Harmonizing spatial scales and ecological theories to predict avian richness and functional diversity within forest ecosystems.

Classic ecological theory has proven that temperature, precipitation and productivity organize ecosystems at broad scales and are generalized drivers of biodiversity within different biomes. At local scales, the strength of these predictors is not consistent across different biomes. To better translate these theories to localized scales, it is essential to determine the links between drivers of biodiversity. Here we harmonize existing ecological theories to increase the predictive power for species richness and functional diversity. We test the relative importance of three-dimensional habitat structure as a link between local and broad-scale patterns of avian richness and functional diversity. Our results indicate that habitat structure is more important than precipitation, temperature and elevation gradients for predicting avian species richness and functional diversity across different forest ecosystems in North America. We conclude that forest structure, influenced by climatic drivers, is essential for predicting the response of biodiversity with future shifts in climatic regimes.

Effects of forest fragmentation on avian breeding activity.

Biodiversity declines and ecosystem decay follow forest fragmentation; initially, abundant species may become rare or be extirpated. Underlying mechanisms behind delayed extirpation of certain species following forest fragmentation are unknown. Species declines may be attributed to an inadequate number of breeding adults required to replace the population or decreased juvenile survival rate due to reduced recruitment or increased nest predation pressures. We used 10 years of avian banding data, 5 years before and 4 years after fragment isolation, from the Biological Dynamics of Forest Fragments Project, carried out near Manaus, Brazil, to investigate the breeding activity hypothesis that there is less breeding activity and fewer young after relative to before fragment isolation. We compared the capture rates of active breeding and young birds in 3 forest types (primary forest, fragment before isolation, and fragment after isolation) and the proportion of active breeding and young birds with all birds in each unique fragment type before and after isolation. We grouped all bird species by diet (insectivore or frugivore) and nesting strategy (open cup, cavity, or enclosed) to allow further comparisons among forest types. We found support for the breeding activity hypothesis in insectivorous and frugivorous birds (effect sizes 0.45 and 0.53, respectively) and in birds with open-cup and enclosed nesting strategies (effect sizes 0.56 and 0.44, respectively) such that on average there were more breeding birds in fragments before isolation relative to after isolation. A larger proportion of birds in the community were actively breeding before fragment isolation (72%) than after fragment isolation (11%). Unexpectedly, there was no significant decrease in the number of young birds after fragment isolation, although sample sizes for young were small (n = 43). This may have been due to sustained immigration of young birds to fragments after isolation. Together, our results provide some of the strongest evidence to date that avian breeding activity decreases in response to fragment isolation, which could be a fundamental mechanism contributing to ecosystem decay.

Efectos de la fragmentación del bosque sobre la actividad reproductiva de las aves Resumen Les declinaciones de la biodiversidad y el deterioro de los ecosistemas van después de la fragmentación forestal; al inicio, las especies abundantes pueden volverse raras o ser extirpadas. Todavía no se conocen los mecanismos subyacentes detrás de la extirpación retrasada de ciertas especies después de la fragmentación forestal. La declinación de las especies puede atribuirse a un número inadecuado de adultos reproductivos requeridos para reemplazar a la población o a la tasa reducida de supervivencia de los juveniles debido al reclutamiento disminuido o al incremento en la presión de depredación de los nidos. Usamos diez años de datos de anillamiento de aves, cinco años antes y cuatro años después del aislamiento por fragmentación, tomados del Proyecto Dinámica Biológica de Fragmentos de Bosque realizado cerca de Manaos, Brasil, para investigar la hipótesis de actividad reproductiva que sostiene que hay una menor actividad reproductiva y menos crías después del aislamiento por fragmentación que antes del aislamiento. Comparamos las tasas de captura de aves con reproducción activa y aves juveniles en tres tipos de bosque (primario, fragmento antes del aislamiento y fragmento después del aislamiento) y la proporción de las aves juveniles y con reproducción activa con todas las aves en cada tipo de fragmento único antes y después del aislamiento. Agrupamos todas las especies de aves según su dieta (insectívora o frugívora) y su estrategia de anidación (nido abierto, cavidad o nido cerrado) para poder realizar más comparaciones entre los tipos de bosque. Las aves con dieta insectívora y frugívora (tamaño del efecto: 0.45 y 0.53, respectivamente) y aquellas con nidos abiertos y cerrados (tamaño del efecto: 0.56 y 0.44, respectivamente) respaldaron la hipótesis de la actividad reproductora de tal manera que en promedio hubo más aves reproductoras en los fragmentos antes del aislamiento que después del aislamiento. Una gran parte de las aves de la comunidad tuvieron reproducción activa antes del aislamiento por fragmentación (72%) que después del aislamiento (11%). Sorprendentemente, no hubo una disminución significativa en el número de aves juveniles después del aislamiento, si bien el tamaño de la muestra de este grupo fue reducido (n = 43). Lo anterior pudo deberse a la continua inmigración de juveniles a los fragmentos después del aislamiento. En conjunto, nuestros resultados proporcionan algunos de los indicios más claros de que la actividad reproductiva de las aves disminuye como respuesta al aislamiento por fragmentación, lo cual podría ser un mecanismo fundamental del deterioro de los ecosistemas.

