In

contrast, the glial scar, evaluated by glial fibrillary acidic protein staining, showed its highest intensity 21 Selleckchem Ku 0059436 days post-injury in both models. The number of apoptotic oligodendrocytes, detected by CC1/caspase-3 co-labeling, was increased in both models in all evaluated regions. Finally, the numbers of OPCs, evaluated with the markers Tcf4 and Olig2, were increased from day 2 (Olig2) or day 7 (Tcf4) post-injury (P ≤ 0.05). Our results indicate that TBI induces oligodendrocyte apoptosis and widespread myelin loss, followed by a concomitant increase in the number of OPCs. Prevention of myelin loss and oligodendrocyte death may represent novel therapeutic targets for TBI. “
“Working memory (WM) performance in humans can be improved by structuring and organizing the material to be remembered. For visual and verbal information, this process of structuring has been associated with the involvement of a prefrontal–parietal network, but for non-verbal auditory material, the brain areas that facilitate WM for structured information have remained elusive. Using functional magnetic resonance imaging, this study compared neural correlates underlying encoding and rehearsal of auditory WM for structured and unstructured material.

Musicians and non-musicians performed a WM task on five-tone sequences that were either tonally structured (with all tones Erastin price belonging to one tonal key) or tonally unstructured (atonal) sequences. Functional differences were observed for musicians (who are experts in the music domain), but not for non-musicians – The right

pars orbitalis was activated more strongly in musicians during the encoding of unstructured (atonal) vs. structured (tonal) sequences. In addition, data for musicians showed that a lateral (pre)frontal–parietal network (including the right premotor cortex, right inferior precentral sulcus and left intraparietal sulcus) was activated during WM rehearsal of structured, as compared with unstructured, sequences. Our findings indicate that this network plays a role in strategy-based WM for non-verbal auditory information, corroborating previous results Carbohydrate showing a similar network for strategy-based WM for visual and verbal information. “
“Parkinson’s disease is most commonly modelled via unilateral infusion of the neurotoxin 6-hydroxydopamine (6-OHDA) in the rat, but recent work has been aimed to translate the reproducibility and reliability of the model to the mouse. Here we present the effects of unilateral 6-OHDA lesions to either the medial forebrain bundle or the substantia nigra (SN) in mice, which were trained on a lateralised choice reaction time (RT) task.

However, different conclusions were reached concerning the ratio of synchronous to asynchronous mTOR inhibitor release (synchronicity ratio) and its dependence on the identity of the postsynaptic target cell. Whereas Daw et al. (2009) and Karson et al. (2009) suggested that the synchronicity ratio is independent of the identity of the postsynaptic target cell, Ali and Todorova report that this ratio is larger for synapses formed between

CCK-interneurons than for synapses between CCK-interneurons and pyramidal neurons. Accordingly, they suggest that factors governing asynchronous GABA release are synapse-specific and determined in part by the postsynaptic target. Alternatively, these divergent results may be explained by differences in experimental conditions (room versus physiological temperature, number of presynaptic action potentials, current-clamp versus voltage-clamp recording, and/or age of the animals) and the methods used to quantify asynchronous release. Despite these differences, all three papers unequivocally demonstrate asynchronous release at interneuron-interneuron synapses. Asynchronous transmitter release and modulation of synaptic transmission by presynaptic CB1 receptors are hallmarks of the function of synapses formed by CCK-interneurons. How are these two properties interrelated? Ali & Todorova (2010) found that the CB1 receptor inverse agonist AM-251 increased the synchronicity ZD1839 purchase ratio, whereas the

endocannabinoid anandamide decreased it. This finding raises the interesting possibility that synchronous and asynchronous release are differentially affected during DSI. Whether other presynaptic receptors on the terminals of CCK-interneurons have similar effects needs to be determined. Furthermore, filipin the computational significance of asynchronous GABA release in principal neuron-interneuron networks remains to be elucidated. Ali & Todorova (2010)

suggest that asynchronous GABA release modulates the time windows of inhibition, thereby controlling spike timing among local circuit interneurons. “
“This revised Figure 2A corrects the time-points listed for the studies by Kippin et al . (2005), Tanaka et al. (2007) and Tropepe et al. (1997) in the published paper of Hamilton et al. (2013). The authors apologize for any inconvenience caused by this error. “
“Brain plasticity is a double-edged sword. It allows for individuals to learn and adapt to their environment, but peculiarities may also alter the brain and contribute to maladaptive outcomes. Here, in the very interesting study conducted by Frey and colleagues, the authors used measures derived from event-related potentials (ERPs) to assess visuo-spatial maps within the visual cortex in youths with autism spectrum disorders (ASD) and controls. Based on the observation that some individuals with ASD tend to not fixate on a target (i.e. they exhibit off-center fixations), Frey and colleagues hypothesized that this fixation pattern would impact the development of the visual cortex.

