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Improved knowledge of Dipsacales phylogeny provides a solid framework for studies of character evolution. Although the polarity of many characters can now be confidently established, for others it remains unclear. This results largely from uncertainty about the broader relationships of Dipsacales and is especially problematic for characters that differentiate the two basal lineages, Adoxaceae and Caprifoliaceae. Within Caprifoliaceae, changes in stamen number became decoupled from corolla evolution, a reduction from five to four stamens, coinciding with the origin of the Linnina clade. Subsequently, there was a reduction to two stamens within Morinaceae and to three, two, and one within Valerianaceae. In contrast, within Adoxaceae, stamen number covaries with the number of corolla lobes, although the number of stamens was effectively doubled within Adoxina by the division of each stamen. Fleshy fruits may have evolved separately in Adoxaceae and in Caprifolieae. These vary in the number of seeds and in th...

Progression through the G1/S transition commits cells to synthesize DNA. Cyclin dependent kinase 2 (CDK2) is the major kinase that allows progression through G1/S phase and subsequent replication events. p27 is a CDK inhibitor (CKI) that binds to CDK2 to prevent premature activation of this kinase. Speedy (Spy1), a novel cell cycle regulatory protein, has been found to prematurely activate CDK2 when microinjected into Xenopus oocytes and when expressed in mammalian cells. To determine the mechanism underlying Spy1-induced proliferation in mammalian cell cycle regulation, we used human Spy1 as bait in a yeast two-hybrid screen to identify interacting proteins. One of the proteins isolated was p27; this novel interaction was confirmed both in vitro, using bacterially expressed and in vitro translated proteins, and in vivo, through the examination of endogenous and transfected proteins in mammalian cells. We demonstrate that Spy1 expression can overcome a p27-induced cell cycle arrest to allow for DNA synthesis and CDK2 histone H1 kinase activity. In addition, we utilized p27-null cells to demonstrate that the proliferative effect of Spy1 depends on the presence of endogenous p27. Our data suggest that Spy1 associates with p27 to promote cell cycle progression through the G1/S transition.

1. Commercial reproduction of turkeys relies on pooling of semen from multiple males for inseminations. Understanding how sperm characteristics influence paternity under commercial breeding conditions is important to improving production efficiency. 2. The objective of this study was to evaluate progeny production of individual toms following commercial practices of pooling semen to determine if sperm mobility influences progeny production in field conditions. 3. A total of 104 toms were evaluated for sperm mobility. A subset of 10 toms were housed together and semen was collected, pooled and used to inseminate hens (n = 28). Hens were inseminated at 30 weeks of age and weekly thereafter. 4. Ejaculates from each tom were evaluated on two separate days for sperm mobility. Semen from each tom was diluted and layered upon 6% (wt/vol) Accudenz solution. The sperm suspension was incubated at 41 degrees C for 5 min and absorbance was measured with a spectrophotometer. 5. Toms were ranked by absorbance and categorised as high or low if mobility score was +/- 1 SD from the flock mean (average). 6. For parentage determination, DNA was extracted from tom, hen and poult blood. Poult parentage (n = 276) was determined at one day of age or at 14 weeks by analysis of marker genotypes that were generated by polymerase chain reaction (PCR) amplification of genomic DNA with selected microsatellite markers. 7. Sperm mobility differed across males with absorbance values ranging from 0.147 to 0.366. 8. Findings demonstrate differences in poult production among individual toms when semen from multiple males was pooled and inseminated. Toms classified as high, average and low produced 55, 41 and 4% of the offspring, respectively. 9. It appears that sperm mobility is a trait that influences sperm competition among toms under field conditions where sperm numbers inseminated from individual toms are not controlled or constant and that toms with low sperm mobility produce few offspring.

The objective of this study was to compare a bioassay with a liquid chromatography–fluorescence–mass spectrometryn method for detection of enrofloxacin (ENRO) in incurred eggs. The bioassay developed by our laboratories involves an agar diffusion microbiological method using Klebsiella pneumoniae as an indicator organism. Results demonstrate that both methods are capable of detecting incurred fluoroquinolone residues in eggs. During the 3-day dosing period of hens (Days 1–3) and following drug withdrawal (Days 5, 7, and 9), both of these methods were able to detect incurred ENRO in eggs above the zero tolerance established by the U.S. Food and Drug Administration. The LC–fluorescence–MSn method has the benefit of providing confirmation for fluoroquinolones, while the bioassay may be used as an effective, rapid screening method for detection of fluoroquinolone residues in eggs.

SYNOPSIS. Cladistic biogeographic methods remain susceptible to the confounding effects of ''pseudo-con- gruence'' and ''pseudo-incongruence'' because they were not designed to incorporate information on the absolute timing of the diversification of lineages. Consequently, results from cladistic biogeographic studies are difficult to interpret and cannot be confidently attributed to any particular cause. We illustrate these points with concrete examples, paying special attention to recent work on the biogeography of the Northern Hemisphere, and outline ways in which topological and temporal information might be better integrated. The development of historical biogeography over the last few decades provides general insights into the nature of integration through the life of a discipline. Despite its very promising beginnings, we agree with the assessment of Nelson and Ladiges (1996, p. 54) that cladistic biogeography has yielded few gen- uinely new insights over the last twenty years. When a research program is unproductive for a long period it suggests that something may be wrong with the questions being asked or the methods being used. Here we highlight several elements that have been missing from cladistic biogeographic approaches, focusing par- ticularly on temporal information, and suggest how these might be integrated. The value of incorporating temporal information in such studies has already been recognized (see refer- ences below). Our aim here is to help characterize and illustrate the problem using concrete examples from both the Southern and Northern Hemispheres. The Northern Hemisphere problem provides an especially good example of the difficulties faced by cladistic bio- geographic methods, as well as insights into what can be gained by integrating time and directionality. Al- though we do not propose a new biogeographic meth- od, we highlight several possibilities by comparison to methods recently developed to analyze the coevolution of hosts and their parasites. Finally, in the spirit of the symposium that moti- vated this paper, we offer some general reflections on ''integration.'' Specifically, recent developments in historical biogeography suggest to us that the degree of integration is something that changes in predictable ways through the ontogeny of a discipline, reflecting shifts in the perceived reliability of potentially relevant information.

An efficient multiresidue method for determination of fluoroquinolone antibiotics in eggs has been developed, in which quantitation and confirmation are achieved simultaneously using fluorescence and MS n . With this method, eight fluoroquinolones were analyzed in fortified egg samples at levels of 10–100 ng g −1 . Recoveries for desethylene ciprofloxacin, norfloxacin, ciprofloxacin, danofloxacin, enrofloxacin, orbifloxacin, sarafloxacin and difloxacin were generally in the range 60–100%, with excellent relative standard deviations. The limits of quantitation ranged from 0.1 to 2 ng g −1 in mixed egg. Confirmation was achieved by monitoring the ratios of two prominent product ions in MS 2 (desethylene ciprofloxacin) or MS 3 (all others). The method was found to be applicable to the analysis of egg whites and also egg yolks, with limits of quantitation ranging from 0.1 to 7 ng g −1 (egg white) and 0.1–1.5 ng g −1 (egg yolk). ENRO-incurred mixed eggs, egg whites and egg yolks were also successfully analyzed using this method.

