The results

were evaluated with a cut-off titer of 1:256,

The results

were evaluated with a cut-off titer of 1:256, as recommended by the manufacturer. Titers between ≥1:16 and ≤1:128 were considered borderline positive. All US evaluations were performed by radiologists. PAIR was conducted under US guidance when there were no contraindications (eg, communication with the bile ducts). The procedure was performed at the Department selleck of Ultrasound, Rigshospitalet, without sedation of the patient and without assistance from anesthesiology staff in the examination room (although assistance was readily available should it be required). Informed consent was obtained from the patient. Intravenous access to a peripheral vein was established and adrenalin 1 mg/mL for intravenous administration was available in case of an anaphylactic reaction. The area of skin chosen for puncture was tagged and disinfected with a 70% ethanol solution. After injection of local anesthetic (10 mL of lidocaine

10 mg/mL), the cyst was punctured with a five to six French see more pig-tail catheter under US guidance. As much cyst material as possible was aspirated, inspected for bilirubin, and collected for subsequent microscopy for the presence of free “hooks” from scolices or scolices themselves. Hypertonic saline (20%) in an amount equalling half the amount of aspirated cystic fluid was injected into the cyst cavity, where it remained for 25 to 30 minutes before being re-aspirated. The catheter was removed and the liver reexamined by US for acute bleeding. The patient rested in bed for 4 h following the procedure. The cyst material was collected for histological and chemical

analysis at the Department of Pathology, Rigshospitalet. The criterion for cure after PAIR was permanent solidification of the cyst(s) (stage CE4/CE5). Before 2002, CE was primarily treated with surgery in our center. From 2002 and onwards, PAIR was chosen as a primary treatment Gefitinib order whenever possible. Surgical treatment was chosen if the cyst communicated with the biliary system or was inaccessible to PAIR due to lack of a viable access for anatomical reasons. Surgical procedures were decompression of the cyst with instillation of 10% saline; removal of cyst contents followed by marsupialization and omentoplasty; or radical liver resection.4 For surgery, criteria for cure were disappearance or solidification of the original cyst cavity. Descriptive statistics were calculated using Microsoft Excel 2000 (Redmond, WA, USA). Fisher’s exact test was applied to compare proportions. Most (22/26) patients had only one cyst, three had two cysts, and one had three cysts. Ten patients were male and 16 were female. Median age at the first presentation of the disease was 36 years (interquartile range 29–45 y). Exposure to risk factors included living in close contact with sheep and dogs.

All standard methods used were performed according to the establi

All standard methods used were performed according to the established protocols (Sambrook et al., 1989). Following the shotgun sequencing of A. halophytica, an open reading frame of 1284 base pairs encoding 427 amino acids of ApSHMT was identified (accession number, AB695121). Amino acid sequence of ApSHMT showed 81% identity with other cyanobacterial SHMTs, such as the Synechococcus sp. PCC 7002. The identity was decreased to 59, 57, 56, and 42–46% for the SHMT from Bacillus stearothermophilus, E. coli, Burkholderia, and plants, respectively (data not shown). However, the amino acid residues important for the structure and function of SHMT (Y56, D202, and K231 for the interaction with PLP; R64 and D73, inter-subunit interaction; H127, cofactor binding; P258 and R363, substrate interaction; numbering was based on ApSHMT, accession number, AB695121) were highly conserved. Many physiological roles of SHMT have been

reported to date (Wilson et al., 1993; Voll et al., 2005; Ibrutinib ic50 Anderson & Stover, 2009; Bauwe et al., 2010; Beaudin et al., 2011). However, the role of SHMT in salinity stress has not been examined although salt-induced increase in SHMT in Anabaena cells has been reported (Srivastava et al., 2011). Therefore, we first studied the expression dynamics of ApSHMT gene under high salinity condition. The expression of ApSHMT was monitored by RT-PCR using the total RNA extracted from NaCl treated up- and down-shocked cells. As a control, the RNase P gene, AprnpB, was used. The NaCl up-shock caused a rapid induction in the ApSHMT transcript expression within 1 h, continued until 12 h, and slightly decreased at 48 h (Fig. 1a). By contrast, there