HIVAN is rare in patients with CD4 cell counts >350 cells/μL or with undetectable HIV RNA levels [146]. Patients presenting with higher levels of proteinuria (urine albumin–creatinine ratio >70 mg/mmol or urine protein–creatinine ratio >100 mg/mmol or urine protein excretion >1 g/24 h) or proteinuria with haematuria (urine albumin–creatinine ratio >30 mg/mmol or urine protein–creatinine ratio >50 mg/mmol) or stage 4–5 CKD should be referred for specialist assessment

and a renal biopsy considered; those found to have HIVAN should start ART immediately, irrespective of CD4 cell count. For CKD other than HIVAN, there is limited information on the natural history per se and on whether ART Panobinostat confers renal benefit. Immunodeficiency is a potent risk factor for CKD [147, 148]. The majority of patients with CKD have (nadir) CD4 cell counts <350 cells/μL

and thus qualify for ART as per current treatment guidelines. There are no data Selleckchem Docetaxel to provide guidance on whether HIV-positive patients with (or at risk of developing) CKD benefit from earlier ART initiation. None the less, HIV replication, immune activation and inflammation may play a role in the pathogenesis of kidney diseases or contribute to kidney disease progression in some patients [149]. For this reason, ART should be considered in those presenting with CKD other than HIVAN. Renal transplantation is

the treatment of choice for those requiring renal replacement SPTLC1 therapy. Patients to be considered for renal transplantation are required to have suppressed HIV RNA levels and to have CD4 cell counts >200 cells/μL [150], and should start ART, irrespective of CD4 cell count. We recommend against the use of ARV drugs that are potentially nephrotoxic in patients with stages 3–5 CKD if acceptable alternative ARV agents are available (GPP). We recommend dose adjustment of renally cleared ARV drugs in patients with reduced renal function (GPP). Number of patients with CKD stages 3–5 on ARVs that are potentially nephrotoxic and a record of the rationale. Record in patient’s notes of calculated dose of renally cleared ARVs in patients with CKD stage 3 or greater. There are no data from clinical RCTs to inform ART decisions in patients with CKD. The risk of CKD is increased with older age, reduced estimated glomerular filtration rate (eGFR), hypertension, diabetes and with cumulative exposure to indinavir, TDF, ATV and, to a lesser extent, LPV [151, 152]. Indinavir use is no longer recommended in view of the high incidence of renal complications: crystalluria and pyuria are reported in 20–67% [153-155] and nephrolithiasis, tubulointerstitial nephritis and gradual loss of renal function in 4–33% of patients [153, 156-159].

, 1985b), a difference of 2 °C is equivalent to a 6.4% difference in the denaturant. This could yield bands up to 2 cm apart in a 35–65% gel, and multiple bands per 16S rRNA gene sequence could, therefore,

be anticipated. This would invariably lead to multiple bands per 16S rRNA gene sequence, and an overestimation of the diversity. More importantly, the same sequence would yield different banding patterns for different primer batches. The effect of GC-clamp sequence and length variation on band position was then studied experimentally. The V3–5 region of three separate bacterial species of bacteria was amplified using the five sets of primers, and the products were resolved by DGGE. Each lane contained more than one band (Fig. 2a). Importantly, the profiles based on primer sets varied among each other PD-166866 (Fig. 2b). This indicated that DGGE profile variation is due to variation between GC-clamp

primers rather Tacrolimus ic50 than template DNA. One 16S rRNA gene sequence can, therefore, yield multiple bands. The number and distance between the bands appears to be influenced by the specific batch of primers. Three of the five primers (N1–N3) used had an identical sequence design, but displayed deviation both in DGGE patterns and in sequence integrity. DNA sequencing of amplicon pools revealed variation in the GC-clamp sequence, leading to a series of otherwise identical products with different %GC and therefore Tm. Amplicons derived using primer G1 displayed a similar range of variation in GC-clamp sequence and resulting %GC. Primer F1 products displayed the greatest degree of GC-clamp variation and %GC. This during may be due to several adjacent guanosine residues in primer F1. Whether these deviations from the intended sequence occur during synthesis

