The pine shoot beetle Tomicus piniperda as a plausible vector of Fusarium circinatum in northern Spain

International audienceAbstractKey messageThe pine shoot beetle, Tomicus piniperda, is a potential vector of the pitch canker disease pathogen. The insect could transmit the pathogenic fungus during its maturation or regeneration feeding on the shoots of healthy pine crowns.ContextFusarium circinatum, the causal agent of pitch canker disease, currently affects Pinus radiata in northern Spain, causing pitch-soaked cankers and tree death. Although several species of the family Scolytinae have been reported as vectors of this pathogen, the role of the pine shoot beetle T. piniperda remains unclear.AimsThe general objective of this study was to determine whether T. piniperda is a vector for the pitch canker pathogen F. circinatum. For this purpose, Leach’s postulates (1) an association between T. piniperda and trees affected by pitch canker disease; (2) regular visits by T. piniperda to healthy P. radiata trees; (3) presence of the pathogen on the insect in nature; and (4) transmission of the pathogen to disease-free host material under controlled conditions.MethodsFresh green shoots with feeding galleries were collected from the ground, breeding galleries were collected from diseased trunks and insects were collected during their dispersion flights. A laboratory experiment was conducted in which specimens of T. piniperda were inoculated with the pathogen prior to feeding on shoots.ResultsIn the field, T. piniperda was found to be associated with both diseased and healthy P. radiata trees, and F. circinatum was found to be present, at low rates, on the exoskeleton of T. piniperda. In the laboratory experiment, evidence of the ability of T. piniperda to transfer the pathogen to healthy shoots was found.ConclusionsThe study findings indicate T. piniperda as a plausible vector of this pathogen. We postulate for the first time a potential relationship between the life cycles of T. piniperda and F. circinatum

Mapeo de Actores y Necesidades de Información Agroclimática en los Cultivos de Maíz y Frijol en sitios piloto - Colombia

Este trabajo presenta los resultados del mapeo de actores y necesidades de información agroclimática en los cultivos transitorios de seguridad alimentaria: maíz y frijol arbustivo en sitios piloto en Colombia. A través del mapeo de los actores que transmiten información a los agricultores, la confianza en la información proporcionada, y las necesidades de información para la toma de decisiones, logramos entender el ancho espectro de problemas y prioridades a tomar en cuenta en la producción de estos cultivos, y proponer soluciones útiles, relevantes, confiables, sostenibles y aplicables. Con este enfoque, se llevaron a cabo 27 entrevistas semi-estructuradas con informantes clave y 12 grupos focales con un total de 151 agricultores en los departamentos de Córdoba y Santander. Los resultados evidencian que una de las prioridades de los agricultores es recibir información agroclimática confiable a escala local, que permita tomar mejores decisiones de siembra, variedades, insumos y manejo de suelos y cultivos ante eventos de variabilidad climática. Se evidencia una desconfianza en las instituciones oficiales que transmiten información climática y un sentimiento difuso de falta de interés de las instituciones hacia los pequeños agricultores. Los agricultores se endeudan para lograr producir fríjol arbustivo y maíz o subsidian la producción por otros cultivos, mientras el acceso a un seguro agrícola es limitado. El formato preferido para recibir información climática es a través de mensajes de texto (celular) y mensajes en la radio local. Grupos de interacción presencial o virtual entre agricultores y expertos parecen una alternativa útil y confiable para recibir y compartir información en tiempo real. Sin embargo, los agricultores están interesados en adoptar el manejo agrícola tras un pronóstico si: a) la información es local, b) proviene de una fuente considerada confiable como expertos u otros agricultores, c) se transmite en el momento adecuado para la toma de decisiones y d) si sus efectos son medibles, verificables y validados, por ejemplo, a través de resultados de pilotos o parcelas experimentales en dos años consecutivos. ENGLISH This working paper presents the results of the mapping of actors and of agroclimatic information needs in pilot sites in Colombia for two annual crops, fundamental for food security: maize and bush beans. The mapping of the actors that transmit information to farmers, the confidence the latter have in the information provided, and their information needs for decision-making, allows us to understand the broad spectrum of issues and priorities to be taken into account in the production of these crops, and suggest solutions that are useful, relevant, reliable, sustainable and applicable. Under this assumption, we conducted 27 semi-structured interviews with key informants and 12 focus groups with a total of 151 farmers in the departments of Córdoba and Santander. Results show that one of the priorities of farmers is to receive reliable local agroclimatic information that enables them to take decisions on planting, varieties, inputs, soil and crop management, that address climate variability. We find a generalized distrust of official institutions that transmit weather information, and a feeling that institutions are not interested in smallholders. Farmers resort to indebtedness to produce beans and maize, or subsidize production with cash crops, while access to agricultural insurance is limited. The format preferred to receive weather information is text messages (phone) and information through the local radio. The creation of groups of farmers and experts who interact in person or virtually seem a useful and reliable alternative to receive and share information in real time. However, farmers are only interested in adopting farm management changes after a forecast if: a) the information is local, b) the information is given by a trusted source such as experts or other farmers, c) the information is transmitted at the right time for decision-making d) the effects of taking these decisions are measurable, verifiable and validated, for example, through pilots or results from experimental plots in two consecutive years

