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Tadpoles of the Magnificent Tree Frog Litoria splendida produce host defence peptides early in their development and well before metamorphosis. Peptides were identified and characterized using high performance liquid chromatography and electrospray mass spectrometry. No host defence peptides were identified in the eggs. The neuropeptide caerulein was detected 10 d after egg deposition, and the antibiotic peptides caerin 1.1, caerin 1.6 and caerin 3.1 first appeared at 14 d. The concentration of peptides increases with the onset of metamorphosis at 84 d, when the host-defence peptide profile is the same as that of the adult. [ABSTRACT FROM AUTHOR]

With alarming frequency, significant collections in natural history museums have been destroyed or damaged through insurrections, cyclones, wars, fires, floods, or earthquakes particularly in the nineteenth century but continuing into the twentieth century with World War II bombings, fires, and earthquakes being the primary causes of loss in fifty-seven institutions across thirty countries. We review the loss or damage of museum collections globally, and their varied causes. We emphasize the benefits of dispersal of a sample of paratypes across institutions as an essential feature of taxonomic practice. We argue that museums do not own type material but are acting as perpetual custodians of type material on behalf of science and society in general, and that museums, therefore, have an obligation to minimize the risk to their collections. The significance of the loss of type material would be ameliorated if, when there are numerous paratypes or syntypes, members of a type series were distributed among several institutions. This is currently common practice but historically this was not always the case and might not be possible if only a single holotype is available. We also reaffirm the need for scientists around the globe to develop specific protocols to protect collections of biological and cultural materials from loss or damage from natural and human-created disasters now and into the future. We comment on recent moves to modify the Zoological Code of Nomenclature to allow the use of images as "type" material when describing new species with the image serving as a substitute for "physical" specimens deposited in museum collections. Although our focus is on herpetological collections, our particular interest and area of expertise, our observations apply broadly to all collections, including those of animals, plants, and anthropological or ethnographic material.

Advance directives can help guide care in the ICU. As a healthcare quality improvement initiative, we sought to increase the percentage of patients with a healthcare power of attorney and/or practitioner orders for life-sustaining treatment in our ICU and to increase medical resident experience with advance directives through routine screening and documentation of advance directives in the ICU., Design: Prospective analysis., Setting: Urban U.S. community teaching hospital., Patients: All patients admitted to the ICU from September 2018 to February 2019., Interventions: Internal medicine residents in the ICU received a lecture about advance directives and instructions to screen their patients for advance directives. For willing and decisional patients, residents facilitated the creation of a healthcare power of attorney and/or practitioner orders for life-sustaining treatment. Residents were anonymously surveyed at the beginning and end of the ICU rotation about their experience and level of comfort with healthcare power of attorney and practitioner orders for life-sustaining treatment completion., Measurements and Main Results: Three-hundred seventy-five patients were admitted to the ICU during the study period. Healthcare power of attorney documents were generated by 34% of all ICU patients without a prior healthcare power of attorney, increased from a baseline rate of 10% ( p < 0.001). The number of practitioner orders for life-sustaining treatment documents for patients with code status of "no cardiopulmonary resuscitation" did not increase significantly. The percentage of residents who facilitated completion of a healthcare power of attorney document increased significantly from 56% to 100% ( p < 0.001), whereas their practitioner orders for life-sustaining treatment experience did not change significantly by the end of their ICU rotation. On a Likert scale of 0-10, mean resident comfort increased significantly both with healthcare power of attorney documentation, rising from 6.14 to 8.84 ( p = 0.005) and with practitioner orders for life-sustaining treatment form completion, increasing from 6.00 to 7.84 ( p = 0.008)., Conclusions: Training ICU medical residents to routinely screen for and facilitate completion of advance directives significantly increased the percentage of ICU patients with a healthcare power of attorney and significantly improved medical resident comfort with healthcare power of attorney and practitioner orders for life-sustaining treatment form completion., (Copyright © 2021 The Authors. Published by Wolters Kluwer Health, Inc. on behalf of the Society of Critical Care Medicine.)