Acquisition and Transmission of 'Candidatus Liberibacter solanacearum' Differs Among Wolbachia-Infected and -Uninfected Haplotypes of Bactericera cockerelli.

'Candidatus Liberibacter solanacearum' (Lso) causes disease symptoms and economic losses in potato, tomato, and other solanaceous crops in North America. Lso is transmitted to plants by the potato psyllid, Bactericera cockerelli, which occurs as distinct haplotypes named western, central, and northwestern that differ in the presence or absence of the bacterial endosymbiont, Wolbachia. Previous work showed that all three vector haplotypes can transmit Lso, but it was not clear whether acquisition and transmission rates of Lso were equal among the haplotypes. The goal of our study was to compare Lso infection rates among psyllids of the western, central, and northwestern haplotypes. Using data collected from several years of periodic testing of Lso infection of laboratory-reared potato psyllid colonies, we showed that psyllids of the western and central haplotypes are more likely to harbor Lso than are psyllids of the northwestern haplotype. We then used greenhouse assays to demonstrate that psyllids of the northwestern haplotype are less likely to acquire and transmit Lso than those of the western haplotype. Lso infection rates corresponded with Wolbachia infection among the three psyllid haplotypes. The Wolbachia-infected central and western haplotypes were more likely to harbor and transmit Lso than the Wolbachia-free northwestern haplotype. Results demonstrate that potato psyllids of the western and central haplotypes pose a greater risk for spread of Lso in crops and suggest a pattern between infection with Lso and Wolbachia in potato psyllid.

Do adult Magicicada (Hemiptera: Cicadidae) feed? Historical perspectives and evidence from molecular gut content analysis.

The periodical cicadas in the genus Magicicada are remarkable for their unusual life histories and dramatic synchronized emergences every 13 or 17 years. While aspects of their evolution, mating behaviors, and general biology have been well-characterized, there is surprising uncertainty surrounding the feeding habits of the short-lived adult stage. Despite a tentative scientific consensus to the contrary, the perception that adult Magicicada do not feed has persisted among the general public, and recent studies are lacking. We directly investigated the feeding behavior of Magicicada spp. through high-throughput sequencing (HTS)-based dietary analysis of nymphs, freshly molted (teneral) adults, and fully sclerotized adults collected from orchard and wooded habitats during the 2021 emergence of Brood X. Identifiable plant DNA (trnF, ITS amplicons) was successfully recovered from nymphs and adults. No plant DNA was recovered from teneral adults, suggesting that all DNA recovered from sclerotized adults was ingested during the post-teneral adult stage. Both nymphs and adults were found to have ingested a range of woody and herbaceous plants across 17 genera and 14 families. Significantly more plant genera per individual were recovered from adults than from nymphs, likely reflecting the greater mobility of the adult stage. We hypothesize that the demonstrated ingestion of plant sap by Magicicada adults is driven by a need to replace lost water and support specialized bacteriome-dwelling endosymbionts that cicadas depend upon for growth and development, which constitutes true feeding behavior.

Residual ctDNA after treatment predicts early relapse in patients with early-stage non-small cell lung cancer.