of the oligonucleotide or during the PCR process is unclear from the current results. Truncation of GC-clamp PCR amplicons of partial 16S rRNA genes has been reported previously (Nubel et al., 1996), and could be due to premature elongation termination of PCR. DNA synthesizers reportedly experience difficulty adding multiple adjacent guanosine residues (Sheffield et al., 1989), and producers of oligonucleotides warn customers of potential problems with the integrity of products with GC-rich stretches. Multiple adjacent guanosine residues reportedly can form aberrant structures such as guanine quartets (Poon & Macgregor, 1998) or four-stranded tetraplexes (Poon & Macgregor, 2000). These structures could interfere during both oligonucleotide synthesis and PCR. Products of primers N1–N3 and F1 lead to a lower degree of GC-clamp variation, and contain only one di-guanosine. Yet, these primers also yielded multiple bands in pure-culture DGGE of all three species, indicating a range of Tm within the amplicon pool. In lieu of multiple guanosines, the GC clamps contained multiple cytosine residues, which would generate multiple guanosines in the reverse strand.

“
“Leprosy classically presents with cutaneous and neural involvement. Rheumatological manifestations are frequent, although often under-recognized. At times, CX-5461 these may present to a rheumatology clinic prior to the diagnosis of leprosy. Herein, we present our experience with patients referred with various rheumatological disorders who were subsequently diagnosed as having leprosy. This retrospective study (January 2001–September 2010) was carried out at the Department of Clinical Immunology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, in northern India. Patients who were confirmed as having leprosy were included. Details regarding demographic and clinical

presentations were collected. Forty-four cases (30 male, mean age 40 ± 13.6 years and mean disease duration 18.7 ± 24.3 months) were identified. Musculoskeletal manifestations included arthritis (n = 22), swollen hands and feet syndrome (SHFS) (n = 11), tenosynovitis (n = 9), painful swollen feet (n = 9), arthralgias (n = 7) and vasculitis (n = 1). Distribution of joints mimicked rheumatoid arthritis

(n = 14) and spondyloarthropathy (n = 7). Arthritis and/or tenosynovitis were part of spontaneous onset lepra reaction in 28 cases. Other clinical manifestations were: paresthesias find more (n = 28), erythematous nodules (n = 25) and anesthetic patches (n = 7). Thirty-one patients had thickened nerves (ulnar n = 28, common peroneal n = 21). Eight patients did not have any cutaneous manifestations and had presented with SHFS and arthritis or tenosynovitis. These were labeled as pure neuritic leprosy. Most

of the patients responded to multidrug anti-leprosy therapy and glucocorticoids. Rheumatological presentations of leprosy may mimic RA, spondyloarthropathy or vasculitis. Pure neuritic variety and spontaneous type 2 lepra reaction pose unique diagnostic challenges. Increased awareness may avoid delay in diagnosis. Resminostat “
“To assess patient satisfaction with the rheumatology telemedicine service provided to a rural town in northern Australia. A prospective, questionnaire-based exploratory study of patients seen at the Mount Isa (rural town) rheumatology telemedicine clinics during 2012 was undertaken. Control groups included patients travelling over 3 h to be seen face-to-face in Townsville (tertiary referral centre), and patients seen at the infrequent face-to-face clinic in Mount Isa. A 5-point Likert scale was used to explore themes of communication, confidentiality, physical examination, rapport, medication safety and access. This study evaluated 107 rheumatology outpatients (49 telemedicine, 46 face-to-face Townsville, 12 face-to-face Mount Isa). Patients seen in Mount Isa travelled a median of < 10 km for either the telemedicine or local face-to-face appointments. The patients attending the Townsville face-to-face clinic travelled a median of 354 km.

