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Abstract Circulating tumor DNA (ctDNA) is the cornerstone of liquid biopsy diagnostics, revealing clinically relevant genomic aberrations from blood of cancer patients. Genomic analysis of single circulating tumor cells (CTCs) could provide additional insights into intra-patient heterogeneity, but it requires whole-genome amplification (WGA) of DNA, which might introduce bias. Here, we describe a novel approach based on mass spectrometry for mutation detection from individual CTCs not requiring WGA and complex bioinformatics pipelines. After establishment of our protocol on tumor cell line-derived single cells, it was validated on CTCs of 33 metastatic melanoma patients and the mutations were compared to those obtained from tumor tissue and ctDNA. Although concordance with tumor tissue was superior for ctDNA over CTC analysis, a larger number of mutations were found within CTCs compared to ctDNA (p = 0.039), including mutations in melanoma driver genes, or those associated with resistance to therapy or metastasis. Thus, our results demonstrate proof-of-principle data that CTC analysis can provide clinically relevant genomic information that is not redundant to tumor tissue or ctDNA analysis.

We present a taxonomic review and systematic revision of the Eyelash Palm-Pitviper Bothriechis schlegelii (Berthold, 1846) based on examination of 400 museum specimens, a phylogeographic analysis of 818 locality records, and 80 individuals sampled for molecular characters. We find morphological and phylogenetic support for five new species of Bothriechis Peters, 1859, which we describe here based on their unique combination of molecular, meristic, hemipenial, and color pattern characteristics. They are: B. klebbai sp. nov., B. rasikusumorum sp. nov., B. khwargi sp. nov., B. rahimi sp. nov., and B. hussaini sp. nov. Finally, we revalidate the names B. nigroadspersus (Steindachner, 1870), B. nitidus (Günther, 1859), and B. torvus (Posada Arango, 1889a), and provide a redescription of B. schlegelii.

Aortic stenosis (AS) is a prevalent valvular disorder that poses a significant burden on healthcare systems due to its debilitating symptoms and high mortality rates if left untreated. Surgical aortic valve replacement (SAVR) and transcatheter aortic valve replacement (TAVR) are the primary interventions for severe AS, but perioperative complications such as bleeding remain a concern. Von Willebrand factor (VWF), a crucial player in hemostasis, is known to be altered in AS and may contribute to the hemostatic imbalance observed in these patients. This prospective study aimed to investigate the association between prosthetic valve type, size, and postprocedural VWF levels in patients undergoing aortic valve replacement (AVR) for severe AS. This study involved 39 consecutive patients diagnosed with severe AS who underwent SAVR or TAVR. By elucidating the VWF dynamics associated with different prosthetic valves, this study sought to provide valuable insights into personalized valve selection and perioperative management strategies.

Neacomys is a genus of small spiny or bristly sigmodontine rodents that are common components of mammalian faunas in multiple biomes on Central and South America. Recent studies on this group have demonstrated that there is cryptic diversity yet to be discovered within currently recognized species that have not received comprehensive revisions, as well as in areas that have been overlooked. Here we ratify this assertion by describing a new species previously misidentified as the Narrow-footed Spiny Mouse (Neacomys tenuipes) from the Chocó biogeographic region in northwestern Ecuador, Neacomys marci Brito & Tinoco, sp. nov. Distinctiveness of this entity is supported by the combination of the following morphological characters: small size (head-body length 65–85 mm); long tail (69–126% longer than head-body length); pale buff-colored but gray-based belly fur; white throat; hypothenar pad usually absent; long nasals; and a condylar process higher than the coronoid process. Likewise genetic distance analyses and phylogenetic reconstructions based on cytochrome-b (Cytb) sequence data indicate a clear divergence from typical populations of N. tenuipes, and a sister relationship between them. The results presented here increase the diversity of Neacomys to 24 species, placing it among the most diverse genera within the sigmodontine rodents.

