Understanding Organisms Using Ecological Observatory Networks.

Human activities are rapidly changing ecosystems around the world. These changes have widespread implications for the preservation of biodiversity, agricultural productivity, prevalence of zoonotic diseases, and sociopolitical conflict. To understand and improve the predictive capacity for these and other biological phenomena, some scientists are now relying on observatory networks, which are often composed of systems of sensors, teams of field researchers, and databases of abiotic and biotic measurements across multiple temporal and spatial scales. One well-known example is NEON, the US-based National Ecological Observatory Network. Although NEON and similar networks have informed studies of population, community, and ecosystem ecology for years, they have been minimally used by organismal biologists. NEON provides organismal biologists, in particular those interested in NEON's focal taxa, with an unprecedented opportunity to study phenomena such as range expansions, disease epidemics, invasive species colonization, macrophysiology, and other biological processes that fundamentally involve organismal variation. Here, we use NEON as an exemplar of the promise of observatory networks for understanding the causes and consequences of morphological, behavioral, molecular, and physiological variation among individual organisms.

Multivariate genetic architecture of the Anolis dewlap reveals both shared and sex-specific features of a sexually dimorphic ornament.

Darwin viewed the ornamentation of females as an indirect consequence of sexual selection on males and the transmission of male phenotypes to females via the 'laws of inheritance'. Although a number of studies have supported this view by demonstrating substantial between-sex genetic covariance for ornament expression, the majority of this work has focused on avian plumage. Moreover, few studies have considered the genetic basis of ornaments from a multivariate perspective, which may be crucial for understanding the evolution of sex differences in general, and of complex ornaments in particular. Here, we provide a multivariate, quantitative-genetic analysis of a sexually dimorphic ornament that has figured prominently in studies of sexual selection: the brightly coloured dewlap of Anolis lizards. Using data from a paternal half-sibling breeding experiment in brown anoles (Anolis sagrei), we show that multiple aspects of dewlap size and colour exhibit significant heritability and a genetic variance-covariance structure (G) that is broadly similar in males (Gm ) and females (Gf ). Whereas sexually monomorphic aspects of the dewlap, such as hue, exhibit significant between-sex genetic correlations (rmf ), sexually dimorphic features, such as area and brightness, exhibit reduced rmf values that do not differ from zero. Using a modified random skewers analysis, we show that the between-sex genetic variance-covariance matrix (B) should not strongly constrain the independent responses of males and females to sexually antagonistic selection. Our microevolutionary analysis is in broad agreement with macroevolutionary perspectives indicating considerable scope for the independent evolution of coloration and ornamentation in males and females.

Intralocus sexual conflict, adaptive sex allocation, and the heritability of fitness.

Intralocus sexual conflict arises when selection favours alternative fitness optima in males and females. Unresolved conflict can create negative between-sex genetic correlations for fitness, such that high-fitness parents produce high-fitness progeny of their same sex, but low-fitness progeny of the opposite sex. This cost of sexual conflict could be mitigated if high-fitness parents bias sex allocation to produce more offspring of their same sex. Previous studies of the brown anole lizard (Anolis sagrei) show that viability selection on body size is sexually antagonistic, favouring large males and smaller females. However, sexual conflict over body size may be partially mitigated by adaptive sex allocation: large males sire more sons than daughters, whereas small males sire more daughters than sons. We explored the evolutionary implications of these phenomena by assessing the additive genetic (co)variance of fitness within and between sexes in a wild population. We measured two components of fitness: viability of adults over the breeding season, and the number of their progeny that survived to sexual maturity, which includes components of parental reproductive success and offspring viability (RS(V) ). Viability of parents was not correlated with adult viability of their sons or daughters. RS(V) was positively correlated between sires and their offspring, but not between dams and their offspring. Neither component of fitness was significantly heritable, and neither exhibited negative between-sex genetic correlations that would indicate unresolved sexual conflict. Rather, our results are more consistent with predictions regarding adaptive sex allocation in that, as the number of sons produced by a sire increased, the adult viability of his male progeny increased.

