5.6.3. ART should be continued in all women who commenced HAART for PMTCT with a CD4 cell count of between 350 and 500 cells/μL during pregnancy who are coinfected with HBV or HCV in accordance with the MS-275 supplier BHIVA guidelines for the treatment of HIV-1 positive adults with antiretroviral therapy 2012 ( www.bhiva.org/PublishedandApproved.aspx ). Grading: 1B There is evidence that continuing ART in patients coinfected with HBV or HCV reduces co-morbidity progression. For HBV, there is

the additional requirement of viral suppression from antiviral drugs (emtricitabine, lamivudine, tenofovir) and the risk of a flare of hepatitis if discontinued (see Section 6.2 Hepatitis C). 5.6.4 ART can be continued in

all women who commenced HAART for PMTCT with a CD4 cell count of between 350 and 500 cells/μL during pregnancy. Grading: 2C On the basis of the above cohort data the Department of Health and Social Services (2011) [153] and International AIDS Society (2010) guidelines [154] for treating adults have now altered their recommendation and advise treating all adults with a CD4 cell count <500 cells/μL. Moreover, two recent retrospective reviews in women discontinuing ART postpartum found an increased risk of death or opportunistic this website infection among women stopping therapy after delivery. The Tennessee study reviewed patients who discontinued therapy postpartum (mean nadir CD4 cell count 332 cells/μL) in an observational cohort of mothers from 1997 to 2008 [145]. Despite being a small cohort (n = 123), the findings indicated an increased rate of AIDS-defining events and death, and non-AIDS-defining events and death, were more frequent in those discontinuing (n = 54) than in those continuing (n = 69), although this was not statistically significant. This is the only study that has examined the use of HAART on clinical outcomes in women with high CD4 cell counts. However, there were many potential

confounders. In a further retrospective study on mothers discontinuing therapy between 1997 and 2005 GPX6 [147], more opportunistic infections and deaths were found in those who discontinued; however, this was a small, uncontrolled review where 46% had previous ART exposure and 36% a pre-ART CD4 cell count of <350 cells/μL. Lastly, in a large cohort of women who were enrolled in South America and followed up for 6–12 weeks after discontinuation of ART given to prevent MTCT, significant falls in the CD4 cell percentage were seen as would be expected [146]. Other studies have shown no detrimental effects on disease progression in discontinuing treatment postnatally.

Moreover, Crenolanib datasheet studies in animals demonstrated that the BLA is particularly critical for normal performance in a variety of settings that require knowledge of current outcome values including reversal learning and second-order conditioning (Lindgren et al., 2003; Schoenbaum et al., 2003; Johnson et al., 2009). Thus, our finding of a predictiveness signal in the BLA supports the view that the predictive value of CSs is critical for amygdala responses during fear conditioning. On the one hand, the BLA has been highlighted as a site of plasticity in associative learning that is relevant for learning and maintaining CS–US associations (Maren

& Quirk, 2004; Reijmers et al., 2007; Ehrlich et al., 2009; Pape & Pare, 2010), and CS and US information is assumed to converge in this region (Barot et al., 2008). Thus, increasing predictiveness and concomitant increased BOLD responses in the BLA might reflect strengthening of the associative memory with regard to CS–US contingencies. This assumption would, however, require that associative learning also http://www.selleckchem.com/products/bgj398-nvp-bgj398.html occurs in the CS– condition as the predictiveness signal shows equal characteristics for CS100 and CS–. On the other hand,

some recent studies demonstrated that learning of CS–US associations increased over time, when subjects were contingency aware (Schiller et al., 2010; Raio et al., 2012). These findings reflect the observed time course of the predictiveness signal in the current study. Predictiveness might therefore also reflect contingency awareness, which is likely to increase with increasing reliability of outcome predictions. To strengthen the finding of separate recruitment of the BLA and CM by predictiveness and surprise signals, we directly compared the mean activity in both regions. Unsigned PEs were found to correlate with signal changes in the CM but not BLA, whereas the opposite was true for predictiveness signals indicating a clear functional dissociation of both regions. With respect to interactions between the BLA and CM during the process of aversive learning in humans, we can only speculate

as the current study does not allow the drawing of firm conclusions. However, as projections from the Diflunisal BLA to the CM are not reciprocated in the amygdala (Pape & Pare, 2010), we would assume that the surprise signals in the CM project onto cortical areas, which then project back to the BLA where predictiveness as a derivative of these signals controls learning of cue–outcome associations. To summarize, we extended recent findings of PH-like learning signals in the amygdala (Li et al., 2011) by investigating CS- and US-related processing in an RW/PH hybrid model of reinforcement learning. By combining this approach with high-resolution fMRI, we demonstrate a unique functional dissociation of amygdala subregions during associative learning in humans.

