PD-L1 expression in nonclear-cell renal cell carcinoma.

BACKGROUND: Programmed death ligand-1 (PD-L1) expression in nonclear-cell RCC (non-ccRCC) and its association with clinical outcomes are unknown. METHODS: Formalin-fixed paraffin-embedded (FFPE) specimens were obtained from 101 patients with non-ccRCC. PD-L1 expression was evaluated by immunohistochemistry in both tumor cell membrane and tumor-infiltrating mononuclear cells (TIMC). PD-L1 tumor positivity was defined as ≥5% tumor cell membrane staining. For PD-L1 expression in TIMC, a combined score based on the extent of infiltrate and percentage of positive cells was used. Baseline clinico-pathological characteristics and outcome data [time to recurrence (TTR) and overall survival (OS)] were correlated with PD-L1 staining. RESULTS: Among 101 patients, 11 (10.9%) were considered PD-L1+ in tumor cells: 2/36 (5.6%) of chromophobe RCC, 5/50 (10%) of papillary RCC, 3/10 (30%) of Xp11.2 translocation RCC and 1/5 (20%) of collecting duct carcinoma. PD-L1 positivity (PD-L1+) in tumor cells was significantly associated with higher stage (P = 0.01) and grade (P = 0.03), as well as shorter OS (P < 0.001). On the other hand, PD-L1 positivity by TIMC was observed in 57 (56.4%) patients: 13/36 (36.1%) of chromophobe RCC, 30/50 (60%) of papillary RCC, 9/10 (90%) of Xp11.2 translocation RCC and 5/5 (100%) of collecting duct carcinoma. A trend toward shorter OS was observed in patients with PD-L1+ in TIMC (P = 0.08). PD-L1+ in both tumor cell membrane and TIMC cells were associated with shorter TTR (P = 0.02 and P = 0.03, respectively). CONCLUSION: In non-ccRCC, patients with PD-L1+ tumors appear to have worse clinical outcomes, although only PD-L1 positivity in tumor cells is associated with higher tumor stage and grade.

Influence of environmental quality on pollen competitive ability in wild radish.

Pollen of Raphanus raphanistrum produced under low nutrient conditions sired fewer seeds than pollen produced under better conditions when the two types were applied on a stigma together. No difference was seen in single-donor crosses. Male mating success can be strongly influenced by the environmental conditions of pollen-bearing plants, a factor overlooked in studies of plant reproductive biology and in standard quantitative genetic crossing designs, where effects of male parent are equated with heritable genetic variation.

Floral evolution: attractiveness to pollinators increases male fitness.

Because availability of resources often limits seed or fruit set, increased visits by pollinators may not always lead to increases in maternal reproduction. This observation has led evolutionary biologists to hypothesize that a plant's ability to attract pollinators may have its primary impact on male fitness achieved through the fertlization of ovules. This interpretation of angiosperm reproductive ecology is supported by field experiments. Pollinating insects strongly discriminated between two Mendelian petal-color morphs in Raphanus raphanistrum, a widespread, self-incompatible crucifer. In experimental populations composed of petal-color homozygotes. color discrimination by naturally occurring pollinators had no statistically significant effect on relative maternal function (fruit and seed production) in the two morphs. In contrast, yellow-flowered individuals were far more successful as fathers (pollen donors) than were the less visited whites. These results suggest that the evolution of floral signals such as petal color may be driven primarily by selection on male function.

Evolution in stressful environments. I. Phenotypic variability, phenotypic selection, and response to selection in five distinct environmental stresses.

Considerable debate has accompanied efforts to integrate the selective impacts of environmental stresses into models of life-history evolution. This study was designed to determine if different environmental stresses have consistent phenotypic effects on life-history characters and whether selection under different stresses leads to consistent evolutionary responses. We created lineages of a wild mustard (Sinapis arvensis) that were selected for three generations under five stress regimes (high boron, high salt, low light, low water, or low nutrients) or under near-optimal conditions (control). Full-sibling families from the six selection histories were divided among the same six experimental treatments. In that test generation, lifetime plant fecundity and six phenotypic traits were measured for each plant. Throughout this greenhouse study, plants were grown individually and stresses were applied from the early seedling stage through senescence. Although all stresses consistently reduced lifetime fecundity and most size- and growth-related traits, different stresses had contrasting effects on flowering time. On average, stress delayed flowering compared to favorable conditions, although plants experiencing low nutrient stress flowered earliest and those experiencing low light flowered latest. Contrary to expectations of Grime's triangle model of life-history evolution, this ruderal species does not respond phenotypically to poor environments by flowering earlier. Most stresses enhanced the evolutionary potential of the study population. Compared with near-optimal conditions, stresses tended to increase the opportunity for selection as well as phenotypic variance, although both of these quantities were reduced in some stresses. Rather than favoring traits characteristic of stress tolerance, such as slow growth and delayed reproduction, phenotypic selection favored stress-avoidance traits: earlier flowering in all five stress regimes and faster seedling height growth in three stresses. Phenotypic correlations reinforced direct selection on these traits under stress, leading to predicted phenotypic change under stress, but no significant selection in the control environment. As a result of these factors, selection under stress resulted in an evolutionary shift toward earlier flowering. Environmental stresses may drive populations of ruderal plant species like S. arvensis toward a stress-avoidance strategy, rather than toward stress tolerance. Further studies will be needed to determine when selection in stressful environments leads to these alternative life-history strategies.

