Paraparesis due to angio-neurotropic Gurltia paralysans in a domestic cat (Felis catus) and retrospective study on feline gurltiosis cases in South America.

Introduction: The nematode Gurltia paralysans is a neglected angio-neurotropic parasite causing chronic meningomyelitis in domestic cats (Felis catus) as well as wild felids of the genus Leopardus in South America. Adult G. paralysans nematodes parasitize the leptomeningeal veins of the subarachnoid space and/or meningeal veins of the spinal cord parenchyma. The geographic range of G. paralysans encompasses rural and peri-urban regions of Chile, Argentina, Uruguay, Colombia and Brazil. Methods: This case report presents clinical and pathological findings of a G. paralysans-infected cat suffering from severe thrombophlebitis and meningomyelitis resulting in ambulatory paraparesis. Neurological examination of affected cat localized the lesions at the thoracolumbar (T3-L3) and lumbosacral (L4-Cd4) segments. Molecular and morphological characteristics of extracted nematodes from parasitized spinal cord veins confirmed G. paralysans. Additionally, data obtained from a questionnaire answered by cat owners of 12 past feline gurltiosis cases (2014-2015) were here analyzed. Questionnaire collected data on age, gender, geographic location, type of food, hunting behavior, type of prey, and other epidemiological features of G. paralysans-infected cats. Results and Discussion: Data revealed that the majority of cats originated from rural settlements thereby showing outdoor life styles with hunting/predatory behaviors, being in close contact to wild life [i.e. gastropods, amphibians, reptiles, rodents, birds, and wild felids (Leopardus guinia)] and with minimal veterinary assistance. Overall, this neglected angio-neurotropic G. paralysans nematode still represents an important etiology of severe thrombophlebitis and meningomyelitis of domestic cats living in endemic rural areas with high biodiversity of definitive hosts (DH), intermediary (IH), and paratenic hosts (PH). The intention of this study is to generate awareness among veterinary surgeons as well as biologists on this neglected feline neuroparasitosis not only affecting domestic cats but also endangered wild felid species of the genus Leopardus within the South American continent.

Non-invasive detection of Orthohalarachne attenuata (Banks, 1910) and Orthohalarachne diminuata (Doetschman, 1944) (Acari: Halarachnidae) in free-ranging synanthropic South American sea lions Otaria flavescens (Shaw, 1800).

Respiratory mites of the genera Orthohalarachne and Halarachne (Acari: Halarachnidae) are causative agents of nasopharyngeal/nasopulmonary acariasis in pinnipeds and sea otters. Until now, these endoparasitic mites were mainly diagnosed via necropsies and invasive procedures. So far, non-invasive diagnostic techniques have neither been developed nor applied in free-ranging pinnipeds. In the current study, we aimed to evaluate the halarachnid mite infestation status of free-ranging "urban" South American sea lions Otaria flavescens in the city of Valdivia, Chile. Therefore, non-invasive sampling methods were used in the current study, e. g. by observation-based sampling of freshly expectorated nasal mucus in the animal environment. Further, collection devices were developed for target-oriented sampling of sneezed nasal mucus, including sterile petri dishes and stretched clingfilms mounted on telescopic rods. Applying these techniques, 26 individual sputum samples were collected. 11.5% of sputum samples proved positive for halarachnid larvae (in total, n = 7), which were morphologically identified as Orthohalarachne attenuata (n = 2) or Orthohalarachne diminuata (n = 5). In one of the individual sea lion mucus samples, both Orthohalarachne species were detected, thereby confirming a patent co-infestation in vivo. 16S rDNA-based molecular identification of individual Orthohalarachne spp. larvae confirmed morphological findings. For the first time, we here molecularly characterized Orthohalarachne spp. on the basis of three gene regions [18S, 28S and the internal transcribed spacer 1 (ITS1)]. Overall, current data include the successful application of non-invasive techniques to sample halarachnid mites from free-ranging synanthropic pinnipeds and contribute to the current knowledge on respiratory mites infesting South American sea lions by combining morphological and molecular methods to overcome challenges in species identification. This study should further serve as baseline study and calls for more research on occurrence, biology and health implications of orthohalarachnosis in free-living as well as captive pinnipeds.

Ku-DNA binding inhibitors modulate the DNA damage response in response to DNA double-strand breaks.

The DNA-dependent protein kinase (DNA-PK) plays a critical role in the DNA damage response (DDR) and non-homologous end joining (NHEJ) double-strand break (DSB) repair pathways. Consequently, DNA-PK is a validated therapeutic target for cancer treatment in certain DNA repair-deficient cancers and in combination with ionizing radiation (IR). We have previously reported the discovery and development of a novel class of DNA-PK inhibitors with a unique mechanism of action, blocking the Ku 70/80 heterodimer interaction with DNA. These Ku-DNA binding inhibitors (Ku-DBi's) display nanomolar activity in vitro, inhibit cellular DNA-PK, NHEJ-catalyzed DSB repair and sensitize non-small cell lung cancer (NSCLC) cells to DSB-inducing agents. In this study, we demonstrate that chemical inhibition of the Ku-DNA interaction potentiates the cellular effects of bleomycin and IR via p53 phosphorylation through the activation of the ATM pathway. This response is concomitant with a reduction of DNA-PK catalytic subunit (DNA-PKcs) autophosphorylation at S2056 and a time-dependent increase in H2AX phosphorylation at S139. These results are consistent with Ku-DBi's abrogating DNA-PKcs autophosphorylation to impact DSB repair and DDR signaling through a novel mechanism of action, and thus represent a promising anticancer therapeutic strategy in combination with DNA DSB-inducing agents.