Local, reliable and timely agro-climatic information: a requirement of Colombian farmers

Awareness of climatic information relevant for future decisions through climatic forecasts is not enough to support farmer’s decision-making. It is necessary to look for agricultural management alternatives that allow farmers to adapt to climatic conditions and implement efficient and sustainable management systems, within a framework of effective communication with farmers (Jones, 2003; Pulwarty et al., 2003). In Latin America are very few the initiatives related to climatic forecasts. The Project “Agroclimatic services and food security information for improved decision-making – AgroClimas” aims to support private and public sector actors in Colombia, Guatemala and Honduras to implement and use agro-climatic risk management tools, validated and adjusted to the needs of final users. This info note presents results from a mapping of actors and agro-climatic information needs in crops important for food security (maize and bush beans), in pilot sites of the project in Colombia. The hypothesis underlying the study is that through mapping actors who transmit information (climatic, prices, technical and financial) to farmers, knowing if this information is considered reliable, identifying unattended information needs and preferred formats to receive it that would support production decisions, one can devise useful, relevant, reliable, sustainable applicable solutions

Differentiation and multipotential characteristics of mesenchymal stem cells derived from adipose tissue of an endangered wild cat (Leopardus guigna)

Adipose tissue derived mesenchymal stem cells (AMSCs) had been isolated and used for cell therapy in domestic cats. For wild cats, the isolation of AMSCs has only been reported in the black-footed cat (Felis nigripes). AMSCs obtained from wild cats may be useful to treat injuries of endangered cat species that remain in captivity or arrive at wildlife rehabilitation centers. Additionally, AMSCs might allow improvement of cloning techniques or assist in derivation of induced pluripotent stem cells. Endangered wild cats such as the guigna (Leopardus guigna), an endemic and endangered species from Chile and Argentina, might benefit greatly from the development of novel treatments or techniques that can be applied for its conservation. The objective of this study was to characterise putative AMSCs from guigna in terms of their main biological attributes, particularly, growth kinetics, differentiation ability and surface marker expression. Results obtained from this characterisation were compared with AMSCs isolated from domestic cats. AMSCs were isolated from peritoneal adipose tissue of female cats and subcutaneous tissue from a female guigna. Migration potential, colony-forming unit assay, mesodermal differentiation and surface marker expression (CD45, CD44, CD90, MHCI and MHCII) were evaluated. Domestic cat and guigna AMSCs displayed similar growth properties in culture. Both AMSC types showed mesodermal differentiation potential, in vitro homing potential and similar surface marker expression. These results indicate that AMSCs from subcutaneous tissue of guigna could have potential use as regenerative treatment for this species and might be considered for use in other biotechnological applications

Diagnóstico patológico y molecular de un caso de tuberculosis laríngea primaria en un médico

Primary laryngeal tuberculosis is a rare disease with a wide variety of clinical manifestations. We report the case of an ear-nose and throat diabetic physician with primary laryngeal tuberculosis. The diagnosis of laryngeal cancer had been considered before in another institution, and at the patient’s admission, he only presented dysphonia with a six-month evolution. The vocal cord histological examination showed granulomas, giant cells, fibrosis and necrosis, and the Ziehl-Neelsen staining showed acid-alcohol resistant bacilli. The culture from vocal cord tissue was positive for mycobacteria, and IS6110-PCR was positive too. The Mycobacterium tuberculosis strain was sensitive to first line drugs. Treatment using directly observed therapy short-course (DOTS) was initiated. Clinical presentation, differential diagnosis, diagnostic workup, associated risk factors and a brief literature revision are discussed in this article.La tuberculosis laríngea primaria es una enfermedad rara con una amplia gama de manifestaciones clínicas. Se reporta el caso de un médico otorrinolaringólogo con diabetes de tipo II y con diagnóstico de tuberculosis laríngea primaria. El diagnóstico de cáncer de cuerda vocal se había considerado en otra institución, y al ingreso el paciente sólo presentaba disfonía de seis meses de evolución. En el estudio histopatológico se encontraron granulomas, células gigantes multinucleadas, fibrosis y necrosis, y en la tinción de Ziehl-Neelsen, bacilos ácido-alcohol resistentes. El cultivo del tejido de cuerda vocal para micobacterias y la PCR-IS6110 fueron positivos. La cepa de Mycobacterium tuberculosis demostró sensibilidad a todos los medicamentos de primera línea. Se inició tratamiento según la estrategia DOTS (Directly Observed Treatment, Short-course). Se discute la presentación clínica, los diagnósticos diferenciales, el abordaje diagnóstico y los factores de riesgo asociados y se hace una breve revisión de la literatura.