was no obvious change in ApSHMT transcripts under NaCl down-shock conditions (data Vasopressin Receptor not shown). We examined in vivo the ApSHMT activity under NaCl up-shock conditions. The ApSHMT activity in A. halophytica cells increased approximately twofold by increasing salinity from 0.5 M NaCl to 2.5 M NaCl (Fig. 1b). To characterize the enzymatic properties of ApSHMT protein, we expressed recombinant ApSHMT with 6×His tag at N-terminus under the control of the cold-inducible promoter in E. coli. The expression of ApSHMT was optimum when 0.1 mM isopropyl thio-β-d-galactoside (IPTG) was added at OD620 nm c. 1.0 and the culture was maintained at 16 °C for 16 h. A protein band with expected molecular mass of 44 kDa was detected on SDS-PAGE (see lane 2 in Fig. 2a). Recombinant ApSHMT protein was purified to homogeneity in a single step from crude E. coli lysate using Ni2+-chelating sepharose chromatography (lane 3 in Fig. 2a). The activity of recombinant ApSHMT was assayed with dl-threo-3-phenylserine or l-serine. The former substrate has been used to investigate the aldolase reaction in bacteria (Misono et al., 2005). The enzyme reaction followed the Michaelis–Menten kinetics.

The objective of the SIMPATAZ study was to determine the effectiv

The objective of the SIMPATAZ study was to determine the effectiveness and safety of ATV-containing regimens in patients whose physician has recommended simplification of their ARV treatment to improve ease of administration, patient satisfaction, tolerability, or lipid profile, while maintaining Selleckchem RAD001 virological suppression. SIMPATAZ was a multicentre, prospective, noninterventional, post-authorization, investigator-sponsored study that enrolled patients taking stable PI-based treatment whose physician recommended simplification of their ARV drug regimen to a boosted

ATV-containing regimen (ATV 300 mg/ritonavir 100 mg once daily). Recruitment started in July 2005 and finished in October 2006. The study was conducted at 32 sites throughout Spain, and the protocol

was approved by the Spanish Agency for Medicines and Healthcare Products and by the ethics committees at the participating sites. Patients were followed up every 4 months for 1 year. At each visit, patients underwent a routine physical examination and data were collected on HIV RNA level, CD4 cell MG-132 count, liver function, glucose levels, lipid values [total cholesterol, low-density lipoprotein (LDL) cholesterol, high-density lipoprotein (HDL) cholesterol and triglycerides], adverse reactions, adherence and satisfaction. Adherence was measured using a validated simplified medication adherence questionnaire (SMAQ) [20], with six qualitative questions on adherence and pills missed during the last week and past 3 months. Satisfaction with ARV treatment was evaluated using an ad hoc questionnaire with six items on a visual scale (1=not satisfied to 5=very

satisfied) for different treatment-related aspects such as ease of administration, tolerability, and disease control as perceived by the patient. Eligible patients were HIV-1-infected adults who had been on their current PI-based regimen (unchanged) for at least 6 months and who had an HIV RNA level below the limit of quantification (LOQ) for at least 4 months before simplification. The decision to switch to an ATV-containing regimen was made before inclusion, and each participant provided signed informed consent. Amino acid Patients were excluded if they were pregnant, had not taken ARV drugs before the study or had previously taken ARV drugs not boosted with ritonavir, or if their life expectancy was<12 months. Other exclusion criteria were noncontrolled diabetes mellitus, current alcohol or drug abuse, acute hepatitis at the beginning of the study or advanced liver disease, specified heart conduction system abnormalities, triglycerides ≥1250 mg/dL, serum creatinine higher than twice the upper limit of normal, aminotransferase levels higher than five times the upper limit of normal, and serum bilirubin levels higher than 3 times the upper limit of normal.

We found that the preferred spatial and temporal frequencies, spa

We found that the preferred spatial and temporal frequencies, spatial resolution and high temporal frequency cutoff of area MT neurons were reduced in aged monkeys, and were accompanied by the broadened tuning width of spatial frequency, elevated spontaneous activity, and decreased

signal-to-noise ratio. These results showed that, for neurons in area MT, aging significantly changed both the spatial and temporal frequency response tuning properties. Such evidence provides new insight into the changes occurring at the electrophysiological level that may be related to the aging-related visual deficits, especially in processing spatial and temporal information. buy Everolimus
“During neuronal maturation, the neuron-specific K–Cl co-transporter KCC2 lowers the intracellular chloride and thereby renders GABAergic transmission hyperpolarizing. Independently of its role as a co-transporter, KCC2 plays a crucial role in the maturation of dendritic spines, most probably via an interaction with the cytoskeleton-associated protein 4.1N. In this study, we show that neural-specific overexpression of KCC2 impairs the development of the neural tube- and neural crest-related structures in mouse embryos. At early