The major mode of transmission of the human immunodeficiency virus (HIV) is by sexual intercourse. In the effort to halt the spread of HIV, one measure that holds great promise is the development of effective microbicides that can prevent transmission. Previously we showed that several amphibian antimicrobial peptides (AMPs) completely inhibit HIV infection of T cells while maintaining good viability of the T cell targets. These peptides also inhibited the transfer of HIV by dendritic cells (DCs) to T cells when added up to 8h after virus exposure. Here we report on the anti-HIV activity of 18 additional structurally related caerin 1 family peptides in comparison with our previous best candidate caerin 1.9. Nine peptides were equally effective or more effective in the inhibition of T cell infection and disruption of the HIV envelope as caerin 1.9. Of those nine peptides, three peptides (caerin 1.2, caerin 1.10, and caerin 1.20) exhibited excellent inhibition of HIV infectivity at low concentrations (12-25μM) and limited toxicity against target T cells and endocervical epithelial cells. There was a direct correlation between the effectiveness of the peptides in disruption of the viral envelope and their capacity to inhibit infection. Thus, several additional caerin 1 family peptides inhibit HIV infection have limited toxicity for vaginal epithelial cells, and would be good candidates for inclusion in microbicide formulations., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The dorsal adhesive secretion of the frog Notaden bennetti and the prey-capture "slime" ejected by Euperipatoides sp. velvet worms look and handle similarly. Both consist largely of protein (55-60% of dry weight), which provides the structural scaffold. The major protein of the onychophoran glue (Er&#95;P1 for Euperipatoides rowelli) and the dominant frog glue protein (Nb-1R) are both very large (260-500 kDa), and both give oddly "turbulent" electrophoresis bands. Both major proteins, which are rich in Gly (16-17 mol%) and Pro (7-12 mol%) and contain 4-hydroxyproline (Hyp, 4 mol%), have the composition of intrinsically unstructured proteins. Their propensities for elastomeric or amyloid structures are discussed in light of Er&#95;P1's large content of intrinsically disordered long tandem repeats. The low carbohydrate content of both glues is consistent with conventional protein glycosylation, which in the N. bennetti adhesive was explored by 2D PAGE. The N-linked sugars of Nb-1R appear to prevent inappropriate self-aggregation. Some peptide sequences from Nb-1R are presented. Overall, there are enough similarities between the frog and the velvet worm glues to suspect that they employ related mechanisms for setting and adhesion. A common paradigm is proposed for amphibian and onychophoran adhesives, which, if correct, points to convergent evolution., (Copyright © 2013 Elsevier Inc. All rights reserved.)

A previous review summarized research prior to 2004 carried out on the bioactive host-defense peptides contained in the skin secretions of Australian anurans (frogs and toads). This review covers the extension of that research from 2004 to 2012, and includes membrane-active peptides (including antibacterial, anticancer, antifungal and antiviral peptides) together with the mechanisms by which these peptides interact with model membranes, peptides that may be classified as "neuropeptides" (including smooth muscle active peptides, opioids and immunomodulators) and peptides which inhibit the formation of nitric oxide from neuronal nitric oxide synthase. The review discusses the outcome of cDNA sequencing of signal-spacer-active peptides from an evolutionary viewpoint, and also lists those peptides for which activities have not been found to this time., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The Kyn-containing peptide FP-Kyn-L(NH(2)) is an unusual minor component of the skin peptide profile of the Australian red tree frog Litoria rubella collected from an area within a 20 kilometre radius of Alice Springs in central Australia. The structure was determined by electrospray mass spectrometry and synthesis. The major component of the skin secretion is the analogous tryptophyllin peptide FPWL(NH(2)). Both peptides show opioid activity at 10(-7) M, and are likely to act via the μ opioid receptor., (Copyright © 2011 John Wiley & Sons, Ltd.)

The submandibular musculature of 37 species of the five currently recognized genera of the subfamily Phyllomedusinae (Anura: Hylidae) is described; observations are made on the variation and ontogeny of these muscles. Supplementary apical elements of the m. intermandibularis occur in all phyllomedusines studied, in addition to the supplementary posterolateral elements previously reported. Our observations are discussed in the context of 1) the proposed homology between supplementary apical and posterolateral elements; 2) the homology with the apical elements reported for Pelodryadinae (sister taxon of Phyllomedusinae); and 3) the implications for our understanding of the relationships between Phyllomedusinae and Pelodryadinae. Anatomical differences between the apical and posterolateral elements and their co-occurrence in phyllomedusines indicate that these supplementary elements are not homologous. Despite differences between phyllomedusines and pelodryadines in the adhesion of supplementary fibers to the principal element of the m. intermandibularis and the occurrence of a broad aponeurosis or a medial raphe, the extensive morphological and developmental resemblances of the apical elements indicate that these structures are homologous, and that the presence of apical elements is a synapomorphy of Phyllomedusinae + Pelodryadinae., (Copyright © 2011 Wiley-Liss, Inc.)