Detekcija polimorfizama jednog nukleotida (SNP-a) u HER2, MUC1, ESR1 i BRCA1 gena povezanih s tumorom mliječne žlijezde kuja

Worldwide, canine mammary cancer (CMC) is the most frequent type of neoplasia in female dogs, and it is three times more frequent in dogs than in humans. In Colombia, CMC is the second most frequent type of cancer, after skin neoplasia. Genetics is one of the most important factors involved in any type of cancer, and the genetic basis of this disease is reflected through line breeding due to changes in allelic frequencies that are traceable using molecular markers. This study aimed to detect single nucleotide polymorphisms (SNPs) associated with CMC in blood samples collected from collected from healthy and CMC female dogs at Diego Villegas Toro Veterinary Hospital of Universidad de Caldas (Manizales, Colombia). We designed primers using Primer-BLAST and Primer3, and gene fragments from HER2, MUC1, ESR1, and BRCA1 were amplified to identify SNPs through genome mapping using the UCSC Genome Institute genome browser. We used the genome of Canis lupus familaris Boxer breed [GCF_000002285.3, (CanFam 3.1)] as a reference to compare the gene fragments and SNPs. We associated SNPs with the CMC and control groups by testing odds ratios (OR) through Fisher’s exact tests to determine an association or risk for CMC. We detected two SNPs for ESR1, three for MUC1, six for HER2, and one for BRCA1. MUC1 was the only gene to display an SNP in an exonic region that resulted in an amino acid substitution (Pro>Thr). No significant differences based on the OR were found, though the majority of SNPs, with the exception of four, were found in females with CMC. We report a novel molecular marker for HER2 that amplifies exons 25–26 and introns 24-25, and highlight the importance of conducting further studies on MUC1 and elucidating the role of introns and splicing in candidate genes associated with CMC.Diljem svijeta, tumor mliječne žlijezde kuja (CMC) najčešća je skupina neoplazija kuja te je tri puta učestalija u pasa nego u ljudi. U Kolumbiji, CMC je bio druga najčešća vrsta tumora, nakon neoplazija na koži. Nadalje, genetika je jedan od najvažnijih čimbenika uključenih u bilo koju vrstu tumora, a genetska baza ove bolesti održava se kroz linijski uzgoj uslijed promjena alelnih frekvencija, koje se mogu pratiti preko molekularnih markera. Ova studija imala je za cilj detektirati polimorfizme jednog nukleotida (SNP-e) povezane s CMC-om u uzorcima krvi prikupljenih od kuja s CMC-om i zdravih kuja u veterinarskoj bolnici Diego Villegas Toro Sveučilišta Caldas (Manizales, Kolumbija). Dizajnirali smo početnice uporabom Primer-BLAST i Primer3 te su fragmenti gena HER2, MUC1, ESR1 i BRCA1 pojačani za identifikaciju SNP-a preko mapiranja genoma uporabom preglednika genoma Instituta za genom Sveučilišta Santa Cruz u Kaliforniji (UCSC). Rabili smo genom Canis lupus familaris pasmine bokser [GCF_000002285.3, (CanFam 3.1)] kao referencu za usporedbu fragmenata gena i SNP-a. Povezali smo SNP-e s CMC-om i kontrolnim skupinama testiranjem omjera izgleda (OR) pomoću Fisherovih egzaktnih testova za određivanje povezanosti ili rizika od CMC-a. Detektirali smo dva SNP-a za ESR1, tri za MUC1, šest za HER2 i jedan za BRCA1. MUC1 je bio jedini gen koji je pokazao SNP u regiji egzona što je rezultiralo supstitucijom aminokiseline (Pro>Thr). Nismo pronašli značajne razlike na temelju OR-a, premda je većina SNP-a, izuzev četiri, pronađena u kuja s CMC-om. Prijavljujemo novi molekularni marker za HER2 koji pojačava egzone 25 - 26 i introne 24 - 25 te naglašavamo važnost provođenja dodatnih studija na MUC1, kao i pojašnjenja uloge introna i izrezivanja u gena kandidata poveznih s CMC-om