BACKGROUND: Identification of residual disease in patients with localized non-small cell lung cancer (NSCLC) following treatment with curative intent holds promise to identify patients at risk of relapse. New methods can detect circulating tumour DNA (ctDNA) in plasma to fractional concentrations as low as a few parts per million, and clinical evidence is required to inform their use. PATIENTS AND METHODS: We analyzed 363 serial plasma samples from 88 patients with early-stage NSCLC (48.9%/28.4%/22.7% at stage I/II/III), predominantly adenocarcinomas (62.5%), treated with curative intent by surgery (n = 61), surgery and adjuvant chemotherapy/radiotherapy (n = 8), or chemoradiotherapy (n = 19). Tumour exome sequencing identified somatic mutations and plasma was analyzed using patient-specific RaDaR™ assays with up to 48 amplicons targeting tumour-specific variants unique to each patient. RESULTS: ctDNA was detected before treatment in 24%, 77% and 87% of patients with stage I, II and III disease, respectively, and in 26% of all longitudinal samples. The median tumour fraction detected was 0.042%, with 63% of samples <0.1% and 36% of samples <0.01%. ctDNA detection had clinical specificity >98.5% and preceded clinical detection of recurrence of the primary tumour by a median of 212.5 days. ctDNA was detected after treatment in 18/28 (64.3%) of patients who had clinical recurrence of their primary tumour. Detection within the landmark timepoint 2 weeks to 4 months after treatment end occurred in 17% of patients, and was associated with shorter recurrence-free survival [hazard ratio (HR): 14.8, P <0.00001] and overall survival (HR: 5.48, P <0.0003). ctDNA was detected 1-3 days after surgery in 25% of patients yet was not associated with disease recurrence. Detection before treatment was associated with shorter overall survival and recurrence-free survival (HR: 2.97 and 3.14, P values 0.01 and 0.003, respectively). CONCLUSIONS: ctDNA detection after initial treatment of patients with early-stage NSCLC using sensitive patient-specific assays has potential to identify patients who may benefit from further therapeutic intervention.

Long-term changes in avian biomass and functional diversity within disturbed and undisturbed Amazonian rainforest.

Recent long-term studies in protected areas have revealed the loss of biodiversity, yet the ramifications for ecosystem health and resilience remain unknown. Here, we investigate how the loss of understory birds, in the lowest stratum of the forest, affects avian biomass and functional diversity in the Amazon rainforest. Across approximately 30 years in the Biological Dynamics of Forest Fragments Project, we used a historical baseline of avian communities to contrast the avian communities in today's primary forest with those in modern disturbed habitat. We found that in primary rainforest, the reduced abundance of insectivorous species led to reduced functional diversity, but no reduction of biomass, indicating that species with similar functional traits are less likely to coexist in modern primary forests. Because today's forests contain fewer functionally redundant species-those with similar traits-we argue that avian communities in modern primary Amazonian rainforests are less resilient, which may ultimately disrupt the ecosystem in dynamic and unforeseen ways.

A new picorna-like virus identified in populations of the potato psyllid Bactericera cockerelli.

The potato/tomato psyllid Bactericera cockerelli (Hemiptera: Triozidae) is a pest of Solanaceae plants and a vector of the pathogenic bacterium 'Candidatus Liberibacter solanacearum', which is associated with zebra chip disease in potato. This disease is controlled through insecticide treatments, and more environmentally friendly management options are desirable. The objective of this study was to identify viruses present in potato psyllid populations that might be used as biocontrol agents for this insect pest. A new picorna-like virus, tentatively named "Bactericera cockerelli picorna-like virus" (BcPLV), was discovered in B. cockerelli populations maintained in greenhouses, through the use of high-throughput sequencing data and subsequent confirmation by RT-PCR and Sanger sequencing. BcPLV has a positive-sense 9,939-nt RNA genome encoding a single 2,947-aa polyprotein and is related to the Diaphorina citri picorna-like virus (DcPLV) found in Asian citrus psyllid Diaphorina citri populations. Based on their genome organization and the phylogeny of their RNA-dependent RNA polymerase domains, BcPLV and DcPLV together are proposed to comprise a new genus, provisionally named "Psylloidivirus", within the family Iflaviridae.