Host defense peptides of 35 species of Australian frogs from the hylids Cyclorana and Litoria, and the myobatrachids Crinia, Limnodynastes and Uperoleia have been identified. The biological activities of the majority of these peptides have been determined and include hormones, neuropeptides, opioids, immunomodulators, membrane active peptides [including antimicrobial, anticancer, antiviral (enveloped viruses like HIV and Herpes) and antifungal peptides], neuronal nitric oxide synthase inhibitors, pheromones and individual peptides with other specific activities. The host defense peptide skin profile can be diagnostic at both the species and higher taxonomic levels; for example, species of Crinia, Litoria and Uperoleia each produce quite different types of peptides. Species of Cyclorana and Limnodynastes are more difficult to characterize by skin peptides alone: species of both genera produce similar peptides with no apparent activity. The skin peptide profiles of frogs from the genera Crinia, Litoria and Uperoleia may be used together with morphological and cognate methods, to differentiate between sub-species and even different population clusters of the same species. Nucleotide sequencing of cDNAs of precursors (pre-pro peptides) of bioactive peptides from the skin glands of various species of the genus Litoria show that the majority of these peptides originated from a single ancestor gene before the break away of Australia from Gondwana. The exceptions are the caerulein neuropeptides {e.g. caerulein [pEQDY(SO(3)H)TGWMDF(NH(2))]} which have a different origin to that of other Litoria peptides. Disulfide containing peptides from skin glands of species of Crinia show a different evolutionary route to peptides from species of Litoria., (Copyright © 2010 Elsevier Inc. All rights reserved.)

When provoked, Notaden bennetti frogs secrete a proteinaceous exudate, which rapidly forms a tacky and elastic glue. This material has potential in biomedical applications. Cultured cells attached and proliferated well on glue-coated tissue culture polystyrene, but migrated somewhat slower than on uncoated surfaces. In organ culture, dissolved glue successfully adhered collagen-coated perfluoropolyether lenses to debrided bovine corneas and supported epithelial regrowth. Small pellets of glue implanted subcutaneously into mice were resorbed by surrounding tissues, and all of the animals made a full recovery. An initial but transient skin necrosis at the implant site was probably caused by some of the potentially toxic metabolites present in the frog secretion; these include sterols and carotenoids, as well as fatty alcohols, aldehydes, ketones, acids, and aromatic compounds. Removal of the carotenoid pigments did not significantly alter the glue's material properties. In contrast, peroxidase treatment of dissolved glue introduced unnatural crosslinks between molecules of the major protein (Nb-1R) and resulted in the formation of a soft hydrogel, which was very different to the original material., (Copyright 2009 Wiley Periodicals, Inc.)

Positive and negative ion electrospray mass spectrometry together with Edman sequencing (when appropriate) has been used to sequence the host-defence peptides secreted from skin glands of the tree frog Litoria peronii. The peptide profiles are different in winter and summer. In winter, the frog produces small amounts of the known caerin 1.1 [GLLSVLGSVAKHVLPHVVPVIAEHL-NH(2)] (a wide-spectrum antibiotic) and caerin 2.1 [GLVSSIGRALGGLLADVVKSKQPA-OH], a narrow-spectrum antibiotic and an inhibitor of neuronal nitric oxide synthase. The major peptides produced throughout the year are the pGlu-containing peroniins 1.1 to 1.5 (e.g. peroniin 1.1 [pEPWLPFG-NH(2)], a smooth muscle contractor from 10(-7) M), and caerulein [pEQDY(SO(3)H)TGWMDF-NH(2)], a known and potent smooth muscle contractor from 10(-10) M. There are also some precursors to the peroniin 1 peptides, only detected in the skin secretion in summer, which are inactive and appear to be all (or part) of the spacer peroniin 1 peptides, e.g. peroniin 1.1b [SEEEKRQPWLPFG-NH(2)]. There are three members of the Litoria peronii Group of tree frogs classified in Australia, namely, L. peronii, L. rothii and L.tyleri. A comparison of the skin peptide profiles of L. peronii with those reported previously for L. rothii suggests that either these two species of tree frog are not as closely related as determined previously on morphological grounds, or that skin peptide divergence in tree frogs of this Group is more extensive than in others that have been studied., (Copyright (c) 2009 John Wiley & Sons, Ltd.)

