THE EGG WHITE PROTEIN EVIDENCE FOR RATITE AFFINITIES

The egg white proteins of the large ratites ( Struthio, Casuarius, Dromaius, Rhea ), the kiwis ( Apteryx ) and several tinamous (Tinamidae) were compared with one another and representatives of several other groups of birds using the technique of isoelectric focusing in acrylamide gel. The tryptic peptides of the ovalbumins of the same groups were compared by thin-layer electrophoresis. The results indicate that the large ratites are more closely related to one another than any one of them is to any other living bird; that the kiwis are not closely related to any of the other groups with which they were compared; and that the tinamous are not closely related to any of the large ratites but may be distantly related to the Galliformes.     

Evolutionary shifts in the melanin‐based color system of birds

Melanin pigments contained in organelles (melanosomes) impart earthy colors to feathers. Such melanin‐based colors are distributed across birds and thought to be the ancestral color‐producing mechanism in birds. However, we have had limited data on melanin‐based color and melanosome diversity in Palaeognathae, which includes the flighted tinamous and large‐bodied, flightless ratites and is the sister taxon to all other extant birds. Here, we use scanning electron microscopy and spectrophotometry to assess melanosome morphology and quantify reflected color for 19 species within this clade. We find that brown colors in ratites are uniquely associated with elongated melanosomes nearly identical in shape to those associated with black colors. Melanosome and color diversity in large‐bodied ratites is limited relative to other birds (including flightless penguins) and smaller bodied basal maniraptoran dinosaur outgroups of Aves, whereas tinamous show a wider range of melanosome forms similar to neognaths. The repeated occurrence of novel melanosome forms in the nonmonophyletic ratites suggests that melanin‐based color tracks changes in body size, physiology, or other life history traits associated with flight loss, but not feather morphology. We further anticipate these findings will be useful for future color reconstructions in extinct species, as variation in melanosome shape may potentially be linked to a more nuanced palette of melanin‐based colors.     

The anatomy of the middle ear of the tinamiformes (Aves: Tinamidae)

The morphology of the middle ear region including the basicranium and quadrate of tinamous is compared among ratites and flying birds belonging to the Procellariiformes, Sphenisciformes, Pelecaniformes, and Ciconiiforms. The middle ears of tinamous and ratites share a number of important characters including absence of a separate foramen for the glossopharyngeal nerve; eustachian tube, carotid artery, and stapedial artery encased in bone; and a metotic process with vascular canals or notches. Outgroup analysis confirms these characters as synapomorphies. These data support the position that the Tinami and Ratiti form a monophyletic assemblage.     

The mating systems of ratites and tinamous: an evolutionary perspective

The breeding behaviour of ratites and tinamous is reviewed. This group includes many of the homeothermic animals which habitually show prominent or exclusive paternal care of eggs and offspring. This unusual parental care pattern is associated with a diverse array of mating systems, ranging from monogamy to mixed polygyny/polyandry. This latter system, typical of the rhcas, is unknown among higher vertebrates outside the taxa considered herein. This diversity of mating systems, together with their great geographical and ecological range, makes ratites and tinamous a group of great potential importance in the investigation of the adaptive significance of social organization, mating systems and parental care patterns. Inadequately described features of their reproductive biology have become incorporated into various considerations of the evolution of reproductive behaviour patterns, and are in danger of assuming the status of fact through repetition. We show that these birds are very little known, in the main, and urge that caution be exercised in the use of what information is available. Directions for new research in ratite breeding biology are suggested, and an interpretation of parental care and pair-bond patterns in these birds is offered.     