QSS 2009 contacted or attempted to contact 3,112 households; 1,536 subjects declined participation, 142 households could not be contacted and 129 were otherwise ineligible. Thus, the final sample for QSS 2009 included 1,292 respondents, 860 from Southeast Queensland and 432 from Other Queensland for an overall response rate of 41.5%. The sample was nearly DAPT chemical structure equally divided between males and females (50.2%

vs 49.8%). Younger people (aged 18–34 y) were under-represented in the sample; and older people (aged >55 y) were over-represented in the sample; otherwise, the demographics of the participants reasonably approximated that of the general population.9 Responses to the two questions concerning travel and influenza are shown in Table 1; 688 (53.2%) of respondents indicated some level of concern about Pandemic (H1N1) 2009 when traveling and 458 (35.5%)

indicated they would likely cancel their own commercial air travel if they had a cough and fever that lasted more than one day. When cross-tabulating these responses, people who expressed concern regarding Pandemic (H1N1) 2009 when they traveled were more likely than those without concern to cancel their own commercial air travel if they had a cough and fever lasting more than one day (44.7% vs 27.7%, χ2 = 33.53, p < 0.001). Nonetheless, there were 363 respondents who expressed concern regarding Pandemic (H1N1) 2009, but who would not have cancelled their own commercial air travel if they had symptoms selleck chemicals of a viral respiratory infection. Bivariate associations between demographic variables and both concern about and willingness

to cancel travel are shown in Table 2, and the final multivariate models are shown in Table 3. When controlling for covariance and 3-mercaptopyruvate sulfurtransferase confounding, respondents living outside of metropolitan Southeast Queensland (AOR = 0.589; CI: 0.396–0.874), those with more than 14 years of education (AOR = 0.651; CI: 0.444–0.952), and those with incomes greater than A$100,000 per year (AOR = 0.528; CI: 0.353–0.791) were all less likely to express concern regarding Pandemic (H1N1) 2009 when traveling. There were no interaction effects among these variables. Only age was significantly associated with the likelihood of cancelling travel if a respondent was symptomatic, with younger respondents (18–24 y old) less likely than others to cancel pre-existing travel plans (AOR = 0.469; CI: 0.260–0.847). Previous emerging infectious disease outbreaks, such as severe acute respiratory syndrome (SARS), had far reaching impacts on travel and tourism, particularly, with shutdown of airline travel during the height of the SARS outbreak.10 Avian influenza has not had the same impact; however, it has raised considerable concern among travelers and government travel advisories alike.

6 ± 12.4 years; age range 25–73 years). The duration of the disease ranged from 1 to 30 years. All patients were clinically examined by a rheumatologist who had more than 10 years of relevant clinical experience as a rheumatologist (MS) and was unaware of the US findings. At each clinical examination, 28 joints including the bilateral glenohumeral, elbow, wrist, metacarpophalangeal, proximal interphalangeal joints of the hands, and knee joints, were

assessed for tenderness and swelling. The tender joint count (TJC; range, 0–28) and swollen joint count (SJC; range, 0–28) were recorded for each patient. Each patient provided an overall assessment of their functional status using the global pain intensity visual SCH772984 price analog scale (VAS) score (VAS pain; range, 0–100). The disease activity of each patient was assessed by the Disease Activity Score for 28 joints (DAS28). Tests to determine selleck chemicals the CRP levels and erythrocyte sedimentation rate (ESR) were performed on the same day when both clinical and sonographic examinations were conducted. All subjects were informed of the study procedure and purpose, and written informed consent was obtained from all participants prior to participation. This study was conducted in accordance

with the guidelines of the 1995 Declaration of Helsinki and was approved by the institutional ethics committee. Sonographic examinations were performed using the ProSound Alpha 10 (Hitachi Aloka Medical, Ltd., Tokyo, Japan) with a 6.0–14.0 MHz linear array probe. This examination was performed by a board-certified sonographer (TW) blinded to the clinical information of each patient. Flow-mediated endothelium-dependent vasodilation was measured according to the 2007 Japanese guidelines for US assessment of FMD. FMD HSP90 was measured using brachial US after 15 min of rest in a quiet, dark, temperature-controlled room (25°C). All patients were assessed at similar times of the day. A high-resolution linear array transducer was coupled to computer-assisted

analysis software (e-TRACKING system, Hitachi Aloka Medical, Ltd.) that used an automated edge detection system to measure the brachial artery diameter. Measurements were made from the anterior to posterior interface between the lumen and intima at end-diastole, in synchrony with the electrocardiographic R-wave. The right brachial artery was evaluated with high-resolution US at the elbow, 3–7 cm above the antecubital fossa, where it formed a straight segment in the supine position. The occlusion blood pressure cuff was placed around the right upper forearm, just below the antecubital fossa. The baseline longitudinal image of the artery was acquired for 30 s; the blood pressure cuff was subsequently inflated to 30 mmHg above systolic pressure for 5 min.