Procjena gama-aktina, beta-aktina, gliceraldehid-3-fosfat dehidrogenaze i 18S kao referentnih gena za qRT-PCR uporabom uzoraka krvi u istraživanju mliječnih žlijezda kujica

Mammary tumours are the most frequent group of neoplasia in female dogs. Tumorigenesis is associated with gene expression changes in a wide variety of genes. For this reason, real-time quantitative PCR (qRT-PCR) is used in routine diagnostic procedures in clinical practice due to its the specificity, sensitivity, simplicity, and high performance. qRT-PCR is also widely used to measure the expression of target genes compared to reference genes in several tissues. We collected blood samples from healthy female dogs and females with canine mammary cancer in Manizales, Colombia between June 2018 and January 2019, and mRNA was isolated from each sample for cDNA synthesis. qRT-PCR-based expression assays were performed using primers designed for gamma-actin, beta-actin, GAPDH, and 18S genes. We calculated the amplification efficiency, specificity, and stability using geNorm, NormFinder, BestKeeper, and the ΔCt comparative method. We obtained linear regressions to verify constant gene expression and conducted an ANOVA to detect expression differences regarding Ct values and healthy vs. ill conditions. We found stability for primers 18S-1, GAPDH-1, GAPDH-NM, and Gamma-actin-1 (in increasing order). Furthermore, these genes showed constant expression levels in patients (R2>0.80). We report novel primers for gamma-actin and GAPDH, which proved to be efficient endogenous control genes for qRT-PCR applications in blood tissue. These primers are useful for gene expression research in canine mammary cancer.Tumori mliječnih žlijezda najčešća su skupina neoplazija u kujica. Tumorogeneza je povezana s promjenama u ekspresiji gena u širokom rasponu gena. Iz tog razloga se rabi kvantitativna lančana reakcija polimerazom u stvarnom vremenu (qRT-PCR) u rutinskim dijagnostičkim postupcima u kliničkoj praksi, uslijed specifičnosti, osjetljivosti, jednostavnosti i visoke učinkovitosti ove tehnike. qRT-PCR se široko rabi i za mjerenje ekspresije ciljanih gena u usporedbi s referentnim genima u više vrsta tkiva. Prikupili smo uzorke krvi zdravih kujica i kujica s tumorom mliječnih žlijezda u Manizalesu, Kolumbiji, od lipnja 2018. do siječnja 2019. godine. Izolirali smo mRNK iz svakog uzorka za cDNK sintezu. Pokusi ekspresije na bazi qRT-PCR obavljeni su uporabom primera dizajniranih za gama-aktin, beta-aktin, gliceraldehid- 3-fosfat dehidrogenazu (GAPDH) i 18S gene. Izračunali smo pojačanu učinkovitost, specifičnost i stabilnost uporabom geNorm, NormFinder, BestKeeper i ΔCt komparativne metode. Dobili smo linearne regresije za potvrđivanje stalne ekspresije gena i proveli smo analizu varijance (ANOVA) za detekciju razlika u ekspresiji s obzirom na Ct vrijednosti te zdrava u usporedbi s bolesnim stanjima. Otkrili smo i stabilnost za primere 18S-1, GAPDH-1, GAPDH-NM i Gamma-actin-1 (rastućim redoslijedom). Nadalje, ovi geni su pokazali konstantne razine ekspresije u pacijenata (R2>0,80). Izvještavamo o novim primerima za gama-aktin i GAPDH, koji su se pokazali učinkovitim endogenim kontrolnim genima za qRT-PCR primjene u krvnom tkivu. Ti primeri su korisni za istraživanje ekspresije gena u tumora psećih mliječnih žlijezda

Pilot multicenter study to determine the utility of point-of-care ultrasound to predict difficulty of tracheal intubation using videolaryngoscopy with the McGrath™ Mac videolaryngoscope.