Microspectrophotometry of the photoreceptors of palaeognathous birds — the emu and the tinamou

Summary With the aid of a microspectrophotometer the visual pigments and oil globules in the retina of the emu (Dromiceius novae-hollandiae), the brushland tinamou (Nothoprocta c. cinerascens) and the Chilean tinamou (Nothoprocta perdicaria sanborni) were characterized. All three of these palaeognathous birds contain in their rods a typical rhodopsin with _max near 500 nm. Each of these birds has cones containing iodopsin-like visual pigments with _max in the 560–570 nm spectral region. No unequivocal evidence was obtained for the presence of cone pigments other than this iodopsin-like pigment, although one cell thought to be a cone, and containing a visual pigment with _max near 498 nm, was observed in the retina of the brushland tinamou. The oil globule systems of the three palaeognathous species are identical to each other and are much simpler than is typical for neognathous birds in that only two different types of globule are present, one with _T50 at 508 nm and another with _T50 at 568 nm. Comparison of the data with observations made on neognathous species indicates (1) that palaeognathous birds probably have poorer color discrimination capabilities than neognathous birds and (2) that the tinamou is more closely related to the ratites than to the galliform species.This study was supported, in part, by NIH Grant No. EY01839 (A.J. Sillman), NIH Grant No. EY00323 (W.N. McFarland) and NSF Grant No. 78-07657 (E.R. Loew). The authors thank E. Clinite, R. Dunford, C. Murphy, R. Riis and D. Weathers for their valuable assistance. Thanks also go to R.E. Burger for his gift of the emus.     

Diverse stages of sex-chromosome differentiation in tinamid birds: evidence from crossover analysis in <Emphasis Type="Italic">Eudromia elegans</Emphasis> and <Emphasis Type="Italic">Crypturellus tataupa</Emphasis>

All extant birds share the same sex-chromosome system: ZZ males and ZW females with striking differences in the stages of sex-chromosome differentiation between the primitive palaeognathus ratites and the large majority of avian species grouped within neognaths. Evolutionarily close to ratites is the neotropical order Tinamiformes that has been scarcely explored regarding their ZW pair morphology and constitution. Tinamous, when compared to ratites, constitute a large group among Palaeognathae, therefore, exploring the extent of homology between the Z and W chromosomes in this group might reveal key features on the evolution of the avian sex chromosomes. We mapped MLH1 foci that are crossover markers on pachytene bivalents to determine the size and localization of the homologous region shared by the Z and W chromosomes in two tinamous: Eudromia elegans and Crypturellus tataupa. We found that the homologous (pseudoautosomal) region differ significantly in size between these two species. They both have a single recombination event on the long arm of the acrocentric Z and W chromosomes. However, in E. elegans the pseudoautosomal region occupies one-fourth of the W chromosome, while in C. tataupa it is restricted to the tip of the long arm of the W. The W chromosomes in these two species differ in their heterochromatin content: in E. elegans it shows a terminal euchromatic segment and in C. tataupa is completely heterochromatic. These results show that tinamous have ZW pairs with more diversified stages of differentiation compared to ratites. Finally, the idea that the avian proto-sex chromosomes started to diverge from the end of the long arm towards the centromere of an acrocentric pair is discussed.     

The early history of modern birds inferred from DNA sequences of nuclear and mitochondrial ribosomal genes

The traditional view of avian evolution places ratites and tinamous at the base of the phylogenetic tree of modern birds (Neornithes). In contrast, most recent molecular studies suggest that neognathous perching birds (Passeriformes) compose the oldest lineage of modern birds. Here, we report significant molecular support for the traditional view of neognath monophyly based on sequence analyses of nuclear and mitochondrial DNA (4.4 kb) from every modern avian order. Phylogenetic analyses further show that the ducks and gallinaceous birds are each other's closest relatives and together form the basal lineage of neognathous birds. To investigate why other molecular studies sampling fewer orders have reached different conclusions regarding neognath monophyly, we performed jackknife analyses on our mitochondrial data. Those analyses indicated taxon-sampling effects when basal galloanserine birds were included in combination with sparse taxon sampling. Our phylogenetic results suggest that the earliest neornithines were heavy-bodied, ground-dwelling, nonmarine birds. This inference, coupled with a fossil bias toward marine environments, provides a possible explanation for the large gap in the early fossil record of birds.     

