Evolutionary novelties often appear by conferring completely new functions to pre-existing structures or by innovating the mechanism through which a particular function is performed. Sound production plays a central role in the behavior of frogs, which use their calls to delimit territories and attract mates. Therefore, frogs have evolved complex vocal structures capable of producing a wide variety of advertising sounds. It is generally acknowledged that most frogs call by moving an air column from the lungs through the glottis with the remarkable exception of the family Pipidae, whose members share a highly specialized sound production mechanism independent of air movement.
Here, we performed behavioral observations in the poorly known African pipid genus Pseudhymenochirus and document that the sound production in this aquatic frog is almost certainly air-driven. However, morphological comparisons revealed an indisputable pipid nature of Pseudhymenochirus larynx. To place this paradoxical pattern into an evolutionary framework, we reconstructed robust molecular phylogenies of pipids based on complete mitochondrial genomes and nine nuclear protein-coding genes that coincided in placing Pseudhymenochirus nested among other pipids.
We conclude that although Pseudhymenochirus probably has evolved a reversal to the ancestral non-pipid condition of air-driven sound production, the mechanism through which it occurs is an evolutionary innovation based on the derived larynx of pipids. This strengthens the idea that evolutionary solutions to functional problems often emerge based on previous structures, and for this reason, innovations largely depend on possibilities and constraints predefined by the particular history of each lineage.
As noticed by Darwin , every derived feature in an organism must have evolved from a pre-existing feature in its ancestors. Therefore, the current form and function of organism attributes are determined, to a great extent, by phyletic heritage of past events . This equally applies to evolutionary key innovations, which are not designed every time anew, but use available materials, that are themselves a product of millions of years of evolution. This has been firmly established by many studies showing that (i) novel morphological structures may often appear by deploying ancient genetic regulatory circuits [e.g. ] and (ii) existing genes or morphological structures can be recruited to perform completely new functions [e.g. [4, 5]] or to explore new approaches to carry out the same task . Evolution is an integrated and unitary process , and effective reuse of previous molecular or morphological structures through natural selection is subjected to historical constraints . Deciphering how a particular functional mechanism arises in an organism requires framing the question within an evolutionary context through a multidisciplinary approach  involving functional, morphological, and phylogenetic analyses that allow determining which morphological structures were involved, their evolutionary origin (i.e. homology), and the succession of steps that led to a successful end.
Sound production is a key feature in the behavior of different animals such as crickets, frogs, birds or bats . Calls normally serve as advertising signals to delimit territories or attract mates, and in more sophisticated cases can become a proxy of the mood of the individual . Thus, it is not surprising that such a critical function has been the subject of intensive selection through evolutionary history, and a wide variety of sound production mechanisms have evolved in different animals. In frogs, diversification of sound production mechanisms is intimately linked to and/or constrained by the evolution of vocal structures, which is necessarily connected to the evolution of the respiratory system. Despite the considerable diversity of calls and larynx morphologies among extant frogs, the majority of the species call by moving air from the lungs through the glottis . In most frog species, the laryngeal apparatus, which is suspended between the posteromedial processes of the hyoid (= thyrohyals), is a cartilaginous capsule composed of two arytenoid cartilages (each bearing one vocal cord), the cricoid cartilage and associated musculature .
A remarkable exception to the above-described general sound production and larynx morphological patterns occurs in the family Pipidae. The extant members of this family include the South American genus Pipa (Surinam toads) and the four African genera Hymenochirus, Silurana, Xenopus, and Pseudhymenochirus (African clawed frogs). The family Pipidae together with its sister group, the monotypic family Rhinophrynidae (Mexican burrowing toads, genus Rhinophrynus), form the superfamily Pipoidea . The origin of pipids dates back at least to the Mesozoic [e.g. [13–16]] with known fossils from the Cretaceous . Pipids represent a nice example of highly adapted form and function that evolved from an inherited frog bauplan, which is per se highly specialized within amphibians (and tetrapods), and restricted to limited variation . Pipids are the only fully aquatic group of frogs, and their derived morphology and biology are largely a product of adaptations to this lifestyle . One of these remarkable adaptations is the pipid sound production , with the structure and function of their larynx being radically different from those of other frogs [20, 21]. Pipids lack vocal cords, and their larynx is a greatly enlarged and (at least partially) ossified box made up by the cricoid cartilage and the tyrohyals, which do not form part of the larynx in non-pipid frogs. This box encloses the arytenoid cartilages which are modified into two bony rods . The sound production mechanism was described in detail for Xenopus borealis [20, 22], and it appears to be based on implosion of air into a vacuum formed by rapidly moving disk-like enlargements of the arytenoids. The sound is then amplified by the enlarged voice box that serves as an internal vocal sac [20, 22]. Sounds thus are produced without moving an air column, and therefore without externally visible movements of the flanks or throat. Similar motionless calling was also observed in Hymenochirus boettgeri , Pipa pipa , Pipa carvalhoi , Xenopus laevis , and most other pipids [[26, 27], pers. obs.]. However, Pseudhymenochirus was stated to produce sounds by a more conventional sound production mechanism based on moving air , although this behavior has so far not been documented in detail.
Despite their many derived features, in several respects pipids have been more extensively studied than any other group of frogs because Xenopus laevis and Silurana tropicalis have been used as model organisms in physiology, development, and cell and molecular biology [e.g., ]. Knowledge on the closest relatives of model organisms is crucial to interpret and understand the evolutionary origin of studied characters and functions, but remarkably the phylogenetic relationships of pipids have not been comprehensively assessed so far. The rather aberrant morphology of pipids was initially considered to be relatively ancestral among frogs, and many of pipid morphological characters were initially assumed to retain plesiomorphic states. However, now pipids are viewed as highly derived frogs  with many autapomorphies primarily related to their fully aquatic lifestyle . Almost all possible alternative phylogenetic relationships among pipid genera have been recovered based on either morphological [19, 29, 30] or molecular [13, 31, 32] data sets, and the position of Pipoidea with respect to all other frog lineages remains also equally contentious [13–16, 31, 33]. See Additional file 1 for a detailed discussion of previously proposed hypotheses.
Here, we analyze DNA sequences of complete mitochondrial genomes and of nine nuclear genes to produce a robust phylogeny of extant pipoids. We used this phylogenetic framework to gain insights on the evolution of the sound production mechanism in pipids. In this context, we show through behavioral observations that the calling mechanism of Pseudhymenochirus clearly involves the movement of an air column, as it occurs in non-pipid ancestors. Given the unambiguous derived position of Pseudhymenochirus within pipid phylogeny, a reversal to air-driven sound production in this genus is hypothesized. In addition, we provide strong morphological evidence from comparing diverse alizarin-stained frog larynges, which show that larynx structure in Pseudhymenochirus has clear pipid affinities. These observations taken together, allow us to suggest that the use of air in the sound production in Pseudhymenochirus is an evolutionary novelty that evolved by deploying the typical larynx structures of pipids.
We determined for the first time the complete nucleotide sequence of the light (L) strand of the mitochondrial (mt) genome of four pipoid frog species: Hymenochirus boettgeri (HM991331), Pipa carvalhoi (HM991332), Pseudhymenochirus merlini (HM991333), and Rhinophrynus dorsalis (HM991334). The mt genome of Xenopus laevis was the first ever determined for an amphibian  and contained numerous minor sequencing errors probably due to technical constraints at that time. We therefore sequenced anew the full mitochondrial genome of this model species as well (HM991335). Like most metazoans , all five pipoid mt genomes encoded for two rRNAs, 22 tRNAs and 13 protein-coding genes, and conformed to the consensus gene order for vertebrates [35, 36]. All tRNAs could be folded into the typical cloverleaf secondary structure with the known exception of trnS-(AGY). The putative origin of replication of the L-strand (OL) was located between the trnN and trnC genes, and had the potential to fold into a stem-loop secondary structure. Three conserved sequence blocks (CSB-1, CSB-2, CSB-3) were identified in the 3' end of the mitochondrial control region in all pipoid species. Notably, our sequence of Rhinophrynus (a specimen from Tenexpa, Pacific coast of Mexico) differs from a previously determined sequence (GenBank accession number DQ283109; from the Caribbean coast of Texas, US) by a high uncorrected pairwise divergence of 9.9%, suggesting the existence of an unrecognized species in this monotypic genus and family.
Newly generated sequences of partial nuclear genes were deposited in GenBank under accesion numbers HM998927-HM998951, HM998953-HM998985 and HQ260710-HQ260712. Files containing the alignments of both the mitochondrial and nuclear datasets can be accessed in the Dryad Digital Repository under doi:10.5061/dryad.8962.
Maximum likelihood and Bayesian inference methods of phylogenetic reconstruction recovered fully congruent tree topologies for mitochondrial, nuclear, and combined datasets, respectively, with differences only in branch lengths and levels of support (Figure 1). Five major clades were recovered within Anura (Figure 1): Amphicoela (Leiopelma + Ascaphus, which were used to root the tree), Discoglossoidea, Pipoidea, Pelobatoidea and Neobatrachia. Non-neobatrachian frogs were recovered as successively branching lineages, with Discoglossoidea branching off after Amphicoela, followed by Pipoidea and Pelobatoidea. These relationships received high support values in the analysis of mitochondrial genomes and nuclear genes, and maximum support in the combined analysis (Figure 1 and Additional file 1, Figure S1). Alternative phylogenetic placements of the Pipoidea were significantly rejected by the AU test (Table 1 and Additional file 1, Table S2).
Phylogenetic relationships among frogs. ML trees based on concatenated DNA sequences of (a) complete mt genomes and (b) mt genomes plus nine nuclear genes. Numbers at nodes are support values from maximum likelihood bootstrap (1000 replicates; in percent) and Bayesian posterior probabilities, respectively. A congruent topology was obtained by analysis of nuclear genes only (see Additional file 1, Figure S1).
Results of approximately unbiased (AU) tests based on the combined dataset of mitochondrial and nuclear DNA sequences. P-values <0.05 (bold italics) indicate that the data allow rejection of the respective alternative hypothesis.
Within the Pipoidea, all data sets and phylogenetic analyses supported Rhinophrynus as the sister taxon of monophyletic Pipidae, Pipa as sister group to all other extant pipid genera, and sister-group relationships between Xenopus and Silurana, and between Hymenochirus and Pseudhymenochirus. Alternative hypotheses could be significantly rejected except for a basal placement of the Xenopus/Silurana clade (Table 1). In single-gene analyses of nuclear data, Pipa was recovered as sister group to all other extant pipid genera by bdnf, pomc, cxcr-4, slc8a1 and slc8a3, whereas rag1 and rag2 recovered the Xenopus/Silurana clade or the Hymenochirus/Pseudhymenochirus clade in such position, respectively (Additional file 1, Table S1).
In contrast to previous non-documented observations , we provide compelling behavioral data on Pseudhymenochirus merlini showing that this species, while calling, moves a column of atmospheric air from the lungs through the glottis (Additional file 2: Movie). We conclude that this movement of air almost certainly is causal for sound production in this species. Unlike all other extant pipid genera, all of which show a motionless calling, vocalizations in Pseudhymenochirus are clearly associated with intermittent constrictions of the posterior flanks and extension of the throat (Figure 2). The observed sequence of movements further suggests that sounds are produced during expiration, i.e., movement of the air from the lungs (Figure 2). Moreover, males produce release calls, showing also regular contractions of flanks and extension of throat.
Sound production in Pseudhymenochirus merlini. (a) Time series of emission of one note in a male, showing movement of throat and flanks indicative of movement of an air column (also see Aditional file 2: Movie). (b) Spectrogram and oscillogram of a male advertisement call with five notes. (c) Reconstruction under ML of ancestral character states of sound production mechanism (red without, and blue, with movement of air column) using BayesMultistate. (d) Preferred ancestral character state reconstruction of origin (red bar) and reversal (blue bar) of sound production mechanism; white bars represent the less parsimonious hypothesis of three independent origins of the implosion mechanism. (e) Same reconstruction under the alternative pipid phylogeny suggested by morphology.
We complemented behavioral observations with morphological comparative analyses of the larynx structures of alizarin red-alcian blue stained specimens of different pipid genera and a discoglossoid. Our results confirm previous works [29, 37, 38] that larynges of pipids are highly enlarged and ossified, in contrast to those of the rest of frogs. The larynx of Xenopus, is a highly ossified box made up by the thyrohyals, arytenoids, and cricoid cartilages, which are greatly expanded posteriorly. In Hymenochirus and Pseudhymenochirus, the larynges show conspicuous and ossified thyrohyals that enclose the smaller arytenoid rods (Figure 3). Both genera additionally share an elongate shape of lungs that reach the inguinal region and are tightly attached to the body wall (Additional file 1, Figure S4). However, there are two conspicuous differences between these two taxa: (i) Pseudhymenochirus has ossified alary processes of the hyoid plate which form rods very similar to the thyrohyals (= posteromedial processes of the hyoid plate) , and (ii) cartilage and calcified structures around the larynx are more extended and form an overall more compact laryngeal "box" structure in Hymenochirus, with calcified strutures lateral to the thyrohyals and extensive cartilage visible in the glottis area (Figure 3). Because all our cleared-and-stained preparations were made from adult specimens that had been sacrificed immediately previous to the clearing and staining procedure, we can exclude that preservation artefacts have caused these differences. Therefore, the larynx of Pseudhymenochirus seems to be more flexible, and this, somehow may allow the air to move through it.
Structure of larynx in Pseudhymenochirus and Hymenochirus . (a) Cleared and stained preparations of the larynx of Hymenochirus boettgeri and (b) of Pseudhymenochirus merlini in dorsal view, showing a generally lower extension of cartilaginous and calcified structures surrounding the larynx in Pseudhymenochirus. L, lungs; AL, alary processes of hyoid plate; AR, arytenoid cartilages; T, thyrohyals (= posteromedial processes of the hyoid plate). Schematic drawings represent main larynx structures in (c) Hymenochirus and (d) Pseudhymenochirus. Colors denote calcified (red) vs non-calcified cartilaginous (blue) structures. Note the calcified alary process in Pseudhymenochirus. Modified from Ridewood  and Cannatella and Trueb .