Threadsnakes are known for their conserved external morphology and historically controversial systematics, challenging taxonomic, biogeographic and evolutionary researches in these fields. Recent morphological studies—mostly based on µCT data of the skull and lower jaw—have resolved systematic issues within the group, for instance leading to the description of new taxa or re-positioning little-known scolecophidian taxa in the tree of life. Herein we describe a new polymorphic species of the genus Siagonodon from Amazonia based on morphological (external, osteology and hemipenis) and molecular data, and provide the first hemipenial description for the genus. We also reassign Siagonodon acutirostris to the genus Trilepida based on osteological data in combination with molecular evidence. The new species described represents an evolutionary paradox for scolecophidians because the species displays a remarkable variation in the shape of the snout region that is otherwise always highly conserved in this clade. Finally, this study reinforces the importance of protected areas as essential in maintaining vertebrate populations, including those that are not yet formally described.

The diversity of the oryzomyine rat Mindomys (Cricetidae, Sigmodontinae, Oryzomyini), is doubled here with the description of a new species from the remote Cordillera de Kutukú (Ecuador). The novel form can be easily differentiated from Mindomys hammondi –type species of the genus– by a large set of anatomical traits including, among others, larger jugals, parietal “wings” extending to zygomatic roots, larger otic capsules, well-exposed petrosals, narrow zygomatic plates almost without upper free borders, foramen magnum caudally oriented, larger molars, and accessory root of first upper molar present. Until now, the records of Mindomys were restricted to western Andean foothills. The material from Kutukú highlights an Amazonian species and reinforces the valuable biological significance of isolated mountain ranges in eastern Ecuador. Since Mindomys shows some external traits classically related to arboreal life, here we present a brief reappraisal of this poorly explored topic. A partially neglected anatomical system in sigmodontine studies, the fore feet, encloses crucial information reflecting arboreality.

The amerophidian snake radiation is a Late Cretaceous superfamily that encompasses two families: Aniliidae, pipe snakes, and Tropidophiidae, dwarf boas. We describe a new dwarf boa snake species, from the Tropidophiidae family, from the cloud forest in northeastern Ecuador. Tropidophis cacuangoae sp. nov. can be diagnosed from its congeners based on external and osteological morphology. The new species inhabits eastern tropical piedmont and lower evergreen montane forests, in the Amazon Tropical Rainforest biome, and is likely to be an Ecuadorian endemic. We also discuss the relationships of the new species with South American tropidophiids and provide a key to the identification of mainland South American dwarf boas.

Immunosorbent turnip vein clearing virus (TVCV) particles displaying the IgG-binding domains D and E of Staphylococcus aureus protein A (PA) on every coat protein (CP) subunit (TVCVPA) were purified from plants via optimized and new protocols. The latter used polyethylene glycol (PEG) raw precipitates, from which virions were selectively re-solubilized in reverse PEG concentration gradients. This procedure improved the integrity of both TVCVPA and the wild-type subgroup 3 tobamovirus. TVCVPA could be loaded with more than 500 IgGs per virion, which mediated the immunocapture of fluorescent dyes, GFP, and active enzymes. Bi-enzyme ensembles of cooperating glucose oxidase and horseradish peroxidase were tethered together on the TVCVPA carriers via a single antibody type, with one enzyme conjugated chemically to its Fc region, and the other one bound as a target, yielding synthetic multi-enzyme complexes. In microtiter plates, the TVCVPA-displayed sugar-sensing system possessed a considerably increased reusability upon repeated testing, compared to the IgG-bound enzyme pair in the absence of the virus. A high coverage of the viral adapters was also achieved on Ta2O5 sensor chip surfaces coated with a polyelectrolyte interlayer, as a prerequisite for durable TVCVPA-assisted electrochemical biosensing via modularly IgG-assembled sensor enzymes.

We have discovered a spectacular new species of frog in the genus Hyloscirtus, belonging to the H. larinopygion species group. The adult female is characterized by a mostly black body with large bright red spots on the dorsal and ventral surface, extremities, and toe pads. The adult male is unknown. Small juveniles are characterized by a yellow body with variable black markings on the flanks; while one larger juvenile displayed irregular orange or yellow marks on a black background color, with light orange or yellow toe pads. Additional distinctive external morphological features such as cloacal ornamentation are described, and some osteological details are imaged and analyzed. The performed phylogeny places the new species as the sister to a clade consisting of ten taxa, all of which are part of the H. larinopygion group. We use genetic distances to fit the new species into a published time-calibrated phylogeny of this group; our analysis based on the published chronology suggests that the divergence of the new species from its known congeners pre-dates the Quaternary period. The new species is currently only known only from Cerro Mayordomo, in Fundación EcoMinga´s Machay Reserve, at 2,900 m in the eastern Andes of Tungurahua province, Ecuador, near the southern edge of Los Llanganates National Park, but its real distribution may be larger.