stages (E9.5–11.5), the transgenic embryos had a thinner neural tube and abnormal body curvature. They displayed a reduced neuronal differentiation and altered neural crest cell pattern. At later stages (E11.5–15.5), the transgenic embryos had smaller brain structures and a distinctive cleft

palate. Similar results were obtained using overexpression of a transport-inactive N-terminal-deleted variant of KCC2, implying that the effects were not dependent on KCC2′s role as a K–Cl co-transporter. Interestingly, the neural tube of transgenic embryos had an aberrant cytoplasmic distribution of 4.1N and actin. This was corroborated in a neural stem cell line with ectopic expression of KCC2. Embryo phenotype and cell morphology were unaffected by a mutated variant of KCC2 which is unable to bind 4.1N. These results point to a role of KCC2 in neuronal differentiation Phosphoprotein phosphatase and migration during early development mediated by its direct structural interactions with the neuronal cytoskeleton. KCC2 is a neuron-specific isoform of the K–Cl co-transporters. Its developmental upregulation is temporally associated with maturation of postsynaptic GABAergic inhibition in central neurons (Rivera et al., 1999; reviewed in Blaesse et al., 2009). Functional expression of KCC2 during neuronal development leads to a decrease in the intraneuronal Cl− concentration and, consequently, to a hyperpolarizing shift in the reversal potential of GABAA receptor-mediated currents (EGABA) from depolarizing values that are characteristic for immature neurons.

Previous human brain imaging studies have revealed multiple corti

Previous human brain imaging studies have revealed multiple cortical and subcortical areas that are activated when decision uncertainty is linked to outcome probability. However, the neural mechanisms of uncertainty modulation in different perceptual decision tasks have not been systematically investigated. Uncertainty of perceptual decision can

originate either from highly INK 128 in vivo similar object categories (e.g. tasks based on criterion comparison) or from noise being added to visual stimuli (e.g. tasks based on signal detection). In this study, we used functional magnetic resonance imaging (fMRI) to investigate the neural mechanisms of task-dependent modulation of uncertainty in the human brain during perceptual judgements.

We observed correlations between uncertainty levels and fMRI activity in a network of areas responsible for performance monitoring and sensory evidence comparison in both tasks. These areas are associated with late stages of perceptual decision, and include the posterior medial frontal cortex, dorsal lateral prefrontal cortex, and intraparietal sulcus. When the modulation of uncertainty on the two tasks was compared, dissociable cortical networks were identified. Uncertainty in the criterion comparison task modulated activity in the left lateral prefrontal cortex Phosphoprotein phosphatase related to rule retrieval.

In the signal detection task, uncertainty modulated activity in higher learn more visual processing areas thought to be sensory information ‘accumulators’ that are active during early stages of perceptual decision. These findings offer insights into the mechanism of information processing during perceptual decision-making. “
“Specific motor symptoms of Parkinson’s disease (PD) can be treated effectively with direct electrical stimulation of deep nuclei in the brain. However, this is an invasive procedure, and the fraction of eligible patients is rather low according to currently used criteria. Spinal cord stimulation (SCS), a minimally invasive method, has more recently been proposed as a therapeutic approach to alleviate PD akinesia, in light of its proven ability to rescue locomotion in rodent models of PD. The mechanisms accounting for this effect are unknown but, from accumulated experience with the use of SCS in the management of chronic pain, it is known that the pathways most probably activated by SCS are the superficial fibers of the dorsal columns. We suggest that the prokinetic effect of SCS results from direct activation of ascending pathways reaching thalamic nuclei and the cerebral cortex. The afferent stimulation may, in addition, activate brainstem nuclei, contributing to the initiation of locomotion.