The phylogeny of Rosoideae was investigated using 44 species. Here we report new sequence data from the chloroplast trnL/F region as well as an increased sample of species. The analysis of these new data, along with previously used data from the nuclear ribosomal internal transcribed spacers (ITS), significantly increased resolution as well as confidence for Rosoideae phylogeny. Using both Bayesian inference and parsimony methods, we conducted analyses on the data sets separately and in combination. The resulting phylogenies are congruent with all well‐supported clades of Rosoideae found in previous analyses of ITS or rbcL data. The support for these and other clades is improved, and we consider several clades to be supported well enough to be named. The following clades are given phylogenetic definitions: Sanguisorbeae and its subclades Agrimoniinae and Sanguisorbinae, Potentilleae and its subclades Fragariinae and Potentilla, Roperculina (Rosa + Sanguisorbeae + Potentilleae), and Sanpotina (Sanguisorbea...

The Morinaceae (Dipsacales) contains 13 species placed in Acanthocalyx, Cryptothladia or Morina, and is distributed from the mountains of southeastern Europe through the Himalayas to the Qinghai-Tibetan Plateau, mainly in alpine habitats. Sequence data from two chloroplast regions (the trnK intron and the trnL-F region) and the internal transcribed spacer (ITS) region of nuclear ribosomal DNA were used to infer phylogenetic relationships of Morinaceae and related Dipsacales. Both the nuclear and chloroplast datasets, as well as the combined data, provide strong support for relationships within the Valerina clade, placing Morinaceae as the sister group of a clade containing Valerianaceae and Dipsacaceae plus Triplostegia. The Morinaceae, Acanthocalyx, Cryptothladia, and a clade containing Morina and Cryptothladia, are all supported as monophyletic. However, Morina was found to be paraphyletic in several of our analyses, with Morina longifolia more closely related to Cryptothladia than to other Morina species. There is some evidence that Morina longifolia produces cleistogamous flowers, as do Cryptothladia species. Dispersal-vicariance analyses support the view that Valerina radiated initially within Asia, with subsequent movement into Europe in Morinaceae, Dipsacaceae, and Valerianaceae, and into the New World in Valerianaceae. For Morinaceae, as for a number of plant groups, the Brahmaputra river drainage marks a significant biogeographic divide, although this has been spanned within Acanthocalyx and the Morina-Cryptothladia lineage.

▪ Abstract As better phylogenetic hypotheses become available for many groups of organisms, studies in community ecology can be informed by knowledge of the evolutionary relationships among coexisting species. We note three primary approaches to integrating phylogenetic information into studies of community organization: 1. examining the phylogenetic structure of community assemblages, 2. exploring the phylogenetic basis of community niche structure, and 3. adding a community context to studies of trait evolution and biogeography. We recognize a common pattern of phylogenetic conservatism in ecological character and highlight the challenges of using phylogenies of partial lineages. We also review phylogenetic approaches to three emergent properties of communities: species diversity, relative abundance distributions, and range sizes. Methodological advances in phylogenetic supertree construction, character reconstruction, null models for community assembly and character evolution, and metrics of community phylogenetic structure underlie the recent progress in these areas. We highlight the potential for community ecologists to benefit from phylogenetic knowledge and suggest several avenues for future research.

Two complementary methods for identifying and measuring sulfonamide residues in eggs were developed for use in surveying eggs for potential drug residues. The first method uses liquid chromatography–tandem mass spectrometry (LC–MS–MS) to confirm the presence of sulfonamide residues in eggs. During its validation the limit of confirmation was estimated to be 5–10 ng/g (ppb) depending on the drug. Also, a method for measuring residue level by liquid chromatography with ultraviolet detection (LC–UV) was validated using the same extraction procedure as the confirmatory method. The determinative method was validated over the 50–200 ppb range. Samples were prepared by homogenizing whole egg, extracting with acetonitrile, and cleaning up with a C18 solid-phase extraction cartridge. For confirmation, analytes were separated by gradient LC on a C18 column, ionized by electrospray ionization (ESI), and detected by MS–MS with an ion trap mass spectrometer. For determination, analytes were separated by a different gradient LC procedure and detected by UV at 287 nm. Fifteen drugs were dosed individually in laying hens, and residues of parent drug and/or metabolites were found in eggs for all the drugs. Validation was based on repetitive analyses of control samples, control samples fortified at 100 ppb sulfonamides, and samples of blended incurred eggs.

At the G(2)/M transition of the cell cycle, the cdc25c phosphatase dephosphorylates inhibitory residues of cdc2, and cyclin-B-cdc2 kinase (MPF) is activated. Phosphorylation of cyclin B1 induces its nuclear accumulation, and, since cdc25c is also believed to accumulate and activate shortly before G(2)/M in the nucleus, it has been proposed that this induces cyclin-B1-cdc2 kinase activation. We demonstrate that cyclin B1 phosphorylation has another essential function in vivo: it is required for cdc25c and MPF activation, which does not require nuclear accumulation of cyclin B1, and occurs in the cytoplasm.

Most recent phylogenetic analyses of basal angiosperms have converged on the placement of Amborella as sister to all other extant angiosperms. However, certain recent studies suggest that Amborella and Nymphaeales (water lilies) form a clade sister to all remaining angiosperms or that Nymphaeales alone are the sister to the remaining angiosperms. We report here ( i ) maximum parsimony, maximum likelihood, and Bayesian phylogenetic analyses of 11 genes (>15,000 bp per taxon) for 16 taxa, ( ii ) maximum parsimony analysis for a subset of these genes for 104 taxa, and ( iii ) tests of alternative rootings with the nonparametric bootstrap and the likelihood ratio test with the parametric bootstrap. In addition, we use simulation analyses to examine the amount of bias that may be present in our methods of phylogeny estimation. Amborella continues to receive strong bootstrap support as the sister to all other extant angiosperms, and three of four tests reject alternative hypotheses of the angiosperm root. Although we cannot conclusively choose between Amborella vs. Amborella + Nymphaeales as sister to all other angiosperms, most analyses favor the former rooting.