An experimental test of the role of predators in the maintenance of a genetically based polymorphism.

Polymorphisms provide one of the most useful tools for understanding the maintenance of genetic and phenotypic variation in nature. We have previously described a genetically based polymorphism in dorsal patterning that is expressed by female brown anole lizards, Anolis sagrei, which occur in Bar, Diamond and intermediate Diamond-Bar morphs. Previous studies of island populations in The Bahamas support a role for selection in maintaining the polymorphism, but the agents responsible for this selection remain unclear. We tested two main hypotheses regarding the importance of predation as a selective agent that maintains the polymorphism within populations. First, we tested whether correlational selection favours different combinations of morph, locomotor performance and escape behaviour by measuring morph-specific natural selection on sprint speed, running endurance and the propensity of females to either 'freeze' or 'run' in response to attempted capture. Morphs did not differ in any of these traits, nor did correlational selection consistently favour any particular combinations of morph and antipredator behaviour. Second, we experimentally excluded bird and snake predators from two entire island populations, allowed these predators access to two additional islands and then measured subsequent differences in natural selection on morphs in each population. Predators reduced the survival of Bar and Diamond females, but not of genetically intermediate Diamond-Bar females. These results provide limited evidence that predation may play a role in maintaining this polymorphism, although the functional traits that could account for differential susceptibility to predation remain unclear.

An experimental test for alternative reproductive strategies underlying a female-limited polymorphism.

Polymorphism often corresponds to alternative mating tactics in males, but much less is known about this relationship in females. However, recent work suggests that selection for alternative reproductive strategies in females can maintain genetic variation in important life-history traits. Brown anole lizards (Anolis sagrei) exhibit a genetically based polymorphism in dorsal pattern that is expressed only by females, which occur in bar (B), diamond (D) and intermediate diamond-bar (DB) morphs. Here, we use a combination of natural history data, captive breeding studies and phenotypic manipulations of reproductive investment to test the hypothesis that this polymorphism corresponds to morph-specific patterns of reproductive investment. Three years of data from wild females and two generations of captive breeding revealed no differences among morphs in the frequency of egg production or in the number, frequency, size or sex ratio of offspring. Manipulations of reproductive investment via surgical ovariectomy revealed significant costs of reproduction with respect to survival, growth, immune function and haematocrit, but the magnitudes of these costs did not differ among morphs. Collectively, our results refute the hypothesis that this sex-limited polymorphism is maintained by selection for alternative reproductive strategies. We compare this finding to other systems in which polymorphic females exhibit alternative reproductive tactics and discuss other selective factors that could maintain polymorphism in anoles.

Should hilar lymph nodes be expected in radical nephrectomy specimens?

Lymph node (LN) status is essential in staging both renal cell carcinoma (RCC) and pelvic urothelial carcinoma (PUC). The rate of regional LN involvement is influenced by pathologic tumor stage, extent of the surgical resection, and accuracy of pathologic gross examination. In this study, we assess the presence of hilar LNs in radical nephrectomies (RN) by entirely submitting the hilar fat region (HFR) for microscopic evaluation (ME). Fifty consecutive RNs from 2006 to 2008 were evaluated by a standard gross examination protocol (SGEP) which consisted of palpation and sectioning of the HFR with submission of grossly identified LNs. Subsequently, the entire HFR was re-evaluated and submitted as study's total submission protocol (TSP). The number and disease status of hilar LNs identified by the SGEP and TSP were compared. Fifty RNs (37 clear cell RCC, 6 papillary RCC, 7 PUC) were studied prospectively. Ten of the 50 RNs had LNs identified (20%) with both protocols. Four of the 50 RNs had nodal metastases (8%) with the LN sizes ranging between 1.3 and 2.5 cm (mean 1.8 cm). All nodal metastases were identified by the SGEP. In three RNs (6%), additional minute (mean 0.12 cm) negative LNs not seen by the SGEP were identified by the TSP. LNs are present in only 20% of RNs, even after complete ME of the HFR. The SGEP for identifying hilar LNs in RNs is sufficient for staging and did not lead to underreporting of LN metastases.

