In particular, rapid methods that can be performed in the field, minimizing the delay between sampling and diagnosis, could reduce the spread of this pathogen. The most common method for rapid detection of P. sojae, which is found globally, was NVP-LDE225 in vivo developed based on conventional polymerase chain reaction (PCR)(Wang et al., 2006). However, this method might not be suitable for developing countries because of the high-tech equipment required, elaborate and complicated assay procedures, expensive reagents, time requirements, and the frequency of false-positives. Therefore, there is a growing demand for simple and economical molecular tests. In this study, we developed an alternative amplification method that can

be used in Rucaparib concentration the field to detect P. sojae in the absence of a thermal cycler.

Loop-mediated isothermal amplification (LAMP) of DNA is a novel technique that uses a set of four or six primers and the strand displacement activity of Bst DNA polymerase to amplify DNA with high specificity under isothermal conditions (Notomi et al., 2000). LAMP products can be monitored by measuring the increased turbidity (due to the production of large amounts of magnesium pyrophosphate) in real time, and visualized by gel electrophoresis or by adding hydroxynaphthol blue (HNB) prior to amplification (Ma et al., 2010). The simplicity of the LAMP method, which does not require a thermal cycler, makes it suitable for field testing. Although this method has been applied in the field of microbiology for detection and identification of bacteria (Pan et al., 2011), viruses (Parida et al., 2004) and fungi (Niessen & Vogel, 2010), the technique has not been applied to P. sojae. LAMP is simple once the appropriate primers have been designed based on the target gene (Notomi et al., 2000). PCR-based and quantitative real-time PCR-based methods for the detection of P. sojae have CHIR-99021 in vitro been described based on ribosomal gene sequences (Wang et al., 2006). However, rRNA gene sequences from closely related species are highly

conserved, limiting the development of species-specific detection primers. As a result, a new target with high specificity and efficiency is required to distinguish between closely related species for rapid detection of P. sojae. With the development of Phytophthora genomics and numerous molecular targets, the identification efficiency of Phytophthora species has increased significantly. We identified a new P. sojae identifiable target, named. A3aPro is a 300-bp deletion element in the upstream (1.5 kb in the promoter region) of the avirulence gene Avr3a in P. sojae Race 7, as compared with Race 2 and 12 (Supporting Information, Fig. S1). Further bioinformatics analysis showed it is a transposon-like element whose high-identity copies were commonly distributed in the sequenced P. sojae genome but absent in non-P. sojae species. We acquired the A3aPro sequence in the P.

In MSM, awareness was also associated with having a university degree, the degree of interaction with gay culture, number of partners, and use of the internet as the main way of meeting partners. nPEP awareness in the studied population was unacceptably low. The promotion of its availability should be made a major objective of prevention programmes, as a complementary measure to condom use. “
“In the UK, free HIV care is provided through dedicated HIV clinics. Using LY2109761 molecular weight the national cohort of men who have sex with men (MSM) with diagnosed HIV infection and estimates of the number of undiagnosed men, we assessed whether high retention in HIV care and

treatment coverage is sufficient to reduce HIV transmission. Antiretroviral therapy (ART) uptake and viral load distribution among diagnosed men were analysed by treatment status and CD4 count for the period between 2006 and 2010. A multi-parameter evidence synthesis click here (MPES) method was used to estimate the size of the undiagnosed population. The viral load distribution among newly diagnosed untreated men was applied to the undiagnosed population. Infectivity was defined as a viral load > 1500 HIV-1 RNA copies/mL. Between 2006 and 2010, ART coverage among

all HIV-infected MSM (diagnosed and undiagnosed) increased from 49 to 60%, while the proportion of infectious men fell from 47 to 35%. Over the same period, the number of all HIV-infected MSM increased from 30 000 to 40 100 and the number of infectious MSM remained stable at 14 000. Of the 14 000 infectious MSM in 2010, 62% were until undiagnosed, 33% were diagnosed but untreated, and 5% received ART. Extending ART to all diagnosed HIV-infected MSM with CD4 counts < 500 cells/μL in 2010 would have reduced the overall proportion of infectious men from 35 to 29% and halving the proportion who were undiagnosed would further have reduced this to 21%. High ART coverage in the UK has