Summary: Rotator cuff tendons are typically reattached to the proximal humerus using transosseous sutures or suture anchors. Their primary mode of failure is at the tendon-bone interface. We investigated the addition of a novel adhesive secreted from a species of Australian frog (Notaden bennetti) to different methods of rotator cuff repair. We hypothesized that the addition of frog glue would increase the strength of the repaired rotator cuff construct. Three techniques were used to repair 42 fresh frozen sheep infraspinatus tendons with a mattress stitch configuration: transosseous sutures; 2 traditional metallic suture anchors with 1 suture per anchor, and 2 knotless metallic anchors with 1 suture per anchor. In each group, 7 shoulders were repaired with the addition of frog glue to the infraspinatus "footprint," whereas 7 were used as control with no adhesive. Failure occurred in all constructs at the tendon-bone-suture interface. Repair with suture anchors was stronger than with sutures through bone (P < .05). Frog glue significantly increased the load to failure, total energy required for failure, and maximum energy at failure in all repair techniques (P < .01). A 2-fold increase occurred in load to failure of the 2 common anchor types (143 +/- 8 and 165 +/- 20 N). The load to failure for the transosseous repair (86 +/- 8 N) increased 1.7-fold. The addition of an adhesive to the tendon-bone-suture interface significantly enhances ultimate load and total energy required to failure in 3 types of rotator cuff repair. The unique properties of this frog glue (strong, flexible and sets in water) may ultimately lead to its use as an adjunct to rotator cuff repair in humans., Level of Evidence: Basic science study.

Many species of frogs of the genus Litoria secrete bioactive peptides from their skin glands. These peptides are normally host-defence compounds and may have one, or more of the following activities; smooth muscle contraction, analgesic, antimicrobial, antiviral, lymphocyte proliferator (immunomodulator) and neuronal nitric oxide synthase (nNOS) inactivation. Two frog species of the Litoria rubella Group that have been studied before, namely, Litoria electrica and Litoria rubella, are different from other species of the genus Litoria in that they produce small peptides that show neither membrane, lymphocyte nor nNOS activity. In this study we have used electrospray mass spectrometry together with Edman sequencing to identify eight skin peptides of the third member of this Group, Litoria dentata: surprisingly, none of these peptides show activity in our biological screening program. However, two major peptides (FPWL-NH(2) and FPWP-NH(2)) from L. electrica and L. rubella are opioids at the micromolar concentration.

The skin secretions of Crinia signifera, C. riparia and C. deserticola contain bioactive disulfide-containing peptides. Signiferin 1 (RLCIPYIIPC-OH) from C. signifera and C. deserticola) contracts smooth muscle at a concentration of 10(-9) M, and effects proliferation of lymphocytes at 10(-6) M. In contrast, riparin 1.1 (RLCIPVIFC-OH) and riparin 1.2 (FLPPCAYKGTC-OH) from C. riparia show lymphocyte activity but do not contract smooth muscle. The lymphocyte and smooth muscle activities involve CCK2R. 3D structures of signiferin 1 and riparin 1.1 have been established using 2D NMR methods: these studies show significant differences in the shapes of the disulfide rings and with the orientations of the N-terminal residues. cDNA cloning establishes that the pre sections of the precursor pre-pro-riparin 1.4-1.6 peptides are different from the conserved pre regions of disulfide-containing antimicrobial peptides from species of the genus Rana found in the northern hemisphere and caerin antimicrobial peptides isolated from Australian tree frogs of the genus Litoria. This suggests that (i) either that riparins 1 have converged to similar structure and function to the ranid and hyloid prepropeptides which were lost initially from the myobatrachid lineage, or (ii) the prepropeptides in all three groups were derived from a single ancestral form that has remained relatively conserved in the hyloid and ranoid lineages but has undergone substantial divergent evolution in the myobatrachids.