, 2005) and (4) diverse adhesive factors (Cegelski

et al., 2008) against various human pathogens. Pseudomonas aeruginosa is an opportunistic human pathogen that readily develops antibiotic resistance and it is a lethal pathogen of particular importance in cystic fibrosis patients (Stover et al., 2000). The bacterium produces a variety of virulence factors, such as Pseudomonas quinolone signal (PQS) (Mashburn & Whiteley, 2005), pyocyanin (Hassett et al., 1992), rhamnolipids (Zulianello et al., 2006), elastase (Pearson et al., 1997) and two endogenous siderophores, pyoverdine and pyochelin (Michel et al., 2005), which are involved in chronic infection (Ben Haj Khalifa et al., 2011). Pseudomonas aeruginosa also produces adhesion factors, exotoxin A, phospholipase C for hemolysis, and exoenzyme S, which are involved in acute infection (Ben Haj Khalifa et al., 2011). Furthermore, biofilm cells

are up to 1000 times more PI3K Inhibitor Library solubility dmso resistant to antibiotics than planktonic cells are (Mah & O’Toole, 2001) and biofilm formation plays an important role in pathogenesis (Rasmussen & Givskov, 2006). Previously, several natural compounds have been reported to decrease the virulence and antibiotic-resistant biofilm formation of P. aeruginosa without affecting its growth; for example, natural brominated furanones produced by the red macroalga Delisea pulchra (Hentzer et al., 2003), d-amino acids (Kolodkin-Gal et al., 2010), cis-2-decenoic acid (Davies & Marques, 2009), corosolic acid and selleck products asiatic acid (Garo et al., 2007). Indole is produced by over 85 species of Gram-positive and Gram-negative bacteria with diverse roles, but P. aeruginosa does not synthesize indole (Lee & Lee, 2010). Previously, natural indole and 7-hydroxyindole diminished the virulence of P. aeruginosa by repressing quorum-sensing-related genes and reduced pulmonary colonization of P. aeruginosa in guinea pigs (Lee et al., 2009). However, these indole compounds increased antibiotic resistance and biofilm formation of P. aeruginosa, probably due to its ecological defense in multispecies nature (Lee et al., 2009), which is a defect of indole

as an antivirulence compound. A natural indole also increased the long term population-wide antibiotic resistance in Escherichia coli (Lee et al., Glutathione peroxidase 2010). Although plant auxin 3-indolylacetonitrile decreased biofilm formation and the production of virulence factors, its virulence reduction is far less efficacious than that of indole (Lee et al., 2011). Therefore, the use of natural indole derivatives is limited due to the natural defense systems of P. aeruginosa. The goal of this study was to identify a novel and potent antivirulence compound against the human pathogen P. aeruginosa. Thirty-one natural and synthetic indole derivatives were initially screened for the inhibition of biofilm formation and hemolytic activity of P. aeruginosa.

In audition, the effects of attention have only been shown in humans when the experimental

task requires sound localization. Studies in monkeys with the use of similar cues but without a sound localization requirement have produced negative results. We have studied the effects of predictive acoustic cues on the latency of gaze shifts to visual and auditory targets in monkeys experienced in localizing sound sources in the laboratory with the head unrestrained. Both attention capture and IOR were demonstrated check details with acoustic cues, although with a faster time course than with visual cues. Additionally, the effect was observed across sensory modalities (acoustic cue to visual target), suggesting that the underlying Torin 1 molecular weight neural mechanisms of these effects may be mediated within the superior colliculus, a center where inputs from both vision and audition converge. “
“Persistent spiking activity is