Myositis ossificans: a rare neonatal presentation.

Myositis ossificans is a benign, ossifying, soft-tissue pseudotumor that most commonly occurs in men ages 30-40 years after trauma. Myositis ossificans may also occur in children, but it is extremely rare in those younger than 10 years of age. While myositis ossificans can often mimic malignant soft-tissue tumors, it has many unique findings that can aid in diagnostic differentiation. This differentiation is critical to avoid unnecessary risk with potentially harmful procedures. We present a very unusual presentation of myositis ossificans in the immediate post-birth perinatal period, as well as a review of key imaging findings.

'Candidatus Liberibacter solanacearum' Infection of Physalis ixocarpa Brot. (Solanales: Solanaceae) in Saltillo, Mexico.

The potato psyllid Bactericera cockerelli (Sulc) (Hemiptera: Triozidae) is a pest of solanaceous crops (order Solanales), including potato (Solanum tuberosum L.) and tomato (S. lycopersicum L.). Feeding by high populations of nymphs causes psyllid yellows while adults and nymphs are vectors of the plant pathogen 'Candidatus Liberibacter solanacearum'. Foliar symptoms that were consistent with either 'Ca. L. solanacearum' infection or psyllid yellows were observed in 2019 on tomatillo (Physalis ixocarpa Brot.; family Solanaceae) grown within an experimental plot located near Saltillo, Mexico. This study had three primary objectives: 9i) determine whether the foliar symptoms observed on tomatillo were associated with 'Ca. L. solanacearum' infection, (ii) identify the haplotypes of 'Ca. L. solanacearum' and potato psyllids present in the symptomatic plot, and (iii) use gut content analysis to infer the plant sources of 'Ca. L. solanacearum'-infected psyllids. Results confirmed that 71% of symptomatic plants and 71% of psyllids collected from the plants were infected with 'Ca. L. solanacearum'. The detection of 'Ca. L. solanacearum' in plants and psyllids and the lack of nymphal populations associated with psyllid yellows strongly suggests that the observed foliar symptoms were caused by 'Ca. L. solanacearum' infection. All infected plants and insects harbored the more virulent 'Ca. L. solanacearum' haplotype B but one psyllid was also coinfected with haplotype A. The potato psyllids were predominantly of the central haplotype but one psyllid was identified as the western haplotype. Molecular gut content analysis of psyllids confirmed the movement of psyllids between noncrop habitats and tomatillo and indicated that 'Ca. L. solanacearum' infection of psyllids was associated with increased plant diversity in their diet.

Functional morphogenesis from embryos to adults: Late development shapes trophic niche in coral reef damselfishes.

The damselfishes are one of the dominant coral reef fish lineages. Their ecological diversification has involved repeated transitions between pelagic feeding using fast bites and benthic feeding using forceful bites. A highly-integrative approach that combined gene expression assays, shape analyses, and high-speed video analyses was used to examine the development of trophic morphology in embryonic, larval, juvenile, and adult damselfishes. The anatomical characters that distinguish pelagic-feeding and benthic-feeding species do not appear until after larval development. Neither patterns of embryonic jaw morphogenesis, larval skull shapes nor larval bite mechanics significantly distinguished damselfishes from different adult trophic guilds. Analyses of skull shape and feeding performance identified two important transitions in the trophic development of a single species (the orange clownfish; Amphiprion percula): (a) a pronounced transformation in feeding mechanics during metamorphosis; and (b) more protracted cranial remodeling over the course of juvenile development. The results of this study indicate that changes in postlarval morphogenesis have played an important role in damselfish evolution. This is likely to be true for other fish lineages, particularly if they consist of marine species, the majority of which have planktonic larvae with different functional requirements for feeding in comparison to their adult forms.

Identification of an Optimized Heating and Fast Ion Generation Scheme for the Wendelstein 7-X Stellarator.