Gurltia paralysans: A Neglected Angio-Neurotropic Parasite of Domestic Cats (Felis catus) and Free-Ranging Wild Felids (Leopardus spp.) in South America.

Gurltia paralysans is a neglected and re-emerging metastrongyloid angio-neurotropic nematode causing severe chronic meningomyelitis in domestic cats (Felis catus) as well as in free-ranging small wild felids such as kodkods (Leopardus guigna), margays (Leopardus wiedii) and the northern tiger cat (Leopardus triginus) in South America. Within these definitive hosts (DH), adult males and females of G. paralysans parasitize the leptomeningeal veins of the subarachnoid space and/or the meningeal veins of spinal cord parenchyma, inducing vascular alterations. Feline gurltiosis has been associated with progressive thrombophlebitis of the meningeal veins, resulting in ambulatory paraparesis, paraplegia, ataxia, hindlimb proprioceptive deficit, uni- or bilateral hyperactive patellar reflexes, faecal and urinary incontinence, and tail paralysis. The complete life cycle of G. paralysans has not been elucidated yet, but most probably involves gastropods as obligate intermediate hosts (IH). In terms of epidemiology, G. paralysans infections in domestic and wild felids are scattered around various South American countries, with hyperendemic areas in southern parts of Chile. Etiological diagnosis of G. paralysans still represents a challenge for clinicians due to a lack of evidence of the excretion of either eggs or larvae in faeces or in other body fluids. Diagnosis is based on clinical neurological signs, imaging findings through computed tomography (CT), myelography, magnetic resonance imaging (MRI), and post mortem examination. Nonetheless, novel diagnostic tools have been developed, including semi-nested PCR for detecting circulating G. paralysans DNA in the cerebrospinal fluid, serum and blood samples as well as in serological diagnostic kits detecting parasite-derived antigens, but these need validation for routine usage. The hypothetical life cycle of G. paralysans is addressed in this article, including the exogenous stages (i.e., eggs, and first- (L1), second- (L2) and third-stage (L3) larvae) and obligate gastropod IH and/or paratenic hosts (PH), and we propose possible anatomical migration routes of infective L3 that reach the leptomeningeal veins in vivo. Finally, the pro-inflammatory endothelium- and leukocyte-derived innate immune reactions of the host against G. paralysans, which most likely result in thrombophlebitis and meningomyelitis, are briefly touched on.

Morphological, Behavioral, and Molecular Characterization of Avian Schistosomes (Digenea: Schistosomatidae) in the Native Snail Chilina dombeyana (Chilinidae) from Southern Chile.

Avian schistosomes are blood flukes parasitizing aquatic birds and snails, which are responsible for a zoonotic disease known as cercarial dermatitis, a hypersensitive reaction associated to the cutaneous penetration of furcocercariae. Despite its worldwide distribution, its knowledge is fragmentary in the Neotropics, with most of data coming from Argentina and Brazil. In Chile, there are only two mentions of these parasites from birds, and one human outbreak was associated to the genus "Trichobilharzia". However, the identity of such parasites is pending. The aim of this study was to identify the furcocercariae of avian schistosomes from Southern Chile using an integrative approach. Thus, a total of 2283 freshwater snails from different families were collected from three different regions. All snails were stimulated for the shedding of furcocercariae, but only Chilina dombeyana (Chilinidae) from the Biobío region was found to be parasitized. The morphology and phylogenetic analyses of 28S and COI genes stated two lineages, different from Trichobilharzia, shared with Argentina. This study provides new information on Neotropical schistosomes, highlighting the need for major research on these neglected trematodes, which are considered to be emerging/re-emerging parasites in other parts of the globe as consequence of anthropogenic disturbances and climatic change. Highlights: 1. Two different lineages (Lineage I and II) were described and molecularly characterized (28S and COI genes); 2. Cercaria chilinae I y II are proposed as a synonymous of Lineage II. Thus, a total of four different lineages of avian schistosomes are related to Chilina spp.; 3. Chilina spp. represents an important intermediate host for avian schistosomes in South America, constituting a reservoir de schistosomes with zoonotic potential; 4. Coinfection between the two different lineages was found, a finding previously not reported for avian schistosomes; 5. Expansion in the geographic distribution of Nasusbilharzia melancorhypha from its original record in Argentina, with Chilina dombeyana as an additional intermediate host.