Half a decade has

gone by since the publication of the ge

Half a decade has

gone by since the publication of the genome sequence of Xac (da Silva et al., 2002), and apparently, Selleck Midostaurin its large-scale proteome analyses are confined to a few reports using techniques such as two-dimensional protein gels, yeast two-hybrid, and nuclear magnetic resonance scans for folded proteins (Mehta & Rosato, 2001, 2003; Galvao-Botton et al., 2003; Alegria et al., 2004, 2005; Khater et al., 2007). Moreover, the two last methods used heterologous expression of proteins in organisms different from the one under investigation. Most of the limitation to explore the biology of Xac lies in a complete lack of protein expression systems adapted to this bacterium. Here, we showed

the construction and test of new protein expression vectors dedicated to Xac, and the subsequent utilization of our system to characterize the hypothetical protein XAC3408. XAC3408 is 30% identical (at the amino acid level) to the B. subtilis cell division protein ZapA, and our subcellular localization experiments using GFP-XAC3408 support the hypothesis that XAC3408 is the Xac orthologue of ZapABsu. MS-275 cost ZapA-like proteins are conserved among bacteria, in which they function by promoting the FtsZ bundling and stabilization of FtsZ polymers (Gueiros-Filho & Losick, 2002; Low et al., 2004; Scheffers, 2008). ZapAXac exhibited a localization pattern similar to that observed for ZapABsu (Gueiros-Filho & Losick, 2002), and the availability of Xac mutants labeled at the septum

provides a new perspective for antimicrobial drug development trials with Xac aimed to disrupt cell division. The vectors described here are Phosphatidylinositol diacylglycerol-lyase integrative and allow the ectopic expression of proteins from the amy locus of Xac. Such a strategy has been used extensively in the Gram-positive rod B. subtilis (Lewis & Marston, 1999; Gueiros-Filho & Losick, 2002), and it is believed to avoid disturbances to genes/chromosomal regions that might produce undesirable effects in cell growth and altered phenotypes. Besides, integration into amy has the advantage of allowing the characterization of essential genes, which may not accommodate changes in their coding sequences. Finally, the disruption of amy produces a bacterial phenotype easily detectable on a plate and allows the distinction of insertions that had occurred in amy from those in the gene being under investigation. In the present work, we showed that the α-amylase gene was not essential for Xac to grow on a plate, neither was it found to play any key role during infection, an outcome somewhat expected since it has been demonstrated that in bacteria α-amylases may be essential for growth on starch, but dispensable for growth on other carbon sources (Worthington et al., 2003).

(2004) found that in school-aged children the reaction time in a

(2004) found that in school-aged children the reaction time in a visual task and the amplitude of a late negativity elicited by concurrently presented task-irrelevant novel sounds correlated positively (i.e. the longer the reaction times, the larger the RON responses), indicating that the late negativities elicited by novel sounds are also related to the amount of behavioural distraction caused by the unexpected sound in children. The current study shows that, in addition to novel-sound-elicited P3a, musical home activities are also associated with the reduction in this index of distractibility. It cannot be conclusively disentangled from correlational data whether

the relation between musical activities and the P3a and LDN/RON responses found in the current study is due to changes directly caused by such activities in the neural mechanism underlying these responses. For instance, children who are (perhaps inherently) more accurate selleck products at detecting acoustic changes may be more predisposed to musical play and with their own behaviour encourage their parents to sing to them. However, regardless of the initial impetus that eventually led to the observed relationships, it stands to reason that functional changes induced by musical activities could anti-PD-1 antibody inhibitor be

a contributing factor. Firstly, although the auditory system remains malleable by experience throughout the life span, converging evidence from research on a variety of topics, such as the development of auditory processing after fitting of a cochlear implant (Eggermont & Ponton, 2003), the neural underpinnings of second language learning (Kuhl, 2004), and the effects of early blindness on cortical reorganization (Kujala et al., 2000), indicate that the auditory

system exhibits a high potential for functional plasticity in childhood. Furthermore, the animal literature indicates that an acoustically enriched environment leads to functional changes in auditory cortical areas especially in young animals (Zhang et al., 2001; Engineer et al., 2004). Recent longitudinal studies have provided convincing evidence that formal musical training can lead to functional and structural changes in the brain in childhood (Hyde et al., 2009; Moreno et al., 2009; Gerry et al., 2012). In addition, studies on the tuning of the auditory system to culturally Tyrosine-protein kinase BLK typical features of speech and music indicate that the auditory system shows long-term changes as a result of informal everyday exposure to sounds (Näätänen, 2001; Hannon & Trainor, 2007; Wong et al., 2011) and, further, that these changes may be specific to the most relevant deviance types/acoustic changes in speech vs. music (Tervaniemi et al., 2006, 2009). Although longitudinal studies are needed to conclusively resolve this issue, it seems reasonable that even informal musical experience in the form of musical play and parental singing might affect the responsiveness of the auditory system to acoustic changes.