Consequences of Intraspecific Competition and Environmental Variation for Selection in the Mustard Sinapsis arvensis: Contrasting Ecological and Evolutionary Perspectives.

Alternative models of plant life-history evolution differ in their views of how abiotic stress and competition interact to shape the evolution of plant life-history traits. To address this debate, which crosses traditional boundaries between community ecology and population biology, we grew wild turnip families from three selection histories in a field experiment in which we manipulated conspecific density and sun exposure. Hot spring conditions caused neutral shading to reduce drought stress, resulting in a greater mean and variance for lifetime fertility at low density and greater intensity of competition at high density. The variance in relative fitness among individuals or families was least in partial shade at low density. Prior selection under shade stress in the greenhouse reduced lifetime fitness in the less stressful partial-shade treatment under field conditions. Patterns of selection and predicted trait evolution were more similar between high and low densities than between the two light environments. Partial shade favored the proliferation of large leaves early in development, especially at high density. Selection in the stressful full-sun treatment favored reduced pathogen susceptibility at both densities and early flowering at low density. Because direct selection on traits changed principally in magnitude rather than in direction, genetic correlations for fitness were generally positive between light and density treatments. Greater intraspecific competition led to more rapid predicted trait evolution in the partial-shade environment but not in the stressful full-sun treatment.

Short-term dynamics of an acacia ant community in Laikipia, Kenya.

In monospecific stands of Acacia drepanolobium in Laikipia, Kenya, virtually all but the smallest trees are occupied by one of four species of ants. Although trees are a limiting resource, all four ant species are maintained in this system. Three separate lines of evidence confirm a linear dominance hierarchy among these four ants: (1) experimentally staged conflicts, (2) natural transitions among 1773 tagged trees over a 6-month period, and (3) the average sizes of trees occupied by ants of different species. Short-term dynamics during a drying period reveal that many smaller trees (<1 m) occupied by dominant ants were subsequently abandoned, and that abandoned trees had grown more slowly than those that were not abandoned. Height growth increments over 6 months were generally independent of ant occupant, but increased with tree height. Among taller trees (>1 m), changes in ant occupation congruent with the dominance hierarchy (i.e., transitions from more subordinate ant species to more dominant ant species) occurred on trees that grew faster than average. In contrast, the (less frequent) changes in ant occupation "against" the direction of the dominance hierarchy occurred on trees that grew more slowly than average. Observed correlations between tree vigor and takeover direction suggest that colony growth of dominant ant species is either favored in more productive microhabitats, or that such colonies differentially seek out healthier trees for conquest. Colonies of dominant species may differentially abandon more slowly growing trees during (dry) periods of retrenchment, or suffer higher mortality on these trees. Subordinate ant species appear to move onto these abandoned trees and, to a lesser extent, colonize new recruits in the sapling class. These data reveal that within a simple linear dominance hierarchy, short-term variations exist that may reveal underlying mechanisms associated with coexistence.

Pollination thoroughness and maternal yield regulation in wild radish, Raphanus raphanistrum (Brassicaceae).

Under conditions where resources are limited, there are often negative correlations between components of maternal yield, or between fruit and flower production. Pollination, in turn, may vary among individuals and influence total maternal expenditure. We examined the impact of variation in pollination thoroughness upon yield components and overall plant growth in wild radish (R. raphanistrum) plants grown in the greenhouse. Plants received different pollination treatments in which 0% to 100% of all flowers produced were hand-pollinated. Fruit set was increased by hand-pollination, but rarely exceeded 30%, even when more than 50% of the flowers were pollinated. Plants receiving more thorough pollination or having greater proportion fruit set produced significantly smaller seeds. Seed number per fruit was not influenced by pollination treatment. Mean values of yield components and interactions between components often varied among plants from different maternal families. Increasing pollination thoroughness also resulted in dramatic decreases in flower production. If male fitness is related to flower number, there may be a tradeoff between maternal fecundity and successful pollen export operating at the whole-plant level in this species.

Psychotherapy for persons with craniofacial deformities: can we treat without theory?

In recent years, increasing numbers of experts have recommended that psychological support be available for cleft children and their parents. Few cleft palate centers however offer comprehensive psychological services. This paper presents some conceptual factors which may contribute to the paucity of psychological treatments available to cleft children and their families. Shortcomings in current concepts of emotional dysfunction in cleft children are discussed, and the effects of conceptual confusion on options for psychotherapy are outlined. Suggested directions in psychotherapy research for cleft children are discussed.

Evolution in stressful environments II: adaptive value and costs of plasticity in response to low light in Sinapis arvensis.