Any queries (other than missing material) should be directed to the corresponding author for the article. “
“Helicobacter pylori infects the stomach of about half of the world’s human population, frequently causing chronic inflammation at the origin of several gastric pathologies. One of the most remarkable characteristics of the species is its remarkable genomic plasticity in which homologous recombination (HR) plays

a critical role. Here, we analyzed the role of the H. pylori homologue of the AddAB recombination protein. Bioinformatics analysis of the proteins unveils the similarities and differences of the H. pylori AddAB complex with respect to the Selleckchem Atezolizumab RecBCD and AddAB complexes from Escherichia coli and Bacillus subtilis, respectively. Helicobacter pylori mutants lacking functional addB or/and addA show the same level of sensitivity to DNA-damaging agents such as UV or irradiation and of deficiency in intrachromosomal RecA-dependent HR. Epistasis analyses of both DNA repair and HR phenotypes, using double and triple

recombination mutants, demonstrate that, in H. pylori, AddAB and RecOR complexes define two separate presynaptic pathways with little functional overlap. However, neither of these complexes participates in the RecA-dependent process of transformation of these naturally competent bacteria. The pathogen Helicobacter pylori colonizes the stomach mucosa of about half of the human population, frequently resulting in chronic gastritis, which can lead to peptic ulcers

and, in a small fraction of cases, to cancer. Adaptation of H. pylori to the changing gastric environment within BTK inhibitors library a host, or to new hosts, suggests an enhanced ability of this pathogen to change. Indeed, H. pylori is one of the most genetically diverse bacterial species. At the origin of such diversity are both mutations and recombination events (Suerbaum & Josenhans, 2007). Incorporation of DNA sequences by homologous recombination (HR) into the H. pylori chromosome, facilitated by the natural competence of this species, is crucial for horizontal gene transfer between unrelated strains colonizing the same host (Kersulyte et al., 1999). This process is believed to be the cause of its panmictic population structure (Suerbaum et al., 1998). Analysis of the genomic sequences has also underlined the importance of intragenomic Org 27569 chromosomal rearrangements mediated by HR (Israel et al., 2001; Aras et al., 2003). In Escherichia coli, two major DNA recombination initiation (presynaptic) pathways coexist and are complementary: the RecFOR and the RecBCD pathways. The RecFOR pathway is essential for the postreplication repair of gaps and for the restart of replication following UV damage. However, none of the recF, recO and recR mutants show a decrease in HR following conjugation or transduction (Howard-Flanders & Bardwell, 1981; Kuzminov, 1999; Ivancic-Bace et al., 2003). We recently reported the presence in H.

, 2001a,b; Garcia-Osta et al., Saracatinib 2006; Nikitin, 2007), neuroinflammation

(Cardinaux et al., 2000; Ejarque-Ortiz et al., 2007; Straccia et al., 2011; Fields & Ghorpade, 2012), neurogenesis, and neuronal proliferation and differentiation (Cortés-Canteli et al., 2002; Calella et al., 2007; Aguilar-Morante et al., 2011), whereas its role in neuronal survival/apoptosis remains unclear. In fact, C/EBP β induces the expression of genes involved in brain injury and inflammatory processes; it is upregulated after ischemic injury and in a mouse model of hippocampal kainate excitotoxicity, as well as in adult hippocampal neurogenesis (Cortés-Canteli et al., 2004, 2008, 2011; Sandhir & Berman, 2010; Rininger et al., 2012). In cortical neurons, C/EBP β expression is induced after hypoxic stress, supporting neuronal survival by inhibiting p53 (Halterman et al., 2008). On the other hand, C/EBP β induces apoptosis in neuroblastoma through p53 activation (Cortés-Canteli et al., 2002). In primary cultures of rat cerebellar granule neurons (CGNs), high Ca2+ influx through N-methyl-d-aspartate (NMDA) receptors increases PLX4032 molecular weight nuclear C/EBP β levels and induces excitotoxic neuronal death (Marshall et al., 2003). However, no studies so far have studied the expression of all C/EBP β isoforms in survival/apoptotic conditions. To fill this gap, we used neuronal primary cultures and induced apoptosis, in order

to study the role of C/EBP β isoforms in neuronal survival/death. Primary cultures of CGNs were prepared from 7-day-old Wistar Han Outbred Rat pups derived from a local animal house (Gallo et al., 1987). All animal experiments were authorized

