Fitness variation among host species and the paradox of ineffective rhizobia.

Legumes can preferentially select beneficial rhizobial symbionts and sanction ineffective strains that fail to fix nitrogen. Yet paradoxically, rhizobial populations vary from highly beneficial to ineffective in natural and agricultural soils. Classic models of symbiosis focus on the single dimension of symbiont cost-benefit to sympatric hosts, but fail to explain the widespread persistence of ineffective rhizobia. Here, we test a novel framework predicting that spatio-temporal and community dynamics can maintain ineffective strains in rhizobial populations. We used clonal and multistrain inoculations and quantitative culturing to investigate the relative fitness of four focal Bradyrhizobium strains varying from effective to ineffective on Acmispon strigosus. We found that an ineffective Bradyrhizobium strain can be sanctioned by its native A. strigosus host across the host's range, forming fewer and smaller nodules compared to beneficial strains. But the same ineffective Bradyrhizobium strain exhibits a nearly opposite pattern on the broadly sympatric host Acmispon wrangelianus, forming large nodules in both clonal and multistrain inoculations. These data suggest that community-level effects could favour the persistence of ineffective rhizobia and contribute to variation in symbiotic nitrogen fixation.

Epidemic Spread of Symbiotic and Non-Symbiotic Bradyrhizobium Genotypes Across California.

The patterns and drivers of bacterial strain dominance remain poorly understood in natural populations. Here, we cultured 1292 Bradyrhizobium isolates from symbiotic root nodules and the soil root interface of the host plant Acmispon strigosus across a >840-km transect in California. To investigate epidemiology and the potential role of accessory loci as epidemic drivers, isolates were genotyped at two chromosomal loci and were assayed for presence or absence of accessory "symbiosis island" loci that encode capacity to form nodules on hosts. We found that Bradyrhizobium populations were very diverse but dominated by few haplotypes-with a single "epidemic" haplotype constituting nearly 30 % of collected isolates and spreading nearly statewide. In many Bradyrhizobium lineages, we inferred presence and absence of the symbiosis island suggesting recurrent evolutionary gain and or loss of symbiotic capacity. We did not find statistical phylogenetic evidence that the symbiosis island acquisition promotes strain dominance and both symbiotic and non-symbiotic strains exhibited population dominance and spatial spread. Our dataset reveals that a strikingly few Bradyrhizobium genotypes can rapidly spread to dominate a landscape and suggests that these epidemics are not driven by the acquisition of accessory loci as occurs in key human pathogens.

Lotus hosts delimit the mutualism-parasitism continuum of Bradyrhizobium.

Symbioses are modelled as evolutionarily and ecologically variable with fitness outcomes for hosts shifting on a continuum from mutualism to parasitism. In a classic example, rhizobia fix atmospheric nitrogen for legume hosts in exchange for photosynthetic carbon. Rhizobial infection often enhances legume growth, but hosts also incur interaction costs because of root tissues and or metabolites needed to support symbionts in planta. Rhizobia exhibit genetic variation in symbiotic effectiveness, and ecological changes in light or mineral nitrogen availability can also alter the benefits of rhizobial infection for hosts. The net effects of symbiosis thus can range from mutualistic to parasitic in a context-dependent manner. We tested the extent of the mutualism-parasitism continuum in the legume-rhizobium symbiosis and the degree to which host investment can shape its limits. We infected Lotus strigosus with sympatric Bradyrhizobium genotypes that vary in symbiotic effectiveness. Inoculations occurred under different mineral nitrogen and light regimes spanning ecologically relevant ranges. Net growth benefits of Bradyrhizobium infection varied for Lotus and were reduced or eliminated dependent on Bradyrhizobium genotype, mineral nitrogen and light availability. But we did not detect parasitism. Lotus proportionally reduced investment in Bradyrhizobium as net benefit from infection decreased. Lotus control occurred primarily after infection, via fine-scale modulation of nodule growth, as opposed to control over initial nodulation. Our results show how divestment of symbiosis by Lotus can prevent shifts to parasitism.