thought to be a cellular process involved in working memory. We have been interested in whether persistent activity also exists in cortical areas which are not involved in this memory process. To study the possible presence and the mechanisms of persistent activity in layer 5 pyramidal cells of the mouse primary somatosensory, visual and motor cortices, we used patch-clamp and calcium imaging techniques. A combination of cholinergic receptor activation and suprathreshold depolarization or sufficient extracellular stimulation leads to either a subthreshold afterdepolarization or suprathreshold persistent activity in these cortices. There is a continuum of response amplitudes depending on depolarization size. To initiate persistent activity, spikes have to Etomidate be induced at a frequency of at least 20 Hz, if tested for 1 s. Acetylcholine muscarinic, but not nicotinic, receptors are important for initiating persistent activity. Persistent activity is an intrinsic cellular, not a network, phenomenon

as it persists under blockade of ionotropic glutamate and GABA receptors. A rise in intracellular calcium concentration through voltage-gated calcium channels is needed for persistent activity initiation, while intracellular calcium stores are not crucial. The increased intracellular calcium concentration leads to the activation of calcium-sensitive nonspecific cationic channels. This study for the first time describes the presence and the underlying mechanisms of persistent activity in pyramidal cells of three primary sensory and motor cortex areas. These results thereby suggest that persistent activity may be a general capability of deep layer cortical pyramidal cells. “
“Principles of brain function can be disclosed by studying their limits during performance. Tactile stimuli with near-threshold intensities have been used to assess features of somatosensory processing.

, 2005; Erb et al., 2007, 2009; Berg & Ivanovsky, 2009; Peyraud et al., 2009; Alber, 2011; Khomyakova et al. 2011). It is interesting that some intermediates of these assimilatory pathways, for example malate and glyoxylate, are also intermediates in the serine cycle and as such may Selleck PD0325901 afford easy coupling with utilization of the serine cycle. Identification of

acetate utilization pathways in methanotrophs, however, has been challenging. For example, early enzymatic work on M. silvestris found no evidence for the key enzymatic activities in the glyoxylate cycle, i.e., isocitrate lyase and malate synthase (Dunfield et al., 2003; Theisen et al., 2005). Genomic analyses, however, show that genes encoding for these enzymes are present (Chen et al., 2010a). Subsequent deletion of the gene encoding for isocitrate lyase severely limited growth of M. silvestris Venetoclax mouse on acetate, and abolished it on methane (Crombie & Murrell, 2011). As discussed by the authors, such data suggest that the glyoxylate shunt may be vital to M. silvestris for regeneration of glyoxylate in the serine cycle used for carbon assimilation from C1 compounds as well as from C2 compounds. These findings also suggest that this microorganism may have multiple mechanisms to utilize multicarbon

compounds, as growth still occurred on acetate when the gene encoding for isocitrate lyase was deleted. However, homologs of known key genes of ethylmalonyl-CoA, citramalate, and methylaspartate pathways for carbon assimilation from acetate are not readily apparent in the genome sequence of M. silvestris. In contrast,

phylogenetically closely related methylotrophs such as the alphaproteobacterium M. extorquens AM1 were often shown to utilize the coupled serine and ethylmalonyl-CoA pathways for growth (Peyraud et al., 2009; Ŝmejkalová et al., 2010). Preliminary analysis of publicly available genome sequences Cyclic nucleotide phosphodiesterase of obligate methanotrophs [i.e. Alphaproteobacteria Methylosinus trichosporium OB3b (Stein et al., 2010), Methylocystis sp. strain ATCC 49242 (Stein et al., 2011), Gammaproteobacteria M. capsulatus Bath (Ward et al., 2004), Methylobacillus flagellatus KT (Chistoserdova et al., 2007), Methylobacter tundripaludum SV96, Methylomicrobium album BG8, Methylomonas methanica MC09, as well as Candidatus Methylomirabilis oxyfera (Ettwig et al., 2010) and Methylacidiphilum infernorum V4 (Hou et al., 2008)], indicates that the key genes of the ethylmalonyl-CoA pathway (Fig. 3) are only present in the two alphaproteobacterial methanotrophs that were sequenced so far, and are found in synteny in the Methylocystis strain. Further, no evidence was observed for the presence of the set of key genes defining citramalate (Fig. 4) or methylaspartate pathways (Fig. 5) for multicarbon assimilation in any methanotroph for which a genome sequence is available. At present, however, such observations should be treated with caution. First, sequence information is still lacking for some reactions (e.g.

