Bacteriological resolve of pyoderma associated with canine demodicosis without antibiotic/antiseptic therapy

Generalized canine demodicosis is a disease caused by Demodex spp. commonly associated with infection by Staphylococcus spp., normal inhabitants of dogs skin. Both microorganisms proliferate within the hair follicles causing folliculitis and furunculosis. In the past, systemic antibiotic therapy was supported for all dogs with secondary bacterial infection. Nowadays, as the incidence of skin infections with multi-resistant bacteria is increasing, a judicious use of systemic antibiotics is recommended.Trabajo publicado en Cagliada, Maria del Pilar Lilia y Galosi, Cecilia Mónica (comps.). I Congreso de Microbiología Veterinaria. Libro de resúmenes. La Plata: Facultad de Ciencias Veterinarias, 2021.Facultad de Ciencias Veterinaria

Whole genome sequencing and de novo assembly of Staphylococcus pseudintermedius: a pangenome approach to unravelling pathogenesis of canine pyoderma

Background Staphylococcus pseudintermedius is the main aetiological agent of canine pyoderma. Whole genome sequencing is the most comprehensive way of obtaining relevant genomic information about micro-organisms. Hypothesis/Objectives Oxford Nanopore technology enables quality sequencing and de novo assembly of the whole genome of S. pseudintermedius. Whole genome analysis of S. pseudintermedius may help to better understand the pathogenesis of canine pyodermas. Methods and materials Twenty-two strains of S. pseudintermedius isolated from the skin of five healthy dogs and 33 strains isolated from skin of 33 dogs with pyoderma were analysed. DNA was extracted and sequenced using Oxford Nanopore MinION, a new technology that delivers longer reads in a hand-held device. The pangenome was analysed and visualised with Anvi’o 6.1. Results Nanopore technology allowed the sequencing and de novo assembly of the genomes of 55 S. pseudintermedius strains isolated from healthy dogs and from dogs with pyoderma. The average genome size of S. pseudintermedius was 2.62 Mbp, with 48% being core genome. Pyoderma isolates contained a higher number of antimicrobial resistance genes, yet the total number of virulence factors genes did not change between isolates from healthy dogs and from dogs with pyoderma. Genomes of meticillin-resistant S. pseudintermedius (MRSP) strains were larger than those of meticillin-susceptible (MSSP) strains (2.80 Mbp versus 2.59 Mbp), as a consequence of a greater presence of antimicrobial resistance genes, phages and prophages. Conclusions and clinical importance This technique allows much more precise and easier characterisation of canine S. pseudintermedius populations and may lead to a better understanding of the pathogenesis of canine pyodermas.info:eu-repo/semantics/publishedVersio

Whole genome sequencing and de novo assembly of Staphylococcus pseudintermedius: a pangenome approach to unravelling pathogenesis of canine pyoderma

Background: Staphylococcus pseudintermedius is the main aetiological agent of canine pyoderma. Whole genome sequencing is the most comprehensive way of obtaining relevant genomic information about micro-organisms. Hypothesis/Objectives: Oxford Nanopore technology enables quality sequencing and de novo assembly of the whole genome of S. pseudintermedius. Whole genome analysis of S. pseudintermedius may help to better understand the pathogenesis of canine pyodermas. Methods and materials: Twenty-two strains of S. pseudintermedius isolated from the skin of five healthy dogs and 33 strains isolated from skin of 33 dogs with pyoderma were analysed. DNA was extracted and sequenced using Oxford Nanopore MinION, a new technology that delivers longer reads in a hand-held device. The pangenome was analysed and visualised with Anvi’o 6.1. Results: Nanopore technology allowed the sequencing and de novo assembly of the genomes of 55 S. pseudintermedius strains isolated from healthy dogs and from dogs with pyoderma. The average genome size of S. pseudintermedius was 2.62 Mbp, with 48% being core genome. Pyoderma isolates contained a higher number of antimicrobial resistance genes, yet the total number of virulence factors genes did not change between isolates from healthy dogs and from dogs with pyoderma. Genomes of meticillin-resistant S. pseudintermedius (MRSP) strains were larger than those of meticillin-susceptible (MSSP) strains (2.80 Mbp versus 2.59 Mbp), as a consequence of a greater presence of antimicrobial resistance genes, phages and prophages. Conclusions and clinical importance: This technique allows much more precise and easier characterisation of canine S. pseudintermedius populations and may lead to a better understanding of the pathogenesis of canine pyodermas.Fil: Ferrer, Lluís. Universitat Autònoma de Barcelona; EspañaFil: García Fonticoba, Rocío. Universitat Autònoma de Barcelona; EspañaFil: Pérez, Daniel. Universitat Autònoma de Barcelona; EspañaFil: Viñes, Joaquim. Universitat Autònoma de Barcelona; EspañaFil: Fàbregas, Norma. Universitat Autònoma de Barcelona; EspañaFil: Madroñero, Sergi. Universitat Autònoma de Barcelona; EspañaFil: Meroni, Gabriele. No especifíca;Fil: Martino, Piera A.. No especifíca;Fil: Martínez, Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Maté, María Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Sanchez Bruni, Sergio Fabian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Cuscó, Anna. Universitat Autònoma de Barcelona; EspañaFil: Migura García, Lourdes. Universitat Autònoma de Barcelona; EspañaFil: Francino, Olga. Universitat Autònoma de Barcelona; Españ