A new global commitment to disease control in Africa.

Despite the availability of effective therapies in developed nations, infectious diseases continue to take a grave toll on the population and economy of sub-Saharan Africa. Aside from a few successes, the global donor community has not adequately helped African governments meet these health challenges. However, if annual donor contributions increased approximately 10-20-fold, millions of lives could be saved, helping Africa escape the cycle of disease and impoverishment.

Symbiont genomics, our new tangled bank.

Microbial symbionts inhabit the soma and surfaces of most multicellular species and instigate both beneficial and harmful infections. Despite their ubiquity, we are only beginning to resolve major patterns of symbiont ecology and evolution. Here, we summarize the history, current progress, and projected future of the study of microbial symbiont evolution throughout the tree of life. We focus on the recent surge of data that whole-genome sequencing has introduced into the field, in particular the links that are now being made between symbiotic lifestyle and molecular evolution. Post-genomic and systems biology approaches are also emerging as powerful techniques to investigate host-microbe interactions, both at the molecular level of the species interface and at the global scale. In parallel, next-generation sequencing technologies are allowing new questions to be addressed by providing access to population genomic data, as well as the much larger genomes of microbial eukaryotic symbionts and hosts. Throughout we describe the questions that these techniques are tackling and we conclude by listing a series of unanswered questions in microbial symbiosis that can potentially be addressed with the new technologies.

Can Assessment of the Tongue on Brain MRI Aid Differentiation of Seizure from Alternative Causes of Transient Loss of Consciousness?

BACKGROUND AND PURPOSE: Transient loss of consciousness is commonly evaluated in the emergency department. Although typically caused by epileptic seizure, syncope, or psychogenic nonepileptic seizure, the underlying etiology is frequently misdiagnosed. Lateral tongue bites are reportedly a specific clinical finding of seizure. We have observed tongue signal abnormality suggesting bite injury on brain MR imaging after seizures. We hypothesized an association between tongue signal abnormality and seizure diagnosis among patients in the emergency department imaged for transient loss of consciousness. Our purposes were to determine the prevalence of tongue signal abnormality among this population and the predictive performance for seizure diagnosis. MATERIALS AND METHODS: For this retrospective study including 82 brain MR imaging examinations, 2 readers independently assessed tongue signal abnormality on T2-weighted and T2-weighted FLAIR images. Discrepancies were resolved by consensus, and interrater reliability (Cohen κ) was calculated. The final diagnosis was recorded. Proportions were compared using the Fisher exact test. RESULTS: Tongue signal abnormality was present on 19/82 (23%) MR imaging examinations. Interrater reliability was "substantial" (κ = 0.77). Seizure was diagnosed among 18/19 (95%) patients with tongue signal abnormality and 29/63 (46%) patients without it (P < .001). In our cohort, tongue signal abnormality conveyed 97% specificity, 95% positive predictive value, and 63% accuracy for seizure diagnosis. CONCLUSIONS: Tongue signal abnormality was observed in 23% of the study cohort and conveyed 97% specificity and 95% positive predictive value for seizure diagnosis. By assessing and reporting tongue signal abnormality, radiologists may facilitate a timely and accurate diagnosis of seizure among patients imaged for transient loss of consciousness.

Dual-Energy Parathyroid 4D-CT: Improved Discrimination of Parathyroid Lesions from Thyroid Tissue Using Noncontrast 40-keV Virtual Monoenergetic Images.