Parsimony optimization of ancestral character states of sound production mechanism on the recovered hypothesis of pipid phylogenetic relationships, and on the only not significantly rejected alternative phylogenetic hypothesis (Figure 2) supported homoplasy of the air-driven call in Pseudhymenochirus. Because pipid sound production appears to be linked to the adaptation to aquatic environment  and character states in fossil taxa are unknown, we assume its single origin in the ancestor of Pipidae and a subsequent reversal in Pseudhymenochirus, i.e., two transformations. The alternative hypothesis would require assuming independent origin of the implosion mechanism in Pipa, Xenopus + Silurana, and Hymenochirus, and thus three transformations. BayesMultistate reconstructed the ancestral pipid character state as using the implosion mechanism, with a ML probability >0.999 (Figure 2).
Our results provided strong support for (i) the placement of monophyletic pipoids as the third most basal clade among extant anurans and (ii) the monophyly of Pipidae. These results are concordant among all data and methods of phylogenetic inference, and in agreement with previous molecular [13, 14] and morphological [10, 39, 40] studies. Within Pipidae, our results supported the monophyly of dactylethrines (Xenopus and Silurana) as in other molecular studies [14, 32] and recent morphological studies , and significantly rejected the previously proposed paraphyletic basal arrangement of Xenopus and Silurana [19, 29]. The recovered basal position of Pipa, and the sister group relationship of dactylethrines and hymenochirines (Hymenochirus and Pseudhymenochirus) agrees with previously published molecular studies [13, 14] but contests recent morphological analyses [17, 19, 29, 30] that supported a Pipinae clade (including hymenochirines and Pipa) [17, 19, 29] with up to six osteological characters [after excluding fossil taxa; ]. The alternative hypothesis of a Pipinae clade could not be rejected by our molecular data (Table 1 and Additional file 1, Table S2). If our preferred hypothesis of pipid relationships is correct, homoplasy of the above mentioned six characters needs to be assumed. Polarization of these characters is complicated by the fact that all extant pipids are aquatic whereas their unambiguous extant sister group, Rhinophrynus, is a terrestrial species with specialized burrowing habits. Future morphological studies should assess additional external characters, tadpole morphology, and soft anatomy in the search for possible synapomorphies of the four African genera, such as the keratinization of the first three toes, which is more strongly expressed in the African taxa .
Despite the apparent contradiction to morphological data, the congruence between mitochondrial data and the various nuclear genes (Figure 1 and Additional file 1, Table S2), and the strong support for Pipa as sister to all other extant pipids make our conclusions considerably robust. From a biogeographic point of view our hypothesis suggests that the basal split among extant pipids might have separated an African lineage (Hymenochirus, Pseudhymenochirus, Silurana, and Xenopus) from a South American lineage (Pipa) and is consistent with the American distribution of Rhinophrynidae as sister group of the Pipidae [41, 42]. This indicates the need of re-evaluating also the phylogeny of fossil taxa, given the apparent biogeographic anomaly that the South American Pipa based on morphological data are nested within a clade of purely African fossil taxa, and the African Silurana/Xenopus within a clade of exclusively South American fossil taxa .
Our behavioral observations suggest that sound production in Pseudhymenochirus is produced by air movement from the lungs to the throat. This is unique among pipids, which generally produce motionless clicking sounds by implosions related to the derived box-like structure of the larynx , and in fact more similar to the typical mechanism found in non-pipid frogs. The sound production mechanism of pipids has been thoroughly studied in Xenopus borealis  and given the resemblance of calls and motionless calling behavior in other members of the family [21, 23–27], it is assumed to be the general system in pipids. In X. borealis, the characteristic clicking sound was proved to be produced by the simultaneous action of bipennate muscles that separate the discs of ossified arytenoid rods . Yager suggested that the sound is produced by the implosion of air when the two arytenoid discs separate, given that no clicking sound was emitted when this space was filled with liquid . Similarly, the implosion mechanism is not air-driven, because call spectra remained unchanged after frogs were foreced to breathe helium .
With regards to morphological analyses, our results are fully congruent with previous detailed anatomical descriptions in which the larynges of pipids are enlarged boxes formed by more or less ossified cartilages [20, 21, 37, 38]. Despite the apparent diversity in larynx morphology both among pipids and among frogs, the embryological origin of involved cartilages have been traced back to the larval hyobranchial apparatus , leaving little doubt of their homology within amphibians [37, 43]. The larynx of Hymenochirus is an enlarged box with ossified cartilages, more similar to that of Xenopus and Silurana than to other non-pipid anurans (laryngeal cartilages are not ossified, e.g. Bombina in Additional file 1, Figure S5), thus reinforcing a similar sound production system to that of X. borealis. In Pipa, although the larynx structure slightly differs from that of Xenopus [20, 22] a similar sound production has also been suggested . The larynx of Pseudhymenochirus is particularly similar to that of Hymenochirus and shows the typical ossified cartilages of other pipids [37, 38]. Therefore, we could undoubtedly assert that the larynx in Pseudhymenochirus evolved from a typical pipid condition, but the overall structure seems to be more flexible, and this could somehow permit a movement of air that is used to vocalize, as suggested by our behavioral observations. However, whether vocal cords are present in Pseudhymenochirus (which are absent in pipids) or whether other strucutres are responsible for sound production requieres specific examination. Other hypotheses may also be plausible, and further detailed functional studies [as those performed by Yager; ] are needed in order to determine the exact mechanism through which sound is produced, as well as the precise function of involved structures. Overall, our molecular and morphological data leave neither doubt of the nested phylogenetic position of Pseudhymenochirus within Pipidae, nor of the clear pipid nature of its larynx. While the source used for sound production unexpectedly appear to reverse to the ancestral non-pipid condition (movement of the air column), associated anatomy evolved from a typical pipid-like larynx that likely imposed constraints to natural selection. Altogether, we suggest that the air-driven sound production in Pseudhymenochirus most probably represents a novel evolutionary combination and it is a remarkable example of complex anatomical modifications related to a functional shift of enormous influence in frog behavior and life history.
The selective forces for these changes are unknown, but the movements of the body flanks during the call in Pseudhymenochirus obviously produce water waves that might provide information about the size of the calling male to females, detected by their lateral line system. Water surface waves can play an important role in the advertisement behavior of several basal anurans [44, 45]. Compared to Hymenochirus, sexually active Pseudhymenochirus males have morphologically less distinct postaxillary glands (Figure 2a), which are used in chemical communication during the breading season in Hymenochirus . Therefore, flank movements in Pseudhymenochirus could serve as additional visual and mechanical signals, which might reinforce the acoustic signals to attract females and impress conspecific males.
Our study exemplifies that understanding the evolutionary process underlying an innovation, here the air-driven call in Pseudhymenochirus, can only be achieved through an integrative comparative approach. In this particular case, behavioral observations prompted for detailed anatomical analyses, and comparative data were placed within a robust phylogenetic framework based on molecular data. Further insights on the nature of this evolutionary innovation could be gained through ontogenetic studies that disentangle how morphological constraints imposed by the rather stiff larynx box of pipids are overcome to allow the reversal to the ancestral air-driven vocalization in Pseudhymenochirus. The result of this study provides yet another example of how natural selection generates complex morphologies and functions by tinkering with previously available structures , and further reinforces the important roles of historical contingency and constraints in canalizing potential solutions to a given evolutionary problem .
We assembled a dataset of all mitochondrial genomes of frogs available from GenBank, expanded it with four (newly determined) mt genomes of additional pipoid taxa, and replaced the available sequence of Xenopus laevis  by a newly determined one from a specimen with reliable locality data. DNA was extracted using a standard phenol-chloroform protocol from voucher specimen tissue. Several overlapping fragments covering the entire mt genome were amplified by PCR using previously reported primers and cycling conditions . Specific primers were also designed to amplify fragments in some species in which general primers did not work (available from authors upon request). Those fragments that contained the control region were cloned into pGEM-T vectors (Promega, Madison, WI, USA) due to observed heteroplasmy. PCR fragments and recombinant clones were cycle-sequenced with the ABI Prism BigDye Terminator Cycle Sequencing Ready Reaction Kit (V3.0) using PCR and M13 universal primers, respectively, as well as walking primers if needed. Cycle sequencing products were run on ABI Prism 3700 and 3130 × l DNA Analyzers (Applied Biosystems). The obtained mt sequences were annotated based on sequence similarity to reported frog mt genomes. The vertebrate mt genetic code was used to translate ORFs of protein-coding genes. The different tRNAs were identified based on their putative clover-leaf secondary structure, as implemented in the program DOGMA . Sequences were aligned by taking secondary structure of tRNAs and amino acid translations of protein-coding genes into account. Highly variable portions of the sequences, as well as third positions of mt protein-coding genes were excluded from phylogenetic analyses (see Additional file 1 for extended methods).
A nuclear DNA dataset was generated using partial sequences of nine protein-coding genes (see Additional file 1, Table S3): rag1, rag2, bdnf, pomc, exon 2 of cxcr4, exon 2 of slc8a1, slc8a3, exon 1 of rho and H3a. We assembled a complete combined matrix complementing previously available sequences from GenBank with newly determined sequences, representing all major lineages of frogs for which mt genome data exist. In a few cases, chimerical sequences were constructed by merging sequences from different species of the same genera, for which strong evidence exist of being monophyletic. Primers used were as reported in the literature: rag1 ; rag2 [51, 52]; slc8a1 ; bdnf and pomc ; rho ; and H3a . In all cases, PCR cycling conditions were experimentally adjusted from those reported in the original publications.
Single-gene alignments were used to produce three different datasets, containing: (a) all mt genes (final length of 11,131 bp); (b) all nuclear genes (final length of 7,107 bp); and (c) a combination of mt and nuclear genes (final length of 18,238 bp). Additionally, we also constructed an alternative mt dataset using amino acid characters for protein-coding genes, as well as analyzed single-gene datasets for the nuclear genes to understand the congruence among these markers.
We used Leiopelma and Ascaphus as outgroup taxa because molecular and morphological data are congruent in indicating that these are the most basal extant frogs [14, 55]. All datasets were subjected to maximum likelihood [ML; ] and Bayesian inference [BI; ] analyses using RAxML version 7.0.4  and MrBayes version 3.1.2 [59, 60], respectively. RAxML used the rapid hill-climbing algorithm  computing 100 distinct ML trees starting from 100 distinct randomized maximum-parsimony starting trees. BI was performed running four simultaneous Markov chains for 10 million generations, sampling every 1000 generations. An additional BI run was performed for each of the analyses, to confirm the adequate mixing of the Markov chains. Convergence was checked a posteriori by plots of lnL scores and low standard deviation of split frequencies, as well as using the convergence diagnostics implemented in the online tool AWTY . The first 2.5 million generations were discarded as burn-in to prevent sampling before the Markov chains reached stationarity. Partitioned analyses were performed for ML and BI, with 16 partitions for the mitochondrial and 9 for the nuclear datasets. For each partition, the best fit-model of nucleotide substitution was chosen using the Akaike information criterion [AIC; ] as implemented in Modeltest version 3.7 , MrModeltest version 2.3 (by J. A. A. Nylander; http://www.abc.se/~nylander/), and ProtTest . Support for internal branches was evaluated performing 1000 replicates of non-parametric bootstrapping  (ML) and by posterior probabilities (BI).
Alternative tree topologies (see results) were evaluated based on the combined mt and nuclear dataset using the non-parametric approximately unbiased (AU) test  as implemented in Consel version 0.1 k  with site-wise log-likelihoods calculated by RAxML with independent GTR + Γ + I models assigned to each of the different partitions, and one million multiscale bootstrap replicates. We used BayesMultistate  from the BayesTraits package (by M. Pagel and A. Meade; http://www.evolution.rdg.ac.uk/BayesTraits.html) to infer the ancestral state of the mechanism for sound production in the family Pipidae using ML.
In order to further understand the morphological basis of sound production in pipids, we performed a comparative anatomical study of the larynx structures of several pipids including Xenopus laevis, Hymenochirus boettgeri and Pseudhymenochirus merlini, as well as Bombina bombina as representative of non-pipid ancestors. Following standard international procedures, specimens were sacrificed using an overdosis of MS222, fixed in formalin, and differentially stained for bone and cartilage with alizarin red S and alcian blue, respectively, following a standard procedure .
Observations were made of captive specimens in ca. 100 × 30 × 20 cm aquaria. Specimens of different pipid species were obtained from the pet trade and kept at different times between 1985-2011. All observations refer to specimens in breeding conditions, without external (hormone) stimulus. No experiments with living animals were performed. Video sequences of calling specimens of Pseudhymenochirus were recorded in 2010 with a Sony DCR-SR30 camera. Spectral and temporal variables of the recorded sounds were analyzed using Cooledit 96 software (Syntrillium). Sonagrams were constructed using the package seewave  in the R environment .
We are grateful to D. Buckley and M. García-París for insightful discussion on the manuscript and we appreciate the contribution of A. Crottini, S. Esser, D. M. Green, K. Kunz, I. Martínez-Solano and G. Parra-Olea for tissue samples and specimens, and of R.D. Randrianiaina with microscope photography. I.I. was supported by a JAE PhD fellowship of the Consejo Superior de Investigaciones Científicas (CSIC) of Spain. D.S.M. was supported by the EU Marie Curie Mobility and Training Programme (FP7, proposal PIEF-GA-2009-237658). This work received financial support from MICINN to R. Z. (CGL2004-00401).
II carried out molecular lab work. MV provided anatomical data. FG performed behavioral observations. II and DSM analyzed data. II, MV, DSM, FG and RZ wrote the paper. All authors read and approved the final manuscript.