Blockade of Stat3 oncogene addiction induces cellular senescence and reveals a cell-nonautonomous activity suitable for cancer immunotherapy

Stat3 is constitutively activated in several tumor types and plays an essential role in maintaining their malignant phenotype and immunosupression. To take advantage of the promising antitumor activity of Stat3 targeting, it is vital to understand the mechanism by which Stat3 regulates both cell autonomous and non-autonomous processes. Here, we demonstrated that turning off Stat3 constitutive activation in different cancer cell types induces senescence, thus revealing their Stat3 addiction. Taking advantage of the senescence-associated secretory phenotype (SASP) induced by Stat3 silencing (SASP-siStat3), we designed an immunotherapy. The administration of SASP-siStat3 immunotherapy induced a strong inhibition of triplenegative breast cancer and melanoma growth associated with activation of CD4 + T and NK cells. Combining this immunotherapy with anti-PD-1 antibody resulted in survival improvement in mice bearing melanoma. The characterization of the SASP components revealed that type I IFN-related mediators, triggered by the activation of the cyclic GMP-AMP synthase DNA sensing pathway, are important for its immunosurveillance activity. Overall, our findings provided evidence that administration of SASP-siStat3 or low dose of Stat3- blocking agents would benefit patients with Stat3-addicted tumors to unleash an antitumor immune response and to improve the effectiveness of immune checkpoint inhibitors.Fil: de Martino, Mara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Tkach, Mercedes. Institute Curie; FranciaFil: Bruni, Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Rocha, Darío Gastón. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales; ArgentinaFil: Mercogliano, María Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Cenciarini, Mauro Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Chervo, María Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Proietti Anastasi, Cecilia Jazmín. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Dingli, Florent. Institute Curie; FranciaFil: Loewy, Ruth Miriam. Institute Curie; FranciaFil: Fernández, Elmer A.. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas. Universidad Católica de Córdoba. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas; ArgentinaFil: Elizalde, Patricia Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Piaggio, Eliane. Institute Curie; FranciaFil: Schillaci, Roxana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentin

Emerging Targeted Therapies for HER2-Positive Breast Cancer

Breast cancer is the most common cancer in women and the leading cause of death. HER2 overexpression is found in approximately 20% of breast cancers and is associated with a poor prognosis and a shorter overall survival. Tratuzumab, a monoclonal antibody directed against the HER2 receptor, is the standard of care treatment. However, a third of the patients do not respond to therapy. Given the high rate of resistance, other HER2-targeted strategies have been developed, including monoclonal antibodies such as pertuzumab and margetuximab, trastuzumab-based antibody drug conjugates such as trastuzumab-emtansine (T-DM1) and trastuzumab-deruxtecan (T-DXd), and tyrosine kinase inhibitors like lapatinib and tucatinib, among others. Moreover, T-DXd has proven to be of use in the HER2-low subtype, which suggests that other HER2-targeted therapies could be successful in this recently defined new breast cancer subclassification. When patients progress to multiple strategies, there are several HER2-targeted therapies available; however, treatment options are limited, and the potential combination with other drugs, immune checkpoint inhibitors, CAR-T cells, CAR-NK, CAR-M, and vaccines is an interesting and appealing field that is still in development. In this review, we will discuss the highlights and pitfalls of the different HER2-targeted therapies and potential combinations to overcome metastatic disease and resistance to therapy

Development of a qPCR to evaluate accurately the treatment response of dogs with demodicosis