Testosterone, growth and the evolution of sexual size dimorphism.

The integration of macroevolutionary pattern with developmental mechanism presents an outstanding challenge for studies of phenotypic evolution. Here, we use a combination of experimental and comparative data to test whether evolutionary shifts in the direction of sexual size dimorphism (SSD) correspond to underlying changes in the endocrine regulation of growth. First, we combine captive breeding studies with mark-recapture data to show that male-biased SSD develops in the brown anole lizard (Anolis sagrei) because males grow significantly faster than females as juveniles and adults. We then use castration surgeries and testosterone implants to show that castration inhibits, and testosterone stimulates, male growth. We conclude by reviewing published testosterone manipulations in other squamate reptiles in the context of evolutionary patterns in SSD. Collectively, these studies reveal that the evolution of SSD has been accompanied by underlying changes in the effect of testosterone on male growth, potentially facilitating the rapid evolution of SSD.

Profiling secondary metabolites of needles of ozone-fumigated white pine (Pinus strobus) clones by thermally assisted hydrolysis/methylation GC/MS.

Plant secondary metabolites have an important role in defense responses against herbivores and pathogens, and as a chemical barrier to elevated levels of harmful air pollutants. This study involves the rapid chemical profiling of phenolic and diterpene resin acids in needles of two (ozone-tolerant and ozone-sensitive) white pine (Pinus strobus) clones, fumigated with different ozone levels (control, and daily events peaking at 80 and 200 ppb) for 40 days. The phenolic and resin acids were measured using thermally assisted hydrolysis and methylation (THM) gas chromatography/mass spectrometry. Short-term fumigation affected the levels of two phenolic acids, i.e., 3-hydroxybenzoic and 3,4-dihydroxybenzoic acids, in that both showed a substantial decrease in concentration with increased ozone dose. The decrease in concentration of these THM products may be caused by inhibition of the plant's shikimate biochemical pathway caused by ozone exposure. The combined occurrence of these two ozone-sensitive indicators has a role in biomonitoring of ozone levels and its impact on forest productivity. In addition, chromatographic profile differences in the major diterpene resin acid components were observed between ozone-tolerant and ozone-sensitive clones. The resin acids anticopalic, 3-oxoanticopalic, 3beta-hydroxyanticopalic, and 3,4-cycloanticopalic acids were present in the ozone-sensitive pine; however, only anticopalic acid was present in the ozone-tolerant clone. This phenotypic variation in resin acid composition may be useful in distinguishing populations that are differentially adapted to air pollutants.

A test of the reproductive cost hypothesis for sexual size dimorphism in Yarrow's spiny lizard Sceloporus jarrovii.

1. Trade-offs between reproduction and growth are central assumptions of life-history theory, but their implications for sexual size dimorphism (SSD) are poorly understood. 2. Adult male Yarrow's spiny lizards Sceloporus jarrovii average 10% larger than adult females. In a low-altitude (1700 m) population, this SSD develops because males grow more quickly than females during the first year of life, particularly during the first female reproductive season. This study tests the hypothesis that SSD develops because female growth is constrained by energetic costs of reproduction. 3. To test for a growth cost of reproduction, I compared growth rates of free-living females that differed, either naturally or experimentally, in reproductive status. Females that naturally delayed reproduction until their second year grew more quickly than females that reproduced as yearlings, and ovariectomized yearlings grew more quickly and to larger sizes than reproductive controls. 4. To determine whether SSD develops in the absence of this inferred reproductive cost, I also studied a high-altitude (2500 m) population in which all females delay reproduction until their second year. Sex differences in growth trajectories were similar to those observed at low altitude, such that males averaged 10% larger than females even prior to female reproduction. 5. Although female growth may be constrained by reproduction, multiple lines of evidence indicate that this cost is insufficient to explain the full magnitude of SSD in S. jarrovii. First, differences in growth of reproductive and nonreproductive females are not observed until the final month of gestation, by which time SSD is already well developed. Second, the growth benefit accruing from experimental inhibition of reproduction accounts for only 32% of the natural sex difference in body size. Finally, SSD develops well in advance of female reproduction in a high-altitude population with delayed maturation.