Phylogenetic reconstruction of parental-care systems in the ancestors of birds

Due to the controversy surrounding incipient avian parental care, ancestral parental care systems were reconstructed in a phylogeny including major extant amniote lineages. Using two different resolutions for the basal avian branches, transitions between the states no care, female care, biparental care and male care were inferred for the most basal branches of the tree. Uniparental female care was inferred for the lineage to birds and crocodiles. Using a phylogeny where ratites and tinamous branch off early and an ordered character-state assumption, a transition to biparental care was inferred for the ancestor of birds. This ancestor could be any organism along the lineage leading from the crocodile-bird split up to modern birds, not necessarily the original bird. We discuss the support for alternative avian phylogenies and the homology in parental care between crocodiles and birds. We suggest that the phylogenetic pattern should be used as a starting point for a more detailed analysis of parental care systems in birds and their relatives.     

Slow evolution of transferrin and albumin in birds according to micro-complement fixation analysis

Summary Rabbit antisera were prepared to purified ovotransferrin from chicken (order Galliformes) and red-winged blackbird (order Passeriformes) and to purified serum albumin from chicken and rhea (order Rheiformes). Quantitative microcomplement fixation was used to compare these proteins immunologically with those of representatives of all 27 orders of birds. The average interordinal immunological distances were 123 units for transferrin and 53 units for albumin.Extensive intraordinal comparisons of transferrin among 51 species within the order Galliformes and 33 species within the order Passeriformes were also carried out. Values ranging from 0–75 immunological distance units were found within each order.Rabbit antisera to purified alligator albumin were also prepared and shown to react with representatives of all 27 orders of birds, the average immunological distance being 166 units.When the data presented here are considered in relation to the fossil record of birds, it appears that transferrin and albumin have evolved more slowly in birds than in other vertebrates. If prevailing interpretations of the fossil record are correct, transferrin has evolved 2–4 times as fast in mammals and snakes as in birds, while serum albumin has evolved about 3 times as fast in mammals, iguanids, crocodilians, and frogs as in birds. Published immunological and sequence comparisons of lysozyme and cytochromec are also consistent with a slower rate of evolution in birds than in other vertebrates. The implications of a general slowdown in the evolution of bird proteins are discussed.This research was supported in part by grants from the National Science Foundation and the National Institutes of Health to A. C. Wilson and fellowship awards to A. H. Brush and R. A. Nolan (from NIH) and A. C. Wilson (from the Guggenheim Foundation). A preliminary account of part of this work was presented at the International Congress of Systematic and Evolutionary Biology in Boulder, Colorado, on August 9, 1973.The following abbreviations are used in this work: RWBB = red-winged blackbird; OT = ovotransferrin; EW = egg white; TE = tissue extract; ND = not done; MY = million years.     

New nuclear evidence for the oldest divergence among neognath birds: the phylogenetic utility of ZENK (i)

To date, there is little consensus concerning the phylogenetic relationships among neognath orders, which include all extant birds except ratites and tinamous. Different data sets, both molecular and morphologic, have yielded radically different and often unresolved ordinal topologies, especially within the neoaves clade. This lack of resolution and ongoing conflict indicates a need for additional phylogenetic characters to be applied to the question of higher-level avian phylogeny. In this study, sequences of a single-copy nuclear gene, ZENK, were used to reconstruct an ordinal-level phylogeny of neognath birds. Strong support was indicated for the oldest divergence within Neognathae; the chicken- and duck-like birds formed a clade that was sister to all other modern birds. In addition, many families of traditional taxonomic orders clustered together in the ZENK tree, indicating the gene's general phylogenetic reliability. However, within the neoaves clade, there was little support for relationships among orders, which is a result similar to all other recent molecular studies of higher-level avian phylogeny. This similarity among studies suggests the possibility of a rapid radiation of the major neoaves lineages. Despite the ongoing lack of neoaves resolution, ZENK's sequence divergence and base composition patterns indicate its general utility as a new phylogenetic marker for higher-level avian systematics.     