Apostolepis is a diverse genus of dipsadid snakes, currently comprising 34 species occurring in most cis-Andean South America. The taxonomy of the group is highly unstable. Upon discovering its type series, we redescribe the rare species A. ambiniger (Peters, 1869) and provide an account of its geographic distribution and morphological variation in pholidosis, osteology, and hemipenial characters. We also discuss some aspects of the taxonomy of Apostolepis.

Apostolepis albicollaris and A. cerradoensis are two Elapomorphini snake species, described within a short timespan, from the Cerrado of central Brazil. In their brief descriptions, these two species were diagnosed from congeners largely based on highly variable external morphological characters. Interestingly enough, A. cerradoensis has remained known based on a single specimen since its description. Here, we present a reanalysis of both type specimens, as well as a careful examination of a large series of specimens formerly assigned to these species, based on the comparison of internal and external morphology. We conclude that both species are synonymous, providing evidence for the recognition of A. cerradoensis as a junior synonym of A. albicollaris. Furthermore, an account of its updated diagnosis, morphological variation, geographic distribution, hemipenial morphology, phylogenetic relationships and an osteological description are also provided. We also discuss its conservation status, suggesting that the species is under threat and qualifies to be listed as Vulnerable (VU ab(iii)), considering its rarity, small geographic range, and persistent environmental threats.

The Andean cloud forests of Ecuador are home to several endemic mammals. Members of the Thomasomyini rodents are well represented in the Andes, with Thomasomys being the largest genus (47 species) of the subfamily Sigmodontinae. Within this tribe, however, there are genera that have escaped a taxonomic revision, and Chilomys Thomas, 1897, constitutes a paradigmatic example of these “forgotten” Andean cricetids. Described more than a century ago, current knowledge of this externally unmistakable montane rodent is very limited, and doubts persist as to whether or not it is monotypic. After several years of field efforts in Ecuador, a considerable quantity of specimens of Chilomys were collected from various localities representing both Andean chains. Based on an extensive genetic survey of the obtained material, we can demonstrate that what is currently treated as C. instans in Ecuador is a complex comprising at least five new species which are described in this paper. In addition, based on these noteworthy new evidence, we amend the generic diagnosis in detail, adding several key craniodental traits such as incisor procumbency and microdonty. These results indicate that Chilomys probably has a hidden additional diversity in large parts of the Colombian and Peruvian territories, inviting a necessary revision of the entire genus.

Abstract Functional studies giving insight into the biology of circulating tumor cells (CTCs) remain scarce due to the low frequency of CTCs and lack of appropriate models. Here, we describe the characterization of a novel CTC‐derived breast cancer cell line, designated CTC‐ITB‐01, established from a patient with metastatic estrogen receptor‐positive (ER+) breast cancer, resistant to endocrine therapy. CTC‐ITB‐01 remained ER+ in culture, and copy number alteration (CNA) profiling showed high concordance between CTC‐ITB‐01 and CTCs originally present in the patient with cancer at the time point of blood draw. RNA‐sequencing data indicate that CTC‐ITB‐01 has a predominantly epithelial expression signature. Primary tumor and metastasis formation in an intraductal PDX mouse model mirrored the clinical progression of ER+ breast cancer. Downstream ER signaling was constitutively active in CTC‐ITB‐01 independent of ligand availability, and the CDK4/6 inhibitor Palbociclib strongly inhibited CTC‐ITB‐01 growth. Thus, we established a functional model that opens a new avenue to study CTC biology.

A review of the species of the genus Mylossoma Eigenmann & Kennedy, 1903 found in the Ecuadorian Cis-Andean region was conducted. Two species were recognized: M. albiscopum (Cope, 1872) from the Putumayo, Napo, and Tigre river basins and M. aureum (Agassiz, 1829) from the Napo river basin. A map of geographical distribution and radiographs for both species are provided. Historical records of M. duriventre (Cuvier, 1818) were not confirmed in this geographical distribution.