Canine demodicosis is the result of an overpopulation of Demodex mites, normal inhabitants of most skin of mammals. There is no reliable method to quantify the number of Demodex mites present in the skin to provide accurate information about treatment response. The objective of this study was to develop a molecular technique to quantify the Demodex load in canine skin in order to evaluate the conventional treatment response in dogs with demodicosis. For this purpose, a real time qPCR was developed using primers that amplified a fragment of the Demodex canis 18S rRNA gene. Ten-fold dilutions of the DNA extracted from isolated mites were used to elaborate the standard curve. In addition, 44 skin biopsies (4 mm) obtained from nine canine cadavers and 18 skin biopsies from seven dogs with demodicosis in days zero and 14, 35 and 56 post-treatment (Animal Welfare Committee, FCV-UNCPBA) were used to assess the number of mites in the skin of healthy dogs and of dogs with demodicosis. The skin biopsies of healthy dogs were negative in 38 samples, being positive (>one mite) the remaining samples (six). However, in skin biopsies of diseased dogs the initial Demodex load was mostly >100 mites, decreasing to 50-100, 10-50 and 0-10 mites at 14, 35 and 56 days post-treatment, respectively. The homology between the qPCR product and the 18S rRNA gene was confirmed by DNA sequencing (GenBank KC010485 - Demodex canis). This simple technique could be a useful tool to evaluate accurately the response to treatment of dogs with demodicosis.Fil: Martínez, Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Sastre, Natalia. Universidad Autonoma de Barcelona. Facultad de Veterinaria; EspañaFil: Francino, Olga. Universidad Autonoma de Barcelona. Facultad de Veterinaria; EspañaFil: Maté, María Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Sanchez Bruni, Sergio Fabian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: del Sole, Maria Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Ferrer, Lluis. Universidad Autonoma de Barcelona. Facultad de Veterinaria; España9th World Congress Of Veterinary DermatologyAustraliaWorld Association of Veterinary Dermatolog

Clinical, cytologic and microbiological evidence of pyoderma recovery in dogs with demodicosis without antimicrobial therapy

Generalized pustular canine demodicosis is the result of an overpopulation of mites of Demodex spp. commonly associated with infection by Staphylococcus spp., normal inhabitants of the skin of most mammals. Both microorganisms proliferate within the hair follicles causing follicular hyperkeratosis in the head, trunk and legs. Dermatological examination reveals signs such as erythema, alopecia, folliculitis and furunculosis. The diagnosis is confirmed by skin scraping and cytology of the lesions, microscopically identifying mites and inflammatory cells with cocci, respectively. In the past, systemic antibiotic therapy was supported for all dogs with secondary bacterial infection. Nowadays, as the incidence of skin infections with multi-resistant bacteria is increasing, a judicious use of systemic antibiotics is recommended. Thus, the Clinical Practice Guidelines on Demodicosis Treatment (2020) recommend topical antibiotic therapy in dogs with generalized demodicosis up to 1-2 weeks after clinical and cytologic resolution of the skin infection. The main goal of this study was to assess the clinical-cytological evolution and bacteriological cure of canines with generalized demodicosis treated only with acaricides. For this purpose, 7 (seven) patients with pustular demodicosis were admitted for their attention at the Teaching Hospital of Small Animals (HEPA). On day 0, the animals were clinically evaluated and clinical score awarded; cytological test and skin swabs were obtained from lesions. Then, patients initiated a non-antibiotic treatment as follow: 4 (four) dogs were treated with afoxolaner at a dose of 2.7-6.9 mg/kg on day 0 and day 28, and 3 (three) dogs were treated with oral ivermectin at a dose of 0.5 mg/kg/24 h for 63 days. On days 14, 35 and 56 post-treatment, clinical score and cytological results were recorded and sample swabs from skin lesions were obtained. Cytology samples were stained with Giemsa Merck® by conventional technique and observed microscopically by the same dermatologist. Sample swabs were stored in Stuart medium up to overnight growth on Tryptic Soy Agar medium supplemented with 10% sterile bovine blood and the phenotypic identification bacterial strains were performed by conventional biochemical techniques. Clinical score (mean +/- DE) decreased considerably throughout the treatment as follows: 31.8 +/- 3.4, 28.0 +/- 5.4, 13.2 +/- 5.0, and 4.8 +/- 1.5 on day 0, 14, 35 and 56, respectively. Cytology with pyoderma characteristics were positive at days 0 and 14 post-treatment in all dogs, and became negative in 5 dogs at day 35, being negative in all dogs (6) at day 56 . Staphylococcus spp. were isolated from skin samples in all dogs on days 0 and became negative in 6 dogs at day 56 post-treatment. In conclusion, these preliminary results propose that pyoderma associated with canine demodicosis could resolve clinical, cytological and bacteriologically with single acaricidal therapy, avoiding systemic or topical antibiotic therapy.Fil: Martínez, Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Cacciato, Claudio Santiago. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; ArgentinaFil: Fogel, Fernando Adrián. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Departamento de Clinicas. Hospital Escuela; ArgentinaFil: Maté, María Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Sanchez Bruni, Sergio Fabian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: del Sole, Maria Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina1ras. Jornadas del Instituto de Investigaciones Clínicas VeterinariasArgentinaUniversidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigaciones Clínicas Veterinaria