reduced the infectivity of the HIV-diagnosed population. However, the effectiveness of treatment as prevention will be limited unless the undiagnosed population is reduced through frequent HIV testing and consistent condom use. “
“The aim of the study was to describe pregnancies in HIV-infected teenagers. A review of the case notes of HIV-infected pregnant teenagers aged 13–19 years from 12 London hospitals was carried out for the period 2000–2007. There were 67 pregnancies in 58 young women, of whom one was known to have acquired HIV vertically. The overall mother-to-child transmission (MTCT) rate of HIV was 1.5% (one of 66). There were 66 live births. Median ages at HIV diagnosis and conception were 17 and 18 years, respectively. Sixty-three per cent of women were diagnosed with HIV infection through routine antenatal screening. Eighty-two per cent of pregnancies (41 of 50) were unplanned, with 65% of women (26 of 40) using no contraception. Forty-three per cent of the women (20 of 46) had a past history of a sexually transmitted infection (STI).

These activations

were prevented by blocking mGluR2/3 with LY341459, an mGluR2/3 antagonist. Furthermore, selleck chemicals blocking ERK, PI3K and NFκB signaling pathways with U0126, LY294002 and pyrrolidine dithiocarbamate, respectively, significantly inhibited the mGluR2/3-mediated restorative effects. These results suggest that application of mGluR2/3 agonists after OGD insult can effectively reverse the OGD-reduced expression of GLAST proteins and restore clearance of extracellular glutamate by serially activating ERK/PI3K/NFκB signaling pathways in cultured astrocytes. “
“The learning and memory deficits associated with non-pathological ageing mainly result from alterations to the plasticity of neuronal network dynamics within the hippocampus. In addition to the broad spectrum of changes that affect the morphology and function of hippocampal excitatory circuits in the ageing brain, the impaired activation of the N-methyl-d-aspartate subtype of glutamate receptors (NMDA-R)

is a typical feature, altering the induction and maintenance of long-term potentiation, a major form of synaptic plasticity. In addition to glutamate, Transmembrane Transporters modulator the binding of a co-agonist at the strychnine-insensitive glycine-binding site is required for NMDA-R activation. This review presents recent evidence that: (i) the amino acid d-serine is an endogenous co-agonist of synaptic NMDA-R and necessary for long-term potentiation expression, (ii) reduced d-serine levels in the hippocampus contribute to synaptic plasticity and memory deficits in normal ageing, and Coproporphyrinogen III oxidase (iii) age-related oxidative stress selectively targets hippocampal serine racemase to impact d-serine availability in neuronal networks. These results emphasize the critical role of the hippocampal

d-serine-dependent pathway in changes affecting neuronal network dynamics in physiological ageing that underlie memory deficits. In addition, the central role of serine racemase in these changes opens new perspectives in the search for relevant therapeutic strategies aimed at reducing age-related memory defects. “
“There is great interest in outlining biological factors and behavioral characteristics that either predispose or predict vulnerability to substance use disorders. Response to an inescapable novel environment has been shown to predict a “drug-use-prone” phenotype that is defined by rapid acquisition of cocaine self-administration. Here, we showed that response to novelty can also predict the neurochemical and behavioral effects of acute and repeated cocaine in rats. We used cocaine self-administration under a fixed-ratio 1 schedule followed by fast-scan cyclic voltammetry in brain slices to measure subsecond dopamine (DA) release and uptake parameters in drug-use-prone and -resistant phenotypes.

This analysis showed

that iron, 5′-aminolevulinic acid (ALA) and possibly haem control haem biosynthesis mostly via modulating expression of hemA [coding for 5′-aminolevulinic acid synthase (ALAS)]. A hemA deletion mutant (ΔhemA) was constructed, which showed conditional lethality. Growth of ΔhemA was supported on standard nitrate-containing media with ALA, but not by hemin. Growth of ΔhemA could be sustained in the presence of hemin in combination with ammonium instead of nitrate as N-source. Our results suggest that a branch-off within the haem biosynthesis pathway required for sirohaem synthesis is responsible for lack of growth of ΔhemA in media containing nitrate as sole N-source, because of the requirement of sirohaem for nitrate assimilation, as a cofactor of nitrite reductase. In contrast to http://www.selleckchem.com/products/AZD2281(Olaparib).html the situation in Saccharomyces cerevisiae, cysteine, but not methionine, was found to further improve growth of ΔhemA. These results demonstrate that A. niger can use exogenous hemin for its cellular