Fibroblast growth factor receptor 3 (FGFR3) influences a diverse array of biological processes, including cell growth, differentiation, and migration. Activating mutations in FGFR3 are associated with multiple myeloma, cervical carcinoma, and bladder cancer. To identify proteins that interact with FGFR3 and which may mediate FGFR3-dependent signaling, a yeast two-hybrid screen was employed using the cytoplasmic kinase domain of FGFR3 as bait. We identified the adapter protein SH2-B as an FGFR3-interacting protein. Coimmunoprecipitation experiments demonstrate binding of the SH2-Bβ isoform to FGFR3 in 293T cells. Tyrosine phosphorylation of SH2-Bβ was observed when coexpressed with activated FGFR3 mutants such as the weakly activated mutant N540K or the strongly activated mutant K650E, both associated with human developmental syndromes. The extent of tyrosine phosphorylation of SH2-Bβ correlates with receptor activation, suggesting that FGFR3 activation mediates tyrosine phosphorylation of SH2-Bβ. Furthermore, two tyrosine phosphorylation sites of FGFR3, Tyr-724 and Tyr-760, are required for optimal binding of the Src homology-2 (SH2) domain of SH2-Bβ. We also demonstrate the phosphorylation and nuclear translocation of Stat5 by activated FGFR3, which increases in response to overexpression of SH2-Bβ. Taken together, our results identify SH2-Bβ as a novel FGFR3 binding partner that mediates signal transduction.

The decision for a cell to self-replicate requires passage from G1 to S phase of the cell cycle and initiation of another round of DNA replication. This commitment is a critical one that is tightly regulated by many parallel pathways. Significantly, these pathways converge to result in activation of the cyclin-dependent kinase, cdk2. It is, therefore, important to understand all the mechanisms regulating cdk2 to determine the molecular basis of cell progression. Here we report the identification and characterization of a novel cell cycle gene, designated Speedy (Spy1). Spy1 is 40% homologous to the Xenopus cell cycle gene, X-Spy1. Similar to its Xenopus counterpart, human Speedy is able to induce oocyte maturation, suggesting similar biological characteristics. Spy1 mRNA is expressed in several human tissues and immortalized cell lines and is only expressed during the G1/S phase of the cell cycle. Overexpression of Spy1 protein demonstrates that Spy1 is nuclear and results in enhanced cell proliferation. In addition, flow cytometry profiles of these cells demonstrate a reduction in G1 population. Changes in cell cycle regulation can be attributed to the ability of Spy1 to bind to and prematurely activate cdk2 independent of cyclin binding. We demonstrate that Spy1-enhanced cell proliferation is dependent on cdk2 activation. Furthermore, abrogation of Spy1 expression, through the use of siRNA, demonstrates that Spy1 is an essential component of cell proliferation pathways. Hence, human Speedy is a novel cell cycle protein capable of promoting cell proliferation through the premature activation of cdk2 at the G1/S phase transition.

Recent discoveries have dramatically altered traditional views of the stratigraphic distribution and phylogeny of Early Palaeozoic vertebrates and permit a reappraisal of biogeographic patterns and processes over the first 120 million years of vertebrate evolution. Stratigraphic calibration of the phylogenetic trees indicates that most of the pre-Silurian record can be inferred only through ghost ranges. Assessment of the available data suggests that this is due to a shift in ecological niches after the latest Ordovician extinction event and a broadening of geographical range following the amalgamation of Euramerica during the early Silurian. Two major patterns are apparent in the bio- geographic data. Firstly, the majority of jawless fishes with dermoskeletal, plated 'armour' were highly endemic during Cambrian-Ordovician time, with arandaspids restricted to Gondwana, galeaspids to China, and anatolepids, astraspids and, possibly, heterostracans confined to Laurentia. These Laurentian groups began to disperse to other continental blocks as the 'Old Red Sandstone continent' amalgamated through a series of tectonic collisions. The second maj or pattern, in contrast, encompasses a number of microsquamous and naked, jawed and jawless primitive vertebrates such as conodonts, thelodonts, placo- derms, chondrichthyans and acanthodians, which dispersed rapidly and crossed oceanic barriers to attain cosmopolitan distributions, although many have Laurentian origins. A clear difference in dispersal potential exists between these two types of fishes. Overall, the development of biogeographic patterns in Early Palaeozoic vertebrates involved a complex interaction of dispersal, vicariance and tectonic convergence.

It has been contended that Reif's odontode regulation theory is a rival and alternative to Stensioand Orvig's lepidomorial theory as means of explaining the evolution of development of the vertebrate dermal and oral skeleton. The lepidomorial theory is a pattern-based theory that provides a homological framework that goes further than the odontode regulation theory in comparing dental papillae and their products, and it provides an explanatory mechanism for such relationships a pos- teriori. In contrast, the odontode regulation theory is process-based and observes only developmental similarity, providing no means of identifying homologies beyond this. The lepidomorial theory is superior to the odontode regulation theory in its ability to trace homology through the evolution of development of the dermal and oral skeleton. The criteria proposed to identify homology between scales—either within a given individual or taxon, or between different individuals or taxa—are, pri- marily, vascular architecture and, secondarily, external morphology. External morphology may be excluded on Reif's argument for the overarching principle of differentiation, a hypothesis supported by recent advances in the understanding of dental morphogenesis. Vascular architecture is potentially useful but appears to be determined by tooth/scale morphology rather than reflecting historical (phy- logenetic) constraint. Data on the development of epithelial appendages, including teeth, scales, and feathers, indicate that individual primordia develop through progressive differentiation of originally larger, homogenous morphogenetic fields. Thus, there is no mechanism of ontogenetic developmental concrescence, just differentiation. Phylogenetic patterns of concrescence and differentiation are sim- ilarly achieved through ontogenetic developmental differentiation, or a lack thereof. In practice, how- ever, it is not possible to distinguish between patterns of phylogenetic concrescence and differenti- ation because there is no means of identifying homology between individual elements within a squa- mation, or a dentition (in almost all instances). Thus, phylogenetic patterns of increase and decrease in the numbers of elements constituting dentitions or dermal elements are best described as such; further attempts to constrain precise underlying patterns remain without constraint and outside the realms of scientific enquiry. The application of the homology concept in the dermal and visceral skel- etons is explored and it is determined that odontodes are serial homologs, conform only to the bio- logical homology concept at this level of observation, and are devoid of phylogenetic meaning. It is concluded that Reif's theory is close to a universal theory of the evolution of development for the dermoskeleton and dentition, and additional components of theory, including the regulatory basis of temporal and spatial patterning, are tested and extended in light of data on the development of the chick feather array. Finally, the dermoskeleton is identified as an exemplary system for examining the regulatory basis of patterning and morphogenesis as it encompasses and surpasses the repertoire of established model organ systems.

Whole genome duplication (WGD) has occurred in many lineages within the tree of life and is invariably invoked as causal to evolutionary innovation, increased diversity, and extinction resistance. Testing such hypotheses is problematic, not least since the timing of WGD events has proven hard to constrain. Here we show that WGD events can be dated through molecular clock analysis of concatenated gene families, calibrated using fossil evidence for the ages of species divergences that bracket WGD events. We apply this approach to dating the two major genome duplication events shared by all seed plants ( ζ ) and flowering plants ( ɛ ), estimating the seed plant WGD event at 399–381 Ma, and the angiosperm WGD event at 319–297 Ma. These events thus took place early in the stem of both lineages, precluding hypotheses of WGD conferring extinction resistance, driving dramatic increases in innovation and diversity, but corroborating and qualifying the more permissive hypothesis of a ‘lag-time’ in realizing the effects of WGD in plant evolution.