Plants possess a remarkable capacity to alter their phenotype in response to the highly heterogeneous light conditions they commonly encounter in natural environments. In the present study with the weedy annual plant Sinapis arvensis, we (a) tested for the adaptive value of phenotypic plasticity in morphological and life history traits in response to low light and (b) explored possible fitness costs of plasticity. Replicates of 31 half-sib families were grown individually in the greenhouse under full light and under low light (40% of ambient) imposed by neutral shade cloth. Low light resulted in a large increase in hypocotyl length and specific leaf area (SLA), a reduction in juvenile biomass and a delayed onset of flowering. Phenotypic selection analysis within each light environment revealed that selection favoured large SLA under low light, but not under high light, suggesting that the observed increase in SLA was adaptive. In contrast, plasticity in the other traits measured was maladaptive (i.e. in the opposite direction to that favoured by selection in the low light environment). We detected significant additive genetic variance in plasticity in most phenotypic traits and in fitness (number of seeds). Using genotypic selection gradient analysis, we found that families with high plasticity in SLA had a lower fitness than families with low plasticity, when the effect of SLA on fitness was statistically kept constant. This indicates that plasticity in SLA incurred a direct fitness cost. However, a cost of plasticity was only expressed under low light, but not under high light. Thus, models on the evolution of phenotypic plasticity will need to incorporate plasticity costs that vary in magnitude depending on environmental conditions.

Intrapopulation sex ratio variation in the salt grass Distichlis spicata.

ABSTRACT In many dioecious plant populations, males and females appear to be spatially segregated, a pattern that is difficult to explain given its potentially high costs. However, in asexually propagating species, spatial segregation of the sexes may be indistinguishable from superficially similar patterns generated by random establishment of a few genets followed by extensive clonal spread and by gender-specific differences in rates of clonal spread. In populations where a significant fraction of individuals are not flowering and gender cannot be assigned to this fraction, apparent spatial segregation of the sexes may be due to differential flowering between the sexes. We confirm reports that flowering ramets of the clonal, perennial grass Distichlis spicata are spatially segregated by sex. We extend these studies in two fundamental ways and demonstrate that this species exhibits true spatial segregation of the sexes. First, using RAPD markers, we estimated that at least 50% of ramets in patches with biased sex ratios represent distinct genotypes. Second, we identified a RAPD marker linked to female phenotype (eliminating the possibility that gender is environmentally determined) and used it to show that the majority of patches exhibit significantly biased sex ratios for both ramets and genets, regardless of flowering status.

POPULATION STRUCTURE ALONG A STEEP ENVIRONMENTAL GRADIENT: CONSEQUENCES OF FLOWERING TIME AND HABITAT VARIATION IN THE SNOW BUTTERCUP, RANUNCULUS ADONEUS.

Few studies have determined how gene flow and selection interact to generate population genetic structure in heterogeneous environments. One way to identify the potential role played by natural selection is to compare patterns of spatial genetic structure between different life cycle stages and among microenvironments. We examined patterns of spatial structure in a population of the snow buttercup (Ranunculus adoneus), using both adult plants and newly emerged seedlings. The study population spans a steep environmental gradient caused by gradual melting of snow within a permanent snowbed. Early-melting sites are characterized by denser vegetation, more fertile soils, and a longer growing season than late-melting sites tens of meters away. The flowering time of R. adoneus is controlled entirely by time of snowmelt, so the contiguous population is phenologically substructured into a series of successively flowering cohorts, reducing the opportunity for direct pollen transfer between early- and late-melting sites. For four highly polymorphic enzyme loci in this tetraploid species, there was subtle, but statistically significant, genetic differentiation between early, middle, and late-melting cohorts; adults usually showed greater differentiation among snowmelt zones than did seedlings. At two loci in adults and one locus in seedlings, homozygotes were more common than predicted at Hardy-Weinberg equilibrium, even when assuming maximum levels of double reduction during meiosis. This pattern suggests the occurrence of self-fertilization and/or population substructure. To determine how spatial isolation and phenological separation each contribute to genetic substructure, we used bivariate regression models to predict the numbers of allele differences between randomly paired individuals as a function of meters separation in space and days separation in flowering time. For newly emerged seedlings, we found that spatial separation was positively associated with genetic difference, but that the additional contribution of phenological separation to genetic difference was not significant. This implies that seeds and/or pollen move effectively across the snowmelt gradient, despite differences in flowering time. As was true for seedlings, spatial separation between paired adults contributed to greater genetic difference, but for a given spatial separation, the genetic difference between adult plants was reduced by phenological separation. This result implies that postemergence selection is favoring at least some seeds that migrate across the snowmelt gradient. Directional gene flow across the snowmelt gradient probably results from a genetic source-sink interaction, that is, the colonization of ecologically marginal late-melting sites by high quality seeds produced by the larger subpopulation in early-melting sites. Effective gene flow from high to low quality microenvironments is likely to impede adaptation to late-melting locations.