processes. They also illustrate important differences in regulation of haem biosynthesis and in the role of haem and sirohaem in A. niger compared to S. cerevisiae. Haem is suggested to be a limiting factor in large-scale production of fungal peroxidases, which require this compound as a co-factor (Andersen et al., 1992; Elrod et al., 1997). Addition of hemin, a Cl-ligand of haem, to culture Selleckchem Epacadostat medium improves this production (Andersen et al., 1992; Elrod et al., 1997; Conesa et al., 2000), but is not suited for industrial applications (Elrod et al., 1997). Also, the mechanisms by which hemin supplementation improves peroxidase production are still unknown. To achieve improved and cost-effective

production of peroxidases by filamentous fungi, knowledge Selleck 5-Fluoracil on haem synthesis and regulation is required as current knowledge is mainly restricted to the first two genes in the pathway (Bradshaw et al., 1993; Elrod et al., 1997, 2000). Haem is an essential molecule for almost every organism owing to its requirement as a cofactor of proteins involved in many primary functions like cellular differentiation and gene regulation (Ferreira et al., 1993; Elrod et al., 1997; Panek & O’Brian, 2002; Hamza, 2006). Its biosynthesis in fungi has been extensively studied in Saccharomyces cerevisiae with mutants available for every step within the pathway (Gollub et al., 1977; Urban-Grimal & Labbe-Bois, 1981; Myers et al., 1987; Kurlandzka et al., 1988; Zagorec et al., 1988; Labbe-Bois, 1990; Keng et al., 1992; Amillet & Labbe-Bois, 1995; Camadro & Labbe, 1996; Hoffman et al., 2003). These mutants can be sustained by supplementing hemin to their growth media or by ergosterol or Tween80 addition to supply for essential unsaturated fatty acids (Gollub et al., 1977).

The complementation is dependent on having a suitable phenotype to screen, and we have made use of the complex phenotype of S. meliloti phaC mutants that includes lack of mucoidy on high carbon ratio media such as YM, absence of fluorescence on Nile red-containing media, and reduced growth on polyhydroxyalkanaote cycle intermediates (Aneja et al., 2004). We should also be able to use this strategy

to isolate other polyhydroxyalkanaote synthesis genes such as phaA and phaB from metagenomic libraries. We anticipate the use of this method for the construction of diverse collections of genes encoding polyhydroxyalkanaote synthesis enzymes that might be useful for the optimization and improvement of industrial polyhydroxyalkanaote production through pathway engineering. As more polyhydroxyalkanaote synthase genes Enzalutamide cost are isolated from metagenomic libraries using these methods, it will be Birinapant research buy interesting to see the full range of genes that can be captured. This work was supported by a Natural Sciences and Engineering Research Council of Canada Strategic Projects grant (T.C.C). Fig. S1. Maximum-likelihood tree inferred from coding DNA sequences of polyhydroxyalkanaote synthases listed in Table S1. Fig. S2. Maximum-likelihood

tree inferred from protein sequences of polyhydroxyalkanaote synthases listed in Table S1. Table S1. Organism names and GenBank accession numbers of related polyhydroxyalkanaote synthases. 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. “
“Clostridial cellulosomes are

cellulolytic complexes that are formed by highly specific interactions between one of the repeated cohesin modules present in the scaffolding protein and a dockerin module of the catalytic components. Although Clostridium thermocellum Xyn11A dockerin Tau-protein kinase has a typical C. thermocellum dockerin sequence, in which two amino acid residues are species specifically conserved within the two segments of the dockerin modules, it can recognize Clostridium josui cohesin modules in a non-species-specific manner. The importance of these two conserved amino acids, which are part of the recognition site of the cohesin and dockerin interaction, was investigated by introducing mutations into the first and/or the second segments of the Xyn11A dockerin. Mutations in the first segment did not affect the interactions between dockerin and C. thermocellum and C. josui cohesins. However, mutations in the second segment prevented binding to cohesin proteins. A second round of mutations within the first segment re-established the affinity for both the C. thermocellum and the C. josui cohesins. However, this was not observed for a ‘conventional’ dockerin, Xyn10C.