Sleep disturbances, depression, and inactivity are common non-motor problems in Parkinson’s disease (PD). The effects of Whole Body Periodic Acceleration (WBPA) in persons with PD on sleep regulation and activity levels have yet to be evaluated.The objective was to assess whether WBPA improves sleep and activity in persons with PD over 4 weeks compared to gender matched controls.Participants included 5 PD and 5 controls (8 men; 2 women), age 74.10 ± 9.25. Activity trackers monitored sleep, awakenings, and step counts over 24 hours for 6 weeks. Baseline measurements included sleep 30 days prior to study, depression (PHQ-9), and Quality of Life (PDQ-8). WBPA duration was 45 minutes, 3 times per week for 4 weeks. A 2 by 6 repeated measure and paired t tests were used to assess data. A significant decrease was found in BP pre vs. post p .019; PD group had improved depression p .025. Minimally Clinically Significant Differences (MCID) were also found in the PD participants, the PSQI (4.25) points. Self-reports for WBPA displayed improvements in depression and sleep. Decreased physical activity in those with mild to moderate PD was evidenced. WBPA may have a significant role in PD management of non-motor symptoms. Further study with larger groups is warranted using WBPA.

Pond populations in Cape Cod described as Persicaria puritanorum ( ≡ Polygonum puritanorum ) have been treated by recent authors as a synonym of P. maculosa ( ≡ Polygonum persicaria ) based on gross morphology. However, consistent differences in leaf and inflorescence features raise the question as to whether this is an extreme phenotype of P. maculosa , an ecotype of that species, or a distinct taxon. In this study, we analyzed nuclear ITS region and chloroplast mat K region DNA sequences from 37 populations of 11 species within section Eupersicaria and counted the chromosome number of individuals from the pond populations in question. We also compared leaf size and shape of plants from these populations with those of the typical P. maculosa plants by measuring selected herbarium specimens. Phylogenetic trees inferred from sequence data indicate a separation of pond populations from P. maculosa suggesting to recover P. puritanorum as a distinct species. Chromosome counts of pond population plants (2n = 60) strongly support this result, and furthermore suggest the possibility of a hybrid polyploid origin of P. puritanorum . A leaf morphological trait (greater relative height of the maximum width point from the leaf base) also supports the separation of pond populations from P. maculosa . P. punctata allied to P. hydropiper in the ITS-based tree, and to pond populations as well as P. hydropiperoides and P. opelousana in the matK -based tree, suggesting the possibility of hybrid origin of P. punctata . Our ITS sequence analyses also reveal distinct two lineages in P. lapathifolia but the chloroplast matK region sequences of P. pensylvanica and two lineages of P. lapathifolia are almost identical suggesting cytoplasmic introgression or allopolyploid speciation sharing the maternal lineage.

The evolution of ecological idiosyncrasies in Madagascar has often been attributed to selective pressures stemming from extreme unpredictability in climate and resource availability compared to other tropical areas. With the exception of rainfall, few studies have investigated these assumptions. To assess the hypothesis that Madagascar's paucity of frugivores is due to unreliability in fruiting resources, we use statistical modeling to analyze phenology datasets and their environmental correlates from two tropical wet forests, the Réserve Naturelle Intégrale Betampona in Madagascar, and Kibale National Park in Uganda. At each site we found that temperature is a good environmental predictor of fruit availability. We found no evidence of a significant difference in the predictability of fruit availability between the two sites, although the shorter duration of phenological monitoring at Betampona (two years, versus 15 years at Kibale) limits our ability to infer long-term patterns. Comparisons of long-term temperature data from each site (15 years from Kibale and 14 from Betampona) indicate that temperature is more predictable at Betampona than at Kibale. However, there does appear to be a difference between the two sites in the total fruit availability at any given time, with fruit being generally less abundant at Betampona. Our results appear contrary to the prevailing hypothesis of a selective force imposed by unpredictable resource availability or temperature, and we suggest other possible explanations for Madagascar's unique biota.

Integration influences patterns of trait evolution, but the relationship between these patterns and the degree of trait integration is not well understood. To explore this further, we study a specialized pollination mechanism in conifers whose traits are linked through function but not development. This mechanism depends on interactions among three characters: pollen that is buoyant, ovules that face downward at pollination, and the production of a liquid droplet that buoyant grains float through to enter the ovule. We use a well-sampled phylogeny of conifers to test correlated evolution among these characters and specific sequences of character change. Using likelihood models of character evolution, we find that pollen morphology and ovule characters evolve in a concerted manner, where the flotation mechanism breaks down irreversibly following changes in orientation or drop production. The breakdown of this functional constraint, which may be facilitated by the lack of developmental integration among the constituent traits, is associated with increased trait variation and more diverse pollination strategies. Although this functional "release" increases diversity in some ways, the irreversible way in which the flotation mechanism is lost may eventually result in its complete disappearance from seed plant reproductive biology.

The role of teeth and jaws, as innovations that underpinned the evolutionary success of living jawed vertebrates, is well understood, but their evolutionary origins are less clear. The origin of teeth, in particular, is mired in controversy with competing hypotheses advocating their origin in external dermal denticles ("outside-in") versus a de novo independent origin ("inside-out"). No evidence has ever been presented demonstrating materially the traditional "outside-in" theory of teeth evolving from dermal denticles, besides circumstantial evidence of a commonality of structure and organogenesis, and phylogenetic evidence that dermal denticles appear earlier in vertebrate phylogeny that do teeth. Meanwhile, evidence has mounted in support of "inside-out" theory, through developmental studies that have indicated that endoderm is required for tooth development, and fossil studies that have shown that tooth-like structures evolved before dermal denticles (conodont dental elements), that tooth replacement evolving before teeth (thelodont pharyngeal denticles), and that teeth evolved many times independently through co-option of such structures. However, the foundations of "inside-out" theory have been undermined fatally by critical reanalysis of the evidence on which it was based. Specifically, it has been shown that teeth develop from dermal, endodermal or mixed epithelia and, therefore, developmental distinctions between teeth and dermal denticles are diminished. Furthermore the odontode-like structure of conodont elements has been shown to have evolved independently of dermal and internal odontodes. The tooth-like replacement encountered in thelodont pharyngeal odontodes has been shown to have evolved independently of teeth and tooth replacement and teeth have been shown to have evolved late within the gnathostome stem lineage indicating that it is probable, if not definitive, that teeth evolved just once in gnathostome evolution. Thus, the "inside-out" hypothesis must be rejected. The phylogenetic distribution of teeth and dermal denticles shows that these odontodes were expressed first in the dermal skeleton, but their topological distribution extended internally in association with oral, nasal and pharyngeal orifices, in a number of distinct evolutionary lineages. This suggests that teeth and oral and pharyngeal denticles emerged phylogenetically through extension of odontogenic competence from the external dermis to internal epithelia. Ultimately, internal and external odontodes appear to be distinct developmental modules in living jawed vertebrates, however, the evidence suggests that this distinction was not established until the evolution of jawed vertebrates, not merely gnathostomes.