BACKGROUND AND PURPOSE: In parathyroid CT, a noncontrast phase aids discrimination of parathyroid lesions (not iodine-containing) from thyroid tissue (iodine-containing). When thyroid iodine is pathologically diminished, this differentiation is difficult with standard CT. Because the attenuation of an element is maximal near its K-edge (iodine = 33.2 keV), we hypothesized that dual-energy CT 40-keV virtual monoenergetic images will accentuate thyroid iodine relative to standard images, improving the differentiation of thyroid from parathyroid lesions. Our purpose was to test this hypothesis through quantitative assessment of Hounsfield unit attenuation and contrast-to-noise on dual-energy CT standard (70-keV) and 40-keV noncontrast images. MATERIALS AND METHODS: For this retrospective study including 20 dual-energy parathyroid CTs, we used an ROI-based analysis to assess the attenuation of thyroid tissue, parathyroid lesions, and sternocleidomastoid muscle as well as corresponding contrast-to-noise on standard and 40- keV noncontrast images. Wilcoxon signed rank tests were performed to compare differences between 70 and 40 keV. RESULTS: Absolute and percentage increases in attenuation at 40 keV were significantly greater for thyroid gland than for parathyroid lesions and sternocleidomastoid muscle (P < .001 for all). Significant increases in the contrast-to-noise of thyroid relative to parathyroid lesions (median increase, 0.8; P < .001) and relative to sternocleidomastoid muscle (median increase, 1.3; P < .001) were observed at 40 keV relative to 70 keV. CONCLUSIONS: Forty-kiloelectron volt virtual monoenergetic images facilitate discrimination of parathyroid lesions from thyroid tissue by significantly increasing thyroid attenuation and associated contrast-to-noise. These findings are particularly relevant for parathyroid lesions that exhibit isoattenuation to the thyroid on parathyroid CT arterial and venous phases and could, therefore, be missed without the noncontrast phase.

The Pharyngolaryngeal Venous Plexus: A Potential Pitfall in Surveillance Imaging of the Neck.

BACKGROUND AND PURPOSE: Among patients undergoing serial neck CTs, we have observed variability in the appearance of the pharyngolaryngeal venous plexus, which comprises the postcricoid and posterior pharyngeal venous plexuses. We hypothesize changes in plexus appearance from therapeutic neck irradiation. The purposes of this study are to describe the CT appearance of the pharyngolaryngeal venous plexus among 2 groups undergoing serial neck CTs-patients with radiation therapy-treated laryngeal cancer and patients with medically treated lymphoma-and to assess for changes in plexus appearance attributable to radiation therapy. MATERIALS AND METHODS: For this retrospective study of 98 patients (49 in each group), 448 contrast-enhanced neck CTs (222 laryngeal cancer; 226 lymphoma) were assessed. When visible, the plexus anteroposterior diameter was measured, and morphology was categorized. RESULTS: At least 1 plexus component was identified in 36/49 patients with laryngeal cancer and 37/49 patients with lymphoma. There were no statistically significant differences in plexus visibility between the 2 groups. Median anteroposterior diameter was 2.1 mm for the postcricoid venous plexus and 1.6 mm for the posterior pharyngeal venous plexus. The most common morphology was "bilobed" for the postcricoid venous plexus and "linear" for the posterior pharyngeal venous plexus. The pharyngolaryngeal venous plexus and its components were commonly identifiable only on follow-up imaging. CONCLUSIONS: Head and neck radiologists should be familiar with the typical location and variable appearance of the pharyngolaryngeal plexus components so as not to mistake them for neoplasm. Observed variability in plexus appearance is not attributable to radiation therapy.

Origins of cheating and loss of symbiosis in wild Bradyrhizobium.

Rhizobial bacteria nodulate legume roots and fix nitrogen in exchange for photosynthates. These symbionts are infectiously acquired from the environment and in such cases selection models predict evolutionary spread of uncooperative mutants. Uncooperative rhizobia - including nonfixing and non-nodulating strains - appear common in agriculture, yet their population biology and origins remain unknown in natural soils. Here, a phylogenetically broad sample of 62 wild-collected rhizobial isolates was experimentally inoculated onto Lotus strigosus to assess their nodulation ability and effects on host growth. A cheater strain was discovered that proliferated in host tissue while offering no benefit; its fitness was superior to that of beneficial strains. Phylogenetic reconstruction of Bradyrhizobium rDNA and transmissible symbiosis-island loci suggest that the cheater evolved via symbiotic gene transfer. Many strains were also identified that failed to nodulate L. strigosus, and it appears that nodulation ability on this host has been recurrently lost in the symbiont population. This is the first study to reveal the adaptive nature of rhizobial cheating and to trace the evolutionary origins of uncooperative rhizobial mutants.