The results showed that TMS produced a different effect on subjects’ performance in two separate time windows. When TMS was applied at an early time [160-ms stimulus onset asynchrony

(SOA)], we observed suppression of the Simon effect, resulting from a delay of corresponding trials. When TMS was applied at a late time (220 and 250-ms SOA), we observed an increase in the Simon effect, resulting from a delay of non-corresponding trials. These outcomes revealed that the PMd is involved both in the activation of the spatially triggered response and in response selection during spatial find more conflict. “
“Schematic illustration of an Enriched Environment cage, supplied with shelter, tunnel, wooden ladder, scaffold and ball. For details see the article of Sotnikov et al. (Enriched environment impacts buy RG7204 trimethylthiazoline-induced anxiety-related behavior and immediate early gene expression: critical role of Crhr1. Eur. J. Neurosci., 40, 2691–2700). “
“Cover Illustration: Niche-specific stem/progenitor cells and their neuronal progeny are differentially modulated

by modality-specific sensory input in the adult zebrafish brain. Top image shows a neurogenic niche in a chemosensory region containing proliferating (green) radial glial stem/progenitor cells (magenta). Bottom image shows corresponding ultrastructure. For details see the article of Lindsey et al. (Sensory-specific modulation of adult neurogenesis in sensory structures is associated with the type of stem cell present in the neurogenic niche of the zebrafish brain. Eur. J. Neurosci., 40, 3591–3607). “
“Cover Illustration: An artistic depiction of the neural circuitry hypothesized to underlie avoidance responses in Xenopus laevis tadpoles. Artist: Arseny Khakhalin. For details, see the article by Khakhalin et al. (Excitation and inhibition in recurrent networks mediate collision avoidance in Xenopus tadpoles. Eur. J. Neurosci., 40, 2948–2962). “
“Cover Illustration: An artistic depiction of the neural circuitry

hypothesized to underlie avoidance responses in Xenopus laevis tadpoles. Artist: Arseny Khakhalin. For details, see the article by Khakhalin et al. (Excitation and inhibition in recurrent networks mediate collision others avoidance in Xenopus tadpoles. Eur. J. Neurosci., 40, doi: 10.1111/ejn.12664). “
“Opie et al. (2013) investigated cortical plasticity impairment in the obstructive sleep apnea (OSA) patient. They found OSA patients have both altered corticospinal excitability and, importantly, decreased long-term depression (LTD) in the motor cortex, induced by theta burst-patterned repetitive transcranial magnetic stimulation (rTMS). These exciting findings further elucidate the relationship between apnea and decreased motor skills, and may be extended to study other apnea-related cognitive complications.

Pneumocystis jirovecii is a fungus that causes infection specific to humans [3]. The great majority occur in immunocompromised subjects and

are associated with respiratory symptoms [4]. Current evidence suggests that PCP arises by re-infection from an exogenous source [5]. Evidence for nosocomial transmission exists but is limited [5]. Before the advent of preventative Smad tumor therapy and HAART, PCP occurred in up to 80% of HIV-seropositive individuals with AIDS [6]. In the UK this has declined considerably. Almost 90% of cases occur in HIV-seropositive persons with CD4 T-cell counts <200 cells/μL (or a CD4 T-cell percentage <14%). Other predictive factors for PCP in subjects not receiving effective HAART, include non-adherence to prophylaxis, oral candidiasis, oral hairy leukoplakia, unintentional weight loss, recurrent bacterial pneumonia, previous PCP and a high plasma HIV load [6–12]. The typical presentation of PCP is with exertional dyspnoea, which progresses selleck products over several weeks, malaise and a dry cough. An inability to take a deep breath and fever are often apparent [13]. Rarer presentations include a more rapid onset, haemoptysis and pleuritic chest pain. Purulent sputum production suggests bacterial infection – although this can

be present as a co-pathogen in around one-sixth of cases [14]. Physical examination reveals tachypnoea, normal breath sounds or, less frequently, end-inspiratory crackles. Wheezing and signs of focal consolidation or pleural effusion are less common presentations [13]. Spontaneous or infection-associated learn more pneumothorax in an HIV-seropositive individual should prompt exclusion of PCP [15]. Radiological findings in the chest include perihilar haze, interstitial infiltrates (characteristically