Xanthylium derivatives are curcumin analogs showing photochromic properties. Similarly, to anthocyanins, they follow the same multistate network of chemical species that are reversibly interconverted by external stimuli. In the present work, two new asymmetric monocarbonyl analogues of curcumin, 4-(4-hydroxy-3-metoxybenzylidene)-1,2,3,4-tetrahydroxanthylium chloride (compound 3) and 4-(4-hydroxybenzylidene)-6-methoxy-1,2,3,4-tetrahydroxanthylium chloride (compound 4) were synthesized, and their photochromic and biological properties were investigated. The UV-Vis spectroscopy and the direct and reverse pH-jumps studies confirmed the halochromic properties and the existence of different molecular species. A network of chemical reactions of these species was proposed. Furthermore, the antiproliferative properties of both compounds were evaluated using P19 murine embryocarcinoma cells and compared with each other. The results demonstrate that both new xanthylium derivatives modify the progression through the cell cycle of P19 cells, which translates into a significant antiproliferative effect. The effect of the methoxy group position is discussed and several checkpoint proteins are advanced as putative targets.

We describe a new species of Neotropical spiny-lizard of the genus Echinosaura from the Imbabura and Carchi Provinces on the western slopes of the Andes in northwestern Ecuador. The new species mostly resembles E. horrida. However, it can be distinguished from all congeners by having keeled enlarged dorsal scales forming a paired vertebral row, two paravertebral series of short oblique rows of projecting scales, and a pair of spine-like scales on temporal and nuchal regions. We also provide a detailed description of the osteology of the skull and pectoral girdle of the new species and present a phylogenetic hypothesis for Echinosaura based on three mitochondrial genes (12S, 16S, ND4) and one nuclear gene (c-mos).

We describe a new species of terrestrial-breeding frog of the genus Noblella from the northwestern slopes of the Andes of Ecuador, in the province of Pichincha, Ecuador, and report a new locality for the recently described N. worleyae. We include a detailed description of the osteology of both species and discuss their phylogenetic relationships. The new species is differentiated from other species of Noblella by having discs of fingers rounded, without papillae; distal phalanges only slightly T-shaped; toes slightly expanded and rounded distally, without papillae; dorsum uniform brown with irregular suprainguinal dark brown marks; venter yellowish cream, ventral surfaces of legs and thighs reddish to brownish cream; and dark brown throat. The new locality for N. worleyae is located in Los Cedros Reserve, an area highly threatened by mining. We highlight the importance of protecting endemic species of small vertebrates in northwestern Ecuador.

The Andean cloud forests of western Colombia and Ecuador are home to several endemic mammals; members of the Oryzomyini, the largest Sigmodontinae tribe, are extensively represented in the region. However, our knowledge about this diversity is still incomplete, as evidenced by several new taxa that have been described in recent years. Extensive field work in two protected areas enclosing remnants of Chocó montane forest recovered a high diversity of small mammals. Among them, a medium-sized oryzomyine is here described as a new genus having at least three new species, two of them are named and diagnosed. Although externally similar to members of the genera Nephelomys and Tanyuromys, the new genus has a unique molar pattern within the tribe, being characterized by a noticeable degree of hypsodonty, simplification, lamination, and third molar compression. A phylogeny based on a combination of molecular markers, including nuclear and mitochondrial genes, and morphological data recovered the new genus as sister to Mindomys, and sequentially to Nephelomys. The new genus seems to be another example of a sigmodontine rodent unique to the Chocó biogeographic region. Its type species inhabits cloud forest between 1,600 and 2,300 m in northernmost Ecuador (Carchi Province); a second species is restricted to lower montane forest, 1,200 m, in northern Ecuador (Imbabura Province); a third putative species, here highlighted exclusively by molecular evidence from one immature specimen, is recorded in the montane forest of Reserva Otonga, northern Ecuador (Cotopaxi Province). Finally, the new genus is also recorded in southernmost Colombia (Nariño Department), probably represented there also by a new species. These species are spatially separated by deep river canyons through Andean forests, resulting in marked environmental discontinuities. Unfortunately, Colombian and Ecuadorian Pacific cloud forests are under rapid anthropic transformation. Although the populations of the type species are moderately abundant and occur in protected areas, the other two persist in threatened forest fragments.