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A New Era for China Investors Aidan Shevlin, Head of Asia Pacific Liquidity Fund Management, J.P. Morgan Asset Management Chinese financial regulators have recently published new rules for China's CNY 100trn asset management industry. Aidan Shevlin, J.P. Morgan Asset Management, outlines the implications of the new rules for the industry and for corporate treasurers - and suggests practical steps for rethinking liquidity management in China.
The final version of the new rules for China's CNY 100trn asset management industry has now been published by the Chinese financial regulators. In this Executive Interview, Aidan Shevlin, Head of Asia Pacific Liquidity Fund Management, J.P. Morgan Asset Management, explains the implications of the new rules for the industry, and for corporate treasurers, whilst outlining practical steps for rethinking liquidity management practices in the country.
What are the new asset management industry rules? When do they come into force in China, and what will their broad impact be?
Officially known as the 'Guiding Opinions on Regulating the Asset Management Business of Financial Institutions', the new regulations represent a comprehensive restructuring of China's asset management industry. Introduced by the new Financial Stability and Development Committee (FSDC), the rules are designed to reduce financial risks, simplify products and increase investor protection.
The new rules came into force on 27 April 2018. As of now, all newly created Asset Management Products (AMPs) must comply with the rules; while existing AMPs have until the end of 2020 to be fully compliant.
At a glance, what are the new AMP rules looking to achieve?
In brief, the new rules are designed to: limit shadow banking activities; ban expected return and principal guarantees; harmonise regulatory standards; and reduce regulatory arbitrage.
What constitutes an 'Asset Management Product' according to the rules?
Historically, shadow banking was the broad term for all non-time deposit products issued and managed by commercial banks and financial institutions, including wealth management products (WMP), trust products (TP), asset management plans and so on. The new regulations specifically define Asset Management Products (AMPs) as an umbrella term for all these instruments (with a few exceptions such as private funds and pension products).
Under the new rules, all public AMPs must be managed on a mark-to-market net asset value (NAV) basis (there are some exceptions for amortised NAVs) without offering expected returns or implicit guarantees. AMPs will have strict rules on leverage, layering and risk reserves; in addition they must be managed by a segregated asset management business and have a separate custodian.
How does the regulator define standard assets and non-standard assets?
The new rules stipulate that publicly-offered AMPs, which make up the majority of existing products, can only invest in standard assets (SAs). Typical standard assets are tradable fixed income instruments or equity shares. Non-standard assets (NSAs) are essentially all other assets.
According to the new rules, qualified investors can still buy privately offered AMPs that invest in NSAs, but there are strict rules regarding the type of investors, pricing of the NSAs and type of investments that private AMPs can put money into. As such, it is worth seeking specialist advice on this topic where appropriate.
Why were new regulations needed for the shadow banking sector and what were the Chinese authorities looking to achieve?
A confluence of factors has led to the introduction of these new rules – but clamping down on China's shadow banking industry has been one of the major drivers. Shadow banking in China is a by-product of past financial regulations and interest rate liberalisation, as borrowers and lenders sought to circumvent rules to achieve access to capital at more market-driven interest rates.
It is not hard to see why shadow banking became so popular: investors benefited from very attractive and stable yields with principal guarantees, while financial intermediaries like commercial banks were able to take assets off-balance sheet and generate profitable fee income; and finally these products offered financing to issuers who were perhaps unable to access other sources of funding. Between March 2015 and September 2017 the amount of shadow banking products outstanding doubled from CNY54.2trn to CNY107.6trn . At their peak in Q4-2015 wealth management products – the largest individual asset class, representing a quarter of assets - grew by 56%y/y .
But in some ways, these benefits were too good to be true. There was a distinct lack of transparency around the underlying assets in many shadow banking products, and in recent years some of these opaque assets turned out to be very poor quality. In addition, the use of leverage and layering created a complex web that touched almost every part of China's financial industry. While certain banks performed bail-outs where necessary, this only fuelled further investor interest, since they assumed that their WMPs were 'safe' – triggering concerns about moral hazard.
By the end of 2017, the shadow banking sector had ballooned to US$12.3trn, representing approximately 93% of China's GDP and approximately 63% of all bank loans . The Chinese authorities realised this behemoth need to be contained while also eliminating misconceptions about principal guarantees and establishing stronger links between risk and return. To this end, the new rules are a big step in the right direction. In fact, the AMP rules are one of the most significant changes ever in the history of China's shadow banking industry.