sparing the apices and costo-phrenic angles), pneumatocoeles and pneumothoraces. Upper lobe infiltrates alone have been reported to occur in individuals who are receiving inhaled pentamidine prophylaxis. A normal chest radiograph has been reported to occur in up to 39% of patients and should, therefore, not distract from pursuing the diagnosis of PCP if clinically suspected [16,17]. There are no clinical features specific to PCP. Radiology and nuclear medicine tests are not particularly sensitive or specific [18,19]. Other opportunistic infections may mimic the typical radiological features of PCP [20,21]. Demonstration of a fall in oxygenation between rest and exercise has been validated as a reasonably specific test for PCP in cases with a normal or near-normal chest radiograph who have no previous history of PCP [22], but is not reliable enough to make a diagnosis without confirmatory microbiology.

g. Hickok & Poeppel, 2004; Warren et al., 2005). According to the dual stream model, initial auditory processing in the superior temporal gyrus proceeds via a dorsal stream to the inferior parietal lobule and then to the ventrolateral frontal region for auditory-motor integration, which is necessary for mapping the acoustic speech sounds to articulatory acts. At the same time, a ventral stream is hypothesized to map sounds to meaning in lateral temporal areas. A

recent study (Saur et al., 2008) combined fMRI during two prototypical tasks tapping dorsal (speech repetition) and ventral (language comprehension) streams with diffusion tensor imaging. The authors showed that fibers of the arcuate fasciculus and the superior longitudinal fasciculus are indeed linked to speech repetition and those of the Talazoparib in vivo extreme capsule to language comprehension. A clearer understanding of how different zones of language-related cortex are linked together, using both DTI and RSFC approaches, will have a major impact on our understanding of the neural circuits underlying various aspects of linguistic processing. The primary purpose of the present study was to examine the correspondence between the RSFC of the anterior language production zone, comprising left ventrolateral frontal areas http://www.selleckchem.com/Androgen-Receptor.html 6,

44 and 45, and the findings of a recent autoradiographic tract tracing study that established the anatomical connections between the homologues of these areas and perisylvian parietal and temporal cortex in the macaque (Petrides & Pandya, 2009). As such, we limited our primary analyses to the ventrolateral frontal areas 6, 44 and 45. However, area 47/12 (Petrides, 2005), located on the pars orbitalis, also plays a role in human language processing, particularly in higher level aspects of semantic processing that rely on memory retrieval (Petrides Terminal deoxynucleotidyl transferase & Pandya, 2002). Although beyond the scope of this study, the RSFC related to area 47/12

and its consonance with or differentiation from the RSFC exhibited by surrounding cortical areas, such as area 45, is an issue that should be explored in future studies. Mirror neurons’ were initially described by Rizzolatti et al., (1996) in monkey ventral premotor cortex (Gallese et al., 1996) and later in inferior parietal cortex (Fogassi et al., 2005). The defining characteristic of these neurons is that they discharge during the execution of certain actions, but also during the observation of a similar action performed by another agent (Rizzolatti & Craighero, 2004). For example, if a mirror neuron discharges when the monkey grasps an object, it will also fire when the monkey observes another agent (human experimenter) grasping the same object. Mirror neurons were originally observed in area F5 of the monkey (Gallese et al., 1996; Rizzolatti et al.

However, the trend was not seen at day 14. To date, clinical trials attempting to predict adequate antifungal CNS pharmacokinetics for the treatment of CNS fungal infections have been limited [3]. BAMSG 3-01 demonstrated that fluconazole concentrations in the brain closely paralleled serum levels. The median percentage

of CSF compared with serum fluconazole concentrations for the AmB+Fluc800 and AmB+Fluc400 check details arms were 93.7% and 94.6%, respectively, after 14 days of antifungal treatment. These concentration ratios are consistent with previous results [3,7] but are higher than 70%, which is achieved in the absence of meningeal inflammation [8]. Furthermore, CSerum14 and CCSF14 were found to be highly correlated. Therefore, we can surmise that the steady state of metabolism in both serum and CSF had been achieved. The increased serum concentration in patients receiving fluconazole 800 mg/day compared with those receiving a standard dose of fluconazole over time may be explained by the elimination half-life of fluconazole after zero order kinetics and only 10% of elimination because of the selleckchem metabolism [9]. Fluconazole renal clearance has