We describe two new species of salamanders of the genus Oedipina, subgenus Oedopinola, from two localities on the northwestern foothills of Ecuador, at elevations between 921 and 1,067 m. These are the southernmost members of the genus. We examined different museum collections and we found just three specimens of Oedipina from Ecuador, obtained throughout the history of herpetological collections in the country. We identify two of the three specimens as new species, but refrain from assigning a specific identity to the third, pending further study. Oedipina villamizariorum sp. n. is a medium-sized member of the genus, with a narrow, relatively pointed head and blunt snout; dorsolaterally oriented eyes, moderate in size; and digits that are moderately long and having pointed tips. Oedipina ecuatoriana sp. n., somewhat larger, has a narrow head and broadly rounded snout; this new species differs from all known Oedipina by the distinctive presence of paired prefrontal bones and a reduced phalangeal formula: 0-0-1-0; 0-1-2-1-1. We provide detailed descriptions of the osteology of both new species. Finally, we present a phylogenetic hypothesis for the genus, including one of the two new species, based on partial sequences of mitochondrial DNA.

Morphology and distribution of the South American snake Chironius leucometapus (Serpentes: Colubridae). Chironius leucometapus was described more than 25 years ago as subspecies of C. fuscus from restricted area in the Department of Junín in central Peru, with no additional specimens reported since. Examination of 17 new specimens reveals that C. leucometapus is widespread along the Amazonian slopes of the Andes between central Peru and northern Ecuador. New morphological data including scalation, hemipenes, color in life, and the frst description of the skull of Chironius to be based on high-quality CT-scan images are presented. The number of species of Chironius from Ecuador is elevated to eight.

We describe a new species of Epictia based on eight specimens from Nicaragua collected and housed in the collection of the Natural History Museum Vienna for more than a century. The species differs from the congeners by the combination of external morphological characters: midtail scale rows 10; supralabials two, anterior one large and in broad contact with supraocular; infralabials four; subcaudals 14–19; middorsal scale rows 250–267; supraocular scales present; frontal scale distinct; striped dorsal color pattern with more or less triangular dark blotches on each scale; small white blotch in anterior part of dorsal surface of rostral present in five out of six specimens (two further specimens are lacking their heads); terminal spine and adjacent scales white. Eidonomic species separation from other Epictia spp. is also supported by a few qualitative and quantitative differences in vertebrae count and morphology. The new species is putatively assigned to the Epictia phenops species group based on external morphological characters and distribution.

We describe a new species of terrestrial frog of the genus Noblella from the eastern versants of the Ecuadorian Andes in the upper Pastaza watershed. Noblella naturetrekii sp. n. differs from its Ecuadorian congeners by the presence of a differentiated tympanic membrane and a weakly defined tympanic annulus, and eyelids with rounded tubercles. In addition, the new species is characterized by its blackish–dark brown ventral coloration scattered with little white dots and the absence of papillae at the tip of the fingers and toes. We provide a detailed description of the call and osteology of the new species. Finally, we present the most complete phylogeny of the genus, which confirms that Noblella is a non-monophyletic group.

Analysis of diet composition and morphological characters of the Peruvian lizard Microlophus stolzmanni (Squamata: Tropiduridae). Microlophus stolzmanni is a diurnal lizard that is endemic to the dry forest of northern Peru. Little is known about the ecology of the species and the composition of its diet never has been studied. The stomach contents and morphological features related to feeding behavior are analyzed herein. Microlophus stolzmanni is a semi-herbivorous food generalist that also consumes animal items. All age groups prefer sedentary prey for which M. stolzmanni forages actively. As the lizard matures, plant material becomes a more important part of the diet.