Revisión anatómica de la cavidad oral, glándulas salivares y esófago de dos ejemplares de pudú (Pudu puda) / Anatomical review of the oral cavity, salivary glands and esophagus of two specimens of pudú (Pudu puda)

RESUMEN: El pudú (Pudu puda), pertenece al orden Artiodactyla y a la familia Cervidae, corresponde al segundo cérvido más pequeño del mundo y es endémico de Chile y Argentina. Son herbívoros que se alimentan de hojas jóvenes y brotes de algunos árboles, arbustos, hierbas, frutos y flores. Esta especie está catalogada en estado Vulnerable, por lo que se prohíbe su caza y captura, esto debido a que su población ha disminuido sustancialmente a causa de la pérdida y fragmentación de la vegetación nativa, los incendios forestales, ataques de perros asilvestrados y caza furtiva. Existe escasa información científica anatómica de esta especie, y más aún referente a su morfología digestiva, por lo que se requieren estudios que aporten información específica de este cérvido. Se utilizaron dos cadáveres de pudú hembras adultas, los cuales fueron donados por el Servicio Agrícola y Ganadero (SAG) de Talca, Región del Maule, Chile. Dentro de los resultados, podemos indicar que la cavidad oral, glándulas salivares y esófago se asemejan a otros rumiantes domésticos, como también existen semejanzas con el venado de campo, sin embargo, existen diferencias como la forma y desarrollo de la glándula parótida; el conducto parotídeo pasa ventral al músculo masetero y se abre en una papila a nivel del tercer premolar superior; la glándula mandibular se divide en tres lobos; la glándula sublingual solo posee porción polistomática, las glándulas bucales dorsales poseen un menor tamaño que las bucales ventrales, la glándula bucal ventral es alargada y de forma compacta y se observa la presencia de una redondeada glándula bucal intermedia, poseen 11 crestas palatinas por lado en el paladar duro y además la papila incisiva se encuentra rodeando al pulvino dental. Esta información es relevante y corresponde a un aporte al conocimiento específico de esta especie, la cual es endémica, vulnerable y protegida.

SUMMARY: The pudu (Pudu puda), belongs to the Artiodactyla order and the Cervidae family, corresponding to the second smallest deer in the world and is endemic to Chile and Argentina. They are herbivores that feed on young leaves and shoots of certain trees, shrubs, herbs, fruits, and flowers. The status of is species is classified as Vulnerable; therefore, its hunting and capture is prohibited, this because its population has decreased substantially due to the loss and fragmentation of native vegetation, forest fires, attacks by feral dogs and poaching. There is little scientific anatomical information available on this species, and even more so regarding its digestive morphology, which is why studies are required to provide specific information on this deer. Two adult female pudu specimens were used, which were donated by the Agricultural and Livestock Service (SAG) of Talca, Maule Region, Chile. Among the results, we can indicate that the oral cavity, salivary glands and esophagus resemble other domestic ruminants; there are also similarities with the field deer, however, there are differences such as the shape and development of the parotid gland; the parotid duct passes ventral to the masseter muscle and opens into a papilla at the level of the upper third premolar; the mandibular gland is divided into three wolves; the sublingual gland only has a polystomatic portion, the dorsal buccal glands are smaller than the ventral buccal glands, the ventral buccal gland is elongated and compact and the presence of a rounded intermediate buccal gland is observed, it has 11 palatal ridges per side in the hard palate and also the incisive papilla is found surrounding the dental pulvino. This information is relevant and corresponds to a report on the specific knowledge of this species, which is endemic, vulnerable, and protected.

Morphological description of extrinsic muscles of the thoracic limb in a specimen of pudu (Pudu puda) / Descripción morfológica de la musculatura extrínseca del miembro torácico de un espécimen de pudú (Pudu puda)

SUMMARY: The pudu (Pudu puda) is classified as an artiodactyl of the Cervidae family. It is a native species found in Argentina and Chile. It is estimated that its population has been substantially reduced due to several causes, such as loss of forests, predation, hunting, and vehicle accident. Therefore, this species is protected due to its vulnerable conservation status. The extrinsic muscles of the thoracic limb have great importance in the biomechanical functionality of the suspension of the limb, neck, and head, as well as participating in the movement of the thoracic wall and thoracic limb. The objective of the present study is to describe the extrinsic musculature of the thoracic limb of a specimen of pudu, comparing the results with those described for domestic ruminants in the classical anatomical bibliography. Basic procedures: All extrinsic muscles were analyzed, describing shape, distribution, origin, and insertion. The results indicate that the pudu specimen has anatomical characteristics similar to domestic ruminants; however, some differences should be considered. Main findings: The brachiocephalicus muscle has an evident clavicular intersection that separates it into cleidobrachialis, cervicalis, and mastoideus pars. The pectoralis superficialis muscle has two distinct and independent pars, and the pectoralis profundus muscle has a partial fusion with the latissimus dorsi and cutaneous trunci muscles. The subclavius muscle is small and has an elongated shape and goes deep into the cephalic vein, just at the point it is a tributary of the external jugular vein. The results of this study present specific anatomical features of Pudu puda providing novel reference information and expanding scientific knowledge of this scarcely studied wild species.