FireBuilder — Chemong Chimney Ltd.
Please enjoy using Travis Industries' FireBuilder application, featuring their Lopi and Fireplace Xtrordinair products. You can use images of your own home to customize and preview different designs, as well as view technical specifications for different models.
Each product within is available for installation from Chemong Chimney professionals.


Now that the dog days of summer are upon us, there is very little appeal in turning on the stove or the oven. On a sweltering day or evening there is nothing better than a frosty cold and delicious bowl of gazpacho.
Gazpacho originated in Andalusia in Southern Spain. Gazpacho appears to have evolved over the millennia. The earliest form of gazpacho likely arrived in Southern Spain with either the Romans or the Moors and was a peasant soup made from garlic, olive oil, stale bread, salt and vinegar. The tomato-rich gazpacho that we know today did not evolve until the 1800's, centuries after Christopher Columbus first introduced tomatoes from the New World to Europe.
There are various types of traditional gazpacho made throughout Southern Spain. Most include garlic, bread, olive oil, Spanish Sherry vinegar, tomatoes, peppers, onions and cucumber, although there are several variations that do not include tomatoes. But many other variations abound including green gazpachos, white gazpacho, gazpachos made with almonds, mushrooms, asparagus and green grapes.
Traditionally, gazpacho was made by hand with a mortar and pestle. Garlic was pounded with soaked bread, olive oil and salt to make a paste. Tomatoes, vegetables and vinegar were added and pounded until a soup-like consistency was obtained.
Fortunately, today we have food processors that allow us to whip out a batch of gazpacho in 15 - 20 minutes. It really could not be simpler. Start with some garlic and perhaps a small piece of shallot. Whirl until finely minced, then add your fruits and vegetables, some seasoning, perhaps some fresh herbs and a little Spanish sherry vinegar, tomato juice and olive oil. Presto, you have a refreshing meal.
While bread is a traditional gazpacho ingredient, I have developed most of my gazpacho recipes without it to accommodate people on gluten-free or low-carb diets. I have one gazpacho recipe where I have added quinoa to give it a bit of heft, without the gluten. Almonds can be finely ground as an alternative to bread.
The quality of your ingredients matters tremendously with gazpacho. So use ripe produce at its peak. If tomatoes are not great at the moment you are having a gazpacho jones, consider roasting them for extra flavor. Likewise use only great extra-virgin olive oil (avoid lite olive oil like the plague) and Spanish sherry vinegar in your gazpacho. An off tasting oil or vinegar will ruin an otherwise great gazpacho.
Note: I am an Amazon Affiliate and I make a small commission when you purchase items through my links.
3. Pureed or Chunky Gazpacho?
There are two styles of gazpacho: 1) smooth and 2) chunky. In general, I like a little chunk to my gazpacho, but depending on the ingredients, in some cases, like my Sugar Kiss Melon Gazpacho, I will make a smooth gazpacho.
What I find works best is to roughly chop the vegetables in a food processor. Set 1/2 to 3/4 of the chopped vegetables aside and then finely puree the remaining 1/4 to 1/2 of the ingredients. This gives gazpacho a little texture without being too salsa like.
The wonderful thing about gazpacho is it gets better after it has a chance to rest in the fridge. I like to make a big batch and then eat it for lunch all week long.
To really bring the wow factor to your gazpacho, get creative with the garnish. Marcona almonds and a little feta is delicious. Crispy croutons or toasted small toasts baked with a bit of goat cheese add crunch and tang. Diced vegetables such as avocado, peppers and cucumber work well. Chopped boiled egg or some Spanish Iberico jambon are very nice.
My favorite trick is to pack gazpacho in 8 ounce mason jars. I fill the jars and refrigerate. The jars make for individual servings that are great for a packed lunch or a picnic. I think these Italian mason jars are particularly nice. I use them for individual servings of gazpacho, cold soups, shrimp cocktails and desserts. I also use them to make a week's worth of homemade salad dressing.
This Watermelon Gazpacho is fresh and feels like summer in a bowl. I add watermelon and basil leaves to the more traditional gazpacho ingredients and garnish with salted Marcona almonds and a sprinkling of feta cheese.
This summer-ready gazpacho is what you need to make when peach season is at its peak. The goat cheese croutons make for a great crunchy, creamy and tangy garnish.
I discovered this this divine avocado gazpacho at The Vintage Restaurant in Sun Valley, Idaho. We were served this on a hot summer's night and immediately fell in love with it. There was something about the flavor that we could not put our finger on. Fortunately our waitress was willing to clue us in, and Jeff Key's cookbook was available for purchase so I was able to procure his recipe. You can find the recipe and the secret to this tasty soup here.
I developed this recipe a few years ago when I was experimenting with a gluten-free diet. I had a favorite gazpacho recipe by Elizabeth Shepherd, published in Epicurious in May 2001, called Mom's Gazpacho - Best Ever, that called for bread crumbs and chopped boiled eggs. The boiled eggs are mashed with garlic and salt to create a foundation for the soup. The bread crumbs give the gazpacho a bit of heft. I modified Elizabeth's recipe by using quinoa as a substitute for the bread and the result was delicious.
If gazpacho and ceviche had a baby, this recipe would be their progeny. It marries the best of gazpacho and ceviche and is a summertime party favorite at our house. Ingredients include shrimp, crab, tomatoes, cucumber, bell peppers, lime juice, Spanish Sherry vinegar, cilantro and cumin. I use canned crab from Costco to keep the cost down. I also make a big batch and eat it all week long. The flavor get better and better.
This recipe was inspired by a gazpacho I had at Mountain Standard Restaurant in Vail, Colorado. Roasting the peppers and tomatoes creates a smokey goodness and makes for a unique gazpacho.
This is an amazing gazpacho that is perfect for bridging the Summer and Fall seasons. It uses sweet Sugar Kiss melon, toasted pepitas, feta cheese and a sprinkling of pomegranates that begin to come into season in the late summer. I throw in a bit of avocado to create a creamy texture. For this gazpacho I process it in a food processor or a blender to make it silky smooth. The end result is smooth, sweet, crunchy, tangy and totally refreshing.
Lest you think gazpacho is just a tasty summer soup, it is also extraordinarily healthy. Gazpacho fits in well with the Mediterranean diet that focuses on plenty of fresh fruits and vegetables, and lots of olive oil. There are lots of health benefits to adhering to the Mediterranean diet, but a recent study shows sticking with a Mediterranean diet is very good for your brain. University of California San Francisco just presented a study at the Alzheimer's Association International conference in London that shows adhering to a Mediterranean diet results in better cognitive function and 30 - 35% lower risk of age-related cognitive impairment. So dust off your food processor, throw in some fresh produce and a generous pour of extra virgin olive oil. Gazpacho is a delicious way to eat your way to long term brain health.