A computer method for assessing satisfaction with hearing aids.

The purpose of this study was to evaluate a new clinical assessment, the Dynamic Assessment of Hearing Aids (DAHA), for a large clinical population. Unlike traditional questionnaire methods, the DAHA has patients use an intuitive graphical computer interface to record visual analogue ratings of satisfaction with various features of their hearing aids (e.g. clarity, cost, appearance). Data were collected from 191 participants.A subset of participants returned for retest. The DAHA items assess satisfaction with hearing aids within four domains: communication, physical features, sound quality, and personal reactions. The concurrent validity was determined by comparing DAHA results to those obtained with the satisfaction with amplification in daily life (SADL). Ratings for personal reactions to hearing aids indicate the most satisfaction, and ratings for communication (especially group conversations and phone use) indicate the least satisfaction. The DAHA total score was found to have good test/retest and high internal consistency. Concurrent validity was supported by a strong correlation between total scores on the DAHA and the SADL. Results suggest the DAHA maybe an effective tool for clinical use.

Training in neonatal cranial ultrasound: a questionnaire survey.

A questionnaire was sent to every paediatric specialist registrar in the West Midlands to assess the training of paediatric specialist registrars in neonatal cranial ultrasound. 26% had never carried out supervised scans. 51% lacked confidence in performance and 57% in interpretation of scans. The current pattern of training in neonatal cranial ultrasound lacks structure, supervision and assessment of competency.

Effects of simulated thaw on xylem cavitation, residual embolism, spring dieback and shoot growth in yellow birch.

Yellow birch seedlings (Betula alleghaniensis Britt.) that had lost more than 90% of their stem hydraulic conductivity during ambient winter temperatures were exposed to 0 and 20 days of a simulated winter thaw followed by a 48-h freezing treatment at 0, -5, -10, -20 and -30 degrees C. After measuring freezing injury to shoots and roots, the seedlings were placed in a greenhouse where recovery of xylem conductivity and new growth were measured. Shoot xylem cavitation was measured as percent loss of hydraulic conductivity. Shoot freezing injury was assessed by electrolyte leakage (EL) and root freezing injury was assessed by EL and triphenyl tetrazolium chloride reduction. Seedlings pretreated with thaw had higher stem water contents and suffered more freezing damage to roots and shoots (at -20 and -30 degrees C, respectively) than unthawed seedlings. After 3 weeks in a greenhouse, seedlings from the 0, -5 and -10 degrees C freezing treatments showed complete recovery of xylem conductivity, with substantially increased stem water contents. Poor recovery of hydraulic conductivity was observed only in seedlings that were subjected to freezing treatments at -20 and -30 degrees C, regardless of thaw treatment. Of these embolized seedlings, however, only those not previously thawed showed recovery of hydraulic conductivity or regained stem water content after 9 weeks in the greenhouse. Shoot dieback, bud burst and length of new shoots were significantly related to the extent of stem xylem cavitation and freezing injury. We conclude that (1) the simulated winter thaw predisposed yellow birch seedlings to freezing damage in shoots and roots by dehardening tissues and increasing their water content; (2) root freezing damage in turn affected the seedlings' ability to refill embolized stem xylem, resulting in considerable residual xylem embolism after spring refilling; (3) further recovery of stem xylem conductivity was attributable to growth of new vessels; (4) and the permanent residual embolism, together with root and shoot freezing injury, caused increased dieback, bud mortality and reduced growth of new shoots.

Evaluation of a Revised Speech in Noise (RSIN) test.

