretroviral regimen containing darunavir/ritonavir 600/100 mg bid were enrolled in this study. MK-1775 in vivo Total and unbound darunavir and total ritonavir plasma concentrations were not obtained over 12 h during

the second and third trimesters and postpartum. Total darunavir and ritonavir plasma concentrations were determined using a validated high-performance liquid chromatography tandem mass spectrometry assay and unbound darunavir was determined using 14C-darunavir-fortified plasma. Pharmacokinetic parameters were derived using noncompartmental analysis. Data were available for 14 women. The area under the plasma concentration–time curve from 0 to 12 h (AUC12h) for total darunavir was 17–24% lower during pregnancy than postpartum. The AUC12h for unbound darunavir was minimally reduced during pregnancy vs. postpartum. The minimum plasma concentration (Cmin) of total and unbound darunavir was on average 43–86% and 10–14% higher, respectively, during pregnancy vs. postpartum. The antiviral response (< 50 HIV-1 RNA copies/mL) was 33% at baseline and increased to 73–90% during treatment; the percentage CD4 count increased over time. One serious adverse event was reported (increased transaminase). All 12 infants born to women remaining in the study at delivery were HIV-1-negative; four of these infants were premature. Total darunavir exposure decreased during pregnancy.

Pujol for advice. This work was supported in part by the project with reference AGL2011-30461-C02-02 by the Ministerio de Ciencia e Innovación (Spain). Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“The Gram-negative anaerobe Dichelobacter nodosus is the causative agent of footrot in sheep. Different strains of D. nodosus cause disease of differing severities, ranging from benign to virulent.

Virulent strains have greater twitching motility and secrete proteases that are more thermostable than those secreted by benign strains. We have identified polynucleotide phosphorylase (PNPase) as a putative virulence regulator and have proposed that PNPase expression is modulated by the adjacent integration of genetic elements. In this study, RO4929097 clinical trial we compared PNPase activity in three virulent and four benign strains of D. nodosus and found that PNPase activity is lower in virulent selleck screening library strains. We disrupted the pnpA gene in three benign D. nodosus strains and two virulent strains and showed that deletion of the S1 domain of PNPase reduced catalytic activity. In all but one case,

deletion of the PNPase S1 domain had no effect on the thermostability of extracellular proteases. However, this deletion resulted in an increase in twitching motility in benign, but not in virulent strains. Reconstruction of the pnpA gene in two mutant benign strains reduced twitching motility to the parental level. These results support the hypothesis that PNPase is a virulence repressor in benign strains of D. nodosus. Footrot is a mixed bacterial infection Dimethyl sulfoxide of the hooves of sheep, goats and deer that leads to lameness. The Gram-negative anaerobic bacterium Dichelobacter nodosus is the principal causative agent (Beveridge, 1941). Different strains of D. nodosus cause disease of differing severities, ranging from benign to virulent. The extracellular proteases secreted by virulent

strains are more thermostable than proteases secreted by benign strains (Depiazzi & Richards, 1979). Virulent strains also have greater twitching motility, generated by polar type IV fimbriae, than benign strains (Depiazzi & Richards, 1985), and twitching motility is essential for virulence (Kennan et al., 2001; Han et al., 2008). Comparative analysis of DNA from virulent and benign strains has led to the identification of a series of genetic elements that integrate into the D. nodosus chromosome. These include the intA (Katz et al., 1991, 1992, 1994; Cheetham et al., 1995; Billington et al., 1996), intB (Bloomfield et al., 1997), intC (Bloomfield et al., 1997) and intD elements (Tanjung et al., 2009), each of which contains an integrase gene. A fifth integrated element, the virulence-related locus, vrl (Katz et al., 1991; Haring et al., 1995; Billington et al., 1999), lacks an integrase gene.

The primary endpoints of the present substudy were changes from baseline in plasma levels of interleukin-6 (IL-6), interleukin-8 (IL-8), monocyte

chemotactic protein-1 (MCP-1), soluble vascular cell adhesion molecule-1 (sVCAM-1), soluble CD40 ligand (sCD40L), soluble P-selectin (sP-selectin) and tissue plasminogen activator (t-PA) in Buparlisib cell line the two arms at months 12, 24 and 36. Secondary endpoints were correlations of these biomarkers with viral load and plasma lipids. At baseline and at months 12, 24 and 36, venous blood samples were obtained after an overnight fast and frozen at −70°C until analysis. IL-6, IL-8, MCP-1, sVCAM-1, sCD40L, sP-selectin and t-PA levels were measured in cell supernatants by a multiplex cytometric bead-based assay (Human Cardiovascular selleck 7plex FlowCytomix Multiplex; Bender Medsystems GmbH, Vienna, Austria), using an EPICS-XL-MCL