Two methods for using period length to study rhythmic phenomena

Summary We have developed two distinct methods of biological rhythm analysis. The procedures are based on existing techniques for analysis of time series, Enright's periodogram and autocorrelation, and both of the new methods use the parameter, period length (), for defining oscillatory phenomena. We empirically evaluated the two types of analyses using real biological data from circadian rhythm studies in salamanders and sparrows.The first method permits us to make a statistical comparison of period lengths between groups of animals in given treatments. This method is useful for data where the signal-to-noise ratio of the suspected rhythm is very low; and the method is not adequate for making a definitive judgment from single animals. It can best be applied to the question of whether a signal is entraining a rhythm or not and to questions of group differences in period length.With the second method, we determined period length versus time. Using this procedure, we took into consideration the observation that the period length of many biological oscillations changes with time. The method is applicable to records from individual animals, and it can be used to compare treatment effects in individual animals. The technique can also be used to answer the common question of whether periodicityper se exists within a defined range in a time series.We thank Dr, T. J. Crovello, Mr. Kilian, Mr. B. Bailey, Mr. E. Kluth, Mr. G. Wyche, and the Notre Dame Computer Center. Support was provided by postdoctoral fellowship (l F02-HD-52858) from NIH to S. Binkley; grants to K. Adler (NSF GB-30547 and NIH FR-07 033-05); and NSF postdoctoral fellowship (GU-2058) and an Indiana Academy of Science grant to D. Taylor. Sparrow data used in this report were gathered while S. Binkley was a graduate student at the U. of Texas in Austin (NIH traineeship 5T01 GM-00836-08) and with funds from an NIH program project grant (HD-03803-02) to M. Menaker.     

Location of the ureteral openings in the cloacas of tinamous, some ratite birds, and crocodilians: a primitive character

Cloacas of 67 avian species, of both sexes, from various habitats and differing dietary habits, were examined macro- and microscopically to investigate possible variation in the location of the ureteral openings. Differing from most birds studied, in adult male Rhea americana and several tinamous species the ureters were found to open into the coprodeum. In these species the urodeum receives only the vas deferens or oviduct. Similarly, in crocodiles Caiman crocodilus yacare, but not in lizards Tropidurus montanus and snakes Crotalus durissus terrificus, the ureters empty into the coprodeum. This similarity between ancient birds (ratites and tinamous) and crocodiles may indicate a primitive character linking reptiles and birds. This unusual position of the ureteral orifice can represent an adaptation to facilitate urine collection into the coprodeum and large intestine. Another possibility is that this variation in ureter position is a male reproductive strategy to avoid the mixture of urine and semen in the cloaca. There were no evident correlations between the location of the ureteral openings and the birds' habitat, diet, or histology of the coprodeal mucosa. The occurrence of a phallus in eight species of birds was detected, as well as a peculiar vascularization related to the coprodeal epithelium of anseriformes. Together, these data add to the scarce information about the morphophysiology of the avian cloaca, and also contribute to clarify avian phylogenetic linkages.     

Congruent Avian Phylogenies Inferred from Mitochondrial and Nuclear DNA Sequences

Recent molecular studies addressing the phylogenetic relationships of avian orders have had conflicting results. While studies using nuclear DNA sequences tend to support traditional taxonomic views, also supported by morphological data [(paleognaths (galloanseres (all other birds)))], with songbirds forming a clade within Neoaves (all other birds), analyses with complete mtDNA genomes have resulted in topologies that place songbirds as one of the earliest-diverging avian lineages. Considering that over half of the extant bird species are songbirds, these different results have very different implications for our understanding of avian evolution. We analyzed data sets comprising nearly 4 kb of mitochondrial DNA (mtDNA) (complete 12S, ND1, ND2, and cytochrome b) plus 600 bp of the nuclear gene c-mos for 15 birds that were chosen to represent all major avian clades and to minimize potential long-branch attraction problems; we used a partition-specific maximum likelihood approach. Our results show congruence with respect to the ingroup among phylogenies obtained with mtDNA and the nuclear gene c-mos, separately or combined. The data sets support a traditional avian taxonomy, with paleognaths (ratites and tinamous) occupying a basal position and with songbirds more derived and forming a monophyletic group. We also show that, for mtDNA studies, turtles may be a better outgroup for birds than crocodilians because of their slower rate of sequence evolution.     