RESUMEN: El pudu (Pudu puda) se clasifica como un artiodáctilo de la familia Cervidae. Es una especie nativa que se encuentra en Argentina y Chile y se estima que su población se ha reducido sustancialmente debido a varias causas, tal como la pérdida de bosques, depredación, caza y accidentes de vehículo. Debido a lo anterior esta especie está protegida por su estado de conservación vulnerable. Los músculos extrínsecos del miembro torácico tienen gran importancia en la funcionalidad biomecánica de la suspensión del miembro, el cuello y la cabeza, además de participar en el movimiento de la pared torácica y el miembro torácico. El objetivo del presente estudio es describir la musculatura extrínseca del miembro torácico de un ejemplar de pudu, comparando los resultados con los descritos para rumiantes domésticos en la bibliografía anatómica clásica. Se analizaron todos los músculos extrínsecos, describiendo la forma, distribución, origen e inserción. Los resultados indican que el espécimen de pudu tiene características anatómicas similares a las de los rumiantes domésticos; sin embargo, deben tenerse en cuenta algunas diferencias. Hallazgos principales: El músculo braquiocefálico tiene una interección clavicular evidente que lo separa en pars cleidobraquial, cervical y mastoideo. El músculo pectoral superficial tiene dos porciones distintas e independientes, y el músculo pectoral profundo tiene una fusión parcial con los músculos latissimus dorsi y cutáneo del tronco. El músculo subclavio es pequeño y tiene una forma alargada y se ubica profundamente a la vena cefálica, justo en el punto en el que es un afluente de la vena yugular externa. Los resultados de este estudio presentan características anatómicas específicas de Pudu puda proporcionando información de referencia novedosa y ampliando el conocimiento científico de esta especie silvestre escasamente estudiada.

Evaluation of terrestrial gastropods as possible intermediate hosts of Gurltia paralysans in southern Chile.

Gurltia paralysans is the causal agent of gurltiosis in domestic cats in South America. Although the life cycle of G. paralysans is unknown, it is thought that gastropods could act as intermediate hosts (IHs), as is the case for several nematodes in the Angiostrongylidae family. The aim of this study was to search for G. paralysans larvae in terrestrial gastropods and determine their role in the life cycle of this nematode species. Terrestrial gastropod samples (n=835) were collected in Punucapa, Valdivia, southern Chile, where cases of gurltiosis had been reported before. The samples included species from the families Arionidae, Limacidae, Helicidae and Milacidae. All gastropods were subjected to enzymatic digestion to isolate G. paralysans larvae. Ten percent of the gastropod samples were analyzed using seminested PCR targeting the 28S rRNA gene, while 2.6% were analyzed by histopathological examination. The results indicated the absence of G. paralysans when using any of the three methods. In conclusion, further studies are needed to evaluate specific species of aquatic or native gastropods acting as possible IHs (in this geographic location).

Expansion in the latitudinal distribution of Rhipicephalus sanguineus sensu stricto (Acari: Ixodidae) to southern Chile.

Rhipicephalus sanguineus is a species complex with a challenging diagnosis. There are two main lineages: a temperate lineage-now recognized as R. sanguineus sensu stricto (s.s.)-and a tropical lineage. In the Americas, several studies examined the distribution of both lineages. Meanwhile, in Chile, the distribution of the tropical lineage was reported from 18° to 22° S and R. sanguineus s.s. from 20° to 33° S; the morphological diagnosis located R. sanguineus in south-central Chile. In this study, we assessed the morphological and genetic identity of ticks collected from 47 dogs in southern Chile. Six dogs (12.8%) were parasitized by ticks that were morphologically and genetically assigned to R. sanguineus s.s. This finding would represent the southernmost record of this species in the country, located at over 308 km further south than the previous record, suggesting a latitudinal expansion of its distribution. Experimental studies are further required to understand how this tick survives extreme weather conditions when compared to other R. sanguineus s.s. from lesser latitudes.

Molecular survey of Bartonella spp. and haemoplasmas in American minks (Neovison vison).

The aim of this study was to perform a molecular survey and characterize Bartonella spp. and haemotropic Mycoplasma (haemoplasmas) in invasive American minks (Neovison vison) from Southern Chile. Additionally, we addressed risk factors for positivity in both groups of agents. Blood and/or tissue samples from 246 minks were analysed by qPCR targeting the nuoG gene for Bartonella spp. and conventional (c)PCR for 16S rRNA for haemotropic Mycoplasma spp. nuoG qPCR-positive Bartonella spp. samples were submitted to cPCR assays (ITS, ribC, gltA, rpoB, pap-31 and ftsZ genes) to perform phylogenetic inferences. Haemotropic Mycoplasma spp. 16S-positive samples were further amplified by cPCR targeting RNaseP gene (160-210 bp) and by two overlapping 16S rRNA cPCR assays to amplify a larger portion of the gene (1,200bp) for phylogenetics. Bartonella DNA was detected in 8.9% of minks (22/246). Out of 22 nuoG qPCR-positive samples, one and two showed positive results in cPCR assays based on ITS and ribC, respectively. Consistent sequencing results were obtained for only one ITS sample (464 bp sequence), which shared 99.6% identity with B. clarridgeiae. Two per cent of minks (5/246) were positive for 16S rRNA haemotropic Mycoplasma-cPCR assay. Two concatenated sequences of 16S rRNA (1,176 and 1,230 bp) were obtained: one sample shared 97.87% identity with haemotropic Mycoplasma sp. from a wild rodent, and the other 96.49% identity with 'Candidatus Mycoplasma haematoparvum' from a dog. All BLAST results were supported by phylogenetic analysis. One haemoplasma RNase P sequence shared 94.86% identity with Mycoplasma haemofelis from a cat. No risk factors for PCR positivity were identified. In a nutshell, Bartonella clarridgeiae and a potentially novel haemoplasma closely related to haemoplasmas previously reported in rodents, dogs, domestic and wild cats were described for the first time in American minks.