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BNL Gruppo BNP Paribas
We Love Cinema
Mobile , Web
VIEW SITE:
http://welovecinema.it
We Love Cinema is the BNL project dedicated to those who do, follow, and love cinema, which aims to create the first community of film enthusiasts and insiders.
The evolution of the interpersonal dynamics triggered by social media and mobile technology makes the crucial role of the vertical community increasingly clear, characterized by groups of active users in the production of content and the sharing of specific interests and passions.
From this point of view the aim of BNL was to completely innovate the viewing experience of cinema culture for all fans and insiders through the integrated We Love Cinema platform (website, apps, and social media). With an integrated digital awareness project, we preside over the communities of film fans, and we encourage their direct involvement and the production of content related to the world of cinema (corporate business activities that the bank has overseen since 1935, with funding from the cinema industry).
The telling of the history of cinema through a website and a mobile application (and relaunch through social media) was conceived as a process that aims for the direct involvement of the community of cinema lovers, continuing a tradition that for 80 years has tied BNL to what the critic Ricciotto Canudo defined as the "seventh art."
The initial phase of the project focused on an analysis of users, profiling them by type of interest, main services used and the contributions that they could offer in terms of content and participation. Eurisko data shows that:
- Almost 50% of cinema goers are aged between 25 and 44;
- Film fans typically go to the cinema 6 times a year;
- There is a core target (about 2 million people) that goes to the movies on average 20 times a year, and is worth 40% of the entire market in terms of tickets.
To this type of analysis was added a study by the app store, analyzing in particular the ecosystem of mobile applications dedicated to cinema, from which it emerged that:
- the search term "cinema" creates 2111 results for iPhone apps;
- traffic for the search term " cinema " is medium-high;
- most applications have not conducted a correct ASO and many apps are not even updated frequently.
The acquisition of this data and the analysis of the experiences gained from film fans have allowed us to innovate the viewing experience of cinema culture, providing them with a personalized service in which to fully enjoy their passion.
The previous website was characterized by a content structure that favored consultation, with a low possibility of specific interaction by users. We aimed for a complete overhaul of the site, turning it into a touchpoint that can directly connect the BNL brand with cinema fans.
The CMS has been updated to allow the publication of content by exploiting the full functionality of WordPress, and has been integrated with an API system developed to ensure enjoyment even on Android and iOS mobile applications. The apps have been designed with a focus on three main drivers:
1. Design and implementation of content and information from the world of cinema, in order to ensure maximum usability by both web and mobile platforms;
2. Design of the User Interface and User Experience, to ensure an engaging and rewarding user experience;
3. Constant analysis of the performance of the website and the iOS and Android apps, to ensure scalability and maximum user involvement.
In the mobile application a push notification system was also implemented in order to activate the direct involvement of users on specific occasions.
The We Love Cinema project has today developed into a new fully responsive website and a mobile application that allows all users to be protagonists in an innovative and engaging experience, able to enhance their passion for cinema.
The mobile application and website allow the user, at any time and from any place, to acquire all the latest information and news from the world of film: from reviews written by respected names to insights about film jobs, and including BNL's commitment to the funding of more than 5,000 films, many of which are part of cinema history.
In addition, the "Cinema Locator" feature allows you to find cinemas near you, view the weekly listings, contact them and set your navigator in order to get there. While the Media Gallery and the area dedicated to international Festivals allow you to experience the excitement of the world's most famous red carpets.
The correct settings for App Store Optimization, which have become essential in the launch strategy and continuous updating of a new application, immediately brought about concrete results.
Just one week from the application launch it was selected by Apple as one of the "Best New Apps" and was highlighted in the App Store, visible from both desktop and mobile.
With this project we have consolidated our expertise in developing mobile apps for vertical communities, fundamental today for those who want to activate aggregation dynamics of active users in content production and the sharing of specific interests and passions.
START YOUR DIGITAL TRANSFORMATION
Send us a message and we will get back to you soon!
Registered office: Viale dell'Esperanto 71 – 00144 Roma
VAT registration number 11289201003 - Cap.Soc. 10.000 €
Email: info@iquii.com
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From the bestselling author of An Angel Healed Me and An Angel Called My Name comes a groundbreaking book featuring true stories of afterlife encounters, offering powerful clues for discovering our soul's purpose and unlocking our divine potential.
With advances in modern medicine enabling more and more people to be brought back from the brink of death, it is now possible to envision what lies beyond. It is also possible to discover more about our spiritual destiny through these near death experiences. Each afterlife experience offers powerful clues into our soul's purpose, and in this fascinating new book, Theresa Cheung unveils these life-changing truths, or insights, as the Ten Secrets of Heaven. It is only through our understanding of their true meaning on earth that we can discover heaven and eternal life.
After explaining each of the ten secrets, Theresa reveals how heaven can disguise itself in aspects of daily life, such as health, career, money, love and relationships, and life goals, and also during times of crisis, such as when we are depressed, grieving, lacking direction, or experiencing heartbreak. Truly understanding how the Ten Secrets of Heaven can be harnessed for our wellbeing will bring enlightenment, peace, and joy. It will also leave no doubt that nothing in this life is as it seems—and death is not the end.
Theresa Cheung was born into a family of spiritualists and has been involved in the research of psychic phenomena for over twenty-five years. She has a master's degree from King's College, Cambridge, and has written a variety of books about the paranormal including the Sunday Times bestsellers An Angel Called My Name and An Angel Healed Me. She lives with her husband and two children in Oxford.