Early Cretaceous Enantiornithine Birds (Aves,Ornithothoraces) and Establishment of the Ornithuromorpha Morphological Type

New data on the taxonomic and morphological diversity of Early Cretaceous Enantiornithes are reviewed. A new hypothesis concerning the phylogenetic position of Pengornithidae is proposed. These birds traditionally treated as primitive enantiornithines may in fact be more closely related to Ornithuromorpha. This phylogenetic placement implies that the fan-shaped tail and modern-type humeral joint was formed once in the early evolution of birds. Hence, the similarity between Pengornithidae and other enantiornithines may be plesiomorphic. The ecology of Early Cretaceous enantiornithines is discussed. The increased mobility of neck in Holbotia is possibly accounted for by the cranioinertial swallowing mechanism, as in modern ratites. The hypotheses of scansorial adaptations in Parapengornis and Fortunguavis are criticized. In addition, the phylogenetic position of Mystiornis is discussed.     

Studying de-implementation in health: an analysis of funded research grants

Background

Studying de-implementation—defined herein as reducing or stopping the use of a health service or practice provided to patients by healthcare practitioners and systems—has gained traction in recent years. De-implementing ineffective, unproven, harmful, overused, inappropriate, and/or low-value health services and practices is important for mitigating patient harm, improving processes of care, and reducing healthcare costs. A better understanding of the state-of-the-science is needed to guide future objectives and funding initiatives. To this end, we characterized de-implementation research grants funded by the United States (US) National Institutes of Health (NIH) and the Agency for Healthcare Research and Quality (AHRQ).

Methods

We used systematic methods to search, identify, and describe de-implementation research grants funded across all 27 NIH Institutes and Centers (ICs) and AHRQ from fiscal year 2000 through 2017. Eleven key terms and three funding opportunity announcements were used to search for research grants in the NIH Query, View and Report (QVR) system. Two coders identified eligible grants based on inclusion/exclusion criteria. A codebook was developed, pilot tested, and revised before coding the full grant applications of the final sample.

Results

A total of 1277 grants were identified through the QVR system; 542 remained after removing duplicates. After the multistep eligibility assessment and review process, 20 grant applications were coded. Many grants were funded by NIH (n?=?15), with fewer funded by AHRQ, and a majority were funded between fiscal years 2015 and 2016 (n?=?11). Grant proposals focused on de-implementing a range of health services and practices (e.g., medications, therapies, screening tests) across various health areas (e.g., cancer, cardiovascular disease) and delivery settings (e.g., hospitals, nursing homes, schools). Grants proposed to use a variety of study designs and research methods (e.g., experimental, observational, mixed methods) to accomplish study aims.

Conclusions

Based on the systematic portfolio analysis of NIH- and AHRQ-funded research grants over the past 17 years, relatively few have focused on studying the de-implementation of ineffective, unproven, harmful, overused, inappropriate, and/or low-value health services and practices provided to patients by healthcare practitioners and systems. Strategies for raising the profile and growing the field of research on de-implementation are discussed.