A Doppler shifted resonance minority species ion cyclotron range of frequency (ICRF) scheme for heating neutral beam ions has been identified and optimized for the Wendelstein 7-X stellarator. Compared with more conventional methods, the synergetic scheme increases the normalized core collisional power transfer to the background plasma, and induces larger concentrations of energetic ions. Simulations in the intricate 3D magnetic stellarator geometry reveal an energetic distribution function that is only weakly anisotropic, and is thus relevant to fast ion and alpha particle driven Alfvén eigenmode experimental preparation. Quasilinear theory and simulations of the Joint European Torus indicate that the excellent confinement properties are due to increased velocity diffusion from ICRF interaction along the magnetic field lines. Agreement is found between SCENIC simulations and Joint European Torus experimental measurements for the total neutron rate and the energy distribution of the fast ions.

Growth and Yield Performance of Solanum tuberosum Grown from Seed Potatoes Infected with 'Candidatus Liberibacter solanacearum' Haplotypes A and B.

Zebra chip (ZC) disease of potato (Solanum tuberosum) is associated with infection by 'Candidatus Liberibacter solanacearum' (Lso). Two haplotypes of Lso-A and B-occur in the United States. Lso haplotype B is more virulent than haplotype A, causing greater disease incidence in tubers, more severe symptoms, and greater loss in tuber yield. This study assessed whether tubers from infected plants generate new infected plants the following year. The effects of both Lso haplotypes A and B on tuber resprout were examined on five potato cultivars. When compared with noninfected tubers, overall plant emergence rate from Lso A- or B-infected tubers was lower, plants emerged slower, and plants generated lower daughter tuber yields in weight and quantity. Plants generally emerged poorly from Lso B-infected tubers and produced lower daughter tuber yields than Lso A-infected tubers. Regardless of Lso treatment, all daughter tubers were asymptomatic, and only 0.3% tested positive for Lso in experiments conducted over 2 years. This suggests that plants generated from Lso A- and Lso B-infected seed potatoes with severe ZC symptoms are likely not a significant source of Lso in potato fields.

Thyroid hormone modulation during zebrafish development recapitulates evolved diversity in danionin jaw protrusion mechanics.

Protrusile jaws are a highly useful innovation that has been linked to extensive diversification in fish feeding ecology. Jaw protrusion can enhance the performance of multiple functions, such as suction production and capturing elusive prey. Identifying the developmental factors that alter protrusion ability will improve our understanding of fish diversification. In the zebrafish protrusion arises postmetamorphosis. Fish metamorphosis typically includes significant changes in trophic morphology, accompanies a shift in feeding niche and coincides with increased thyroid hormone production. We tested whether thyroid hormone affects the development of zebrafish feeding mechanics. We found that it affected all developmental stages examined, but that effects were most pronounced after metamorphosis. Thyroid hormone levels affected the development of jaw morphology, feeding mechanics, shape variation, and cranial ossification. Adult zebrafish utilize protrusile jaws, but an absence of thyroid hormone impaired development of the premaxillary bone, which is critical to jaw protrusion. Premaxillae from early juvenile zebrafish and hypothyroid adult zebrafish resemble those from adults in the genera Danionella, Devario, and Microdevario that show little to no jaw protrusion. Our findings suggest that evolutionary changes in how the developing skulls of danionin minnows respond to thyroid hormone may have promoted diversification into different feeding niches.

Excitation Mechanism of Low-n Edge Harmonic Oscillations in Edge Localized Mode-Free, High Performance, Tokamak Plasmas.

The excitation mechanism for low-n edge harmonic oscillations in quiescent H-mode regimes is identified analytically. We show that the combined effect of diamagnetic and poloidal magnetohydrodynamic flows, with the constraint of a Doppler-like effect of the ion flow, leads to the stabilization of short wavelength modes, allowing low-n perturbation to grow. The analysis, performed in tokamak toroidal geometry, includes the effects of large edge pressure gradients, associated with the local flattening of the safety factor and diamagnetic flows, sheared parallel and E×B rotation, and a vacuum region between plasma and the ideal metallic wall. The separatrix also is modeled analytically.

How phylogeny and foraging ecology drive the level of chemosensory exploration in lizards and snakes.