by a local bioethical committee (Protocol no. 17-72-1212), and experiments were carried out in accordance with the European Communities Council Directive of 24 November 1986 (86/609/EEC). Animal health and comfort were veterinarily controlled. For all experiments presented here, a total number of 72 pups were used. Briefly, animals were rapidly anesthetized with an ice-cold treatment, and killed by decapitation; cerebella were removed and dissected from their meninges in Krebs’ buffer containing 0.3% bovine serum albumin (BSA). Cerebella were then dissociated with trypsin at 37 °C for 15 min, and triturated by use of a Pasteur pipette, in a 0.125 mg/mL Resveratrol DNaseI/0.52 mg/mL soybean trypsin inhibitor solution. Dissociated cells were collected by centrifugation, resuspended in Basal Medium Eagle (Invitrogen, DH Breda, NL, USA), supplemented with 2 mm glutamine, 100 μm gentamicin sulfate, 10% inactivated fetal bovine serum (Invitrogen), and 25 mm KCl, and plated in plastic dishes, previously coated with poly-l-lysine (0.01 mg/mL), at a density of 2.2 × 106 cells per 35-mm dish. After incubation for 16 h at 37 °C in a 95% air/5% CO2 (v/v) atmosphere, 10 μm cytosine arabinofuranoside was added to reduce proliferation of non-neuronal cells.

, 2001a,b; Garcia-Osta et al., Bcl-2 pathway 2006; Nikitin, 2007), neuroinflammation

(Cardinaux et al., 2000; Ejarque-Ortiz et al., 2007; Straccia et al., 2011; Fields & Ghorpade, 2012), neurogenesis, and neuronal proliferation and differentiation (Cortés-Canteli et al., 2002; Calella et al., 2007; Aguilar-Morante et al., 2011), whereas its role in neuronal survival/apoptosis remains unclear. In fact, C/EBP β induces the expression of genes involved in brain injury and inflammatory processes; it is upregulated after ischemic injury and in a mouse model of hippocampal kainate excitotoxicity, as well as in adult hippocampal neurogenesis (Cortés-Canteli et al., 2004, 2008, 2011; Sandhir & Berman, 2010; Rininger et al., 2012). In cortical neurons, C/EBP β expression is induced after hypoxic stress, supporting neuronal survival by inhibiting p53 (Halterman et al., 2008). On the other hand, C/EBP β induces apoptosis in neuroblastoma through p53 activation (Cortés-Canteli et al., 2002). In primary cultures of rat cerebellar granule neurons (CGNs), high Ca2+ influx through N-methyl-d-aspartate (NMDA) receptors increases find more nuclear C/EBP β levels and induces excitotoxic neuronal death (Marshall et al., 2003). However, no studies so far have studied the expression of all C/EBP β isoforms in survival/apoptotic conditions. To fill this gap, we used neuronal primary cultures and induced apoptosis, in order

to study the role of C/EBP β isoforms in neuronal survival/death. Primary cultures of CGNs were prepared from 7-day-old Wistar Han Outbred Rat pups derived from a local animal house (Gallo et al., 1987). All animal experiments were authorized

by a local bioethical committee (Protocol no. 17-72-1212), and experiments were carried out in accordance with the European Communities Council Directive of 24 November 1986 (86/609/EEC). Animal health and comfort were veterinarily controlled. For all experiments presented here, a total number of 72 pups were used. Briefly, animals were rapidly anesthetized with an ice-cold treatment, and killed by decapitation; cerebella were removed and dissected from their meninges in Krebs’ buffer containing 0.3% bovine serum albumin (BSA). Cerebella were then dissociated with trypsin at 37 °C for 15 min, and triturated by use of a Pasteur pipette, in a 0.125 mg/mL Ergoloid DNaseI/0.52 mg/mL soybean trypsin inhibitor solution. Dissociated cells were collected by centrifugation, resuspended in Basal Medium Eagle (Invitrogen, DH Breda, NL, USA), supplemented with 2 mm glutamine, 100 μm gentamicin sulfate, 10% inactivated fetal bovine serum (Invitrogen), and 25 mm KCl, and plated in plastic dishes, previously coated with poly-l-lysine (0.01 mg/mL), at a density of 2.2 × 106 cells per 35-mm dish. After incubation for 16 h at 37 °C in a 95% air/5% CO2 (v/v) atmosphere, 10 μm cytosine arabinofuranoside was added to reduce proliferation of non-neuronal cells.