A revised version of the Speech in Noise (SIN) test was developed by reallocating the recorded test material on the compact disc into different lists (blocks). The goal was to increase the equivalence and reliability of the test blocks to enhance their usefulness in research settings. The Revised Speech in Noise test has four blocks of sentences. Each block comprises twice as many sentences as in the original SIN test. There are also practice sentences for each condition. Forty-two elderly subjects with normal hearing for their age and gender provided data on the equivalence of the new test blocks. The remaining inequalities in mean scores were mostly eliminated using score weighting. Critical differences were developed to promote interpretation of scores from the same individual under different conditions. The revisions substantially improved the equivalence of test blocks and their sensitivity to performance changes. Increased test time is the associated drawback.

Validation of the SADL questionnaire.

OBJECTIVE: To cross-validate the psychometric characteristics of the Satisfaction with Amplification in Daily Life (SADL) questionnaire (Cox & Alexander, 1999), and to explore the SADL's construct validity. DESIGN: Thirteen private practice Audiology clinics each distributed SADL questionnaires, by mail, to 20 adults who had recently obtained hearing aids. The completed questionnaires were returned to a central site and subject anonymity was assured. There were 196 usable responses. RESULTS: Psychometric characteristics of the items were found to be very similar to those reported previously. Thus, the internal validity of the instrument was strongly supported. The assumption that the SADL quantifies satisfaction by assessing its components was evaluated by examining the relationship between SADL scores and scores on a traditional single-item satisfaction measure. A logical and statistically significant relationship was seen between the two measures, thereby supporting the construct validity of both types of data. For private-pay clients, satisfaction scores were very similar to the interim norms published by Cox and Alexander (1999). However, clients whose hearing aids were partly or fully purchased by insurance or benefits programs tended to be more satisfied than interim norms for third-party pay clients derived 5 yr ago. For most types of clients, there was a tendency toward more satisfaction in the Negative Features subscale than observed in our previous research. CONCLUSIONS: Both construct and internal validity of the SADL questionnaire were supported by this research. The previously published interim norms appear to be mostly appropriate for private-pay clients, but might require adjustment in the Negative Features subscale. Further research is needed to explore the relationship between satisfaction and device purchase issues (third-party versus private pay).

Sampling ozone exposure of Canadian forests at different scales: some case studies.

The use of passive samplers in extensive monitoring, such as that used in national forest health monitoring plots, indicates that these devices are able to determine both spatial and temporal differences in ozone exposure of the plots. This allows for categorisation of the plots and the potential for cause-effect analysis of certain forest health responses. Forest exposure along a gradient of air pollution deposition demonstrates large variation in accumulated exposures. The efficacy of using passive samplers for in situ monitoring of forest canopy exposure was also demonstrated. The sampler data produced weak relationships with ozone values from the nearest "continuous" monitor, even though data from colocated samplers showed strong relationships. This spatial variation and the apparent effect of elevation on ozone exposure demonstrate the importance of topography and tree canopy characteristics in plant exposure on a regional scale. In addition, passive sampling may identify the effects of local pollutant gases, such as NO, which may scavenge ozone locally only to increase the production of this secondary pollutant downwind, as atmospheric reactions redress the equilibrium between concentrations of this precursor and those of the generated ozone. The use of passive samplers at the stand level is able to resolve vertical profiles within the stand and edge effects that are important in exposure of understorey and ground flora. Recent case studies using passive samplers to determine forest exposure to ozone indicate a great potential for the development of spatial models on a regional, landscape, and stand level scale.

Verifying loudness perception after hearing aid fitting.

During the verification phase of a hearing aid fitting, clinicians often want to assess the extent to which loudness perceptions for amplified sounds are similar to those typical of normal hearers. This type of verification calls for a criterion for "normal" loudness perception of sounds presented in a sound field. This research sought to answer several questions about the parameters of a valid "normal" criterion for a verification procedure using the Contour test of loudness perception. Loudness data were obtained from 30 listeners with normal hearing. Results indicated that a loudness growth function obtained with earphone listening is not an appropriate normative reference for hearing aid fitting verification. Instead, the normative data should be based on sound field listening. Results also indicated that the same normative function could be used to assess both unilateral and bilateral fittings. Further, it is likely that the same normative function can be used for most frequency responses that are likely to be used in feasible fittings. Finally, it was found that a previously published normative function obtained using an automated test procedure was not faithfully replicated using a carefully executed fully manual test procedure. We concluded that, until a replicable normative function is established, practitioners will need to generate their own local norms to perform postfitting verification of loudness normalization.