     

Complete mitochondrial DNA genome sequences of extinct birds: ratite phylogenetics and the vicariance biogeography hypothesis

The ratites have stimulated much debate as to how such large flightless birds came to be distributed across the southern continents, and whether they are a monophyletic group or are composed of unrelated lineages that independently lost the power of flight. Hypotheses regarding the relationships among taxa differ for morphological and molecular data sets, thus hindering attempts to test whether plate tectonic events can explain ratite biogeography. Here, we present the complete mitochondrial DNA genomes of two extinct moas from New Zealand, along with those of five extant ratites (the lesser rhea, the ostrich, the great spotted kiwi, the emu and the southern cassowary and two tinamous from different genera. The non-stationary base composition in these sequences violates the assumptions of most tree-building methods. When this bias is corrected using neighbour-joining with log-determinant distances and non-homogeneous maximum likelihood, the ratites are found to be monophlyletic, with moas basal, as in morphological trees. The avian sequences also violate a molecular clock, so we applied a non-parametric rate smoothing algorithm, which minimizes ancestor-descendant local rate changes, to date nodes in the tree. Using this method, most of the major ratite lineages fit the vicariance biogeography hypothesis, the exceptions being the ostrich and the kiwi, which require dispersal to explain their present distribution.     

The tritocerebral commissure giant (TCG): A bimodal interneurone in the locust,Schistocerca gregaria

Summary Maturation of the reproductive tract ofLimax maximus was studied in a natural population using a reproductive organ-to-body weight index. Male-phase maturation, denoted by enlargement of the hermaphrodite gland (gonad), took place mainly in June and July. Female-phase maturation, signaled by growth of the albumen gland, occurred primarily in September. When slugs were subjected to artificial photoperiods consisting of long (LD 168) or short (LD 816) days, female-phase maturation was not found to be significantly affected by photoperiod (Fig. 4). In contrast, male-phase maturation appeared to be induced by a short-day to long-day transition (Fig. 7). Implications of this finding for normal seasonal maturation are discussed and a maturation sequence involving both photoperiodic and endocrine control mechanisms is suggested.This work was supported in part by grants from NSF (BNS 76-10783) and NIH (MH 27948) to P.G.S.     

Congruency of phylogenies derived from different proteins

Summary This communication examines the question of phylogenetic congruency- i.e., whether or not the branching order of evolutionary trees is independent of the protein studied. It was found that trees constructed for birds on the basis of immunological comparison of their transferrins, albumins, and ovalbumins agree approximately with a published tree based on the amino acid sequences of their lysozymesc. This congruency is especially noteworthy with respect to the phylogenetic position of the chachalaca, a Mexican bird classified on morphological grounds in the family Cracidae of the order Galliformes. At the protein level, this species differs as much from non-cracid galliform birds as does the duck, which belongs to another order. Despite the organismal similarity between cracid and non-cracid galliform birds, the molecular relationship is remote. If this contrast between organismal and molecular results had been based on comparative studies with only lysozyme, one could have ascribed the contrast to the possibility that chachalaca lysozyme was paralogous, rather than orthologous, to the other bird lysozymesc. Examination of several proteins is thus desirable in cases of possible paralogy.This work was supported in part by grants GB-42028X from NSF and GM-21509 from NIH     

Multivalued stimulus-response relation in isolated elasmobranch utricles

The relation between a maintained spatial orientation and the corresponding fully adapted discharge rate was multivalued in all the afferents tonically sensitive to maintained spatial orientation observed in isolated utricles of Rhinobates productus. The spread of rate values was of the order of changes produced by natural tilts. The occurrence of multivaluedness in isolated receptors indicated that peripheral issues are sufficient. Two factors contributed: firstly, the side from which the orientation had been reached (i.e. hysteresis): higher adapted rates occurred when the preceding orientation was characterized by lower rates and when the corresponding transition caused acceleration; secondly, spontaneous rate variations, some of which resembled markedly, and interacted with, the effects of tilts. It was not possible to identify the basic mechanisms underlying these factors. The multivaluedness in the coding of maintained position, because of its constancy and magnitude, cannot be ignored. It, as well as the sensitivity to fast transients, must be taken into account in utricular models, in evaluations of information transmission, and in psychophysical explorations.Supported by grants (to J.P.S.) from NIH and NSF, and by NIH fellowships (to G.E.W. and R.B.) The tilting equipment was built with funds from the United Cerebral Palsy Foundation