Evaluation of terrestrial gastropods as possible intermediate hosts of Gurltia paralysans in southern Chile / Avaliação de gastrópodes terrestres como possíveis hospedeiros intermediários de Gurltia paralysans no sul do Chile

Abstract Gurltia paralysans is the causal agent of gurltiosis in domestic cats in South America. Although the life cycle of G. paralysans is unknown, it is thought that gastropods could act as intermediate hosts (IHs), as is the case for several nematodes in the Angiostrongylidae family. The aim of this study was to search for G. paralysans larvae in terrestrial gastropods and determine their role in the life cycle of this nematode species. Terrestrial gastropod samples (n=835) were collected in Punucapa, Valdivia, southern Chile, where cases of gurltiosis had been reported before. The samples included species from the families Arionidae, Limacidae, Helicidae and Milacidae. All gastropods were subjected to enzymatic digestion to isolate G. paralysans larvae. Ten percent of the gastropod samples were analyzed using seminested PCR targeting the 28S rRNA gene, while 2.6% were analyzed by histopathological examination. The results indicated the absence of G. paralysans when using any of the three methods. In conclusion, further studies are needed to evaluate specific species of aquatic or native gastropods acting as possible IHs (in this geographic location).

Resumo Gurltia paralysans é o agente etiológico da gurltiose em gatos domésticos na América do Sul. Embora o ciclo biologico de G. paralysans seja desconhecido, provavelmente é indireto com gastrópodes atuando como hospedeiros intermediários (HIs), como no caso de vários nematoides da família Angiostrongylidae. O objetivo deste estudo foi investigar a presença de larvas de G. paralysans em gastrópodes terrestres para avaliar seu papel no ciclo de vida do parasito. Amostras de gastrópodes terrestres (n = 835) foram coletadas em Punucapa, Valdivia, sul do Chile, onde casos de gurltiose foram relatados anteriormente. As amostras incluíram espécies das famílias Arionidae, Limacidae, Helicidae e Milacidae. Todos os gastrópodes foram submetidos à digestão enzimática para isolar as larvas de G. paralysans. 10% das amostras foram analisadas, utilizando-se seminested PCR para o gen 28S RNAr de G. paralysans, enquanto 2,6% foram analisados ​​por exame histopatológico. Os resultados indicaram ausência de G. paralysans em todos os três métodos. Os dados permitem concluir que são necessários mais estudos para avaliar espécies específicas de gastrópodes aquáticos ou nativos, que atuam como possíveis hospedeiros intermediários nessa localização geográfica.

Intra Vitam Diagnosis of Neglected Gurltia paralysans Infections in Domestic Cats (Felis catus) by a Commercial Serology Test for Canine Angiostrongylosis and Insights into Clinical and Histopathological Findings-Four-Case Report.

Gurltia paralysans is a metastrongyloid nematode which belongs to the Angiostrongylidae family and presents tropism for veins of the subarachnoid space in vivo of domestic and wild felids causing a progressive and chronic clinical manifestation of paraparesis/paraplegia. The geographic distribution of G. paralysans includes rural and periurban areas of South America and was recently reported in Europe. To date, a definitive diagnosis of feline gurltiosis is still conducted by post-mortem examination of the spinal cord in affected animals. A presumptive diagnosis of feline gurltiosis can also be achieved based on data of compatible clinical signs and associated epidemiological risk factors. The aim of this preliminary study was to evaluate the commercial serological test Angio Detect TM® (IDEXX Laboratories) as a possible diagnostic method of feline gurltiosis in vivo. For the study, 10 domestic felines (Felis catus) which originated from a high endemic area of Southern Chile, were analyzed. All felines presented chronic paraparesis or severe paraplegia. Subsequently, commercial Angio Detect TM® serological tests for the detection of closely related Angiostrongylus vasorum in canids were performed according to manufacturer's instructions. Conducted serological tests were positive in seven of ten felines showing paraplegia/paraparesis and presumably infected with G. paralysans, and four of them were additionally necropsied, and presented macroscopic findings compatible with feline gurltiosis. Furthermore, the presence of adult female and male G. paralysans specimens at the level of the subarachnoid vasculature in affected spinal cord segments were observed during necropsy. Histopathology demonstrated severe eosinophilic meningomyelitis, coagulopathies with thrombosis in G. paralysans-parasitized leptomeningeal veins. Results of this preliminary study suggest a cross-reaction between A. vasorum-specific antigens, which also parasitize blood vessels in vivo, and G. paralysans when using an Angio Detect TM® test, which suggests its helpful use as a new diagnostic method for feline gurltiosis in live domestic felines. Additional specific antigen research will be required in order to better understand the sensitivity and specificity of A. vasorum antigens used in this test and for existing cross-reactivity with G. paralysans-derived antigens for future a suitable intra vitam immunodiagnosis of neglected feline gurltiosis.