The rod-shaped nanoparticles of the widespread plant pathogen tobacco mosaic virus (TMV) have been a matter of intense debates and cutting-edge research for more than a hundred years. During the late 19th century, their behavior in filtration tests applied to the agent causing the 'plant mosaic disease' eventually led to the discrimination of viruses from bacteria. Thereafter, they promoted the development of biophysical cornerstone techniques such as electron microscopy and ultracentrifugation. Since the 1950s, the robust, helically arranged nucleoprotein complexes consisting of a single RNA and more than 2100 identical coat protein subunits have enabled molecular studies which have pioneered the understanding of viral replication and self-assembly, and elucidated major aspects of virus–host interplay, which can lead to agronomically relevant diseases. However, during the last decades, TMV has acquired a new reputation as a well-defined high-yield nanotemplate with multivalent protein surfaces, allowing for an ordered high-density presentation of multiple active molecules or synthetic compounds. Amino acid side chains exposed on the viral coat may be tailored genetically or biochemically to meet the demands for selective conjugation reactions, or to directly engineer novel functionality on TMV-derived nanosticks. The natural TMV size (length: 300 nm) in combination with functional ligands such as peptides, enzymes, dyes, drugs or inorganic materials is advantageous for applications ranging from biomedical imaging and therapy approaches over surface enlargement of battery electrodes to the immobilization of enzymes. TMV building blocks are also amenable to external control of in vitro assembly and re-organization into technically expedient new shapes or arrays, which bears a unique potential for the development of 'smart' functional 3D structures. Among those, materials designed for enzyme-based biodetection layouts, which are routinely applied, e.g., for monitoring blood sugar concentrations, might profit particularly from the presence of TMV rods: Their surfaces were recently shown to stabilize enzymatic activities upon repeated consecutive uses and over several weeks. This review gives the reader a ride through strikingly diverse achievements obtained with TMV-based particles, compares them to the progress with related viruses, and focuses on latest results revealing special advantages for enzyme-based biosensing formats, which might be of high interest for diagnostics employing 'systems-on-a-chip'.

Background: Solid epithelial tumors like breast cancer are the most frequent malignancy in women. Circulating tumor cells (CTCs) are frequently released from hypoxic areas into the blood, where CTCs face elevated oxygen concentrations. This reoxygenation might challenge the use of CTCs for liquid biopsy. Methods: We modeled this situation in vitro using the breast cancer cell lines—MCF-7, MDA-MB-468, MDA-MB-231—and the cell line BC-M1 established from DTCs in the bone marrow. Cells were cultured under hypoxia, followed by a reoxygenation pulse for 4 h, reflecting the circulation time of CTCs. Analyzed were gene products like EGFR, ErbB-2, EpCAM, PD-L1 on mRNA and protein level. Results: mRNAs of erbb2 or pdl1 and protein levels of PD-L1 displayed significant changes, whereas ErbB-2 protein levels remained constant. The strongest discrepancy between protein and mRNA levels under hypoxia was observed for EGFR, supporting the idea of cap-independent translation of egfr mRNA. Analyses of the phosphorylation of AKT, Erk 1/2, and Stat3 revealed strong alterations after reoxygenation. Conclusions: CTCs reaching secondary sites faster than reoxygenation could alter the mRNA and protein levels in the cells. CTC and DTC with high PD-L1 levels might become quiescent under hypoxia but were easily reactivated by reoxygenation.

Reproduction is described for four phyllodactylid geckos, Phyllopezus maranjonensis, Phyllodactylus delsolari, P. thompsoni and P. reissii from the upper Marañon Valley, Peru. Observations were made between March and May 2010 and voucher specimens, already housed in the Zoologisches Forschungsmuseum Alexander Koenig (ZFMK), were X-rayed to check for gravid females. Gravid female P. maranjonensis, P. delsolari and P. reissii contain two eggs, whereas P. thompsoni produces a single egg that is deposited in leaf litter or crevices of stone walls. It is likely that females of all of these species produce multiple clutches in one year; the reproductive period seems to be extended in at least two species. Furthermore, measurements of numerous specimens of all four species are presented herein, revealing that Phyllopezus maranjonensis is the largest species of New World geckos.