• Premise of the study: Despite recent progress, significant uncertainties remain concerning relationships among early-branching lineages within Viburnum (Adoxaceae), prohibiting a new classification and hindering studies of character evolution and the increasing use of Viburnum in addressing a wide range of ecological and evolutionary questions. We hoped to resolve these issues by sequencing whole plastid genomes for representative species and combining these with molecular data previously obtained from an expanded taxon sample.• Methods: We performed paired-end Illumina sequencing of plastid genomes of 22 Viburnum species and combined these data with a 10-gene data set to infer phylogenetic relationships for 113 species. We used the results to devise a comprehensive phylogenetic classification and to analyze the evolution of eight morphological characters that vary among early-branching lineages.• Key results: With greatly increased levels of confidence in most of the early branches, we propose a phylogenetic classification of Viburnum, providing formal phylogenetic definitions for 30 clades, including 13 with names recognized under the International Code of Nomenclature for Algae, Fungi, and Plants, eight with previously proposed informal names, and nine newly proposed names for major branches. Our parsimony reconstructions of bud structure, leaf margins, inflorescence form, ruminate endosperm, extrafloral nectaries, glandular trichomes, palisade anatomy, and pollen exine showed varying levels of homoplasy, but collectively provided morphological support for some, though not all, of the major clades.• Conclusions: Our study demonstrates the value of next-generation plastid sequencing, the ease of creating a formal phylogenetic classification, and the utility of such a system in describing patterns of character evolution.

Purpose. The use of benzodiazepines remains a source of controversy. Some prescribers believe that they are beneficial and espouse their use; others regard their risk:benefit ratio as too adverse for any but occasional use. This review considers these viewpoints based on the appropriate literature. Survey. The recent English-language literature relating to this topic was surveyed. The publications proved too heterogeneous for a formal meta-analysis, so a descriptive review is provided. Overview. Surveys of benzodiazepine use provide data mainly from the UK, Europe and North America. Prescribing patterns varied widely but long-term usage is common and sometimes the norm. Conclusions. Long-term prescription of benzodiazepines still takes place despite general warnings from the medical and other professions and drug regulatory bodies that long-term use is unjustified both from the lack of a systemic database establishing such efficacy and a large literature documenting the risks of long-term usage, such as dependence. The young and the old are particularly at risk. Continued monitoring is essential, but the regulatory authorities may need to take a more active role in curbing such undesirable practice.

Many major branches in the Tree of Life are marked by stereotyped body plans that have been maintained over long periods of time. One possible explanation for this stasis is that there are genetic or developmental constraints that restrict the origin of novel body plans. An alternative is that basic body plans are potentially quite labile, but are actively maintained by natural selection. We present evidence that the conserved floral morphology of a species-rich flowering plant clade, Malpighiaceae, has been actively maintained for tens of millions of years via stabilizing selection imposed by their specialist New World oil-bee pollinators. Nine clades that have lost their primary oil-bee pollinators show major evolutionary shifts in specific floral traits associated with oil-bee pollination, demonstrating that developmental constraint is not the primary cause of morphological stasis in Malpighiaceae. Interestingly, Malpighiaceae show a burst in species diversification coinciding with the origin of this plant–pollinator mutualism. One hypothesis to account for radiation despite morphological stasis is that although selection on pollinator efficiency explains the origin of this unique and conserved floral morphology, tight pollinator specificity subsequently permitted greatly enhanced diversification in this system.

The process by which excitatory neurons are generated and mature during the development of the cerebral cortex occurs in a stereotyped manner; coordinated neuronal birth, migration, and differentiation during embryonic and early postnatal life are prerequisites for selective synaptic connections that mediate meaningful neurotransmission in maturity. Normal cortical function depends upon the proper elaboration of neurons, including the initial extension of cellular processes that lead to the formation of axons and dendrites and the subsequent maturation of synapses. Here, we examine the role of cell-based signaling via the receptor tyrosine kinase EphA7 in guiding the extension and maturation of cortical dendrites. EphA7, localized to dendritic shafts and spines of pyramidal cells, is uniquely expressed during cortical neuronal development. On patterned substrates, EphA7 signaling restricts dendritic extent, with Src and Tsc1 serving as downstream mediators. Perturbation of EphA7 signaling in vitro and in vivo alters dendritic elaboration: Dendrites are longer and more complex when EphA7 is absent and are shorter and simpler when EphA7 is ectopically expressed. Later in neuronal maturation, EphA7 influences protrusions from dendritic shafts and the assembling of synaptic components. Indeed, synaptic function relies on EphA7; the electrophysiological maturation of pyramidal neurons is delayed in cultures lacking EphA7, indicating that EphA7 enhances synaptic function. These results provide evidence of roles for Eph signaling, first in limiting the elaboration of cortical neuronal dendrites and then in coordinating the maturation and function of synapses.

O'Leary et al. (O'Leary et al. 2013 Science 339, 662–667. (doi:10.1126/science.1229237)) performed a fossil-only dating analysis of mammals, concluding that the ancestor of placentals post-dated the Cretaceous–Palaeogene boundary, contradicting previous palaeontological and molecular studies that placed the ancestor in the Cretaceous. They incorrectly used fossil ages as species divergence times for crown groups, while in fact the former should merely form minimum-age bounds for the latter. Statistical analyses of the fossil record have shown that crown groups are significantly older than the oldest ingroup fossil, so that fossils do not directly reflect the true ages of clades. Here, we analyse a 20 million nucleotide genome-scale alignment in conjunction with a probabilistic interpretation of the fossil ages from O'Leary et al. Our combined analysis of fossils and molecules demonstrates that Placentalia originated in the Cretaceous.

The fossil faunas of the Cambrian provide the only direct insight into the assembly of animal body plans. However, for many animal groups, their early fossil record is linked to disarticulated remains, interpretation of which is problematic since they possess few characters from which their affinity to phyla can be established and, indeed, few characters at all. One such group is the tommotiids, which has been interpreted, on the basis of skeletal anatomy, as a paraphyletic assemblage uniting brachiopods and phoronids, through the acquisition and subsequent modification, or loss, of an imbricated set of dorsal phosphatic sclerites. Here we present a reexamination of the fossil evidence uniting the tommotiids and brachiopods, supplemented with new anatomical data from synchrotron radiation X-ray tomographic microscopy of key tommotiid taxa. The characters used to support the complex hypothesis of character evolution in the brachiopod stem lineage relies on scleritome reconstructions and inferred mode of life which themselves rely on brachiopods being chosen as the interpretative model. We advocate a more conservative interpretation of the affinity of these fossils, based a priori on their intrinsic properties, rather than the modern analogue in whose light they have been interpreted.