BACKGROUND Clinical airway screening tests used to predict difficulties during airway management have low sensitivity and specificity. Point-of-care airway ultrasound has described measurements related to problems with difficult direct laryngoscopy. Nevertheless, the correlation between ultrasound parameters and videolaryngoscopy has not been published yet. The aim of this multicenter, prospective observational pilot study was to evaluate the applicability of clinical parameters and ultrasound measurements to find potential tracheal intubation difficulties when videolaryngoscopy is used. METHODS Preoperatively, six clinical airway assessments were performed: (1) modified Mallampati score, (2) thyromental distance, (3) sternomental distance, (4) interincisal distance, (5) upper lip bite test, and (6) neck circumference. Six ultrasound parameters were measured in awake patients: (1) distance from skin to hyoid bone, (2) distance from skin to epiglottis, (3) hyomental distance in neutral head position, (4) hyomental distance in head-extended position, (5) distance from skin to the deepest part of the palate, and (6) sagittal tongue area. And finally, there was one ultrasound measure obtained in anesthetized patients, the compressed sagittal tongue area during videolaryngoscopy. The difficulty for tracheal intubation using a McGrath™ Mac videolaryngoscope, the percentage of glottic opening, and Cormack-Lehane grade were also assessed. RESULTS In this cohort of 119 subjects, tongue dimensions, particularly the sagittal tongue area, showed a robust association with increased intubation difficulty using videolaryngoscopy. A multiparametric model combining the following three ultrasound variables in awake patients: (a) the distance from skin to epiglottis, (b) the distance from skin to the deepest part of the palate, and (c) the sagittal tongue area, yielded a sensitivity of 92.3%, specificity of 94.5%, positive predictive value of 82.8%, and negative predictive value of 97.8% (p < 0.001). CONCLUSION Point-of-care airway ultrasound emerges as a more useful tool compared to traditional clinical scales to anticipate possible challenges during videolaryngoscopic intubation

Herpes Simplex Virus Type 1 Infection of the Central Nervous System: Insights Into Proposed Interrelationships With Neurodegenerative Disorders

Herpes simplex virus type 1 (HSV-1) is highly prevalent in humans and can reach the brain without evident clinical symptoms. Once in the central nervous system (CNS), the virus can either reside in a quiescent latent state in this tissue, or eventually actively lead to severe acute necrotizing encephalitis, which is characterized by exacerbated neuroinflammation and prolonged neuroimmune activation producing a life-threatening disease. Although HSV-1 encephalitis can be treated with antivirals that limit virus replication, neurological sequelae are common and the virus will nevertheless remain for life in the neural tissue. Importantly, there is accumulating evidence that suggests that HSV-1 infection of the brain both, in symptomatic and asymptomatic individuals could lead to neuronal damage and eventually, neurodegenerative disorders. Here, we review and discuss acute and chronic infection of particular brain regions by HSV-1 and how this may affect neuron and cognitive functions in the host. We review potential cellular and molecular mechanisms leading to neurodegeneration, such as protein aggregation, dysregulation of autophagy, oxidative cell damage and apoptosis, among others. Furthermore, we discuss the impact of HSV-1 infection on brain inflammation and its potential relationship with neurodegenerative diseases

Therapeutic approach with commercial supplements for pantothenate kinase-associated neurodegeneration with residual PANK2 expression levels

[Background]: Neurodegeneration with brain iron accumulation (NBIA) is a group of rare neurogenetic disorders frequently associated with iron accumulation in the basal nuclei of the brain characterized by progressive spasticity, dystonia, muscle rigidity, neuropsychiatric symptoms, and retinal degeneration or optic nerve atrophy. Pantothenate kinase-associated neurodegeneration (PKAN) is one of the most widespread NBIA subtypes. It is caused by mutations in the gene of pantothenate kinase 2 (PANK2) that result in dysfunction in PANK2 enzyme activity, with consequent deficiency of coenzyme A (CoA) biosynthesis, as well as low levels of essential metabolic intermediates such as 4′-phosphopantetheine, a necessary cofactor for essential cytosolic and mitochondrial proteins. [Methods]: In this manuscript, we examined the therapeutic effectiveness of pantothenate, panthetine, antioxidants (vitamin E and omega 3) and mitochondrial function boosting supplements (L-carnitine and thiamine) in mutant PANK2 cells with residual expression levels. [Results]: Commercial supplements, pantothenate, pantethine, vitamin E, omega 3, carnitine and thiamine were able to eliminate iron accumulation, increase PANK2, mtACP, and NFS1 expression levels and improve pathological alterations in mutant cells with residual PANK2 expression levels. [Conclusion]: Our results suggest that several commercial compounds are indeed able to significantly correct the mutant phenotype in cellular models of PKAN. These compounds alone or in combinations are of common use in clinical practice and may be useful for the treatment of PKAN patients with residual enzyme expression levels.This work was supported by FIS PI16/00786 and PI19/00377 grants, Instituto de Salud Carlos III, Spain and Fondo Europeo de Desarrollo Regional (FEDER-Unión Europea), Proyectos de Investigación de Excelencia de la Junta de Andalucía CTS-5725 and PY18-850