flow cytometer (Beckman Coulter, IZASA, Barcelona, Spain), following the manufacturer’s protocol. In brief, 25 μL of the 7 mixed beads and biotin-conjugate mixture was mixed with 25 μL of the standards or samples provided and incubated in the dark for 2 h at room temperature. Samples were then washed, 25 μL of streptavidin-phycoerythrin (PE) solution was added, and incubation was carried out for a further 1 h. After the second incubation, samples were washed and resuspended in 300 μL of assay buffer. The EPICS-XL-MCL flow cytometer was calibrated with set-up beads and 300 events were acquired for each factor and each sample, respectively. Individual analyte concentrations were indicated by their fluorescence intensities (FL-2) and

computed with the respective standard reference curve and FlowCytomixPro 2.2 software. Standard curves were determined for each biomarker from a range of 27 pg/mL to 40 000 ng/mL. According to the manufacturer, the detection limits of the assay are 0.9 pg/mL for IL-6, 7.9 pg/mL for IL-8, 53.0 ng/mL for sP-selectin, 8.0 pg/mL for t-PA, 11.0 pg/mL for MCP-1, 0.4 ng/mL for sVCAM-1, and 50.0 pg/mL for sCD40L. Total-c, HDL-c and triglycerides were measured using standard methods. LDL-c was calculated using the Friedewald equation. Peripheral blood CD4 T-cell count was determined by flow cytometry and plasma viral load by real-time polymerase chain reaction (PCR) (Abbott RealTime HIV-1; PRKACG Abbott Laboratories, Abbott Park, IL). Quantitative variables are expressed as the median and interquartile range (IQR). Before the statistical analysis, the normality of distributions and homogeneity of variances were tested. sP-selectin and sCD40L were log10-transformed because of high distribution variability. The two-sample t-test or Mann–Whitney U-test was used to compare continuous variables between arms. Qualitative variables were compared using the χ2 or Fisher exact test. Baseline and follow-up values in each arm were compared with the paired t-test or Wilcoxon signed rank test.

No significant differences in sociodemographic variables between the sites were found. The mean age was 43 years (range 21–73 years) and the subjects had been aware of their HIV infection for a mean of 9.6 years (range 1–26 years). Table 1 shows further sample characteristics. For the sample of patients recruited in Essen, 822 patients attending the clinic selleck chemicals fulfilled the criteria for participation during the observation period. Of these, 409 were formally asked to participate in the study. Of these 409 subjects, 245 (59.9%) participated in the study and 138 (33.7%) refused

to participate. In addition, 26 subjects (6.4%) were excluded (11 subjects did not fulfil the inclusion criteria, 10 had incomplete data, three took part twice, and two interrupted the examination). In total, 49.7% of all possible subjects participated. Comparable recruitment figures were found in Bochum, where, in total, 49.8% of possible subjects participated. In total, 88.5% of the subjects had been sexually active in the past 12 months. One-quarter of the participants reported one male partner (25.6%) during this period and another quarter reported two to five male partners (25.2%). Furthermore, 12.8% had sexual contact with six to 10 men, 17.8% with 11 to 50 men and 7.9% with more than 50 different

male partners. The majority (53.2%) indicated a frequency of sexual activity ranging from several times per months to several times per week. More than half of all participants (57.2%) reported unprotected sexual contact. Unprotected LEE011 nmr insertive anal intercourse was reported by 34.6% and unprotected receptive anal intercourse by 32.9% during the last 12 months. For the description of substance use, we differentiated between current and lifetime substance use (never, less than three and more than three times per week). For the lifetime prevalence, the category ‘less than three times ever’ was added. For alcohol use, we differentiated between any alcohol use and alcohol use until drunkenness. If

the report of the frequency of substance use suggested the possibility of a substance-related disorder, the criteria of the ICD-10 (10th edition of the International Dichloromethane dehalogenase Statistical Classification of Diseases and Related Health Problems published by the World Health Organization) for addiction or harmful use were applied. There was a remarkably high prevalence of current use of amyl nitrite (26.4%), amphetamines (7.2%), dissociative drugs such as ketamine (2.6%), and erectile dysfunction medication (11.4%). The prevalence of currently manifest substance addiction was 4.5% for cannabis, 3.9% for alcohol and 0.2% for amphetamines (for detailed results, see Tables 2 and 3). We found significant correlations between the use several substances and sexual risk behaviour. The most obvious effect was found for amyl nitrite and cannabis.