Host control over infection and proliferation of a cheater symbiont.

Host control mechanisms are thought to be critical for selecting against cheater mutants in symbiont populations. Here, we provide the first experimental test of a legume host's ability to constrain the infection and proliferation of a native-occurring rhizobial cheater. Lotus strigosus hosts were experimentally inoculated with pairs of Bradyrhizobium strains that naturally vary in symbiotic benefit, including a cheater strain that proliferates in the roots of singly infected hosts, yet provides zero growth benefits. Within co-infected hosts, the cheater exhibited lower infection rates than competing beneficial strains and grew to smaller population sizes within those nodules. In vitro assays revealed that infection-rate differences among competing strains were not caused by variation in rhizobial growth rate or interstrain toxicity. These results can explain how a rapidly growing cheater symbiont--that exhibits a massive fitness advantage in single infections--can be prevented from sweeping through a beneficial population of symbionts.

Internal Auditory Canal Diverticula among Pediatric Patients: Prevalence and Assessment for Hearing Loss and Anatomic Associations.

BACKGROUND AND PURPOSE: Internal auditory canal diverticula are focal lucencies along the anterior-inferior aspect of the internal auditory canal fundus. Studies in adults report conflicting data on the etiology and clinical relevance of this finding. We would expect a pediatric study to help elucidate the significance of internal auditory canal diverticula. The primary goals of this study were to determine the temporal bone CT prevalence of diverticula among pediatric patients and to assess possible hearing loss and anatomic associations. MATERIALS AND METHODS: For this retrospective study including 283 pediatric temporal bone CTs, 4 neuroradiologists independently assessed for diverticula. Discrepancies were resolved by consensus. One neuroradiologist assessed for an enlarged vestibular aqueduct, labyrinthine dysplasia, cochlear cleft, and otospongiosis. Patient demographics, audiologic data, and pertinent clinical history were recorded. One-way analysis of variance and the Fisher exact test were used to assess possible associations between diverticula and specific patient characteristics. RESULTS: Diverticula were observed in 42/283 patients (14.8%) and were more commonly bilateral. There was no significant association with age, sex, hearing loss, enlarged vestibular aqueduct, labyrinthine dysplasia, or cochlear cleft. A statistically significant association was observed with otospongiosis (P = .013), though only 1 study patient had this disease. CONCLUSIONS: Internal auditory canal diverticula are a common finding on pediatric temporal bone CT. In the absence of clinical or imaging evidence for otospongiosis, diverticula likely fall within the range of a normal anatomic variation. Familiarity with these findings may prevent neuroradiologists from recommending unnecessary additional testing in pediatric patients with isolated internal auditory canal diverticula.

In situ phylogenetic structure and diversity of wild Bradyrhizobium communities.