Such signaling

Such signaling APO866 datasheet has been the focus of intense study because of its promise as a target for the treatment of infections (analogous to static drugs rather than cidal). Since the introduction of penicillin, we have seen the rapid emergence of drug-resistant pathogens, which occurs at a rate far outstripping the development of new means of treatment. Interfering with extracellular signaling to prevent the release

of virulence factors, the formation of biofilms or the morphological changes associated with pathogenesis is expected to circumvent this. Such treatments neither halt cellular division directly nor are they toxic to the cells, which means the selective pressure to evolve mechanisms of resistance is likely to be substantially reduced. With this reduced selective pressure, fewer resistant mutants may be generated, which could potentially prolong the usage of the therapeutics and increase their overall effectiveness. In addition, targeting small-molecule signaling pathways ensures that treatments will be directed specifically at the pathogenic organism, rather than the entire microbiome. Medical science is increasingly becoming aware of the host of problems caused by host

microbiome disruptions due to antibiotic treatment. The authors appreciate the invitation to submit this review and acknowledge the insightful critiques and comments of the anonymous referees. “
“BmpA is CT99021 concentration an immunodominant protein of Borrelia burgdorferi as well as an arthritogenic factor. Rabbit antirecombinant BmpA (rBmpA) antibodies were raised, characterized by assaying their cross reactivity with rBmpB, rBmpC and rBmpD, and then rendered monospecific by absorption with rBmpB. This monospecific reagent reacted only with rBmpA in dot immunobinding and detected a single 39 kDa, pI 4-Aminobutyrate aminotransferase 5.0, spot on two-dimensional immunoblots. It was used to assess the BmpA cellular location. BmpA was present in both detergent-soluble and -insoluble fractions of Triton X-114 phase-partitioned borrelial cells, suggesting that it was a membrane

lipoprotein. Immunoblots of proteinase K-treated intact and Triton X-100 permeabilized cells showed digestion of BmpA in intact cells, consistent with surface exposure. This exposure was confirmed by dual-label immunofluorescence microscopy of intact and permeabilized borrelial cells. Conservation and surface localization of BmpA in all B. burgdorferi sensu lato genospecies could point to its playing a key role in this organism’s biology and pathobiology. The Borrelia burgdorferi B31 genome contains many genes coding for putative lipoproteins (4.9% of the chromosomal genes and 14.5% of the plasmid genes) (Fraser et al., 1997; Casjens et al., 2000). Lipoproteins are usually considered structural components of the cell, but surface-exposed lipoproteins of B.

They were monitored for drowsiness and asked to keep their eyes o

They were monitored for drowsiness and asked to keep their eyes open during TMS. Relaxation of the measured muscle

was controlled by continuous visual EMG monitoring. All participants DAPT wore earplugs to protect them from possible acoustic trauma (Rossi et al., 2009), and reduce contamination of TMS-evoked potentials by auditory responses to the clicks produced by the discharge of the TMS coil. The optimal scalp location, over left M1, for TMS-induced activation of the right first FDI was determined as the scalp location from which TMS induced MEPs of maximum peak-to-peak amplitude in the target muscle. Once the optimal spot was identified, the neuronavigation system was used to ensure consistent coil placement and orientation at the optimal spot (Fig. 1A). Resting motor threshold (RMT) was defined as the lowest stimulus intensity of the Nexstim stimulator capable of inducing MEPs of ≥ 50 μV peak-to-peak amplitude in at least five out of ten trials. Active motor threshold (AMT) was defined as the lowest stimulus intensity of the MagPro stimulator capable of inducing visible buy Luminespib twitches

in the FDI in half of the trials while the participants maintained a contraction of the FDI at approximately 20% of the maximal voluntary contraction (Rossini et al., 1994; Chen et al., 2008). Continuous TBS was applied with parameters similar to those used by Huang et al. (2005) – three pulses at 50 Hz, with an interval of 200 ms between the last pulse of a triplet and the first pulse of a triplet, for a total of 600 pulses (Fig. 1B). The intensity was fixed at 80% of AMT. Due to limitations in our experimental set-up, the interstimulus interval was 240 ms compared with the interstimulus interval of 200 ms in the original paradigm introduced by Huang et al. (2005). Thus, in our cTBS paradigm, the triplet repetition rate was about 4.17 Hz instead of 5 Hz, both frequencies being included in the theta band. To establish a pre-cTBS measure, two batches of