Discovery and development of novel DNA-PK inhibitors by targeting the unique Ku-DNA interaction.

DNA-dependent protein kinase (DNA-PK) plays a critical role in the non-homologous end joining (NHEJ) repair pathway and the DNA damage response (DDR). DNA-PK has therefore been pursued for the development of anti-cancer therapeutics in combination with ionizing radiation (IR). We report the discovery of a new class of DNA-PK inhibitors that act via a novel mechanism of action, inhibition of the Ku-DNA interaction. We have developed a series of highly potent and specific Ku-DNA binding inhibitors (Ku-DBi's) that block the Ku-DNA interaction and inhibit DNA-PK kinase activity. Ku-DBi's directly interact with the Ku and inhibit in vitro NHEJ, cellular NHEJ, and potentiate the cellular activity of radiomimetic agents and IR. Analysis of Ku-null cells demonstrates that Ku-DBi's cellular activity is a direct result of Ku inhibition, as Ku-null cells are insensitive to Ku-DBi's. The utility of Ku-DBi's was also revealed in a CRISPR gene-editing model where we demonstrate that the efficiency of gene insertion events was increased in cells pre-treated with Ku-DBi's, consistent with inhibition of NHEJ and activation of homologous recombination to facilitate gene insertion. These data demonstrate the discovery and application of new series of compounds that modulate DNA repair pathways via a unique mechanism of action.

Physiological and metabolic responses to hypersalinity reveal interpopulation tolerance in the green macroalga Ulva compressa with different pollution histories.

There is scarce investigation addressing interpopulation tolerance responses to address the influence of a history of chronic stress exposure, as that occurring in polluted environments, in photoautotrophs. We evaluated ecophysiological (photosynthetic activity) and metabolic (oxidative stress and damage) responses of two populations of green macroalga Ulva compressa from polluted (Ventanas) and non-polluted (Cachagua) localions of central Chile, and exposed to controlled hypersalinity conditions of 32 (control), 42, 62 and 82 psu (practical salinity units) for 6 h, 48 h and 6 d. Both primary production (ETRmax) and photosynthetic efficiency (αETR) were generally higher in the population from Cachagua compared to Ventanas at all times and salinities. Moreover, at most experimental times and salinities the population from Ventanas had greater levels of H2O2 and lipid peroxidation that individuals from Cachagua. Total ascorbate was higher in the population of Cachagua than Ventanas at 42 and 82 psu after 6 and 48 h, respectively, while at 6 d concentrations were similar between both populations at all salinities. Total glutathione was greater in both populations after 6 h at all salinities, but at 48 h its concentrations were higher only in the population from Cachagua, a trend that was maintained at 6 d under 82 psu only. Reduced and oxidized ascorbate (ASC and DHA, respectively) and glutathione (GSH and GSSG, respectively) demonstrated similar patterns between U. compressa populations, with an increase oxidation with greater salinities but efficient recycling to maintain sufficient batch of ASC and GSH. When assessing the expression of antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and dehydroascorbate reductase (DHAR), while the population of Ventanas displayed a general trend of upregulation with increasing salinities along the experiments, U. compressa from Cachagua revealed patterns of downregulation. Results demonstrated that although both populations were still viable after the applied hypersalinities during all experimental times, biological performance was usually more affected in the population from the Ventanas than Cachagua, likely due to a depressed baseline metabolism after a long history of exposition to environmental pollution.

Molecular Detection of Gurltia paralysans by Semi-Nested PCR in Cerebrospinal Fluid and Serum Samples from Domestic Cats (Felis catus).

Gurltia paralysans is an angio-neurotropic metastrongyloid nematode that infects domestic and wild cats, invading the veins of the subarachnoid space of the spinal cord and mainly causing progressive paralysis of the pelvic limbs. The definitive diagnosis of feline gurltiosis can only be achieved by post-mortem examination that reveals the presence of the nematode in the spinal cord vein vasculature. An early diagnosis with conclusive results is required since laboratory and imaging findings are not sufficient. Therefore, the purpose of this study was to detect the presence of G. paralysans, via semi-nested PCR, in samples of cerebrospinal fluid (CSF) and the sera of domestic cats naturally infected with the parasite. A total of 12 cats with a diagnosis suggestive of feline gurltiosis were selected, and they underwent a complete neurological and imaging examination. DNA samples were analysed by semi-nested PCR, with universal (AaGp28Sa1/AaGp28Ss1) and specific (Gp28Sa3/Aa28Ss2) primers, for G. paralysans (G. paralysans 18S rRNA gene, partial sequence; ITS 1, 5.8S rRNA gene, and ITS 2, complete sequence; and 28S rRNA gene, partial sequence) and Aelurostrongylus abstrusus, obtaining amplifications of 356 and 300 bp, which indicated the presence or absence of nematode DNA, respectively. The presence of G. paralysans was detected in the CSF of four out of nine cats, and the sera of seven out of seven cats. In the sera analysis of five out of seven cats, a mixed infection with A. abstrusus was found, despite no alterations of the respiratory tract being observed during the necropsies. It is proposed that serum samples could be more effective than CSF in detecting the parasite by PCR analysis. Sequencing analysis showed high percentages of identity with G. paralysans, which indicated the feasibility of detection and the sensitivity/specificity of the method used, suggesting the implementation of semi-nested PCR as a routine diagnostic test for early and timely detection of feline gurltiosis.