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Q: How the SharedPreferences works and is it safe I'm going to use the SharedPreferences Keys to store my app informations so when I open the app again after onDestroy the information will still the same.
I'm just wondering is it safe to use SharePreferences ? I mean is there a way to hack or get the KEYS from the SharePreferences ?
and does all Android Platforms have the SharePreferences  ?
thanks .

A: sharedPreferences arent safe..
sharedPreferences should just store config/setting-data not encrypted..
if u want to store critical data - you have to write it encrypted in a dataBase/sharedPrefs
btw .. http://android-developers.blogspot.de/2013/02/using-cryptography-to-store-credentials.html

A: You shouldn't store any unencrypted valuable information(passwords, private user information etc.) in SharedPreferences. SharedPreferences are just plain XML files in app directory on internal storage.
If you need to store smth private - you definitely need to encrypt it first.

redpajama_set_name: RedPajamaStackExchange


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of my clients into reality.
Imagine living in an area virtually unknown by the mass population yet close enough to commute to any of three large North Carolina cities.
Picture yourself surrounded by nature and breath taking views or living on the shore of sparkling High Rock Lake, second largest lake in NC.
over an acre of woodland and increase according to location, size and view.


We are open for everyday for people to attend services; for private prayer and meditation; for viewing the wonderful architecture and stained glass; to learn about the history of the building and the Minster site; to enjoy the Minster grounds.
Doncaster Minster is within easy reach of the railway station, transport interchange and the town centre. Our near neighbours are Doncaster College, The Frenchgate Centre and Doncaster Market.
We welcome thousands of visitors each year, and we always have some of our friendly stewards on hand to help, support and guide you around the Minster. The main entrance is through the South Porch, once inside you will find a range of information leaflets in several languages to provide you with an introduction to the Minster.
Please note that there may be restricted access during services and events.
Shop: The Minster has a small shop area which sells cards, postcards, souvenirs, guides, CDs and mementos. Take the opportunity to browse what is on offer during your visit.
Photographs: The Minster is free to view but, if you wish to take photographs as a reminder of your visit we ask you to purchase a photo permit (£3). This enables you to take photographs of the Minster (not during Services) for your own use. Please contact the Minster Office if you wish to take photographs for publication or for commercial use.


One each by Bosnian, Czech, Egyptian, Finnish, Irish, and Swedish writers. Plus one book by an author of uncertain nationality (Olaudah Equiano — was he born in Africa or South Carolina?) and three books by multiple authors from various nationalities. There was not as much diversity here as usual, alas.
I'm reading a lot more contemporary fiction lately, which I don't like in theory, although I'm enjoying it in practice.
As for cycling, my total mileage is way down this year, for obvious reasons. I rode 3,677 miles, down from 5,213 the previous year. But I rode over 3,000 of those miles in the first half of the year, mostly before I knew I was pregnant. If I'd kept up that pace, I would have been close to my old yearly mileage record of 6,597. That's a number I won't see again for a while. I did some races last year, maybe 6 or so, but that's all over for a while. Next year, I'll be grateful for every mile I get to ride, and I won't even think about racing.
First, there was Kenzaburo Oe's novel A Personal Matter, which is a strange choice of book to read during pregnancy, since it's about a man who discovers that his son was born mentally handicapped. He spends the rest of the novel reacting badly to this news. But I wasn't bothered by the subject matter, and I liked the novel a lot. There's an unsparing directness to it, a sense of strangeness and a willingness to dig deep into the main character's disturbing, although in moments unexpectedly sympathetic, mind that I admired.
Then I read Tim Parks's illness memoir Teach Us to Sit Still, which I also liked very much. He tells the story of mysterious pelvic pain that he suffered from for many years before feeling desperate enough to seek solutions in unexpected places. He turns to various forms of meditation and finds that this helps him recover and transforms him in deeper ways as well. The book is a really interesting exploration of the limits of western medicine and the surprising (to him and to many other people I'm sure) connections between the mind and the body.
I listened to The Age of Miracles by Karen Thompson Walker on audio, and I'm not sure why, but I didn't respond to this with as much enthusiasm as I thought I would. Parts of the story were great, the depiction of how people responded to the totally mysterious slowing down of the earth's rotation in particular. I liked how simply and naturally Walker describes what this was like. The integration of the sci-fi elements with a coming-of-age story was well-done as well. But the coming of age story itself seemed a little cliché. I didn't really like the teenage romance element.
Then there was Black Swan Green by David Mitchell, which I thought was fabulous. I suppose this is a fairly cliché coming-of-age story as well, but the writing was very, very good, which made up for it. Mitchell has a marvelous way with a sentence. It's a novel-in-stories, each chapter forming its own vignette in the life of the main character, a thirteen year-old boy who struggles with bullies and a stammer. Mitchell captures this character and the setting in which he lives very well.
For my mystery book group, I read Dorothy Hughes's In a Lonely Place, a book I chose after having heard good things about Hughes. It turned out to be a good choice, as the group liked her, and the discussion was lively. It's told in the first person from the perspective of the murderer, and the mood is unsettling and claustrophobic. We spent a lot of time trying to figure out the extent to which the narrator is unreliable and what exactly the other characters figured out and when. I like that sort of puzzle.
From the library, I got a copy of Jami Attenberg's The Middlesteins, a book I thought was very well done, a good, entertaining piece of literary fiction that made me feel a little dissatisfied with the state of literary fiction generally. I can't pinpoint anything wrong with the book, but I guess I'm in the mood for books that are more innovative or do something more exciting on the sentence level. It's a book about a family in Chicago and their struggles with a wife/mother who is seriously ill because of her weight. The descriptions of the family dynamics are good and if you're in the mood for a family drama, you might very well like it more than I did.
Then Kate Zambreno's book Heroines, which I liked with some reservations. It's partly literary criticism, history, and biography, and partly memoir. I enjoyed the combination of these things. Zambreno focuses on the "wives of modernism," writers such as Zelda Fitzgerald, Vivien Eliot, and others who were kept from writing or whose writing was dismissed and ignored because of their gender. Zambreno analyzes the language used to belittle these writers and the ideas about women and creativity that still influence us today. All this I liked. I just wished the book had a clearer organizational structure, as it felt repetitious and too long.
Finally, I just finished Meghan O'Rourke's memoir about her mother's death, The Long Goodbye. This is a book that grew on me as I read; at first it seemed to be a fairly unremarkable story about illness that I wished had more reflection rather than straightforward narrative. The reflective elements of the book became more important as it went on, however, and the second half or so has a lot of interesting insights into grief and mourning.


At its Executive Committee meeting on 10 March, the ETUC, in coordination with the ETF, adopted a declaration on low fares airlines in which the Confederation expresses its concern these companies undermine collective bargaining. The declaration refers to recent disputes in Norway and Denmark demonstrating those companies' opposition and actions to undermine the fundamental right of workers to negotiate and agree on working conditions and wages with their employer. The European Trade Union Confederation calls for imminent better and adapted regulation of the European airline sector to stop the race to the bottom.