10–30 MEPs were recorded in response to a single pulse of TMS at an intensity of 120% of RMT. The pulses were delivered randomly with interstimulus intervals between 5 and 8 s. Following cTBS, a single batch of this website MEPs was measured immediately after (T0) and then at 5, 10, 20, 30, 40, 50 and 60 min following cTBS. EEG was recorded simultaneously at all these times. In a sub-group of seven subjects, resting eyes-closed EEG was recorded at the beginning of the session and after cTBS. These post-cTBS resting EEG measures were recorded sequentially after the single-pulse TMS batches at T5, T10, T20, T30 and T40. Thus, the TX resting EEG measures (X referring to the time in min) started approximately between X + 2 and X + 6 min after cTBS and lasted 2–4 min. Motor-evoked potential peak-to-peak amplitude was determined automatically using the Nexstim Neurophysiologic Analysis software, but checked trial-by-trial by visual inspection.

, 2008);

however, no Na+/H+ antiporters have been identif

, 2008);

however, no Na+/H+ antiporters have been identified at the molecular level in the extremophiles colonizing the Dagong Ancient Brine Well. In recent years, metagenomic libraries have been widely used for mining novel genes Aloxistatin or products of pharmacological importance directly from some environments without necessarily cultivating microorganisms first (Cardenas & Tiedje, 2008; Vakhlu et al., 2008). Many novel genes have also been identified with this approach (Cowan et al., 2005; Schmeisser et al., 2007). In this study, we constructed a metagenomic library by directly extracting DNA from the brine in the Dagong Ancient Brine Well. Screening of Na+/H+ antiporters was performed by function complementation of the antiporter-deficient Escherichia coli strain KNabc that lacks three major genes, nhaA, nhaB and chaA, coding Na+/H+ antiporters (Nozaki et al., 1996). After the identification of the Na+/H+ antiporter genes, the structure and function of the protein it encoded were analyzed. This is the first report of the identification of a novel Na+/H+ antiporter gene from a metagenome from a special man-made ancient hypersaline environment. The halophile genomic DNAs were prepared from the brine in the Dagong Ancient Brine Well using methods originally described by Moon with modifications (Moon et al., 2004). Briefly, 100-mL samples were Copanlisib cell line centrifuged at 14 000 g and 4 °C for

10 min, and the slurry was resuspended with 5 mL phosphate-buffered saline (pH 7.5) centrifuged at 5 g for 2 min at room temperature. The dispersion was again centrifuged at 14 000 g and 4 °C for 2 min. The bacterial cell pellets obtained were directly used for extracting environmental DNA using the Ultra-Clean Soil DNA Kit (Mo Bio Laboratories, Solana Beach, CA). Total DNA was subsequently subjected to electrophoresis in 0.8% agarose gels and stored

at −20 °C. An overnight culture of E. coli KNabc was inoculated into 100 mL of a modified Luria–Bertani medium (LBK medium) consisting of 1.0% tryptone, 0.5% yeast extract Idoxuridine and 87 mM KCl, and then grown at 37 °C under aerobic conditions to an OD600 nm of 0.4. Cells were harvested by centrifugation at 4000 g for 10 min at 4 °C and washed three times in 10 mL of ice-cold sterile 10% glycerol solution before electrocompetent preparation (Yang et al., 2006). The halophile genomic DNAs were partially digested with Sau3AI to produce 1.5–6 kbp fragments. These DNA fragments were separated by agarose electrophoresis and ligated into pUC18, which had been digested with BamHI and dephosphorylated with bacterial alkaline phosphatase, using T4 DNA ligase (Mayumi et al., 2008). The ligated recombinant plasmids (20–200 ng) were added to 50 μL of competent cells of E. coli KNabc suspension and mixed thoroughly. Electroporation was carried out at field strength of 16 kV cm−1 in combination with an electric resistance of 300 Ω at 25 mF in a 0.1-cm electroporation cuvette.