The glandular skin secretion of the Eastern Dwarf Tree Frog Litoria fallax contains nine peptides named fallaxidins. The sequences of these peptides were elucidated using a combination of positive and negative electrospray mass spectrometry together with Edman sequencing. Among these peptides are: (i) fallaxidins 1.1 and 2.1 which have the sequences YFPIPI-NH2 and FWPFM-NH2. The activities of these peptides are unknown, but it has been shown that they are not smooth muscle active, opioids or antimicrobially active, nor do they effect proliferation of lymphocytes; (ii) two weakly active antibiotics, fallaxidins 3.1 and 3.2 (e.g. fallaxidin 3.1, GLLDLAKHVIGIASKL-NH2), and a moderately active antibiotic fallaxidin 4.1 (GLLSFLPKVIGVIGHLIHPPS-OH). Fallaxidin 4.1 has an unusual sequence for an antibiotic, containing three Pro residues together with a C-terminal CO2H group. cDNA cloning has confirmed the identity of the nine isolated peptides from L. fallax, together with five additional peptides not detected in the peptide profile. The pre-regions of the nine preprofallaxidins are conserved and similar to those of the caerin peptides from L. caerulea and L. splendida, suggesting that the fallaxidin and caerin peptides, although significantly different in sequence, originated from a common ancestor gene.

A dominant mechanism of cellular patterning in the growing fish retina is control of cell fate acquisition by negative feedback signals arising from differentiated cells. We tested the ability of a computational model of this pattern formation mechanism to simulate cellular patterns in regenerated goldfish retina. The model successfully simulated quantitative features of in vivo regenerated patterns, indicating that regenerating retina has access to and utilizes patterning mechanisms that are operational during normal growth. The atypical patterns of regenerated retina could arise in part from regenerative progenitors that, compared to normal growth progenitors, are less responsive to the feedback patterning signals.

Five healthy adult female first-generation hybrid tree frogs were produced by interspecific breeding of closely related tree frogs Litoria splendida and L. caerulea in a cage containing large numbers of males and females of both species. Phylogenetic analysis of mitochondrial DNA sequences established the female parent to be L. splendida. The peptide profile of the hybrid frogs included the neuropeptide caerulein, four antibiotics of the caerin 1 family and several neuronal nitric oxide synthase inhibitors of the caerin 1 and 2 classes of peptides. The skin secretions of the hybrids contained some peptides common to only one parent, some produced by both parental species, and four peptides expressed by the hybrids but not the parental species.

Visual search based on a conjunction of two features typically elicits reaction times that increase linearly as a function of the number of distractors, whereas search based on a single feature is essentially unaffected by set size. These and related findings have often been interpreted as evidence of a serial search stage that follows a parallel search stage. However, a wide range of studies has been showing a form of blending of these two processes. For example, when a spoken instruction identifies the conjunction target concurrently with the visual display, the effect of set size is significantly reduced, suggesting that incremental linguistic processing of the first feature adjective and then the second feature adjective may facilitate something approximating a parallel extraction of objects during search for the target. Here, we extend these results to a variety of experimental designs. First, we replicate the result with a mixed-trials design (ruling out potential strategies associated with the blocked design of the original study). Second, in a mixed-trials experiment, the order of adjective types in the spoken query varies randomly across conditions. In a third experiment, we extend the effect to a triple-conjunction search task. A fourth (control) experiment demonstrates that these effects are not due to an efficient odd-one-out search that ignores the linguistic input. This series of experiments, along with attractor-network simulations of the phenomena, provide further evidence toward understanding linguistically mediated influences in real-time visual search processing.

The search for novel insect repellents has been driven by health concerns over established synthetic compounds such as diethyl-m-toluamide (DEET). Given the diversity of compounds known from frog skin and records of mosquito bite and ectoparasite infestation, the presence of mosquito repellents in frogs seemed plausible. We investigated frog skin secretions to confirm the existence of mosquito repellent properties. Litoria caerulea secretions were assessed for mosquito repellency by topical application on mice. The secretions provided protection against host-seeking Culex annulirostris mosquitoes. Olfactometer tests using aqueous washes of skin secretions from L. caerulea and four other frog species were conducted to determine whether volatile components were responsible for repellency. Volatiles from Litoria rubella and Uperoleia mjobergi secretions were repellent to C. annulirostris, albeit not as repellent as a DEET control. The demonstration of endogenous insect repellents in amphibians is novel, and demonstrates that many aspects of frog chemical ecology remain unexplored.