The chemical senses are crucial for squamates (lizards and snakes). The extent to which squamates utilize their chemosensory system, however, varies greatly among taxa and species' foraging strategies, and played an influential role in squamate evolution. In lizards, 'Scleroglossa' evolved a state where species use chemical cues to search for food (active foragers), whereas 'Iguania' retained the use of vision to hunt prey (ambush foragers). However, such strict dichotomy is flawed as shifts in foraging modes have occurred in all clades. Here, we attempted to disentangle effects of foraging ecology from phylogenetic trait conservatism as leading cause of the disparity in chemosensory investment among squamates. To do so, we used species' tongue-flick rate (TFR) in the absence of ecological relevant chemical stimuli as a proxy for its fundamental level of chemosensory investigation, that is baseline TFR. Based on literature data of nearly 100 species and using phylogenetic comparative methods, we tested whether and how foraging mode and diet affect baseline TFR. Our results show that baseline TFR is higher in active than ambush foragers. Although baseline TFRs appear phylogenetically stable in some lizard taxa, that is a consequence of concordant stability of foraging mode: when foraging mode shifts within taxa, so does baseline TFR. Also, baseline TFR is a good predictor of prey chemical discriminatory ability, as we established a strong positive relationship between baseline TFR and TFR in response to prey. Baseline TFR is unrelated to diet. Essentially, foraging mode, not phylogenetic relatedness, drives convergent evolution of similar levels of squamate chemosensory investigation.

The evolution of jaw protrusion mechanics is tightly coupled to bentho-pelagic divergence in damselfishes (Pomacentridae).

Most species-rich lineages of aquatic organisms have undergone divergence between forms that feed from the substrate (benthic feeding) and forms that feed from the water column (pelagic feeding). Changes in trophic niche are frequently accompanied by changes in skull mechanics, and multiple fish lineages have evolved highly specialized biomechanical configurations that allow them to protrude their upper jaws toward the prey during feeding. Damselfishes (family Pomacentridae) are an example of a species-rich lineage with multiple trophic morphologies and feeding ecologies. We sought to determine whether bentho-pelagic divergence in the damselfishes is tightly coupled to changes in jaw protrusion ability. Using high-speed video recordings and kinematic analysis, we examined feeding performance in 10 species that include three examples of convergence on herbivory, three examples of convergence on omnivory and two examples of convergence on planktivory. We also utilized morphometrics to characterize the feeding morphology of an additional 40 species that represent all 29 damselfish genera. Comparative phylogenetic analyses were then used to examine the evolution of trophic morphology and biomechanical performance. We find that pelagic-feeding damselfishes (planktivores) are strongly differentiated from extensively benthic-feeding species (omnivores and herbivores) by their jaw protrusion ability, upper jaw morphology and the functional integration of upper jaw protrusion with lower jaw abduction. Most aspects of cranial form and function that separate these two ecological groups have evolved in correlation with each other and the evolution of the functional morphology of feeding in damselfishes has involved repeated convergence in form, function and ecology.

Elicitors of Host Plant Defenses Partially Suppress Cacopsylla pyricola (Hemiptera: Psyllidae) Populations Under Field Conditions.

Defense elicitors are products that activate acquired defense responses in plants, thus rendering the plants less susceptible to attack by a broad range of pests. We demonstrated previously under laboratory conditions that foliar applications of the defense elicitors Actigard (acibenzolar-S-methyl), Employ (harpin protein), or ODC (chitosan) to potted pear trees (Pyrus communis L.) each caused an increase in mortality of Cacopsylla pyricola (Förster) (Hemiptera: Psyllidae) nymphs and altered the settling and oviposition behavior of the adults. In this study, we monitored C. pyricola populations over a 3-yr period on orchard-grown trees treated with water (untreated control), Actigard, Employ, or ODC. Fewer nymphs were observed on trees treated with elicitors compared with untreated trees in both 2014 and 2016. A similar but statistically nonsignificant pattern was observed in 2015 when nearly 30% fewer nymphs were observed on trees treated with elicitors versus untreated controls. Observed reductions in psyllid numbers by defense elicitors were modest and do not warrant the use of these products alone for managing C. pyricola. However, these products are often used for management of fire blight, and our observations that elicitors also reduce C. pyricola populations may be useful for system-wide integrated pest management approaches.

Relative Abundance of Carsonella ruddii (Gamma Proteobacterium) in Females and Males of Cacopsylla pyricola (Hemiptera: Psyllidae) and Bactericera cockerelli (Hemiptera: Triozidae).