Effectiveness of the Semi-Latin square experimental design. Data S2. Effectiveness of TOT and TC manipulations. Table S1. General matrix for the analysis on the effect of the experimental

series. Table S2. Effects of the experimental Selleckchem Nintedanib conditions (p-values) for each for each dependent variable and location in the sequence. Table S3. Saccadic, microsaccadic, and drift parameters. “
“Recent work has shown that infusion of brain-derived neurotrophic factor (BDNF) into the ventral tegmental area (VTA) promotes a switch in the mechanisms mediating morphine motivation, from a dopamine-independent to a dopamine-dependent pathway. Here we showed that a single infusion of intra-VTA BDNF also promoted a switch in the mechanisms mediating ethanol motivation, from a dopamine-dependent to a dopamine-independent pathway (exactly opposite to that seen with morphine). We suggest that intra-VTA BDNF, via its actions on TrkB receptors, precipitates a switch similar to that which occurs naturally when mice transit from a drug-naive, non-deprived state to a drug-deprived state. The opposite switching of the mechanisms underlying morphine and ethanol motivation by BDNF in previously non-deprived animals is consistent with their proposed AZD9291 chemical structure actions on VTA GABAA receptors. “
“Cerebellar Purkinje cells (PCs) are particularly sensitive to cerebral ischemia, and decreased

GABAA receptor function following injury is thought to contribute to PC sensitivity to ischemia-induced excitotoxicity. Here we examined the functional properties of the GABAA receptors that are spared following ischemia in cultured Purkinje cells from rat and in vivo ischemia

in mouse. Using subunit-specific positive modulators of GABAA receptors, we observed that oxygen and glucose deprivation (OGD) and cardiac arrest-induced cerebral ischemia cause a decrease in sensitivity to the β2/3-subunit-preferring compound, etomidate. However, sensitivity to propofol, a β-subunit-acting compound that modulates β1–3-subunits, was not affected by OGD. The α/γ-subunit-acting compounds, diazepam and zolpidem, were also unaffected by OGD. We performed Alanine-glyoxylate transaminase single-cell reverse transcription–polymerase chain reaction on isolated PCs from acutely dissociated cerebellar tissue and observed that PCs expressed the β1-subunit, contrary to previous reports examining GABAA receptor subunit expression in PCs. GABAA receptor β1-subunit protein was also detected in cultured PCs by western blot and by immunohistochemistry in the adult mouse cerebellum and levels remained unaffected by ischemia. High concentrations of loreclezole (30 μm) inhibited PC GABA-mediated currents, as previously demonstrated with β1-subunit-containing GABAA receptors expressed in heterologous systems.

These include filtration methods

(Hahn et al., 2004), density-gradient centrifugation or elutriation and extinction-dilution whereby samples are diluted, ideally down to single cells, before their culture in isolation (Watve et al., 2000; Connon & Giovannoni, 2002; Ben-Dov et al., 2009; Song et al., 2009; Wang et al., 2009). Many bacteria, particularly those that are oligotrophic in the environment, are very slow-growing. Extended incubation times are a prerequisite 5-Fluoracil mw for the cultivation of such bacteria, with the added benefit that faster-growing members within the mixed populations progressively die off over time, reducing the bacterial competition. The culture of soil bacteria for up to 12 weeks has revealed increasing colony counts and an increased recovery of rarely isolated strains with Selleck Navitoclax time (Davis et al., 2005). Similarly, long-term incubation for up to 24 weeks has been successful for the isolation of strains from the SAR11 clade (Song et al., 2009). Even members of the TM7 Division, which have yet to be cultivated in isolation, were

able to form colonies visible to the naked eye when incubation times of 50 days were used [unpublished observation reported in a review by Hugenholtz (2002)]. Many bacteria have specific nutrient or chemical requirements for growth (Graber & Breznak, 2005; Tripp et al., 2008). For example, members of the genera Abiotrophia and Granulicatella, previously known as the nutritionally variant streptococci, require pyridoxal or l-cysteine for growth (Ruoff, 1991), while Tannerella forsythia requires an exogenous source of N-acetyl muramic acid (Wyss, 1989). The characterization of phylogenetically related species may provide clues to the metabolic requirements of organisms that are so far resistant to culture. Cultivation