Motor sequence acquisition through practice involves at least two distinct, yet interrelated processes in the nervous system: online processes leading to improvements in skill performance during practice, and offline processes that lead to either stabilization of the skill performance over time (memory stabilization) or improvement in skill performance between training sessions (offline learning) (Robertson & Cohen, 2006). Sequence learning is implemented by a network of cortical and subcortical structures that are engaged during practice as well as after

practice (Doyon et al., 1997, 2003; Karni et al., 1998; Robertson et al., 2001; Press selleck chemicals et al., 2005). Acquisition of serial behavior may involve implicit or explicit learning. Implicit sequence learning refers to improvement in performance of the sequence without overt information about the elements of a sequence. In contrast, explicit sequence learning is accompanied by explicit conscious recollection of each element and its order in the sequence (Squire, 1986; Vidoni & Boyd, 2007; Robertson, 2009). There are multiple differences in the explicit and implicit memory systems, including the neural substrates that implement implicit and explicit learning. Using positron emission tomography, Honda and colleagues demonstrated that anatomically distinct networks

were associated with implicit and explicit sequence learning. Implicit sequence learning was primarily associated with activity in the contralateral sensory and M1 (Pascual-Leone et al., 1994). In contrast, when learners developed explicit knowledge about the practiced sequence, see more activation

in the dorsal premotor cortex (PMd), dorsolateral prefrontal cortex and supplementary motor area correlated strongly with conscious recall of the sequence (Honda et al., Buspirone HCl 1998; Vidoni & Boyd, 2007; Robertson, 2009). Implicit and explicit memory systems are complex and often compete to mediate task performance. Learning a word-list (explicit memory task) immediately after implicit motor sequence practice enhanced learning of the motor sequence (Brown & Robertson, 2007a). This suggested that sequence-related information in the explicit memory system probably competes with implicit memory system, and blocking that sequence-related explicit information (with a word-list) allows the implicit memory system to maximize motor learning. Here we investigated the neural basis of competition between the implicit and explicit systems during implicit motor sequence learning. We used anodal transcranial direct current stimulation (AtDCS) to modulate the excitability of distinct neural structures known to be engaged in implicit (primary motor cortex, M1) and explicit (PMd) memory systems during implicit motor sequence practice. The effect of AtDCS on M1 and PMd was assessed with online and offline changes in motor performance.

Concerning the colour, the fungus B. cinerea can attack the grape berry and introduce the oxidative enzyme laccase into the berry and hence into grape juice. Laccase targets phenolics such as the red colour compounds in red wine and oxidizes them into brown-coloured compounds. Furthermore, the association of B. cinerea with other, less visible, fungi frequently leads to the development of organoleptic defects in grapes and sometimes in wines (La Guerche et al., 2006). The strategy most widely adopted by winegrowers to reduce the impact of grey selleckchem mould is the systematic application of chemical fungicides, based on a preset calendar that takes into account the phenological growth

stages of the grapevine. This reduction policy will have an impact on Botrytis resistance to fungicides (Leroux, 2004) and on the environment. Indeed, the contamination of agricultural soils with

inorganic (Cu-based) and organic pesticides (including their residues) presents a major environmental and toxicological click here concern (Komárek et al., 2010). Although there are alternative methods to synthetic fungicides, such as the application of antagonistic microorganisms and the application of natural antimicrobial substances, it is essential to monitor the disease development and particularly the concentration of fungal spores. Indeed, monitoring disease development will allow better disease management, and will reduce cost and improve grape quality. Spores can be identified and quantified by light microscopy (Aylor, 1998; Hunter et al., 1999). However, this is not straightforward. Indeed, it is a time-consuming technique that needs expertise for the accurate identification of spores. Antibody immunoassays have been used for the early detection of B. cinerea (Kennedy et al., 2000). However, taking into account the low sensitivity and the limited dynamic range of the method, it is not well adapted for quantification, although it can be used to confirm the nature of the agent (Suarez et al., 2005). Molecular techniques for the identification of spores have

already been published (West et al., 2008), most of which are based on detection by standard PCR methods (Zhou et al., 2000; Calderon et al., 2002; Chew et al., 2006). However, under these conditions, quantification is not Fossariinae precise. One way to assess for the presence of specific spores more accurately and to avoid some of the problems that accompany the other methodologies is real-time quantitative PCR (qPCR). Numerous quantitative assays utilizing real-time PCR have been developed to specifically detect microbial targets in many types of samples, including, but not limited to, moulds (Alaei et al., 2009; Carisse et al., 2009; Luo et al., 2010). Advantages of utilizing qPCR for spore enumeration over classic culture-based methods include its enhanced specificity and reduced processing time, leading to quicker results. Cadle-Davidson (2008) reported a qPCR method based on Taqman chemistry for monitoring B.