Studies of the development of organisms can reveal crucial information on homology of structures. Developmental data are not peculiar to living organisms, and they are routinely preserved in the mineralized tissues that comprise the vertebrate skeleton, allowing us to obtain direct insight into the developmental evolution of this most formative of vertebrate innovations. The pattern of developmental processes is recorded in fossils as successive stages inferred from the gross morphology of multiple specimens and, more reliably and routinely, through the ontogenetic stages of development seen in the skeletal histology of individuals. Traditional techniques are destructive and restricted to a 2-D plane with the third dimension inferred. Effective non-invasive methods of visualizing paleohistology to reconstruct developmental stages of the skeleton are necessary. In a brief survey of paleohistological techniques we discuss the pros and cons of these methods. The use of tomographic methods to reconstruct development of organs is exemplified by the study of the placoderm dentition. Testing evidence for the presence of teeth in placoderms, the first jawed vertebrates, we compare the methods that have been used. These include inferring development from morphology, and using serial sectioning, microCT or synchrotron X-ray tomographic microscopy (SRXTM), to reconstruct growth stages and directions of growth. The ensuing developmental interpretations are biased by the methods and degree of inference. The most direct and reliable method is using SRXTM data to trace sclerochronology. The resulting developmental data can be used to resolve homology and test hypotheses on the origin of evolutionary novelties., Journal of paleontology, 88 (4), ISSN:0022-3360, ISSN:1937-2337

The article discusses the significant role of biodiversity in the evolution. Topics discussed include importance of applying evolutionary principles in conservation and human well-being, fundamental relevance of evolution in the production, maintenance, and future of biodiversity and biodiversity arises from the approach that treats groups of taxa with similar traits as a significant functional unit.

Cancer is a major public health problem worldwide. In the United States alone, 1 in 4 deaths is due to cancer and for 2013 a total of 1,660,290 new cancer cases and 580,350 cancer-related deaths are projected. Comprehensive profiling of multiple cancer genomes has revealed a highly complex genetic landscape in which a large number of altered genes, varying from tumor to tumor, impact core biological pathways and processes. This has implications for therapeutic targeting of signaling networks in the development of treatments for specific cancers. The NFκB transcription factor is constitutively active in a number of hematologic and solid tumors, and many signaling pathways implicated in cancer are likely connected to NFκB activation. A critical mediator of NFκB activity is TGFβ-activated kinase 1 (TAK1). Here, we identify TAK1 as a novel interacting protein and target of fibroblast growth factor receptor 3 (FGFR3) tyrosine kinase activity. We further demonstrate that activating mutations in FGFR3 associated with both multiple myeloma and bladder cancer can modulate expression of genes that regulate NFκB signaling, and promote both NFκB transcriptional activity and cell adhesion in a manner dependent on TAK1 expression in both cancer cell types. Our findings suggest TAK1 as a potential therapeutic target for FGFR3-associated cancers, and other malignancies in which TAK1 contributes to constitutive NFκB activation. © 2014 Salazar et al.

Drug and pesticide residues in edible poultry tissues and eggs are a human food safety concern. Quantifying and predicting the concentrations of residues in eggs is a particular challenge because of the unique aspects of egg development and compartmentalisation of the yolk and white. For a number of years, research efforts have been directed at examining residue incorporation nto eggs and developing models to predict residue incursion. This paper will review the physiology of egg yolk formation as it relates to residue incursion, the models developed to understand and quantify residue patterns, and the use of magnetic resonance imaging to detect residue incorporation into eggs.

Geological and climatological processes that have impacted the biota of the Northern Hemisphere during the Tertiary are expected to yield little resolution when area cladograms are compared without taking the timing of diversification into account. In an attempt to establish a set of appropriate phylogenetic comparisons, we distinguished between a Pacific track involving (minimally) China, Japan, and eastern North America but not Europe, and an Atlantic track involving China, Europe, and eastern North America but not Japan (or, in most cases, western North America). Within the two Atlantic‐track taxa considered here—Liquidambar and Cercis—European and North American species are more closely related to one another than they are to the Asian species. Within a set of five Pacific‐track taxa—Hamamelis, Weigela‐Diervilla, Triosteum, Buckleya, and Torreya—we see all possible relationships involving China, Japan, and eastern North America. Estimates of minimum divergence times between Old World and New World lin...

Conodonts were mostly small, elongate, eel-shaped marine animals that inhabited a variety of environments in Paleozoic and Triassic seas. Although long enigmatic, conodonts are now regarded as vertebrates and their closely controlled fossil record is not only the most extensive of all vertebrates, but it also makes conodonts the fossils of choice in upper Cambrian through Triassic biostratigraphy. Conodonts were soft-bodied except for a variety of phosphatic elements that formed a distinctive feeding apparatus. Post-mortal dissociation of the apparatus and subsequent jumbling of its elements on the sea floor led, from 1856 to about 1966, to development of an artificial, form-based taxonomy that was utilitarian, but clearly unsatisfactory as a vehicle for understanding the group in biologic terms.Natural assemblagesof elements, discovered between 1879 and 1952, have been interpreted as undisturbed skeletal apparatuses, and in the mid-1960s it was determined that original composition of the apparatuses of many species could be reconstructed and statistically evaluated from collections of disjunct elements by various grouping procedures. These determinations led to an emphasis onmultielement taxonomyby most (but not all) students of conodonts. Even so, only about a third of the approximately 550 valid conodont genera, have been established (or re-interpreted) in multielement terms and this makes any of the several extant schemes of suprageneric classification phylogenetically suspect. We comment on a recent scheme that recognizes 41 families assigned to some 7 orders, and suggest how it might be modified so as to square with principles of phylogenetic systematics.

The Malpighiaceae are a family of ;1250 species of predominantly New World tropical flowering plants. Infrafamilial classification has long been based on fruit characters. Phylogenetic analyses of chloroplast DNA nucleotide sequences were analyzed to help resolve the phylogeny of Malpighiaceae. A total of 79 species, representing 58 of the 65 currently recognized genera, were studied. The 39 region of the gene ndhF was sequenced for 77 species and the noncoding intergenic spacer region trnL-F was sequenced for 65 species; both sequences were obtained for the outgroup, Humiria (Humiriaceae). Phylogenetic relationships inferred from these data sets are largely congruent with one another and with results from combined analyses. The family is divided into two major clades, recognized here as the subfamilies Byrsonimoideae (New World only) and Malpighioideae (New World and Old World). Niedenzu’s tribes are all polyphyletic, suggesting extensive convergence on similar fruit types; only de Jussieu’s tribe Gaudichaudieae and Anderson’s tribes Acmanthereae and Galphimieae are monophyletic. Fleshy fruits evolved three times in the family and bristly fruits at least three times. Among the wing-fruited vines, which constitute more than half the diversity in the family, genera with dorsal-winged samaras are fairly well resolved, while the resolution of taxa with lateral-winged samaras is poor. The trees suggest a shift from radially symmetrical pollen arrangement to globally symmetrical pollen at the base of one of the clades within the Malpighioideae. The Old World taxa fall into at least six and as many as nine clades.