media may be modified or enriched with this in mind, resulting in the isolation of previously ‘unculturable’ organisms Ribonucleotide reductase (Sait et al., 2002; Davis et al., 2005). However, simply adding the required substrate to cultivation media may not, in all cases, enable culture of the target organism. For example, slow-growing acetotrophs of the genus Methanosaeta are often outcompeted by faster-growing Methanosarcina spp. in mixed culture. On the other hand, Janssen (2003) found that the incorporation of acetone and isopropanol as enrichments led to the production (by species that ferment these substrates) of a slow and steady source of acetate that allowed Methanosaeta spp. to flourish. Because of a reliance on beneficial bacterial interactions within the source environment, attempts to cultivate certain bacteria under laboratory conditions have sometimes been met with success only when these bacteria are cocultivated with helper strains (Ohno et al., 1999, 2000; Nichols et al., 2008).

In brief, Asp-535, His-538, Glu-542, His-551, Lys-564 and Arg-570 were altered to alanine with both strands harbouring a mutation in the middle were synthesized and used in PCR. Construct pJSR3 (for endogenous toxic studies) and pJC4 (for protein purification) were used as template to amplify a double-stranded nicked circle using different primers as listed in Table 1 resulting in pD535A, pH538A, pE542A, pH551A, pK564A, pR570A and pJC4(D535A), pJC4(H538A), pJC4(E542A), ZD1839 research buy pJC4(H551A), pJC4(K564A), pJC4(R570A), respectively. All the constructs of pBAD were transformed

in E. coli TOP10 resulting in D535A, H538A, E542A, H551A, K564A and R570A strains, and constructs in pET28 were transformed in BL

21 (DE3) pLysS resulting in JC4(D535A), JC4(H538A), JC4(E542A), JC4(H551A), JC4(K564A) and JC4(R570A) strains. All the strains and plasmid used in this study are listed in Table 2. Endogenous toxicity assays were performed Maraviroc chemical structure in E. coli TOP10, as all the constructs of pBAD were transformed in E. coli TOP10 resulting in D535A, H538A, E542A, H551A, K564A and R570A strains. For endogenous toxicity assay, overnight cultures were diluted 100-fold in fresh medium and grown till log phase [optical density at 600 nm (OD600) = 0.4–0.5] and then diluted again to OD600 = 0.01 in fresh medium with 0.2% l-(+)-arabinose (Sigma, St. Louis, MO). Optical density was monitored at 600 nm using a spectrophotometer.

All cultures were grown at 37 °C in LB medium containing 100 mg of ampicillin mL−1, with continuous shaking of ≥ 225 r.p.m. All the experiments were performed in triplicates, and mean values of three results were used to show the growth in percentage (%) at different interval of time. In vitro RNase degradation assay was performed as per protocol described earlier (Singh & Banerjee, 2008) with purified recombinant wild-type catalytic domain and its all mutant variants. Briefly, RNase activity was measured using total MG 132 bacterial RNA from E. coli strain BL 21(DE3)/pLysS as the substrate. The reaction mixture (20 μL) contained 1.2 μg of RNA in 50 mM Tris–HCl buffer (pH 7.5), 50 mM NaCl, 5 mM EDTA and the protein sample to be tested. After 1.5 h of incubation at 37 °C, 2.5 μL of the loading buffer (40% sucrose, 0.125 M EDTA, 0.5% sodium dodecyl sulfate; pH 8) was added, and the mixture was heated at 95 °C for 2 min and resolved on a 1% agarose gel containing ethidium bromide. Intrinsic tryptophan fluorescence spectra of wild-type catalytic domain and its mutants were measured by Varian spectrofluorometer. Spectra were recorded in 20 mM sodium phosphate buffer at protein concentration of 1 μM using excitation wavelength of 295 nm with excitation and emission slit width set at 5 nm.

Co-injection of the same serotype resulted in a high degree of co-infection. Conversely, Osimertinib mouse different serotypes transduced largely non-overlapping populations. These natural preferences

offer the possibility of expressing different transgenes presynaptically and postsynaptically. Luo and colleagues achieved a similar outcome with a Cre-based system that randomly excised one of two stop cassettes to differentially label neurons with red or yellow fluorescence (Zong et al., 2005). Our system now allows this dual mosaic labeling in wild-type mice (or used in addition to germline manipulations), and offers the flexibility to independently control the density of both labels. The ability to express polycistronic transcripts from a single viral promoter also makes it easy to design AAV vectors that both genetically modify and fluorescently label the transduced cells,