Neotropical monkey lizards (Polychrus) are arboreal lizards with compressed bodies, partially fused eyelids and strikingly long, whip-like tails. The eight currently recognized species occur in the lowlands of South and Central America. Based on the largest taxon and character sampling to date, we analyze three mitochondrial and one nuclear gene using Bayesian methods to (1) infer the phylogeny of Polychrus under both concatenated-tree and species-tree methods; (2) identify lineages that could represent putative undescribed species; and (3) estimate divergence times. Our species tree places P. acutirostris as the sister taxon to all other species of Polychrus. While the phylogenetic position of P. gutturosus and P. peruvianus is poorly resolved, P. marmoratus and P. femoralis are strongly supported as sister to P. liogaster and P. jacquelinae, respectively. Recognition of P. auduboni and P. marmoratus sensu stricto as distinct species indicates that the populations of "P. marmoratus" from the Amazon and the Atlantic coast in Brazil represent separate species. Similarly, populations of P. femoralis from the Tumbes region might belong to a cryptic undescribed species. Relative divergence times and published age estimates suggest that the orogeny of the Andes did not play a significant role in the early evolution of Polychrus.

Background/Aims: Clusterin (CLU), also known as Apolipoprotein J (ApoJ) is a highly glycosylated extracellular chaperone. In humans it is expressed from a broad spectrum of tissues and related to a plethora of physiological and pathophysiological processes, such as Alzheimer's disease, atherosclerosis and cancer. In its dominant form it is expressed as a secretory protein (secreted CLU, sCLU). During its maturation, the sCLU-precursor is N-glycosylated and cleaved into an α- and a β-chain, which are connected by five symmetrical disulfide bonds. Recently, it has been demonstrated that besides the predominant sCLU, rare intracellular CLU forms are expressed in stressed cells. Since these forms do not enter or complete the secretory pathway, they are not proteolytically modified and show either no or only core glycosylation. Due to their sparsity, these intracellular forms are functionally poorly characterized. To evaluate the function(s) of these stress-related intracellular forms, we investigate for the first time the impact of proteolytic cleavage, differential glycosylation and the influence of the redox environment on the chaperone activity of CLU. Methods: Non-cleavable sCLU was generated by expression from a mutant construct of sCLU, in which the furin-like proprotein convertase (PC) recognition site was modified. After purification of recombinant uncleaved sCLU from the medium of over-expressing cells, we performed chaperone activity assays to compare the activities of wild-type (cleaved) and uncleaved mutant sCLU. Additionally, this approach enabled us to investigate the role of carbohydrates, the proteolytic maturation and reducing conditions on CLU chaperone activity. Further, we characterized the differentially treated CLU forms by using MALDI-TOF, CD-spectroscopy and Western blotting in addition to the functional assay. Results: We show that the PC-cleavage is dispensable for sCLU chaperone activity. Moreover, our data demonstrate that while fully deglycosylated sCLU lacks chaperone activity, partially deglycosylated sCLU is still capable of solubilizing target proteins. Most importantly, we here demonstrate for the first time that uncleaved sCLU is highly sensitive towards reducing conditions. Conclusions: Our study provides evidence that unglycosylated intracellular CLU forms cannot exhibit a chaperone activity compared to sCLU. Additionally, we support recent postulates that glycosylated intracellular CLU forms may act as a redox sensor under oxidative stress conditions. Furthermore, we conclude that the proteolytic cleavage of sCLU is important to maintain full chaperone activity, i.e. in the presence of necrosis.

A new colubrid species of the genus Tantilla from the dry forest of the northern Peruvian Andes is described on the basis of two specimens, which exhibit a conspicuous sexual dimorphism. Tantilla tjiasmantoi sp. nov. represents the third species of the genus in Peru. The new species is easily distinguished from its congeners by the combination of scalation characteristics and the unusual transversely-banded color pattern on the dorsum. A detailed description of the skull morphology of the new species is given based on micro-computed tomography images. The habitat of this new species is gravely threatened due to human interventions. Conservation efforts are urgently needed in the inter-Andean valley of the Maranon River.

This work reports on the collection of the gekkonid species Phyllodactylus thompsoni and Phyllodactylus delsolari from seven and four new localities, respectively, extending the distribution of both species southward along the Río Marañón valley in Peru by about 130 km and the altitudinal range to nearly 1,900 m above sea level.