Expectations about hearing aids and their relationship to fitting outcome.

Clinicians are often concerned that unrealistic prefitting expectations can have a negative impact on fitting success for new, hearing aid wearers. To investigate this concern and to explore the potential value of measuring expectations, we developed the Expected Consequences of Hearing aid Ownership (ECHO) questionnaire as a companion to the Satisfaction with Amplification in Daily Life questionnaire. Four experiments were conducted to (1) determine realistic expectations for hearing aids, (2) evaluate expectations of new users, (3) measure reliability of prefitting expectations, and (4) assess relationships between prefitting expectations and postfitting satisfaction. Novice hearing aid users were found to have stable prefitting expectations about hearing aids, and these expectations were unrealistically high for the typical individual. There were many different expectation patterns across subjects. Of the four subscales of the ECHO, only one was predictive of the corresponding satisfaction data. Potential clinical applications are described.

Effects of xylem cavitation and freezing injury on dieback of yellow birch (Betula alleghaniensis) in relation to a simulated winter thaw.

Shoot dieback, shoot growth, stem xylem cavitation, stem and root freezing injury, and root pressure were measured in 2-year-old, cold-hardened, potted yellow birch (Betula alleghaniensis Britt.) seedlings that had been subjected to a simulated winter thaw for 0, 5, 10, 19 or 27 days followed by 10 weeks at -10 degrees C. Stem xylem cavitation was determined as percent loss of hydraulic conductivity. Stem freezing injury was measured as electrolyte leakage (EL). Root freezing injury was determined by EL and by triphenyl tetrazolium chloride (TTC) reduction. Thaw duration was significantly correlated with dieback, new shoot growth, stem xylem cavitation, stem and root freezing damage, and root pressure (P < 0.05). In particular, shoot dieback was positively correlated with stem xylem cavitation (P < 0.001), residual stem xylem cavitation (P < 0.01) and root freezing injury (P < 0.010), but only weakly correlated with stem freezing damage (P < 0.05). In roots, freezing damage was negatively correlated with root pressure (P < 0.05), which, in turn, was negatively correlated with residual stem xylem cavitation after root pressure development. In stems, there was no correlation between freezing damage and xylem cavitation. We conclude that long periods of winter thaw followed by freezing resulted in freezing injury to roots concomitant with a reduction in root pressures, leading to poor recovery from freezing-induced xylem embolism.

Measuring Satisfaction with Amplification in Daily Life: the SADL scale.

OBJECTIVE: To develop a self-report inventory to quantify satisfaction with hearing aids. DESIGN: The inventory was developed in several stages. To determine the elements that are most important to satisfaction for most people, we conducted structured interviews and then designed a questionnaire. Hearing aid owners responded to the questionnaire, indicating the relative importance of 14 different elements in their hearing aid satisfaction. Analyses indicated that the elements could be placed into four importance content areas. Trial satisfaction items were designed for each content area and submitted to focus groups to identify highly salient items as well as ambiguous items. A 25-item satisfaction questionnaire then was developed and disseminated to hearing aid owners. Results were obtained from 257 individuals. These data were analyzed to generate the final questionnaire. RESULTS: Fifteen items, divided into four subscales, were selected for the final Satisfaction with Amplification in Daily Life (SADL) questionnaire. The questionnaire yields a Global satisfaction score and a profile of subscale scores, which address Positive Effects, Service and Cost, Negative Features, and Personal Image. A preliminary evaluation of retest stability was conducted with 104 subjects. Ninety percent critical differences for the various scores ranged from 0.9 to 2.0 score intervals on a 7 point scale. CONCLUSIONS: The SADL scale is both brief enough to be clinically acceptable and comprehensive enough to provide a valid assessment of an inherently multidimensional variable. Additional assessment is necessary to refine understanding of its test-retest properties, explore validity issues, and determine clinical, research, and administrative applications of the data.