Direct sample introduction (DSI) or "dirty sample injection" is a rapid, rugged, and inexpensive approach to large volume injection in gas chromatography (GC) for semivolatile analytes such as pesticides. DSI of complex samples such as eggs requires a very selective detection technique, such as tandem mass spectrometry (MS-MS), to determine the analytes among the many semivolatile matrix components that also appear. In DSI, the nonvolatile matrix components that normally would contaminate the GC system in traditional injection methods remain in a disposable microvial, which is removed after every injection. For example, 3 microg of nonvolatile residue typically remained in the microvial after an injection of egg extract using the DSI method. This analytical procedure involves the following: (i) weighing 10 g of egg in a centrifuge tube and adding 2 g of NaCl and 19.3 mL of acetonitrile (MeCN); (ii) blending for 1 min using a probe blender; (iii) centrifuging for 10 min; and (iv) analyzing 10 microL (5 mg of egg equivalent) of the extract using DSI/GC/MS-MS. No sample cleanup or solvent evaporation steps were required to achieve quantitative and confirmatory results with

BackgroundAlthough the efficacy of antidepressants has been demonstrated in randomised, controlled clinical trials, it is how an antidepressant is used in clinical practice that determines its clinical effectiveness, or real-world efficacy.AimsTo explore the frequency with which antidepressants are used at adequate dose and duration to obtain remission of symptoms and prevent relapse in clinical practice and discuss potential implications for clinical outcomes.MethodStudies of antidepressant prescribing were reviewed and comparisons made between antidepressant classes and individual compounds within those classes.ResultsNaturalistic studies show that patients who begin therapy on tricyclic antidepressants often receive sub-therapeutic doses for inadequate duration; conversely patients who begin therapy on selective serotonin reuptake inhibitors more often receive an adequate dose of therapy for a longer duration.ConclusionsHow antidepressants are used in clinical practice can determine the clinical outcomes that are achieved. Antidepressants that are more forgiving of sub-optimal prescribing and use patterns by providers and patients, respectively, may help to improve real-world efficacy.

Fibroblast growth factor receptor 3 (FGFR3) mutations are frequently involved in human developmental disorders and cancer. Activation of FGFR3, through mutation or ligand stimulation, results in autophosphorylation of multiple tyrosine residues within the intracellular domain. To assess the importance of the six conserved tyrosine residues within the intracellular domain of FGFR3 for signaling, derivatives were constructed containing an N-terminal myristylation signal for plasma membrane localization and a point mutation (K650E) that confers constitutive kinase activation. A derivative containing all conserved tyrosine residues stimulates cellular transformation and activation of several FGFR3 signaling pathways. Substitution of all nonactivation loop tyrosine residues with phenylalanine rendered this FGFR3 construct inactive, despite the presence of the activating K650E mutation. Addition of a single tyrosine residue, Y724, restored its ability to stimulate cellular transformation, phosphatidylinositol 3-kinase activation, and phosphorylation of Shp2, MAPK, Stat1, and Stat3. These results demonstrate a critical role for Y724 in the activation of multiple signaling pathways by constitutively activated mutants of FGFR3.

Little is known regarding the internal anatomy of thelodonts, with most of the available information coming from a single specimen, the holotype of Turinia pagei (Powrie). Previous descriptions have led to many partial, or conflicting interpretations. Herein, we describe fully the anatomy of T. pagei based on the holotype and additional material. T. pagei possessed a branchial system composed of eight pairs of gills, a buccal/nasohypophyseal region lined with minute denticles, comparable to buccopharyngeal denticles of sharks, and possessed a stomach which is preserved by sediment infill in the holotype specimen. Contrast between the petrological character of the gut infill and the sediment in which the animal is preserved suggests that the gut was infilled in vivo and that T. pagei was probably a deposit feeder. Phylogenetic analysis resolves T. pagei and the Galeaspida as sister-taxa, comprising a sister-group to the Osteostraci plus jawed vertebrates. In contrast to the view that has prevailed hitherto, thelodonts with a dorsoventrally compressed cross-sectional profile comprise a monophyletic group, of which T. pagei is the least derived member. The furcacaudiforms are resolved as an unnatural group, one taxon being the sister taxon to the ‘conventional’ thelodont clade, and the other, the sister taxon to this clade plus galeaspids, osteostracans and jawed vertebrates. The analysis agrees with the earlier view that thelodonts lack distinct synapomorphies, but we argue that distinct synapomorphies are not a requisite of monophyly.

A numerical cladistic analysis of the conodont family Palmatolepidae has been undertaken to determine the applicability of the technique to group-wide systematic revision. Results suggest a new hypothesis of relationships that is considerably more parsimonious than trees compatible with existing hypotheses of relationships, or trees that are even loosely constrained stratigraphically. This may occur either because the fossil record is incomplete, because taxon sampling for the cladistic analysis is low, or because the most parsimonious trees approximate the true tree less well than do stratigraphically-constrained trees (or because of a combination of these factors). Although more taxa and more characters would be preferable in choosing between these possibilities, the tree derived solely from morphological data is adopted. Thus, stratigraphic data can be used to test hypotheses of relationships and construct phylogenies; hypotheses of relationships can be used to test the completeness of the conodont fossil record. Existing schemes of classification within the Palmatolepidae are rejected because most groups within them are either polyphyletic or paraphyletic. A new scheme is presented. Character changes suggest correlated, progressive and mosaic evolution within the Palmatolepidae. Parsimony analysis of partitioned datasets indicates that more phylogenetic information can be recovered from S rather than P or M element positions, although data from all three positional groups are preferable to data from just one. Thus, multielement taxonomy is essential to the resolution of conodont interrelationships.

The visual cortex in primates is parcellated into cytoarchitectonically, physiologically, and connectionally distinct areas: the striate cortex (V1) and the extrastriate cortex, consisting of V2 and numerous higher association areas [1]. The innervation of distinct visual cortical areas by the thalamus is especially segregated in primates, such that the lateral geniculate (LG) nucleus specifically innervates striate cortex, whereas pulvinar projections are confined to extrastriate cortex [2–8]. The molecular bases for the parcellation of the visual cortex and thalamus, as well as the establishment of reciprocal connections between distinct compartments within these two structures, are largely unknown. Here, we show that prospective visual cortical areas and corresponding thalamic nuclei in the embryonic rhesus monkey ( Macaca mulatta ) can be defined by combinatorial expression of genes encoding Eph receptor tyrosine kinases and their ligands, the ephrins, prior to obvious cytoarchitectonic differentiation within the cortical plate and before the establishment of reciprocal connections between the cortical plate and thalamus. These results indicate that molecular patterns of presumptive visual compartments in both the cortex and thalamus can form independently of one another and suggest a role for EphA family members in both compartment formation and axon guidance within the visual thalamocortical system.