as we show through the reliable co-expression of tTA or Cre with YFP or tdTomato. This method allows genetically manipulated and wild-type cells to be distinguished for morphological analysis, electrophysiological studies, or even fluorescence-activated cell sorting (Lobo, 2009; Yang et al., 2011). Although AAV has a relatively small packaging limit compared with other viral vectors (Natkunarajah et al., 2008; Karra & Dahm, 2010), the construct we used allowed 2.3 kb of cDNA to be inserted in addition to the 716 bp YFP coding sequence, 937 bp chick β-actin promoter and 600 bp post-transcriptional regulatory element (woodchuck hepatitis post-transcriptional regulatory element). In theory, Glutamate dehydrogenase proteins up to 800 amino acids long could be incorporated into the construct and still MG-132 supplier allow fluorescent labeling of transgenic cells. Smaller promoters like synapsin-1 would further increase capacity (Shevtsova et al., 2005). In addition to the size barrier, another perceived disadvantage of AAV particularly for developmental studies was the reported

delay between injection and expression. Past work suggested that AAV-encoded fluorescent proteins could take up to 1 week to appear, with peak expression several weeks after onset (Sarra et al., 2002; McCarty et al., 2003; Natkunarajah et al., 2008). In contrast, we show that functional Cre recombinase was present within 2 days of injection, and by P7 the distribution of fluorescent reporter was similar to the adult. This timing is better aligned with the 24 h onset reported by Pilpel et al. (2009) following neonatal injection of AAV8 encoding a fluorescent label under the control of the human synapsin promoter. Although in-utero injections are still needed to manipulate embryonic development, the rapid onset of AAV expression makes neonatal injection an attractive alternative for postnatal studies. Finally, we demonstrate that neonatal injection can be used to label neurons sparsely and brightly enough for in vivo two-photon imaging of neuronal morphology.

When inoculated individually,

nodulation of each mutant was similar to the parental strains. To evaluate competition for nodulation, we inoculated soybean plants with mixtures containing each parental strain together with each derived mutant, and identified the bacterial strains occupying each nodule by their antibiotic resistances. In these experiments, an PD-0332991 cell line Sm-resistant parental strain competed against mutant derivatives that were also resistant to Sm plus another antibiotic (Table 1). Therefore, the antibiotic resistances observed from a nodule where both competitor strains were present simultaneously (double occupation) are the same as from a nodule occupied solely by the mutant. To take into consideration the proportion of nodules with double occupation, we took into account our previous experience with different strains, where we observed an average ± [2 × SEM] of 15.1 ± 4.4% double occupation (Lodeiro et al., 2000b; López-García PCI-32765 mouse et al., 2001, 2002). Thus, to avoid underestimation of wild-type competitiveness, we took the upper limit and assumed 20% double occupation for the χ2 analysis. Hence, we postulated as null hypothesis that 60% of nodules contained bacteria expressing the antibiotic markers of the mutant and the wild type, and the remaining 40% contained rhizobia that express only the

wild-type marker. The results are shown in Table 2. When vermiculite was at field capacity, each flagellin made a different contribution to competitiveness. The strains LP 6865 and LP 6866, which expressed only the thick flagellum, being less motile than their parental strains,

were more competitive for nodulation, while mutants LP 5843 and LP 5844, which expressed only the thin flagellum, were less competitive than the parental strains. Surprisingly, mutants LP 6543 and LP 6644, devoid of both flagella, occupied around 50% of the nodules. Differences of clonidine statistical significance among competitions of double mutants against LP 3004 or LP 3008 might reflect that both the χ2 values calculated were close to the threshold of significance for the tabulated χ2 value. Nevertheless, the trend was clear in that none of the nonmotile double mutants was completely displaced by the wild-type parental strain. To investigate whether this high competitiveness of nonmotile mutants was related to the water contents of pots, we co-inoculated LP 3004 and LP 6543 (nonmotile, lacking both flagella) in vermiculite pots maintained in one of three watering regimens: regularly watered, watered with a double frequency, and flooded. Between days 3 and 12 after inoculation, which is the period where initial nodulation occurs, there was a significant difference in the water status between pots irrigated normally and pots irrigated with double frequency (Fig. S3). In regularly watered pots, the nodule occupation by the nonmotile mutant (plus double occupation) was 53.