In our experience this arrangement does not compromise the recording quality of the silicon probe. Experiments with the microbial light-sensitive protein Clamydomonas reinhardtii ChR2 (Nagel et al., 2003; Boyden et al., 2005; Li et al., 2005; Ishizuka

et al., 2006; Han & Boyden, 2007; Zhang et al., 2007a and b) were carried out in rats. To obtain neuronal expression of ChR2 in the hippocampus, the CA1 region of 3-week-old animals was injected with the adenoassociated virus (AAV) encoding ChR2–green-fluorescent protein (GFP) fusion protein. Briefly, the fusion protein was cloned into an AAV cassette containing check details the mouse synapsin promoter, a woodchuck post-transcriptional regulatory element (WPRE), SV40 polyadenylation sequence and two inverted terminal repeats.

Viral particles were assembled using a modified helper-free system (Stratagene, La Jolla, CA, USA) as a serotype 2/5 (rep/cap genes) AAV, and harvested and purified over sequential cesium chloride gradients as previously described (Grieger et al., 2006). The injections were performed stereotaxically under isofluorane anesthesia through a burr-hole above the dorsal hippocampus, using a glass pipette (10 μm tip size) connected to a microinjector (Nanoject II; Drummond Scientific Comp., Broomall, PA, USA). Volumes of 45 nL (undiluted stock, minimum 1011 NU7441 cell line viral particles per mL) were injected every 300 μm between depths

of 2.0 and 2.6 mm below dura, at three locations along the CA1 septotemporal axis (2.8–4.2 mm anterior to bregma and 2.5–2.8 mm lateral). Ten weeks Thiamine-diphosphate kinase after the virus injection, the rats were trained to run on an elevated figure-eight maze, built by the assembly of modular aluminum segments. Water rewards were delivered at two corners of the maze through water ports controlled by valves (no. 003-0130-900; Parker Pneutronics). Custom-made motorized doors forced the animals to take the right turns at the two intersections of the maze. Light-beam sensing switches (no. 65845K7; McMaster) detecting the animal’s passages at some locations were used for the automatic triggers of valves, doors and laser for ChR2 activation. Twelve weeks after the virus injection, the rats were prepared for chronic recordings. The general surgical procedures have been described (Fujisawa et al., 2008; Royer et al., 2010). Briefly, the prepared optrode assembly was attached to a micromanipulator. Under isofluorane anesthesia, two small watch-screws were driven into the bone above the cerebellum to serve as reference and ground electrodes. After enlarging the hole used for the virus injection, the dura mater was removed. The probe was positioned so that its shanks avoided puncturing large veins and inserted 1 mm into the brain.

, 2010). To confirm that this advantage applies to Purkinje cells, we sought to molecularly perturb their early developmental processes by IUE. The ataxic mouse mutant staggerer is caused by a deletion in the gene encoding RORα1 (Sidman et al., 1962; Hamilton et al.,

1996). As RORα1 lacking PI3K Inhibitor Library the putative ligand-binding domain (RORα1DN) serves as a dominant-negative mutant in cultured muscle cells (Lau et al., 1999, 2004) (Fig. 5A), we introduced two plasmids, pCAG-RORα1DN-HA, in which HA-tagged RORα1DN was placed under the CAG promoter, and pCAG-EGFP, into Purkinje cells by IUE at E11.5. The mice were fixed at P9, and sagittal sections at the vermis were immunostained for calbindin and HA to visualize Purkinje cells and RORα1DN, respectively.