Comparing quality of care in medical specialties between nurse practitioners and physicians.

BACKGROUND: The American health care system is facing a growing health care provider shortage in primary and specialty care settings. Research has established that nurse practitioners (NPs) match or exceed their physician colleagues in providing quality care in primary care settings. OBJECTIVE: This systematic review aimed to compare the quality of NP versus physician-led care in outpatient specialty care setting for clinical outcomes patient satisfaction. DATA SOURCES: The Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement guided the literature search (CINAHL, PubMed, and Cochrane Library) and interpretation of findings. Of the 228 articles that met the inclusion/exclusion criteria, 11 were selected for further review. CONCLUSIONS: Studies were conducted from 1995 to 2016 across four countries and spanned 10 distinct medical specialties. As a whole, these studies demonstrated that NPs in specialty settings perform as well as physicians terms of clinical safety and positive patient outcomes. Nurse practitioners matched or exceeded their physician counterparts in patient education and satisfaction. IMPLICATIONS FOR PRACTICE: Nurse practitioners are a feasible option for addressing specialty care shortages. Further research should investigate whether NPs and physicians are equally prepared to provide equivalent care immediately following their respective postbaccalaureate programs. If not, studies should explore specific training duration and elements NPs require to provide equivalent care.

Descripción morfológica y morfométrica de la miología intrínseca del miembro torácico en un ejemplar de pudú (Pudu puda) / Morphological and morphometric description of thoracic limb intrinsic myology in a specimen of pudu (Pudu puda)

El pudú (Pudu puda) se clasifica como un artiodáctilo de la familia Cervidae y es el ciervo más pequeño. Se estima que su población se ha reducido, esto debido a varias causas, como son la pérdida de los bosques, depredación, caza y atropellos. El objetivo del presente estudio es describir la musculatura intrínseca del miembro torácico de un ejemplar de pudú, comparando los resultados observados con lo descrito para rumiantes domésticos en la bibliografía anatómica clásica. Se estudió la musculatura desde proximal hacia distal, dividiendo al miembro en cuatro regiones topográficas: región escapular, braquial, antebraquial y de la mano. Además, se realizó un estudio morfométrico indicando el largo, ancho y grosor de los músculos. Los resultados indican que el ejemplar de pudú posee características anatómicas similares a rumiantes domésticos, sin embargo, se deben considerar algunas diferencias, tales como: la porción escapular del m. deltoides se une de manera aponeurótica con el m. infraespinoso; las dos cabezas del m. flexor ulnar del carpo se encuentran altamente fusionadas; no se observó el tercer tendón del m. bíceps braquial; el m. tríceps braquial es bien desarrollado y presenta cuatro cabezas musculares bien diferenciadas; el m. ancóneo permanece de manera independiente de la cabeza lateral del m. tríceps; el m. flexor digital profundo presenta cuatro cabezas y el m. interóseo es macroscópicamente tendinoso. La información entregada en este estudio corresponde a un aporte anatómico que permite ampliar el conocimiento científico de esta especie que se encuentra poco estudiada, protegida y vulnerable.

The pudú (Pudu puda) is classified as an artiodactyl and is the smallest deer of the Cervidae family. It is estimated that its population has been reduced for a number of reasons, such as loss of forests, predation and hunting. The objective of this study is to describe the intrinsic musculature of the thoracic limb of a Pudu specimen, comparing the results with those described for domestic ruminants in anatomical literature. Musculature was studied from proximal to distal, dividing the limb into four topographic regions: scapular, brachial, antebrachial and hand regions. In addition, a morphometric study was conducted indicating the length, width and thickness of the muscles. The results indicate that the Pudu species has anatomical characteristics similar to those of domestic ruminants. However, some differences should be considered, such as the scapular portion of M. deltoides joins aponeurotically with M. infraspinatus; the two heads of M. carpis ulnaris are highly fused; the third tendon of M. biceps brachii; the M. triceps brachii is well developed and has four distinct muscle heads; the M. Anconeus remains independent of the lateral head of M. triceps brachii; the M. digitorum profundus features four heads and the M. interosseous is macroscopically tendinous. The information provided in this study corresponds to an anatomical contribution that allows furthering scientific knowledge of this protected and vulnerable species, which is scarcely studied.