Bacteria often infect their hosts from environmental sources, but little is known about how environmental and host-infecting populations are related. Here, phylogenetic clustering and diversity were investigated in a natural community of rhizobial bacteria from the genus Bradyrhizobium. These bacteria live in the soil and also form beneficial root nodule symbioses with legumes, including those in the genus Lotus. Two hundred eighty pure cultures of Bradyrhizobium bacteria were isolated and genotyped from wild hosts, including Lotus angustissimus, Lotus heermannii, Lotus micranthus, and Lotus strigosus. Bacteria were cultured directly from symbiotic nodules and from two microenvironments on the soil-root interface: root tips and mature (old) root surfaces. Bayesian phylogenies of Bradyrhizobium isolates were reconstructed using the internal transcribed spacer (ITS), and the structure of phylogenetic relatedness among bacteria was examined by host species and microenvironment. Inoculation assays were performed to confirm the nodulation status of a subset of isolates. Most recovered rhizobial genotypes were unique and found only in root surface communities, where little bacterial population genetic structure was detected among hosts. Conversely, most nodule isolates could be classified into several related, hyper-abundant genotypes that were phylogenetically clustered within host species. This pattern suggests that host infection provides ample rewards to symbiotic bacteria but that host specificity can strongly structure only a small subset of the rhizobial community.

Glacial surface temperatures of the southeast Atlantic Ocean.

A detailed record of sea surface temperature from sediments of the Cape Basin in the subtropical South Atlantic indicates a previously undocumented progression of marine climate change between 41 and 18 thousand years before the present (ky B.P.), during the last glacial period. Whereas marine records typically indicate a long-term cooling into the Last Glacial Maximum (around 21 ky B.P.) consistent with gradually increasing global ice volume, the Cape Basin record documents an interval of substantial temperate ocean warming from 41 to 25 ky B.P. The pattern is similar to that expected in response to changes in insolation owing to variations in Earth's tilt.

Competitive effects of some cations on active potassium transport in the human red blood cell.

The effect of some cations on the active potassium transport system of the human red blood cell has been investigated. At low extracellular potassium concentrations, extracellular sodium competitively inhibits the active potassium influx at all sodium concentrations investigated, and tetraethylammonium behaves in a fashion similar to that of sodium. At low extracellular concentrations of potassium, ammonium at low concentrations at first stimulates the active potassium influx, but at higher concentrations inhibits it. Tetramethylammonium at most slightly stimulates the active potassium influx, and calcium is without effect. The behavior is consistent with a model in which potassium is required at more than one site before transport occurs, and the sites are indistinguishable as far as their behavior toward the ions investigated is concerned. The affinity of the alkali metal cations for the sites appears to be explicable in terms of their physical characteristics.

Recoupling the Na-K pump.

Human red blood cells display under appropriate circumstances a ouabain-sensitive K-K exchange when the flux measurements are made using radioisotopes. Such an exchange complicates measurements of the coupling of Na outflux to K influx in cells which are partially depleted of energy sources by deprivation of glucose since the K-K exchange has been found to be increased in depleted cells. When the measurements of flux are made by estimating net cation movements chemically, it is found that glucose deprivation results in a fall in both ouabain-sensitive Na outflux and ouabain-sensitive K influx. Since both fluxes fell in concert, there is no reason for believing that the fluxes are not coupled or that the source of ATP for the Na outflux is different from that for the K influx.

The concentration dependence of active potassium transport in the human red blood cell.

The relation between the active potassium influx in the human red blood cell and the extracellular potassium concentration does not appear to be consistent with the Michaelis-Menten model, but is adequately described by a model in which two potassium ions are required simultaneously at some site or sites in the transport mechanism before transport occurs. The same type of relation appears to exist between that portion of the sodium outflux that requires the presence of extracellular potassium and the extracellular potassium concentration. Rubidium, cesium, and lithium, which are apparently transported by the same system that transports potassium, stimulate the potassium influx when both potassium and the second ion are present at low concentrations, as is predicted by the two-site model.

Concentration dependence of active potassium transport in the human red blood cell in the presence of inhibitors.

The active potassium influx in the human red blood cell is inhibited by strophanthidin, ethacrynic acid, and MK-870 (a new diuretic), and the degree of inhibition is greater at low concentrations of extracellular potassium than at high. In the case of ethacrynic acid, potassium appears to diminish the rate of combination of the drug with the transport system. The kinetic behavior of the active potassium influx in the presence of the inhibitors strophanthidin and ethacrynic acid is consistent with a model in which the binding of potassium at one of the potassium-sensitive sites in the transport system reduces the affinity of the system for the drug, and binding of a second potassium ion further reduces the affinity. It is not possible to distinguish between the sites on the basis of the studies presented here.