Confocal microscopy showed that almost all the control calbindin-positive Purkinje cells expressing EGFP had single primary dendrites (96.2%, 102 of 106 cells; Fig. 5B and C). By contrast, only half of the calbindin-positive Purkinje cells expressing EGFP and RORα1DN-HA had a single primary dendrite (49.5%, 50 of 101 cells; P < 0.0001 vs. control, χ2 test), and PD-0332991 supplier the remaining cells had from two to five primitive dendrites (Fig. 5B and C). Furthermore, while all the control Purkinje cells expressing EGFP were arranged in a monolayer together with non-transfected Purkinje cells, a small number of Purkinje cells expressing RORα1DN-HA (six of 101) were mislocalized to the granular layer (Fig. 5B, arrowheads). These phenotypes observed in Purkinje cells expressing RORα1DN-HA were reminiscent of those observed in staggerer Purkinje cells (Soha & Herrup, 1995; Nakagawa et al., 1998). These results clearly indicate that certain

staggerer phenotypes can be mimicked by the IUE-mediated expression of dominant-negative RORα1 in single Purkinje cells during early development. Although IUE has several advantages as a method for transferring genes into neurons in vivo, it has never been applied next to cerebellar Purkinje cells, key neurons for regulating cerebellar functions. In the present study, we showed that Purkinje cell progenitors at E11.5 could be most efficiently and preferentially transfected by IUE, by properly adjusting the angle and direction of the electrodes (Fig. 1). Electrophysiological analyses indicated that the electroporated Purkinje cells maintained normal membrane properties, synaptic responses and synaptic plasticity at P28 (Fig. 2). We also showed that simultaneous expression of three different fluorescent proteins (Fig. 4) and expression of a large gene (Bassoon; Fig. S4) could be successfully achieved by IUE in Purkinje cells. In addition, by using three plasmids encoding the L7 promoter and an inducible Cre/Lox system, we could achieve temporal and Purkinje-cell-specific transgene expression (Fig. 3).

, 2010). To confirm that this advantage applies to Purkinje cells, we sought to molecularly perturb their early developmental processes by IUE. The ataxic mouse mutant staggerer is caused by a deletion in the gene encoding RORα1 (Sidman et al., 1962; Hamilton et al.,

1996). As RORα1 lacking this website the putative ligand-binding domain (RORα1DN) serves as a dominant-negative mutant in cultured muscle cells (Lau et al., 1999, 2004) (Fig. 5A), we introduced two plasmids, pCAG-RORα1DN-HA, in which HA-tagged RORα1DN was placed under the CAG promoter, and pCAG-EGFP, into Purkinje cells by IUE at E11.5. The mice were fixed at P9, and sagittal sections at the vermis were immunostained for calbindin and HA to visualize Purkinje cells and RORα1DN, respectively.

Confocal microscopy showed that almost all the control calbindin-positive Purkinje cells expressing EGFP had single primary dendrites (96.2%, 102 of 106 cells; Fig. 5B and C). By contrast, only half of the calbindin-positive Purkinje cells expressing EGFP and RORα1DN-HA had a single primary dendrite (49.5%, 50 of 101 cells; P < 0.0001 vs. control, χ2 test), and Palbociclib concentration the remaining cells had from two to five primitive dendrites (Fig. 5B and C). Furthermore, while all the control Purkinje cells expressing EGFP were arranged in a monolayer together with non-transfected Purkinje cells, a small number of Purkinje cells expressing RORα1DN-HA (six of 101) were mislocalized to the granular layer (Fig. 5B, arrowheads). These phenotypes observed in Purkinje cells expressing RORα1DN-HA were reminiscent of those observed in staggerer Purkinje cells (Soha & Herrup, 1995; Nakagawa et al., 1998). These results clearly indicate that certain

staggerer phenotypes can be mimicked by the IUE-mediated expression of dominant-negative RORα1 in single Purkinje cells during early development. Although IUE has several advantages as a method for transferring genes into neurons in vivo, it has never been applied Tangeritin to cerebellar Purkinje cells, key neurons for regulating cerebellar functions. In the present study, we showed that Purkinje cell progenitors at E11.5 could be most efficiently and preferentially transfected by IUE, by properly adjusting the angle and direction of the electrodes (Fig. 1). Electrophysiological analyses indicated that the electroporated Purkinje cells maintained normal membrane properties, synaptic responses and synaptic plasticity at P28 (Fig. 2). We also showed that simultaneous expression of three different fluorescent proteins (Fig. 4) and expression of a large gene (Bassoon; Fig. S4) could be successfully achieved by IUE in Purkinje cells. In addition, by using three plasmids encoding the L7 promoter and an inducible Cre/Lox system, we could achieve temporal and Purkinje-cell-specific transgene expression (Fig. 3).