Gastrointestinal helminths and related histopathological lesions in black-necked swans Cygnus melancoryphus from the Carlos Anwandter Nature Sanctuary, Southern Chile / Helmintos gastrintestinais e lesões histopatológicas relacionadas em cisnes de pescoço negro Cygnus melancoryphus do Santuário da Natureza Carlos Anwandter, Sul do Chile

Abstract Black-necked swans are distributed across South America and face conservation problems in Chile according to data of the State institution SAG. The aim of this study was to identify helminths and to assess associated tissue damage via histopathology. A total of 19,291 parasites were isolated from 21 examined birds; 17 species were identified, including nematodes, flukes, and tapeworms. Of these, 12 were new host records, 13 were reported for the first time in Chile, and 5 were new records for the Neotropical region. Further, the flukes Schistosomatidae gen. sp. and Echinostoma echinatum are of zoonotic concern. Regarding histopathology, an inflammatory response was found along the birds' entire digestive tract. Nevertheless, it is difficult to declare that there is a clear association between such lesions and isolated parasites, as other noxa could be responsible as well. Although in some cases there was an evident association, such inflammatory responses and necrosis were minimal, as occurred with Capillaria, Retinometra, Catatropis, Echinostoma, and Schistosomatidae gen. sp. Nevertheless, Epomidiostomum vogelsangi caused granulomatous injuries, an important inflammatory response, and necrosis, but it always circumscribed to superficial layers of the gizzard. Conversely, Paramonostomum was not associated with an inflammatory response despite a high parasitic load.

Resumo O cisne de pescoço negro é distribuído por toda a América do Sul, e enfrenta problemas de conservação no Chile, sendo protegido pela Lei Estadual de Caça. O objetivo deste estudo foi identificar helmintos em cisnes e avaliar o dano tecidual por meio de histopatologia. Um total de 19.291 parasitas foi isolado de 21 aves examinadas, sendo 17 espécies identificadas, entre nematóides, trematódeos e tênias. Destes, 12 são novos registros de hospedeiros, 13 são reportados pela primeira vez no Chile, e 5 são novos registros para a região Neotropical. Além disso, os trematódeos Schistosomatidae gen. sp. e Echinostoma echinatum detectados têm importância zoonótica. Em relação à histopatologia, uma resposta inflamatória foi encontrada em todo o trato digestivo. Entretanto, é difícil estabelecer uma associação estrita de tais lesões com parasitas isolados, porque outros fatores também poderiam ser responsáveis. Em alguns casos, houve uma associação óbvia entre parasitas e lesões, embora a resposta inflamatória e a necrose fossem mínimas, como foi o caso dos gêneros Capillaria, Retinometra, Catatropis, Echinostoma e Schistosomatidae gen. sp. Entretanto, Epomidiostomum vogelsangi causou lesões granulomatosas com importante resposta inflamatória e necrose, mas sempre circunscrita às camadas superficiais da moela. Por outro lado, Paramonostomum não foi associado com uma resposta inflamatória óbvia apesar da alta carga parasitária.

Gastrointestinal helminths and related histopathological lesions in black-necked swans Cygnus melancoryphus from the Carlos Anwandter Nature Sanctuary, Southern Chile.

Black-necked swans are distributed across South America and face conservation problems in Chile according to data of the State institution SAG. The aim of this study was to identify helminths and to assess associated tissue damage via histopathology. A total of 19,291 parasites were isolated from 21 examined birds; 17 species were identified, including nematodes, flukes, and tapeworms. Of these, 12 were new host records, 13 were reported for the first time in Chile, and 5 were new records for the Neotropical region. Further, the flukes Schistosomatidae gen. sp. and Echinostoma echinatum are of zoonotic concern. Regarding histopathology, an inflammatory response was found along the birds' entire digestive tract. Nevertheless, it is difficult to declare that there is a clear association between such lesions and isolated parasites, as other noxa could be responsible as well. Although in some cases there was an evident association, such inflammatory responses and necrosis were minimal, as occurred with Capillaria, Retinometra, Catatropis, Echinostoma, and Schistosomatidae gen. sp. Nevertheless, Epomidiostomum vogelsangi caused granulomatous injuries, an important inflammatory response, and necrosis, but it always circumscribed to superficial layers of the gizzard. Conversely, Paramonostomum was not associated with an inflammatory response despite a high parasitic load.

MAPK Pathway under Chronic Copper Excess in Green Macroalgae (Chlorophyta): Involvement in the Regulation of Detoxification Mechanisms.

Following the physiological complementary/parallel Celis-Plá et al., by inhibiting extracellular signal regulated kinases (ERK), c-Jun N-terminal kinases (JNK), and cytokinin specific binding protein (p38), we assessed the role of the mitogen-activated protein kinases (MAPK) pathway in detoxification responses mediated by chronic copper (10 µM) in U. compressa. Parameters were taken at 6, 24, and 48 h, and 6 days (d). H2O2 and lipid peroxidation under copper and inhibition of ERK, JNK, or p38 alone increased but recovered by the sixth day. By blocking two or more MAPKs under copper, H2O2 and lipid peroxidation decayed even below controls. Inhibition of more than one MAPK (at 6 d) caused a decrease in total glutathione (reduced glutathione (GSH) + oxidised glutathione (GSSG)) and ascorbate (reduced ascorbate (ASC) + dehydroascorbate (DHA)), although in the latter it did not occur when the whole MAPK was blocked. Catalase (CAT), superoxide dismutase (SOD), thioredoxin (TRX) ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), and glutathione synthase (GS), were downregulated when blocking more than one MAPK pathway. When one MAPK pathway was blocked under copper, a recovery and even enhancement of detoxification mechanisms was observed, likely due to crosstalk within the MAPKs and/or other signalling processes. In contrast, when more than one MAPK pathway were blocked under copper, impairment of detoxification defences occurred, demonstrating that MAPKs were key signalling mechanisms for detoxification in macroalgae.