We herein describe a new colorful species of Polychrus with a conspicuous sexual dimorphism from the dry forest of the northern portion of Región de La Libertad, Peru. The new species differs from all other Polychrus species, in that this species has very small dorsal scales and thus a higher number of scales around midbody and in the middorsal line from behind the occipital scales to the level of the posterior edge of the thigh. Furthermore, we redescribe P. peruvianus whose original description is short and lacks information on intraspecific variation and sexual dimorphism. Also, we add some information on intraspecific variation and ecology of P. gutturosus. Finally, we synonymize P. spurrelli Boulenger with P. gutturosus.

Descrevemos aqui a reprodução de quatro lagartixas da familia Phyllodactylidae, Phyllopezus maranjonensis, Phyllodactylus delsolari, P. thompsoni e P. reissii, do Vale do Alto Marañon, Peru. As observações foram feitas entre março e maio de 2010, e os espécimes-testemunhos, depositados no Zoologisches Forschungsmuseum Alexander Koenig (ZFMK), foram radiografados para verificar a presença de fêmeas ovígeras. As fêmeas ovígeras de P. maranjonensis, P. delsolari e P. reissii portavam dois ovos, enquanto P. thompsoni produz um único ovo, que é depositado na serapilheira ou em frestas de paredões rochosos. As fêmeas de todas as espécies são mais provavelmente capazes de produzir desovas múltiplas ao longo do ano; o período reprodutivo parece prolongado em pelo menos duas espécies. Adicionalmente, medidas de diversos espécimes das quatro espécies são apresentadas aqui, revelando que Phyllopezus maranjonensis é a maior espécie de lagartixa do Novo Mundo.

A new concept for the development of semiconductor field-effect biosensors by modification of a gate surface with tobacco mosaic virus (TMV) nanotubes, serving as enzyme nanocarrier, is presented. TMV nanotubes enable an immobilization of a high amount of enzymes without substantial loss of their activity, resulting in an enhanced biosensor performance. This approach has been experimentally demonstrated by realizing a capacitive field-effect penicillin biosensor using TMV nanotubes functionalized with the enzyme penicillinase as model system.

Clusterin, also known as apolipoprotein J, is expressed from a variety of tissues and implicated in pathological disorders such as neurodegenerative diseases, ischemia and cancer. In contrast to secretory clusterin (sCLU), which acts as an extracellular chaperone, the synthesis, subcellular localization and function(s) of intracellular CLU isoforms is currently a matter of intense discussion. By investigating human CLU mRNAs we here unravel mechanisms leading to the synthesis of distinct CLU protein isoforms and analyze their subcellular localization and their impact on apoptosis and on NF-κB-activity. Quantitative PCR-analyses revealed the expression of four different stress-inducible CLU mRNA variants in non-cancer and cancer cell lines. In all cell lines variant 1 represents the most abundant mRNA, whereas all other variants collectively account for no more than 0.34% of total CLU mRNA, even under stressed conditions. Overexpression of CLU cDNAs combined with in vitro mutagenesis revealed distinct translational start sites including a so far uncharacterized non-canonical CUG start codon. We show that all exon 2-containing mRNAs encode sCLU and at least three non-glycosylated intracellular isoforms, CLU1‑449, CLU21‑449 and CLU34‑449, which all reside in the cytosol of unstressed and stressed HEK‑293 cells. The latter is the only form expressed from an alternatively spliced mRNA variant lacking exon 2. Functional analysis revealed that none of these cytosolic CLU forms modulate caspase-mediated intrinsic apoptosis or significantly affects TNF-α-induced NF-κB-activity. Therefore our data challenge some of the current ideas regarding the physiological functions of CLU isoforms in pathologies.

ABSTRACT Objectives: to characterize theses and dissertations that address Kangaroo-Mother Care Method produced in Brazilian Graduate Nursing Programs from 2000 to 2017. Methods: a documentary study of qualitative approach. From a total of 207 papers, 185 were excluded and 22 were selected. Results: in 2006 and 2014 there was a higher number of master’s dissertations. In 2001, 2013 and 2017, there were three productions of doctoral theses. Universidade Federal de Santa Catarina (Federal University of Santa Catarina) presented the largest number of publications, with five studies, followed by Universidade Federal da Bahia (Federal University of Bahia), with four. Final considerations: Kangaroo-Mother Care Method has been researched in Brazilian Graduate Nursing Programs, mainly from the qualitative approach, highlighting the importance, contributions and difficulties of its application as a public policy.