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13 Dec | Monday
The Central Bank of the UAE (CBUAE) hosted the 39th meeting of the Islamic Financial Services Board (IFSB) Council and Capacity Building for Market Players on 9 December 2021 in Abu Dhabi, and co-organized the 13th IFSB Public Lecture, aiming to promote the development of a prudent, resilient, sustainable and transparent Shari'ah-compliant global financial services industry.
The 39th IFSB Council considered plans to transform the IFSB's strategic directions and policies to continue to meet industry's expectation and achieve the desired impact in IFSB member jurisdictions and globally. The Council recognised the importance of IFSB's focus on addressing emerging risks and vulnerabilities associated with evolving global developments related to environmental, social and governance (ESG), sustainability, climate change, impacts of accelerated digitisation, cyber-resilience, the elimination of LIBOR and central bank digital currencies (CBDCs). The Council noted the desired, potentially-significant role for Islamic Finance in providing Shari'ah-compliant solutions beyond the Islamic world, due to its pure principles and ethical investment aspects that addressed the ESG goals of global investors.
H.E. Khaled Mohamed Balama, Governor of the CBUAE and Chairman of the IFSB Council for 2021, commented that "boosting the role of Shari'ah-compliant financial solutions was an important part of the UAE's future financial plans. The CBUAE remained committed to the recouperation of global financial systems, in partnership with IFSB, its members and counterparts, as banking sector liquidity returned to pre-COVID levels."
H.E. added: "The IFSB would need to continue to be prudent and to implement fiscal measures to stimulate demand and boost economic well-being of Islamic Finance in member jurisdictions. The Islamic financial services industry would continue to be instrumental in any economic recovery plan or initiative". The Chairman urged the IFSB to seize the opportunity "to supercharge its ambitions, share knowledge to standardise Shari'ah financial principles and to deliver continuous, synchronised success across the IFSB's mandate with greater member engagement".
The IFSB Secretary-General, Dr Bello Lawal Danbatta appreciated the directions, contributions and outcomes of the 39th IFSB Council Meeting. He commended the Council for its unwavering support, commitment and outstanding guidance for the Secretariat. He added " as we now turn our minds to greater impact through implementation of IFSB standards, the IFSB will continue to count on the enhanced engagement of its members, their goodwill and their support. As always, we are grateful for your guidance on the way forward. Change takes courage. But, as our agendas and new Strategic Performance Plan (SPP) 2022-2024 make clear, business as usual will not mean that things will remain the same in the IFSB. Business as usual to us guarantees a future of economic and financial stability, policy and human capital transformation and many other initiatives. We hope that we can work together to chart a more positive course for the IFSB and the global Islamic Finance industry."
The 39th meeting of the IFSB Council, hosted by Central Bank of the United Arab Emirates was held on 9 December 2021 in Abu Dhabi, United Arab Emirates and via online. The Council Meeting was chaired by H.E. Khaled Mohamed Balama, Governor of Central Bank of the United Arab Emirates and Chairman of the IFSB for 2021, attended by the Central Bank Governors, Deputy Governors, Heads of regulatory and supervisory authorities, senior representatives from among the Council and Full members of the IFSB, representing 22 countries.


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Classic Car of the day: Standing out because…
Classic Car of the day: Standing out because it's ordinary
Mark Aldrich and his 1959 Nash Metropolitan 1500 Convertible. (James Raia — Herald Correspondent)
By James Raia | newsroom@montereyherald.com |
PUBLISHED: August 17, 2019 at 10:21 am | UPDATED: August 17, 2019 at 10:22 am
CARMEL — Vintage Ferraris and Porsches are fine and define precision performance. Mark Aldrich appreciates those iconic cars as much as anyone. But the Aliso Viejo lawyer's preferred vehicles are far more unheralded.
When high-end machines arrive at car shows with their distinctive growls, massive engines or flawless designs as mechanical works of art, Aldrich makes his entrance in a 1959 Nash Metropolitan 1500 Convertible.
Aldrich and a friend were relaxing in lawn chairs in the shade earlier this week, a few feet from the vehicle at the Concours on the Avenue. Commotion reigned nearby on Ocean Avenue, the main artery showcase for the 12th annual show, a mainstay of Classic Car Week.
Surrounded by muscle cars, a massive military vehicle and monstrous sedans replete with protruding body parts and lots of chrome, the 60-year-old little machine may have looked out of place. But it wasn't, and it never is in other automotive events.
"The car gets a lot of attention because it is unusual," said Aldrich, from his more serene location on Dolores Avenue. "We live near Laguna Beach where a Porsche doesn't get a second look. This car goes down the street and people look. My saying is 'Children laugh and dogs bark.' "
Aldrich bought the Metropolitan 33 years ago. One of his adult twin daughters drove it to high school. The other daughter didn't wait to drive it.
"My daughter called me once and said, 'Dad, someone stole the Metropolitan. We can't find it,' " Aldrich said. "I told her to look around."
Because of its petiteness and weight of only 1,800 pounds, members of the football team used to move the car around campus. In one instance, it was deposited on the 50-yard-line of the football field.
The little machine provides transportation as well as can be expected from a car with four cylinders, 52 horsepower, a top speed of 78 mph and a "three-on-the-tree" transmission. It runs on premium gas, and Aldrich adds a lead substitute to prevent the engine from running too hot.
For events like the Concours on the Avenue, Aldrich tows the Metropolitan with his Mercedes-Benz or with a trailer. He's taken the vehicle to car shows in Idaho, Arizona, Nevada and to several locales in California. He also brought the vehicle to the Concours on the Avenue in 2015. This year, it was entered in the American Family Favorites competition.
Aldrich's fondness for Nash, Rambler and other vehicles from American Motors Corporation dates to his childhood. His family owned a Rambler station wagon in the 1950s. In addition to his Metropolitan, Aldrich has also own Hornets and Gremlins made by AMC.
"One of my earliest memories was of my dad was when had a 1950 Nash station wagon towed into the backyard for me play in," Aldrich said. "It didn't have a motor or anything."
One prevailing theme during Classic Car Week is the value of the varied vehicles sold during the many auctions on the Peninsula.
Aldrich didn't discuss his vehicle's purchase price or the amount he spent to have it restored. It cost about $1,650 new and several vintage car online sites have 1959 Metropolitans for sale ranging from $9,800 to $24,000.
About 95,000 Metropolitans were made in England for Nash during their manufacturing tenure from October 1953 to April 1961. Slightly more than 22,000 sold new in 1959, the carmaker's best year.
Through the years, Aldrich has received several unsolicited offers for his car. He also tried to sell it once, but family members intervened.
"We just have a ball with it," said Aldrich.
James Raia
Stephen Curry broadcasts Warriors game: 'My plus-minus as a sideline reporter is phenomenal'


The Glynn-Johnson HL6 Push/Pull latch offers versatility and durability for commercial and institutional settings.
Sleek design has short throw angle, allowing the door to be opened with an easy push or pull action, even when hands are not free.
Push/Pull latch for 1-3/4" doors, (Including fire doors).
U.L. Listed for all labeled metal or wood doors.
No fusible link to fail.
Six mounting positions: Handles down, Handles up, One handle up, One handle down (either push or pull), Handles point away from latch, Handles point toward latch.
Function The short throw angle allows an easy push or pull action to open the door even when hands are not free.
Strike Plates ASA - 4-7/8"


Bend the Arc is a proud member of the Defund Hate coalition, led by Detention Watch Network, United We Dream, and other partners fighting for immigrant rights. So far, the coalition has successfully blocked over $5 billion in funding for border militarization and immigrant detention.
Our goal right now? Increasing the number of Congressmembers committed to defunding hate, educating our Jewish communities, growing public momentum, and combating the normalization of white supremacy.
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text:
Brestača är en ort i Kroatien.   Den ligger i länet Moslavina, i den östra delen av landet,  km sydost om huvudstaden Zagreb. Brestača ligger  meter över havet och antalet invånare är .

Terrängen runt Brestača är huvudsakligen platt, men österut är den kuperad. Den högsta punkten i närheten är  meter över havet,  km norr om Brestača. Runt Brestača är det glesbefolkat, med  invånare per kvadratkilometer. Närmaste större samhälle är Novska,  km sydost om Brestača. I omgivningarna runt Brestača växer i huvudsak lövfällande lövskog.

Trakten ingår i den hemiboreala klimatzonen. Årsmedeltemperaturen i trakten är  °C. Den varmaste månaden är juli, då medeltemperaturen är  °C, och den kallaste är december, med  °C. Genomsnittlig årsnederbörd är  millimeter. Den regnigaste månaden är september, med i genomsnitt  mm nederbörd, och den torraste är mars, med  mm nederbörd.