Carsonella ruddii (Gamma Proteobacterium) is an obligate bacterial endosymbiont of psyllids that produces essential amino acids that are lacking in the insect's diet. Accurate estimations of Carsonella populations are important to studies of Carsonella-psyllid interactions and to developing ways to target Carsonella for control of psyllid pests including pear psylla, Cacopsylla pyricola (Förster) (Hemiptera: Psyllidae) and potato psyllid, Bactericera cockerelli (Sulc) (Hemiptera: Triozidae). We used two methods, namely fluorescence in situ hybridization and quantitative polymerase chain reaction (qPCR), to estimate relative abundance of Carsonella in bacteriocytes and whole bodies of psyllids, respectively. Using these two methods, we compared Carsonella populations between female and male insects. Estimations using fluorescence in situ hybridization indicated that Carsonella was more abundant in bacteriocytes of female C. pyricola than in those of males, but Carsonella abundance in bacteriocytes did not differ between sexes of B. cockerelli. Analyses by qPCR using whole-body specimens indicated Carsonella was more abundant in females than in males of both psyllids. Neither fluorescence in situ hybridization nor qPCR indicated that Carsonella populations differed in abundance among adults of different ages (0-3 wk after adult eclosion). Using fluorescence in situ hybridization, Carsonella was observed in ovarioles of newly emerged females and formed an aggregation in the posterior end of mature oocytes. Results of our study indicate that female psyllids harbor greater populations of Carsonella than do males and that sex should be controlled for in studies which require estimations of Carsonella populations.

Correlation of BRAF and NRAS mutation status with outcome, site of distant metastasis and response to chemotherapy in metastatic melanoma.

BACKGROUND: The prognostic significance of BRAF and NRAS mutations in metastatic melanoma patients remains uncertain, with several studies reporting conflicting results, often biased by the inclusion of patients treated with BRAF and MEK (MAPK) inhibitors. We therefore interrogated a historical cohort of patients free of the confounding influence of MAPK inhibitor therapy. METHODS: Patients with available archival tissue first diagnosed with metastatic melanoma between 2002 and 2006 were analysed. Mutational analysis was performed using the OncoCarta Panel. Patient characteristics, treatment outcome and survival were correlated with BRAF/NRAS mutation status. RESULTS: In 193 patients, 92 (48%) melanomas were BRAF-mutant, 39 (20%) were NRAS-mutant and 62 (32%) were wild-type for BRAF/NRAS mutations (wt). There was no difference in response to chemotherapy based on mutation status (35-37%). The distant disease-free interval (DDFI) was significantly shorter in patients with wt melanoma (27.9 months vs 35.1 for BRAF and 49.1 for NRAS) although this was not significant in multivariate analysis. Survival from stage IV melanoma diagnosis was not significantly different based on mutation status. The DDFI was significantly shorter in patients with BRAF(V600K/R) versus BRAF(V600E) melanoma in univariate and multivariate analyses. CONCLUSIONS: BRAF and NRAS mutation status does not influence survival in metastatic melanoma.

Fat-suppression techniques for 3-T MR imaging of the musculoskeletal system.

Fat suppression is an important technique in musculoskeletal imaging to improve the visibility of bone-marrow lesions; evaluate fat in soft-tissue masses; optimize the contrast-to-noise ratio in magnetic resonance (MR) arthrography; better define lesions after administration of contrast material; and avoid chemical shift artifacts, primarily at 3-T MR imaging. High-field-strength (eg, 3-T) MR imaging has specific technical characteristics compared with lower-field-strength MR imaging that influence the use and outcome of various fat-suppression techniques. The most commonly used fat-suppression techniques for musculoskeletal 3-T MR imaging include chemical shift (spectral) selective (CHESS) fat saturation, inversion recovery pulse sequences (eg, short inversion time inversion recovery [STIR]), hybrid pulse sequences with spectral and inversion-recovery (eg, spectral adiabatic inversion recovery and spectral attenuated inversion recovery [SPAIR]), spatial-spectral pulse sequences (ie, water excitation), and the Dixon techniques. Understanding the different fat-suppression options allows radiologists to adopt the most appropriate technique for their clinical practice.