A multiresidue solid phase extraction (SPE) method for the isolation and subsequent gas chromatographic determination of nonpolar organochlorine and polar organophosphorus pesticide residues in eggs is described. The method uses an acetonitrile extraction followed by an SPE cleanup using graphitized carbon black and aminopropyl SPE columns. Organophosphorus pesticides are determined by gas chromatography with flame photometric detection. After further cleanup of the extract using Florisil SPE columns, organochlorine pesticides are determined by gas chromatography with electron capture detection. Studies were performed using eggs containing both fortified and incurred pesticide residues. The average recoveries were 86-108% for 8 fortified organochlorine pesticide residues and 61-149% for 28 fortified organophosphorus pesticide residues.

We have extended our studies of angiosperm phylogeny based on a pair of duplicated phytochrome (PHY) genes, PHYA and PHYC. Phylogenetic analyses of sequences from 52 species yield unrooted gene networks in which all of the elements resolved in our previous analysis of 26 species appear. Amborella still emerges as the sister group of all other angiosperms. However, we cannot reject alternative rootings in which water lilies, either alone or in combination with Amborella, are basal. \documentclass{aastex} \usepackage{amsbsy} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{bm} \usepackage{mathrsfs} \usepackage{pifont} \usepackage{stmaryrd} \usepackage{textcomp} \usepackage{portland,xspace} \usepackage{amsmath,amsxtra} \usepackage[OT2,OT1]{fontenc} \newcommand\cyr{ \renewcommand\rmdefault{wncyr} \renewcommand\sfdefault{wncyss} \renewcommand\encodingdefault{OT2} \normalfont \selectfont} \DeclareTextFontCommand{\textcyr}{\cyr} \pagestyle{empty} \DeclareMathSizes{10}{9}{7}{6} \begin{document} \landscape $...

A multiresidue method was developed for the determination of fluoroquinolones in eggs. Extraction of eggs with ammoniacal acetonitrile was followed by liquid–liquid defatting, solvent evaporation, and redissolution in a small volume of buffer. The fluoroquinolones were further purified by on-line microdialysis, concentrated on a trace enrichment column, and separated by reversed-phase liquid chromatography with fluorescence detection. Norfloxacin (NOR), ciprofloxacin (CIP), and sarafloxacin (SAR) were extracted from fortified eggs over a range of 2–200 μg/kg, with recoveries of 65.7–78.9%, 65.6–77.1%, and 67.6–110%, respectively. Enrofloxacin (ENRO) was extracted over a range of 1–100 μg/kg, with recoveries of 71.5–86.7%, whereas desethylene ciprofloxacin (DCIP) and danofloxacin (DANO) were extracted over a range of 0.2–20 μg/kg, with recoveries of 68.7–90.7% and 76.0–93.8%, respectively. The limits of quantitation for the 6 fluoroquinolones were as follows: DCIP and DANO, 0.3 μg/kg; ENRO, 1 μg/kg; NOR and CIP, 2 μg/kg; and SAR, 3 μg/kg. Both SAR and ENRO incurred eggs were also successfully analyzed using this method.

Much attention has been paid to the role of developmental information in estimating phylogenetic relationships and, more recently, to the use of phylogenies in understanding the evolution of development. At the moment, however, we lack a sufficiently general theory connecting phylogenetic patterns of character evolution to properties of developmental systems. Here we outline a simple model relating homoplasy to the rate of character change and the number of evolvable states, both of which may reflect developmental constraints. Given a particular rate of character change, the fewer the evolvable states the more homoplasy is expected, and vice versa. The repeated evolution of a limited number of forms of bilateral flower symmetry may reflect constraints imposed by overall flower orientation and underlying mechanisms of differentiation.

Recent phylogenetic analyses have placed the root of flowering plants near Amborella trichopoda, a woody plant restricted to cloud forest habitats in New Caledonia. A distinctive feature of A. trichopoda is its reported lack of xylem vessels. Here we present observations of pit membrane structure and end wall morphology for primary and secondary tracheary cells of A. trichopoda as well as field measurements of stem hydraulic properties of A. trichopoda compared with five cloud forest species from New Caledonia. Observations of stem radial sections revealed that the primary wall material in the protoxylem and metaxylem elements was intact. No large porosities (such as those that have been observed in the pit membranes of Nymphaeales) were observed. However, a few elliptical pits of tracheary cells in the secondary xylem appeared to lack pit membranes. These observations are consistent with our measurements of functional conduit length, which indicate that the longest open conduits are equal to the length of two secondary xylem elements joined end to end. Thus, the xylem of A. trichopoda appears to be functionally vesselless, with the caveat that connections between individual vascular elements may occasionally be open (i.e., lacking in at least one pit membrane). Sapwood area and leaf area–specific hydraulic conductivities of A. trichopoda are similar to those of conifers and angiosperms, with and without xylem vessels, growing in understory cloud forest environments. These findings bear on discussions of the morphology and ecology of the first flowering plants as well as on the possible causes of their diversification.

The fibroblast growth factor receptor (FGFR) family members mediate a number of important cellular processes, and are mutated or overexpressed in several forms of human cancer. Mutation of Lys650-->Glu in the activation loop of the FGFR3 kinase domain causes the lethal human skeletal disorder thanatophoric dysplasia type II (TDII) and is also found in patients with multiple myeloma, bladder and cervical carcinomas. This mutation leads to constitutive activation of FGFR3. To compare the signaling activity of FGFR family members, this activating mutation was generated in FGFR1, FGFR3, and FGFR4. We show that the kinase domains of FGFR1, FGFR3, and FGFR4 containing the activation loop mutation, when targeted to the plasma membrane by a myristylation signal, can transform NIH3T3 cells and induce neurite outgrowth in PC12 cells. Phosphorylation of Shp2, PLC-gamma, and MAPK was also stimulated by all three 'TDII-like' FGFR derivatives. Additionally, activation of Stat1 and Stat3 was observed in cells expressing the activated FGFR derivatives. Finally, we demonstrate that FGFR1, FGFR3, and FGFR4 derivatives can stimulate PI-3 kinase activity. Our comparison of these activated receptor derivatives reveals a significant overlap in the panel of effector proteins used to mediate downstream signals. This also represents the first demonstration that activation of FGFR4, in addition to FGFR1 and FGFR3, can induce cellular transformation. Moreover, our results suggest that Stat activation by FGFRs is important in their ability to act as oncogenes.