Kommentarer

Källor

Orter i Moslavina

redpajama_set_name: RedPajamaWikipedia


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Health Alert US
How To Relieve Arthritis, Joint Pain, and Inflammation Without A Prescription
'Winter blues' or something more? When to seek help for Seasonal…
Deaths related to binge-drinking on the rise, new studies show
Medical centers fight to keep liver transplants close to home, after…
Baby born to coronavirus patient tests positive for illness, Chinese state…
Home Health Alert US Baby born to coronavirus patient tests positive for illness, Chinese state media...
Baby born to coronavirus patient tests positive for illness, Chinese state media says
FILE - In this Wednesday, Jan. 22, 2020, file photo, a staff member moves bio-waste containers past the entrance of the Wuhan Medical Treatment Center in Wuhan, China, where some people infected with a new virus are being treated. The new virus comes from a large family of coronaviruses, some causing nothing worse than a cold. Others named SARS and MERS have killed hundreds in separate outbreaks. (AP Photo/Dake Kang, File)
A baby born to a woman with the coronavirus tested positive for the illness 30 hours after birth, raising the possibility that pregnant women may be able to pass the virus onto their unborn children, Chinese doctors said.
The baby, who was born Feb. 2, is being treated at Wuhan Children Hospital and is in stable condition with no fever or cough, but had experienced shortness of breath. Chest X-rays showed signs of infection and liver abnormalities, Reuters reported.
"This reminds us to pay attention to mother-to-child being a possible route of coronavirus transmission," Zeng Lingkong, chief physician of Wuhan Children's Hospital's neonatal medicine department told state broadcaster CCTV, according to Reuters.
Lingkong said a second case involving an infant who was born on Jan. 13 and later diagnosed with the virus on Jan. 29 is under investigation. It was not clear if the child's mother, who was diagnosed with coronavirus after the child's birth, or the family's nanny, who was also diagnosed with the illness, passed it on to the baby.
The illness has sickened over 24,000 people worldwide and killed at least 490. Wuhan is considered the virus epicenter, and 99 percent of the cases have occurred in mainland China. Two deaths have been reported outside the country, including one in Hong Kong and another in the Philippines.
Lingkong said the two cases confirm that infants are also at risk of contracting the virus. Earlier this week, photos of a medical team at Harbin Sixth People's Hospital in Heilongjiang who helped deliver the baby of a coronavirus patient emerged as staffers posed in their hazmat gear alongside the newborn.
The woman, who was not identified, went into labor on Jan. 30 after being diagnosed with the virus. She reportedly had close contact with relatives who had recently returned from Wuhan before her diagnosis. Doctors delivered her child via caesarian section and said that the newborn was born healthy and had so far tested negative for the virus.
It is known that the coronavirus may spread through respiratory droplets, but a new report suggests that it may also transmit through the digestive tract, specifically the fecal-oral route.
Scientists from the Renmin Hospital of Wuhan University and the Wuhan Institute of Virology of the Chinese Academy of Science discovered "virus genetic material" in stool samples and rectal swabs taken from patients, according to Chinese state media.
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Cosmic Dawn: Astronomers Find Fingerprints of Universe's First Stars
An artist's illustration of what the first stars in the universe may have looked like. (Image credit: N.R. Fuller, National Science Foundation)
The cosmic dark ages lasted no more than 180 million years.
Astronomers have picked up a long-sought signal from some of the universe's first stars, determining that these pioneers were burning bright by just 180 million years after the Big Bang.
Scientists had long suspected that dawn broke over the cosmos that long ago; theorists' models predict as much. But researchers had never had the evidence to back it up until now. Before this new study, the oldest stars ever seen dated to about 400 million years after the Big Bang. [The Universe: Big Bang to Now in 10 Easy Steps]
"This pushes our knowledge of when and how stars formed to earlier times in the universe," said study lead author Judd Bowman, an astronomer at Arizona State University's School of Earth and Space Exploration.
These very ancient stars were trailblazers. Though they coalesced from primordial hydrogen and helium, they set in motion a continuing process of star birth and death that ended up, over the eons, seeding the universe with heavy elements — the stuff that rocky planets like Earth are made of.
"If you look at our cosmic origins," Bowman told Space.com, "the bottom rung of that ladder is this process of the first objects forming and enriching the medium to make everything else possible."
In addition, the signal that Bowman and his team found was surprisingly strong. It was so strong, in fact, that it hints at a possible interaction between mysterious dark matter and the "normal" stuff that makes up the stars and you and me and everything else we can see in the universe.
Sifting through the noise
The further back in time you go, the harder it is to spot stars directly, using instruments such as NASA's Hubble Space Telescope. For starters, there are fewer and fewer stars to find. And until about 500 million years after the Big Bang, the universe was suffused with neutral hydrogen atoms, which are good at blocking light. (Radiation from the first stars eventually split these atoms into their constituent protons and electrons, creating a more transparent ionized plasma, but this took a while.)
So, Bowman and his colleagues took an indirect route, searching for the fingerprints these early stars likely left on the cosmic background radiation (CMB) — the ancient light left over from the Big Bang. The stars' ultraviolet radiation, the idea goes, would excite hydrogen atoms into a different state, causing them to absorb CMB photons.
Theoretically, this dip in the CMB signal should be detectable. So, the team built, calibrated and tested a radio antenna the size of a kitchen table — a project they called Experiment to Detect the Global EoR (Epoch of Reionization) Signature (EDGES), which was funded by the U.S. National Science Foundation (NSF).
The EDGES ground-based radio spectrometer, at CSIRO's Murchison Radio-astronomy Observatory in Western Australia. (Image credit: CSIRO Australia)
Then, they set the equipment up at the Murchison Radio-astronomy Observatory (MRO) in Western Australia. The MRO is in an extraordinarily radio-quiet area maintained by the Commonwealth Scientific and Industrial Research Organization, Australia's national science agency.
The radio-quiet aspect of the site was key, because modeling work suggested that the signal Bowman and his colleagues were looking for overlapped with frequencies on the FM radio dial. And the researchers already had to contend with all of the Milky Way's booming background radio noise. [Stunning Photos of Our Milky Way Galaxy (Gallery)]
"There is a great technical challenge to making this detection," Peter Kurczynski, the NSF program director who oversaw funding for EDGES, said in a statement. "Sources of noise can be 10,000 times brighter than the signal. It's like being in the middle of a hurricane and trying to hear the flap of a hummingbird's wing."
But EDGES picked up that tiny flap, spotting a dip that's most intense at a frequency of about 78 megahertz. Hydrogen emits and absorbs radiation at a wavelength equivalent to 1,420 megahertz, so the signal EDGES detected had been "redshifted" — stretched to lower frequencies by the expansion of the universe. The extent of this redshift told the team when those CMB photons were absorbed: about 180 million years after the universe's birth.
Bowman and his team reported these results today (Feb. 28), in a study published online in the journal Nature (opens in new tab).
"These researchers with a small radio antenna in the desert have seen farther than the most powerful space telescopes, opening a new window on the early universe," Kurczynski said.
The EDGES signal petered out less than 100 million years later, probably because X-ray light emitted by supernovas, black holes and other objects had heated up the hydrogen atoms significantly by that point, Bowman said.
A timeline of the universe, updated to show when the first stars emerged (by 180 million years after the Big Bang). (Image credit: N.R. Fuller, National Science Foundation)
Dark matter involved?
The signal EDGES found was about twice as strong as the team expected. There are two possible explanations for this surprising intensity, Bowman said: Either the radio background was quite a bit stronger in those early days than scientists had thought, or the hydrogen gas was significantly cooler.
The study team leans toward the second possibility, because it's tough to imagine a process that would increase the radio background to the necessary levels, Bowman said. It's also tricky to figure out what may have cooled down the hydrogen, but there is a promising contender: dark matter, the mysterious stuff that makes up 85 percent of the material universe.
Dark matter neither absorbs nor emits light, making it impossible to see directly (hence the name). Astronomers have inferred the substance's existence from its gravitational effects on "normal" matter, but they don't know what dark matter actually is. Most researchers think it's made up of as-yet-undiscovered particles, hypothesized specks such as axions or weakly interacting massive particles.
In a separate study (opens in new tab) in the same issue of Nature, astrophysicist Rennan Barkana, of Tel Aviv University in Israel, suggested that cold dark matter may have sucked away energy from the hydrogen gas, cooling it down. If this happened, "the dark-matter particle is no heavier than several proton masses, well below the commonly predicted mass of weakly interacting massive particles," Barkana wrote in his study.
If Barkana is right, Bowman and his team have gotten a look at some exotic physics and uncovered an important clue about the nature of dark matter. [Gallery: Dark Matter Throughout the Universe]
"We've been looking for so long for anything that can tell us more about what dark matter might be," Bowman said. "If this indeed gets borne out and continues to be confirmed — that the detection is real, and Rennan's hypothesis is real [and] is the best explanation — then this might well be the first key to advancing our knowledge of what dark matter really is."
Speaking of confirming the detection — that's the immediate next step in this line of early universe research, Bowman said. He and his team spent about two years validating their find, ruling out all possible alternative explanations. But for the discovery to be rock-solid, another research group needs to spot the signal as well.
If that happens, astronomers can mine the signal for more information, Bowman said. After all, now they know where to find it.
For example, further study by sensitive radio-telescope arrays should reveal more about the nonstandard physics hinted at by the signal and more about the properties of the universe's first stars, he said.
"Also, we would expect that we can eventually start to discern when the very first stars transitioned into the second-generation and later stars that were built out of gas that had heavier elements in it," Bowman said. "I think all of that then gets tied into the origin and formation of galaxies as a whole."
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