been found to be positively correlated with eGFR [9]. In the model for AUCSerum, decreased baseline eGFR was associated with high AUCSerum; however, the impact of baseline eGFR decreased as the dose received increased, suggesting that non-renal elimination pathways may become increasingly important as the fluconazole dose increases. The other subject characteristics, including age and BMI, were not associated with pharmacokinetic parameters (data not shown). Major factors affecting fluconazole pharmacokinetics identified previously included renal insufficiency, ageing and drug–drug interactions from concurrent medication use [2,9]. Of note, increased pharmacokinetic concentration was associated with decreased

day 14 CSF WBC count in BAMSG 3-01. Normally, the CSF WBC Amisulpride count increases as a result of inflammation of the CNS and disruption of the blood–brain barrier. The CSF profiles of HIV- and non-HIV-infected patients are similar for conventional bacterial meningitis, but not cryptococcal meningitis. HIV-infected patients with cryptococcal meningitis are more likely to have a low CSF WBC count and are more likely to have a positive CSF culture [1]. During the course of therapy, risk factors for death or poor clinical outcome for cryptococcal meningitis that have been identified previously included abnormal mental status, high CSF cryptococcal antigen titre, low CSF WBC count, disseminated cryptococcal infection, CSF fungal burden in the CSF and lack of flucytosine treatment [10,11]. While this study was not designed to formally assess the association between pharmacokinetic concentration and cryptococcal meningitis outcome, the findings revealed a tendency of association between high levels of fluconazole and favourable outcomes at days 42 and 70.

Coronal slices containing the DRN were placed for 30 min to 1 h in a vial containing ACSF bubbled with 95% O2–5% CO2 at 37 °C. Thereafter, the slices were kept at room temperature in the same conditions and were transferred one at a time into the recording chamber. For patch-clamp and intracellular experiments, the composition of the ACSF was (in

mm): NaCl, 120; NaHCO3, 25; KCl, 2.5; CaCl2, 2; MgCl2, 2; NaH2PO4, 1.25; and glucose, 10; pH was adjusted to 7.4 with HCl and osmolarity to 300 mOsm with additional glucose. The solution used for extracellular recordings was similar, except that the concentrations of NaCl and KCl were 130 and 3.5 mm, respectively, and no osmolarity adjustment was made. Slices were placed in a recording chamber and continuously superfused with ACSF (at a rate of 2–3 mL/min) Ku-0059436 cell line which was heated to 32 °C using a Thermoclamp (Automate scientific, Berkeley, CA, USA) and a BPS-8 valve control system (ALA scientific, Westbury, NY, USA). Neurons were visualized using an Axioscop 1FS upright microscope (Zeiss, Oberkochen, Germany) fitted with a 40 × water-immersion objective, differential

interference contrast INCB024360 mouse (DIC) and an infrared filter. The image of the microscope was enhanced using a QICAM camera (QIMAGING, Surrey, BC, Canada) and was displayed with Qcapture Pro 6 on a computer. Pipettes were pulled on a P-87 micropipette puller (Sutter Instruments, Novato, CA, USA) using borosilicate glass capillary

tubing (2.0 mm OD, 1.16 mm ID; Hilgenberg, Malsfeld, Germany). The resistance of the electrodes was 2–5 MΩ when filled with the intracellular solution: (in mm) KMeSO4, 135; KCl, 10; HEPES, 2; MgCl2, 2; ATP-K2, 2; GTP-Na, 0.4; EGTA, 0.1; and biocytin, 0.1% (pH 7.4). Intracellular pipette solutions with low calcium-buffering capacity (0.1 mm EGTA) were used in order to avoid non-physiological calcium buffering (Wolfart et al., 2001). The recordings were confined aminophylline to the ventromedial subdivision of the DRN, which contains the densest cluster of 5-HT neurons (Crawford et al., 2010). A visualized cell was approached with the electrode, a gigaohm seal was established, and the cell membrane was ruptured to obtain the whole-cell configuration. Membrane potentials and currents were recorded using an EPC9 amplifier (HEKA, Lambrecht/Pfalz, Germany) connected to Patchmaster software (HEKA). Liquid junction potentials were corrected. Once the whole-cell recording was obtained, cell characteristics were recorded in current-clamp before adding drugs and either pursuing in current clamp or switching to voltage clamp. Only recordings in which the series resistance was < 30 MΩ and remained stable over time (variations ≤ 20%) were used.