Using Model-Based "Learn and Confirm" to Reveal the Pharmacokinetics-Pharmacodynamics Relationship of Pembrolizumab in the KEYNOTE-001 Trial.

Evaluation of pharmacokinetic/pharmacodynamic (PK/PD) properties played an important role in the early clinical development of pembrolizumab. Because analysis of data from a traditional 3 + 3 dose-escalation design revealed several critical uncertainties, a model-based approach was implemented to better understand these properties. Based on anticipated scenarios for potency and PK nonlinearity, a follow-up study was designed and thoroughly evaluated. Execution of 14,000 virtual trials led to the selection and implementation of a robust design that extended the low-dose range by 200-fold. Modeling of the resulting data demonstrated that pembrolizumab PKs are nonlinear at <0.3 mg/kg every 3 weeks, but linear in the clinical dose range. Saturation of ex vivo target engagement in blood began at ≥1 mg/kg every 3 weeks, and a steady-state dose of 2 mg/kg every 3 weeks was needed to reach 95% target engagement, supporting examination of 2 mg/kg every 3 weeks in ongoing trials in melanoma and other advanced cancers.

An empirical test of partner choice mechanisms in a wild legume-rhizobium interaction.

Mutualisms can be viewed as biological markets in which partners of different species exchange goods and services to their mutual benefit. Trade between partners with conflicting interests requires mechanisms to prevent exploitation. Partner choice theory proposes that individuals might foil exploiters by preferentially directing benefits to cooperative partners. Here, we test this theory in a wild legumerhizobium symbiosis. Rhizobial bacteria inhabit legume root nodules and convert atmospheric dinitrogen (N2) to a plant available form in exchange for photosynthates. Biological market theory suits this interaction because individual plants exchange resources with multiple rhizobia. Several authors have argued that microbial cooperation could be maintained if plants preferentially allocated resources to nodules harbouring cooperative rhizobial strains. It is well known that crop legumes nodulate non-fixing rhizobia, but allocate few resources to those nodules. However, this hypothesis has not been tested in wild legumes which encounter partners exhibiting natural, continuous variation in symbiotic benefit. Our greenhouse experiment with a wild legume, Lupinus arboreus, showed that although plants frequently hosted less cooperative strains, the nodules occupied by these strains were smaller. Our survey of wild-grown plants showed that larger nodules house more Bradyrhizobia, indicating that plants may prevent the spread of exploitation by favouring better cooperators.

The role of (alpha beta) protomer interaction in determining functional characteristics of red cell Na,K-ATPase.

We have examined the possibility that interaction of (alpha beta) protomers within a diprotomer is responsible for some anomalous characteristics of red cell Na,K-ATPase by examining their response to two inhibitors, FITC and H2DIDS, which bind covalently, and to ouabain, which debinds slowly from red cell pumps. The phenomena we examined were: (1) the biphasic curve relating Na,K-ATPase activity to ATP concentration, and (2) protection of Na pumps against vanadate inhibition by external Na. If interaction of (alpha beta) protomers within a diprotomer were responsible for these phenomena, random inactivation of (alpha beta) protomers should have resulted in a high proportion of (alpha beta) promtomers with an inhibited protomer as a partner, and therefore should have significantly altered the consequences of subunit interaction. With each inhibitor, 60-70% inhibition of ATPase activity did not alter the functional characteristics of the residual activity. We conclude that interaction of functional (alpha beta) protomers does not explain the phenomena which we investigated. This is consistent with our previous observation that Na,K pumps of red cell membranes exist as monomeric (alpha beta) protomers (Martin, D.W. and Sachs, V.R. (1992) J. Biol. Chem. 267, 23922-23929).